No description available.

No description available.

Ajonuma Louis Chukwuemeka. / "March 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 215-238). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Pup directed maternal licking and grooming (LG) increases with corticosterone (CORT) supplimentation (Rees et al 2004). Increases in LG lead to an attenuation of the adult offspring's HPA response to stress (Liu et aI1997). Similarly, Neonatal increases in glucocorticoids lead, in adulthood, to the same attenuation of the HPA stress response (Catalani et aI1993). We hypothesize that dams exhibiting increased LG will have increased circulating CORT, and this increase will be reflected in their offspring. This thesis characterizes the HPA hormone profile adrenocorticotropic hormone (ACTH), CORT & Corticosterone Binding Globulin (CBG) in High LG (H) and Low LG (L) litters, 5 days postpartum (P4). Furthermore pup plasma CORT levels are determined at (P) 3,4,6,10 & 14. Finally P10 Hand L LG ACTH, CORT & CBG will be assessed after stress. RESULTS: H compared to L LG dams have significantly increased plasma CORT (p=0.03). At P4, H LG offspring have significantly increased plasma CORT (p=0.03) and significantly decreased plasma ACTH (p=0.04) as compared to L LG offspring. Plasma CBG levels are significantly lower in H compared to L LG offspring (p=0.01) at the same age. Across the Stress Hyporesponsive Period (SHRP) H LG offspring had significantly increased plasma CORT (p= 0.00) compared to L LG offspring at P3. Challenged with a stressor at P10, H LG offspring have an exaggerated plasma CORT response (p=0.00). This data suggests increases in plasma CORT in the dams leads to increased CORT in the high offspring, contributing perhaps to a more mature stress response at P10. / Key word abbreviation: (1) CORT - CORTicosterone, (2) ACTH - AdrenoCorticoTropin releasing Hormone, (3) CBG - Corticosteroid Binding Globulin, (4) SHRP - Stress Hypo-Responsive Period, (5) P - Post-natal day, (6) HPA - Hypothalamic-Pituitary-Adrenal, (7) LG - Licking/Grooming, (8) ADX/OVX - ADrenalectomized/OVarectomized.

No description available.

INTRODUÇÃO: A relevância da expressão MYCN em neuroblastomas sem amplificação, diferentemente do seu aumento quando amplificado, permanece controverso. Neste trabalho, avaliou-se a relação do nível de expressão do transcrito MYCN em neuroblastoma não amplificado com os fatores clínicos e biológicos de prognóstico. MÉTODOS: Neste estudo observacional realizado entre janeiro de 2000 e dezembro de 2004, foram aferidos os valores do nível de expressão MYCN em 29 amostras tumorais, pela técnica RQ-PCR, no Laboratório de Biologia Tumoral da Fundação Pró-Sangue de São Paulo. Seus resultados foram analisados em relação à idade ao diagnóstico, ao estadiamento tumoral, ao grupo de risco, à ocorrência de recaída tumoral e de óbito. RESULTADOS: Foram nove crianças com idade 1 ano, os valores da expressão do transcrito MYCN variaram de 0,041 e 27,569, mediana 3,193. O estadiamento foi: quatro estádio 1; três estádio 2; oito estádio 3 e 14 crianças estádio 4. Entre 20 crianças com classificação patológica, 11 foram favoráveis e nove desfavoráveis. Considerando a mediana dos valores expressos a estratificação em grupos de risco com aumento da expressão foram: cinco crianças baixo risco; quatro risco intermediário e cinco alto risco. Grupos de