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Integrating Selective Herbicide and Native Plant Restoration to Control Alternanthera philoxeroides (Alligator Weed)

Integrating Selective Herbicide and Native Plant Restoration to Control Alternanthera philoxeroides (Alligator Weed)

Date: December 2011
Creator: Adams, Justin
Description: Exotic invasive aquatic weeds such as alligator weed (Alternanthera philoxeroides) threaten native ecosystems by interfering with native plant communities, disrupting hydrology, and diminishing water quality. Development of new tools to combat invaders is important for the well being of these sensitive areas. Integrated pest management offers managers an approach that combines multiple control methods for better control than any one method used exclusively. In a greenhouse and field study, we tested the effects of selective herbicide application frequency, native competitor plant introduction, and their integration on alligator weed. In the greenhouse study, alligator weed shoot, root, and total biomass were reduced with one herbicide application, and further reduced with two. Alligator weed cumulative stem length and shoot/root ratio was only reduced after two applications. In the greenhouse, introduction of competitors did not affect alligator weed biomass, but did affect shoot/root ratio. The interaction of competitor introduction and herbicide did not significantly influence alligator weed growth in the greenhouse study. In the field, alligator weed cover was reduced after one herbicide application, but not significantly more after a second. Introduction of competitor species had no effect on alligator weed cover, nor did the interaction of competitor species and herbicide application. This ...
Contributing Partner: UNT Libraries
The Effects of Glyphosate Based Herbicides on Chick Embryo Development

The Effects of Glyphosate Based Herbicides on Chick Embryo Development

Date: August 2013
Creator: Winnick, Blake Edward
Description: Glyphosate based herbicides are among the most widely used herbicides in the world. The purpose of this study was to determine developmental toxicity of glyphosate, the active ingredient in the common herbicide Roundup, on developing chicken embryos. Few studies have examined toxic effects of glyphosate alone versus the full compound formulations of Roundup, which include adjuvants and surfactants. Adjutants and surfactants are added to aid in solubility and absorption of glyphosate. In this study chicken embryos were exposed at the air cell on embryonic day 6 to 19.8 or 9.9 mg / Kg egg mass of glyphosate in Roundup or glyphosate only. Chickens treated with 19.8 and 9.9 mg / Kg glyphosate in Roundup showed significant reduction in survivability compared to glyphosate alone treatments and controls. On embryonic day 18, embryos were sacrificed for evaluation of developmental toxicity using wet embryo mass, dry embryo mass, and yolk mass as indicators. Morphology measurements were taken on liver mass, heart mass, tibiotarsus length and beak length. Embryos treated with 19.8 mg / Kg glyphosate and 9.9 mg / Kg glyphosate in Roundup showed significant reductions in wet and dry embryo mass and yolk mass. Tibiotarsus length in 9.9 mg / Kg glyphosate ...
Contributing Partner: UNT Libraries
Photoinduced Toxicity in Early Lifestage Fiddler Crab (Uca Longisignalis) Following Exposure to Deepwater Horizon Spill Oil

Photoinduced Toxicity in Early Lifestage Fiddler Crab (Uca Longisignalis) Following Exposure to Deepwater Horizon Spill Oil

Access: Use of this item is restricted to the UNT Community.
Date: December 2015
Creator: Taylor, Leigh M.
Description: The 2010 Deepwater Horizon (DWH) oil spill resulted in a large release of polycyclic aromatic hydrocarbons (PAH) into the Gulf of Mexico. PAH can interact with ultraviolet radiation (UV) resulting in increased toxicity, particularly to early lifestage organisms. The goal of this research was to determine the sensitivity of fiddler crab larvae (Uca longisignalis) to photo-induced toxicity following exposure to Deepwater Horizon spill oil in support of the DWH Natural Resource Damage Assessment. Five replicate dishes each containing 20 larvae, were exposed to one of three UV treatments (10%, 50%, and 100% ambient natural sunlight) and one of five dilutions of water accommodated fractions of two naturally weathered source oils. A dose dependent effect of PAH and UV on larval mortality was observed. Mortality was markedly higher in PAH treatments that included co-exposure to more intense UV light. PAH treatments under low intensity sunlight had relatively high survival. These data demonstrate the importance of considering combined effects of non-chemical (i.e. UV exposure) and chemical stressors and the potential for photo-induced effects after exposure to PAH following the Deepwater Horizon spill.
Contributing Partner: UNT Libraries
Phototoxic Effects of Titanium Dioxide Nanoparticles on Daphnia Magna

