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Biomedical subjects

M Moreau

Publications and source records attributed to M Moreau.

At least 127 records · Page 7Linked to original sources

A transpedicularly implanted anterior spinal support.

This article describes a device that has been developed to augment posterior spinal fracture fixation by providing support for the anterior column of the spine. The device is contracted for insertion through the limited opening provided by the transpedicular approach to the vertebral body. Once in situ it is configured to support physiological loads. In this way the structural limitations of posterior instrumentation and the surgical difficulties of anterior spinal fixation instrumentation are overcome. A preliminary investigation of the new device together with in vitro studies of a prototype are described in this report. In vitro tests to determine the expansion characteristics and structural performance of the device were performed together with tests to determine the efficacy of the support to reduce loading on the posterior fixation device. It is shown that the device has the necessary expansion characteristics and can support physiological loads without failure. Tests using porcine vertebral columns with a simulated vertebral fracture showed a significant reduction in load carried by the posterior fixation plates (44 +/- 16%) when the anterior support was included in the construct. In addition, the difference in loading between the left and right plates was significantly reduced (81 +/- 12%).

Bone Cements↗

Expression of membrane targeted aequorin in Xenopus laevis oocytes.

We described here a system for high level of expression of the calcium activated photoprotein aequorin. This protein has been targeted to the plasma membrane of Xenopus oocyte by nuclear microinjection of a plasmid containing a construction of a chimeric cDNA encoding a fusion protein composed of the photoprotein aequorin and the 5-HT1A receptor. The expression of this fusion protein is placed under the control of RSV promoter. Functional photoprotein was reconstituted in the oocyte by incubation with coelenterazine. The amount of photoprotein 24 h after nuclear microinjection of the plasmid was sufficient to trigger a detectable light emission following calcium entry. The efficiency of the expression is correlated with the dose of plasmid injected. Intracytoplasmic injection of the plasmid always failed in photoprotein expression. Targeting of the apoprotein was demonstrated by immunolocalization under confocal microscopy. In our experimental conditions, the apoprotein was always localized at the animal pole above the nucleus. We never observed expression and targeting to the plasma membrane of the vegetal pole. WE suggest that such expression might be of great interest for the study of numerous problems of developmental biology, in which calcium-dependent pathways are involved.

Aequorin↗

Expression of L-type Ca2+ channel during early embryogenesis in Xenopus laevis.

The mechanisms involved in the first step of neurogenesis, i.e. neural induction, are poorly understood, particularly in terms of the signalling pathway. In a recent work it has been shown that in urodeles the activation of L-type calcium channels is sufficient to trigger neural induction. In order to substantiate a possible role of this channel in early development in anurans, we have detailed the kinetics of the expression and the localization of the alpha 1 subunit of L-type calcium channel in the early stages of Xenopus laevis embryogenesis using immunological techniques. We observed that the expression of the alpha 1 subunit started during blastulation, where a cytoplasmic labeling was observed. At the onset of gastrulation alpha 1 was targeted to the plasma membrane of the dorsal and the ventral ectoderm. Some labeling was found in the mesoderm but never in the endoderm. This expression seems to be general, since similar results have been obtained in anurans (Xenopus) and in urodeles (Pleurodeles). In addition, we found that the alpha 0 subunit of the G(o) protein is expressed simultaneously and strictly colocalized with the alpha 1 subunit of the L-type calcium channel. The role of this channel and its regulation by G(o) protein during early neurogenesis is discussed.

Animals↗

Increased internal Ca2+ mediates neural induction in the amphibian embryo.

The molecular mechanism of neural induction is still unknown and the identity of the natural inducer remains elusive. It has been suggested that both the protein kinase C and cAMP signal transduction pathways may be involved in mediating its action. Here we provide evidence that Ca2+ is implicated in the process of transduction of the neuralizing signal. We find that an increase in intracellular Ca2+ concentration [Ca2+]i occurs during neural induction provoked in vitro by the lectin Con A in Pleurodeles waltl embryo. We demonstrate that specific L-type Ca2+ channel agonists also trigger neural induction. Conversely, noninducing lectins do not raise [Ca2+]i. Ryanodine and caffeine trigger neural induction. An increase in [Ca2+]i was also observed after treatment with the phorbol 12-myristate 13-acetate, which has been reported to be inductive. The [Ca2+]i increase triggered by phorbol ester and Con A was abolished by staurosporine and by L-type Ca2+ channel antagonists. Our findings demonstrate that the [Ca2+]i increase occurs via L-type Ca2+ channels. We suggest an amplification of this increase by a Ca(2+)-induced Ca2+ release mechanism which involves intracellular ryanodine-sensitive stores. We propose that Ca(2+)-dependent processes controlled by protein kinase C are implicated in the regulation of gene expression in response to neural induction.

