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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
11

The initiation of autoimmune reactions in anticonvulsant drug-induced hypersensitivity: The role of cytochrome P450 enzymes

Kinobe, R. Unknown Date (has links)
No description available.
12

INVESTIGATION OF ARSENIC (AS) AND LEAD (PB) MIXTURE DEVELOPMENTAL TOXICITY

Keturah Gayle Kiper (12789119) 27 July 2023 (has links)
<p>   </p> <p>Environmental toxicants such arsenic (As) and lead (Pb) are chemicals that enter the environment and can result in adverse health effects humans, especially during development. This dissertation work evaluated As and Pb to determine if developmental toxicity significantly changes at lethal and sub-lethal mixture concentrations using the zebrafish model. Joint action models were applied to survival data to determine the type of interaction. Metal exposures were from 1-120 hours post fertilization (hpf). As concentrations were 0-7,500 ppb. Pb concentrations were 0–100,000 ppb. The LC25, LC50, and LC75 values at 120 hpf from single metal exposures were used to select mixture concentrations for modeling. The survival data indicated an additive effect occurred at lethal concentrations. </p> <p>The impact of the mixture on behavior, morphology, and gene expression was then evaluated at sub-lethal concentrations of 10 and 100 ppb As and Pb individually or in mixtures. Data was analyzed with a repeated measures ANOVA (behavior) or an ANOVA (morphology and qPCR) with the least significant difference test (α=0.05). Zebrafish larvae exposed to 10 ppb As exhibited hyperactivity in all dark phases for the distance moved, time moving, and velocity, while those exposed to 10 ppb Pb only showed an increase in distance moved and velocity in the first dark phase. The 10 ppb mixture was found to have an intermediate impact with increased time moving in all dark phases and increased distance moved and velocity only in the first dark phase. In contrast, hyperactivity was observed only in the 100 ppb As and 100 ppb mixture treatment in the last two dark phases for time spent moving. No significant behavioral alterations occurred in the 100 ppb Pb treatment. A decrease in mean brain length and brain length ratio to the total length in the 10 ppb mixture was observed with no significant morphology changes observed for head length, head width, or total length. Alternatively, measurements of cerebral vasculature morphology in the mesencephalon (midbrain) and cerebellum (hindbrain) uncovered decreased total vascularization at 72 hpf (exposure 1-72 hpf) in both brain regions. This decrease occurred in all treatment groups in the mesencephalon and in all treatment groups, except the 100 ppb Pb and 10 ppb As treatment groups in the cerebellum. In addition, decreased sprouting and branching occurred in the mesencephalon, while only decreased branching was measured in the cerebellum. The 10 ppb Pb treatment group showed unique perturbations in several cerebral vasculature endpoints evaluated, which was also observed in a specific gene expression alteration pattern different from the other treatment groups. To identify molecular changes associated with these changes, expression of genes related to angiogenesis and vasculogenesis (i.e., <em>vegfaa, wnt7aa,</em> and<em> lrp1aa</em>) and genes associated with tight junctions (i.e., <em>cldn5a</em> and <em>cldn5b</em>) were assessed at 72 hpf. Increased <em>cldn5b</em> expression was detected in all treatment groups, while <em>cldn5a </em>was increased in only the 10 ppb Pb treatment group. In addition, <em>wnt7aa</em> was only decreased in the 10 ppb Pb treatment group. Alternatively, <em>vegfaa</em> was increased in the 100 ppb As and 100 ppb mixture treatment groups and no changes were detected for <em>lrp1aa</em>. In summary, cerebral vascular toxicity outcomes in the 10 ppb mixture treatment were primarily driven by changes in the 10 ppb Pb treatment group, while perturbations in the 100 ppb mixture treatment group aligned with the 100 ppb As alterations. In addition, the non-linear dose response for 10 and 100 ppb Pb treatment groups agree with observations in prior studies. qPCR results indicate that both metals together and separately alter cerebral vasculature development at environmental regulatory levels. </p> <p>Lastly, with the increase in the prevalence of neurodegenerative diseases increasing globally, there is a need to evaluate more therapeutics at a high through-put pace and to pinpoint the cause of the sporadic cases. CRISPR-Cas9 technology offers a relatively inexpensive, reliable, and precise advantages over its predecessors when it comes to producing mutant disease models of neurodegeneration. A method to create the expression vector needed for creation of a CRISPR Cas9 knock-in model is detailed in this dissertation. The methodology to insert a chimeric DNA sequence contain human DNA has been created, and the <em>in silico</em> assays used to produce the reactant for this methodology were successful. It has been determined that the efficiency of this knock-in method is limited to the success of producing the chimeric model which is limited itself by the number of molecules included into the chimeric sequence. Overall, the results show that the overlap primers designed in silico need to be re-designed to improve efficiency of the initial reactions required to produce the first plasmid containing the required machinery for a successful knock in of exogenous human DNA.</p>
13