risco sem aumento da expressão foram: duas crianças baixo risco; quatro risco intermediário e nove alto risco. Vinte e oito crianças obtiveram remissão completa e entre elas 14 apresentaram doença progressiva, sendo que sete morreram. As variáveis clínicas e biológicas não apresentaram freqüências diferentes entre os grupos de risco sem e com aumento de expressão. Entre os grupo alto risco e não alto-risco as variáveis idade, recaída tumoral e óbito apresentaram resultado com significado estatístico quando não se considerou o valor da expressão e quando não houve o seu aumento. Entre os grupos alto risco e nãoalto risco com aumento da expressão apenas a idade apresentou resultado com significado estatístico. CONCLUSÃO: Em crianças com estádio clínico não avançado o nível de expressão parece exercer uma relevância clínica, sugerindo um efeito protetor quanto menor for o aumento da expressão MYCN. / INTRODUTION: MYCN expression value in non-amplified neuroblastosmas remains a controversial issue. In order to add contributions to this field, children with nonamplified neuroblastomas were studied regarding their expression and correlation with clinical and other biological factors. METHODS: Twenty nine tumor samples obtained from non-consecutive patients admitted from January, 2000 through December, 2004, had their MYCN transcript expression levels evaluated according to the RQ-PCR assay, at the Tumoral Biology of Laboratory of the \"Fundação Pró -Sangue Hemocentro de São Paulo\", and compared to the following other factor: age at onset; tumor stage; risk - group; tumoral relapse rate and death. RESULTS: nine under one-year-old children and 20 over one-year-old children, with MYCN transcription expression level between 0.041 and 27.569, mean 3.193. Four children were stage 1, three were stage 2, stage 3 in eight and stage 4 in 14 children. In 20 patients with pathological classifications, 11 were favorable and nine unfavorable histology. Children whose expression level was above the mean were stratified as follows according to risk groups: five low-risk; four intermediate-risk and five high-risk patients. The ones whose expression level was under the mean were two low-risk, four intermediate-risk end nine high-risk patients. Twenty eight children achieved complete remission, with 14 recurrences, with seven deaths. The only factor associated to highly expressed MYCN patients was tumoral state. CONCLUSION: In children with non-advanced-stage disease low levels of expression might be a relevant favorable prognostic factor.

Liu, Xiaodong. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 135-155). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

by Mui Kin Tung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 97-113). / Chapter CHAPTER 1: --- INTRODUCTION --- p.1 / Chapter CHAPTER 2: --- LITERATURE REVIEW --- p.5 / Chapter 2.1 --- ION TRANSPORT SYSTEMS IN HUMAN ERYTHROCYTES --- p.6 / Chapter 2.1.1 --- "Sodium Pump (Na+,K+-ATPase)" --- p.6 / Chapter 2.1.2 --- Passive Sodium Transport Systems --- p.9 / Chapter 2.1.2.1 --- Sodium-potassium-chloride cotransport system --- p.9 / Chapter 2.1.2.2 --- Sodium-lithium Countertransport --- p.13 / Chapter 2.1.3 --- Ouabain- and Frusemide-Resistant Passive Effluxes --- p.17 / Chapter 2.2 --- ERYTHROCYTE SODIUM TRANSPORT SYSTEMS IN ESSENTIAL HYPERTENSION --- p.17 / Chapter 2.2.1 --- "Sodium Pump (Na+, K+-ATPase) in Essential Hypertension" --- p.18 / Chapter 2.2.2 --- Sodium-Potassium-Chloride Cotransport in Essential Hypertension --- p.20 / Chapter 2.2.3 --- Sodium-Lithium Countertransport in Essential Hypertension --- p.23 / Chapter 2.2.4 --- Passive Ion Fluxes in Essential Hypertension --- p.26 / Chapter 2.2.5 --- Intracellular Sodium Concentration in Essential Hypertension --- p.26 / Chapter 2.3 --- ERYTHROCYTE SODIUM TRANSPORT SYSTEMS IN DIABETES MELLITUS --- p.27 / Chapter CHAPTER 3: --- MATERIALS & METHODS --- p.29 / Chapter 3.1 --- MATERIALS --- p.30 / Chapter 3.1.1 --- Choline Wash Solution (CWS) --- p.30 / Chapter 3.1.2 --- Lithium Loading Solution --- p.31 / Chapter 3.1.3 --- Choline Wash Solution with Ouabain (CWS-O) --- p.31 / Chapter 3.1.4 --- Sodium Containing Medium (SCM) --- p.31 / Chapter 3.1.5 --- Sodium Free Medium (SFM) --- p.31 / Chapter 3.1.6 --- Sodium Free Medium with Bumetanide (SFM-B) --- p.32 / Chapter 3.1.7 --- Preservation Solution --- p.32 / Chapter 3.2 --- STUDY POPULATION --- p.32 / Chapter 3.2.1 --- Control Subjects --- p.35 / Chapter 3.2.2 --- Patients with Essential Hypertension --- p.35 / Chapter 3.2.3 --- Diabetic Patients --- p.35 / Chapter 3.3 --- DETERMINATION OF ERYTHROCYTE INTRACELLULAR SODIUM AND POTASSIUM CONCENTRATIONS (Naic/Kic --- p.36 / Chapter 3.3.1 --- Preparation of Erythrocytes --- p.36 / Chapter 3.3.2 --- Preparation of Haemolysates --- p.38 / Chapter 3.3.3 --- Determination of Sodium and Potassium Concentrations in Haemolysates --- p.38 / Chapter 3.3.4 --- Determination of Haemoglobin Concentration in Haemolysates --- p.38 / Chapter 3.3.5 --- Evaluation of Erythrocyte Intracellular Sodium and Potassium Concentrations --- p.39 / Chapter 3.4 --- DETERMINATION OF ERYTHROCYTE PASSIVE POTASSIUM EFFLUX --- p.39 / Chapter 3.4.1 --- Determination of Potassium Concentrations in Supernatant --- p.40 / Chapter 3.4.2 --- Evaluation of Passive Potassium Efflux --- p.40 / Chapter 3.5 --- DETERMINATION OF ERYTHROCYTE SODIUM-LITHIUM COUNTERTRANSPORT (SLC) AND LITHIUM-POTASSIUM COTRANSPORT (LPC) --- p.41 / Chapter 3.5.1 --- Lithium Loading --- p.42 / Chapter 3.5.2 --- Determination of Haematocrit --- p.42 / Chapter 3.5.3 --- Preparation of Haemolysates --- p.42 / Chapter 3.5.4 --- Determination of the Lithium Concentration in Haemolysates --- p.43 / Chapter 3.5.5 --- Determination of Lithium Efflux --- p.43 / Chapter 3.5.6 --- Evaluation of Lithium Efflux Rate --- p.43 / Chapter 3.5.7 --- Evaluation of Intracellular Lithium Concentration --- p.44 / Chapter 3.6 --- VALIDATION OF METHODOLOGY FOR DETERMINATION OF ERYTHROCYTE SODIUM TRANSPORT SYSTEMS --- p.45 / Chapter 3.6.1 --- Effect of Time Course of Lithium Efflux --- p.45 / Chapter 3.6.2 --- Intracellular Potassium Concentration and Its Effect on Ouabain- and Frusemide-Resistant Passive Potassium Efflux --- p.45 / Chapter 3.7 --- PRESERVATION OF ERYTHROCYTES FOR DETERMINATION OF SODIUM TRANSPORT SYSTEMS --- p.51 / Chapter 3.8 --- PRECISION OF THE METHOD --- p.51 / Chapter 3.9 --- STATISTICS --- p.52 / Chapter CHAPTER 4: --- RESULTS --- p.56 / Chapter 4.1 --- POPULATION CHARACTERISTICS --- p.57 / Chapter 4.2 --- ERYTHROCYTE INTRACELLULAR LITHIUM CONCENTRATIONS AFTER LITHIUM LOADING --- p.57 / Chapter 4.3 --- RELATIONSHIP BETWEEN ERYTHROCYTE ION TRANSPORT PARAMETERS AND OTHER VARIABLES --- p.58 / Chapter 4.4 --- ERYTHROCYTE SODIUM TRANSPORT SYSTEMS