Thesis (M.S.)--The University of North Carolina at Greensboro, 2010. / Directed by Michael McIntosh; submitted to the Dept. of Nutrition. Title from PDF t.p. (viewed Jul. 14, 2010). Includes bibliographical references (p. 42-50).

Thesis (M.A.)--University of North Carolina at Chapel Hill, 2004. / Includes bibliographical references (leaves 37-40). Also available online (PDF file) by a subscription to the set or by purchasing the individual file.

Thesis (Ph. D.)--University of Chicago, Committee on Human Nutrition and Nutritional Biology, August 2002. / Includes bibliographical references. Also available on the Internet.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1968. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of North Carolina at Greensboro, 2006. / Title from PDF title page screen. Advisor: Ron Morrison; submitted to the School of Human Environmental Sciences. Includes bibliographical references.

Thesis (Ph. D.)--University of North Carolina at Greensboro, 2006. / Title from PDF title page screen. Advisor: Michael K. McIntosh; submitted to the School of Health and Human Performance. Includes bibliographical references (p. 133-151).

Thesis (M.A.)--University of North Carolina at Chapel Hill, 2004. / Includes bibliographical references (leaves 37-40).

Impact of the chromatin remodeller SMARCAD1 on murine intestinal intraepithelial lymphocyte and white adipose tissue biology. Chromatin remodelling factors use the energy of ATP hydrolysis to drive the movement of and/or affect molecular changes to the nucleosome. One such factor, SMARCAD1 (SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A containing DEAD/H box 1), has been previously shown to restore heterochromatin at the replication fork in vitro. This project aimed to assess the impact of SMARCAD1 on mammalian biology, utilising an animal model in which the catalytic ATPase domain of murine SMARCAD1 had been deleted using Cre/lox technology. Preliminary results had implicated SMARCAD1 in adaptive-immunity and white adipose tissue biology, and SMARCAD1 expression in these tissues/cells was confirmed by tissue-panel western blot. This project therefore aimed to build on these results to understand better the impact of SMARCAD1 on adaptive immune development and white adipose tissue biology. In addition, fewer than expected viable Smarcad1-/- homozygous offspring were produced during Smarcad1+/- x +/- matings, which both confirmed the observation from a previous knockout model of Smarcad1, and limited the number of knockout animals available for this study. Investigation of systemic B- and T-cells in the bone marrow, thymus and spleen had previously suggested there was no significant defect in adaptive immune development in Smarcad1-/- mice, however a tissue-specific and age-related loss of intra-epithelial (IEL) T-lymphocytes was found in the small intestine by flow cytometry. Analysis by qPCR of duodenal RNA suggested that differentiation rather than inflammation may underpin any loss-of-IEL phenotype, although further examination of cell-proliferation and crypt/villus anatomy by EdU incorporation and immunofluorescence revealed no overt cell-anatomical or proliferative difference in the knockout mice. The requirement for large numbers of aged mice made further investigation of the intestinal IEL phenotype logistically prohibitive. The reduction of epididymal white adipose tissue (eWAT) size had also been observed in male Smarcad1-/- mice, and serum from these mice showed elevated triglyceride (TG) and free fatty acids (FFA) levels. Transcriptomic analysis by RNA-seq of whole-WAT revealed an elevation in macrophage-related markers in knockout mice, which was confirmed by flow cytometry. As a number of reports have implicated SMARCAD1 in stem cell biology, putative adipose stem cells were isolated from +/+ and -/- mice by FACS and used for adipogenic differentiation assays ex-vivo In parallel, mouse embryonic fibroblasts from +/- and -/- mice were also assayed for adipogenic differentiation. While no significant differences in adipogenesis were observed, Smarcad1-/- mice challenged with a (60%) high fat diet did show increased weight gain over +/+ mice, and measurements of adipocyte size and cell cycle/cell proliferation analysis suggested hyperplasia rather than defects in adipogenesis may drive any WAT-related pathology in these mice.