Leveraging the African clawed frog (Xenopus laevis) for Understanding Stage- and Sex-Specific Toxicokinetics and Effects of PFAS

Meredith Norris Scherer (15361759) 26 April 2023 (has links)
<p>Per- and polyfluoroalkyl substances (PFAS) are a group of emerging global contaminants used in a variety of industrial processes and consumer products, such as personal care products and fast-food wrappers. However, due to their carbon-fluorine bonds, these chemicals resist degradation and persist in the environment. PFAS toxicity is driven by a compound’s functional group and chain length with perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHxS), perfluorooctanoic acid (PFOA), and hexafluoropropylene oxide dimer acid (GenX) being of focal concern due to their toxicity to wildlife and presence in the environment. Despite growing concern regarding these contaminants, inadequate attention has been given to evaluating what organismal characteristics influence uptake and depuration of these chemicals, such as life stage and sex. <em>Xenopus laevis</em> tadpoles are a useful model to assess the influence of sex on PFAS kinetics since they have a life history that includes a gill to lung transition. Previous studies have shown that air-breathing organisms depurate PFAS more slowly than water-breathing organisms, but this relationship has never been directly tested. Sex has been shown to be an important factor in the depuration of PFOA for rats, with female rats depurating PFOA in four hours while males depurate in four days. The early portion of bioaccumulation curves are also understudied even though tadpoles accumulate PFAS rapidly, reaching steady state within 48 hours of exposure. <em>Xenopus laevis</em> are used to study multiple endpoints for endocrine disrupting chemicals including PFAS. Despite this, toxicity reference values (TRVs) have not been described for the uptake and elimination of PFAS using <em>X. laevis</em>. To address these gaps in knowledge, I first developed TRVs for <em>X. laevis</em> tadpoles exposed to PFOA throughout metamorphosis and evaluated the influence of sex on phenotypic endpoints. Results showed a no observed effect concentration (NOEC) of 11.1 ppm for body mass at day 14 and no effect of sex on apical endpoints. Next, I described the early bioaccumulation of four PFAS with differing structure (chain lengths and functional groups). PFOS was the only chemical to bioaccumulate with a log bioconcentration factor (BCF) at 10 and 1,000 ppm of 1.33 and 1.18, respectively. PFHxS, PFOA, and GenX had BCFs less than 0. Finally, I examined the impact of life-stage and sex on <em>X. laevis</em> tadpole and juvenile depuration rates. Larval tadpoles depurated four times faster than juveniles, indicating a significant effect of life stage on elimination rates. Sex had no influence on elimination rates. These are the first studies conducted evaluating the significance of life stage and sex in toxicokinetics of PFAS in amphibians.</p>
14

The Plasma Membrane Calcium-ATPase in Mammary Gland Epithelial Cell Lines and Consequences of its Inhibition in a Model of Breast Cancer

