Current Search: Kirchman, Paul A. (x)
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- Title
- Isolation of the methionine sulfoxide reductase B3 (MSRB3) gene from the red-eared slider, Trachemys scripta elegans.
- Creator
- Bishop, Eric, Kirchman, Paul A., Harriet L. Wilkes Honors College, Florida Atlantic University
- Abstract/Description
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Oxidative stress from reactive oxygen species (ROS) is hypothesized to be a primary cause of aging and aging-related diseases. For my thesis research, my advisor and I examined adaptation to oxidative stress in the red-eared slider, Trachemys scripta elegans, a turtle that goes long periods with low oxygen during hibernation, but then quickly re-acclimates to higher oxygen levels upon surfacing. This dramatic ischemia/reperfusion event should result in ROS formation, but the turtle appears to...
Show moreOxidative stress from reactive oxygen species (ROS) is hypothesized to be a primary cause of aging and aging-related diseases. For my thesis research, my advisor and I examined adaptation to oxidative stress in the red-eared slider, Trachemys scripta elegans, a turtle that goes long periods with low oxygen during hibernation, but then quickly re-acclimates to higher oxygen levels upon surfacing. This dramatic ischemia/reperfusion event should result in ROS formation, but the turtle appears to reduce oxidative stress. We aimed to isolate the gene for methionine sulfoxide reductase B3, a protein that alleviates oxidative stress by reconverting oxygen damaged methionine-R-sulfoxide to the amino acid methionine. We designed degenerate primers to amplify a conserved region of the MSRB gene to isolate a partial DNA sequence. We then used 5’ RACE PCR and vectorette library PCR to isolate much of the coding region of the gene from T. scripta.
Show less - Date Issued
- 2011
- PURL
- http://purl.flvc.org/fau/fd/FA00003560
- Format
- Document (PDF)
- Title
- Lowering oxidative stress with increased methionine content of mitochondrial aconitase.
- Creator
- Batlle, Jessica, Kirchman, Paul A., Harriet L. Wilkes Honors College, Florida Atlantic University
- Abstract/Description
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Oxidative stress occurs when reactive oxygen species (ROS), such as superoxide anion (O2' —), hydroxyl radical (HO.), and hydrogen peroxide (H2O2), build up to detrimental levels in the cell, and effects of oxidative stress are associated with aging. Mitochondria are the main site of oxidative stress, most likely due to the high level of ROS produced during respiration. Mitochondrial aconitase is an enzyme involved in respiration; due to the presence of an iron-sulfur cluster in the active...
Show moreOxidative stress occurs when reactive oxygen species (ROS), such as superoxide anion (O2' —), hydroxyl radical (HO.), and hydrogen peroxide (H2O2), build up to detrimental levels in the cell, and effects of oxidative stress are associated with aging. Mitochondria are the main site of oxidative stress, most likely due to the high level of ROS produced during respiration. Mitochondrial aconitase is an enzyme involved in respiration; due to the presence of an iron-sulfur cluster in the active site, it is a target for ROS. A plasmid containing the aconitase gene to be inserted into Saccharomyces cerevisiae was designed, and will be used in the future for the pop-in pop-out method; mitochondrial aconitase will be subjected to point mutations, replacing several leucine and isoleucine amino acid residues with methionine residues near the active site and on the surface of the enzyme. The amino acid methionine has a sulfur group, which also acts as a target for ROS and could prevent inactivation of the iron-sulfur cluster of aconitase.
Show less - Date Issued
- 2011
- PURL
- http://purl.flvc.org/fau/fd/FA00003557
- Format
- Document (PDF)
- Title
- Isolating the Manganese Superoxide Dismutase Gene (SOD2) from T. scripta"".
- Creator
- Sanchez, Jairo, Kirchman, Paul A., Chandrasekhar, Chitra, Harriet L. Wilkes Honors College, Florida Atlantic University
- Abstract/Description
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Reactive oxygen species (ROS) are molecular oxygen-derived molecules that are exceedingly reactive, often generated as free radical bi-products of mitochondrial respiration, which cause oxidative stress that leads to aging. To avoid the generation of ROS, aerobic organisms have antioxidant defense mechanisms that use enzymes such as Superoxide Dismutase (SOD) to convert the superoxide radicals into hydrogen peroxide and oxygen. Manganesecontaining SOD (Mn-SOD), a product of the SOD2 gene is...
Show moreReactive oxygen species (ROS) are molecular oxygen-derived molecules that are exceedingly reactive, often generated as free radical bi-products of mitochondrial respiration, which cause oxidative stress that leads to aging. To avoid the generation of ROS, aerobic organisms have antioxidant defense mechanisms that use enzymes such as Superoxide Dismutase (SOD) to convert the superoxide radicals into hydrogen peroxide and oxygen. Manganesecontaining SOD (Mn-SOD), a product of the SOD2 gene is found in all eukaryotic organisms in the mitochondrial matrix, including T. scripta (red-eared slider turtle). Extraordinarily, T. scripta can live without oxygen for long periods of time without experiencing adverse effects when oxygen is again available. The purpose of our experiment is to clone the T. scripta SOD2 gene, and test its activity. A partial fragment of the gene has been isolated by screening the cDNA library, and we are currently working on finding the remaining sequence.
