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Biomedical subjects

G Poirier

Publications and source records attributed to G Poirier.

At least 37 records · Page 2Linked to original sources
PubMed1990

Expression in E. coli of the catalytic domain of rat poly(ADP-ribose)polymerase.

A 2 kilobase pair cDNA coding for the entire C-terminal catalytic domain of rat poly(ADP-ribose)polymerase has been expressed in E. coli. The overproduced 55 kDa polypeptide is active in synthesizing poly(ADP-ribose) and the 4 kDa N-terminal region of this domain is recognized by the monoclonal antibody C I,2 directed against the calf enzyme. Also, the minor alpha-chymotrypsin cleavage site found in the human catalytic domain is not present in the rat enzyme as revealed by the absence of the 40 kDa specific degradation product in the E. coli cells expressing the rat domain. The expression of this partial rat cDNA should thus permit the rapid purification and subsequent crystallization of the catalytic domain of the enzyme.

Amino Acid Sequence↗
PubMed1990

Stimulation of poly(ADP-ribose) synthesis by free radicals in C3H10T1/2 cells: relationship with NAD metabolism and DNA breakage.

We have studied the effect of H2O2 and O2- produced by xanthine and xanthine oxidase on NAD catabolism, poly(ADP-ribose) synthesis, and production of DNA single-strand breaks in C3H10T1/2 cells. The results show a correlation between the induction of DNA single-strand breaks, the decrease of NAD pool, and the accumulation of polymer. New techniques, based on affinity chromatography and reversed-phase high pressure liquid chromatography, have allowed an accurate determination of polymer contents and showed a 20-fold stimulation of polymer biosynthesis induced by active oxygen species. Inhibition experiments performed with 3-aminobenzamide have shown that the decrease in NAD levels after exposure of cells to active oxygen species was caused by stimulation of poly(ADP-ribosyl)ation and of another cellular process.

Alkaline Phosphatase↗
PubMed1989

Poly(ADP-ribose)polymerase: a novel finger protein.

By Energy Dispersive X-ray fluorescence we have determined that calf thymus poly(ADP-ribose) polymerase binds two zinc ions per enzyme molecule. Using 65Zn (II) for detection of zinc binding proteins and polypeptides on western blots, we found that the zinc binding sites are localized in a 29 kd N-terminal fragment which is included in the DNA binding domain. Metal depletion and restoration experiments proved that zinc is essential for the binding of this fragment to DNA as tested by Southwestern assay. These results correlate with the existence of two putative zinc finger motifs present in the N-terminal part of the human enzyme. Poly(ADP-ribose)polymerase fingers could be involved in the recognition of DNA strand breaks and therefore in enzyme activation.

Amino Acid Sequence↗
PubMed1989

Anti-CD45RA antibodies increase the proliferation of mouse T cells to phytohemagglutinin through the interleukin 2/interleukin 2 receptor pathway.

We have studied the effect of anti-CD45RA monoclonal antibodies on the proliferation of murine spleen T cells activated with phytohemagglutinin. Antibodies act on both CD8 and CD4 subsets of T cells. They seem to replace a signal normally delivered by accessory cells which would act through the interleukin 2/interleukin 2 receptor pathway.

Animals↗
PubMed1989

Differences in the release of 5'-nucleotidase and alkaline phosphatase from plasma membrane of several cell types by PI-PLC.

1. We have compared the effect of phosphatidyl inositol specific phospholipase C (PI-PLC) on the attachment of both 5'-nucleotidase and alkaline phosphatase to the liver plasma membrane from different species. 2. Our results demonstrate differences in the susceptibilities of both enzymes to PI-PLC treatment in relation to their origin. 3. These results were confirmed by immunoblotting using polyclonal anti-5'-nucleotidase antibodies. 4. In addition, in a single animal, susceptibility of both enzymes to PI-PLC treatment is different from one tissue to another. 5. The different percentages of released enzymes could be explained either by a polymorphism in the anchoring of these proteins at the cell surface membrane, or by a different steric hindrance or environment at the cleavage site itself.

5'-Nucleotidase↗
PubMed1989

The effects of L-dopa on periodic leg movements and sleep organization in narcolepsy.

A large proportion of narcoleptic patients have periodic leg movements (PMS) in sleep. The contribution of these movements to the nocturnal sleep disturbance observed in narcoleptics remains a controversial issue. The aim of the present study was to look at the sleep organization of narcoleptic patients before and after suppression of periodic leg movements with L-dopa. L-dopa and a placebo were administered in a double-blind fashion to six narcoleptic patients. Each treatment period lasted 2 weeks and the treatment order was reversed for one-half of the subjects. The effects of L-dopa and placebo were evaluated by polysomnography. A significant reduction of PMS was seen after treatment with L-dopa, but this treatment did not improve sleep organization. On the contrary, L-dopa increased wake time after sleep onset. This result supports the hypothesis that PMS does not play a major role in the nocturnal sleep disruption observed in narcolepsy. It also supports the hypothesis that dopaminergic mechanisms play a role in the physiopathology of PMS.

