Biomedical subjects

A Jean

Publications and source records attributed to A Jean.

At least 37 records · Page 2Linked to original sources

PubMed1996

Identification of CYP2B14P and CYP2B16P, two apparent pseudogenes in the rat cytochrome P450 2B (CYP2B) subfamily.

Cytochrome P450 2B3 (CYP2B3) is a member of the CYP2B subfamily and is present constitutively in rat liver. During cloning of the CYP2B3 gene, several variant genomic inserts were isolated, exoncontaining fragments of which were identified by hybridizing to CYP2B3 cDNA or exonic probes. Nucleotide sequence analysis of one set of variant inserts showed them to carry exon 1 and the 5'-flanking region of a gene designated CYP2B14P. The last codon of CYP2B14P exon 1 is a TAG translation stop codon, and thus CYP2B14P is a pseudogene. Three other inserts covering 34 kb of genomic sequence together carried 9 exons and the 5'-and 3'-flanking regions of the gene designated CYP2B16P. The 5'-splice site of CYP2B16P intron 1 is inactivated by the replacement of G by T in the normally invariant GT dinucleotide. Thus, CYP2B16P also has the characteristics of a pseudogene. The identification of CYP2B14P and CYP2B16P brings to at least seven the number of genes or pseudogenes shown by nucleotide sequence analysis to belong to the rat CYP2B subfamily.

Animals↗

PubMed1996

In vitro models to study wound healing fibroblasts.

Phenotypic and contractile properties of human fibroblasts from dermis and from an experimental wound model were studied in vitro. When cultured in monolayer, dermal fibroblasts had an elongated spindle shape, were small in diameter and grew at a high rate. Wound fibroblasts grew slowly and were large, star shaped and had cytoplasmic stress fibres. Smooth muscle alpha actin was detected in 10 percent of dermal cells, whereas 20-80 per cent of wound fibroblasts contained this protein in their cytoplasm. The contractile property of cells was evaluated using a three-dimensional cell culture model. Our results show that wound fibroblasts contract collagen gels during the first days more strongly than dermal fibroblasts. These results show that, in vitro, wound fibroblasts have greater contractile capacity than dermal cells. The significant proportion of wound fibroblasts containing alpha-smooth muscle actin suggests that alpha-smooth muscle actin ratio may be related to wound contraction.

Cells, Cultured↗

PubMed1996

Developmental study of N-methyl-D-aspartate-induced firing activity and whole-cell currents in nucleus tractus solitarii neurons.

Whole-cell recordings of rat nucleus tractus solitarii (NTS) neurons were performed on a slice preparation. We investigated possible postnatal changes in firing activities and currents induced by N-methyl-D-aspartate (NMDA) application. A total of 42 neurons were selected and fell into the following age groups: 0-5 days (n = 15), 10-15 days (n = 9) and 30-60 days (adult, n = 18). During this period, input resistance and spike duration decreased by approximately 40%. At all ages, bath application of NMDA elicited a bursting firing activity when the membrane potential was held between -60 and -75 mV. However, in the youngest cells the rhythmic bursting activity was irregular and was characterized by a progressive firing inactivation during a burst. In a tetrodotoxin-containing saline, NMDA-induced oscillations of membrane potential were retained in all age groups. The membrane current-voltage relationship of the NMDA-induced inward current (INMDA) was characterized by a region of negative slope conductance which was similar in all age groups. Thus the voltage-dependent block of INMDA is present in NTS neurons from birth, allowing NTS neurons to display membrane potential oscillations. However, postnatal maturation of repolarizing conductances, as suggested by changes in spike characteristics, could render the oscillatory activity more stable than at birth.

Aging↗

PubMed1996

Digital image ratio: a new radiographic method for quantifying changes in alveolar bone. Part 1: Theory and methodology.

A new world, digital image ratio (DIR), has been developed for directly measuring changes in alveolar bone. The image on the computer monitor represents the relative mass change between two radiographs. Fourier filtering is used to reduce noise artefacts. This method is validated through an experiment with a step wedge. DIR needs only a preliminary calibration of the experimental conditions of operation and avoids tedious calibrations for each measurement as in the case of digital image substraction. Low-voltage X-ray techniques are suggested for long-term quantitative studies of patients to minimize irradiation doses.

