Biomedical subjects

N Moreau

Publications and source records attributed to N Moreau.

At least 19 recordsLinked to original sources

PubMed1998

Young age and HLA markers enhance the risk of progression to type 1 diabetes in antibody-positive siblings of diabetic children.

The contribution of autoantibodies, HLA markers and age to long-term estimates of risk of type 1 diabetes were examined after a median of 11 years (range 7.5-14) during the follow-up in a cohort of 234 siblings (aged 2-29 years) of French children with recent-onset type 1 diabetes, of whom 12 (5.1%) developed diabetes. We evaluated islet cell antibodies (ICA) by indirect immunofluorescence and autoantibodies to insulin (IAA), to the 65 kDa isoform of glutamic acid decarboxylase (GADA) and to the IA-2 protein (IA-2A) by radioligand assay in sequential serum samples. Among the 234 siblings of type 1 diabetic patients screened, 27 were positive for at least one antibody, 11 of whom progressed to develop type 1 diabetes during the follow-up (sensitivity, 92%, predictive value, 41%). Among the four antibodies tested individually, ICA had the highest sensitivity (83%) but a poor predictive value (59%) and IA-2A the highest predictive value (70%). IAA and GADA both exhibited poor sensitivity and predictive value. Combinations of antibodies achieved better predictive values than antibodies tested individually. Satisfactory predictive values were obtained for the combination of GADA with IA-2A (83%), for any combination of at least two antibodies other than ICA (70%) and for the combination of ICA with at least one other antibody (69%). The risk estimates were highest in the presence of three or four antibodies, whether comprising ICA or not, but with a concomitant loss of sensitivity. For most antibody combinations, cumulative risks showed progression from approximately 50% after 5 years to 100% after 13 years. HLA-DR3/4 was significantly more frequent in siblings developing type 1 diabetes than in non-diabetic siblings (9/12 vs. 39/217, relative risk (RR)=14, P</=0.0001). The predictive value of HLA-DR3/4 was low (19%); however, taking into account the presence of HLA-DR3/4 in subjects who were positive for more than one antibody resulted in a higher predictive value (67%, vs. 20% in non-DR3/4 subjects, P</=0.02). In addition, siblings developing diabetes were younger at entry than those who did not (mean =7.5 +/-1.23 vs. 12.5 +/-0.39 years, respectively; P</=0.01). Ten of 12 were aged less than 10 years compared with 106/222 non-diabetic siblings (RR =5.4, P</=0.03). Moreover, younger age was associated with a more rapid development of type 1 diabetes. In conclusion, our results show that the combination of IAA, GADA and IA-2A autoantibodies in sequential serum samples is satisfactory for the identification of subjects at risk of developing type 1 diabetes. Additional factors such as younger age and HLA-DR3/4 as markers of progression to disease may contribute to more efficient prediction in antibody positive subjects.

Adolescent↗

PubMed1998

Evidence for specific RNA/protein interactions in the differential segment of the W sex chromosome in the amphibian Pleurodeles waltl.

Pleurodeles exhibits a ZZ/ZW system of GSD (genotype sex determination). However, the Z and W sex chromosomes appear to be morphologically identical. A short RNA sequence is described that was specifically bound to lampbrush loops in the differential segment of the sexual bivalent IV. The distribution of these labeled loops in experimentally produced ZZ and WW females enabled us to demonstrate that such labeled loops were perfectly correlated with the W chromosome. Therefore, this RNA sequence constitutes an excellent marker for the W differential segment. Furthermore, analysis of the labeled loops under various experimental conditions suggested that their labeling is caused by specific interactions between this RNA sequence and lampbrush loop-associated proteins (RNA/protein interactions). North-western assays revealed that nuclear polypeptide(s) of 65 kDa could be responsible for such binding.

Animals↗

PubMed1998

Cell-cycle-dependent nuclear translocation of HSP70 in amphibian embryonic cells.

Using immunocytochemical methods, we analyzed the localization of the HSP70 protein constitutively expressed during embryogenesis in the amphibian Pleurodeles waltl. Our results provide evidence for nuclear transfer of the protein during gastrulation, and particularly for predominant nuclear labeling in gastrula internalized cells. Using two inhibitors of DNA replication -hydroxyurea (HUA) and aphidicolin- or/and an inhibitor of transcription -actinomycin D- applied to embryos, we demonstrated that nuclear transfer of HSP70 is related to the transcriptional activity of the cells during the early S phase of the cell cycle.