Leptin, a 16kDa protein, is secreted primarily by adipose tissue. Its effects are pleiotropic, with known roles in body mass regulation, haematopoiesis, immune function and reproduction. Studies of the leptin system in females were undertaken to complement existing animal and human studies. Serum leptin levels were found to increase in the luteal phase of the menstrual cycle raising the possibility of a role for leptin at the time of implantation. RT-PCR studies showed that leptin receptors were expressed in the human endometrium throughout the menstrual cycle. In these preliminary studies, these receptors appeared to be functional in response to leptin; increasing proliferation and decreasing TNF-a production in cultured endometrial cells. This implies that leptin has a local effect at the level of the endometrium. Many cytokines are bound to proteins in serum; a proportion of this binding can be attributed to a soluble cytokine receptor. Soluble receptors and binding proteins have a variety of roles, acting as scavengers, carriers, agonists and antagonists. In order to investigate this phenomenon in the leptin system, a radio-receptor assay was developed to measure leptin-binding activity (LBA). The leptin receptor has an extracellular domain that is common to all isoforms and therefore, LBA may also reflect the binding parameters of cell surface receptors. Serum leptin levels of LBA were found to be low at birth, high in early childhood, to fall steadily through puberty and remain at the post-pubertal levels throughout adult life. This suggests that leptin and LBA has an important role in the initiation of puberty. There was no significant variation in LBA during the menstrual cycle. Several single nucleotide polymorphisms (SNP) exist in the leptin receptor gene. Studies of two of these SNPs, GLN223ARG and LYS656ASN, present in the extracellular domain of the receptor were undertaken to assess if genetic changes are associated with differences in phenotype or effect ligand binding. Homozygosity of the G allele of GLN223ARG is associated with lower fat mass, BMI and leptin levels in postmenopausal Caucasian females. This polymorphism changes the binding characteristics of the receptor, with a higher LBA being associated with homozygosity of the G allele. This suggests that the actual binding of leptin to its receptor may be an important factor in the regulation of body weight and adiposity.

In the past 30 years the prevalence of obesity has almost trebled resulting in an increased incidence of type 2 diabetes mellitus (T2DM) and other co-morbidities. Visceral adipose tissue is believed to play a vital role in these conditions, but underlying mechanisms remain unclear. A close association exists between obesity, diabetes and oxidative stress, resulting in increased reactive oxygen species formation. The experiments in this thesis address this by searching for possible biochemical changes which may be specific for the onset of obesity related T2DM, as well as looking for genetic alterations at molecular and gene expression levels. This thesis also explored various techniques such as polymerase chain reaction (PCR), colorimetric assays and real-time RT-PCR. The aim was to investigate the role of adipose tissue in obesity and T2DM, focusing on markers of oxidative stress and gene expression in human visceral adipose tissue from subjects categorised as lean, obese and obese with T2DM. This cross-sectional study measured two markers of oxidative stress, two markers of DNA damage, gene expression analysis and identification of genes associated with T2DM and obesity. Specific gene sequencing was carried out on the glutathione reductase gene to determine possible gene variants. Results showed a paradoxical decrease in adipose markers of oxidative stress in subjects with obesity and T2DM. There appeared to be a protective mechanism in these subjects, displaying reduced levels of oxidative stress compared to other groups. This could be due to a significant proportion of these subjects being on ACE inhibitor and statin therapy, which may be confounding results and minimising the effects of the oxidative burden. Additionally, the same subjects showed an increased expression of the glutathione reductase gene. It is difficult to conclude if the decreased levels of oxidative stress in these subjects were a result of the increased glutathione reductase expression in the visceral adipose tissue or if there remains an unseen factor influencing the dramatic expression change seen in this group of subjects. No glutathione reductase gene variants were identified in these samples. This analysis highlighted that within this sample set, the impact of oxidative stress is in fact reversible as the antioxidant capacity in these subjects is evident, and in combination with correct drug therapy it may be possible to combat oxidative burden and reduce the subsequent damage inflicted upon the cells.