Dissertation (Ph.D.) -- University of Texas Southwestern Medical Center at Dallas, 2008. / Vita. Bibliography: p.127-131

Thesis (Ph. D.)--West Virginia University, 2003. / Title from document title page. Document formatted into pages; contains viii, 166 p. : ill. (some col.). Includes abstract. Includes bibliographical references.

Thesis (Ph. D.)--West Virginia University, 2005. / Title from document title page. Document formatted into pages; contains ix, 226 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 204-212).

No description available.

by Keung Ka Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 163-184). / Abstracts in English and Chinese. / Abstract --- p.ii / 摘要 --- p.iv / Acknowledgements --- p.vi / Table of Contents --- p.vii / List of Abbreviations --- p.xiv / List of Tables --- p.xvi / List of Figures --- p.xvi / Chapter CHAPTER 1. --- INTRODUCTION / Chapter 1.1. --- Events happened after brain injury --- p.1 / Chapter 1.2. --- Glial cells --- p.3 / Chapter 1.2.1. --- Microglia --- p.4 / Chapter 1.2.2. --- Oligodendrocytes --- p.5 / Chapter 1.2.3. --- Astrocytes --- p.5 / Chapter 1.2.3.1. --- Uptake of neurotransmitters --- p.7 / Chapter 1.2.3.2. --- Maintenance of extracellular homeostasis --- p.8 / Chapter 1.2.3.3. --- Induction of blood-brain-barrier --- p.8 / Chapter 1.2.3.4. --- Guidance of migrating neurons during development --- p.9 / Chapter 1.2.3.5. --- Immunocompetent cells of the brain --- p.9 / Chapter 1.2.3.6. --- Contribution to astrogliosis --- p.10 / Chapter 1.3. --- Cytokines and astrogliosis --- p.11 / Chapter 1.3.1. --- IL-6 and astrogliosis --- p.12 / Chapter 1.3.2. --- IL-1 and astrogliosis --- p.13 / Chapter 1.3.3. --- IFN-γ and astrogliosis --- p.14 / Chapter 1.3.4. --- TNF-α and astrogliosis --- p.14 / Chapter 1.3.4.1. --- General properties of TNF-α --- p.15 / Chapter 1.3.4.2. --- TNF receptors (TNFRs) --- p.17 / Chapter 1.3.4.3. --- NFkB induction --- p.18 / Chapter 1.3.4.4. --- Intermediate early genes --- p.19 / Chapter 1.3.4.5. --- iNOS is the target of NFkB and AP-1 --- p.20 / Chapter 1.4. --- β-Adrenergic receptors (P-ARs) --- p.21 / Chapter 1.4.1. --- β-ARs and astrogliosis --- p.22 / Chapter 1.4.2. --- General properties of β-ARs --- p.23 / Chapter 1.4.3. --- Interactions between β-adrenergic mechanism and TNF-α --- p.24 / Chapter 1.5. --- Aims and scopes of the project --- p.25 / Chapter CHAPTER 2. --- MATERIALS & METHODS / Chapter 2.1. --- Materials --- p.29 / Chapter 2.1.1. --- Rats for astrocyte culture --- p.29 / Chapter 2.1.2. --- Cell culture materials --- p.29 / Chapter 2.1.2.1. --- Complete Dulbecco's Modified Eagle Medium:F12 (DF12) --- p.29 / Chapter 2.1.2.2. --- Phosphate buffered saline (PBS) --- p.30 / Chapter 2.1.3. --- Drugs preparation --- p.30 / Chapter 2.1.3.1. --- Recombinant cytokines --- p.30 / Chapter 2.1.3.2. --- Modulators of protein kinase A (PKA) --- p.30 / Chapter 2.1.3.3. --- Modulators of protein kinase C (PKC) --- p.31 / Chapter 2.1.3.4. --- β-Agonists and -antagonists --- p.31 / Chapter 2.1.3.5. --- Antibodies used in western blot analysis --- p.31 / Chapter 