Absorptiometry, Photon↗

PubMed1996

Digital image ratio: a new radiographic method for quantifying changes in alveolar bone. Part II: Clinical application.

As reported in a previous paper (1) we have developed a new technique, Digital Image Ratio (DIR), which theoretically avoids some of the drawbacks of quantitative digital substraction radiography. DIR allows the direct computation and visualization of bone-mass-ratio changes. This second paper describes the use of DIR analysis to examine 20 sites in 8 patients undergoing regenerative periodontal therapy. Standardized reproducible radiographs of these 20 sites were taken before and 12 months after surgery. Ten experimental sites were treated with bone graft substitutes (natural coral or natural coral+collagen), and 10 control sites by debridement alone. None of the experimental sites had a density ratio below 1, where 1 indicates no change. The error was +/- 0.07 (0.93-1.07). The experimental sites showed an 18% mean increase in bone density (1.18), which increased to 23% (1.23) for sites filled with natural coral alone. All the control sites had values close to 1.00 (1.00 +/- 0.07) except for 3 sites, which showed a 9-15% loss of bone density. It is thus possible to compare and quantify the changes in experimental and control sites in the same patient using the percentage gain or loss of bone density. This demonstrates that DIR is suitable for clinical applications, and can be used in clinical analysis when bone changes are expected.

Alveolar Process↗

PubMed1995

Synaptic organization of the interstitial subdivision of the nucleus tractus solitarii and of its laryngeal afferents in the rat.

The nucleus tractus solitarii, the first central relay for gustatory and a variety of visceral afferents, is also an integrative center for numerous functions. Its interstitial subdivision is involved in swallowing and respiratory reflexes. The ultrastructural characteristics of this subdivision and of its laryngeal afferents were investigated in adult rat by a serial-section study and by application of wheat germ agglutinin-horseradish peroxidase conjugate to the peripheral afferent fibers. The interstitial subnucleus contained scattered small neuronal cell bodies with such ultrastructural features as a large nucleus with deep indentations and an organelle-poor cytoplasm. On the basis of their size and vesicular content, the axon terminals were classified into three categories. Group I and group II terminals were small or large, respectively, and contained mainly small, round, and clear synaptic vesicles. Group III terminals were also small but contained small, pleomorphic, and clear vesicles. Axodendritic synapses were the most numerous. They were either asymmetrical, comprised of group I and II terminals, or symmetrical, comprised of group III terminals. More than 50% were part of complex synaptic arrangements in the form of rosettes or glomeruli. Axosomatic contacts involved both group I and group III terminals and were always symmetrical. A high frequency of axoaxonic synapses was found. They were symmetrical, comprised of group III terminals on group I or II terminals. Different types of symmetrical synaptic contacts made by dendrites were also found. This study indicates also that the ipsilateral interstitial subdivision constitutes the preferential site of termination for superior laryngeal afferents. The labeled axon terminals belonged exclusively to groups I and II and were involved in both axodendritic and axoaxonic synapses. Some of the axodendritic synapses were part of rosettes or glomeruli. All these synaptic arrangements may be considered a morphological substrate for important processing of afferent information in the nucleus tractus solitarii. They may account for some of the integrative functions of the interstitial subnucleus such as physiological processes triggered from the superior laryngeal nerve.

Afferent Pathways↗

PubMed1995

Postnatal changes in glutamate binding in the lower medulla of the rat.

Changes in [3H]glutamate (Glu) binding occurring in the lower medulla of the rat between birth (P0) and adulthood (P72) were investigated on cryostat sections using in vitro receptor autoradiography. Densitometric measurements were performed in both autonomic (nucleus tractus solitarii, dorsal motor nucleus of the vagus nerve, ventrolateral medulla) and non-autonomic (inferior olive, spinal trigeminal nucleus) medullary regions. In all these areas, binding densities peaked at P9 and then gradually declined. Values close to those measured in adult animals were reached by P23-P30. These data indicate that glutamatergic neurotransmission within the lower medulla undergoes developmental changes during the early postnatal period. They thereby suggest that the neural circuits essential in organizing autonomic functions are still immature at birth.