Lee, Won Jae Unknown Date (has links)
Ionized calcium (Ca2+), acting as an intracellular messenger, controls numerous biological processes that are essential for life. However, it is also able to convey signals that result in cell death. The fidelity of Ca2+ as a universal second messenger therefore depends on mechanisms that specifically and dynamically regulate its levels within a cell, as well as maintain resting intracellular Ca2+ concentration ([Ca2+]i) very low. One such mechanism for Ca2+ signaling and homeostasis is the plasma membrane Ca2+-ATPase (PMCA), which is a primary active Ca2+ transporter that translocates Ca2+ from a low intracellular Ca2+ environment to a high extracellular environment. There are four mammalian PMCA isoforms (PMCA1−4), which are differentially expressed depending on tissue or cell type. PMCA isoforms possess different sensitivities to biochemical regulation of Ca2+ efflux activity and are also able to subtly alter the dynamics of Ca2+ signals. These properties suggest that the PMCA is not merely a trivial mechanism for Ca2+ extrusion but is influential in contributing to the Ca2+ signaling requirements and unique physiology of different cells. The indispensable nature of Ca2+ signaling in organs such as the brain, heart and skeletal muscle has been the studied extensively but little is known about the roles and regulation of Ca2+ in the mammary gland. This is despite the fact that the mammary gland is a site of extensive Ca2+ flux during lactation. However, cumulating evidence indicates that upregulation of PMCA2 expression in the mammary gland is a major mechanism for milk Ca2+ enrichment. Therefore, the PMCA is likely to be an important mediator of bulk Ca2+ homeostasis in the mammary gland. Studies in other model systems also suggest that PMCAs may regulate other cellular processes such as cell proliferation, differentiation and apoptosis that are required for normal mammary gland physiology. These basic cellular processes are also disturbed in breast cancer and hence deregulation of PMCA expression in the mammary gland may have pathophysiological consequences. Previous studies show that PMCA1 expression is greater in tumorigenic MCF-7 and MDA-MB-231 human breast cancer cells compared to non-tumorigenic MCF-10A human breast epithelial cells. Furthermore, the expression of PMCA1b and PMCA4b is lower in human skin and lung fibroblasts neoplastically transformed by simian virus 40, compared to non-transformed counterparts. It is therefore hypothesized that regulation of PMCA isoform expression is disrupted in breast cancer and that inhibition of PMCA expression in an in vitro model of breast cancer has important effects in modulating intracellular Ca2+ homeostasis, cell proliferation, differentiation and apoptosis. This thesis describes the use of real time RT-PCR to compare PMCA isoform mRNA expression in tumorigenic and non-tumorigenic mammary gland epithelial cells. It demonstrates that particular breast cancer cell lines overexpress PMCA2, an isoform with restricted tissue distribution and which is present in abundant amounts in the lactating rat mammary gland. Thus, some breast cancers may be characterized by the overexpression of Ca2+ transporters that are normally upregulated during the physiological course of lactation. The pathophysiological significance of PMCA2 overexpression in breast cancer is uncertain and future investigations should look at whether levels of PMCA isoform expression correlate with malignancy, prognosis or survival. To address the second hypothesis of this thesis, a stable MCF-7 Tet-off human breast cancer cell line able to conditionally express PMCA antisense was generated. This strategy was necessary due to the current lack of specific pharmacological inhibitors of the PMCA. This thesis shows that PMCA antisense expression significantly inhibits PMCA protein expression, while subtly affecting PMCA-mediated Ca2+ efflux without causing cell death. However, it also reveals that inhibition of PMCA expression has major effects in mediating cell proliferation and cell cycle progression. Moderate changes in PMCA expression and PMCA-mediated Ca2+ transport result in dramatic consequences in MCF-7 cell proliferation. These studies not only support the supposition that modulation of Ca2+ signaling is a viable therapeutic approach for breast cancer but also suggest that PMCAs are possible drug targets. Alternatively, inhibitors of the PMCA may act as adjuvants to augment the efficacy of other anti-neoplastic agents like tamoxifen that have been shown to modulate Ca2+ signaling. Since the discovery of a new family of primary active Ca2+ transporters, which are related to PMCAs, the opportunities in this field of research are very promising.
15

