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G Beauregard

Publications and source records attributed to G Beauregard.

At least 37 records · Page 2Linked to original sources
PubMed1987

Radiation inactivation of hamster acrosin reveals that the biologically active unit is of low molecular size.

The relationship between structure and activity of acid-extracted and purified acrosin obtained from cauda epididymal hamster spermatozoa was studied. A four-step purification procedure of acrosin was used; it included 1.) acid extraction, 2.) gel filtration over Sephadex G-100 resin, 3.) ion exchange on CM-Sepharose CL-6B, and 4.) affinity chromatography on proflavin-Sepharose 4B. Analysis of the purified enzyme by high-performance liquid chromatography (300 SW + I-125) revealed a molecular weight of 44,000, which was identical to that obtained for acid-extracted acrosin. Slab-gel electrophoresis under nondenaturing conditions showed only one active band, as revealed with a highly sensitive assay using N alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester as substrate. The radiation inactivation size of acid extracted acrosin was calculated to be 8400. This small unit could represent the active polypeptide portion of a larger monomer molecule or could represent the size of active subunits. Because acrosin is autocatalytic and highly active during fertilization, it is suggested that the active portion of the completely processed form of the enzyme is of small molecular weight.

Acids↗
PubMed1987

Outcome of mammary capsulotomies.

We reviewed the outcome of capsulotomies, both open and closed, in 134 patients with fibrous capsular contractures surrounding mammary implants. The rate of occurrence of fibrous capsules is given, along with other percentages for each type of augmentation. The study shows that the first attempt at a capsulotomy, either closed or open, gives a better result than the following attempts. After the third attempt, successful capsulotomies are either very rare or nonexistent. We therefore advise that after two or at most three unsuccessful closed capsulotomies, one should opt for an open capsulotomy. After two or at most three unsuccessful open capsulotomies, one should change to another approach.

Breast↗
PubMed1987

Characterization of D2 dopamine receptors in dopamine-resistant prolactin-secreting rat pituitary tumors 7315a and MtTW15.

We have investigated the structure of D2 receptors present in two prolactin-secreting, dopamine-resistant, transplantable rat pituitary tumors, 7315a and MtTW15. These receptors specifically bind with high affinity the dopamine antagonist [3H]spiroperidol when membrane bound or solubilized by [3-(3-cholamidopropyl)-dimethyl-ammonio]-1-propane sulfonate 10 mM and are pharmacologically characterized as D2 type. Target-size analysis by radiation inactivation indicated a molecular mass of approximately 100,000 and 200,000 daltons for receptors present respectively in 7315a and MtTW15 tumors either membrane bound or solubilized. The minimal size of the D2 binding site was evaluated at 94,000 daltons by photoaffinity labeling with [125I]azido-N-(p-aminophenethyl)-spiperone followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A guanine nucleotide had no effect on the displacing potency of the agonist N-propylnorapomorphine evaluated with membrane-bound or solubilized receptors obtained from either tumor. These results suggest the absence or inactivation of a guanine nucleotide binding protein in the receptorial complex of these tumors. Thus, our data indicate that a structural anomaly is present in the D2 receptorial complex of these prolactin-secreting rat pituitary tumors, which may be responsible for their resistance to the inhibitory effects of dopamine.

Affinity Labels↗
PubMed1986

Solubilization and characterization of D2-dopamine receptors in an estrone-induced, prolactin-secreting rat pituitary adenoma.

D2-dopamine (3,4-dihydroxyphenylethylamine) receptors were successfully solubilized with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate from an estrone-induced rat pituitary adenoma. Forty-five percent of initial protein and 48% of initial [3H]spiroperidol binding sites were solubilized. The high affinity as well as the stereoselectivity of the sites was preserved. The order of potency of dopaminergic agonists was found to be typical of D2 receptors. Target size analysis by radiation inactivation indicated a molecular weight of 143,000 +/- 3,000 and of 106,000 +/- 4,000 daltons for membrane-bound and solubilized receptors, respectively. This suggests the loss of a 37,000-dalton subunit during solubilization without significant modification of binding characteristics. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of receptor protein preparation photolabeled with N-(p-azido-m[125I]iodophenethyl)spiroperidol confirmed the existence of a 94,000-dalton peptide which probably constitutes the ligand binding site of the receptor. Thus, our data indicate that chronic estrogen treatment of rats, although inducing a pituitary adenoma, does not modify the pharmacological characteristics of D2 receptors. These data suggest therefore that these adenoma may represent an ideal source of material for further biochemical characterization of D2 receptors.