2.1.4. --- Reagents for cell proliferation determination --- p.32 / Chapter 2.1.5. --- Reagents for RNA isolation --- p.32 / Chapter 2.1.6. --- Reagents for reverse transcription-polymerase chain reaction (RT-PCR) --- p.32 / Chapter 2.1.7. --- Reagents for Electrophoresis --- p.33 / Chapter 2.1.8. --- Reagents and buffers for western blotting --- p.35 / Chapter 2.2. --- Methods --- p.36 / Chapter 2.2.1. --- Preparation of primary astrocytes --- p.36 / Chapter 2.2.2. --- Preparation of cells for assays --- p.36 / Chapter 2.2.3. --- Determination of cell proliferation --- p.36 / Chapter 2.2.3.1. --- [3H]-Thymidine incorporation assay --- p.37 / Chapter 2.2.3.2. --- MTT assay --- p.37 / Chapter 2.2.3.3. --- Data analysis --- p.38 / Chapter 2.2.4. --- Determination of RNA expression by RT-PCR analysis --- p.38 / Chapter 2.2.4.1. --- RNA extraction --- p.38 / Chapter 2.2.4.2. --- Spectrophotometric Quantitation of DNA and RNA --- p.38 / Chapter 2.2.4.3. --- RNA gel electrophoresis --- p.39 / Chapter 2.2.4.4. --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.39 / Chapter 2.2.4.5. --- Separation of PCR products by agarose gel electrophoresis --- p.40 / Chapter 2.2.4.6. --- Quantification of band density --- p.41 / Chapter 2.2.5. --- Determination of protein expression by Western blotting --- p.41 / Chapter 2.2.5.1. --- Total protein extraction --- p.41 / Chapter 2.2.5.2. --- Western blotting analysis --- p.42 / Chapter CHAPTER 3. --- RESULTS / Chapter 3.1. --- Effects of pro-inflammatory cytokines on astrocyte proliferation --- p.43 / Chapter 3.1.1. --- Effects of TNF-α on astrocyte proliferation --- p.44 / Chapter 3.1.2. --- Effects of TNF-R1 and -R2 antibodies on astrocyte proliferation --- p.47 / Chapter 3.1.3. --- Effects of other cytokines on astrocyte proliferation --- p.50 / Chapter 3.1.4. --- Comparisons of the effects of cytokines on astrocyte proliferation --- p.53 / Chapter 3.2. --- Effects of β-agonist and -antagonist on astrocyte proliferation --- p.55 / Chapter 3.3. --- Effects of TNF-α on the expression of TNFR and endogenous TNF-α in astrocytes --- p.60 / Chapter 3.3.1. --- Effects of TNF-α on the expression of TNF-R1 and -R2 in astrocytes --- p.60 / Chapter 3.3.1.1. --- Effects of TNF-α on the expression of TNF-R1 and -R2 mRNA --- p.60 / Chapter 3.3.1.2. --- TNFR subtypes involved in the TNF-α-induced TNF-R2 mRNA expression --- p.62 / Chapter 3.3.1.3. --- Signaling pathways of the TNF-α-induced TNF-R2 mRNA expression --- p.67 / Chapter 3.3.1.4. --- Effects of TNF-α on the expression of TNF-R1 and -R2 --- p.68 / Chapter 3.3.2. --- Effects of TNF-α on the expression of endogenous TNF-α in astrocytes --- p.73 / Chapter 3.3.2.1. --- Effects of TNF-α on the expression of TNF-α mRNA --- p.73 / Chapter 3.3.2.2. --- TNFR subtypes involved in the TNF-α-induced TNF-α mRNA expression --- p.73 / Chapter 3.3.2.3. --- Signaling pathways of the TNF-α-induced TNF-α mRNA expression --- p.74 / Chapter 3.3.2.4. --- Effects of other cytokines on the expression of TNF-α mRNA --- p.75 / Chapter 3.4. --- Effects of TNF-α on the expression of β1- and β2-AR