The Plasma Membrane Calcium-ATPase in Mammary Gland Epithelial Cell Lines and Consequences of its Inhibition in a Model of Breast Cancer

Lee, Won Jae Unknown Date (has links)
Ionized calcium (Ca2+), acting as an intracellular messenger, controls numerous biological processes that are essential for life. However, it is also able to convey signals that result in cell death. The fidelity of Ca2+ as a universal second messenger therefore depends on mechanisms that specifically and dynamically regulate its levels within a cell, as well as maintain resting intracellular Ca2+ concentration ([Ca2+]i) very low. One such mechanism for Ca2+ signaling and homeostasis is the plasma membrane Ca2+-ATPase (PMCA), which is a primary active Ca2+ transporter that translocates Ca2+ from a low intracellular Ca2+ environment to a high extracellular environment. There are four mammalian PMCA isoforms (PMCA1−4), which are differentially expressed depending on tissue or cell type. PMCA isoforms possess different sensitivities to biochemical regulation of Ca2+ efflux activity and are also able to subtly alter the dynamics of Ca2+ signals. These properties suggest that the PMCA is not merely a trivial mechanism for Ca2+ extrusion but is influential in contributing to the Ca2+ signaling requirements and unique physiology of different cells. The indispensable nature of Ca2+ signaling in organs such as the brain, heart and skeletal muscle has been the studied extensively but little is known about the roles and regulation of Ca2+ in the mammary gland. This is despite the fact that the mammary gland is a site of extensive Ca2+ flux during lactation. However, cumulating evidence indicates that upregulation of PMCA2 expression in the mammary gland is a major mechanism for milk Ca2+ enrichment. Therefore, the PMCA is likely to be an important mediator of bulk Ca2+ homeostasis in the mammary gland. Studies in other model systems also suggest that PMCAs may regulate other cellular processes such as cell proliferation, differentiation and apoptosis that are required for normal mammary gland physiology. These basic cellular processes are also disturbed in breast cancer and hence deregulation of PMCA expression in the mammary gland may have pathophysiological consequences. Previous studies show that PMCA1 expression is greater in tumorigenic MCF-7 and MDA-MB-231 human breast cancer cells compared to non-tumorigenic MCF-10A human breast epithelial cells. Furthermore, the expression of PMCA1b and PMCA4b is lower in human skin and lung fibroblasts neoplastically transformed by simian virus 40, compared to non-transformed counterparts. It is therefore hypothesized that regulation of PMCA isoform expression is disrupted in breast cancer and that inhibition of PMCA expression in an in vitro model of breast cancer has important effects in modulating intracellular Ca2+ homeostasis, cell proliferation, differentiation and apoptosis. This thesis describes the use of real time RT-PCR to compare PMCA isoform mRNA expression in tumorigenic and non-tumorigenic mammary gland epithelial cells. It demonstrates that particular breast cancer cell lines overexpress PMCA2, an isoform with restricted tissue distribution and which is present in abundant amounts in the lactating rat mammary gland. Thus, some breast cancers may be characterized by the overexpression of Ca2+ transporters that are normally upregulated during the physiological course of lactation. The pathophysiological significance of PMCA2 overexpression in breast cancer is uncertain and future investigations should look at whether levels of PMCA isoform expression correlate with malignancy, prognosis or survival. To address the second hypothesis of this thesis, a stable MCF-7 Tet-off human breast cancer cell line able to conditionally express PMCA antisense was generated. This strategy was necessary due to the current lack of specific pharmacological inhibitors of the PMCA. This thesis shows that PMCA antisense expression significantly inhibits PMCA protein expression, while subtly affecting PMCA-mediated Ca2+ efflux without causing cell death. However, it also reveals that inhibition of PMCA expression has major effects in mediating cell proliferation and cell cycle progression. Moderate changes in PMCA expression and PMCA-mediated Ca2+ transport result in dramatic consequences in MCF-7 cell proliferation. These studies not only support the supposition that modulation of Ca2+ signaling is a viable therapeutic approach for breast cancer but also suggest that PMCAs are possible drug targets. Alternatively, inhibitors of the PMCA may act as adjuvants to augment the efficacy of other anti-neoplastic agents like tamoxifen that have been shown to modulate Ca2+ signaling. Since the discovery of a new family of primary active Ca2+ transporters, which are related to PMCAs, the opportunities in this field of research are very promising.
16

