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Studies on the sarcoplasmic reticulum (Ca²+ + Mg²+) ATPase / Bousselsela, Haoues 

Public ISBD Titre : Studies on the sarcoplasmic reticulum (Ca²+ + Mg²+) ATPase Type de document : texte imprimé Auteurs : Bousselsela, Haoues, Auteur ; Thomas, E. W., Directeur de thèse Editeur : University of Salford Année de publication : 1992 Importance : 216 f. Présentation : ill. Format : 27 cm. Note générale : Thèse d’État : Biologie : Salford, University of Salford : 1992 Bibliogr. f. 217 - 231 Langues : Anglais (eng) Mots-clés : ATPase  enzyme  ;  Inhibition  labelling  ;  Carbodiimides  ;  Phenolic  compounds  ;  Cyclopiazonic Index. décimale : D000992 Résumé : Sarcoplasmic reticulum vesicles were isolated from rabbit hindleg and back muscle, yielding 120-130 mg SR protein using 200-300 g muscle, and resulting in a(Ca²+ + Mg²+)- ATPase with specific activity of 13-15 µmol/mg/min. NCD-4 and NCD-5 were shown to be efficient as fluorescent probes of (Ca²+ + Mg²+)-ATPase. Both carbodiimides inhibited the enzyme activity only in the absence of Ca²+ (specificially labelled vesicles): inclusion of Ca²+(1 mM) protects against inhibition. A series of phenolic compounds showed an inhibitory action on (Ca²+ + Mg²+)-ATPase activity in the following order: bis (2-hydroxy-3, 5-methylphenyl) methane> 3(4'-hydroxyphenyl) decane and (4'-hydroxyphenyl-n-heptylketone> 1,1 bis(4-hydroxyphenyl) cyclohexane> 4,4'-isopropylidene diphenol> bis (2-hydroxyphenyl)methane> 4,4'-hydroxybenzophenone> 4-methoxyphenyl-n-heptylketone. No good correlation was obtained between the ability of phenols to inhibit (Ca²+ + Mg²+)-ATPase activity and the ability to perturb a Ca²+ -induced fluorescence quench leading to the suggestion that phenolic compounds affect other steps in the catalytic cycle besides those associated with Ca²+ -binding. Cyclopiazonic acid was found to be an excellent inhibitor whereas its analogs "A" and "B" showed no effect on (Ca²+ + Mg²+)-ATPase activity: this may possibly ascribed to the high ATP concentration used in the essay. All flavanoids examined exhibited an inhibitory effect on the (Ca²+ + Mg²+)-ATPase activity in the following order: robinetin> quercetin> luteolin> fisetin>galangin> rhamentin. This group of compounds required for their potency a 7-hydroxyl group but in the presence of additional hydroxyl groups at 3'4' and 5' positions. Hydroxyl groups 3 and 5 seem do not participate in the inhibitory potency of the flavonoids. Quercetin was found to inhibit (Ca²+ + Mg²+)-ATPase activity reversibly with half-maximal inhibition at 10 µM. This inhibition showed a pH dependence with the doses of quercetin being significantly lower at pH 8 than at pHs 7 and 6.4. Also this inhibition can be removed by affinity chromatography on a Reactive Red-120 agarose matrix. Irradiation with light of wavelengths> 350 nm of a mixture of (Ca²+ + Mg²+)-ATPase and quercetin led to covalent inhibition of enzyme, whose extent is related to quercetin and enzyme concentrations. Pre-irradiation of quercetin followed by incubation at room temperature for 30 min with (Ca²+ + Mg²+)-ATPase did not inhibit enzyme. The possibility of photolabelling of the enzyme is dicussed. Studies on the sarcoplasmic reticulum (Ca²+ + Mg²+) ATPase [texte imprimé] / Bousselsela, Haoues, Auteur ; Thomas, E. W., Directeur de thèse . - [S.l.] : University of Salford, 1992 . - 216 f. : ill. ; 27 cm. Thèse d’État : Biologie : Salford, University of Salford : 1992 Bibliogr. f. 217 - 231 Langues : Anglais (eng) Mots-clés : ATPase  enzyme  ;  Inhibition  labelling  ;  Carbodiimides  ;  Phenolic  compounds  ;  Cyclopiazonic Index. décimale : D000992 Résumé : Sarcoplasmic reticulum vesicles were isolated from rabbit hindleg and back muscle, yielding 120-130 mg SR protein using 200-300 g muscle, and resulting in a(Ca²+ + Mg²+)- ATPase with specific activity of 13-15 µmol/mg/min. NCD-4 and NCD-5 were shown to be efficient as fluorescent probes of (Ca²+ + Mg²+)-ATPase. Both carbodiimides inhibited the enzyme activity only in the absence of Ca²+ (specificially labelled vesicles): inclusion of Ca²+(1 mM) protects against inhibition. A series of phenolic compounds showed an inhibitory action on (Ca²+ + Mg²+)-ATPase activity in the following order: bis (2-hydroxy-3, 5-methylphenyl) methane> 3(4'-hydroxyphenyl) decane and (4'-hydroxyphenyl-n-heptylketone> 1,1 