Animals↗

PubMed1997

Conditions for a heat shock response during oogenesis and embryogenesis of the amphibian Pleurodeles waltl.

The optimal conditions capable of inducing an increase in HSP70 neosynthesis during development of the urodele amphibian Pleurodeles waltl were determined in this study. These conditions depend on temperature, heat shock duration and recovery duration. In oocytes, a heat shock response was repeatedly obtained at 37 degrees C for 15 min followed by 1 h recovery. These results provided evidence for heat shock response at every stage considered. An increase in HSP70 synthesis was noted throughout oogenesis, but it did not lead to an increase in the amount of soluble HSP70, except for stage VI oocytes. Such results suggest that from stage II to stage IV oocytes, an equilibrium occurs between the HSP70 used and the HSP70 neosynthesized. In contrast, in stage VI oocytes, heat shock led to overproduction of HSP70. During early development, the heat shock response was repeatedly obtained only from the gastrula stage with a 37 degrees C shock and a 15 min duration of treatment. Surprisingly, during cleavage stage, the soluble HSP70 total amount increased after heat shock at a time when no HSP70 neosynthesis occurred.

Amino Acid Sequence↗

PubMed1996

Sequence analysis, purification, and study of inhibition by 4-quinolones of the DNA gyrase from Mycobacterium smegmatis.

We determined the nucleotide sequence of a 6-kb DNA region harboring the recF, orf192, gyrB, and gyrA genes from Mycobacterium smegmatis mc(2)155. The amino acid sequences deduced from gyrA and gyrB displayed 89 and 86% identity, respectively, with the DNA gyrase from Mycobacterium tuberculosis, and 67 and 65% identity, respectively, with that from Streptomyces coelicolor. An open reading frame encoding the C-terminal region of the M. smegmatis RecF polypeptide was found upstream from gyrB and was 57% identical to the open reading frame encoding the C-terminal region of the S. coelicolor RecF protein. The gene orf192 was identified between recF and gyrB and was 39% identical to orf191 found in S. coelicolor in the recF-gyrB region. The M. smegmatis DNA gyrase, which was purified by affinity chromatography on novobiocin-Sepharose, consisted of two polypeptides with apparent molecular masses of 98 and 80 kDa. Determination of the N-terminal amino acid sequence of the B subunit confirmed GTG as the start codon in gyrB. Analysis of the supercoiling activity of the enzyme indicated that the M. smegmatis DNA gyrase was characterized by a specific activity equivalent to that of the Escherichia coli DNA gyrase. Inhibition of this activity by 4-quinolones was investigated by determining the 50% inhibitory concentrations (IC50S) of nalidixic acid, ofloxacin, and ciprofloxacin. The results indicated that the inhibitory activities of these drugs against the M. smegmatis DNA gyrase were markedly lower than those previously reported for the E. coli DNA gyrase. The results also suggested that the higher levels of activity of ofloxacin and ciprofloxacin against M. smegmatis (MICs, 0.5 to 1 microgram/ml), in contrast to that of nalidixic acid (MIC, 256 micrograms/ml), could be related to the higher inhibitory activities of fluoroquinolones against the DNA gyrase from this species (IC50S, 7 to 14 micrograms/ml) compared with that of nalidixic acid (IC50, 1,400 micrograms/ml).

4-Quinolones↗

PubMed1996

Does the chaperone heat shock protein hsp70 play a role in the control of developmental processes?