Identification of Neurotoxic Targets of Diverse Chemical Classes of Dietary Neurotoxins/Neurotoxicants

Rachel M Foguth (9343949) 16 December 2020 (has links)
<p>Neurological disorders are a major public health concern due to prevalence, severity of symptoms, and impact on caregivers and economic losses. While genetic susceptibility likely has a role in most cases, exposure to toxicants can lead to neurotoxicity, including potentially developmental origins of adult disease or increased risk of disease onset. These exposures are not necessarily large, acute exposures, but could accumulate, with a chronic low-dose exposure, causing toxicity. This research focuses on the potential neurotoxicity of two classes of dietary toxins/toxicants, heterocyclic aromatic amines (HAAs) and per- and polyfluoroalkyl substances (PFAS). HAAs, such as PhIP, harmane, and harmine, are formed in charred or overcooked meat, coffee, tobacco, and other foods. PFAS are largely used in making household materials, but are found in small amounts in eggs and dairy products and largely in contaminated water. While these two classes are diverse in terms of structure, common neurotoxic targets and mechanisms often exist. Therefore, we tested the effects of these chemicals on cell viability and neurotoxicity. In the first aim, we aimed to elucidate the mechanism of toxicity of harmane and harmine, focusing on their ability to cause mitochondrial dysfunction. The second aim was to determine the effects of either harmane or PhIP on the nigrostriatal motor systems and motor function of rats and mice, respectively. The third aim determined the effects of PFAS on neurodevelopment of Northern leopard frogs, focusing on changes in neurotransmitter levels and accumulation in the brain. Harmane did not cause motor dysfunction, but potentially affected the nigro-striatal motor system in an age- or sex-dependent manner. PhIP had differential effects on dopamine levels over time and caused motor dysfunction after subchronic exposure in mice. Perfluorooctane sulfonate (PFOS) accumulated in the brains of frogs and PFAS caused changes in neurotransmitter levels that were dose- and time-dependent. Overall, this research shows that toxins/toxicants humans are exposed to over their whole lives through their diet and contaminated water can cause neurotoxicity, potentially leading to or increasing risk of disease states. </p>
17

Developmental Toxicity Assessment of Perfluoroalkyl Substances (PFAS) Using Zebrafish Model System

Ola Wasel (13158639) 27 July 2022 (has links)
<p>  </p> <p>Perfluoroalkyl substances (PFAS) are synthetic chemicals that are composed of fluorinated aliphatic chains and are widely used in industrial and consumer products. These chemicals are very stable and persist in the environment. Due to concerns linked with longer chain PFAS, shorter chain chemicals are being used as replacements. There are limited human health data regarding the shorter chain chemicals. In addition, these alternatives are persistent in the environment similar to the longer chain PFAS. The main objective of this dissertation was to assess developmental toxicity of the shorter chain PFAS or shorter chain PFAS with chemical modifications represented by perfluorobutanoic acid (PFBA, C4), perfluorohexanoic acid (PFHxA, C6), perfluorobutane sulfonate (PFBS, C4), and perfluoro-2-proxypropanoic acid (GenX, C6). Overall, the results showed that chain length and functional group are determinants of toxicity of PFAS. All tested PFAS induced one or more developmental adverse outcome, but the effects of each chemical are unique, warranting further studies to address the toxicity of the replacement PFAS. </p>
18

<b>Heavy Metal Concentrations in Sea Turtles and </b><b>Their Prey in the Northwest Atlantic </b>

