Thesis (M.S.)--West Virginia University, 2001. / Title from document title page. Document formatted into pages; contains vi, 43 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 37-43).

Thesis (M.S.)--West Virginia University, 2005. / Title from document title page. Document formatted into pages; contains vii, 44 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 41-44).

No description available.

Thesis (M.S.)--West Virginia University, 2002. / Title from document title page. Document formatted into pages; contains vi, 50 p. : ill. Vita. Includes abstract. Includes bibliographical references (p. 43-48).

Zugl.: Halle, Wittenberg, University, Diss., 2001.

published_or_final_version / Botany / Master / Master of Philosophy

No description available.

One of the many recorded adaptive responses in respiratory chain complex I deficient cells is the over-expression of the small metal binding proteins, metallothioneins (MTs). The antioxidant properties of MTs putatively protect the deficient cells against oxidative damage, thus limiting further damage and impairment of enzymes involved in energy production. Moreover, the role of metallothioneins in supplying metal cofactors to enzymes and transcription factors in order to promote energy metabolism was previously proposed, which could accompany their role as antioxidants. This view is supported by the observations that MT knockout mice tend to become moderately obese, implying a lower energy metabolic rate. Hence, the involvement of metallothioneins in mitochondrial function and disease cannot be ignored. However, this association is still very vague due to the diversity of their functions and the complexity of the mitochondrion. The use of systems biology technology and more specifically metabolomics technology was thus employed to clarify this association by investigating the metabolic differences between wild type and MT knockout mice in unchallenged conditions as well as when mitochondrial function (energy metabolism) was challenged with exercise and/or a high-fat diet. The metabolic differences between these mice were also studied when complex I of the respiratory chain was inhibited with rotenone. The metabolome content of different tissues and bio-fluids were examined in an untargeted fashion using three standardized analytical platforms and the data mined using modern metabolomics and related statistical methods. Clear metabolic differences were found between the wild type and MT knockout mice during unchallenged conditions. These metabolic differences were persisted and were often amplified when mitochondrial metabolism was specifically challenged through exercise, high-fat intake or complex I inhibition. The data pointed to an overall reduced metabolic rate in the MT knockout mice and possible insulin resistance after the interventions which imply (and confirm) the involvement of MTs in promoting energy metabolism in the wild type mice. / Thesis (Ph.D. (Biochemistry))--North-West University, Potchefstroom Campus, 2012