Expression of an hsp70 gene strictly inducible in somatic cells and constitutively expressed during oogenesis was investigated during embryogenesis of the amphibian Pleurodeles waltl. Results from Northern hybridization experiments and RNase protection assays provided evidence for the presence of inducible hsp70 mRNA under normal conditions at every embryonic stage. Immunoblotting of embryo proteins separated by 2D-electrophoresis provided evidence for the presence of a single polypeptide of about 74 kDa likely to be an HSP70-related protein, from unfertilized egg to tailbud stage. Immunocytological analysis showed that HSP70-related proteins were localized in the cytoplasm of all blastomeres. It also pointed out that nuclear transfer of the protein occurs in certain cells, precisely at the time of their invagination and subsequent internalization during normal Pleurodeles development. Such nuclear transfer involves involuting mesodermal cells in the blastopore region at the time of gastrulation. It also involves neurodermic cells at the time of neural tube closure. Interestingly, in exogastrulas nuclear transfer did not occur in cells which could no longer invaginate. Such behavior of HSP70-related proteins led us to suggest that they are involved in the control of nuclear activity associated with important developmental events such as cellular internalization processes. Such a role may be a direct consequence of HSP70-related protein functional properties as molecular chaperones.

Animals↗

PubMed1995

Evidence for a 90 kDa heat-shock protein gene expression in the amphibian oocyte.

In order to study expression of a 90-kDa heat-shock protein during amphibian oogenesis at physiological temperature, we isolated a Pleurodeles waltl hsc90 cDNA by screening an ovarian cDNA library with a chicken hsp90 cDNA probe. The cDNA thus obtained--named Pw90--shows a high homology level with the hsp90 gene in other species. RNase protection analysis led us to conclude that this sequence is part of the cognate gene hsc90 and is constitutively expressed in oocytes. Furthermore, results of quantitative Northern blot analysis, as well as in situ hybridizations on oocyte sections or lampbrush chromosome spreads, provide evidence for expression of hsc90 transcripts at every stage of oogenesis. Moreover, they point to the fact that an accumulation of transcripts occurs very early in oogenesis. Simultaneously, the expression of HSC90-related protein was analyzed on Western blots using a monoclonal antibody (AC88) and a polyclonal antibody (AP90Ct) raised against the Pleurodeles C-terminal part of HSC90. We provide evidence for a net accumulation of HSC90-related protein in oocytes. Immunolocalization shows that a nuclear transfer occurs in the course of oogenesis and leads to a concentration equilibrium between cytoplasm and nucleus in stage VI oocytes.

Amino Acid Sequence↗

PubMed1994

Transcription of amphibian lampbrush chromosomes is disturbed by microinjection of HSP70 monoclonal antibodies.

To investigate the possible involvement of HSP70 in nuclear transport of proteins associated with the transcription process in amphibian oocyte lampbrush chromosomes, we examined the effect of anti-HSP70 mouse monoclonal antibodies on the transcriptional activity of lampbrush chromosomes. When injected into the oocyte cytoplasm, anti-HSP70 induced disorganization followed by retraction of chromosomal lateral loops. This retraction reflected inhibition of transcription, which was confirmed by our electron microscopic observations. At the same time, while nucleoli were morphologically disorganized, nucleolar transcription persisted. These modifications were completely reversible. In contrast, no modifications were observed when antibodies were directly microinjected into nuclei. Similar observations were registered at the level of lampbrush chromosomes when wheat germ agglutinin was injected into oocyte cytoplasm. All of these results suggest that HSP70 mediates the nuclear transport of proteins involved in the lampbrush chromosome transcriptional process.

Animals↗

PubMed1994

Vascular responsiveness in young, diabetic, and aging hyperinsulinemic rats.

The purpose of this study was to compare vascular responsiveness in young (12 week old), aging hyperinsulinemic-glucose intolerant (52 weeks old) and diabetic (streptozotocin; 14 weeks old) rats. Aortic rings with and without endothelium were maintained in organ chambers for isometric tension recording. The contractile response to KCl was significantly enhanced in aortae from diabetic animals when compared to the responses obtained in young and old ones. The contractile response to norepinephrine or U46619, was significantly shifted to the right in the aortae from aging animals, however the aortae from these hyperinsulinemic rats were hyperresponsive to serotonin. Acetylcholine and ADP provoked an endothelium-dependent relaxation which was markedly depressed in the aortae from diabetic animals. The relaxation to ADP was selectively inhibited in the aging animals. The effect of sodium-nitroprusside was not significantly different in the three groups. Isoproterenol and forskolin induced endothelium-independent relaxation. Isoproterenol responses were inhibited in aging and diabetic animals, however the forskolin-relaxation was inhibited only in the aortae from aging animals. These results suggest that in two models of diabetes (i.e. Type I insulin-dependent and type II non insulin-dependent) vascular responsiveness is differently affected. Aging hyperinsulinemic animals present a selective hyperresponsiveness to serotonin, a selective dysfunction of ADP-induced endothelium-dependent relaxation and smooth muscle adenylate cyclase deficit. In diabetic animals a beta adrenergic hyporesponsiveness, not linked to adenylate-cyclase dysfunction, and non-selective depression of endothelium-dependent responses can be observed.