Adolescent↗
PubMed1989

Problems in the physiology of class I and class II MHC molecules, and of CD45.

1. Co-processing of alloantigens suggests that epitope-loaded MHC class I molecules may pass from tissue cells to dendritic cells. 2. Antigen-presenting cells in the thymus need some special trick in order to load their MHC class II molecules with epitopes from "intermediate concentration" self-proteins in order to induce self-tolerance in developing cells. 3. Cell-cell interactions may transmit signals simply by rearranging surface glycoproteins and thus locally perturbing a phosphorylation equilibrium. 4. The CD45 and STB1 phenotype of most cells in the thymus may be characteristic of a doomed cell.

Animals↗
PubMed1987

Clinical and sleep laboratory study of narcoleptic symptoms in multiple sclerosis.

Seventy white patients with a diagnosis of MS and typed for their HLA-A, B, C, and DR were studied. A clinical interview and a questionnaire were used to evaluate the presence of narcoleptic symptoms. The prevalence of sleep attacks, cataplexy, and sleep paralysis was significantly elevated among these patients. However, no difference was seen between DR2 and non-DR2 subjects with regard to the incidence of narcoleptic symptoms. Nine DR2 patients complaining of both sleep attacks and cataplexy were studied in the sleep laboratory for five consecutive naps, but no polygraphic evidence of narcolepsy was found.

Adult↗
PubMed1986

HLA antigens in narcolepsy and idiopathic central nervous system hypersomnolence.

Several studies have shown familial incidence of narcolepsy and idiopathic central nervous system (CNS) hypersomnia. HLA antigen studies performed in mongoloid and caucasoid narcoleptic patients on the A, B, and C loci have yielded conflicting results. The aim of this study is to document a possible association between the HLA system, including the DR locus and excessive daytime somnolence. Thirty-one narcoleptic patients and 10 idiopathic hypersomniac patients were selected and typed for 54 HLA antigens. A family with narcoleptic members in 3 generations was also studied. HLA-DR2 was found in 100% of narcoleptic patients. The frequency of HLA-A3 and B7, which are in linkage disequilibrium with DR2 was also increased in this group. Idiopathic hypersomniac patients showed an increase of HLA-Cw2, DR5, and B27, three antigens known to be in linkage disequilibrium. In the family study, narcoleptic patients were also HLA-DR2; moreover, 3 subjects, one of whom was narcoleptic, were HLA-DR2 as a result of recombination (i.e., genetic crossing-over). These results locate the hypothetic gene associated with narcolepsy more precisely, and indicate that narcolepsy and idiopathic CNS hypersomnolence are two different entities.

Adult↗
PubMed1986

Viloxazine hydrochloride in narcolepsy: a preliminary report.

Twenty-three narcoleptic subjects participated in a single-blind drug study with placebo-viloxazine hydrochloride (100 mg/day). One woman discontinued the study because of nausea and headaches during the treatment period. The remaining 22 subjects were polygraphically monitored during baseline, The remaining 22 subjects were polygraphically monitored during baseline, placebo intake, viloxazine treatment, and placebo intake following 48 h of drug withdrawal. Symptoms and side effects were evaluated subjectively and objectively. Except for the subject who discontinued the study, viloxazine was well tolerated, particularly in elderly subjects. The drug had an inhibitory effect on REM sleep, cataplexy, and other auxiliary symptoms. While there is no objective evidence that it heightens alertness, patients reported fewer sleep attacks.

Adult↗
PubMed1986

Restless legs syndrome and periodic movements in sleep: physiopathology and treatment with L-dopa.

Seven patients suffering from restless legs syndrome (RLS) and periodic movements in sleep (PMS) were investigated before and after treatment with L-Dopa. The effect of treatment was evaluated by polysomnography, structured interviews, and daily questionnaires. Sleep organization and subjective complaints improved during treatment with 100 to 200 mg of L-Dopa. Polysomnographic recordings also revealed a significant decrease of periodic leg movements during the first third of the night and a rebound during the last third. These results and previous biochemical findings raise the hypothesis that RLS and PMS may both result from reduced dopaminergic activity in the CNS, perhaps resulting from decreased sensibility of postsynaptic receptors.