IN ESSENTIAL HYPERTENSION --- p.64 / Chapter 4.5 --- ERYTHROCYTE SODIUM TRANSPORT SYSTEMS IN PATIENTS WITH DIABETES MELLITUS --- p.64 / Chapter 4.5.1 --- NIDDM Patients without Hypertension --- p.64 / Chapter 4.5.2 --- NIDDM Patients with Hypertension --- p.65 / Chapter 4.5.3 --- NIDDM Patients with and without Hypertension --- p.65 / Chapter 4.6 --- ERYTHROCYTE SODIUM TRANSPORT SYSTEMS IN DIABETES MELLITUS PATIENTS WITH PROTEINURIA --- p.65 / Chapter 4.6.1 --- Clinical Features and Biochemistry Indices --- p.69 / Chapter 4.6.2 --- Ion Transport Systems and NIDDM Patients with Proteinuria --- p.69 / Chapter 4.7 --- EFFECTS OF TREATMENTS ON ERYTHROCYTE ION TRANSPORT SYSTEMS IN DIABETIC HYPERTENSIVE PATIENTS --- p.70 / Chapter 4.7.1 --- Effects of Diuretic Therapy --- p.70 / Chapter 4.7.2 --- Effects of Enalapril and Nifedipine Therapy --- p.74 / Chapter 4.7.3 --- Effects of Enalapril Therapy --- p.74 / Chapter 4.7.4 --- Effects of Nifedipine Therapy --- p.75 / Chapter 4.7.5 --- Comparison of the Effects of Enalapril and Nifedipine Therapy --- p.75 / Chapter CHAPTER 5: --- DISCUSSION --- p.81 / Chapter 5.1 --- SODIUM TRANSPORT IN ESSENTIAL HYPERTENSION --- p.82 / Chapter 5.1.1 --- Erythrocyte Sodium-Lithium Countertransport in Essential Hypertension --- p.82 / Chapter 5.1.2 --- Erythrocyte Sodium-Potassium Cotransport in Essential Hypertension --- p.86 / Chapter 5.1.3 --- Erythrocyte Intracellular Concentration of Sodiumin Essential Hypertension --- p.87 / Chapter 5.1.4 --- Erythrocyte Passive Potassium Efflux in Essential Hypertension --- p.90 / Chapter 5.2 --- SODIUM TRANSPORT SYSTEMS IN NON-INSULIN- DEPENDENT DIABETES MELLITUS (NIDDM) --- p.91 / Chapter 5.2.1 --- Sodium-Lithium Countertransport in Non-Insulin-Dependent Diabetes Mellitus --- p.91 / Chapter 5.2.2 --- Erythrocyte Lithium-Potassium Cotransport and Intracellular Sodium Concentration in Non-Insulin-Dependent Diabetes Mellitus --- p.93 / Chapter 5.3 --- EFFECT OF ANTIHYPERTENSIVE AGENTS ON ERYTHROCYTE SODIUM TRANSPORT SYSTEMS --- p.95 / REFERENCES --- p.98

To Wah Yuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 118-142). / Acknowledgements --- p.1 / Summary --- p.3 / List of Abbreviations --- p.7 / Chapter Part I --- Alpha2-HS glycoprotein: Identification and Characterization of the Jacalin Binding Properties --- p.8 / Chapter Chapter 1 --- Introduction --- p.9 / Chapter 1.1 --- Jackfruit and jacalin --- p.10 / Chapter 1.2 --- Biochemical and immunological properties of jacalin --- p.11 / Chapter 1.2.1 --- Molecular Weight of Jacalin --- p.11 / Chapter 1.2.2 --- Molecular structure of jacalin --- p.11 / Chapter 1.2.3 --- Specificity of jacalin to Thomsen-Fredenreich- antigen (T-antigen) --- p.13 / Chapter 1.2.4 --- The internal repeated sequence in the jacalin --- p.13 / Chapter 1.2.5 --- Jacalin-Bound Proteins (JBP) --- p.15 / Chapter 1.2.6 --- Interaction of jacalin to JBP --- p.15 / Chapter 1.2.7 --- Immunological properties of jacalin --- p.16 / Chapter 1.3 --- Application of jacalin in medical research --- p.17 / Chapter 1.4 --- Background knowledge of α2HSG --- p.18 / Chapter Chapter 2 --- Materials and Methods --- p.20 / Chapter 2.1 --- Design of experiment --- p.21 / Chapter 2.2 --- Identification of the Unknown JBP --- p.21 / Chapter 2.2.1 --- Sera --- p.21 / Chapter 2.2.2 --- Isolation of JBP by Affinity Chromatography --- p.22 / Chapter 2.2.3 --- Fast protein