Age Factors↗

PubMed1994

[Current status of resistance of bacteria to pefloxacin in hospital units. Cross resistance with ofloxacin and ciprofloxacin. Results of a multicenter study].

The antibacterial activities of pefloxacin (PEF), ofloxacin (OFL), ciprofloxacin (CIP) were measured by agar diffusion (5 micrograms strength disks) against 6370 non duplicate clinical isolates collected in November 1991 in 40 hospitals from various areas in France. The MICs of PEF, OFL and CIP were determined by agar dilution against intermediate or resistant strains to one of the three antibiotics (inhibition zone < 16 mm for PEF, OFL, < 19 mm for CIP). In the Enterobacteriaceae the overall incidence of resistance to PEF (MIC > 4 mg/l) was 8% with important variations between the different species: E. coli 2%, Salmonella 0%, E. cloacae 9%, K. pneumoniae 21%, P. mirabilis 13%, P. vulgaris 3%, M. morganii 5%, Providencia 61%, S. marcescens 55%. Among S. aureus, the incidence of resistance was 3% for the methicillin-susceptible strains and 86% for the methicillin-resistant strains. The same applied to coagulase negative Staphylococci: methicillin-susceptible 9%, methicillin-resistant 62%. The frequency of the resistant strains was high among P. aeruginosa: 40% and A. baumanii: 83%. A high degree of correlation was observed between the MICs of PEF, OFL and CIP for all the bacterial species: r = 0.8 - 0.9. No major discrepancies were noted between the clinical categorizations of the activities of the three antibiotics for the different species except for P. aeruginosa: 4% of the strains were resistant to pefloxacin and susceptible to ciprofloxacin.

Ciprofloxacin↗

PubMed1993

Influence of sub-inhibitory concentrations of antibacterials on the surface properties and adhesion of Escherichia coli.

The effect of sub-inhibitory concentrations of antibacterials, including quinolones, on the surface properties of a uropathogenic strain of Escherichia coli was examined. The effect on the charge and hydrophobicity of the cell surface was assessed by means of partition between two aqueous phases, polyethylene glycol and dextran. Antibiotics at 1/8 x MIC inhibited adhesion to uroepithelial cells, and induced an increase in bacterial charge and hydrophobicity. Inhibition of adhesion correlated with increased charge, but not with hydrophobicity. The influence of magnesium on the inhibition of adhesion by sub-MICs of pefloxacin was also investigated. Loss of the anti-adhesive property of pefloxacin was observed with increasing magnesium concentrations, suggesting that quinolones should be free from magnesium to induce an inhibition of adhesion. Examination by electron microscopy showed a disappearance of fimbriae following treatment of E. coli cells with 1/8 x MIC of pefloxacin.

Anti-Bacterial Agents↗

PubMed1993

Effect of aging and drug-induced weight reduction on rat vascular reactivity.

We determined the effects of D-fenfluramine treatment on the changes in vascular reactivity induced by aging. Nine- and 49-week-old Sprague-Dawley rats (a strain known to develop hyperinsulinemia and glucose intolerance during the aging process) were treated for 3 weeks either with D-fenfluramine 2.5 mg/kg twice daily orally or with vehicle. The rats were then exsanguinated and the abdominal aorta was carefully removed, cut into rings, and suspended in organ chambers for isometric tension recording. Control old rats (vehicle) had a significantly lower glucose infusion rate (an index of insulin resistance), and higher blood pressure (BP), glycemia, and insulinemia than young rats. The D-fenfluramine treatment in the aged animals produced a significant decrease in insulinemia and body weight. In aorta from the older treated and nontreated animals, the contraction to alpha-adrenergic stimulation and to the thromboxane analogue U46619 was significantly reduced as compared with that in young animals, but the response to KCl was unaffected. In contrast, in the old nontreated rats, the aorta was hyperresponsive to serotonin. D-Fenfluramine abolished this hyperreactivity. The response to beta-adrenergic stimulation and to forskolin was inhibited in the older animals but was not influenced by the treatment. Endothelium-dependent relaxations to acetylcholine were not statistically different in the various groups, but the endothelium-dependent relaxation to ADP was reduced in the control group of older animals. D-Fenfluramine treatment restored the response to ADP.