Age Factors↗

PubMed1994

Human wound healing fibroblasts have greater contractile properties than dermal fibroblasts.

Contractile and phenotypic properties of human fibroblasts from healing wounds were compared to those of dermal fibroblasts using in vitro models. Wound fibroblasts were recovered from implants, made of a polyvinyl alcohol sponge threaded into a perforated silicone tube, 12 days after their subcutaneous implantation in human volunteers. Dermal fibroblasts were isolated from the skin of healthy subjects. Two morphologically different fibroblast populations were observed in cells cultured from implants. In order to characterize these fibroblast populations, intracellular alpha-actin expression was studied by immunofluorescence labeling of cells cultured in monolayer. This protein was detected in less than 1% of the dermal fibroblasts. By contrast, 30 to 40% of wound fibroblasts were labeled and contained fiber networks of alpha-actin. These results confirm the presence of myofibroblasts in human wound healing tissues. The contractile property of fibroblasts and myofibroblasts was evaluated using a three-dimensional cell culture model (fibroblast populated collagen gels). Cells were incorporated in a collagen matrix and cultured for 14 days. The surface area of collagen gels was measured every day. Our results show that wound fibroblasts strongly contract collagen gels during the first 24 hr (surface area at 24 hr = 20-55% of initial surface area) in comparison to dermal fibroblasts (surface area at 24 hr = 70-75% of initial surface area). This superior level of contraction was observed until the fifth day of culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗

PubMed1994

Comparative effect of calcium hydroxide and hydroxyapatite on the cellular activity of human pulp fibroblasts in vitro.

When, in vivo, calcium hydroxide [Ca(OH)2] or hydroxyapatite are used as dental pulp-capping agents, a reparative dentine bridge is observed. New hard tissue is formed directly on the hydroxyapatite, whereas a characteristic necrotic area appears under Ca(OH)2. The differing pulpal reactions to these two capping agents suggest differing cell responses. After isolation and selection of human pulp fibroblasts in vitro, the cells were characterized by their morphology, their high alkaline phosphatase specific activity, and their synthesis of type I and III collagens and fibronectin. They were then incubated in the presence of either hydroxyapatite (1 mg/ml) or Ca(OH)2 (0.8 mg/ml). With Ca(OH)2, the cells exhibited dramatical alterations in morphology, DNA synthesis, alkaline phosphatase activity and protein synthesis, in accordance with the necrosis observed in vivo. With hydroxyapatite, phagocytic activity of pulpal fibroblasts toward hydroxyapatite particles (< 10 microns) was seen. As a consequence, DNA synthesis was affected. This inhibitory effect was not due to cell damage, as demonstrated by increased [3H]-proline and [3H]-leucine incorporation by the cells. There was also an inhibitory effect of hydroxyapatite on alkaline phosphatase activity, suggesting that the pulp fibroblasts were not in a differentiation stage. In conclusion, compared to the effects of Ca(OH)2 on human pulp fibroblasts, these data are consistent with the biocompatibility of hydroxyapatite previously described in vivo and testify to the occurrence of a biological response elicited by this synthetic biomaterial.

Adolescent↗

PubMed1994

Rat liver cytochrome P450 2B3: structure of the CYP2B3 gene and immunological identification of a constitutive P450 2B3-like protein in rat liver.

The cytochrome P450 2B subfamily in the rat contains an estimated eight to eleven members at the genomic level. Synthesis in the liver of the prototypic forms P450 2B1 and P450 2B2 is dramatically induced by phenobarbital. The 1.9-kb mRNA for P450 2B3, a third member of the P450 2B subfamily, is constitutively present in rat liver but is not inducible by phenobarbital. We have now cloned and sequenced exonic sequences corresponding to the entire 2B3 mRNA and determined their exon-intron structure, which is identical to that of CYP2B1/CYP2B2 and other CYP2B genes. A putative CYP2B3 transcription start site was identified and CYP2B3 5'- and 3'-flanking sequences were compared to those of CYP2B1 and CYP2B2. CYP2B3, like CYP2B1 and CYP2B2, has a modified TATA box preceding the transcription start site and lacks the canonical polyadenylation signal preceding the poly(A) site. A 2B3 expression vector, pMT2-2B3, directed the synthesis in COS-1 cells of an approximately 50-kD protein detectable on Western blots with a polyclonal antibody and with one of four monoclonal antibodies raised against 2B1 but not with a polyclonal antibody raised against P450 PB6. The 2B3 protein migrated with a slightly higher electrophoretic mobility than 2B1 and comigrated with a protein detected by anti-2B1 antibodies in liver microsomes from untreated rats. The results indicate that a 2B3-like protein is present in rat liver and that it is distinct from P450 PB6 and other known constitutive rat hepatic P450s.

