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Characterization of the sialic acid component in a bioactive extract from the edible bird's nest.

by Ng Ping-chung. / Thesis (M. Phil.)--Chinese University of Hong Kong, 1991. / Includes bibliographical references. / Chapter 1. --- Introduction / Chapter 1.1 --- Natural History of the Bird and the Nest --- p.1 / Chapter 1.2 --- What is More Important in Saliva: Mucin or Proteoglycan? --- p.9 / Chapter 1.3 --- Extraction and characterization of salivary glycoprotein --- p.27 / Chapter 2. --- Materials and Methods / Chapter 2.1 --- Preparation of Swiftlet's Nest Extracts --- p.38 / Chapter 2.2 --- Chemical and Biochemical Analysis of SN extracts --- p.38 / Chapter 2.2.1 --- Chemical Analysis --- p.38 / Chapter 2.2.1.1 --- Element Analysis --- p.38 / Chapter 2.2.1.2 --- Ash and Atomic Absorption Analysis --- p.39 / Chapter 2.2.2 --- Biochemical Analysis --- p.39 / Chapter 2.2.2.1 --- Protein Determination --- p.39 / Chapter 2.2.2.2 --- Hexose Determination --- p.40 / Chapter 2.2.2.3 --- Uronic Acid Determination --- p.40 / Chapter 2.2.2.4 --- Hexosamine Determination --- p.41 / Chapter 2.2.2.5 --- Sialic Acid Determination --- p.42 / Chapter 2.2.2.6 --- Sulphate Determination --- p.42 / Chapter 2.3 --- Assay of Co-mitogenic Activity in Lymphocyte Culture --- p.43 / Chapter 2.3.1 --- Preparation of Human Peripheral Blood Lymphocytes --- p.43 / Chapter 2.3.2 --- Co-mitogenic Assay --- p.43 / Chapter 2.4 --- Effect of SN pretreatment on Concanavalin A-stimulated Blastogenic Response in Mouse Splenocytes --- p.44 / Chapter 2.4.1 --- Administration of SN extracts --- p.44 / Chapter 2.4.2 --- Preparation of Mouse Splenocytes --- p.45 / Chapter 2.4.3 --- Concanavalin A-stimulated Blastogenic Response Assay --- p.45 / Chapter 2.5 --- Characterization of SN extracts by Chromatographic Methods --- p.45 / Chapter 2.5.1 --- Gel Filtration Chromatography --- p.45 / Chapter 2.5.1.1 --- Sephadex G-200 Chromatography --- p.45 / Chapter 2.5.1.2 --- Superose-Fast Protein Liquid Chromatography --- p.46 / Chapter 2.5.2 --- Ion-exchange Chromatography --- p.46 / Chapter 2.5.2.1 --- DEAE-Sepharose --- p.46 / Chapter 2.5.2.2 --- Mono-Q FPLC --- p.46 / Chapter 2.5.3 --- Immobilized Metal Affinity Chromatography --- p.47 / Chapter 2.5.4 --- Wheat-germ lectin Sepharose Chromatography --- p.47 / Chapter 2.5.5 --- Octyl-Sepharose Chromatography --- p.47 / Chapter 2.5.6 --- Limulus Polyemus Agarose Chromatography --- p.48 / Chapter 2.5.7 --- Heparin-Agarose Chromatography --- p.48 / Chapter 2.6 --- Electrophoretic Analysis of SN extract --- p.49 / Chapter 2.6.1 --- Sodium Dodecyl Sulphate/Polyacrylamide Gel Electrophoresis --- p.49 / Chapter 2.6.2 --- Isoelectrofocusing in Polyacrylamide Gel --- p.50 / Chapter 2.6.2.1 --- Preparation of Gel --- p.50 / Chapter 2.6.2.2 --- Isoelectrofocusing Procedure --- p.51 / Chapter 2.6.2.3 --- "Fixing, Staining and Destaining" --- p.51 / Chapter 2.6.3 --- Gradient Polyacrylamide Gel Electrophoresis --- p.51 / Chapter 2.6.3.1 --- Preparation of Gradient Polyacrylamide Gel --- p.52 / Chapter 2.7 --- Enzymatic Modification of SN