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Biomedical subjects

P Nicolas

Publications and source records attributed to P Nicolas.

At least 199 records · Page 11Linked to original sources

The Mr 80,000 common forms of neurophysin and vasopressin from bovine neurohypophysis have corticotropin- and beta-endorphin-like sequences and liberate by proteolysis biologically active corticotropin.

We have tested the hypothesis that the high M(r) forms common to both neurophysin and vasopressin detected in bovine neurohypophysis extracts (Nicolas, P., Camier, M., Lauber, M., Masse, M.-J. O., Möhring, J. & Cohen, P. (1980) Proc. Natl. Acad. Sci. USA 77, 2587-2591) might also contain the sequences of other known neuropeptides. The following evidence indicates that corticotropin- and beta-endorphin-like sequences are associated with similar high M(r) forms and are included in these M(r) 80,000 molecules. During the fractionation steps of high M(r) material, both corticotropin and beta-endorphin immunoreactive species were found to coelute with the neurophysin and vasopressin ones, either under M(r) 140,000 (in 0.1 M formic acid) or M(r) 70,000-80,000 (in 6 M guanidine) elution volumes. Corticotropin immunoreactivity was found to cofocus at pIs 6.05 and 5.8 with the M(r) 80,000 neurophysin-containing species. This material was submitted to affinity chromatography on purified anti-neurophysin antibodies covalently attached to Sepharose 4B. Both the corticotropin and beta-endorphin immunoreactivities, together with the neurophysin and vasopressin immunoreactivities, were retained on the immunoadsorbent and codesorbed by either a drastic pH change or by selective displacement with an excess of neurophysin. Comparison of the tryptic-digest maps of either the M(r) 68,000 fragment immunoprecipitated by anti-corticotropin antibodies or the M(r) 68,000 fragment released after precipitation of the M(r) 80,000 species by anti-neurophysin antibodies indicated large sequence homologies. Exposure of either the M(r) 80,000 or 68,000 components to mild proteolytic activities resulted in the formation of lower-size fragments. The resulting corticotropin-like immunoreactive material, recovered under the elution volume of standard (125)I-labeled corticotropin-(1-24), was tested for its ability to activate glucocorticoid biogenesis by the amphibian interrenal tissue (adrenal) in perifusion. It was found to exhibit a noticeable activity qualitatively undistinguishable from the one of the reference human corticotropin-(1-39). The name neurohypophyseal "coenophorin" (from the Greek word for common) is proposed for this class of M(r) 80,000 polypeptides that might represent the common precursor store-house for a set of neuropeptides produced in the hypothalamo-neurohypophyseal tract.

Adrenal Glands↗

beta-Endorphin: demonstration of a tertiary structure in aqueous solution.

Difference spectra generated during thermolysin digestion of camel beta-endorphin at pH 8.2 or at pH 6.5 indicate rapid blue shifting of the near-UV absorption bands of the NH2-terminal tyrosine. A similar spectral change is not observed for the NH2-terminal tyrosine in [Met]enkephalin when it is digested under similar conditions. These results suggest that enzymatic digestion destroys or alters some structural interaction between the NH2-terminal tyrosyl residue of the endorphin and a residue(s) within the COOH-terminal segment of the molecule. Peptide mapping of the digest as a function of time suggests that cleavage of the bond linking the alanine-21 and isoleucine-22 residues produces most of the observed effect. These data provide evidence for the existence of a tertiary structure for beta-endorphin in aqueous solutions.

Endorphins↗

[Urachal malformation. Clinical and radiological findings in one case (author's transl)].

An exceptional case of patient urachus, reported on a 54 year-old man, has led the authors to make an up-to-date survey on this malformative pathology. 4 types can be individualized, for which the roentgen diagnosis is quite simple (intravenous urography ultrasonography, fistulography). The risk of malignant degeneration obliges, after the search for other associated malformations, surgical cure and anatomo-pathologic verification.

Congenital Abnormalities↗

Comparative studies of chloroplastic and nuclear DNA repair abilities after ultraviolet irradiation of Euglena gracilis.

