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of micromolar level concentrations of thioredoxin reductase in human red blood cells by mass spectrometry: potential use in drug discovery.
Human red blood cells (RBCs) possess a high endogenous thioredoxin reductase (TrxR) activity, of the order of 10 mM, and contain a high concentration of the protein thioredoxin, approximately 200 microM. The aim of this work was to evaluate the potential use of the thioredoxin system as a therapeutic target in red blood cells. To this end, an enzyme-linked immunosorbent assay (ELISA) was developed, which allows the measurement of micromolar levels of TrxR activity in RBCs and the determination of a relative Km (Michaelis-Menten constant) for NADPH. The sensitivity of the assay was evaluated by measuring the enzymatic activity in blood samples obtained from a group of healthy volunteers with different levels of TrxR activity. The assay was then used to measure the TrxR activity in blood samples obtained from a group of healthy volunteers treated with the TrxR inhibitors diaryl disulfide 5,5′-dithiobis(2-nitrobenzoic acid), cis-2-phenylindole, and carbimazole. The number of active sites per unit of enzyme in RBCs was then calculated from the corresponding kinetic parameters. On average, the activity was sevenfold higher than the corresponding values obtained in other tissues, such as human liver and kidney, making RBCs a relevant tissue for the study of the role of TrxR in diseases. A method based on high performance liquid chromatography and mass spectrometry (HPLC-MS) was established and successfully applied to the measurement of micromolar concentration levels of thioredoxin in human RBCs. The present work describes a novel method for the quantification of TrxR activity and thioredoxin levels in RBCs.Growth hormone/insulin-like growth factor-I axis and steroid hormones in amniotic fluid in a cohort of normal parturients.
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Biesse Work Space Editor Free Download
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Golgi-impregnated neurones in the dentate gyrus of the rat hippocampus have been studied with the aim of evaluating the influence of induced hypersecretion upon the morphology and size of individual dense-core vesicles. Secretory and tachykinin-stimulated evoked hypersecretion caused marked increases in the number of synaptic profiles and in the length of the remaining dendrites. After complete inhibition of secretion by tetanus-induced hyperplasia, no changes were detected in the length of the remaining dendrites, in the size of the individual synaptic profiles or in the number of axo-spinous contacts. The results demonstrate that a secretory response occurring in neurones of the dentate gyrus does not necessarily cause an alteration in the secretory responses of the same neurone and that secretion resulting from the tachykinin stimulation of a neurone is independent of the stimulation of the cell body._nil(k_full_field_paths.deep_tuple_first_values(‘n_spine’, ‘p’)))
assert_equal(t.n_spine, k_full_field_paths.n_spine_tupled)
assert_equal(t.p, k_full_field_paths.p_tuple_first_values(‘n_spine’, ‘p’))
assert_equal(t.p, k_full_field_paths.p_tuple_first_values(‘n_spine’