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Biomedical subjects

A Defontaine

Publications and source records attributed to A Defontaine.

7 recordsLinked to original sources

Oligonucleotide quartz crystal microbalance sensor for the microalgae Alexandrium minutum (Dinophyceae).

We report the immobilization on a gold surface of a 20-base DNA probe labeled with disulfide group and on the selective hybridization with the complementary 20-base DNA strand. The oligonucleotide probe is the complementary strand of a partial sequence of the gene encoding for a large ribosomal RNA sub-unit which is a coding sequence of Alexandrium minutum DNA, a microalgae that produces neurotoxins responsible for paralytic shellfish poisoning on European and Asian coasts. The kinetics of DNA probe immobilization and hybridization were monitored in situ by using a 27 MHz quartz crystal microbalance under controlled hydrodynamic conditions. The frequency of the setup is stable to within a few hertz, corresponding to the nanogram scale, for 3h and makes it possible to follow frequency change from immobilization of the probe to hybridization of the complementary DNA target. This setup constitutes a biosensor, which is sensitive and selective, and the hybridization ratio between hybridized complementary DNA and immobilized DNA probes is 47%.

Animals↗

Multi-formalism modelling and simulation: application to cardiac modelling.

Cardiovascular modelling has been a major research subject for the last decade. Different cardiac models have been developed at a cellular level as well as at the whole organ level. Most of these models are defined by a comprehensive cellular modelling using continuous formalisms or by a tissue-level modelling often based on discrete formalisms. Nevertheless, both views still suffer from difficulties that reduce their clinical applications: the first approach requires heavy computational resources while the second one is not able to reproduce certain pathologies. This paper presents an original methodology trying to gather advantages from both approaches, by means of a hybrid model mixing discrete and continuous formalisms. This method has been applied to define a hybrid model of cardiac action potential propagation on a 2D grid of endocardial cells, combining cellular automata and a set of cells defined by the Beeler-Reuter model. For simulations under physiological and ischemic conditions, results show that the action potential propagation as well as electrogram reconstructions are consistent with clinical diagnosis. Finally, the advantage of the proposed approach is discussed within the frame of cardiac modelling and simulation.

Action Potentials↗

Bacterial production and purification of recombinant human prolactin.

Escherichia coli cells transformed with a recombinant plasmid (pT7L) containing the coding sequence of human prolactin (hPrl) expressed a new protein. This protein, comigrating with human Prl on sodium dodecyl sulfate (SDS)-polyacrylamide gels, represented 50% of the total bacterial extract. Immunoprecipitation of [35S]methionine-labeled bacterial lysate with a rabbit antiserum to hPrl followed by SDS-polyacrylamide gel electrophoresis (PAGE) analysis showed that the major component had a Mr identical to that of standard hPrl. The majority of the recombinant hPrl (r-hPrl) accumulated in inclusion bodies. Analysis of these inclusion bodies by SDS-PAGE under nonreducing conditions showed that they are composed mostly of fully reduced monomers. Solubilization of the inclusion bodies and protein denaturation were performed in 8 M urea. Refolding during the renaturation procedure was confirmed by SDS-PAGE under nonreducing conditions. r-hPrl was further purified by gel permeation chromatography on a fast protein liquid chromatography column. More than 95% of the molecules were recovered as oxidized monomeric forms. The refolded molecule was tested for its bioactivity in the Nb2 lymphoma mitogenic assay. The dose-response curves obtained with either r-hPrl or pituitary-derived hPrl showed a complete parallelism. Furthermore, Nb2 cell proliferation was completely blocked by addition of hPrl antiserum to both preparations. Recombinant hPrl is identical to natural hPrl except for an additional methionine group at the amino terminal end.

Amino Acid Sequence↗

Transient hypoparathyroidism induced by synthetic human parathyroid hormone-(1-34) treatment.

Daily injections of low doses of a synthetic fragment of human PTH [hPTH-(1-34) have increased iliac trabecular bone volume when used in the treatment of osteoporosis. In approximately 50 patients no major side-effects had occurred. However, during daily sc 100-micrograms injections of the peptide, one patient repeatedly developed parathyroid hypofunction which resolved each time treatment was stopped. Specific immunoglobulin G (IgG) antibodies binding [125I]hPTH-(1-34) were identified in the patient's serum, and positive immunohistochemical reactions were obtained when bovine parathyroid sections were exposed to the patient's IgG. After adsorption with PTH, the patient's IgG, free of anti-PTH antibodies, reacted with renal cell membranes, as demonstrated by indirect immunofluorescence and blocked renal PTH-dependent adenylate-cyclase activation in vitro. These results support the hypothesis that anti-PTH receptor as well as anti-PTH antibodies were generated during hPTH-(1-34) treatment, which led to the development of hypoparathyroidism when their titers were high.

Adenylyl Cyclases↗

Luteinizing hormone and prolactin release by cultured pituitary cells and by gonadotrope- and lactotrope-enriched populations in the presence of modulators of adenylate cyclase.

The effects of 3 compounds previously known to alter the adenylate cyclase activity in various tissues were tested on rat pituitary. Forskolin increased while RMI 12,330 A and SQ 22,536 decreased enzyme activity of pituitary homogenates. Forskolin (10(-5) M) promoted in cultured cells a massive synthesis of cyclic adenosine monophosphate (cAMP) and intense output of the nucleotide in the medium; at the same concentration, RMI 12,330 A decreased the cell content of cAMP while release was unaffected, and SQ 22,536 exerted no effect on the cyclic nucleotide amount of both cell and medium. The basal release of plated total cells or enriched gonadotropes was slightly stimulated by forskolin (10(-7) - 10(-5) M) though the level of stimulation never attained the one produced by 10(-7) M luteinizing hormone-releasing hormone (LHRH); it was affected neither by RMI 12,330 A (10(-7) - 10(-5) M) nor by SQ 22,536 (10(-7) - 10(-4) M). The LHRH-stimulating effect was slightly amplified during the 1st h (sensitivity response of cells) by low concentrations of forskolin (10(-7) and 10(-6) M) but the overall luteinizing hormone (LH) release after 4 h of incubation (capacity response of cells) was unmodified. Higher concentrations of the diterpene inhibited the LHRH effect as seen with total cells and with enriched gonadotropes. RMI 12,330 A at 10(-5) M significantly inhibited the LHRH-promoted LH release from total cells and from gonadotropes but a higher concentration (10(-4) M) promoted a large unspecific release that masked the LHRH effect. SQ 22,536 never modified the cell response to LHRH, whatever the drug concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenine↗

[The Orf nodule].

Human Orf disease is an exceptional dermatologic benign condition, due to a Parapox virus. In sheep and goats this infection is termed "Ecthyma Contagiosum". Human beings are contaminated from infected animals. We report three cases of Orf disease in the same family. Typical viral particles have been identified in the skin of one of these patients by electronic microscopy but cultures failed to recover the pathogen. A complete study of the literature allows us to review current knowledge on this disease with which practitioners are unfamiliar.

Adolescent↗