Thesis (Ph. D.)--University of Missouri-Columbia, 1996. / Typescript. Vita. Includes bibliographical references (leaves 279-302). Also available on the Internet.

Thesis (Ph. D.)--University of Missouri-Columbia, 1996. / Typescript. Vita. Includes bibliographical references (leaves 279-302). Also available on the Internet.

Thesis (M.S.)--University of Kentucky, 2007. / Title from document title page (viewed on June 28, 2007). Document formatted into pages; contains: vii, 74 p. Includes abstract and vita. Includes bibliographical references (p. 64-73).

Traditional meat production has major sustainability and ethical concerns. Cultivated meat helps to address these concerns by reducing the need for mass animal farming. Muscle progenitor cells (MPCs) harvested from skeletal muscle are a promising cell source for cultivated meat. While various protocols have been developed for MPC isolation, which protocol is best suited for the cultivated meat industry requires further investigation. Therefore, the purpose of this thesis was to optimize the MPC isolation technique to produce a pure myogenic cell population and provide the cultivated meat industry with standardized procedures for production. For these proof-of-concept experiments, skeletal muscles harvested from the hindlimb muscles of mice were used. Cells were isolated from the harvested muscle then subjected to one of three protocols for MPC enrichment: pre-plating, ice-cold treatment (ICT), or fluorescence activated cell sorting (FACS). The pre-plating and ICT protocols resulted in impure cell populations with few MPCs after one week in culture. Therefore, FACS using two cell-surface markers, NCAM and CD34, was employed as a more specific method for MPC sorting. CD34+NCAM1- cells grew quickly, however, unwanted cell types remained following FACS. In contrast, CD34+NCAM1+ cells had a consistent small, rounded shape and slow proliferation rate. These cells remained viable in culture for several months and had high Pax7 expression, indicating they were a pure population of myogenic cells. CD34+NCAM1+ cells maintained their capacity to differentiate after culturing for an extended period, demonstrating their potential use for cultivated meat production. The results of this study provide a better understanding of the differences between previously published MPC isolation techniques. Future studies will investigate the potential for CD34+NCAM1+ cells to be grown on a larger scale. These experiments provide insight into MPC populations that may exist in livestock species and will help to streamline the early stages of cultivated meat production. / Thesis / Master of Science (MSc) / Traditional meat production is associated with numerous challenges including animal welfare concerns, human health concerns, and harmful environmental consequences. The global population is predicted to reach 9.7 billion by 2050, emphasizing the importance of alternative food sources to meet the increased food demand. Cultivated meat is a promising new protein source, with the intended purpose of providing a sustainable food source with reduced ethical concerns compared to conventional meat. While there are several challenges to overcome throughout the production process, a major consideration in the early stages of cultivated meat production is cell sourcing. Muscle cells harvested from a tissue biopsy are one proposed starting cell source which has the potential to make up most of the tissue in cultivated meat products. This thesis aimed to improve upon previously published protocols used for muscle cell isolation and provide an optimized cell population for use in cultivated meat production. The cell sorting protocol described in this thesis provides a highly efficient technique for muscle cell purification and long-term growth. The resulting cell population has many characteristics that are pertinent to cultivated meat and may advance the early stages of production.

No / The use of non-cultivated plants in a daily diet based on local cuisines is potentially of considerable interest to nutritional scientists, because of the plants' role as local products and their potential as sources of novel nutraceuticals. In many Mediterranean regions these traditions are at risk of disappearing, hence the urgent need to study such knowledge systems. Accordingly, an ethnobotanical survey was carried out among the 850 inhabitants of the village of Castelmezzano, in central Lucania, which is located in the inland southern Italy. Seventy-five taxa of non-cultivated and semi-cultivated local food plants and mushrooms were documented, and uncommon food uses of a few species were reported for the first time. These include Bellavalia romana, Lepista nebularis and Onopordum illyricum. Most of the recorded non-cultivated food plants and mushrooms are cooked in oil or fat. Very few are consumed raw. This article discusses in detail the traditional culinary uses of these plants, their seasonality, ethnoecology, and their economic and nutritional potentials. The article also demonstrates how food agro-biodiversity is inextricably connected with cultural heritage.