in astrocytes --- p.85 / Chapter 3.4.1. --- Effects of TNF-α on the expression of β1- and β2-AR mRNA --- p.85 / Chapter 3.4.2. --- TNFR subtypes involved in the TNF-a-induced β1 and β2-AR mRNA expressions --- p.88 / Chapter 3.4.3. --- Signaling pathways of the TNF-α -induced β1- and β2-AR mRNA expressions --- p.88 / Chapter 3.4.4. --- Effects of TNF-α on the expression of β1- and β2-AR protein --- p.100 / Chapter 3.4.5. --- Effects of other cytokines on the expression of β1- and β2-AR mRNA --- p.100 / Chapter 3.5. --- Interactions between TNF-α and β-adrenergic mechanism in astrocytes --- p.107 / Chapter 3.5.1. --- Effects of β-agonists and -antagonists on the TNF-α-induced endogenous TNF-α expression in astrocytes --- p.107 / Chapter 3.5.1.1. --- Effects of ISO and PROP on the expression of TNF-α mRNA --- p.107 / Chapter 3.5.1.2. --- β-AR subtypes involved in the TNF-α-induced TNF-α mRNA expression --- p.108 / Chapter 3.5.2. --- Effects of β-agonists and -antagonists on the TNF-α-induced TNFRs expression in astrocytes --- p.112 / Chapter 3.5.2.1. --- Effects of ISO and PROP on the expression of TNFRs mRNA --- p.112 / Chapter 3.5.2.2. --- β-AR subtypes involved in the TNF-α-induced TNF-R2 mRNA expression --- p.115 / Chapter 3.6. --- Effects of TNF-α on the expression of transcription factors in astrocytes --- p.117 / Chapter 3.6.1. --- "Effects of TNF-α on c-fos, c-jun and NFKB/p50 expression" --- p.118 / Chapter 3.6.2. --- Effects of other cytokines on the expression of NFKB/p50 mRNA --- p.119 / Chapter 3.6.3. --- "TNFR subtypes involved in the TNF-α-induced c-fos, c-jun and NFKB/p50 mRNA expression" --- p.125 / Chapter 3.7. --- Effects of TNF-α on the expression of iNOS in astrocytes --- p.130 / Chapter 3.7.1. --- Effects ofTNF-α the expression of iNOS mRNA --- p.130 / Chapter 3.7.2. --- TNFR subtypes involved in the TNF-α-induced iNOS mRNA expression --- p.131 / Chapter 3.7.3. --- Signaling pathways of the TNF-α-induced iNOS mRNA expression --- p.136 / Chapter 3.7.4. --- Effects of other cytokines on the expression of iNOS mRNA --- p.139 / Chapter 3.7.5. --- Effects of β-agonists and -antagonists on the TNF-α-induced iNOS expression --- p.142 / Chapter 3.7.5.1. --- Effects of ISO and PROP on the expression of iNOS mRNA --- p.142 / Chapter 3.7.5.2. --- β-AR subtypes involved in the TNF-α-induced iNOS mRNA expression --- p.143 / Chapter CHAPTER 4. --- DISCUSSIONS & CONCLUSIONS / Chapter 4.1. --- Effects of TNF-α on astrocyte proliferation --- p.148 / Chapter 4.2. --- Roles of endogenous TNF-α and TNFR in astrocyte proliferation --- p.150 / Chapter 4.3. --- Interactions between TNF-α and β-adrenergic mechanism in astrocytes --- p.154 / Chapter 4.4. --- Induction of transcription factors by TNF-α in astrocytes --- p.157 / Chapter 4.5. --- Possible source of β-agonists --- p.159 / Chapter 4.6. --- Conclusions --- p.160 / REFERENCE --- p.163

Thesis (Ph.D. in Immunology) -- University of Colorado, 2005. / Typescript. Includes bibliographical references (leaves 190-212). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;

No description available.

