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 Department: Department of Biological Sciences
N-Acylethanolamine metabolism during seed germination: Molecular identification of a functional N-acylethanolamine amidohydrolase.

N-Acylethanolamine metabolism during seed germination: Molecular identification of a functional N-acylethanolamine amidohydrolase.

Date: August 2004
Creator: Shrestha, Rhidaya
Description: N-Acylethanolamines (NAEs) are endogenous lipid metabolites that occur in a variety of dry seeds, and their levels decline rapidly during the first few hours of imbibition (Chapman et al., 1999, Plant Physiol., 120:1157-1164). Biochemical studies supported the existence of an NAE amidohydrolase activity in seeds and seedlings, and efforts were directed toward identification of DNA sequences encoding this enzyme. Mammalian tissues metabolize NAEs via an amidase enzyme designated fatty acid amide hydrolase (FAAH). Based on the characteristic amidase signature sequence in mammalian FAAH, a candidate Arabidopsis cDNA was identified and isolated by reverse transcriptase-PCR. The Arabidopsis cDNA was expressed in E. coli and the recombinant protein indeed hydrolyzed a range of NAEs to free fatty acids and ethanolamine. Kinetic parameters for the recombinant protein were consistent with those properties of the rat FAAH, supporting identification of this Arabidopsis cDNA as a FAAH homologue. Two T-DNA insertional mutant lines with disruptions in the Arabidopsis NAE amidohydrolase gene (At5g64440) were identified. The homozygous mutant seedlings were more sensitive than the wild type to exogenously applied NAE 12:0. Transgenic seedlings overexpressing the NAE amidohydrolase enzyme showed noticeably greater tolerance to NAE 12:0 than wild type seedlings. These results together provide evidence in vitro ...
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N-Acylethanolamine (NAE) profiles change during Arabidopsis thaliana seed germination and seedling growth.

N-Acylethanolamine (NAE) profiles change during Arabidopsis thaliana seed germination and seedling growth.

Date: August 2006
Creator: Wiant, William C.
Description: An understanding of the potential roles as lipid mediators of a family of bioactive metabolites called N-acylethanolamines (NAEs) depends on their accurate identification and quantification. The levels of 18C unsaturated NAEs (e.g. NAE18:2, NAE 18:3, etc.) in wild-type seeds (about 2000 ng/g fw) generally decreased by about 80% during germination and post-germinative growth. In addition, results suggest NAE-degradative fatty acid amide hydrolase (FAAH) expression does not play a major role in normal NAE metabolism as previously thought. Seedlings germinated and grown in the presence of abscisic acid (ABA), an endogenous plant hormone, exhibited growth arrest and secondary dormancy, similar to the treatment of seedlings with exogenous N­lauroylethanolamine (NAE12:0). ABA-mediated growth arrest was associated with higher levels of unsaturated NAEs. Overall, these results are consistent with the concept that NAE metabolism is activated during seed germination and suggest that the reduction in unsaturated NAE levels is under strict temporal control and may be a requirement for normal seed germination and post-germinative growth.
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Adaptive Advantages of Carotenoid Pigments in Alpine and Subalpine Copepod Responses to Polycyclic Aromatic Hydrocarbon Induced Phototoxicity

Adaptive Advantages of Carotenoid Pigments in Alpine and Subalpine Copepod Responses to Polycyclic Aromatic Hydrocarbon Induced Phototoxicity

Date: May 2010
Creator: Kovach, Matthew James
Description: Alpine zooplankton are exposed to a variety of stressors in their natural environment including ultraviolet radiation. Physiological coping mechanisms such as the accumulation of photoprotective compounds provide these zooplankton protection from many of these stressors. Elevated levels of carotenoid compounds such as astaxanthin have been shown to help zooplankton survive longer when exposed to ultraviolet radiation presumably due to the strong antioxidant properties of carotenoid compounds. This antioxidant capacity is important because it may ameliorate natural and anthropogenic stressor-induced oxidative stress. While previous researchers have shown carotenoid compounds impart increased resistance to ultraviolet radiation in populations of zooplankton, little work has focused on the toxicological implications of PAH induced phototoxicity on zooplankton containing high levels of carotenoid compounds. This thesis discusses research studying the role that carotenoid compounds play in reducing PAH induced phototoxicity. By sampling different lakes at elevations ranging from 9,500' to 12,700' in the front range of the Colorado Rocky Mountains, copepod populations containing different levels of carotenoid compounds were obtained. These populations were then challenged with fluoranthene and ultraviolet radiation. Results discussed include differences in survival and levels of lipid peroxidation among populations exhibiting different levels of carotenoid compounds, and the toxicological and ecological implications of ...
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The adolescent stress response to a naturalistic driving stressor