Yi Wynn Chan (18414897) 20 April 2024 (has links)
<p dir="ltr">The Northwest Atlantic Ocean, which surrounds the US eastern coastline, is an area rich in marine life. The US eastern coastline is also highly urbanized, resulting in a lot of pollutants (like heavy metals) entering the marine environment. This is of concern for long-lived marine species like sea turtles. Since sea turtles are long-lived and highly migratory, their tissues can often incorporate these pollutants through environmental and dietary exposure. I collected tissue samples from 5 different sea turtle populations in the Northwest Atlantic and analyzed them for concentrations of silver (Ag), aluminum (Al), arsenic (As), cadmium (Cd), cobalt (Co), chromium (Cr), iron (Fe), manganese (Mn), nickel (Ni), lead (Pb), selenium (Se) and zinc (Zn) using an Inductively Coupled Plasma Mass Spectrometry (ICP-MS). The first chapter looks at skin (reflects exposure ~1 year ago) and scute (reflects exposure from 4-6 years ago) samples collected during necropsies of juvenile green (<i>Chelonia mydas</i>) (n=8), Kemp’s ridley (<i>Lepidochelys kempii</i>) (n=30) and loggerhead (<i>Caretta caretta</i>) (n=17) turtles that were found cold-stunned in Cape Cod Bay, Massachusetts. In scute samples, the heavy metal with the highest concentration for green turtles was iron, zinc for loggerhead turtles, and arsenic for Kemp’s ridley turtles. In skin samples, the heavy metal with the highest concentration for green turtles was iron, arsenic for loggerhead turtles, and aluminum for Kemp’s ridley turtles. Overall, I found scute samples to have higher heavy metal concentrations than skin samples. The second chapter looks at scute samples collected from loggerhead turtles of different life stages. These samples were collected during necropsies of cold-stunned loggerhead turtles from Cape Cod Bay, Massachusetts (CCB; n=17), as well as from live loggerhead turtles in the Mid-Atlantic Bight (MAB; n=37) and off the coast of North Carolina (NC; n=9). We also collected commonly known loggerhead turtle prey items including whelk (<i>Buccinum undatum</i>) (n=12), Atlantic scallop (<i>Placopecten magellanicus)</i> (n=10) and Jonah crab (<i>Cancer borealis</i>) (n=5) from the Mid-Atlantic Bight region to study the occurrence of biomagnification through trophic pathways. NC loggerhead turtles had higher heavy metal concentrations than other locations except for cadmium and zinc, where CCB loggerhead turtles were higher. I found that all heavy metals except silver, cadmium, and lead appear to be biomagnified (TTF>1) in loggerhead turtles. These two chapters provided baseline information on heavy metal concentrations in sea turtles in east coast US.</p>
19

The Plasma Membrane Calcium-ATPase in Mammary Gland Epithelial Cell Lines and Consequences of its Inhibition in a Model of Breast Cancer

