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Biomedical subjects

A M Edwards

Publications and source records attributed to A M Edwards.

At least 145 records · Page 8Linked to original sources
PubMed1985

Effects of hemin on rat liver cyclic AMP-dependent protein kinases in cell extracts and intact hepatocytes.

Cyclic AMP-dependent protein kinases I and II, partially purified from rat liver cytosol, were inhibited 50% by 40 microM hemin and 100 microM hemin, respectively. With the purified catalytic subunit of cyclic AMP-dependent protein kinase, hemin caused non-competitive inhibition with respect to the peptide substrate and mixed inhibition with respect to ATP. Hemin also inhibited purified phosphorylase b kinase, indicating that hemin concentrations above 10 microM markedly inhibit multiple protein kinases. In isolated intact hepatocytes, hemin inhibited the glucagon-dependent activation of cyclic AMP-dependent protein kinases and the activation of glycogen phosphorylase. For both effects, high heme concentrations (40-60 microM) were required for 50% inhibition. Similar high levels of exogenous hemin inhibited total hepatocyte protein synthesis. By contrast, 5 microM hemin or less was sufficient to raise intracellular heme levels, as indicated by the relative heme-saturation of tryptophan oxygenase in hepatocytes. Hemin, 5 microM, completely repressed induction of 5-aminolevulinate synthase by dexamethasone in hepatocyte primary cultures. Such repression is unlikely to be mediated by inhibition of protein kinases.

5-Aminolevulinate Synthetase↗
PubMed1985

Phenobarbital and some other liver tumor promoters stimulate DNA synthesis in cultured rat hepatocytes.

Hepatocytes isolated from normal adult rats were maintained at sub-confluent density in a defined medium in primary culture. In control cultures with added epidermal growth factor and low concentrations of dexamethasone, the rate of DNA synthesis was initially low but increased after about 30 hours in culture. Addition of the xenobiotic liver tumor promoters phenobarbital, alpha-hexachlorocyclohexane or p,p'-dichlorodiphenyltrichloroethane to cultures after cell attachment caused concentration-dependent stimulation of DNA synthesis measured after 2 days in culture. While dexamethasone (30 nM) alone had little effect on hepatocyte DNA synthesis, the stimulatory effects of the xenobiotics required the permissive presence of the steroid.

Animals↗
PubMed1985

Photochemical reactivity of the homologous proteins alpha-lactalbumin and lysozyme.

The fluorescent behaviour and the photodynamic effect was studied in native and structurally modified lysozyme and alpha-lactalbumin. The Tyr residues in lysozyme and alpha-lactalbumin show different sensitivities to the photodynamic effect. The effect is zero in the case of Tyr from native lysozyme. In contrast, the Tyr residues in alpha-lactalbumin are susceptible to photooxidation, which indicates a greater degree of exposure to the solvent. The three His residues of alpha-lactalbumin have different degrees of exposure and show two different kinetics of photooxidation whereas the His residue of lysozyme is photooxidized with a single kinetic. Two photooxidation kinetics were obtained for the Trp residues of both native proteins, an indication that in both cases there are Trp residues that are differently exposed to the solvent. The wavelengths of maximum fluorescent emission of the Trp residues were different for the two proteins, an effect which can also be explained in terms of a difference in the environment of these residues. The modified form of these proteins emit at wavelengths longer than those of the native forms. When modified the proteins photooxidize with noticeably greater quantum yields.

Lactalbumin↗
PubMed1985

The characterization and cellular distribution of a family of antigens related to myelin associated glycoprotein in the developing nervous system.

The antigenic epitope detected on myelin associated glycoprotein (MAG) by the monoclonal antibody HNK-1 (Leu 7) was sensitive to degradation by trifluoromethane-sulfonic acid (TFMS) and is therefore probably carbohydrate in nature. This antigen was found to be widely distributed within the rat and chicken embryonic nervous system and was present on cultured central and peripheral neurons (100%), oligodendrocytes (100%) and astrocytes (70-80%) as detected by double marker immunofluorescence. The antigen could be removed from cultured neurons by trypsinization and its resynthesis was blocked by cycloheximide, suggesting that the carbohydrate epitope detected by HNK-1 was attached to a de novo synthesized protein. Several molecular species were detected on Western blots of detergent extracts from 13-15d rat embryonic brain and neuron-enriched cultures from chick spinal cord and dorsal root ganglia. Protein components with molecular weights in the ranges of 90-100 kd to 280 kd were observed and comprise a family of glycoproteins containing the HNK-1 reactive carbohydrate epitope present on MAG. These glycoproteins could play a role in intercellular interactions within the developing nervous system.

Animals↗
PubMed1985

Induction of gamma-glutamyl transpeptidase in primary cultures of normal rat hepatocytes by liver tumor promoters and structurally related compounds.

