Thesis (Ph. D.)--Institute of Paper Chemistry, 1967. / Includes bibliographical references (leaves 83-87).

Thesis (Ph. D.)--Institute of Paper Chemistry, 1962. / Includes bibliographical references (p. 44).

Thesis (Ph. D.)--University of Washington, 2003. / Vita. Includes bibliographical references (leaves 109-113).

Critical to a cell's survival is its ability to deal with stress by making an appropriate response. This response often takes place in the cytoplasm, which is everything contained within the cell's plasma membrane that is not the nucleus. The cytoplasm is a dynamic environment and its ability to reorganize is essential to the cell's function. This dissertation presents a novel, previously undiscovered state of cytoplasm organization for the model system Schizosaccharomyces pombe, also known as fission yeast. Typically the fission yeast cytoplasm is a fluid-like environment in which endogenous lipid granules subject to thermal fluctuations, move freely as they explore their local surroundings through diffusion. When the cell is in a nutrient depleted environment it is exposed to the stress of advanced glucose starvation. As a result, we find that the cytoplasm undergoes drastic reorganization reminiscent of a phase transition; it is now a solid-like environment in which there is no visible motion. Lipid granules throughout the cell appear to be completely immobilized and are unable to move through the cytoplasm, despite the application of force through optical tweezers. We term this cytoplasmic state the cell frozen state. The cell frozen state is a physiological state, one that the cell can recover from with the addition of fresh nutrients. It is characterized by an anomalous diffusion exponent of [alpha] = 0.23 ± 0.01, which is a significant reduction from the anomalous diffusion exponent [alpha] = 0.66 ± 0.01 found for exponentially growing cells in which there is visible motion. To account for the cell wide immobilization of lipid granules, we hypothesize the formation of a polymer network all through the cytoplasm, and identify septins 1-3 as the most likely filament formers. In addition, we find there is an increase in the number of vacuoles in the cytoplasm during starvation, and propose a vacuole-septin model to describe the cytoplasm reorganization for the cell frozen state. / text

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Amino acids are important modulators of skeletal muscle metabolism, but their impact on glucose uptake by skeletal muscle remains unclear. To address the effect of an amino acid (AA) mixture consisting predominately of isoleucine on glucose uptake we first conducted a dose-response experiment, investigating how different concentrations of the AA mixture affect glucose uptake by isolated rat epitrochlearis muscle. In a subsequent experiment we examined how the AA mixture affects insulin-stimulated glucose uptake by isolated rat epitrochlearis muscle. It was found that the AA mixture with as little as 0.5 mM Ile increases [H3]2-deoxy-D-glucose (2-DG) uptake by 76% compared to basal glucose uptake. The AA mixtures with 1, 2 or 4 mM Ile provided no significant additional effect. Next we combined the AA mixture consisting of 2 mM Ile, 0.012 mM Cys, 0.006 mM Val and 0.014 mM Leu with physiological levels (75 μU/ml, sINS) and maximally-stimulating levels (2 mU/ml, mINS) of insulin. The AA mixture only, sINS and mINS significantly increased 2-DG uptake compared to basal by 63, 79 and 298%, respectively. When the AA mixture was combined with sINS and mINS 2-DG uptake was further increased significantly by 26 and 14%, respectively. Western blotting analysis revealed that compared to basal the AA mixture increased AS160 phosphorylation, while phosphorylation of Akt and mTOR did not change. Combining the AA mixture with sINS resulted in no additional phosphorylation compared to sINS alone. Interestingly, addition of the AA mixture to mINS resulted in increased phosphorylation of mTOR, Akt and AS160 compared to mINS alone. Our results suggest that certain AAs (1) increase glucose uptake in the absence of insulin and (2) augment insulin-stimulated glucose uptake in an additive manner. These effects on glucose uptake appear to be mediated via a molecular pathway that is partially independent from the canonical insulin signaling cascade. / text

No description available.

