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Biomedical subjects

M J Wilson

Publications and source records attributed to M J Wilson.

At least 181 records · Page 10Linked to original sources

Human placental coated vesicles contain receptor-bound transferrin.

Human placental coated vesicles have been purified by a method involving sucrose-density-gradient centrifugation and treatment with wheat-germ agglutinin. These preparations were free of contamination by placental microvillus fragments. Crossed immunoelectrophoresis demonstrated that the coated vesicles contained a single serum protein, which was identified as transferrin. This transferrin was only observed after the vesicles were treated with a non-ionic detergent, and its behaviour during crossed hydrophobic-interaction immunoelectrophoresis suggested that a large proportion of it was receptor-bound. No other serum proteins, including immunoglobulin G, could be detected in these preparations. Receptor-bound transferrin was the only antigen common to placental coated vesicles and microvilli, implying that other plasma-membrane proteins are excluded from the region of membrane involved in coated-vesicle formation.

Cell Fractionation↗

Propranolol, alprenolol and oxprenolol metabolism in the dog. Identification of N-methylated metabolites.

The metabolism of propranolol, alprenolol and oxprenolol was studied in the dog and rat; propranolol in five additional species, including man. Basic, phenolic and neutral metabolites were extracted from urine at pH 9.6 after enzymatic hydrolysis. Separation and identification of parent drug and seven metabolites each for propranolol, alprenolol and oxprenolol in the dog were accomplished by gas chromatography mass spectrometry as the trifluoroacetyl derivatives. A very uniform and predictable fragmentation pattern was observed for all 24 compounds. Seven new metabolites were identified. The metabolism of all three drugs was qualitatively the same, including N-dealkylation followed by N-methylation or deamination of the primary amines. The parent drugs as well as all of their sidechain metabolism products were also partially ring hydroxylated. N-Methylation was only found in the dog and is a minor metabolic pathway. The stereochemical composition of N-methyldesisopropylpropranolol and its immediate precursor N-desisopropylpropranolol showed a marked enrichment of the (+)-isomer.

Alprenolol↗

A modified paper-binding procedure for the assay of nucleus-associated protein phosphokinases.

Previously existing paper-binding assay procedures gave results with large variations when employed for the measurement of nucleus-associated protein phosphokinase activities. However, a modified method, utilizing the binding of 32P-labeled phosphoprotein substrates to paper and employing washing procedures in 20% trichloroacetic acid at 60 degrees to 70 degrees C, gave highly reproducible results. This modified procedure was satisfactory with either chromatin or a nonhistone protein fraction derived therefrom as a source of enzyme, and dephosphophosvitin, lysine-rich histones, or casein as phosphoprotein substrates.

Animals↗

The elevated requirement for methionine by transformed rat liver epithelial cells in vitro.

Growth constants appear to constitute a useful tool in comparing the behavior for transformed and untransformed cells in culture. The substitution of homocysteine for methionine in culture medium caused a greater depression of growth with 3/4 of the transformed epithelial and with 10/14 other cell lines tested compared to the growth depression seen in similar untransformed lines. The greater growth depression exhibited by transformed lines on methionine-deficient, homocysteine-supplemented medium could generally be attributed to: 1. Greater growth of transformed cells in complete medium, and 2. The lower levels of methyltransferase generally found in the transformed lines. Finally methyltransferase appears to be a major determinant of growth of both normal an transformed cells growing in a methionine-deficient, homocysteine-supplemented medium.

5-Methyltetrahydrofolate-Homocysteine S-Methyltran↗

Testosterone and UVL-B stimulation of epidermal melanocytes in rat scrotal skin.