Chan Pat Chun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 153-177). / Acknowledgments --- p.i / Presentations Derived from the Present thesis Work --- p.ii / Abstract --- p.iii / Abbreviations --- p.v / Abbreviation Table for Amino Acids --- p.viii / List of Figures --- p.ix / List of Tables --- p.xi / Contents --- p.xii / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Environmental Pollutants --- p.1 / Chapter 1.2 --- Pollutant Inducible Genes (PIGs) --- p.1 / Chapter 1.2.1 --- Classification of PIGS --- p.2 / Chapter 1.2.1.1 --- Drug Metabolizing Enzymes/Proteins --- p.2 / Chapter 1.2.1.2 --- Stress Proteins --- p.5 / Chapter 1.2.1.3 --- Antioxidant Enzymes --- p.6 / Chapter 1.2.1.4 --- "Hormones, Growth Factors and Their Receptors" --- p.6 / Chapter 1.2.1.5 --- Enzymes/Proteins Involved in Bioenergetics --- p.6 / Chapter 1.2.2 --- PIGs as a Field of Study --- p.8 / Chapter 1.2.2.1 --- Study of the Mechanism of Detoxification and Toxication --- p.8 / Chapter 1.2.2.2 --- Biomarker Study --- p.9 / Chapter 1.2.2.3 --- Study of Regulation of Gene Expression --- p.11 / Chapter 1.2.2.4 --- Study of Evolution --- p.12 / Chapter 1.3 --- Aims and Rational of the Present Study --- p.12 / Chapter 2 --- General Methodology --- p.15 / Chapter 2.1 --- Materials --- p.15 / Chapter 2.2.1 --- Reagents --- p.15 / Chapter 2.1.1.1 --- Preparation of Plasmid DNA --- p.15 / Chapter 2.1.1.2 --- Preparation of Genomic DNA --- p.15 / Chapter 2.1.1.3 --- Purification of Total RNA --- p.16 / Chapter 2.1.1.4 --- Restriction Enzyme Digestion --- p.16 / Chapter 2.1.1.5 --- Capillary Blotting of DNA (Southern Blotting) --- p.16 / Chapter 2.1.1.6 --- Capillary Blotting of Total RNA (Northern Blotting) --- p.17 / Chapter 2.1.1.7 --- Hybridization --- p.17 / Chapter 2.1.1.8 --- Library Screening --- p.18 / Chapter 2.1.1.9 --- Polymerase Chain Reaction --- p.18 / Chapter 2.1.1.10 --- Transformation of E. coli Competent Cells --- p.19 / Chapter 2.1.1.11 --- Nucleotide Sequence Determination --- p.19 / Chapter 2.1.2 --- List of Primers --- p.20 / Chapter 2.1.2.1 --- Primers used for Nucleotide Sequence Determination --- p.20 / Chapter 2.1.2.2 --- Primer Used for First Strand cDNA Synthesis --- p.20 / Chapter 2.1.2.3 --- Primers for Amplifying Actin cDNA Fragment --- p.20 / Chapter 2.1.2.4 --- Common Carp MT Specific Primers --- p.20 / Chapter 2.1.2.5 --- Teleost CYP1A Specific Primers --- p.21 / Chapter 2.1.2.6 --- Common Carp CYP1A Specific Primers --- p.21 / Chapter 2.1.2.7 --- Primers and Cassettes for the Cloning of5' Upstream Regions of MT Genes --- p.21 / Chapter 2.1.3 --- Accession Numbers of Selected P450 and MT Nucleotide and Amino Acid Sequences in the Genebank --- p.21 / Chapter 2.1.3.1 --- MTs of Different Teleost Species --- p.21 / Chapter 2.1.3.2 --- MTs of Other Vertebrate Species' --- p.22 / Chapter 2.1.3.3 --- P450s of Different Teleost Species --- p.22 / Chapter 2.1.3.4 --- CYP1s of Different Mammalian Species --- p.22 / Chapter 2.2 --- Methods --- p.23 / Chapter 2.2.1 --- Preparation of Plasmid --- p.23 / Chapter 2.2.2 --- Preparation of Genomic DNA --- p.23 / Chapter 2.2.3 --- Purification of Total RNA --- p.24 / Chapter 2.2.4 --- Restriction Enzyme Digestion --- p.25 / Chapter 2.2.5 --- Capillary Blotting of DNA (Southern Blotting) --- p.25 / Chapter 2.2.5.1 --- Semi-dry Capillary Blotting --- p.25 / Chapter 2.2.5.2 --- Alkaline Transfer --- p.25 / Chapter 2.2.5.3 --- Transfer of Digested Genomic DNA on to Nylon Membrane --- p.26 / Chapter 2.2.6 --- Capillary Blotting of Total RNA (Northern Blotting) --- p.26 / Chapter 2.2.7 --- Radioactive Labeling of Nucleic Acid Probes --- p.26 / Chapter 2.2.8 --- Hybridization --- p.27 / Chapter 2.2.9 --- Library Screening --- p.27 / Chapter 2.2.9.1 --- Construction of Liver cDNA Library of Adult Common Carp --- p.27 / Chapter 2.2.9.2 --- Preparation of Plating Cells --- p.27 / Chapter 2.2.9.3 --- Phage Tittering --- p.27 / Chapter 2.2.9.4 --- Primary Screening --- p.28 / Chapter 2.2.9.5 --- Secondary Screening / Chapter 2.2.9.6 --- Conversion of Phage DNA to Phagemid by invivo Excision --- p.28 / Chapter 2.2.10 --- First Strand cDNA Synthesis --- p.29 / Chapter 2.2.11 --- Polymerase Chain Reaction --- p.29 / Chapter 2.2.12 --- Ligation of DNA with Linearized Plasmid --- p.30 / Chapter 2.2.13 --- Transformation of E. coli Competent Cell --- p.30 / Chapter 2.2.14 --- Nucleotide Sequence Determination --- p.31 / Chapter 2.2.15 --- Densitometric Analysis --- p.31 / Chapter 3 --- "Cloning of Common Carp MT cDNA and Gene, and Induction of MT mRNA Expression" --- p.32 / Chapter 3.1 --- Introduction --- p.32 / Chapter 3.1.1 --- Metals in Biological System --- p.32 / Chapter 3.1.2 --- Metallothionein --- p.33 / Chapter 3.1.2.1 --- Functions of MT --- p.26 / Chapter 3.1.2.2 --- Regulation of MT Expression --- p.39 / Chapter 3.1.3 --- Fish MTs --- p.44 / Chapter 3.1.3.1 --- Detection of MT in Teleost --- p.46 / Chapter 3.1.3.2 --- MT Studies in Common Carp --- p.47 / Chapter 3.1.4 --- Specific Aims of This Chapter --- p.49 / Chapter 3.2 --- Strategies --- p.50 / Chapter 3.3 --- Specific Methods --- p.50 / Chapter 3.3.1 --- Cloning of MT cDNAs of Common Carp --- p.50 / Chapter 3.3.2 --- Analysis of MT cDNA Sequences --- p.51 / Chapter 3.3.3 --- Southern Blot Analysis of Common Carp Genomic DNA --- p.52 / Chapter 3.3.4 --- Amplification of MT Gene Fragments Using PCR --- p.52 / Chapter 3.3.5 --- Amplification of the 5' Upstream Regions of MT Genes Using PCR --- p.52 / Chapter 3.3.6 --- Endogenous MT mRNA Expression of Juvenile and Adult Common Carp --- p.54 / Chapter 3.3.7 --- Induction of MT mRNA of Juvenile Common Carp Injected with Cadmium --- p.55 / Chapter 3.4 --- Results --- p.56 / Chapter 3.4.1 --- Cloning of Common Carp MT cDNAs --- p.56 / Chapter 3.4.2 --- Analysis of the MT cDNA Sequences --- p.57 / Chapter 3.4.3 --- Southern Blot Analysis of the Common Carp Genomic DNA --- p.59 / Chapter 3.4.4 --- Amplification of the MT Gene Fragments of Common Carp Using PCR --- p.62 / Chapter 3.4.5 --- Amplification of the 5' Upstream Regions of MT Genes using PCR --- p.65 / Chapter 3.4.6 --- Endogenous MT mRNA Expression of Juvenile and Adult Common Carp --- p.67 / Chapter 3.4.7 --- Induction of MT mRNA of Juvenile Common Carp Injected with Cadmium --- p.68 / Chapter 3.5 --- Discussion --- p.72 / Chapter 3.5.1 --- MT cDNAs of Common Carp --- p.72 / Chapter 3.5.1.1 --- Coding Region --- p.72 / Chapter 3.5.1.2 --- The 3' Untranslated Region --- p.75 / Chapter 3.5.1.3 --- The 5' Untranslated Region --- p.76 / Chapter 3.5.2 --- MT Genes of Common Carp --- p.77 / Chapter 3.5.3 --- MT mRNA Expression of Common Carp --- p.82 / Chapter 3.5.4 --- Normalization of the Signals of Northern Blot Analysis --- p.85 / Chapter 3.5.5 --- Common Carp MT mRNA as Biomarker of Heavy Metal Exposure? --- p.87 / Chapter 3.6 --- Conclusion --- p.89 / Chapter 4 --- Cloning of Common Carp CYP1A cDNAs and Induction of CYP1A mRNA Expression --- p.90 / Chapter 4.1 --- Introduction --- p.90 / Chapter 4.1.1 --- Cytochrome P450s --- p.90 / Chapter 4.1.2 --- Cytochrome P450 1 (CYP1) --- p.93 / Chapter 4.1.3 --- AhR Mediated CYP1A1 Gene Induction --- p.94 / Chapter 4.1.3.1 --- Anthropogenic Sources of AhR Ligands --- p.95 / Chapter 4.1.3.2 --- Natural Sources of AhR Ligands --- p.97 / Chapter 4.1.3.3 --- Potency of Inducibility --- p.97 / Chapter 4.1.3.4 --- Induction of CYP1A1 Gene Transcription by AhR --- p.98 / Chapter 4.1.3.5 --- Non-AhR Mediated CYP1A1 Gene Transcription? --- p.105 / Chapter 4.1.4 --- CYP1A Studies in Teleost Species --- p.107 / Chapter 4.1.4.1 --- Regulation of CYP1A in Teleost --- p.109 / Chapter 4.1.4.2 --- Detection of CYP1A in Teleost --- p.111 / Chapter 4.1.4.3 --- CYP1A Studies of Common Carp --- p.113 / Chapter 4.1.5 --- Specific Aims of This Chapter --- p.114 / Chapter 4.2 --- Strategies --- p.115 / Chapter 4.3 --- Specific Methods --- p.119 / Chapter 4.3.1 --- RT-PCR of CYP1A cDNAs of Common Carp --- p.119 / Chapter 4.3.2 --- Determination of the Nucleotide Sequences of the CYP1A cDNAs of Common Carp --- p.119 / Chapter 4.3.3 --- Library Screening --- p.119 / Chapter 4.3.4 --- Analysis of the CYP1A Genes of Common Carp --- p.121 / Chapter 4.3.5 --- Induction of CYP1A mRNA of Common Carp Injected with 3-MC --- p.122 / Chapter 4.4 --- Results --- p.123 / Chapter 4.4.1 --- RT-PCR of CYP1A cDNAs of Common Carp --- p.123 / Chapter 4.4.2 --- Determination of the Nucleotide Sequences of the CYP1A cDNAs of Common Carp --- p.124 / Chapter 4.4.3 --- Library Screening --- p.124 / Chapter 4.4.4 --- Analysis of the CYP1A Genes of Common Carp --- p.128 / Chapter 4.4.5 --- Induction of CYP1A mRNA of Common Carp Injected with 3-MC --- p.131 / Chapter 4.5 --- Discussion --- p.134 / Chapter 4.5.1 --- On the Use of Rainbow Trout CYP1A1 cDNA Probe --- p.134 / Chapter 4.5.2 --- CYP1A cDNAs of Common Carp --- p.134 / Chapter 4.5.3 --- CYP1A Genes of Common Carp --- p.138 / Chapter 4.5.4 --- CYP1A Expression in Uninduced and Induced Tissues --- p.142 / Chapter 4.5.5 --- The Use of CYP1A cDNAs As Biomarkers --- p.146 / Chapter 4.6 --- Conclusion --- p.148 / Chapter 5 --- General Conclusion --- p.149 / Chapter 6 --- References --- p.153