Objective: To characterize the association between longitudinal changes in fasting serum glucose and changes in flourodeoxyglucose Positron Emission Tomography (FDG PET) measurements of regional cerebral metabolic rate for glucose (rCMRgl) in brain regions preferentially affected by Alzheimer's disease (AD). A secondary objective was to investigate whether higher fasting serum glucose levels are associated with lower rCMRgl in younger adults within these same AD relevant brain areas. Methods: For the primary study, baseline, interim, and 4.4 ± 1.0-year follow-up fasting serum glucose and PET CMRgl were analyzed in 80 cognitively unimpaired, non-diabetic, 61.5 ± 5 year-old persons with a first-degree family history of AD, including 38 carriers and 42 non-carriers of the apolipoprotein E (APOE) ε 4 allele. An automated brain-mapping algorithm was used to characterize associations between changes in fasting serum glucose levels and changes in rCMRgl. Longitudinal changes in fasting serum glucose levels and their correlation with changes in six pre-selected neuropsychological test measures of memory, attention and processing speed were also assessed with linear regression. The secondary study included a cross sectional sample of 31 cognitively unimpaired, non-diabetic participants, 31.2 ±5.4 years of age. General linear model-based voxel-wise analyses were performed to examine the correlation between fasting serum glucose and rCMRgl. Results: In the primary study of older adults, average fasting serum glucose levels increased over longitudinal measurement, and changes in these levels were inversely associated with longitudinal CMRgl changes in the vicinity of brain regions preferentially affected by AD (p<0.05, corrected for multiple comparisons). Fasting serum glucose was also inversely associated with performance on a measure of visuospatial memory (p<0.05, corrected for multiple comparisons). In the younger sample, fasting serum glucose levels were inversely associated with rCMRgl in left frontal pole and right primary visual cortex regions (p<.05, corrected for multiple comparisons).Conclusions: In older adults, fasting serum glucose increases across time and is inversely related to rCMRgl in AD relevant regions and to visual memory test scores. This relationship between serum glucose and regional brain metabolism may begin in metabolically sensitive areas at a younger age.

The goal of this work was to characterize avian skeletal muscle (SKM) glucose and fatty acid uptake. English sparrows (Passer domesticus) were used for the following studies: 1. Characterization of glucose uptake, 2. Identification and localization of glucose transporters, 3. Characterization of free fatty acid uptake, and 4. Reciprocal inhibition of glucose and free fatty acids. The results are summarized as follows. Isolated SKM incubated for 60 minutes with insulin, IGF-1, caffeine or AICAR demonstrated no increase in glucose transport. Interestingly, uptake was decreased in the presence of incremental unlabeled glucose suggesting the presence of glucose transporters (GLUT) and by phloretin, an inhibitor of transport proteins, decreased transport. The SKM glycogen content was low, which is supportive of the observed minimal glucose uptake. These findings suggest that GLUT expression may differ in birds as compared to mammals. GLUT1 and GLUT3 gene expression, but not GLUT4, were found in all tissues examined and share a high degree of homology with published chicken sequences. In addition, GLUT3 and GLUT4 proteins were not detected, whereas GLUT1 protein was abundant in blood-tissue barriers. Sparrows have high plasma ketone body levels suggestive of a high rate of free fatty acid (FFA) oxidation. In vitro uptake of radiolabeled oleic acid (OA) was maximal at 60 minutes and competitively inhibited by unlabeled OA suggesting a facilitative process. Radiolabeled OA uptake was not increased by IGF-1, caffeine and AICAR, whereas insulin increased uptake at 60 minutes. Inhibitors of protein-mediated substrate transport increased OA uptake by 60 minutes (DIDS and phloretin) whereas a specific inhibitor of long chain FFA transport, sulfo-N-succinimidyl oleate, decreased its uptake at 2.5 min. In reciprocal inhibition studies, 20mM unlabeled glucose and OA inhibited the uptake of their radiolabeled counterparts. Glucose (20mM) significantly decreased labeled OA uptake 36% and 20mM OA significantly decreased labeled glucose transport by 49%. These data begin to elucidate why avian skeletal muscle may not take up glucose to an appreciable extent and further, why avian skeletal muscle is insulin resistant.