published_or_final_version / Physiology / Doctoral / Doctor of Philosophy

published_or_final_version / Pathology / Doctoral / Doctor of Philosophy

published_or_final_version / abstract / Medicine / Master / Master of Philosophy

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

No description available.

Leukemia is one of the most deadly diseases, responsible for the highest number of childhood cancer cases. Immunoglobulin-containing and proline-rich receptor-1 (IGPR-1) is a newly identified protein found to play an important role in human colon cancer and angiogenesis. The overall goal of this project was to assess IGPR-1 expression in leukemia cell lines and investigate its possible function in the NF-κB pathway, specifically the role of inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ) in the phosphorylation of IGPR-1. The NF-κB pathway, among others, plays a critical role in human-T-cell leukemia virus type 1 (HTLV-1) infected T-cells. Our preliminary results indicated that IGPR-1 is expressed in leukemia cell lines at variable levels. Further experiments demonstrated that IKKβ is involved in the phosphorylation of IGPR-1 as treatment of HEK-293 cells ectopically expressing IGPR-1 with an IKKβ inhibitor decreased IGPR-1 phosphorylation at Ser220. Likewise, cells treated with lipopolysaccharide (LPS), which is known to activate IKKβ, also stimulated the phosphorylation of IGPR-1 at Ser220. However, transfection of IGPR-1/HEK293 cells with Tax, an oncogene encoded by HTLV-1, decreased phosphorylation of IGPR-1 at Ser220. Taken together, our data indicates that activation of IKKβ in the NF-κB pathway stimulates phosphorylation of IGPR-1. / 2020-06-17T00:00:00Z

A Research Report submitted to the Faculty of Medicine, Department of Haematology, University of the Witwatersrand, Johannesburg for the Degree of Master of Medicine (in the branch Haematology). / WHSLYP2016