The a of TNFa in title is the Greek alpha. / Thesis (MSc)--University of Stellenbosch, 2002. / ENGLISH ABSTRACT: Background: Cardiac hypertrophy is an adaptive process occurring in response to mechanical overload or tissue injury. The stimuli for cardiac hypertrophy are diverse and vary from increased afterload on the heart to cardiac remodeling in response to cytokines. Amongst others, obesity is characterized by excessive body weight resulting in metabolic disorders. This excess body weight necessitates an increased blood and oxygen delivery to the peripheral tissues, which is achieved by an elevated cardiac output. Total blood volume is also increased in the obese due to the increased tissue volume and vascularity. With time, the obesity induced increase in cardiac preload results in left ventricular hypertrophy and dilatation. Obesity is also associated with complications such as hypertension, insulin resistance and impaired glucose metabolism. In addition, adipose tissue has been implicated to contribute to elevated circulating TNFa levels in obesity and may contribute to the pathophysiology of the heart in obese individuals. The heart is a major cytokine-producing organ that generates amongst others tumor necrosis factor a (TNFa). TNFa is a proinflammatory cytokine, which acts to increase its own production, has cytotoxic and cytostatic effects on certain tumor cells and influences growth and differentiation in virtually all cell types including cardiomyocytes. Elevated levels of TNFa are detected peripherally in almost all forms of cardiac injury and in hypertrophic cardiomyopathy. These elevations are proposed to be deleterious to the heart, although an adaptive role for low levels of TNFa has been proposed. Aim: The aim of the study was to determine whether there is a correlation between obesity and serum, myocardial, and adipose tissue TNFa levels and cardiac hypertrophy. We also wished to determine whether the hearts from the obese animals functioned normally under normoxic conditions and whether they responded differently to ischaemia/reperfusion when compared with their concurrent controls. Materials and Methods: Male Sprague-Dawley rats (n=100) were fed a high caloric diet (HCD) containing 33% rat chow, 33% condensed milk, 7% sucrose and 27% water, or standard laboratory rat chow for 6-12 weeks. Food consumption, body weight gain, heart weight and tibia length were measured. Serum glucose, insulin and lipid levels were also determined. Hearts were excised and perfused on the isolated Working Heart perfusion apparatus and cardiac function was monitored and documented. Hearts were then subjected to 15 minutes of total global ischaemia at 370C, and reperfused for 30 minutes. Cardiac function was again documented. A separate series of hearts were freeze-clamped at different time points during the experimental protocol and stored in liquid nitrogen for the determination of myocardial TNFa and cGMP levels. Serum TNFa levels were determined after 12 weeks on the high caloric or normal/control diet. After 12 weeks on the diet myocardial TNFa levels of the HCD fed animals and their concurrent controls were determined before and during ischaemia. Adipose tissue and myocardial tissue TNFa levels were also determined after 6, 9 and 12 weeks on the respective diets. Myocardial cGMP levels were measured in the HCD fed rats and the control rats after 6, 9, and 12 weeks. These data were used as an indirect index to determine whether the myocardial NOcGMP pathway was activated in the normoxic hearts on the respective diets. Results: The body weight of the HCO fed animals was significantly higher compared with their respective controls after 12 weeks on the diet (459.9 ± 173.8 g and 271.5 ± 102.6 g respectively (p<0.05». The HCO fed animals also had heart weight to body weight ratios that were significantly greater compared with the controls (4.2 ± 0.1 mglg and 3.7 ± 0.1 mglg respectively (p<0.05». The plasma glucose levels of the HCO fed animals were higher than their respective