Animals↗

PubMed1994

Development of a monoclonal antibody against dentin phosphophoryn: a tool to study odontoblastic activity.

The role played by phosphophoryn, one of the major noncollagenous proteins of dentin extracellular matrix, in the mineralization process has not been fully characterized. The purpose of our work was to produce monoclonal antibodies (MAbs) against dentin phosphophoryn and to test their reactivity with primary culture of odontoblasts. Dentin phosphophoryn (DPP) was extracted after the mechanical dissociation of teeth and dialyzed against guanidine and EDTA solutions followed by CaC1(2) precipitation. These extracts were characterized by SDS-PAGE and staining with Coomassie blue and Stains-All. After immunization of mice with these extracts, we produced MAb 7G4, which reacted with dentin phosphophoryn as revealed by Western blot. MAb 7G4 reactivity was tested against a primary culture of pig odontoblasts, revealing filaments specifically stained by the anti-DPP antibody. This antibody will be of great interest to study the mineralization process and dental pulp reaction after capping with various calcium phosphate materials.

Animals↗

PubMed1993

Ionic basis for endogenous rhythmic patterns induced by activation of N-methyl-D-aspartate receptors in neurons of the rat nucleus tractus solitarii.

1. Activation of N-methyl-D-aspartate (NMDA) receptors in caudal nucleus tractus solitarii (cNTS) neurons elicited endogenous rhythmic activities. We used an in vitro brain stem slice preparation to determine the ionic mechanisms underlying the generation of these activities. 2. Using intracellular recordings, we found several ionic conductances to be responsible for the electrophysiological properties of cNTS neurons. After addition of tetrodotoxin (TTX) to the perfusate, cNTS neurons were still able to generate action potentials (APs). Because these APs were suppressed by the addition of cobalt or by the reduction of calcium, they were likely due to calcium currents (ICa). In addition, the amplitude of the afterhyperpolarization (AHP) that followed a train of TTX-resistant APs was reduced in both low-calcium and cobalt-containing saline. It was therefore suggested that calcium-activated potassium (IKCa) currents were involved in the AHP. Accordingly, application of apamin, a blocker of slow IKCa, also decreased the AHP. cNTS neurons exhibited a delayed excitation phenomenon, characterized by a ramplike depolarization that delayed the onset of neuronal firing, when they were depolarized from hyperpolarizing potential. The underlying current was presumed to be an A-current (IKA), because this phenomenon was suppressed during application of 4-aminopyridine (4-AP). 3. Application of NMDA elicited different types of discharge patterns in cNTS neurons: a repetitive firing at depolarized levels of membrane potential (above -60 mV) and rhythmic patterns characterized by either rhythmic bursting or rhythmic single discharges at hyperpolarized levels (within membrane potential range of -60 to -85 mV). In all neurons, rhythmic patterns were superimposed on oscillations of membrane potential. They were characterized by a sudden shift of membrane potential, followed by a ramp-shaped phase of depolarization that preceded spike elicitation. Addition of TTX to the saline did not suppress NMDA-induced oscillations. Therefore rhythmic patterns were not driven by synaptic mechanisms but resulted from endogenous properties of cNTS neurons. 4. APs superimposed on NMDA-induced depolarizations presented the same characteristics as those elicited by positive current pulses. NMDA-elicited oscillations of membrane potential were eliminated by removing magnesium from the saline. Therefore oscillation generation was based primarily on the NMDA channel properties. 5. Intrinsic conductances of cNTS neurons interacted with NMDA-gated conductances to shape the depolarization waveform. Because removal of calcium from the saline suppressed endogenous oscillations, ICa currents were required for the expression of rhythmic activities. IKCa currents were involved in the repolarization phase of oscillations because apamin increased the duration of the oscillations.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

PubMed1991

Evidence that activation of N-methyl-D-aspartate (NMDA) and non-NMDA receptors within the nucleus tractus solitarii triggers swallowing.

