Thesis (Ph. D.)--Cornell University, May, 2007. / Vita. Includes bibliographical references (leaves 273-295).

Dissertation (Ph.D.)--University of Toledo, 2008. / "In partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biomedical Sciences." Title from title page of PDF document. Bibliography: p. 126-146.

Thesis (Ph.D. in Pharmacology) -- University of Colorado Denver, 2007. / Typescript. Includes bibliographical references (leaves 105-113). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;

Diss. (sammanfattning) Stockholm : Stockholms universitet, 2008. / Härtill 4 uppsatser.

Thesis (Ph. D.)--University of Texas at Austin, 2007. / Vita. Includes bibliographical references.

Metal pollution has been a great challenge in most industrialized countries as a result of waste generated from industrial activities being introduced into the environment. Unicellular green algae have been considered a potential biological tool for bioremediation of metal pollutants due to its metal sequestration properties. However, methods for further improving unicellular green algae metal sequestration by manipulating metal uptake and tolerance in unicellular green algae have not been studied in detail. In this study, a family metal transport protein named MTP1 - MTP4 from C. reinhardtii were screened by yeast heterologous expression for metal transport activity. MTP1 was able to strongly rescue the Zn and Co sensitivity of the zrc1cot1 strain, MTP3 could weakly mediate Zn and Co growth, but MTP2 and MTP4 appeared to have no Zn or Co tolerance activity. MTP2, MTP3 and MTP4 but not MTP1 could strongly rescue the Mn sensitivity of the pmr1 strain. When MTP4 was over-expressed in C. reinhardtii the strain showed a significant increase in Cd tolerance compared to the wild type, but no significant difference in Mn tolerance and uptake. AtHMA4 a Zn2+ and Cd2+ transporter from the plant Arabidopsis thaliana, which is a member of the Heavy Metal ATPase family, was also expressed in C. reinhardtii. HMA4 full length and C-terminal tail expression strains were screened for Zn and Cd tolerance and uptake. Both sets of strains showed a significant increase in Cd and Zn tolerance and uptake compared to the wild type. Metal tolerance and uptake was compared between the genetically engineered C. reinhardtii strains and unicellular green algal strains that are naturally adapted to metal tolerance which were P. hussi, P. kessleri, and C. luteoviridis. Results showed significant increase in Zn and Cd tolerance and uptake in the natural strains compared to the engineered strains. Therefore in addition to genetically engineered strains, naturally adapted strains could also be used as tools for effective metal bioremediation and pollutant treatment.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 128-140).

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 6 uppsatser.

by Lai Mei Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 130-136). / Abstracts in English and Chinese. / Acknowledgment --- p.i / Abstract --- p.ii / 論文撮耍 --- p.v / List of Figures --- p.viii / List of Tables --- p.xi / Abbreviations --- p.xii / Table of content --- p.xiii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Facilitative glucose transporters --- p.1 / Chapter 1.1.1 --- Predicted Secondary structure of Glutl --- p.1 / Chapter 1.1.2 --- The tissue-specific distribution of glucose transporters --- p.2 / Chapter 1.2 --- Increase of glucose uptake in