Efficient in vitro generation of skeletal muscle tissue from cell lines is pivotal for the advancement of cultivated meat production. Here, we introduce a novel system employing a synthetic temperature-sensitive promoter (HSP70) carrying multiple heat shock elements (HSE), whose activity can be modulated by changes in temperature, enabling precise control of the expression of an exogenous master gene of myogenesis (MyoD) and a chromatin modifier (SMARCD3). To establish the optimal conditions, we screened different versions of temperature-inducible promoters (with 5, 15, or 45 HSE repetitions) in mammalian cell lines. Additionally, we explored the concurrent overexpression of exogenous heat shock factor 1 (HSF1), a multifaceted factor that enhances Hsp70 transcription, regulates the localization of binding partners, and modulates translation under stress. Our results demonstrate efficient induction of exogenous pro-myogenic factors with a 2-hour heat shock at 43°C. In the presence of exogenous HSF1, we observed a rapid initiation of the myogenic differentiation process and a significant acceleration in the expression of late myogenic markers (myogenin and sarcomeric myosin heavy chain), as well as the formation of skeletal muscle fibers in cells with intrinsic myogenic potential, such as L6 myoblasts. This approach allows for precise regulation of gene expression and cellular responses, leading to rapid and efficient production of mature muscle fibers without the need for expensive media formulations or genetic-based drug-inducible systems. This temperature-dependent cellular engineering method offers significant advantages, including accurate spatial and temporal control over cell myogenic maturation and scalability for large-scale production of differentiated skeletal muscle tissue for cultivated meat production.

Graduation date: 1999

　　Proliferative vitreoretinopathy (PVR) remains the major cause of failure of retinal detachment surgery. Retinal pigment epithelial (RPE) cells have been suggested to play a major role in the pathogenesis of PVR. Numerous studies have employed pharmacological means to modulate cellular activities in attempts to inhibit the process. Recent attempts using adjunctive therapy during PVR surgery that consisted of 5-fluorouracil (5-FU) and low molecular weight heparin showed some promise in preventing PVR but the concern is that prolonged 5-FU treatment may have a toxic effect. On the other hand, lectin from the edible mushroom Agaricus bisporus (ABL) was found to inhibit growth of RPE cells in a potent manner without apparent cytotoxicity. This lectin could be a candidate to modulate anomalous proliferation of RPE cells while the mechanism for the observed inhibition is unknown. 　　In our study, we investigated whether RPE cells treated with 5-FU or ABL would attenuate cellular proliferation, cell migration, cell adhesion and cell-mediated contraction rates. Further, we investigated if complementary inhibition for the above cellular activities could be obtained when RPE cells were treated with ABL after the short treatment using 5-FU. We also explored the possible mechanisms through which ABL inhibited RPE cell proliferation. 　　ARPE-19 and primary human RPE cells were treated with 5-FU or vehicle for 10 minutes. Cells were then maintained in culture medium supplemented with or without ABL. The rate of cellular proliferation was measured by a tetrazolium salt assay. Effects on cell adhesion were investigated through loading RPE cells onto the strips coated with collagen I or fibronectin. Cell migration was investigated using a scratch wound model. The effect on cell-mediated contraction was assessed using a free floating collagen I matrix. Cytotoxicity of 5-FU and ABL was determined by the live/dead assay. 　　To elucidate the mechanism through which ABL inhibited RPE cell proliferation, we investigated cell cycle distribution patterns using flow cytometry. Phosphorylation statuses of Erk, Jnk, p38, Akt as well as p53 and Cyclin D expression level were investigated by Western blotting. 　　Both 5-FU and ABL inhibited RPE cell proliferation. Only ABL promoted cell adhesion towards collagen I in hRPE3 cells. ABL was found to attenuate the rate of cell migration. Cell-mediated collagen gel contraction was attenuated by 5-FU only. Complementary inhibition in cellular proliferation and cell-mediated collagen gel contraction was observed when both 5-FU and ABL were applied. No significant cell death was observed after treatment with 5-FU, ABL or both. 　　ABL was found to reduce the amount of cells present at S phase. Akt and Erk were found to be hypo-phosphorylated and hyper-phosphorylated respectively after ABL treatment. The expression levels of phosphorylated-Jnk, phosphorylated-p38, p53, and Cyclin D1 were not altered when compared with the control. 　　These results showed that 5-FU and ABL complement with each other on inhibiting the wound healing activities of RPE cells in vitro without apparent cytotoxicity. They suggested a possible new treatment modality for PVR. ABL hypo-phosphorylated Akt and this observation is in line with the fact that ABL could attenuate cell proliferation. / published_or_final_version / Anatomy / Doctoral / Doctor of Philosophy

No description available.

Over the last century, much of the United State prairie pothole region has been converted to cultivated cropland. Concerns have been raised on government subsidized crop insurance effect on grassland to cropland conversion in the United States. Most grassland in the United State prairie pothole region are privately owned making a vivid understanding of landowners’ response to crop insurance subsidies important to facilitate effective design of prairie pothole conservation programs. The purpose of this study was to determine the extent of land use conversion with the removal of crop insurance subsidies in the United States prairie pothole region. Government subsidies on crop insurance was determined to have a measurable impact on Conservation Reserve Program land acreage within the United States. It was estimated that a 15% reduction in the net returns to cropland resulted in about 274,000 increase in the land acreage of the Conservation Reserve Program. In Conclusion, the removal of crop insurance subsidies was predicted to slow down but not stop the conversion of grassland to cropland in the United States prairie pothole region.