Swallowing is a patterned motor activity generated by neurons located within the nucleus tractus solitarii (NTS). Previous experiments have shown that administration of excitatory amino acids within the NTS induces swallowing. The present study was undertaken to identify the receptor subtypes involved in this effect. Pressure microinjections of L-glutamate (10-100 pmol), quisqualate (0.1-10 pmol) and N-methyl-D-aspartate (NMDA, 0.1-10 pmol) were performed into the NTS of decerebrate rats. Glutamate and quisqualate microinjections elicited short series of swallows while NMDA microinjections induced long-lasting, rhythmic swallowing. Pretreatment with the selective NMDA antagonist, DL-2-amino-5-phosphonovalerate (50 pmol), almost completely suppressed the response elicited by NMDA (10 pmol) but did not induce a significant modification of swallowing triggered by either glutamate (25 pmol) or quisqualate (10 pmol). Pretreatment with 6-cyano-7-nitroquinoxaline-2,3-dione (50 pmol), a selective blocker of non-NMDA receptors, suppressed the swallows elicited by glutamate and strongly inhibited the response elicited by quisqualate microinjections. The same pretreatment induced only a slight modification of the swallowing elicited by NMDA. These data demonstrate that deglutition can be triggered by activating either NMDA or non-NMDA receptors localized within the NTS, and therefore suggest that both receptor subtypes may be involved in swallowing elicited under physiological conditions.

2-Amino-5-phosphonovalerate↗

PubMed1991

Bursting discharges evoked in vitro, by solitary tract stimulation or application of N-methyl-D-aspartate, in neurons of the rat nucleus tractus solitarii.

Extracellular recordings of the activity of neurons in the isolated nucleus tractus solitarii (NTS) showed that repetitive stimulation of the tractus solitarius (TS) elicited either long-lasting discharges at low frequency (type A neurons) or short-duration bursting discharges at low or high frequency (type B and C neurons, respectively). Bath application of N-methyl-D-aspartate (NMDA; 60-120 microM; 4-5 min duration) elicited a pattern of rhythmic bursting in most type B (21/24) and in all type C neurons and only a repetitive firing in type A neurons, even with applications of longer duration (6-10 min). The rhythmic bursting pattern was characterized by trains of action potentials occurring at a regular rate with each neuron (mean = 0.9 +/- 0.45 Hz). The present findings suggest that local mechanisms within the NTS (synaptic interactions or neuronal intrinsic properties) are involved in the burst firing elicited either by TS stimulation or NMDA application. The data are discussed in terms of the generation of swallowing taking into account the key role of NTS neurons in the organization of this motor pattern.

Action Potentials↗

PubMed1991

Hospital ethics committees in Quebec: an overview.

In June 1989, La Direction de la Sante physique (ministere de la Sante et des Services sociaux) (MSSS) undertook a major research project focusing on ethics committees in Quebec, in cooperation with Laval University's Groupe de recherche en Ethique Medicale (GREM). Initially, three major objectives were set out: to prepare a faithful overview of ethics committees in Quebec; to inform all those involved of the available resources, and to collect the data required to eventually carry out in-depth research on ethics committees. A number of government documents refer to the existence of ethics committees, whether to request their cooperation or to entrust mandates to them; moreover, the Law Reform Commission of Canada recently published a document that proposed establishing a Canadian Advisory Council on Biomedical Ethics. The MSSS, therefore, had to study the situation in Quebec with regard to ethics committees in order to be in a position to clarify its position in future discussions. This overview is the outcome of the first stage of this research project. Essentially, it represents the first of three main sections of a single document. It [1] presents general data on ethics committees, [2] contains a description of each committee by socio-sanitary region and type of committee, and in section three it deals with the principal resources available in bioethics in Quebec, Canada, and throughout the world.

Clinical Protocols↗

PubMed1991

Scanning electron microscopic study of odontoblasts and circumpulpal dentin in a human tooth.