The effects of UVL-B and/or testosterone replacemnt therapy are compared in normal and castrated rats in order to determine whether testosterone is required for UVL-B (290-315 nm) stimulation of melanogenesis in the testosterone-dependent epidermal melanocyte system of the scrotal skin of black Long Evans rats. Testosterone is not a prerequisite for UVL-B stimulation of melanocytes as in both castrates and normal animals the melanocytes respond to UVL-B by increases in size, length and number of dendrites (dendriticness), and tyrosinase activity (intensity of Dopa reaction). Addition of testosterone to castrates does enhance the effects of UVL-B. However, UVL-B with or without testosterone cannot maintain normal melanogenesis in rats irradiated immediately after castration nor can it restore normal melanogenesis following long term castration. Bth the amount of UVL nergy/exposure and the number of exposures are important variables in stimulation of the epidermal melanocytes. Administration of a dose of UVL-B to castrates in a single exposure is ineffective, while the same overall dose spread over several exposures increases the size and dendriticness of melanocytes. Testosterone and UVL-B act synergistically in affecting melanogenesis although neither singly nor in combination are they able to fully restore normal melanogenesis.

Animals↗

Effects of polyamines on prostatic chromatin- and non-histone-protein-associated protein kinase reactions.

Studies are presented on the influence of polyamines on prostatic chromatin- and non-histone-protein-associated protein kinase reactions involving both exogenous and endogenous substrates. The activities toward the model acidic protein substrate, dephosphophosvitin, were maximal at 160--200mM-NaCl (or -KCl or -NH4Cl). Under these conditions, spermidine and spermine added in concentrations up to 2mM were essentially without effect. However, without addition of NaCl to the medium, marked stimulation of these reactions was elicited by these polyamines at 1--2mM concentrations. The stimulatory effects were not due to non-specific changes in the ionic strength or to substitution of spermine for Mg2+, as maximal stimulation by 1 mM-spermine was observed only at optimal (2--4mM) Mg2+ concentrations. Qualitatively similar effects of polyamines were observed with enzyme preparations from the prostates of castrated rats, and with chromatin and non-histone-protein preparations from other tissues besides ventral prostate. When phosphorylation of endogenous non-histone proteins of the chromatin was measured, spermine stimulated both the initial rates and the final extent of transphosphorylation, even in the presence of optimal concentration of NaCl. By contrast, spermine or spermidine had no effect on the chromatin- and non-histone-protein-associated protein kinase reactions determined with lysine-rich histones as substrates. Chemically NN-dimethylated dephosphophosvitin was a less active substrate for the chromatin-associated protein kinase, but its phosphorylation was more markedly stimulated by spermine in comparison with unmodified dephosphophosvitin. These observations hint that the polyamine stimulations of the various protein kinase reactions may be due to effects on the conformations of the non-histone protein substrates rather than on the kinases themselves.

Animals↗

Acidic-phosphoprotein phosphatase activity of rat ventral prostate nuclei: apparent lack of effect of androgens.

A protein phosphatase activity has been demonstrated in nuclei of rat ventral prostate utilizing 32P-labelled phosvitin as a model acidic phosphoprotein substrate. This phosphoprotein phosphatase has a pH optimum of 6.7, is unaffected by the sulphydryl protecting agent 2-mercaptoethanol, and requires a divalent cation for maximal activity. Of the various divalent cations tested, Mg2+ is the most effective in reactivating the EDTA-inhibited enzyme. The phosphatase is inhibited by sodium flouride, sodium oxalate, N-ethylmaleimide, ATP and ADP but is relatively insensitive to ammonium molybdate. Increased ionic strength of the reaction medium also causes a reduction in the enzyme activity, e.g., by 48% at 200 mM sodium chloride. The activity of the acidic phosphoprotein phosphatase did not change significantly at 48 h or 96 h post-orchiectomy when expressed per unit of nuclear protein. However, it is reduced by approx. 30% at these times after castration if based on DNA content. The decline in activity per nucleus reflects the decrease in the realtive nuclear protein content observed at 48 h or 96 h post-orchiectomy. This suggests that the decline in the phosphorylation of prostatic nuclear acidic proteins which occurs upon androgen withdrawal is not due to increased nuclear phosphatase activity.

Androgens↗