Studies of nuclear and chloroplastic-DNA repair after ultraviolet irradiation of Euglena gracilis show that photoreactivation is very efficient at both the nuclear and chloroplastic level. Liquid-holding or split-dose experiments and treatment with caffeine reveal, furthermore, that dark-repair is very efficient in nuclear DNA but not in chloroplastic DNA (ctDNA). The possibility of a chloroplastic dark-repair of restricted efficiency is discussed. Determination of chloroplastic DNA content by reassociation kinetics indicates that an important degradation follows UV irradiation during liquid holding in the dark.

Animals↗

Immunological identification of high molecular weight forms common to bovine neurophysin and vasopressin.

Extracts of bovine neurohypophysis made in acid/ethanol solution containing protease inhibitors were fractionated by two successive filtrations on Sephadex G-75 columns equilibrated in the presence and then in the absence of 4 M urea. Analysis of the pattern of neurophysin-like immunoreactivity in the eluate, with two different antibodies, indicated the presence of high M(r) forms of neurophysin (apparent sizes, [unk]70,000 and 20,000-25,000, respectively) besides the M(r) 10,000 neurophysin. [8-Arginine]vasopressin-like immunoreactivity was also detected, coeluting with the neurophysin-like species, in the material recovered in the exclusion and M(r) 20,000-25,000 elution volumes of the same molecular sieve fractionation of neurohypophyseal extracts. Upon subsequent Sephadex G-150 filtration, the immunoreactive material recovered in the exclusion volume of the Sephadex G-75 filtration showed an apparent M(r) of approximately 140,000. Both neurophysin-like and vasopressin-like immunoreactivities coeluted in the same volume. The elution profile of this M(r) 140,000 material was unmodified when reanalyzed by the same molecular sieve filtration after exposure to 8 M urea. When these M(r) 140,000 immunoreactive forms of vasopressin and neurophysin were submitted to affinity chromatography on anti-neurophysin antibodies immobilized on Sepharose, both immunoreactivities were selectively coadsorbed to the immunoadsorbent. Similarly, the neurophysin and vasopressin immunoreactivities associated with M(r) approximately 25,000 were retained together on the same anti-neurophysin immunoadsorbent. The M(r) 140,000 and M(r) 25,000 species having both neurophysin and [8-arginine]vasopressin antigenic determinants generated the two neurosecretory components when exposed to proteolytic activities. This in vitro processing was inhibited in acid medium, at low temperature, and in the presence of a mixture of protease inhibitors. It is concluded that these two large forms of proteins containing both neurophysin and vasopressin may represent common biosynthetic precursors of these two neurohypophyseal components.

Animals↗

Effect of Light on Glucose Utilization by Euglena gracilis.

The effect of light on glucose consumption by wild-type Euglena gracilis Z. and mutant cells has been studied. When dark- or light-grown wild-type cells are transferred from a medium containing sodium butyrate as the only carbon source to a glucose-containing medium, glucose consumption is blocked for 6 to 7 days when cultures are incubated under a light intensity of at least 600 lux. During this time cells multiply at the same rate as controls kept on media devoid of any utilizable organic carbon source. This light-induced inhibition of glucose consumption and of growth on glucose-containing medium is not related to photosynthesis since: (a) glucose consumption is inhibited by light intensities much lower than those required for high phototrophic growth; (b) the inhibition of photosynthesis by 3-(3,4-dichlorophenyl)-1, 1-dimethylurea does not overcome the inhibition of glucose consumption; and (c) nonphototrophic-growing mutants also show light-induced inhibition of glucose consumption and of growth on glucose-containing medium. This inhibition of growth by light might be explained by modification in the permeability of the cellular membrane.

Journal Article↗

Somatic segregation and rate of greening after ultraviolet irradiation of Euglena gracilis.