Two combined scanning electron microscopic methods including modified fixative procedures were used for studying the morphologic aspects of the odontoblasts and the corresponding dentinal wall at different endodontic levels. The odontoblasts were tightly packed in the pulp horn, where they assumed pear-shaped profiles and, from crown to apex, successively looked spindle shaped, club shaped, and globular. Their number decreased in the pulp radicular portion, where the interglobular spaces were enlarged. The filling fibrillar material varied from crown to apex. A globular circumpulpal dentin was observed at all levels of the root canal. The tubule openings varied in number and size.

Adult↗

PubMed1991

[The nucleus tractus solitarius: neuroanatomic, neurochemical and functional aspects].

The nucleus tractus solitarii (NTS) has long been considered as the first central relay for gustatory and visceral afferent informations only. However, data obtained during the past ten years, with neuroanatomical, biochemical and electrophysiological techniques, clearly demonstrate that the NTS is a structure with a high degree of complexity, which plays, at the medullary level, a key role in several integrative processes. The NTS, located in the dorsomedial medulla, is a structure of small size containing a limited number of neurons scattered in a more or less dense fibrillar plexus. The distribution and the organization of both the cells and the fibrillar network are not homogeneous within the nucleus and the NTS has been divided cytoarchitectonically into various subnuclei, which are partly correlated with the areas of projection of peripheral afferent endings. At the ultrastructural level, the NTS shows several complex synaptic arrangements in form of glomeruli. These arrangements provide morphological substrates for complex mechanisms of intercellular communication within the NTS. The NTS is not only the site of vagal and glossopharyngeal afferent projections, it receives also endings from facial and trigeminal nerves as well as from some renal afferents. Gustatory and somatic afferents from the oropharyngeal region project with a crude somatotopy within the rostral part of the NTS and visceral afferents from cardiovascular, digestive, respiratory and renal systems terminate viscero-topically within its caudal part. Moreover the NTS is extensively connected with several central structures. It projects directly to multiple brain regions by means of short connections to bulbo-ponto-mesencephalic structures (parabrachial nucleus, motor nuclei of several cranial nerves, ventro-lateral reticular formation, raphe nuclei...) and long connections to the spinal cord and diencephalic and telencephalic structures, in particular the hypothalamus and some limbic structures. The NTS is also the recipient of several central afferent inputs. It is worth to note that most of the structures that receive a direct projection from the NTS project back to the nucleus. Direct projections from the cerebral cortex to the NTS have also been identified. These extensive connections indicate that the NTS is a key structure for autonomic and neuroendocrine functions as well as for integration of somatic and autonomic responses in certain behaviors. The NTS contains a great diversity of neuroactive substances. Indeed, most of the substances identified within the central nervous system have also been detected in the NTS and may act, at this level, as classical transmitters and/or neuromodulators.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗

PubMed1990

Rhythmic bursting patterns induced in neurons of the rat nucleus tractus solitarii, in vitro, in response to N-methyl-D-aspartate.

The activity of nucleus tractus solitarii (NTS) neurons was recorded extracellularly on rat brainstem slices. Depending on the neuron, bath application of N-methyl-D-aspartate (NMDA; 30-120 microM) elicited either a pattern of rhythmic bursting or repetitive firing. The rhythmic bursting pattern was characterized by trains of action potentials occurring at a rate of 0.36-2 Hz. With most of the neurons, the mean burst duration and the mean discharge frequency ranged between 200 and 800 ms and between 20 and 40 Hz, respectively. Both the repetitive and the rhythmic bursting patterns were reversibly blocked when DL-2-amino-5-phosphonovalerate (80 microM) was applied. Application of quisqualate (30-60 microM) or glutamate (300-1200 microM) on NTS neurons induced only repetitive firing even in neurons exhibiting a rhythmic bursting pattern under NMDA. The present findings show that rhythmic bursting patterns can be generated within the isolated NTS under activation of NMDA receptors. The rhythmic bursting resulted probably from local NTS mechanisms (synaptic interactions or neuronal intrinsic properties) which might be involved in physiological rhythmic activities organized at the NTS level, such as swallowing.

2-Amino-5-phosphonovalerate↗