Alkaloids↗

Recruitment of plasma membrane voltage-dependent calcium-permeable channels in carrot cells.

Numerous biological assays and pharmacological studies have led to the suggestion that depolarization-activated plasma membrane Ca2+ channels play prominent roles in signal perception and transduction processes during growth and development of higher plants. The recent application of patch-clamp techniques to isolated carrot protoplasts has led to direct voltage-clamp evidence for the existence of Ca2+ channels activated by physiological depolarizations in the plasma membrane of higher plant cells. However, these voltage-dependent Ca2+ channels were not stable and their activities decreased following the establishment of whole-cell recordings. We show here that large pre-depolarizing pulses positive to 0 mV induced not only the recovery of Ca2+ channel activities, but also the activation of initially quiescent voltage-dependent Ca2+ channels in the plasma membrane (recruitment). This recruitment was dependent on the intensity and duration of membrane depolarizations, i.e. the higher and longer the pre-depolarization, the greater the recruitment. Pre-depolarizing pulses to +118 mV during 30 s increased the initial calcium currents 5- to 10-fold. The recruited channels were permeable to Ba2+ and Sr2+ ions. The data suggested that voltage-dependent Ca(2+)-permeable channels are regulated by biological mechanisms which might be induced by large pre-depolarizations of the plasma membrane. In addition, this study provides evidence for the existence in the plasma membrane of higher plant cells of a large number of voltage-dependent Ca2+ channels of which a major part are inactive and quiescent. It is suggested that quiescent Ca2+ channels can be rapidly recruited for Ca(2+)-dependent signal transduction.

Calcium↗

Reception and transduction of the serotonin signal responsible for meiosis reinitiation in oocytes of the Japanese clam Ruditapes philippinarum.

Prophase-arrested oocytes of Ruditapes philippinarum are triggered to undergo germinal vesicle breakdown under the influence of the neurohormone serotonin (5HT) and then arrest in metaphase 1. Our data show that these oocytes possess a single class of original 5HT receptors. Their binding parameters have been determined on semipurified membrane preparations incubated with [3H]5HT. No significant differences were observed when comparing 5HT-competent and -incompetent batches as well as prophase- or metaphase-arrested oocytes. Specific experiments including incubation with mastoparan or mas 7, GTP iontophoresis, and IP3 quantification strongly suggest that these receptors must be coupled with G-proteins to be functional. Peak change in IP3 mass occurs at 3 min and is likely to trigger the 5HT-dependent Ca2+ transient that begins at this time. In metaphase-arrested oocytes, binding of 5HT to its receptors no longer produces a Ca2+ surger. This is likely to result from a negative retrocontrol loop which would involve kinase C and exert its effect upstream of the Ca2+ surge. Indeed, the phorbol ester PMA proved able to reduce the Ca2+ response and to block 5HT action when applied during the first 3 min corresponding to the hormone-dependent period. Such an inhibition was reversed in the presence of 5 microM of the C kinase inhibitor GF109203X and could be bypassed by ionophore, ammonia, and thapsigargin, which trigger a receptor-independent Ca2+ surge.

Animals↗

Presence of leukaemia inhibitory factor and interleukin 6 in porcine uterine secretions prior to conceptus attachment.

Leukaemia inhibitory factor (LIF) plays an important role in embryo development and implantation. We detected peak LIF activity in porcine uterine luminal fluids (ULF) at day 12 of gestation and during day 7 and 13 of the oestrous cycle. A radio-receptor competition assay showed the presence of a molecule in ULF specifically binding to human LIF receptor (LIF-R). LIF activity was partially neutralized by anti-human LIF antibody. Interleukin-6 (IL-6) activity was detected in ULF throughout the oestrous cycle and pre-implantation period. An anti-murine alpha chain (gp80) of IL-6 receptor (IL-6R) specifically neutralized this activity. LIF and IL-6 mRNA were only detected in day 11 endometrium. The presence of LIF or IL-6 in the uterine cavity has not been previously reported. Our results extend LIF production by endometrium during the oestrous cycle and pre-implantation period to another mammalian species other than mouse.

Animals↗

Troubleshooting implantable cardioverter defibrillator system malfunctions: the role of impedance measurements.

High impedance measurements may be used to troubleshoot ICD system malfunction. In four different cases a defective system was identified or confirmed by an abnormal impedance: two secondary to lead fractures, one to an adapter faulty connection, and one to a loose set-screw connection. This led to further diagnostic procedures that ultimately localized the difficulty. Though the problems with ICD may be of diverse origins, routine use of impedance measurement for the troubleshooting of these systems may serve as an early sign of malfunction and would point the need for further investigation.

Aged↗

Dual-energy x-ray imaging technique for in vitro tissue composition measurement.