The adolescent stress response to a naturalistic driving stressor

Date: August 2000
Creator: Wingo, Mary
Description: The proposed study examined the role of anxiety and risk-taking in driving performance in adolescents. In addition to examining the sample as a whole, gender differences were assessed given earlier reports from our laboratory and others indicating that males and females differ with respect to risky behaviors to driving performance and anxiety. Adolescents' subjective and physiological responses to a driving simulator task were assessed. Anxiety was measured via self report and salivary cortisol. Participants provided a baseline saliva sample and 3 post-task samples for cortisol analysis. Subjective anxiety scores were obtained at both baseline and following the driving stressor. Information concerning impulsivity, as well as other psychological constructs was also collected at baseline. Unlike the pilot study, there were no relationships (with or without respect to gender) between salivary cortisol and both self-reported anxiety (state and trait) or impulsively measures for this sample. These results suggest that this group of adolescents may not have been anxious about the driving task. This discrepancy may stem from error introduced by the smaller sample size obtained from the initial findings or to other factors remaining outside the parameters of the current study. The task did, however, induce a slight hypothalamic pituitary adrenal (HPA) ...
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Age-Dependent Effects Of Chronic GABAA  Receptor Blockade In Barrel Cortex

Age-Dependent Effects Of Chronic GABAA Receptor Blockade In Barrel Cortex

Date: May 2001
Creator: Gargan, Lynn
Description: GABAA receptor binding is transiently increased in rat whisker barrels during the second postnatal week, at a time when neurons in the developing rat cortex are vulnerable to excitotoxic effects. To test whether these GABAA receptors might serve to protect neurons from excessive excitatory input, polymer implants containing the GABAA receptor antagonist bicuculline were placed over barrel cortex for a 4-day period in young (postnatal days 8 - 12) and adult rats. In the cortex of young, but not adult rats, the chronic blockade of GABAA receptors resulted in substantial tissue loss and neuron loss. The greater loss of neurons in young rats supports the hypothesis that a high density of GABAA receptors protects neurons from excessive excitatory input during a sensitive period in development.
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Aging Is a Determinant in Anoxia Stress Tolerance in Caenorhabditis Elegans

Aging Is a Determinant in Anoxia Stress Tolerance in Caenorhabditis Elegans

Date: May 2013
Creator: Goy, Jo M.
Description: Oxygen availability is critical for survival for most organisms. The nematode, C. elegans, has been useful for studying genetic regulation of anoxia tolerance due to the oxygen deprivation response mechanisms shared with other metazoans. Studies examining long-term anoxia (72h, LTA) tolerance have only been conducted at adult day 1. To investigate the effect of aging on anoxia tolerance wild-type and mutant strains were exposed to LTA between adult day 1 and day 9. Wild-type isolates and daf-16(mu86) (FOXO transcription factor regulated by insulin-signaling) and aak-2(gt33) (catalytic subunit of AMP-activated protein kinase) strains were anoxia sensitive at day 1 and displayed increased LTA tolerance with aging correlated with reproductive senescence followed by a decline in survivorhsip through day 9. The daf-2(e1370) (insulin receptor homologue of C. elegans), glp-1(e2141) (a lin-12/Notch receptor) and fog-2(q71) (required for spermatogenesis) strains were LTA-tolerant through day 5. I conclude that aging influences LTA-tolerance in a strain- and age-dependent manner. In addition to being LTA-tolerant the daf-2(e1370) and glp-1(e2141) strains have a longevity phenotype that is suppressed by loss of kri-1 or daf-12. While loss of kri-1 did not suppress the LTA-tolerant phenotype of glp-1(e2141) at day 1 the portion of impaired survivors increased at day 3 ...
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Alterations in Fatty Acid Amide Hydrolase (Faah) Transcript Levels and Activity Lead to Changes in the Abiotic Stress Susceptibility of Arabidopsis Thaliana

Alterations in Fatty Acid Amide Hydrolase (Faah) Transcript Levels and Activity Lead to Changes in the Abiotic Stress Susceptibility of Arabidopsis Thaliana