bis(4-hydroxyphenyl) cyclohexane> 4,4'-isopropylidene diphenol> bis (2-hydroxyphenyl)methane> 4,4'-hydroxybenzophenone> 4-methoxyphenyl-n-heptylketone. No good correlation was obtained between the ability of phenols to inhibit (Ca²+ + Mg²+)-ATPase activity and the ability to perturb a Ca²+ -induced fluorescence quench leading to the suggestion that phenolic compounds affect other steps in the catalytic cycle besides those associated with Ca²+ -binding. Cyclopiazonic acid was found to be an excellent inhibitor whereas its analogs "A" and "B" showed no effect on (Ca²+ + Mg²+)-ATPase activity: this may possibly ascribed to the high ATP concentration used in the essay. All flavanoids examined exhibited an inhibitory effect on the (Ca²+ + Mg²+)-ATPase activity in the following order: robinetin> quercetin> luteolin> fisetin>galangin> rhamentin. This group of compounds required for their potency a 7-hydroxyl group but in the presence of additional hydroxyl groups at 3'4' and 5' positions. Hydroxyl groups 3 and 5 seem do not participate in the inhibitory potency of the flavonoids. Quercetin was found to inhibit (Ca²+ + Mg²+)-ATPase activity reversibly with half-maximal inhibition at 10 µM. This inhibition showed a pH dependence with the doses of quercetin being significantly lower at pH 8 than at pHs 7 and 6.4. Also this inhibition can be removed by affinity chromatography on a Reactive Red-120 agarose matrix. Irradiation with light of wavelengths> 350 nm of a mixture of (Ca²+ + Mg²+)-ATPase and quercetin led to covalent inhibition of enzyme, whose extent is related to quercetin and enzyme concentrations. Pre-irradiation of quercetin followed by incubation at room temperature for 30 min with (Ca²+ + Mg²+)-ATPase did not inhibit enzyme. The possibility of photolabelling of the enzyme is dicussed.

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Comparative morphological, biochemical and immunological studies on two strains of vibrio anguillarum pathogenic in Salmo trutta / Said Dermime 

Public ISBD Titre : Comparative morphological, biochemical and immunological studies on two strains of vibrio anguillarum pathogenic in Salmo trutta Type de document : texte imprimé Auteurs : Said Dermime, Auteur ; J. B. Alexander, Auteur Editeur : University of Salford Année de publication : 1992 Importance : 269 f. Présentation : ill. Format : 30 cm. Note générale : Thèse de Doctorat: Biologie : United Kingdom, University of Salford : 1992 Bibliogr. f. 270 - 347 Langues : Anglais (eng) Mots-clés : Comparative  morphological Biochemical Immunological  studies  on  two  strains Vibrio  anguillarum  pathogenic Salmo  trutta Fish  immunopathology Index. décimale : D001192 Résumé : Two strains of vibrio anguillarum NCMB571 serotype 01 and NCMB829 serotype 02 were studied. Strain NCMB829 grew more vigorously than NCMB571. Bth weregram-negative rods with a single flagellum. Most NCMB829 cells were thin straight rods whilst NCMB571 were thick and curved. They were obligate, slight halophiles; true lactose fermenters and very sensitive to O/129. They both metabolised glucose and sucrose and NCMB571 also fermented arabinose. Arginine but not ornithine or lysine was metabolised by both strains. The serotypic differences of the two strains were confirmed and were due to the O-specific polysaccharide. A capsule was present in NCMB571 but the results for NCMB829 were equivocal. The antigenic components were constituents of the cell envelopes. When these were separated, the hetero-immunogenicity between the strains was found to be due to O-specific polysaccharides with different molecular weights. Cross-reactivity between the strains was due to common antigenic determinants present in a 47kDa polypeptide in NCMB829 and 51.9kDa in NCMB571. The primary humoral immune response of Salmo trutta to formalin-killed and heat-killed NCMB829 AND NCMB571 was measured by indirect ELISA. With both strains, formalin-killed bacteria stimulated more antibody production than heat-killed ones. The efficacy of the bacterial antigens and the possibility of cross-protection and antigenic competition were studied in vaccinated brown trout. The homologous monovalent and the bivalent bacterins were highly effective in protecting fish from vibriosis. However, the cross-protection between the groups of fish that received heterologous monovalent bacterin was low. Antigenic competition between the strains