Show less - Date Issued
- 2011
- PURL
- http://purl.flvc.org/fau/fd/FA00003605
- Format
- Document (PDF)
- Title
- Assaying Mutant Marine Bacteria for Lithium Extraction.
- Creator
- Alexander, Didier, Kirchman, Paul A., Wetterer, James, Harriet L. Wilkes Honors College, Florida Atlantic University
- Abstract/Description
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Lithium (Li) is used in many commercial products. With the recent development of a potent new lithium battery suitable for hybrid and plug-in electric cars, demand for lithium should soar. Seawater has low Li ion concentrations, and has not been considered a profitable commercial source of this element. For my thesis work, I attempted to develop a mutant strain of the marine bacteria Photobacterium damselae subsp. damselae capable of sequestering lithium from seawater. I expose the bacteria...
Show moreLithium (Li) is used in many commercial products. With the recent development of a potent new lithium battery suitable for hybrid and plug-in electric cars, demand for lithium should soar. Seawater has low Li ion concentrations, and has not been considered a profitable commercial source of this element. For my thesis work, I attempted to develop a mutant strain of the marine bacteria Photobacterium damselae subsp. damselae capable of sequestering lithium from seawater. I expose the bacteria to shortwave ultra violet (UV) light with the intent of obtaining a lithium dependent mutant. I was not successful at isolating such a mutant. However, I noted differences in colony size, between colonies grown in lithium media and sodium media, after UV treatment. Based on other research, the bacterial recovery methods of lithium and other metals from seawater remains a plausible option.
Show less - Date Issued
- 2011
- PURL
- http://purl.flvc.org/fau/fd/FA00003553
- Format
- Document (PDF)
- Title
- Analysis of Mutant Manganese-Superoxide Dismutase on the Lifespan of Saccharomyces cerevisiae"".
- Creator
- Salem, Sarah, Kirchman, Paul A., Wetterer, James, Harriet L. Wilkes Honors College, Florida Atlantic University
- Abstract/Description
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Previously isolated mutants of the superoxide dismutase 2 gene (SOD2), selected for increased activity in E. coli, were transformed into yeast cells in order to observe their effects on aging. Polymerase chain reaction and DNA sequencing were used to confirm that the yeast incorporated the mutant form of the gene. The strains with mutant forms of the SOD2 gene were observed to have decreased growth rates compared to the unmutated strain. Lifespan analyses were then conducted in order to see...
Show morePreviously isolated mutants of the superoxide dismutase 2 gene (SOD2), selected for increased activity in E. coli, were transformed into yeast cells in order to observe their effects on aging. Polymerase chain reaction and DNA sequencing were used to confirm that the yeast incorporated the mutant form of the gene. The strains with mutant forms of the SOD2 gene were observed to have decreased growth rates compared to the unmutated strain. Lifespan analyses were then conducted in order to see if yeast with mutant versions of SOD2 had lifespans that differed significantly from those with the unmutated forms of the gene. The mutant forms of SOD2 had no significant effect on the lifespan of yeast.
Show less - Date Issued
- 2011
- PURL
- http://purl.flvc.org/fau/fd/FA00003604
- Format
- Document (PDF)
- Title
- The Role of IL-17 in T-Cell Development.
- Creator
- Gopaldas, Manesh, Kirchman, Paul A., Chandrasekhar, Chitra, Harriet L. Wilkes Honors College, Florida Atlantic University
- Abstract/Description
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T-lymphocytes develop from bone marrow derived hematopoietic stem cells (HSCs) and mature in the thymus, where they participate in reciprocal signaling with thymic stromal cells. The thymic developmental stages are well characterized, but only a few intrathymic signals that influence the development of T-lymphocytes have been identified. Previous microarray experiments revealed interleukin-17A (IL-17A) and its receptor (IL-17RA) as a possible stromal-lymphoid signal. In this study, an IL-17RA...
Show moreT-lymphocytes develop from bone marrow derived hematopoietic stem cells (HSCs) and mature in the thymus, where they participate in reciprocal signaling with thymic stromal cells. The thymic developmental stages are well characterized, but only a few intrathymic signals that influence the development of T-lymphocytes have been identified. Previous microarray experiments revealed interleukin-17A (IL-17A) and its receptor (IL-17RA) as a possible stromal-lymphoid signal. In this study, an IL-17RA-/- knockout was used to determine whether the IL-17RA gene has a role in T-lymphocyte maturation. We made competitive bone marrow chimeras and analyzed the percentage of donor wildtype and mutant HSCs present in the bone marrow, and compared it to the percentage of a particular blood cell type that developed from these donor HSCs. We found that IL-17RA influences the maturation of T-lymphocytes, but does not affect the development of other immune cells such as B-lymphocytes, macrophages, and granulocytes.
Show less - Date Issued
- 2011
- PURL
- http://purl.flvc.org/fau/fd/FA00003578
- Format
- Document (PDF)