A dual-energy in vitro radiographic technique has been developed to study the thickness of tissue and bone within atherosclerotic plaques. Results concerning the accuracy and precision of the thickness measurements using this technique are presented and discussed. Planar radiographs of phantoms were obtained with a low-energy spectrum (45 kVp, no added filtration) and a high-energy spectrum (100 kVp, 2.88-mm copper-added filtration), and then decomposed into bone-equivalent and Lucite basis-material images. Thickness measurements from these images yielded average accuracies of +/- 750 microns for the Lucite images, and +/- 25 microns for the bone-equivalent images. The imprecision (one standard deviation) of the thickness measurements was +/- 192 and +/- 47 microns for the Lucite and the bone-equivalent images, respectively (for thin sections). Although the accuracy and precision of Lucite thickness measurements were not as good as those obtained with other techniques, such as the iodine displacement technique, the accuracy and precision of the bone thickness measurements are shown to be much better. The high accuracy and precision of the bone measurement makes dual energy a very appealing technique for analyzing the physical properties of calcified atherosclerotic plaques in excised arterial specimens.

Absorptiometry, Photon↗

[Update on menopause].

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Estrogen Replacement Therapy↗

Evidence for the involvement of internal calcium stores during serotonin-induced meiosis reinitation in oocytes of the bivalve mollusc Ruditapes philippinarum.

In contrast to the situation found in the bivalves Barnea candida and Spisula solidissima, prophase-arrested oocytes of Ruditapes philippinarum cannot be fertilized when removed from the ovary. They must first undergo germinal vesicle breakdown under the influence of the neurohormone serotonin (5-HT), which drives them to a second block occurring in metaphase of the first maturation division. In the studies described in this paper, we investigate the possibility that calcium is involved as a second messenger in controlling this first step in the reinitiation of meiosis. Our data show that, in addition to 5-HT, ionophore, thapsigargin, and the weak bases ammonia and procaine can also induce prophase-arrested oocytes of Ruditapes to resume meiosis. 5-HT, thapsigargin, and ammonia all trigger a surge of intracellular Ca2+ and are effective even in the absence of external Ca2+. That such Ca2+ transients, which are enhanced in the presence of external Ca2+, actually play a key role in the process of meiosis reinitation is shown by the fact that loading the oocytes with BAPTA/AM or treating them with D-600 blocks maturation. In contrast, excess KCl, which has been shown to trigger meiosis reinitiation of prophase-arrested oocytes of Barnea and Spisula and to activate metaphase I-arrested oocytes of Ruditapes, does not produce any significant intracellular Ca2+ transient nor does it reinitiate meiosis, when added to Ruditapes prophase-arrested oocytes. These data suggest that such voltage-operated Ca2+ channels may only appear during the course of maturation and that both intracellular and extracellular Ca2+ are involved in triggering 5-HT-dependent release from the prophase block in this species.

Animals↗

Effects of PDE inhibitors and carbachol on the L-type Ca current in guinea pig ventricular myocytes.

The phosphodiesterase inhibitors 3-isobutyl-1-methylxanthine (IBMX; 100 microM) and papaverine (100 microM) increased peak L-type Ca current (ICa) more than fivefold in a way similar to isoproterenol, forskolin, or intracellular adenosine 3',5'-cyclic monophosphate in guinea pig ventricular myocytes studied with the whole cell voltage-clamp technique at 22-24 degrees C. IBMX and papaverine could also induce a chloride current. Both drugs caused an apparent increase of ICa inactivation as revealed by 1) a negative shift of the ICa inactivation curve between -40 and 0 mV and 2) a suppression of the relief from inactivation at potentials positive to 0 mV. In the presence of IBMX or papaverine, the amplitudes of both the rapidly and slowly inactivating components of ICa were increased; the effect on the fast component was more pronounced. The drugs did not accelerate the inactivation time course of either component. Carbachol (CCh; 100 microM) reversed the increase in ICa produced by IBMX or papaverine. However, ICa could not be restored to its original magnitude on washout of CCh in the presence of phosphodiesterase inhibitors. In pertussis toxin-treated cells or in the presence of Ly-83583 (1-100 microM), IBMX retained its effect but CCh was unable to reduce ICa. Dialysis with guanosine 3',5'-cyclic monophosphate (cGMP; 0.1-100 microM) or 8-bromoguanosine 3',5'-cyclic monophosphate (30 microM) suppressed the increase of ICa by IBMX; the inhibition by cGMP was additive with that produced by CCh. We suggest that the major part of IBMX and papaverine effect is mediated by phosphodiesterase inhibition and involves an increase in intracellular adenosine 3',5'-cyclic monophosphate levels. CCh reversal of phosphodiesterase inhibitor action probably involves an elevation of cGMP levels and activation of cGMP-dependent protein kinase.

1-Methyl-3-isobutylxanthine↗