Adenoma↗
PubMed1985

Reduction mammoplasty by the total dermoglandular pedicle.

The use of a total dermoglandular pedicle in 700 cases of reduction mammoplasty are described by this team of authors. They emphasize that their modification of the dermoglandular pedicle provides an extremely reliable blood supply to the areola and is quick and easy to perform. There is no anesthesia of the nipple postoperatively, and the shape of the breast is favorable both in the immediate postoperative period and over the long-term follow-up observation between the years 1979 and 1984.

Adolescent↗
PubMed1985

Temperature dependence of the radiation inactivation of proteins.

The radiation inactivation method allows determination of the relative molecular mass (Mr) of proteins by exposure to high doses of ionizing radiation. The analysis by target theory of biological activity decay curves yields the size of the protein. A correction factor for Mr has been routinely used in the literature when irradiation is conducted at low temperature. Since the radiation inactivation of proteins is affected by temperature, we propose a general equation which relates Mr of a protein to D37,t, the dose in megarads at a given temperature t (in degree C) where 37% of its initial biological activity remains log Mr = 5.89 - log D37,t - 0.0028t. It is concluded that temperature affects the amount of absorbed radiation energy required to inactivate 1 mol of protein.

Mathematics↗
PubMed1984

Radiation inactivation of membrane proteins: molecular weight estimates in situ and after Triton X-100 solubilization.

Target size analysis by radiation inactivation is widely used for molecular weight determination of membrane enzymes and receptors in situ without the need for prior solubilization or purification. However, since most molecular weight data available in the literature on membrane proteins involve the use of detergents for solubilization, the target sizes of membrane proteins in situ and after solubilization by detergent treatment have been compared. Using data from the literature and personal results, three different types of behavior of membrane proteins in presence of detergents were found: (i) uncoupling of subunits (electric eel acetylcholinesterase, placental steroid sulfatase, and human nonspecific beta-glucosidase); (ii) coupling of protein molecules (mouse liver neuraminidase, and rat liver insulin receptor regulatory component); and (iii) no major change in quaternary structure (rat liver insulin receptor, kidney gamma-glutamyltransferase, asialoglycoprotein receptor, insulin degrading enzyme, and human leucocyte neuraminidase). For all these proteins, there is a statistically significant increase in target size of about 24% over the value obtained in situ without detergent. A relatively large body of literature data involving a variety of membrane proteins, membrane types, and irradiation conditions (electron accelerators or 60Co sources, and proteins irradiated in lyophilized form or frozen solution) was examined, and it was concluded that target sizes of membrane proteins, irradiated in the presence of Triton X-100, should be diminished by a factor of about 24% to obtain the molecular weight value.

Animals↗
PubMed1983

The molecular weight of the calcium-transport-ATPase of the human red blood cell determined by radiation inactivation.

Radiation inactivation was applied to analyze the molecular weight of the functional unit of (Ca2+ + Mg2+)-ATPase in human erythrocyte membranes. The enzyme activity was stable for at least 7 days at room temperature in membranes lyophilized in the presence of sucrose (150-300 mM). The enzyme activity in the lyophilized membranes and remaining after irradiation from a 60Co source was activated by calmodulin. A Mr of 290,000 +/- 15,000 was determined for (Ca2+ + Mg2+)-ATPase activity. Since the Mr by SDS-polyacrylamide gel electrophoresis is approximately 138,000 (Niggli et al. (1979) J. Biol. Chem. 254, 9955-9958), our results suggest that the Ca2+ pump ATPase functions as a dimer in the native human erythrocyte membrane.

Ca(2+) Mg(2+)-ATPase↗
PubMed1983

Target size analysis by radiation inactivation: a large capacity tube rack for irradiation in a Gammacell 220.

Target size analysis by radiation inactivation is now a well-established method to study structure-function relationships in biologically active macromolecules without prior purification or even solubilization. Recently, it was reported that a relatively low-dose-rate but commonly available gamma source such as the Gammacell 220 (Atomic Energy of Canada, Ltd.) can be used to carry out radiation inactivation experiments providing it is appropriately calibrated with enzymes of known radiation sensitivities (G. Beauregard and M. Potier (1982) Anal. Biochem. 122, 379-384). In this report, a tube rack designed to fit into the irradiation chamber of the Gammacell 220 which allows five experiments (at 30 tubes per experiment) to be carried out simultaneously with both standard and unknown samples is described. The dose rates delivered at different positions in the rack were determined by irradiating rat liver cytosolic neuraminidase, an enzyme of known radiation sensitivity. A better than 2.7% agreement was obtained between experimental dose rate and computed values from isodose curves previously published by other authors (O. A. Curzio and H. O. Quaranta (1982) Int. J. Appl. Radiat. Isot. 33, 1-3).

