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 ...

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 ...

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 ...

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.

Description: This study analyzes the remaining suitable habitat of the one-horned rhinoceros, Rhinoceros unicornis, in Royal Chitwan National Park of Nepal. An April 2003 Landsat image was classified into eight land cover types: wetland, sand, water, mixed forest, sal forest, agriculture, settlement, and grassland. This image was converted into habitat suitability maps using cover, food, and water. The rhinoceros prefers grassland habitat with oxbow lakes and closed canopy during the monsoon season. Nominal values of five parameters were used to create a map of habitat suitability index. The map was categorized into four habitat classes: highly unsuitable, unsuitable, moderately suitable habitat, and suitable. Landscape metrics, patch metrics and class metrics associated with habitat were determined through the use of FRAGSTATS.