cancer cells --- p.5 / Chapter 1.3 --- Antisense oligonucleotide therapeutics --- p.7 / Chapter 1.3.1 --- Chemical modifications of oligonucleotides --- p.7 / Chapter 1.3.2 --- Cellular Uptake of Oligonucleotide --- p.11 / Chapter 1.3.3 --- Mechanism of action --- p.13 / Antisense-mediated RNA Cleavage --- p.14 / """Occupancy-only"" mediated mechanism" --- p.15 / Chapter 1.3.4 --- Antisense treatment in vivo --- p.17 / Chapter 1.4.5 --- Human Studies of Antisense Treatment --- p.18 / Chapter Chapter 2 --- Materials & Methods --- p.20 / Chapter 2.1 --- Materials --- p.20 / Chapter 2.2 --- Cell Culture --- p.21 / Chapter 2.2.1 --- Human colon adenocarcinoma cell Line (Caco-2) --- p.21 / Chapter 2.3 --- General Methodology for treatment of cells with antisense oligonucleotides --- p.22 / Chapter 2.3.1 --- Treatment of cells with oligonucleotides --- p.22 / Chapter 2.4 --- Cytotoxicity Assay --- p.23 / Chapter 2.4.1 --- MTT assay --- p.23 / Chapter 2.4.2 --- 3H-thymidine incorporation --- p.23 / Chapter 2.5 --- RNA extraction --- p.24 / Chapter 2.6 --- Competitive Reverse-transcription polymerase chain reaction (RT-PCR) of glucose transporters --- p.25 / Chapter 2.7 --- Measurement of 2-deoxy-D-glucose and Fructose transport --- p.27 / Chapter 2.8 --- Western blotting --- p.28 / Chapter 2.9 --- Flow cytometry --- p.30 / Chapter 2.9.1 --- Measurement of cellular accumulation of fluorophore-labeled oligonucleotide --- p.30 / Chapter 2.10 --- Design of antisense oligonucleotide --- p.31 / Chapter 2.11 --- ATP assay --- p.34 / Chapter 2.12 --- Animals studies --- p.35 / Chapter Chapter 3 --- Optimization of phosphorothioate antisense oligonucleotide delivery by Lipofectin --- p.36 / Chapter 3.1 --- Introduction --- p.36 / Chapter 3.2 --- Measurement of oligonucleotide uptake --- p.38 / Chapter 3.2.1 --- Lipofectin as a delivery system for the oligonucleotide uptake --- p.39 / Chapter 3.2.2 --- Effect of Lipofectin ratio on the oligonucleotide uptake --- p.41 / Chapter 3.2.3 --- Effect of oligonucleotide concentration on the oligonucleotide uptake --- p.41 / Chapter 3.2.4 --- Effect of incubation time on the oligonucleotide uptake --- p.44 / Chapter 3.2.5 --- Effect of oligonucleotide length on cellular uptake --- p.44 / Chapter 3.3 --- Effect of Lipofectin on cell viability --- p.47 / Chapter Chapter 4 --- In vitro effect of Antisense Oligonucleotides against glucose transporters on Caco-2 Cell --- p.49 / Chapter 4.1 --- Introduction --- p.49 / Chapter 4.2 --- Design of Antisense Oligonucleotides against Glucose Transporters gene --- p.50 / Chapter 4.3. --- Antisense effect of different regions of antisense oligonucleotide --- p.52 / Chapter 4.4 --- Antisense and Sense effect of oligonucleotide against start codon (G5 7015) on Caco-2 cells --- p.59 / Chapter 4.4.1 --- Effect of oligonucleotide to Lipofectin ratio on cell viability --- p.59 / Chapter 4.4.2 --- Dose-Response Study: effect of concentration of antisense - oligonucleotide on cell viability --- p.61 / Chapter 4.4.3 --- Effect of length´ؤof oligonucleotide on cell viability --- p.61 / Chapter 4.4.4 --- Time-Response Study: effect of antisense oligonucleotide on cell viability --- p.66 / Chapter 4.5 --- "The effect of antisense oligonucleotide against Glut1, Glut3 and Glut5 on cell viability of Caco-2 cells" --- p.70 / Chapter 4.6 --- Analysis of ATP content in Caco-2 cells by using antisense oligonucleotide flanking start codon (G5 7015) --- p.72 / Chapter 