Lee, Won Jae Unknown Date (has links)
Ionized calcium (Ca2+), acting as an intracellular messenger, controls numerous biological processes that are essential for life. However, it is also able to convey signals that result in cell death. The fidelity of Ca2+ as a universal second messenger therefore depends on mechanisms that specifically and dynamically regulate its levels within a cell, as well as maintain resting intracellular Ca2+ concentration ([Ca2+]i) very low. One such mechanism for Ca2+ signaling and homeostasis is the plasma membrane Ca2+-ATPase (PMCA), which is a primary active Ca2+ transporter that translocates Ca2+ from a low intracellular Ca2+ environment to a high extracellular environment. There are four mammalian PMCA isoforms (PMCA1-4), which are differentially expressed depending on tissue or cell type. PMCA isoforms possess different sensitivities to biochemical regulation of Ca2+ efflux activity and are also able to subtly alter the dynamics of Ca2+ signals. These properties suggest that the PMCA is not merely a trivial mechanism for Ca2+ extrusion but is influential in contributing to the Ca2+ signaling requirements and unique physiology of different cells. The indispensable nature of Ca2+ signaling in organs such as the brain, heart and skeletal muscle has been the studied extensively but little is known about the roles and regulation of Ca2+ in the mammary gland. This is despite the fact that the mammary gland is a site of extensive Ca2+ flux during lactation. However, cumulating evidence indicates that upregulation of PMCA2 expression in the mammary gland is a major mechanism for milk Ca2+ enrichment. Therefore, the PMCA is likely to be an important mediator of bulk Ca2+ homeostasis in the mammary gland. Studies in other model systems also suggest that PMCAs may regulate other cellular processes such as cell proliferation, differentiation and apoptosis that are required for normal mammary gland physiology. These basic cellular processes are also disturbed in breast cancer and hence deregulation of PMCA expression in the mammary gland may have pathophysiological consequences. Previous studies show that PMCA1 expression is greater in tumorigenic MCF-7 and MDA-MB-231 human breast cancer cells compared to non-tumorigenic MCF-10A human breast epithelial cells. Furthermore, the expression of PMCA1b and PMCA4b is lower in human skin and lung fibroblasts neoplastically transformed by simian virus 40, compared to non-transformed counterparts. It is therefore hypothesized that regulation of PMCA isoform expression is disrupted in breast cancer and that inhibition of PMCA expression in an in vitro model of breast cancer has important effects in modulating intracellular Ca2+ homeostasis, cell proliferation, differentiation and apoptosis. This thesis describes the use of real time RT-PCR to compare PMCA isoform mRNA expression in tumorigenic and non-tumorigenic mammary gland epithelial cells. It demonstrates that particular breast cancer cell lines overexpress PMCA2, an isoform with restricted tissue distribution and which is present in abundant amounts in the lactating rat mammary gland. Thus, some breast cancers may be characterized by the overexpression of Ca2+ transporters that are normally upregulated during the physiological course of lactation. The pathophysiological significance of PMCA2 overexpression in breast cancer is uncertain and future investigations should look at whether levels of PMCA isoform expression correlate with malignancy, prognosis or survival. To address the second hypothesis of this thesis, a stable MCF-7 Tet-off human breast cancer cell line able to conditionally express PMCA antisense was generated. This strategy was necessary due to the current lack of specific pharmacological inhibitors of the PMCA. This thesis shows that PMCA antisense expression significantly inhibits PMCA protein expression, while subtly affecting PMCA-mediated Ca2+ efflux without causing cell death. However, it also reveals that inhibition of PMCA expression has major effects in mediating cell proliferation and cell cycle progression. Moderate changes in PMCA expression and PMCA-mediated Ca2+ transport result in dramatic consequences in MCF-7 cell proliferation. These studies not only support the supposition that modulation of Ca2+ signaling is a viable therapeutic approach for breast cancer but also suggest that PMCAs are possible drug targets. Alternatively, inhibitors of the PMCA may act as adjuvants to augment the efficacy of other anti-neoplastic agents like tamoxifen that have been shown to modulate Ca2+ signaling. Since the discovery of a new family of primary active Ca2+ transporters, which are related to PMCAs, the opportunities in this field of research are very promising.
20

Nutritional and toxicological studies on New Zealand mutton bird meat (Puffinus griseus)