liquid chromatography (FPLC) of JBP --- p.22 / Chapter 2.2.4 --- Affinity chromatography with anti-human IgA column --- p.23 / Chapter 2.2.5 --- Preparation of non-IgA JBP fraction --- p.24 / Chapter 2.2.6 --- N-terminal sequencing of the non-IgA JBP fraction --- p.24 / Chapter 2.2.7 --- SDS-PAGE and immunoblot of gel filtration fractions --- p.25 / Chapter 2.2.8 --- ELISA of FPLC fractions of JBP --- p.26 / Chapter 2.2.9 --- Immunochemical analysis --- p.27 / Chapter 2.3 --- α2HSG: the property of jacalin binding --- p.28 / Chapter 2.3.1 --- α2HSG -jacalin binding curve and competitive ELISA --- p.28 / Chapter 2.3.2 --- Purification of jacalin-crude extract (JCE) --- p.28 / Chapter 2.3.3 --- Characterization of JCE and ASJ --- p.29 / Chapter 2.3.4 --- Comparison of jacalin from different sources for binding to α2HSG by competitive ELISA --- p.29 / Chapter Chapter 3 --- Results --- p.31 / Chapter 3.1 --- Identification of the unknown JBP --- p.32 / Chapter 3.1.1 --- Isolation and FPLC of JBP --- p.32 / Chapter 3.1.2 --- Identification of non-IgA JBP by anti-human IgA affinity column --- p.32 / Chapter 3.1.3 --- Identification of the known JBP in the FPLC fractionated JBP --- p.36 / Chapter 3.1.4 --- Characterization and confirmation of non-IgA JBP --- p.36 / Chapter 3.2 --- α2HSG: the property of jacalin binding --- p.42 / Chapter 3.2.1 --- Characterization of α2HSG-jacalin binding --- p.42 / Chapter 3.2.2 --- Characterization of the purified jacalin --- p.45 / Chapter 3.2.3 --- Comparison of different batches of jacalin to interact with α2HSG --- p.45 / Chapter Chapter 4 --- Discussion --- p.53 / Chapter Part II --- Jacalin-α2HSG binding: the Clinical Values --- p.57 / Chapter Chapter 5 --- Introduction --- p.58 / Chapter Chapter 6 --- Materials and Methods --- p.61 / Chapter 6.1 --- Preparation of IgA-specific jacalin (ASJ) by IgA-Sepharose 4B affinity column --- p.62 / Chapter 6.2 --- Preparation of JCE- and ASJ-Sepharose-4B affinity column --- p.62 / Chapter 6.3 --- Factors affecting the yield of α2HSG --- p.62 / Chapter 6.4 --- Miscellaneous methods --- p.63 / Chapter Chapter 7 --- Results and Discussion --- p.65 / Chapter Part III --- Application of Jacalin for Studying the Pathogenesis of IgA Nephropathy --- p.77 / Chapter Chapter 8 --- An Overview of IgA Nephropathy --- p.78 / Chapter 8.1 --- Clinical manifestation of IgA nephropathy --- p.79 / Chapter 8.2 --- Mesangial deposits in IgAN --- p.80 / Chapter 8.3 --- Human IgA system --- p.81 / Chapter 8.4 --- The role of circulating IgA in the pathogenesis of IgA nephropathy --- p.84 / Chapter 8.5 --- Pathogenesis of primary IgA nephropathy --- p.86 / Chapter 8.6 --- Interaction between circulatory IgA and fibronectin (FN) in primary IgAN --- p.87 / Chapter Chapter 9 --- Materials and Methods --- p.90 / Chapter 9.1 --- Design of experiment --- p.91 / Chapter 9.2 --- Sera --- p.91 / Chapter 9.3 --- Analysis of IgAl/IgA ratio in sera and JBP --- p.91 / Chapter 9.4 --- Purification and Fast protein liquid chromatography (FPLC) of jacalin-bound protein (JBP) --- p.92 / Chapter 9.5 --- Analysis of FPLC-fractionated JBP --- p.93 / Chapter 9.5.1 --- ELISA of IgA-containing immune complexes (IgA-IC) --- p.93 / Chapter 9.5.2 --- "Quantitative ELISA of IgA, K-IgAl, and λ-IgAl" --- p.94 / Chapter 9.5.3 --- "Measurement of sIgA,dIgA and IgA containing immune complex (IgA-IC)" --- p.95 / Chapter 9.5.4 --- SDS-PAGE analysis --- p.95 / Chapter 9.6 --- Statistics --- p.95 / Chapter Chapter 10 --- Results --- p.96 / Chapter 10.1 --- IgA1/IgA ratio of serum and JBP --- p.97 / Chapter 10.2 --- Isolation and FPLC of JBP --- p.97 / Chapter 10.3 --- SDS-PAGE analysis of FPLC fractionated JBP --- p.97 / Chapter 10.4 --- ELISA of the FPLC fractionated JBP --- p.99 / Chapter 10.4.1 --- "Distribution of IgA, secretory IgA (sIgA) and dimeric IgA (dIgA) in the FPLC fractions" --- p.99 / Chapter 10.4.2 --- Distribution of IgA containing immune complex (IgA-IC) in the FPLC fractions --- p.99 / Chapter 10.4.3 --- Quantitation of IgA --- p.106 / Chapter 10.4.4 --- "Quantitation of K-IgAl and λ-IgA1, and determination of k/λ ratio of IgAl" --- p.106 / Chapter 10.4.5 --- "Quantitation of sIgA,dIgA, and IgA-containing immune complex (IgA-IC)" --- p.109 / Chapter Chapter 11 --- Discussion --- p.112 / References --- p.118

by Nellie Yuk Fei Chung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 111-120). / Abstract also in Chinese. / Abstract --- p.i / List of Abbreviations --- p.iii / Acknowledgements --- p.iv / Publication of this study --- p.vi / Table of Contents --- p.vii / Chapter Chapter1: --- Introduction --- p.1 / Chapter 1.1 --- Multiple Myeloma --- p.2 / Chapter 1.2 --- The Problem --- p.2 / Chapter Chapter2: --- Literature Review --- p.5 / Chapter 2.1 --- Molecular Genetics of Multiple Myeloma --- p.6 / Chapter 2.1.1 --- Cytogenetics --- p.6 / Chapter 2.2 --- Alterations of Proto-Oncogenes --- p.9 / Chapter 2.2.1 --- c-myc --- p.9 / Chapter 2.2.2 --- Ras --- p.10 / Chapter 2.2.3 --- Bcl-2 and Related Protein --- p.10 / Chapter 2.3 --- Alteration of Tumor-Suppressor genes --- p.11 / Chapter 2.3.1 --- p53 Gene Mutations --- p.11 / Chapter 2.3.2 --- Retinoblastoma (Rb) Gene --- p.11 / Chapter 2.3.3 --- p16 and p15 Genes --- p.13 / Chapter Chapter3: --- DNA Methylation and Cancers --- p.14 / Chapter 3.1 --- Role of DNA Methylation --- p.15 / Chapter 3.2 --- CpG Islands --- p.15 / Chapter 3.3 --- Abnormalities of DNA Methylation in Neoplasia --- p.16 / Chapter 3.3.1 --- DNA Hypomethylation in Cancer --- p.16 / Chapter 3.3.2 --- DNA Methyltransferase Activity in Cancer --- p.17 / Chapter 3.4 --- Regional DNA Hypermethylation in Cancer --- p.17 / Chapter 3.4.1 --- p16 and p15 Genes in Solid Tumors --- p.18 / Chapter 3.4.2 --- The p16 and p15 Genes in Leukemia and other Hematopoietic Malignancies --- p.19 / Chapter 3.4.3 --- Retinoblastoma Gene --- p.20 / Chapter 3.5 --- Mechanism Underlying the DNA Methylation Changes --- p.21 / Chapter Chapter4: --- Background of Study --- p.23 / Chapter 4.1 --- Background of Study --- p.24 / Chapter 4.2 --- Project Objectives --- p.27 / Chapter Chapter5: --- Materials and Methods --- p.29 / Chapter 5.1 --- Patients Samples --- p.30 / Chapter 5.2 --- Normal Controls --- p.30 / Chapter 5.3 --- Storage of the Samples --- p.32 / Chapter 5.4 --- Materials --- p.32 / Chapter 5.4.1 --- Chemicals --- p.32 / Chapter 5.4.2 --- Primers --- p.33 / Chapter 5.4.3 --- Enzymes --- p.35 / Chapter 5.5 --- Methods --- p.35 / Chapter 5.5.1 --- Cloning of p16 and p15 Exon 1 Probes for Southern Analysis --- p.35 / Chapter 5.5.1.1 --- PCR Amplification of p16 and p15 exon1 Probes from Normal Blood DNA --- p.35 / Chapter 5.5.1.2 --- Recovery and Purification of p16 and p15 Exon 1 DNA Fragment --- p.36 / Chapter 5.5.1.3 --- Ligation --- p.37 / Chapter 5.5.1.4 --- Transformation --- p.37 / Chapter 5.5.1.5 --- Plating --- p.38 / Chapter 5.5.1.6 --- Screening of Recombinant Plasmid --- p.38 / Chapter 5.5.1.7 --- Confirmation of Cloned DNA by Sequencing --- p.42 / Chapter 5.5.2 --- DNA Extraction and Purification --- p.45 / Chapter 5.5.2.1 --- DNA Extraction from Bone Marrow Aspirate and Peripheral Blood --- p.45 / Chapter 5.5.2.2 --- Isolation of Plasmid DNA from Transformant Cutures --- p.46 / Chapter 5.5.2.3 --- Qualification and Quantification of DNA --- p.49 / Chapter 5.5.3 --- Detection of Hypermethylation by Southern Analysis --- p.50 / Chapter 5.5.3.1 --- Restriction Enzyme Digestion --- p.50 / Chapter 5.5.3.2 --- Agarose Gel Electrophoresis --- p.51 / Chapter 5.5.3.3 --- Southern Transfer --- p.51 / Chapter 5.5.3.4 --- Membrane Fixation --- p.51 / Chapter 5.5.3.5 --- Recovery and Purification of p16 and p15 Exon 1 Probes from Plasmid --- p.52 / Chapter 5.5.3.6 --- Probe Labeling --- p.54 / Chapter 5.5.3.7 --- Purification of Radioactive labeled DNA --- p.54 / Chapter 5.5.3.8 --- Southern Hybridization --- p.55 / Chapter 5.5.3.9 --- Post Hybridization --- p.55 / Chapter 5.5.3.10 --- Autoradiography --- p.56 / Chapter 5.5.4 --- Polymerase Chain Reaction-Single Strand Conformational Polymorphism Analysis (PCR-SSCP) --- p.56 / Chapter 5.5.4.1 --- 5'- end Radioactive Labeling of Primer --- p.56 / Chapter 5.5.4.2 --- Amplification of Target Sequence by PCR --- p.57 / Chapter 5.5.4.3 --- Non-denaturing Polyacrylamide Gel Electrophresis --- p.57 / Chapter 5.5.4.4 --- Direct DNA Sequence of PCR Products --- p.58 / Chapter 5.5.5 --- Prevention of Overall Contamination in PCR --- p.60 / Chapter 5.5.6 --- "Sensitivity, Specificity Controls" --- p.62 / Chapter Chapter6: --- Results --- p.64 / Chapter 6.1 --- Patient Characteristics --- p.65 / Chapter 6.1.1 --- General Patient Characteristics --- p.65 / Chapter 6.1.2 --- Clinical and Laboratory Features --- p.65 / Chapter 6.2 --- Southern Blot Analysis of p16/p15 and Rb --- p.79 / Chapter 6.2.1 --- Absence of Deletions or hypermethylationin Normal Controls --- p.79 / Chapter 6.2.2 --- Absence of Homozygous Deletions or Mutationsin p16/15 and Rb among all MM Patients --- p.79 / Chapter 6.2.3 --- Hypermethylation of p16 --- p.89 / Chapter 6.2.4 --- Hypermethylation of p15 --- p.92 / Chapter 6.3 --- Hypermethylation of p16/p15 and Clinico-pathologic Correlation --- p.94 / Chapter Chapter7: --- Discussion --- p.97 / Chapter 7.1 --- "Absence of Homozygous Deletions, Gene Rearrangements and Mutations in p16/p15 and Rb" --- p.98 / Chapter 7.2 --- Hypermethylation of p16/p15-An Alternative Way for Gene Inactivation --- p.100 / Chapter 7.2.1 --- Methylation of p15 Gene --- p.101 / Chapter 7.2.2 --- Methylation of 5'-CpG Island of p16/p15 and Lack of Gene Expression --- p.102 / Chapter 7.2.3 --- Comparison of Methylation Status of Primary Samples and Cell Lines in MM --- p.103 / Chapter 7.2.4 --- Progressive Gene Inactivation by Random Methylation Errors --- p.104 / Chapter 7.2.5 --- The Lack of Correlation of Tumor Contents Revealed by the Southern Analysis and Morphologic Assessment --- p.105 / Chapter 7.3 --- Knudson's Two-hit Model of Tumorigenesis --- p.106 / Chapter 7.4 --- Inverse Relationship of p16 and Rb --- p.107 / Chapter 7.5 --- Implications of Our Findings --- p.109 / Chapter 7.6 --- Future Studies --- p.109 / References --- p.111