Animals↗
PubMed1983

The target sizes of the in situ and solubilized forms of human placental steroid sulfatase as measured by radiation inactivation.

Whole microsomal membrane preparations and Triton X-100-solubilized human placental steroid sulfatase were subjected to radiation inactivation analysis using gamma rays from a 60Co irradiator in order to assess the size of the physiological form of the enzyme. The data indicate that the enzyme exists as a monomer of molecular weight 78600 in Triton-containing buffers and as a polymer of molecular weight 533000 within the microsomal membrane.

Female↗
PubMed1982

Characterization of human placental neuraminidases. Stability, substrate specificity and molecular weight.

1. At least two components of neuraminidase can be distinguished on the basis of thermolability and sedimentability by using the artificial fluorogenic substrate 4-methylumbelliferyl N-acetyl-alpha-D-neuraminate. 2. In crude homogenates, thermodenaturation at 25 degrees C showed a biphasic curve corresponding to component A (half-life, 21 min) and B (half-life, 85 min). The two components were partially resolved by centrifugation. A being soluble and B sedimentable. Both had similar pH-activity curves (pH optimum, 4.4), Km values (A, 0.10 mM; B, 0.06 mM) and molecular weight as determined by radiation inactivation (A, 67000; B, 63000). 3. The soluble A form was still aggregated or bound to membranous debris since almost all neuraminidase activity was eluted near or at the void volume of a Sephacryl S-300 column. 4. Both soluble and sedimentable fractions of placenta hydrolysed the GD1A ganglioside and N-acetyl-neuraminyl-D-lactose linearly for 12 h but no fetuin hydrolysis was detected. 5. The neuraminidase activity with the artificial fluorogenic substrate was inhibited by N-acetylneuraminyl-D-lactose but not by the GD1A ganglioside. These preliminary results suggest that there exist two closely related enzymes hydrolysing both the artificial substrate and N-acetylneuraminyl-D-lactose and a third one hydrolysing the GD1A ganglioside exclusively.

Chromatography, Gel↗
PubMed1981

Familial articular chondrocalcinosis in Quebec.

The existence of articular chondrocalcinosis was documented in 9 members of 3 generations of a Quebec family. No associated or secondary forms of the disease were found. The clinical manifestations appeared early in life, and extensive radiologic involvement was apparent. We determined that genetic transmission was dominant, either autosomal or sex-linked, and not related to the HLA system.

Chondrocalcinosis↗
PubMed1980

Studies on the sialidoses: properties of human leucocyte neuraminidases.

At least two components of neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18) can be distinguished in human leucocytes on the basis of pH optimum, thermolability at 30 degrees C and the effect of the detergent octyl-beta-D-glucoside. With 4-methylumbelliferyl-alpha-D-N-acetylneuraminate as substrate, the A component has a pH optimum of 5.0, is labile at 30 degrees C and is unaffected by 0.2 M octyl-beta-glucoside. The B component has a pH optimum of 4.0-4.2, is stable at 30 degrees C but loses most of its activity in the presence of 0.2 M octyl-beta-glucoside. Both A and B components are membrane-bound but only the A component is solubilized by octyl-beta-glucoside in an active form. Molecular weights of neuraminidases by gamma-ray radiation inactivation (a method that does not require solubilization of the enzyme) were found to be 240 000 +/- 19 000 for the B component, 203 000 +/- 17 000 for the A component and 238 000 +/- 8000 for the octyl-beta-glucoside-solubilized A component. Gel filtration of soluble A component on Sephacryl S-300, in the presence of octyl-beta-glucoside, showed a single peak of activity eluted at or near the void volume suggesting that the enzyme is still in an aggregated form. Profound deficiency of neuraminidase activity was found for both A and B components in leucocytes of patients affected with sialidoses type 1 and 2 (less than 15% normal) and intermediate activity in obligate heterozygotes. These results suggest that the A and B components of leucocyte neuraminidase are closely related from the genetic point of view and that rapid diagnosis of sialidoses can be done by fluorimetric assay of neuraminidase in leucocytes.

Adolescent↗