Thesis (Ph. D.)--University of Wisconsin--Madison, 1956. / Typescript. Abstracted in Dissertation abstracts, v. 16 (1956) no. 11, p. 2021. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 95-96).

Thesis (M.S.)--University of Wisconsin--Madison, 1966. / eContent provider-neutral record in process. Description based on print version record. Bibliography: l. 42-43.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1951. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 62-65).

Thesis (M.S.)--University of Wisconsin--Madison, 1949. / Typescript. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 40-44).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1960. / Typescript. Vita. Parts of Section I are reprinted from Biochimica et biophysica acta. Includes bibliographical references.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1953. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 57-58).

Age-related changes in the pH of the medium, fatty acid composition and biosynthesis of fatty acids in various subcellular fractions have been studied in submerged cultures of P. chrysogenum harvested at pH's 7.2, 4.8, 4.4 and 3.8. 14C - labeled lauric and myristic acids were each incubated with cell fractions prepared by sonification and fractional centrifugation. Products were analyzed by thin layer chromatography, gas liquid scintillation spectrometry. Conversion of labeled precursors into longer chain saturated and unsaturated fatty acids was employed as a measure of the activity of the various cell fractions.It was found that between pH 4.8 and 4.4 of the age of the culture a period of maximum fatty acid synthesis occured at the time when glucose was being taken up to a greatest extent from the medium and when cells start rapidly increasing in growth. This correlated to the increased rate of de novo synthesis in the soluble cytoplasmic fraction.The fermentation products, formic, acetic, proprionic, butyric and gluconic acid, produced at different time points of the age of the culture, accounted for the drop in the pH of the medium.

The synthesis of lipids and long chain fatty acids in young and aging cultures of Penicillium chrysogenum was studied by identifying the intracellular location of radioactively labeled intermediates in the vegetative mycelium and identifying the lipids and fatty acids into which the label from 1-14C-palmitic acid was incorporated.Previous work has indicated that the pH of the growth medium of submerged cultures of Penicillium chrysogenum dropped from pH 7.4 in newly inoculated cultures to 3.2-3.8 during the 20-40 hour growth period. Young cultures were defined as those harvested before the pH began to drop and aging cultures were those harvested after the pH stabilized at near 3.4. Changes in the fatty acid composition of various cell fractions of the mycelium harvested as the pH of the medium declined suggested that a change in utilization of lipids employed in the synthesis of structural components in the cell may have occurred as a result of a shift in fatty acid metabolism as the culture aged.Young and aging cultures were incubated with l-14C-palmitic acid and harvested after 2, 10, 60, and 120 minutes. Samples of mycelia from each harvest were examined by light and electron microscopy and were prepared for autoradiography. In addition lipids were extracted from sonified mycelial samples, analyzed for total lipids, lipid classes, and phospholipid components by thin layer chromatography. Fatty acids were identified and quantified by gas liquid chromatography with percent distribution of label in fatty acids determined by liquid scintillation spectrometry. Results of biochemical analysis of some of the major lipid components were compared with electron microscope autoradiographs and related to changes in the location of labeled fatty acid in cell organelles or cell parts as the cells aged.In young cultures 70% of the radioactive label was recovered in phospholipids while about 15% was recovered in the free fatty acid component. By contrast, in aging cultures approximately 20% of the label was recovered in the phospholipids and 80f was recovered from the free fatty acid component. Electron autoradiographic data tend to support these biochemical findings in that numerous grains occur over the membranal components of the young cells and over cytoplasmic areas of lipid depots in aging cells. Phospholipids differed markedly also with large amounts of an unidentified phospholipid type found in aging cells not seen in young cells.Light and electron microscope observations of hyphal cells showed significant alterations in cell morphology over the forty hour growth period. Young cells were long, slender, with dense cytoplasm and thin cell walls. As the culture aged, cells became progressively shorter, thicker and more clubby in appearance with prominent lipid inclusions. The vacuolar lipid depots of aging cells were ,determined to be composed of free fatty acids with 20% of label being incorpoated into C20 and C21 long chain fatty acids. The synthesis of fatty acid chains longerthan C18 has not been previously reported in intact cultures of Penicillium chrysogenum.The lipid metabolism of young cells of Penicillium chrysogenum differed from that of aging cells in many aspects. Young cells incorporated label from precursor palmitic acid into membranes. However, as the cells aged, lipids were diverted to storage. Young and aging cells differed in the amounts and composition of total lipids, lipid classes, phospholipid components and fatty acids. Differences in morphology between young and aging cells could be demonstrated by light and electron microscopy. These structural changes paralleled the biochemical changes indicating a functional dissimilarity existed between the young and aging cells.

Thesis (Ph. D.)--University of Wisconsin, 1964. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Bibliography: leaves 79-85.

Results of previous research on whole-cell cultures of Penicillium chrysogenum have suggested that acetyl CoA, without being converted to malonyl CoA, supplies the two carbon units for the elongation of palmitic acid. The purpose of this study was to determine the mode of elongation of 1-t4C palmityl CoA by a 20,000 x g mitochondrial pellet from P. chrysogenum.Acetyl CoA or malonyl CoA was incubated with radioactivelylabeled palmityl CoA for 20 minutes. Avidin was added to some experimental reaction mixtures. The resulting fatty acids were saponified, extracted with hexane, methylated with diazomethane, and purified by thin layer chromatography. The methyl esters were separated and identified by gas-liquid chromatography. The radioactivity of each methyl ester was determined by liquid scintillation spectrometry.Elongation of palmityl CoA was observed in the presence of acetyl CoA, but not in the presence of malonyl CoA. The addition avidin produced a greater proportion of short-chained fatty acidsthe expense of palmitic acid, but did not decrease the percentage of long-chained fatty acids produced.A high proportion of label was recovered in the C18:3 fatty acid, linolenic acid. This suggested that two pathways of linolenic acid synthesis may be operating in this organism.Methods for detection and control of cancer encompass a large area of today's research. Recent use of granulomas as a model for such detection and control may be a promising field, especially for monitoring tumor antigens and immune responses. These granuloma systems are increasingly becoming vehicles in the study of tumor immunology. Although granulomas may be induced naturally by means of foreign bodies i.e. viral, fungal, or bacterial agents, new methods are being established to produce artificial granuloma systems. These systems include chemical or foreign body implantations followed by tumor vaccine challenges. The research presented here involved the use of a golf ball-induced granuloma for the purpose of establishment of a detection system for immune responses. The use of a golf ball-induced granuloma provided a closed system for monitoring cell-mediated and humoral responses to tumor antigens. Immune responses were monitored by means of hematocrits (packed blood cell counts), white blood cell differential counts, and electrophoretic results.Hematocrit results indicated no great immune response to the closed vaccine injected granulomasystems. Observations made on differential white blood cell counts indicated decreasing neutrophil/lymphocyte ratios for cellular immune responses. Electrophoretic results for granuloma fluids indicated decreases in albumin levels concurrent with increases in peak two,and complete loss of peak three following vaccination. Responses to tumor specific antigens in the form of cell-mediated immune responses are indicated by the results presented in this research. Utilization of the golf ball-induced granuloma system provided a means of separating the cell-mediated and humoral immune responses.Tumor specific antigens elicited various immune responses and provide hope for future identification of tumors by this method. Future development and utilization of the golf ball-induced granuloma system may be potential means of monitoring cell-mediated immune responses to tumor malignancies.