controls (9.2 ± 0.3 mmoiII and 7.8 ± 0.3 mmoiII respectively (p<0.05)), but their insulin levels were similar (12.87 ± 1.02 IlIUlml and 12.42 ± 5.06 IlIU/ml). Plasma lipid profiles (plasma cholesterol, high density lipoprotein (HOL) cholesterol and plasma triacylglyceride (TAG)) were abnormal in the HCO fed animals compared with the control rats. Plasma TAG levels in the HCO fed animals were significantly higher compared with the control rats (0.664 ± 0.062 mmoiII and 0.503 ± 0.043 (p<0.05», while plasma cholesterol levels (1.794 ± 0.058 mmoIII and 2.082 ± 0.062 mmoiII (p<0.05» and HOL cholesterol levels were significantly lower (1.207 ± 0.031 mmoiII and 1.451 ± 0.050 mmoiII (p<0.05». Cardiac mechanical function was similar for both groups before ischaemia, but the percentage aortic output recovery was lower for the hearts from the HCO fed animals when compared with their controls (47.86 ± 7.87% and 66.67 ± 3.76 % respectively (p<0.05». Serum TNFa levels of the HCO fed animals were higher compared with the control animals (51.04 ± 5.14 AU and 31.46 ± 3.72 AU respectively (p<0.05», but myocardial TNFa levels remained lower in these animals (312.0 ± 44.7 pglgram ww and 571.4 ± 132.9 pg/gram ww respectively (p<0.05)). During ischaemia these myocardial TNFa levels increased above those of the controls (442.9 ± 12.4 pg/gram ww and 410.0 ± 12.5 pg/gram ww respectively (p<0.05)). The adipose tissue TNFa levels were significantly increased after 12 weeks on the high caloric diet compared with the control animals (4.4 ± 0.4 pg/gram ww and 2.5 ± 0.3 pg/gram ww respectively (p<0.05)). There was no significant difference in the myocardial cGMP levels of the HCD rats compared with the conrol rats after 6, 9 and 12 weeks. Conclusion: 1) The high caloric diet induced obesity, which lead to cardiac hypertrophy in this study. 2) There was a strong correlation between elevated adipose tissue and serum TNFa levels, and cardiac hypertrophy. 3) Elevated serum TNFa levels did not lead to activation of the myocardial NO-cGMP pathway in the normoxic hearts in this model. 4) The hypertrophied hearts from the HCD fed animals had poorer post-ischaemie myocardial functions than their concurrent controls. / AFRIKAANSE OPSOMMING: Agtergrond: Miokardiale hipertrofie is In aanpassing wat gebeur as In gevolg van meganiese oorbelading of weefsel beskadiging. Verskillende stimuli kan tot miokardiale hipertrofie aanleiding gee soos byvoorbeeld In verhoging in nalading, of miokardiale hermodellering in respons op sitokiene. Verhoging van voorbelading in vetsug mag ook tot hipertrofie aanleiding gee. Vetsug word gekenmerk deur In oormatige liggaamsmassa wat tot metaboliese versteurings lei. Die oormatige liggaamsmassa vereis In verhoging in bloed- en suurstofverskaffing aan die perifere weefsel wat deur In verhoging in die kardiale uitset vermag kan word. Die bloed volume van In vetsugtige individu word ook verhoog as gevolg van In verhoging in weefselvolume en vaskulariteit en met verloop van tyd induseer die verhoogde kardiale voorbelading linker ventrikulêre hipertrofie en dilatasie. Vetsug word ook met verskeie ander siekte toestande soos hipertensie, insulien weerstandigheid en versteurde glukose metabolisme, geassosieer. Vetweefsel dra ook by tot verhoging van tumor nekrose faktor alfa (TNFa) vlakke in die bloed, wat op sy beurt tot miokardiale hipertrofie mag bydra. TNFa is In proinflammatoriese sitokien wat sy eie produksie kan stimuleer. Dit het ook sitotoksiese en sitostatiese effekte op sekere tumor selle en kan groei en differensiasie in bykans alle seltipes, insluitende kardiomiosiete, stimuleer. Die hart kan ook TNFa produseer en verhoogde TNFa vlakke word feitlik in alle vorms van miokardiale besering en hipertrofiese kardiomiopatie waargeneem. Daar word voorgestel dat verhoogde TNFa vlakke vir die hart nadelig is, ten spyte van die vermoeding dat die sitokien In potensiële aanpassings rol by laer vlakke het. Doelstelling: Die doel van hierdie studie was om vas te stelof daar 'n verband tussen vetsug en