Al-Amer, Saleh Suliaman January 2009 (has links)
New Zealand mutton bird or tītī (Puffinus griseus and order procellariiformes) nest in New Zealand during the summer months, migrate to the northern hemisphere during May and return in September. Their eggs are laid during November and December and the chicks are hatched in the following January and February. Large numbers of them are harvested from April to May in New Zealand. They are wild seabirds annually harvested by Maori according to the customary rights agreement set by Treaty of Waitangi.NZ mutton birds also called Sooty Shearwaters are noted for their high proportion of body fat.These birds are interesting since its sole diet is based on krill and other small marine organisms that are potentially rich in n-3 fatty acids and other marine bioactive compounds. The proximate composition, fatty and amino acids and cholesterol content of mutton bird pectoral muscle were determined and compared with other common meat to explore the nutritional value of this New Zealand delicacy. The concentration of twenty two essential and toxic elements including silver (Ag), aluminium (Al), arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), manganese (Mn), lead (Pb), selenium (Se), potassium (K), calcium (Ca), magnesium (Mg), boron (B), iron (Fe), nickel (Ni), sulphur (S), sodium (Na) and cobalt (Co) and zinc (Zn) in mutton bird breast meat (Puffinus griseus) were measured over two years to evaluate its safety for human consumption. Persistent organochlorine, dichlorodiphenyltrichloroethane (DDT) and their metabolites, and aldrin and lindane were also measured.Twenty bird carcasses were purchased in both 2006 and 2007 from a local source. Meat samples from the pectoral muscle of two carcasses were pooled to generate 10 samples for each year. These were used for trace element analysis using inductively coupled plasma-mass spectrometry (ICP-MS). Trace elements were in the range of 0 to 1.09 mg/kg wet weight for Ag, 0 to 3.32 for Al, 0.17 to 0.79 for As, 0.01 to 0.07 for Cd, 0.03 to 0.15 for Cr, 3.56 to 4.88 for Cu, 0 to 0.15 for Hg, 0.22 to 0.50 for Mn, 0 to 0.09 for Pb, 0.66 to 1.18 for Se and 11.49 to 23.70 for Zn. In 2006, Ag, Al, Mn and Zn concentrations were significantly higher but Pb and Hg concentrations significantly lower compared to the 2007 samples (P < 0.05). Apart from one sample in 2006, all the samples were below the published maximum level for concern. However, our preliminary data indicated that the higher level of Cd and other metals in the skin of mutton bird may compromise the overall safety to humans consuming the skin of mutton birds. It is suggested that the evaluation of the metals in different parts and/or the whole mutton bird at different seasons is required to assure complete safety to the consumers.Furthermore, the nutritional value of mutton bird meat was studied over two harvesting seasons (2006 and 2007) to investigate the impact of seasonal variation. The moisture and carbohydrates contents ranged between 54.0 to 55.0 % and 2.8 to 3.0 %, respectively, and no seasonal effects were evident in these components. The values for fat and ash contents were higher and the protein content lower for birds harvested in 2007 compared with the 2006 values which ranged from 11.8 to 13.0, 10.3 to 11.7, and 20.3 to 18.5 % for fat, ash and protein content respectively. The major amino acids in mutton bird pectoral muscle were glutamate, aspartate, lysine, leucine, and arginine. Higher lysine concentrations and lower proline, cystein and methionine were found in mutton birds compared with the literature values for beef, lamb and pork. The essential amino acid content in mutton bird (43.8 and 44.9 % in 2006 and 2007, respectively) was slightly higher than those found in beef and lamb meats (42-43%).The major fatty acids detected were palmitic (C16:0), stearic (C18:0), oleic and isomers (C18:1), eicosenoic (C20:1), Docosahexaenoic acid (DHA) (C22:6), icosapentaenoic acid (EPA) (C 20:5) and these accounted for approximately 77% of the fatty acids. The 3/6 ratio of fats from pectoral muscle was 1.3. The cholesterol concentration varied slightly in the two years with 184.4±37.37and 134.4±25.55mg/100 g fresh weight for 2007 and 2008 respectively. Mutton bird was shown to contain significantly higher cholesterol content (134.4-184.4) than other common meat such as chicken (80.3-88.9), lamb (62.3), fish (52.79) and beef (51.97). Overall, the nutritional value of mutton bird muscle was similar to or superior to the traditionally protein sources such as seafood and red meat. Annual variations existed in the composition of Mutton bird pectoral muscle but this is not of nutritional consequence but might be a useful indicator for ecological events such as feed availability and other environmental issues. Mutton bird seems to be a good source of essential minerals, Zn and Fe compared with other traditional meats source. Mutton bird meat is nutritionally as good as the major sources of red or white meats. It may even have advantages over the other common meats (beef, lamb, fish and chicken) due to its high protein and monounsaturated fatty acids (omega n-3 and n-6) content. However, its high cholesterol content may represent a risk factor for some people.

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