Administration, Oral↗

PubMed1993

Sympathoinhibitory effects of losartan in spontaneously hypertensive rats.

Nonselective inhibition of endogenous angiotensin II (AII) by AI-converting enzyme inhibitors (ACEI) results in sympathoinhibitory effects. We wished to examine the influence of selective inhibition of endogenous AII by losartan, a nonpeptide AT1-receptor antagonist, on the sympathetic system. Cardiac, systemic, and regional vascular (kidney, mesentery, hindlimb) responses to selective alpha 1- and alpha 2-adrenoceptor agonists and to electrical stimulation of the spinal cord were investigated in pithed spontaneously hypertensive rats (SHR) by pulsed Doppler technique. Losartan (10 mg/kg) was administered orally, either as a single dose or for 8 successive days. Under both conditions, AII systemic pressor, regional vasoconstrictor, and tachycardic responses were completely abolished by losartan. At the vascular level, losartan did not affect postsynaptic alpha 1-adrenoceptor-mediated systemic pressor and regional vasconstrictor responses, but reduced postsynaptic alpha 2-adrenoceptor-mediated renal vasoconstriction. Losartan significantly decreased the systemic pressor and regional vasoconstrictor responses elicited by spinal cord stimulation. This sympathoinhibitory effect was not homogeneously distributed, preferentially affecting the kidney. At the cardiac level, spinal cord stimulation induced a strong tachycardia which remained unaffected by losartan. Thus in SHR, losartan exerts sympathoinhibitory effects against the vascular but not the cardiac responses to spinal cord stimulation. Because the vascular responses to postjunctional alpha 1- and alpha 2-adrenoceptor stimulation, except in the kidney, simultaneously remain poorly affected, the sympathoinhibitory effects of losartan mainly develop prejunctionally through AT1-receptors blockade.

Adrenergic alpha-Agonists↗

PubMed1993

Constitutive expression of a somatic heat-inducible hsp70 gene during amphibian oogenesis.

We isolated and characterized a sequence coding for heat-shock protein 70 (HSP70) of the amphibian Pleurodeles waltl. Results from S1 nuclease protection assays led us to conclude that an hsp70 gene, strictly inducible in somatic cells during heat shock, is constitutively active during oogenesis. By quantitative northern and western blot analysis, we showed that both hsp70 mRNA and HSP70-related protein levels increased in oocytes from stage II to stage VI under physiological conditions. Furthermore, by in situ hybridization to the nascent transcripts of lampbrush chromosome loops, we provided evidence for a clear-cut relationship between this increase in hsp70 mRNA and transcriptional activity during the lampbrush stage of oogenesis. These results strongly suggest that hsp70 genes are actively transcribed throughout oogenesis. HSP70-related proteins localized in the cytoplasm of young oocytes are progressively transferred to the nucleus in the course of oogenesis and preferentially accumulated in the nuclei of some stage VI oocytes.

Amino Acid Sequence↗

PubMed1993

[Interaction between SR 47436, a new angiotensin II antagonist and sympathetic nervous system in pithed SHR rats].