was not observed. A secondary humoral immune reponse in brown trout after vaccination with bivalent bacterin and challenged with live bacteria was demonstrated and some correlation between antibody production and protection was observed Comparative morphological, biochemical and immunological studies on two strains of vibrio anguillarum pathogenic in Salmo trutta [texte imprimé] / Said Dermime, Auteur ; J. B. Alexander, Auteur . - [S.l.] : University of Salford, 1992 . - 269 f. : ill. ; 30 cm. Thèse de Doctorat: Biologie : United Kingdom, University of Salford : 1992 Bibliogr. f. 270 - 347 Langues : Anglais (eng) Mots-clés : Comparative  morphological Biochemical Immunological  studies  on  two  strains Vibrio  anguillarum  pathogenic Salmo  trutta Fish  immunopathology Index. décimale : D001192 Résumé : Two strains of vibrio anguillarum NCMB571 serotype 01 and NCMB829 serotype 02 were studied. Strain NCMB829 grew more vigorously than NCMB571. Bth weregram-negative rods with a single flagellum. Most NCMB829 cells were thin straight rods whilst NCMB571 were thick and curved. They were obligate, slight halophiles; true lactose fermenters and very sensitive to O/129. They both metabolised glucose and sucrose and NCMB571 also fermented arabinose. Arginine but not ornithine or lysine was metabolised by both strains. The serotypic differences of the two strains were confirmed and were due to the O-specific polysaccharide. A capsule was present in NCMB571 but the results for NCMB829 were equivocal. The antigenic components were constituents of the cell envelopes. When these were separated, the hetero-immunogenicity between the strains was found to be due to O-specific polysaccharides with different molecular weights. Cross-reactivity between the strains was due to common antigenic determinants present in a 47kDa polypeptide in NCMB829 and 51.9kDa in NCMB571. The primary humoral immune response of Salmo trutta to formalin-killed and heat-killed NCMB829 AND NCMB571 was measured by indirect ELISA. With both strains, formalin-killed bacteria stimulated more antibody production than heat-killed ones. The efficacy of the bacterial antigens and the possibility of cross-protection and antigenic competition were studied in vaccinated brown trout. The homologous monovalent and the bivalent bacterins were highly effective in protecting fish from vibriosis. However, the cross-protection between the groups of fish that received heterologous monovalent bacterin was low. Antigenic competition between the strains was not observed. A secondary humoral immune reponse in brown trout after vaccination with bivalent bacterin and challenged with live bacteria was demonstrated and some correlation between antibody production and protection was observed

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Synthesis and photochemistry of α-substituted carbonyl compounds / Ahmed M'hamedi 

Public ISBD Titre : Synthesis and photochemistry of α-substituted carbonyl compounds Type de document : texte imprimé Auteurs : Ahmed M'hamedi, Auteur ; J. Hill, Directeur de thèse Editeur : University of Salford Année de publication : 1992 Importance : 138 f. Format : 30 cm. Note générale : Thèse de Doctorat : Génie Chimique : Salford, University of Salford : 1992 Bibliogr. f. 139 - 153 Langues : Anglais (eng) Mots-clés : α-substituted  carbonyl  compounds Photochemistry α-amino-enones Coumarins Irradiation α-N-Methylaniliosuccinimides Cyclisation Index. décimale : D001292 Résumé : The work described in this thesis deals with various aspects of the photochemistry of α-substituted carbonyl compounds, and is made up of three parts. 1. α-amino-enones: As part of studies into the photochemistry of α-amino-carbonyl compounds, α-amino-enones 1a and 1b and coumarins 4a and 4b were synthesised and irradiated. Irradiation of 1a yielded the azetidinol 3a and enone isomers 2a, and 1b gave impure azetidinol 3b and enone isomers 2b. On the other hand, coumarin 4a and 4b gave complex photoproduct mixtures. 2. α-N-methylanilinosuccinimides and related compounds 2-(N-mthylanilino)-N-substituted succinimides 5a-c were synthesised and irradiated. These compounds were prepared from maleic anhydride via the corresponding N-substituted maleimides. Irradiation of 5a-c gave the pararearrangement products 6a-c respectively, generally as the major photoproduct. The ortho-rearrangement product 7 was also obtained from 5a. No significant photoproducts were formed from N-benzylsuccinimide 8. 