Rat hepatocytes maintained for up to 6 days in primary culture were used to test a variety of xenobiotics and steroids for effects on the activity of gamma-glutamyltranspeptidase (GGT) in normal cells. In control cultures GGT activity was low and increased slowly with time. When added to cultures for 5 days, a variety of xenobiotics and steroids increased GGT activity to levels 2- to 6-times those of control cultures. Induction of GGT was potentiated for most test compounds by 20-30 nM dexamethasone and diminished by nicotinamide or adenosine-3',5'-monophosphate. Effective non-genotoxic inducers included phenobarbital and some structurally related compounds, p,p'-dichlorodiphenyltrichloroethane,alpha- and gamma-hexachlorocyclohexanes, Aroclor 1254, butyl hydroxytoluene, nafenopin, various estrogens, progesterone, pregnenolone-16 alpha-carbonitrile and cyproterone acetate. A number of compounds including barbituric acid, butyl hydroxyanisole, acetaminophen, saccharin, caffeine, clofibrate and some bile acids failed to induce GGT. Except for 2-acetylaminofluorene and diethylnitrosamine, genotoxic compounds tested did not increase GGT. The results establish that a structurally diverse group of xenobiotics and steroids, many of which are considered to be liver tumour promoters, may directly enhance GGT gene expression in normal hepatocytes. Thus, a variety of compounds used in experimental studies of liver cancer induction as promoters may elevate GGT by mechanism(s) not necessarily related to carcinogenesis.

Animals↗
PubMed1984

Relationship between inductions of monooxygenase activity and gamma-glutamyltranspeptidase in rat hepatocyte primary cultures.

The proposition that changes in activity of gamma-glutamyltranspeptidase (GGT) in serum may provide a useful index of the extent of induction of liver drug-metabolizing enzymes by various drugs was examined by comparing control of GGT and monooxygenase activities in cultured hepatocytes. In rat hepatocyte monolayers maintained for up to 5 days the effects of xenobiotics and other factors on cellular GGT activity were compared with effects on a relatively broad measure of drug metabolism, the 7-ethoxycoumarin O-deethylase (ECD) activity of intact cells. A diverse group of drugs including phenobarbital and other barbiturates, diphenylhydantoin, glutethimide, aminopyrine and griseofulvin and the steroids dexamethasone and pregnenolone 16 alpha-carbonitrile were shown to induce both GGT and ECD under comparable culture conditions. Inductions of both activities were potentiated by glucocorticoids and depressed (where tested) by dibutyryl cyclic AMP. Some other hormones or nutrients modulated the activities differently. The magnitude of GGT induction by different drugs did not correlate with relative ECD induction and for several drugs the concentration-dependence of the two effects was different. Interpretation is complicated by the possible contribution of multiple forms of cytochrome P-450 to ECD activity but it seems unlikely that drugs which induce both GGT and drug metabolism do so via a common regulatory mechanism. For such drugs changes in serum GGT could provide only a crude guide to likely changes in drug metabolism. Some compounds including polycyclic hydrocarbons and warfarin induced ECD but had no associated effect on GGT in hepatocytes.

Animals↗
PubMed1984

7-Ethoxycoumarin deethylase activity as a convenient measure of liver drug metabolizing enzymes: regulation in cultured rat hepatocytes.

Assays of 7-ethoxycoumarin O-deethylase (ECD) activity in intact cells were used as a sensitive and convenient measure of the drug-metabolizing activity of rat hepatocytes maintained for up to 4 days in primary culture. A combination of nicotinamide or other pyridines with dexamethasone was shown to maintain ECD at or above the activity of untreated livers in vivo and to potentiate induction by xenobiotics. Inductions in vivo and in culture were quantitatively similar but differed qualitatively as judged by the proportion of ECD activity inhibitable by metyrapone. A survey of possible endogenous regulators of liver monooxygenases established that: dexamethasone and other glucocorticoids induced ECD and potentiated induction by xenobiotics, particularly phenobarbitone; other steroids including testosterone, 17 beta-estradiol and pregnenolone 16 alpha-carbonitrile caused small inductions; insulin lowered both ECD activity and the proportion of activity inhibitable by metyrapone; dibutyryl cyclic AMP or glucagon lowered ECD; and high concentrations of aminolevulinate partly repressed induction by xenobiotics. Based on these findings, hepatocyte culture conditions which maintain ECD activity and inducibility at or above in vivo levels are defined.

7-Alkoxycoumarin O-Dealkylase↗
PubMed1984

Molecular characteristics of the epitope in myelin-associated glycoprotein that is recognized by a monoclonal IgM in human neuropathy patients.

The antigen for the IgM monoclonal antibody from patients with IgM paraproteinemia and peripheral neuropathy is the myelin-associated glycoprotein (MAG), a minor protein component of both human PNS myelin and human CNS myelin. Sera from five patients were found to react with identical proteolytically derived fragments of MAG indicating that the monoclonal IgM from these patients is recognizing a common epitope. Furthermore, the lectin concanavalin A reacts with these fragments and deglycosylation of isolated MAG abolishes the recognition of MAG by the patient monoclonal IgM. Therefore, it appears that the monoclonal IgM from these five patients recognizes a common epitope which contains carbohydrate moieties. These data are consistent with the idea that the peripheral myelin sheath is involved in an autoimmune response directed against MAG.