Date: May 2012
Creator: Gonzalez, Gabriel
Description: N-Acylethanolamines (NAEs) are a class of bioactive lipids, and FAAH is one of the enzymes responsible for degrading NAEs in both plants and animals. in plants, FAAH appears to be closely associated with ABA, a phytohormone which has long been associated with plant stress responses, since the overexpression of FAAH in Arabidopsis results in ABA hypersensitivity. Therefore, it is reasonable to speculate that alterations in FAAH transcript levels will result in altered stress responses in plants. to investigate this hypothesis experiments were carried out in which wild type (WT), FAAH-overexpressing (OE), and T-DNA insertional FAAH knockouts of Arabidopsis (faah) were grown in MS media under stress conditions. the stress conditions tested included chilling stress, heavy metal stress induced by cadmium or copper, nutrient limitations induced by low phosphorus or low nitrogen, salt stress induced with NaCl, and osmotic stress induced with mannitol. the OE plants were consistently hypersensitive to all stress conditions in relation to wild type plants. Inactive FAAH overexpressors did not have the hypersensitivity to the salt and osmotic stress of the active OE plants and were instead tolerant to these stresses. FAAH2 (faah2) knockouts and FAAH 1 and 2 double knockouts (faah 1+2) were based on some ...
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Analysis and Development of Post Secondary Curriculum on Sustainability

Analysis and Development of Post Secondary Curriculum on Sustainability

Date: May 2000
Creator: White, Miki Machell
Description: This thesis examines existing curricula at colleges and universities about sustainability and uses results to develop an introductory post secondary course curriculum. The proposed course is organized around three major elements - - science, philosophy, and economics - - all integral to understanding sustainability. Materials needed to teach the proposed 3-semester hour course including syllabus, teaching modules, transparencies, handouts, and exams were developed. Suggestions on how to teach a one-semester hour course on sustainability and a workshop on sustainability are also presented. The following research and curriculum development was a project established and funded by the Texas Energy Office, Renewable Resources and Sustainability Program.
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Analysis and expression of the cotton gene for the D-12 fatty acid desaturases 2-4 (FAD2-4)

Analysis and expression of the cotton gene for the D-12 fatty acid desaturases 2-4 (FAD2-4)

Date: August 2003
Creator: Park, Stacy J.
Description: A genomic clone containing a 16.9-kb segment of cotton DNA was found to encompass a D-12 fatty acid desaturases (FAD2-4) gene. The FAD2-4 gene has a single, large intron of 2,780 bp in its 5'-untranslated region, just 12 bp upstream from the ATG initiation codon of the FAD2-4 opening reading frame. A number of prospective promoter elements, including several light-responsive sequences, occur in the 5'-flanking region. The coding region of the gene is 1155 bp with no introns, and would encode a FAD2-4 polypeptide of 384 amino acids. The putative protein had four membrane-spanning helices, hallmarks of an integral membrane protein, and would probably be located in the endoplasmic reticulum. The FAD2-4 gene is indeed a functional gene, since yeast cells transformed with a plasmid containing the coding region of the gene synthesize an appreciable amount of linoleic acid (18:2), not normally made in wild-type yeast cells. The FAD2-4 gene has many structural similarities to the cotton FAD2-3 gene that was also analyzed in this laboratory.
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Analysis of a Cotton Gene Cluster for the Antifungal Protein Osmotin

Analysis of a Cotton Gene Cluster for the Antifungal Protein Osmotin

Date: December 2003
Creator: Wilkinson, Jeffery Roland
Description: Three overlapping genomic clones covering 29.0 kilobases of cotton DNA were found to encompass a cluster of two presumptive osmotin genes (OSMI and OSMII) and two osmotin pseudogenes (OSMIII and OSMIV). A segment of 16,007 basepairs of genomic DNA was sequenced from the overlapping genomic clones (GenBank Accessions AY303690 and AF304007). The two cotton osmotin genes were found to have open reading frames of 729 basepairs without any introns, and would encode presumptive osmotin preproteins of 242 amino acids. The open reading frames of the genes are identical in sequence to two corresponding cDNA clones (GenBank Accessions AF192271 and AY301283). The two cDNA inserts are almost full-length, since one lacks codons for the four N-terminal amino acids, and the other cDNA insert lacks the coding region for the 34 N-terminal amino acids. The cotton osmotin preproteins can be identified as PR5 proteins from their similarities to the deduced amino acid sequences of other plant osmotin PR5 preproteins. The preproteins would have N-terminal signal sequences of 24 amino acids, and the mature 24 kilodalton isoforms would likely be targeted for extracellular secretion. Prospective promoter elements, including two ethylene response elements, implicated as being positive regulatory elements in the expression of a ...
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