extracts --- p.53 / Chapter 2.7.1 --- B-glucuronidase (EC 3.2.1.31) --- p.53 / Chapter 2.7.2 --- Hyaluronidase (EC 3.2.1.35) --- p.53 / Chapter 2.7.3 --- Chondroitinase ABC (EC 4.2.2.4) --- p.54 / Chapter 2.8 --- Miscellaneous Reagents --- p.54 / Chapter 2.8.1 --- Phosphate-buffer-saline (PBS) --- p.54 / Chapter 2.8.2 --- Fetal Calf Serum (FGS) --- p.54 / Chapter 2.8.3 --- Mitogen --- p.55 / Chapter 2.3.4 --- Penicillin-Streptomycin-Fungizone Solution --- p.55 / Chapter 2.8.5 --- RPMI-1640 Medium --- p.55 / Chapter 2.8.6 --- Scintillant --- p.55 / Chapter 2.8.7 --- Trypan Blue Solution --- p.55 / Chapter 3 --- Results --- p.57 / Chapter 3.1 --- Extraction of Biologically active fractions from swiflet's nest --- p.57 / Chapter 3.2 --- Effect of SN pretreatment on Con A-stimulated Blastogenesis response in mouse splenocytes --- p.57 / Chapter 3.3 --- Chemical and Biochemical Analysis of SN extracts --- p.57 / Chapter 3.3.1 --- Atomic absorption analysis of metal elements --- p.57 / Chapter 3.3.2 --- Element analysis --- p.61 / Chapter 3.3.3 --- Biochemical analysis --- p.61 / Chapter 3.4 --- Chromatographic characterization of SN extracts --- p.67 / Chapter 3.4.1 --- Sephadex G-200 chromatography --- p.67 / Chapter 3.4.2 --- DEAE-Sepharose chromatography --- p.67 / Chapter 3.4.3 --- Immobilized Metal Affinity Chromatography --- p.73 / Chapter 3.4.4 --- Wheat-germ lectin Sepharose Chromatography --- p.73 / Chapter 3.4.5 --- Limulus Polyemus Agarose Chromatography --- p.73 / Chapter 3.4.6 --- Octyl-Sepharose Chromatography --- p.81 / Chapter 3.4.7 --- Heparin-Agarose Chromatography --- p.81 / Chapter 3.5 --- Electrophoretic Analysis of SN extract --- p.88 / Chapter 3.5.1 --- Sodium Dodecyl Sulphate/Polyacrylamide Gel Electrophoresis --- p.88 / Chapter 3.5.2 --- Isoelectrofocusing in Thin Layer of Polyacrylamide Gel --- p.88 / Chapter 3.5.3 --- Gradient Polyacrylamide Gel Electrophoresis --- p.88 / Chapter 3.6 --- Enzymatic Modification of SN extracts --- p.92 / Chapter 3.6.1 --- B-glucuronidase (EC 3.2.1.31) --- p.92 / Chapter 3.6.1.1 --- Alteration in co-mitogenic activity --- p.92 / Chapter 3.6.1.2 --- Alternation in uronic acid content --- p.92 / Chapter 3.7 --- Co-mitogenic activity of glycosaminoglycans --- p.92 / Chapter 3.8 --- Effect of heparin on the co-mitogenic activity of SNp2C fraction --- p.96 / Chapter 4 --- Discussion --- p.99 / Chapter 4.1 --- Extraction of Biologically active fraction from edible bird's nest --- p.99 / Chapter 4.2 --- Chemical and Biochemical characterization of active component(s) in SN extract --- p.100 / Chapter 4.3 --- Some mechanistic considerations --- p.107 / Chapter 4.4 --- Summary and Conclusion --- p.111 / References --- p.114

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_318738
Date January 1991
ContributorsNg, Ping-chung., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
PublisherChinese University of Hong Kong
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, abstract or summary
Formatprint, [viii], 121 leaves : ill. (some col.) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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