2-(N-Methylanilino)-N-phenylmaleimide 9, prepared from N-phenylmaleimide, was irradiated, but no product could be isolated from the complex photolysis mixture. The succinimide 11, prepared from itaconic anhydride 10, gave a fused-ring photoproduct 12 as a result of a type II cyclisation. 3. Unsaturated α-substituted carbonyl compounds: Radical cyclisations. A variety of α-substituted compounds were irradiated in a study into the use of such photoreactions as a method for radical cyclisations. a/ Unsaturated β-ketosulphides 13 and 14 were prepared. Irradiation gave the cyclisation product 17 and 18 respectively. Unsaturated β-ketoester-sulphides 15 and 16a-e were synthesised. Irradiation of 15 and 16d yielded the cyclisation products 19 and 20 respectively. However, irradiation of 16 a and 16 d gave complex mixtures from which no product could be isolated. b/ α-Substituted esters 22a-c were synthesised via chloroester 21. Irradiation of 22a and 22b yielded the non-cyclised products 23 and 24 respectively. No significant product aws produced on irradiation of 22c. c/ Aseries of α-substituted amides 25a-f were synthesised. Irradiation of 25a and 25d yielded the cyclisation products 26 and 27 respectively, whereas 25a gave diallylacetamide 28 on irradiation in the presence of benzyl mercaptan. Amides 25b, c, e and f gave the corresponding non-cyclised acetamides 29 a or b. Synthesis and photochemistry of α-substituted carbonyl compounds [texte imprimé] / Ahmed M'hamedi, Auteur ; J. Hill, Directeur de thèse . - [S.l.] : University of Salford, 1992 . - 138 f. ; 30 cm. Thèse de Doctorat : Génie Chimique : Salford, University of Salford : 1992 Bibliogr. f. 139 - 153 Langues : Anglais (eng) Mots-clés : α-substituted  carbonyl  compounds Photochemistry α-amino-enones Coumarins Irradiation α-N-Methylaniliosuccinimides Cyclisation Index. décimale : D001292 Résumé : The work described in this thesis deals with various aspects of the photochemistry of α-substituted carbonyl compounds, and is made up of three parts. 1. α-amino-enones: As part of studies into the photochemistry of α-amino-carbonyl compounds, α-amino-enones 1a and 1b and coumarins 4a and 4b were synthesised and irradiated. Irradiation of 1a yielded the azetidinol 3a and enone isomers 2a, and 1b gave impure azetidinol 3b and enone isomers 2b. On the other hand, coumarin 4a and 4b gave complex photoproduct mixtures. 2. α-N-methylanilinosuccinimides and related compounds 2-(N-mthylanilino)-N-substituted succinimides 5a-c were synthesised and irradiated. These compounds were prepared from maleic anhydride via the corresponding N-substituted maleimides. Irradiation of 5a-c gave the pararearrangement products 6a-c respectively, generally as the major photoproduct. The ortho-rearrangement product 7 was also obtained from 5a. No significant photoproducts were formed from N-benzylsuccinimide 8. 2-(N-Methylanilino)-N-phenylmaleimide 9, prepared from N-phenylmaleimide, was irradiated, but no product could be isolated from the complex photolysis mixture. The succinimide 11, prepared from itaconic anhydride 10, gave a fused-ring photoproduct 12 as a result of a type II cyclisation. 3. Unsaturated α-substituted carbonyl compounds: Radical cyclisations. A variety of α-substituted compounds were irradiated in a study into the use of such photoreactions as a method for radical cyclisations. a/ Unsaturated β-ketosulphides 13 and 14 were prepared. Irradiation gave the cyclisation product 17 and 18 respectively. Unsaturated β-ketoester-sulphides 15 and 16a-e were synthesised. Irradiation of 15 and 16d yielded the cyclisation products 19 and 20 respectively. However, irradiation of 16 a and 16 d gave complex mixtures from which no product could be isolated. b/ α-Substituted esters 22a-c were synthesised via chloroester 21. Irradiation of 22a and 22b yielded the non-cyclised products 23 and 24 respectively. No significant product aws produced on irradiation of 22c. c/ Aseries of α-substituted amides 25a-f were synthesised. Irradiation of 25a and 25d yielded the cyclisation products 26 and 27 respectively, whereas 25a gave diallylacetamide 28 on irradiation in the presence of benzyl mercaptan. Amides 25b, c, e and f gave the corresponding non-cyclised acetamides 29 a or b.

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