4.7 --- Effect of G5 7015 on HepG2 cells --- p.72 / Chapter Chapter 5 --- Effect of antisense oligonucleotides against Glut5 on mRNA and Protein levels of Glut5 gene --- p.76 / Chapter 5.1 --- Introduction --- p.76 / Chapter 5.2 --- RT-PCR of Glut isoform in Caco-2 cells --- p.77 / Chapter 5.3 --- Effect of antisense oligonucleotides against Glut 5 on mRNA level in Caco-2 cells --- p.77 / Chapter 5.3.1 --- Effect of oligonucleotides targeted different region of Glut5 gene on Glut5 message level --- p.77 / Chapter 5.3.2 --- Reduction in expression of mRNA level of Glut5 by using antisense oligonucleotides targeting start codon (G5 7015) --- p.81 / Chapter 5.3.3 --- Study of the dose and time dependence on inhibition of mRNA expression in G5 7015 treated Caco-2 cells --- p.83 / Chapter 5.3.4 --- Cross-Inhibition of antisense targeting glucose transporter isoforms --- p.83 / Chapter 5.4 --- Reduction in Glut5 protein level using G5 7015 antisense oligonucleotide --- p.86 / Chapter 5.5 --- Inhibition of Glut5 activity using G57015 oligonucleotide --- p.88 / Chapter 5.6 --- Inhibition of Glut5 mRNA level in vivo --- p.93 / Chapter Chapter 6 --- The possible role for Glucose Transporters in the Modification of Multidrug Resistance in Tumor cells --- p.95 / Chapter 6.1 --- Introduction --- p.95 / Chapter 6.2 --- Materials & Methods --- p.97 / Chapter 6.2.1 --- Cell culture --- p.97 / Chapter 6.2.2 --- Chemicals --- p.98 / Chapter 6.2.3 --- Measurement of doxorubicin uptake --- p.99 / Chapter 6.3 --- The expression of P-glycoprotein and Doxorubicin resistance of R-HepG2 cells --- p.99 / Chapter 6.4 --- Comparison of H3-2-deoxyglucose uptake between HepG2 and R-HepG2 cells --- p.99 / Chapter 6.5 --- Quantification of Glut1 and Glut3 expression by RT-PCR --- p.102 / Chapter 6.6 --- Comparison of doxorubicin between HepG2 and R-HepG2 cells cultured accumulation in glucose free medium --- p.104 / Chapter 6.7 --- The time course of doxorubicin accumulation in R-HepG2 cells culturing in glucose free medium --- p.106 / Chapter 6.8 --- "Cell viability of R-HepG2 cells after treatment of glucose transporter inhibitors, phloretin (PT), cytochalasin B (CB) and mitochondrial inhibitor，2,4-Dinitrophenol (DNP)" --- p.106 / Chapter 6.9 --- "Effect of glucose transporter inhibitors (PT, CB) and mitochondrial inhibitor (DNP) on doxorubicin accumulationin R-HepG2" --- p.110 / Chapter 6.10 --- Effect of antisense oligonucleotide against Glutl on doxorubicin accumulation in R-HepG2 cell --- p.113 / Chapter 6.11 --- "Analysis of ATP content and 3H-2-deoxy-D-glucose uptakein R-HepG2 after treatments of PT, CB and DNP" --- p.115 / Chapter Chapter 7 --- Discussion --- p.117 / Chapter 7.1 --- Antisense oligonucleotide against glucose transportersin Caco-2 cell --- p.117 / Chapter 7.2 --- Cellular uptake of oligonucleotide --- p.119 / Chapter 7.3 --- In vitro study of using antisense oligonucleotide against Glut5 --- p.121 / Chapter 7.4 --- In vivo study of using antisense oligonucleotide against Glut5 --- p.126 / Chapter 7.5 --- Possible role of inhibition of glucose transport in reversing P- gp --- p.127 / Chapter Chapter 8 --- References --- p.130