Blood Proteins↗
PubMed1982

Regulation of gamma-glutamyltranspeptidase in rat hepatocyte monolayer cultures.

Hepatocytes isolated as a relatively pure population from normal adult rats were maintained in primary monolayer culture for 4 to 7 days on plastic dishes. Factors affecting activity of gamma-glutamyltranspeptidase (GGT) in culture were investigated to establish a basis for in vitro studies on carcinogen-induced changes in GGT. Freshly plated cultures contained few GGT-positive cells (0.4%) and very low total GGT activity, but with all media tested, activity increased progressively with time in culture, the extent of increase depending on medium composition. For hepatocytes in modified Waymouth medium alone (control cultures), there was a slow rise in activity after a 1- to 2-day lag period. This increase was enhanced up to 10-fold in the presence of 3 microM dexamethasone for cells on plastic but only 2- to 3-fold for cells on collagen gels. The time- and dose-dependent action of dexamethasone was prevented by cycloheximide, partly blocked by actinomycin D, and reversed after several hr by removing the steroid, when GGT activity decreased with a maximum half-life of 60 to 80 hr. These observations suggest that dexamethasone caused reversible enzyme induction dependent on continuing RNA and protein synthesis. Cell maintenance with fetal calf serum or with N6,O2'-dibutyryl adenosine 3':5'-monophosphate or glucagon markedly reduced the extent of induction by dexamethasone and slightly lowered activity in control cultures. Maintenance at lower pH within the range of 7.2 to 7.8 or at higher glucose concentrations within the range of 2.8 to 28 mM also resulted in markedly lower GGT activities in control or dexamethasone-induced cultures. Glucose repression was independent on insulin concentration. The reversible induction and repression of GGT may reflect broad changes in hepatocyte gene expression rather than specific controls of GGT function. Since GGT in cultured hepatocytes is subject to regulation by a variety of noncarcinogens, caution is required in its use as a preneoplastic marker. Nevertheless, the identification of culture media which preserve low activity may allow studies on carcinogen-induced changes in GGT as a probe of the early events in vitro hepatocarcinogenesis.

Animals↗
PubMed1980

Chemo-immunotherapy with levamisole in metastatic breast carcinoma: a controlled clinical trial.

A randomized prospectively controlled trial assessing the value of levamisole in patients receiving chemotherapy for metastatic carcinoma of the breast is reported. Patients randomized to the chemo-immunotherapy group received levamisole 2.5 mg/kg orally for 2 consecutive days each week while off chemotherapy, which consisted of the standard regiment of cyclophosphamide, methotrexate and 5-fluorouracil (CMF). Seventy-seven patients were assessable, 38 patients in the chemo-immunotherapy arm and 39 patients in the chemotherapy alone arm. Of the 38 patients in the chemo-immunotherapy, 18 (47.3%) experienced a complete or partial response compared to 18 (46.1%) in the chemotherapy alone group. The median duration of response in both groups of patients was approximately 15 months. The median duration of survival in the chemo-immunotherapy group was 21 months and in the chemotherapy alone group has not yet been reached but is at least 21 months. Levamisole was generally well tolerated, but in the dosage and scheduling did not appear to improve the results of the chemotherapy. It is suggested that efforts should be directed to increasing the complete response rate with chemotherapy and radiotherapy to provide the best setting for immunotherapy trials.

Antineoplastic Agents↗
PubMed1979

Measurement of cholic acid synthesis and secretion by isolated rat hepatocytes.

Liver cells isolated from normal and cholestyramine-treated rats were incubated as cell suspensions for up to 4 hr in a simple, defined medium. The bile acid concentration in cells plus cell medium was determined by gas-liquid chromatography. Normal hepatocytes synthesized cholic acid at an initial rate of 0.25 nmol/mg cell protein per hr, which is comparable to rates reported from in vivo methods. This rate was increased more than 4-fold when rats were fed a cholestyramine-containing diet for 7 days prior to liver cell isolation. Although cholic acid was secreted into the cell medium during the incubation, it could not be assayed reliably by the hydroxysteroid dehydrogenase assay method, contrary to the reports of Anwer et al. 1975. Biochem. Biophys. Res. Commun. 64: 603 and Gardner and Chenouda 1978. J. Lipid Res. 19: 985.

Animals↗
PubMed1978

Controlled trial of disodium cromoglycate in chronic persistent ulcerative colitis.

Oral disodium cromoglycate (200 mg qds) has been tested in 26 patients with ulcerative colitis that was resistant to medical treatment. In a double-blind crossover trial disodium cromoglycate and placebo were added to conventional treatment in random order, each for four weeks. There was no significant difference in therapeutic effect between disodium cromoglycate and placebo.

Adolescent↗