serum, miokardiale en vetweefsel TNFa vlakke en miokardiale hipertrofie, bestaan. Ons het ook gepoog om te bepaal of harte van vetsugtige diere normaal funksioneer en of die response van sulke harte op isgemie-herperfusie van die van ooreenstemmende kontroles verskil. Materiaal en tegnieke: Manlike Sprague-Dawley rotte (n=100) is vir 6-12 weke op 'n hoë kalorie dieët (HKD) geplaas. Die HKD het uit 33% rotkos, 33% gekondenseerde melk, 7% sukrose en 27% water bestaan. Kontrole diere het standaard laboratorium rotkos ontvang. Voedselinname, liggaamsmassa toename, serum insulien, glukose en lipied vlakke is ook bepaal. Harte is geïsoleer en geperfuseer volgens die Werk Hart perfusie metode en hart funksie is gemonitor en gedokumenteer. Harte is vervolgens aan 15 minute globale isgemie by 3rC blootgestel en daarna weer vir 30 minute geherperfuseer waartydens hartfunksie weer gedokumenteer is. 'n Aparte groep harte is op spesifieke tydsintervalle gedurende die eksperimentele protokol gevriesklamp en in vloeibare stikstof gestoor vir die bepaling van miokardiale TNFa en sGMP vlakke. Serum TNFa vlakke is bepaal na 12 weke op die dieët. Na die diere 12 weke op die HKD was, is hierdie diere en hulooreenstemmende kontroles se miokardiale TNFa vlakke voor en na isgemie bepaal. Vetweefsel en miokardiale TNFa vlakke is ook onderskeidelik na 6, 9 en 12 weke bepaal. Miokardiale sGMP vlakke is in die HKD diere en in die kontrole diere na 6, 9 en 12 weke bepaal. sGMP vlakke is gebruik as 'n indirekte indeks van aktivering van die miokardiale NO-sGMP boodskapper pad. Resultate: Na 12 weke op die dieët was die liggaamsmassa van die HKD diere beduidend hoër in vergeleke met hulooreenstemmende kontroles (459.9 ± 173.8 g en 271.5 ± 102.6 g (p<0.05)). Die HKD diere se hart massa tot liggaam massa verhouding was ook beduidend hoër in vergelyking met die van kontroles (4.2 ± 0.1 mglg en 3.7 ± 0.1 mglg (p<0.05)). Alhoewel insulien vlakke dieselfde was (12.42 ± 5.06 j.lIU/ml en 12.87 ± 1.02 j.lIU/ml), was serum glukose vlakke van die HKD diere hoër as die van die ooreenstemmende kontroles (9.2 ± 0.3 mmoiii en 7.8 ± 0.3 mmoiii (p<0.05)). Plasma lipied profiele (HOL cholesterol, plasma cholesterol en trigliseriede) was abnormaal in die HKD diere. Plasma TAG vlakke in die HKD diere was beduidend hoër as die van die kontroles (0.664 ± 0.062 mmoiii en 0.503 ± 0.043 (p<0.05)), terwyl plasma cholesterol vlakke (1.794 ± 0.058 mmoiii en 2.082 ± 0.062 mmoiii (p<0.05)) en HOL cholesterol vlakke beduidend laer was (1.207 ± 0.031 mmoiii en 1.451 ± 0.050 mmoiii (p<0.05)). Miokardiale meganiese funksie was dieselfde vir beide groepe voor isgemie, maar die persentasie aorta omset herstel tydens herperfusie was laer in die HKD diere in vergelyking met die van kontrole diere (47.86 ±. 7.87% en 66.67 ± 3.76% (p<0.05)). Serum TNFa vlakke van die HKD diere was beduidend hoër as die van kontrole diere (51.04 ± 5.14 AU en 31.46 ± 3.72 AU (p<0.05)), maar miokardiale TNFa vlakke was laer (312.0 ± 44.7 pglgram nat gewig en 571.4 ± 132.9 pglgram nat gewig (p<0.05)). Die vetweefsel TNFa vlakke was ook beduidend verhoog na 12 weke op "n hoë kalorie dieët wanneer dit vergelyk word met die van kontrole diere (4.4 ± 0.4 pglgram nat gewig en 2.5 ± 0.3 pglgram nat gewig respektiewelik (p<0.05)). Daar was geenbeduidende verskille in die miocardiale vlakke van sGMP in die HKD diere in vergelyking met die kontroles na 6, 9 en 12 weke. Gevolgtrekkings: 1) "n Hoë kalorie dieët het in dié studie vetsug geïnduseer en tot miokardiale hipertrofie gelei. 2) Daar was "n positiewe korrelasie tussen verhoogde vetweefsel en serum TNFa vlakke, en miokardiale hipertrofie. 3) Verhoogde serum TNFa vlakke het nie tot die aktivering van die miokardiale NO-sGMP pad in hierdie model gelei nie. 4) Die hipertrofiese harte het tydens herperfusie ná isgemie swakker as hulooreenstemmende kontroles gefunksioneer.

published_or_final_version / abstract / Pathology / Master / Master of Philosophy

published_or_final_version / abstract / Surgery / Master / Master of Philosophy