Chung Ka Wing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 151-162). / Abstracts in English and Chinese. / Contents --- p.i / Acknowledgements --- p.v / Abstract --- p.vi / 論文摘要 --- p.ix / List of Abbreviations --- p.xi / List of Figures --- p.xiii / List of Tables --- p.xv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Breast Cancer --- p.2 / Chapter 1.1.1 --- Incidence Rate of Breast Cancer --- p.2 / Chapter 1.1.2 --- Risk Factors Lead to Breast Cancer --- p.5 / Chapter 1.1.3 --- Conventional Treatments --- p.5 / Chapter 1.2 --- Relationship between Breast Cancer and Glucose Transporters --- p.7 / Chapter 1.2.1 --- Importance of Glucose and Fructose --- p.7 / Chapter 1.2.2 --- Facilitative Glucose Transporters (Gluts) and The Relationship with Breast Cancer --- p.7 / Chapter 1.3 --- Antisense Oligonucleotides --- p.13 / Chapter 1.3.1 --- Characteristics of Antisense Oligonucleotides --- p.13 / Chapter 1.3.2 --- Action Mechanism of Antisense Oligonucleotides --- p.15 / Chapter 1.3.3 --- Sequence Selection --- p.19 / Chapter 1.3.4 --- Chemical Modifications of Antisense Oligonucleotides --- p.20 / Chapter 1.3.5 --- Uptake and Delivery Means of Antisense Oligonucleotides --- p.24 / Chapter 1.4 --- Objectives of Present Study --- p.26 / Chapter Chapter 2 --- Materials and Methods --- p.31 / Chapter 2.1 --- Materials --- p.32 / Chapter 2.1.1 --- Cell Lines and Culture Medium --- p.32 / Chapter 2.1.2 --- Buffers and Reagents --- p.33 / Chapter 2.1.3 --- Reagents for Transfection --- p.34 / Chapter 2.1.4 --- Reagents for D-[U14C]-Fructose and 2-Deoxy-D-[l-3H] Glucose Uptake Assay --- p.35 / Chapter 2.1.5 --- Reagents for ATP Assay --- p.35 / Chapter 2.1.6 --- Reagents for RT-PCR --- p.36 / Chapter 2.1.6.1 --- Reagents for RNA Extraction --- p.36 / Chapter 2.1.6.2 --- Reagents for Reverse Transcription --- p.36 / Chapter 2.1.6.3 --- Reagents for Gel Electrophoresis --- p.37 / Chapter 2.1.7 --- Reagents for Real Time-PCR --- p.38 / Chapter 2.1.8 --- Reagents and Chemicals for Western Blotting --- p.39 / Chapter 2.1.8.1 --- Reagents for Protein Extraction --- p.39 / Chapter 2.1.8.2 --- Reagents for SDS-PAGE --- p.39 / Chapter 2.1.9 --- Reagents for Flow Cytometry --- p.42 / Chapter 2.1.10 --- In Vivo Study --- p.43 / Chapter 2.2 --- Methods --- p.44 / Chapter 2.2.1 --- Oligonucleotide Design --- p.44 / Chapter 2.2.2 --- Trypan Blue Exclusion Assay --- p.47 / Chapter 2.2.3 --- Transfection --- p.47 / Chapter 2.2.4 --- MTT Assay --- p.47 / Chapter 2.2.5 --- D-[U14C]-fructose and 2-deoxy-D-[l-3H] Glucose Uptake Assay --- p.48 / Chapter 2.2.6 --- Detection of Intracellular ATP Concentration --- p.49 / Chapter 2.2.7 --- Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.51 / Chapter 2.2.7.1 --- RNA Extraction by TRIzol Reagent --- p.51 / Chapter 2.2.7.2 --- Determination of RNA Concentration --- p.51 / Chapter 2.2.7.3 --- Reverse Transcription --- p.52 / Chapter 2.2.7.4 --- Polymerase Chain Reaction (PCR) --- p.52 / Chapter 2.2.8 --- Real-Time PCR --- p.55 / Chapter 2.2.8.1 --- Analysis of the Real-Time PCR Data --- p.57 / Chapter 2.2.9 --- Western Blot Analysis --- p.58 / Chapter 2.2.9.1 --- Protein Extraction --- p.58 / Chapter 2.2.9.2 --- Protein Concentration Determination --- p.58 / Chapter 2.2.9.3 --- Western Blotting --- p.60 / Chapter 2.2.10 --- Flow Cytometry --- p.62 / Chapter 2.2.10.1 --- Detection of Cell Cycle Pattern with PI --- p.62 / Chapter 2.2.10.2 --- Detection of Apoptosis with Annexin V/PI --- p.62 / Chapter 2.2.11 --- In Vivo Study --- p.63 / Chapter 2.2.11.1 --- Establishment of Tumor-Bearing Animal Model --- p.63 / Chapter 2.2.11.2 --- Treatment Schedule --- p.63 / Chapter 2.2.11.3 --- Toxicity of Antisense Oligonucleotides --- p.64 / Chapter Chapter 3 --- Results --- p.66 / Chapter 3.1 --- In Vitro Study --- p.67 / Chapter 3.1.1 --- Effect of Tamoxifen on MCF-7 cells and MDA-MB-231 cells --- p.67 / Chapter 3.1.2 --- Cytotoxicity of Antisense Oligonucleotides against Glut 5 on MCF-7 cells and MDA-MB-231 cells by MTT Assay --- p.69 / Chapter 3.1.3 --- Effect of Antisense Oligonucleotides against Glut 5 on Fructose and Glucose Uptake of MCF-7 cells and MDA-MB-231 cells by D-[U14C]-Fructose & 2-Deoxy-D-[l-3H] Glucose Uptake Assay --- p.77 / Chapter 3.1.4 --- Effect of Antisense Oligonucleotides against Glut 5 on Intracellular ATP Content of MCF-7 cells and MDA-MB-231 cells by ATP Assay --- p.81 / Chapter 3.1.5 --- Effect of Antisense Oligonucleotides against Glut 5 on Glut 5 RNA Expression of MCF-7 cells and MDA-MB-231 cells by RT-PCR and Real-Time PCR --- p.83 / Chapter 3.1.5.1 --- RT-PCR --- p.83 / Chapter 3.1.5.2 --- Real-Time PCR --- p.87 / Chapter 3.1.6 --- Effect of Antisense Oligonucleotides against Glut 5 on Glut 5 Protein Expression of MCF-7 cells and MDA-MB-231 cells by Western Blot Analysis --- p.89 / Chapter 3.1.7 --- "Effect of Antisense Oligonucleotides against Glut 5 on Change in Cell Cycle Pattern of MCF-7 cells and MDA-MB-231 cells by Flow Cytometry, Using PI Stainning" --- p.93 / Chapter 3.1.8 --- "Effect of Antisense Oligonucleotides against Glut 5 on Induction of Apoptosis of MCF-7 cells and MDA-MB-231 cells by Flow Cytometry, Using Annexin V-FITC Stainning" --- p.98 / Chapter 3.2 --- In Vivo Study --- p.101 / Chapter 3.2.1 --- Animal Model: Nude Mice --- p.101 / Chapter 3.2.2 --- Effect of Antisense Oligonucleotides against Glut 5 on the MCF-7 cells-Bearing Nude Mice --- p.101 / Chapter 3.2.2.1 --- Change of Weight of the Tumor-Bearing Nude Mice --- p.101 / Chapter 3.2.2.2 --- Tumor Growth Rate --- p.105 / Chapter 3.2.2.3 --- Glut 5 RNA Expression by Real-Time PCR --- p.109 / Chapter 3.2.2.4 --- Glut 5 RNA Expression by Western Blotting --- p.111 / Chapter 3.2.3 --- "Assessment of Side Effects of Antisense Oligonucleotides against Glut 5, by Measuring the Plasma Enzyme Level" --- p.113 / Chapter Chapter 4 --- Discussion --- p.118 / Chapter 4.1 --- Antisense Oligonucleotides against Glut 5 on Human Breast Cancer --- p.119 / Chapter 4.1.1 --- Antisense Oligonucleotides Strategy --- p.119 / Chapter 4.1.2 --- Role of Glut 5 in Breast Cancer --- p.123 / Chapter 4.1.3 --- Effects of Tamoxifen on MCF-7 and MDA-MB-231 --- p.126 / Chapter 4.2 --- In Vitro Study of Antisense Oligonucleotides against Glucose Transporter 5 on Breast Cancer Cells --- p.127 / Chapter 4.3 --- In Vivo Study of Antisense Oligonucleotides against Glucose Transporter 5 on Breast Cancer Cells --- p.135 / Chapter 4.3.1 --- Effects of Antisense Oligonucleotides against Glut 5 on Body Weight and Tumor Size --- p.137 / Chapter 4.3.2 --- Expression Level of Glut 5 of the Tumor --- p.138 / Chapter 4.3.3 --- Assessment of Side Effects of Antisense Oligonucleotides against Glut 5，by Measuring the Plasma Enzymes Level --- p.140 / Chapter 4.4 --- Possible Mechanism of Antisense Oligonucleotides against Glut 5 on Breast Cancer --- p.141 / Chapter Chapter 5 --- Future Prospectus and Conclusions --- p.143 / Chapter 5.1 --- Future Prospectus of Antisense Oligonucleotides --- p.144 / Chapter 5.1.1 --- Antisense Oligonucleotides and Treatment of Breast Cancer --- p.144 / Chapter 5.1.2 --- Role of Glut 5 in Breast Cancer --- p.147 / Chapter 5.2 --- Conclusions and Remarks --- p.148 / References --- p.151