Linus Pauling incorporated hemoglobin and a disease of the blood, sickle cell anemia, into many of his researches between the mid-1930s and mid-1970s. In the early 1930s Pauling became interested in organic chemistry and named hemoglobin as one of the first biochemical substances that he planned to analyze. In 1935 he published his first paper on hemoglobin, which determined the structure of the four hemes in hemoglobin. Pauling continued to study the structure of hemoglobin until the early 1950s when he proposed that it was an alpha-helix. In 1945 Pauling learned about sickle cell anemia and published an important paper in 1949 with Harvey A. Itano, S.J. Singer, and Ibert C. Wells titled "Sickle Cell Anemia, a Molecular Disease." Pauling investigated hemoglobin into the mid-1970s when he tried to find an orthomolecular therapy for sickle cell anemia. From the mid-1950s to early 1970s, Pauling also used sickle cell anemia to promote negative eugenics, point out the possible mutagenic effects caused by nuclear weapons testing, and propose an evolutionary theory. Additionally, in the final year of his life, Pauling wrote two forewords for books on sickle cell anemia, which were published in 1994, the year he died. Hemoglobin and sickle cell anemia can be considered a theme within Pauling's work. He often returned to normal and abnormal hemoglobin as his primary substance for examination, and his familiarity with hemoglobin and sickle cell anemia inspired new research. / Graduation date: 2004

Nucleic acids are an emerging class of therapeutics with the capacity to repair both DNA and RNA mutations in clinically relevant targets. We have used two approaches, mobile group II introns and Spliceosome Mediated RNA Trans-splicing (SMaRT), to correct β-globin mutations at the DNA and RNA levels respectively. We show that the group II intron inserts site-specifically into its DNA target, even when similar targets are available. Experiments transitioning this therapeutic into mammalian cell systems are then described. We also illustrate how SMaRT RNA repair can be used to correct β-globin mutations involved in sickle cell disease and some forms of β- thalassemia. We uncovered diverse repair efficiencies when targeting sickle cell versus β- thalassemia transcripts in mammalian cells. Possible reasons for this and how it might direct target choice for the SMaRT therapeutic approach are both discussed. The therapeutic molecule in SMaRT, a Pre-Trans-splicing Molecule or PTM, is also delivered via lentivirus to erythrocyte precursors cultured from the peripheral blood of sickle cell patients. Preliminary results from these experiments are discussed. / Dissertation

Sickle cell anemia is a hematological disorder that is caused by a single point mutation in the beta-globin chain of hemoglobin. It results in several complications related to the small and large vessels in patients with the disease. Large vessel complications include cerebral infarcts, which are observed in children under ten years old. The mechanism behind this complication is not completely understood. It is the goal of this project to begin to understand the role sickle erythrocytes may play in causing endothelial dysfunction as a precursor to sickle related complications. The hypothesis of this work is that exposure of large vessel endothelium to sickle erythrocytes causes an increase in endothelial permeability through loosening of adherens junctions. In the first goal of this work, bovine aortic endothelial cells (BAECs) are grown on coverslips and exposed to sickle erythrocytes for 5 minutes and either immediately fixed or incubated in 30 minutes and then fixed. Immunofluorescent studies labeling VE cadherin show changes in VE cadherin dynamics, suggesting sickle erythrocytes may be involved in this observation. Next, BAECs were grown on transwell inserts and exposed to sickle erythrocytes for 5 minutes. The erythrocytes are washed off and the BAEC are incubated with 10,000 MW dextran conjugated to lucifer yellow or FITC-BSA or to determine BAEC permeability. When dextran is used as the test molecule, endothelial permeability did not show a significant change from baseline. However, when BSA is used as the test molecule, increases in endothelial permeability are observed. Explanations into the differences between the transport mechanisms of the two molecules are discussed. These experiments show changes in VE cadherin localization due to sickle erythrocyte exposure. This may cause increases in endothelial permeability and an experimental model and preliminary studies are performed. This study provides potential mechanisms to explain the changes in VE cadherin localization and provide suggestions for further studies to test the effect of sickle erythrocytes on endothelial permeability. This work provides a strong foundation for continuing studies on the effects of sickle erythrocytes on endothelial dysfunction within the confines of sickle related complications.

Thesis (M. S. in Psychology)--Vanderbilt University, May 2006. / Title from title screen. Includes bibliographical references.

No description available.

Thesis (Ph.D)--University of Tennessee Health Science Center, 2008. / Title from title page screen (viewed on January 6, 2009). Research advisor: Ann K. Cashion, Ph.D. Document formatted into pages (x, 102 p. : ill.). Vita. Abstract. Includes bibliographical references (p. 81-88).

Thesis (M.S.)--University of Tennessee Health Sciences Center, 2005. / Spine title: Effects of sickle cell disease on growth of the craniofacial complexes. Appendices: leaves 162-414 Bibliography: leaves 145-161.

Thesis (Ph. D.)--Florida State University, 2003. / Advisor: Dr. Marcia L. Rosal, Florida State University, College of Visual Arts and Dance, Dept. of Art Education. Title and description from dissertation home page (viewed Jan. 31, 2005). Includes bibliographical references.

Thesis (PH.D.)--Michigan State University. Chemistry, 2008. / Title from PDF t.p. (viewed on Aug. 11, 2009) Includes bibliographical references. Also issued in print.

Thesis (Ph. D.)--Alliant International University, California School of Professional Psychology, San Francisco, 2005. / Includes bibliographical references (61-64) and abstract.