One of the many recorded adaptive responses in respiratory chain complex I deficient cells is the over-expression of the small metal binding proteins, metallothioneins (MTs). The antioxidant properties of MTs putatively protect the deficient cells against oxidative damage, thus limiting further damage and impairment of enzymes involved in energy production. Moreover, the role of metallothioneins in supplying metal cofactors to enzymes and transcription factors in order to promote energy metabolism was previously proposed, which could accompany their role as antioxidants. This view is supported by the observations that MT knockout mice tend to become moderately obese, implying a lower energy metabolic rate. Hence, the involvement of metallothioneins in mitochondrial function and disease cannot be ignored. However, this association is still very vague due to the diversity of their functions and the complexity of the mitochondrion. The use of systems biology technology and more specifically metabolomics technology was thus employed to clarify this association by investigating the metabolic differences between wild type and MT knockout mice in unchallenged conditions as well as when mitochondrial function (energy metabolism) was challenged with exercise and/or a high-fat diet. The metabolic differences between these mice were also studied when complex I of the respiratory chain was inhibited with rotenone. The metabolome content of different tissues and bio-fluids were examined in an untargeted fashion using three standardized analytical platforms and the data mined using modern metabolomics and related statistical methods. Clear metabolic differences were found between the wild type and MT knockout mice during unchallenged conditions. These metabolic differences were persisted and were often amplified when mitochondrial metabolism was specifically challenged through exercise, high-fat intake or complex I inhibition. The data pointed to an overall reduced metabolic rate in the MT knockout mice and possible insulin resistance after the interventions which imply (and confirm) the involvement of MTs in promoting energy metabolism in the wild type mice. / Thesis (Ph.D. (Biochemistry))--North-West University, Potchefstroom Campus, 2012