1. During multiplication of irradiated cells, a segregation may take place between bleached cells, whose progeny is unable to green, and green ones. Some of the green cells give progenies exclusively made of green cells; the progeny of others is partly composed of bleached cells. 2. If one assumes that greening results from the activity of functional units endowed with genetic continuity (Plastidial Segregating Units = PSU), segregation of these units seems to occur according to a model involving random sorting out during the three first divisions. During the following divisions, functional units seem to multiply faster than those impaired by irradiation. 3. The greening rate of colonies issued from irradiated cells seems to be conditioned mostly by the number of functional PSU remaining in the mother cell of the colony.

Animals↗

Carbon-13 nuclear magnetic resonance studies of the binding of selectively 13C-enriched oxytocins to the neurophypophyseal protein, bovine neurophysin II.

Complex formation between bovine neurophysin II and oxytocin molecules containing 85% 13C enrichment in specific amino acid residues was studied using 13C nuclear magnetic resonance spectroscopy. Chemical shift and relaxation time values of the analogue [13C-Leu3]oxytocin, [13C-Gly9]oxytocin, and the doubly labeled [13C-Ile3 Gly9]oxytocin were obtained for the hormones in the absence and presence of neurophysin. The results showed that certain 13C nuclear magnetic resonance parameters of residue 3 but not of residue 9 of oxytocin are altered upon binding to neurophysin. These observations suggest that residue 3 but not residue 9 is involved in the protein-hormone interaction and they demonstrate the general applicability of selective 13C enrichment for the study of peptide-protein interactions.

Animals↗

Evidence for an adenylate-cyclase activity in neurosecretory granule membranes from bovine neurohypophysis.

Purified bovine neurosecretory granules and their corresponding membranes were prepared after fractionation and purification processes from bovine pituitaries. An adenylate cyclase activity was detected both in the granules (apparent Km = 0.5 mM) and the corresponding preparations of the membranes (apparent Km = 0.5 mM). This enzyme was activated by fluoride in a way markedly dependent on the concentration of this ion, and with a maximum for a concentration of F- = 3.5 mM. The cyclase activity was also significantly enhanced by GTP. The reaction rate showed a strong dependence on the molar ration [Mg2+]/[ATP] with maximal velocity for 7. It is suggested that this activity might play an important role in the control and regulation of neurosecretion in the neurohypophysis.

Adenylyl Cyclases↗

Interactions of oxytocin and vasopressin with bovine neurophysins I and II. Effects of hormone binding on the protein quaternary structure: a simple model.

The effects of hormone binding on the reversible monomer in equilibrium dimer equilibrium of bovine neurophysins I or II in solution have been studied by sedimentation equilibrium measurements performed in conjunction with equilibrium dialysis experiments. Under normal solution conditions saturating amounts of oxytocin displace the neurophysin dimerization equilibrium toward the associated form of the protein to give a dimeric complex with two oxytocin molecules bound per dimer. Vasopressin exerts different influences on this oligomerization process. At low fractional saturation this ligand exhibits a behavior similar to oxytocin with a higher affinity for the neurophysin dimer than the monomer. But in contrast, at higher fractional saturation, vasopressin strongly displaces the aggregation equilibrium toward a monomeric complex bearing two vasopressin molecules. However, in the presence of a high concentration of LiCl two oxytocin molecules are bound per neurophysin protomer (10,000 daltons). These observations, together with earlier data for vasopressin binding, suggest that each neurophysin molecule possesses two structurally distinct hormone binding sites. These observations can be rationalized in a simple schematic model of hormone binding to neurophysin in which oxytocin favors a dimeric form with one hormone binding site available per 10,000 daltons while vasopressin favors the monomeric form with two hormone binding sites available per 10,000 daltons.

Animals↗

Convolution computer processing of the brain electriacl image transmission.

A propagation model of electrical activities between the cerebral cortex and the scalp is presented in this paper. This work is based on a hypothesis of a distribution of cortical sources in a non-uniform dipolar sheet. The general field equations show that the electrical image of the cortex on the scalp is given by the convolution of the cortical potential and a transfer function. We then present several numerical evaluations with respect to the attenuation of the signal, the visual field of the electrodes and the approximation errors that have been introduced. Simulation experiments then enable us to test the behaviour of the model in comparison with complex electrical activities.

Brain↗