No direct stimulation of human peripheral blood mononuclear cells proliferation could be detected with incubation of various AB extracts including JAB80E70 in vitro. However, ex vivo assay performed using BALB/c mice orally fed with either water (control) or JAB80E70 for 28 days indicated for the first time that extract-treated groups significantly lowered the ex vivo mitogen-stimulated lymphocyte proliferation. / Our study started with the preparations of different AB extracts using various solvent systems with or without heating. Among all AB extracts tested, the extract JAB80E70 (extracted with 70% v/v ethanol at 80°C) showed the strongest selective suppression on the growth of human leukemia NB-4 and K-562 cells. The anti-leukemia effect of JAB80E70 was also confirmed in vivo using xenografted NB-4 bearing athymic nude mice model. / Phytochemical studies suggested that the selective anti-leukemia activity of JAB80E70-W-B-1 was retained in a relatively polar sub-fraction. One compound was isolated and identified as adenosine from JAB80E70-W-B-1. To the best of our knowledge, this is the first report of the presence of adenosine in AB, even though it exhibited no anti-leukemia activity in vitro. / The mushroom Agaricus blazei (AB) is traditionally used as a remedy against various cancers, infections and immune-related diseases. In the present study, the underlying mechanisms of the anti-tumor and immunomodulatory effects of AB extracts on human leukemia cells were systematically investigated. / With bioassay-guided fractionation, a sub-fraction (JAB80E70-W-B-1) with almost 5-fold improved selective cytotoxicity towards NB-4 cells was obtained. Both JAB80E70 and JAB80E70-W-B-1 could induce apoptosis characteristic DNA fragmentation and nucleosomes enrichment in NB-4 cells. An increase in the pro-apoptotic and a decrease in the anti-apoptotic Bcl-2 family proteins expression were observed in NB-4 cells treated with JAB80E70-W-B-1. JAB80E70-W-B-1 was also found to enhance activities of caspases 3, 8 and 9 in NB-4 cells. However, the activation of caspases 3 and 9 was not affected by the inhibition of caspase 8. Furthermore, JAB80E70-W-B-1 induced apoptosis in NB-4 cells was accompanied by a significant reduction in mitochondria membrane potential and telomerase activity. These results demonstrated that JAB80E70-W-B-1 induced apoptosis in NB-4 cells was dependent on caspase activity, and involved multiple molecular pathways. / Jiang, Jingjing. / Adviser: Lau Bik San Clara. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3454. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 224-250). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Chan, Hoi Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 206-220). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.ii / ABSTRACT --- p.viii / 撮要 --- p.xii / PUBLICATIONS --- p.xiv / TABLE OF CONTENTS --- p.xv / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- Hematopoiesis & Leukemia --- p.1 / Chapter 1.1.1 --- An Overview on Hematopoiesis Development --- p.1 / Chapter 1.1.2 --- Leukemia --- p.6 / Chapter 1.1.2.1 --- General Symptoms of Leukemia --- p.7 / Chapter 1.1.2.2 --- Classification of Leukemia --- p.8 / Chapter 1.1.2.3 --- Conventional Treatment against Leukemia --- p.15 / Chapter 1.1.2.4 --- Novel Approaches --- p.19 / Chapter 1.2 --- The Chinese Medicinal Herb-Banlangen (板藍根） --- p.24 / Chapter 1.2.1 --- An Overview on Natural Indigo Compounds Derived from Banlangen --- p.24 / Chapter 1.2.2 --- Tryptanthrin --- p.29 / Chapter 1.2.2.1 --- Anti-bacterial Activity of Tryptanthrin --- p.29 / Chapter 1.2.2.2 --- Anti-tumor Activity of Tryptanthrin --- p.31 / Chapter 1.2.2.3 --- Anti-inflammatory Activity of Tryptanthrin --- p.33 / Chapter 1.2.2.4 --- Cutting Edges of Tryptanthrin as a Drug --- p.34 / Chapter 1.2.2.5 --- Metabolism of Tryptanthrin --- p.35 / Chapter 1.3 --- Aims and Scopes of This Investigation --- p.37 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.39 / Chapter 2.1.1 --- Animals --- p.39 / Chapter 2.1.2 --- Cell Lines --- p.39 / Chapter 2.1.3 --- "Cell Culture Medium, Buffers and Other Reagents" --- p.41 / Chapter 2.1.4 --- Reagents for 3H-Thymidine Incorporation Assay --- p.45 / Chapter 2.1.5 --- Reagents and Buffers for Flow Cytometry --- p.46 / Chapter 2.1.6 --- Reagents for DNA Extraction --- p.49 / Chapter 2.1.7 --- Reagents for Measuring Caspase Activity --- p.50 / Chapter 2.1.8 --- Reagents for Total RNA Isolation --- p.53 / Chapter 2.1.9 --- Reagents and Buffers for Reversed Transcription-PCR --- p.54 / Chapter 2.1.10 --- Reagents and Buffers for Real Time-PCR --- p.59 / Chapter 2.1.11 --- Reagents and Buffers for Gel Electrophoresis of Nucleic Acids --- p.59 / Chapter 2.1.12 --- "Reagents, Buffers and Materials for Western Blot Analysis" --- p.61 / Chapter 2.2 --- Methods --- p.70 / Chapter 2.2.1 --- Culture of the Tumor Cell Lines --- p.70 / Chapter 2.2.2 --- "Isolation, Preparation and Culture of Mouse Peritoneal Macrophages" --- p.70 / Chapter 2.2.3 --- Determination of Cell Viability --- p.71 / Chapter 2.2.4 --- Determination of Cell Proliferation by [3H]-TdR Incorporation Assay --- p.72 / Chapter 2.2.5 --- Determination of Anti-leukemia Activity In Vivo --- p.73 / Chapter 2.2.6 --- Analysis of Cell Cycle Profile/DNA Content by Flow Cytometry --- p.74 / Chapter 2.2.7 --- Measurement of Apoptosis --- p.75 / Chapter 2.2.8 --- Determination of the Mitochondrial Membrane Potential --- p.77 / Chapter 2.2.9 --- Measurement of Caspase Activity --- p.78 / Chapter 2.2.10 --- Study of Intracellular Accumulation of Reactive Oxygen Species --- p.79 / Chapter 2.2.11 --- Gene Expression Study --- p.80 / Chapter 2.2.12 --- Protein Expression Study --- p.83 / Chapter 2.2.13 --- Measurement of Cell Differentiation --- p.87 / Chapter CHAPTER 3: --- STUDIES ON THE ANTI-PROLIFERATIVE EFFECT OF TRYPTANTHRIN AND INDIRUBIN-3'-OXIME ON MYELOID LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.90 / Chapter 3.2 --- Results --- p.94 / Chapter 3.2.1 --- Effects of Indirubin-3'-oxime and Tryptanthrin on the Proliferation of Myeloid Leukemia Cell Lines of Human and Murine Origins In Vitro --- p.94 / Chapter 3.2.2 --- Kinetic and Reversibility Studies of the Anti-proliferative Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.108 / Chapter 3.2.3 --- Cytotoxic Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.113 / Chapter 3.2.4 --- Cytotoxicity of Tryptanthrin on Non-Cancer Cell Line and Primary Myeloid Cells In Vitro --- p.115 / Chapter 3.2.5 --- Effects of Tryptanthrin on the Cell Cycle Profile of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.118 / Chapter 3.2.6 --- Effects of Tryptanthrin on the Expression of Cell Cycle Related Genes in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.123 / Chapter 3.2.7 --- Expression of CDK-inhibitors in Tryptanthrin-treated Murine Myeloid Leukemia WEHI-3B JCS Cells --- p.126 / Chapter 3.2.8 --- Effects of Tryptanthrin on the In Vivo Tumorigenicity of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.128 / Chapter 3.2.9 --- In Vivo Anti-tumor Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.130 / Chapter 3.3 --- Discussion --- p.132 / Chapter CHAPTER 4: --- STUDIES ON THE APOPTOSIS-INDUCING EFFECT OF TRYPTANTHRIN ON MURINE MYELOMONOCYTIC LEUKEMIA WEHI-3B JCS CELLS / Chapter 4.1 --- Introduction --- p.139 / Chapter 4.2 --- Results --- p.143 / Chapter 4.2.1 --- Induction of DNA Fragmentation by Tryptanthrin in the Murine Myelomonocytic Leukemia WEHI-3B Cells In Vitro --- p.143 / Chapter 4.2.2 --- Induction of Phosphatidylserine Externalization by Tryptanthrin in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.145 / Chapter 4.2.3 --- Change of Mitochondrial Membrane Potential of Tryptanthrin- treated Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.147 / Chapter 4.2.4 --- Induction of Caspase Activity in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.150 / Chapter 4.2.5 --- Induction of Reactive Oxygen Species in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.155 / Chapter 4.2.6 --- Expression of Bcl-2 Family Proteins in the Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.160 / Chapter 4.2.7 --- Effects of Tryptanthrin on the mRNA Expression of Bcl-2 Family Proteins in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.163 / Chapter 4.2.8 --- Expression of Fas and Fas Ligand Proteins in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.167 / Chapter 4.2.9 --- Expression of Pro-Apoptotic Protein in Tryptanthrin- treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.170 / Chapter 4.2 --- Discussion --- p.173 / Chapter CHAPTER 5: --- STUDIES ON THE DIFFERENTIATION-INDUCING ABILITY OF TRYPTANTHRIN ON MURINE MYELOMONOCYTIC LEUKEMIA WEHI-3B JCS CELLS / Chapter 5.1 --- Introduction --- p.184 / Chapter 5.2 --- Results --- p.186 / Chapter 5.2.1 --- Morphological Studies on Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.186 / Chapter 5.2.2 --- Effects of Tryptanthrin on the Cell Size and Granularity of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.189 / Chapter 5.2.3 --- Effects of Tryptanthrin on Induction of NBT-reducing Activity in the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.191 / Chapter 5.3 --- Discussion --- p.195 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.198 / REFERENCES --- p.206

Purpose: Pediatric dentists could serve a role in care of children with leukemia. Oral sequelae of cancer therapies are well documented. The purpose of this study is to assess the feasibility and effectiveness of the pediatric dentist in the care of patients with leukemia. Methods: Pediatric Hematology and Oncology at Virginia Commonwealth University was educated on the proposed protocol and administered a questionnaire to assess feasibility of implementing prevention education by the pediatric dentist. Patients were randomized into two groups at diagnosis: one receiving current oral health protocol and those receiving one-on-one prevention education with the pediatric dentist at three points during treatment. Data was collected through clinical intraoral examination and salivary sample. Results: All respondents reported this would address a known problem for patients. They agreed it is feasible and would be a valuable addition to care of these patients. Subject recruitment is ongoing for the effectiveness portion of the study. Conclusion: The addition of the pediatric dentist to the pediatric oncology care team is warranted and feasible.