In vivo studies have previously shown that exogenous angiotensin II (AII) reinforces sympathetic nervous system activity. Conversely, non selective inhibition of endogenous AII by angiotensin I converting enzyme inhibitors (ACEIs) results in sympathoinhibitory effects. The aim of the present study was to examine the influence of selective inhibition of endogenous AII by SR 47436, a non peptide AT1-receptor antagonist, on the sympathetic nervous system. Cardiac, systemic and regional vascular (kidney, mesentery, hindlimb) responses to selective alpha 1- and alpha 2-adrenoceptor agonists and to electrical stimulation of the spinal cord were investigated in the pithed spontaneously hypertensive rat (SHR). Male adults SHRs were orally treated by SR 47436 (10 mg/kg/day for 8 days) or by distilled water. Two hours later, they were anesthetized with pentobarbital (50 mg/kg, i.p.), pithed and artificially ventilated. Blood pressure, heart rate, cardiac output and regional (kidney, mesentery and hindlimb) blood flows (pulsed Doppler technique) were measured. Corresponding vascular resistances were calculated. Three hours after SR 47436--at the time of the drug's maximal effects--or distilled water administration, cardiac, systemic pressor and regional vasoconstrictor responses (a) to increasing i.v. doses of AII, (b) to increasing frequencies of electrical stimulation of the spinal cord, and (c) to increasing i.v. doses of cirazoline, a selective alpha 1-adrenoceptor agonist, and of UK-14,304, a selective alpha 2-adrenoceptor agonist, were investigated. AII systemic pressor, regional vasoconstrictor and tachycardic responses were completely abolished by SR 47436. SR 47436 significantly reduced the systemic pressor responses elicited by spinal cord stimulation, cirazoline and UK-14,304.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗

PubMed1993

Immunolocalization of HSP 70-related proteins constitutively expressed during Xenopus laevis oogenesis and development.

Using immunocytochemical and biochemical methods, we analyzed the localization of HSP 70-related proteins constitutively expressed during oogenesis and embryogenesis in the amphibian Xenopus laevis. Our results provided evidence for a regional localization in oocytes. In embryos, the regional distribution observed in oocytes was found to be maintained from fertilization up to late blastula. It is noteworthy that, at the beginning of gastrulation, nuclear transfer of such proteins had already occurred by the time of internalization in the involuting marginal zone (IMZ), whereas cells of the vegetal area exhibited only a perinuclear localization of these proteins. These results suggest that HSP 70-related proteins might be involved in the control of the process of cellular internalization.

Animals↗

PubMed1992

Relationships among antibacterial activity, inhibition of DNA gyrase, and intracellular accumulation of 11 fluoroquinolones.

A series of 11 fluoroquinolone antibacterial agents, including 8 newly synthesized molecules and 3 reference compounds (pefloxacin, ciprofloxacin, and sparfloxacin), were tested for their MICs against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. The intracellular accumulation of fluoroquinolones by these microorganisms was measured by centrifugation through silicone oil and a fluorescence assay. The minimal effective dose (MED) was determined for all agents in a supercoiling assay with E. coli DNA gyrase. The hydrophobicities of the quinolones were determined and expressed as the logarithm of the coefficient of distribution (log D) between 1-octanol and phosphate buffer (pH 7.2). No correlation was found between MICs and cell accumulation for the quinolones studied. A correlation was found between log D and accumulation by S. aureus (r = 0.71, n = 11), and an inverse correlation was found between log D and accumulation by E. coli (r = 0.73, n = 11) and P. aeruginosa (r = 0.64, n = 10). The correlation coefficients between MICs and MED for E. coli, which were 0.60, 0.64, and 0.74 (n = 11) for E. coli, P. aeruginosa, and S. aureus, respectively, rose to 0.85, 0.74, and 0.74 (n = 11) for the same microorganisms, respectively, when the accumulation of the drug by the cell was taken into account. It was concluded that the inhibitory activity against DNA gyrase remains the most important parameter for quinolone potency, but that intracellular accumulation must be taken into account, since, for a given organism, both parameters are under the control of the physicochemical properties of the quinolones.

Anti-Infective Agents↗

PubMed1991

Comparison between in vivo and in vitro heat-induced changes in amphibian lampbrush chromosomes.

At normal breeding temperature (20 degrees C), amphibian lampbrush chromosomes are characterized by the presence of lateral loops which are related to the transcriptional process. Heat treatment induces changes in these loops, but the nature and timing of these modifications depend on hyperthermic stress conditions. Indeed, our data demonstrate that, at the same high temperature (34 degrees C), lampbrush chromosome modifications induced by in vivo and in vitro gradual heat treatments are different from those induced by in vitro heat shock. In vivo and in vitro heat treatments lead to progressive disorganization of landmark loops, whereas in vitro heat shock results in chromosome condensation. The progressive adaptation of lampbrush chromosome structure in response to gradual heat stress is considered and discussed.

Animals↗