Adult↗
PubMed1986

Production and characterization of monoclonal antibodies specific for the functional domains of poly(ADP-ribose) polymerase.

Monoclonal antibodies were developed against poly(ADP-ribose) polymerase and analyzed for their reactivity against the NAD+- and DNA-binding fragments. Two fusions were performed to obtain hybridomas and the resulting anti-poly(ADP-ribose) polymerase antibodies were further screened by characterization of their immunoglobulin light chains. Five different hybridomas were isolated which produced different immunoglobulin light chains, all of which were specific for poly(ADP-ribose) polymerase. The specificities of these antibodies were determined by immunoblotting against the purified poly(ADP-ribose) polymerase, its autodegradation fragments, and the fragments prepared by limited proteolysis with chymotrypsin and papain. These fragments have been suggested to contain the NAD+-binding site, the DNA-binding site, and the automodification site, respectively. All the monoclonal antibodies reacted with the 116 kdalton (kDa) band corresponding to the purified enzyme. Four antibodies reacted exclusively with antigenic site(s) on the 46-kDa fragment which contains the DNA-binding site. A fifth antibody reacted exclusively with a clearly different antigenic site on the 74- and 54-kDa fragments which possess the NAD+ (substrate) binding site. The immunoreactivity with the major autodegradation products (69- and 46-kDa fragments) of the purified enzyme confirms this difference between the two groups of antibodies. The 22-kDa fragment corresponding to the auto-modification site does not show any immunoreactivity with the antibodies.

Animals↗
PubMed1985

Modulation of rat pancreatic muscarinic cholinergic receptors by caerulein.

To evaluate the modulation of pancreatic muscarinic receptors in two states of pancreatic growth, hypertrophy and hyperplasia, caerulein, a cholecystokinin analog, (1 microgram/kg) was administered thrice daily for 2 and 4 days to adult rats. After 2 days of treatment, pancreatic hypertrophy was well established as evidenced by increases in pancreatic weight, cellular mass and protein content. Using an increase in DNA content as an index of hyperplasia, we demonstrated that pancreatic hyperplasia occurred only after 4 days of caerulein treatment. Caerulein increased the concentration of muscarinic receptors per DNA in pancreatic homogenate by 57% over control value after 2 days of treatment without modification of the receptor affinity for the ligand QNB. This increase involved mainly receptors in the low affinity state for carbamylcholine and their concentration returned to control levels after 4 days of treatment. The functional capacity of the acini was significantly increased after 2 days of caerulein as amylase release (U/mg DNA) was significantly increased but the sensitivity of these acini to carbamylcholine was significantly decreased. After 4 days of caerulein, the functional capacity has returned towards control values but the sensitivity to carbamylcholine remained decreased. The increase in muscarinic receptor concentration could be ascribed to a general increase in cellular proteins, as part of the hypertrophic effect of caerulein. This specific effect would also explain the increased functional secretory capacity of the caerulein-treated acini but the decreased sensitivity to carbamylcholine probably resulted in changes at a postreceptor loci since the affinities of the muscarinic receptors for carbamylcholine remained unaffected.

Amylases↗
PubMed1985

Tumor promoter phorbol-12-myristate-13-acetate induces poly ADP-ribosylation in human monocytes.

The tumor promoter phorbol-12-myristate-13-acetate (PMA) induces rapid poly ADP-ribosylation and a drop in cellular NAD concentration in human monocytes. The antioxidants CuZn-superoxide dismutase, catalase, glutathione peroxidase and butylated-hydroxytoluene inhibit the reaction indicating that active oxygen species produced in the PMA-induced oxidative burst represent intermediates. The inhibitor of ADP-ribosyl-transferase, 3-amino-benzamide, inhibited poly ADP-ribosylation but did not prevent the drop in NAD-levels. PMA also causes the slow accumulation of DNA strand breaks in monocytes. The difference in the kinetics of poly ADP-ribosylation and DNA breakage argues against a simple relationship between the two reactions.

Antioxidants↗
PubMed1985

Tumor promoter phorbol-12-myristate-13-acetate induces poly(ADP)-ribosylation in fibroblasts.

The tumor promoter phorbol-12-myristate-13-acetate (PMA) causes an increase in pol(ADP)-ribosylation in mouse and human fibroblasts via the intermediate formation of active oxygen. In contrast to poly(ADP)-ribosylation induced by the methylating agent N-methyl-N'-nitro-N-nitrosoguanidine, de novo RNA and protein synthesis are required and the accumulation of the polymer occurs in the absence of detectable DNA strand breakage. Our results suggest a mechanism for PMA-induced modulation of chromatin structure and gene expression.

Animals↗