Phototoxic Effects of Titanium Dioxide Nanoparticles on Daphnia Magna

Date: December 2013
Creator: Mansfield, Charles M.
Description: Titanium dioxide nanoparticles (TiO2-NP) are one of the most abundantly utilized nanomaterials in the world. Studies have demonstrated the mechanism of acute toxicity in TiO2-NP to be the production of reactive oxygen species (ROS) leading to oxidative stress and mortality in exposed organisms. It has also been demonstrated that the anatase crystalline conformation is capable of catalyzing the cleavage of water molecules to further increase the concentration of ROS in the presence of ultraviolet radiation. This photoenhanced toxicity significantly lowers the toxicity threshold of TiO2-NP to environmentally relevant concentrations (ppb). The goal of this study was to determine whether dietary uptake and accumulation of TiO2-NP in the aquatic filter feeder Daphnia magna resulted in photoenhanced toxicity. D. magna and S. caprincornatum were exposed to aqueous solutions of 20ppm and 200ppm TiO2-NP for 24hrs and then transferred to clean moderately hard water. Samples were taken at various time points, dried, and TiO2 quantified using ICP-MS. Toxicity assays were run on D. magna using three TiO2-NP (20ppm, 200ppm) exposure protocols and two ultraviolet radiation treatments. The first exposure group was exposed to aqueous solutions of TiO2-NP for the duration of the test. The second exposure group was exposed to TiO2-NP for an ...
Contributing Partner: UNT Libraries
Changes in Gene Expression Levels of the Ecf Sigma Factor Bov1605 Under Ph Shift and Oxidative Stress in the Sheep Pathogen Brucella Ovis

Changes in Gene Expression Levels of the Ecf Sigma Factor Bov1605 Under Ph Shift and Oxidative Stress in the Sheep Pathogen Brucella Ovis

Date: December 2012
Creator: Kiehler, Brittany Elaine
Description: Brucella ovis is a sexually transmitted, facultatively anaerobic, intracellular bacterial pathogen of sheep (Ovis aries) and red deer (Cervus elaphus). Brucella spp. infect primarily by penetrating the mucosa and are phagocytized by host macrophages, where survival and replication occurs. At least in some species, it has been shown that entry into stationary phase is necessary for successful infection. Brucella, like other alphaproteobacteria, lack the canonical stationary phase sigma factor ?s. Research on diverse members of this large phylogenetic group indicate the widespread presence of a conserved four-gene set including an alternative ECF sigma factor, an anti-sigma factor, a response regulator (RR), and a histidine kinase (HK). The first description of the system was made in Methylobacterium extorquens where the RR, named PhyR, was found to regulate the sigma factor activity by sequestering the anti-sigma factor in a process termed "sigma factor mimicry." These systems have been associated with various types of extracellular stress responses in a number of environmental bacteria. I hypothesized that homologous genetic sequences (Bov_1604-1607), which are similarly found among all Brucella species, may regulate survival functions during pathogenesis. To further explore the involvement of this system to conditions analogous to those occurring during infection, pure cultures of ...
Contributing Partner: UNT Libraries
Hepatotoxicity of Mercury to Fish

Hepatotoxicity of Mercury to Fish

Date: August 2010
Creator: Barst, Benjamin Daniel
Description: Tissue samples from spotted gar (Lepisosteus oculatus) and largemouth bass (Micropterus salmoides) were collected from Caddo Lake. Gar and bass livers were subjected to histological investigation and color analysis. Liver color (as abs at 400 nm) was significantly correlated with total mercury in the liver (r2 = 0.57, p = 0.02) and muscle (r2 = 0.58, p = 0.01) of gar. Evidence of liver damage as lipofuscin and discoloration was found in both species but only correlated with liver mercury concentration in spotted gar. Inorganic mercury was the predominant form in gar livers. In order to determine the role of mercury speciation in fish liver damage, a laboratory feeding study was employed. Zebrafish (Danio rerio) were fed either a control (0.12 ± 0.002 µg Hg.g-1 dry wt), inorganic mercury (5.03 ± 0.309 µg Hg.g-1 dry wt), or methylmercury (4.11 ± 0.146 µg Hg.g-1 dry wt) diet. After 78 days of feeding, total mercury was highest in the carcass of zebrafish fed methylmercury (12.49 ± 0.369 µg Hg.g-1 dry wt), intermediate in those fed inorganic mercury (1.09 ± 0.117 µg Hg.g-1 dry wt), and lowest in fish fed the control diet (0.48 ± 0.038 µg Hg.g-1 dry wt). Total mercury was ...
Contributing Partner: UNT Libraries
Endocannabinoid System in a Planarian Model

Endocannabinoid System in a Planarian Model

Date: December 2010
Creator: Mustonen, Katie Lynn
Description: In this study, the presence and possible function of endocannabinoid ligands in the planarian is investigated. The endocannabinoids ananadamide (AEA) and 2-arachidonoylglycerol (2-AG) and entourage NAE compounds palmitoylethanolamide (PEA), stearoylethanolamide (SEA) and oleoylethanolamide (OEA) were found in Dugesia dorotocephala. Changes in SEA, PEA, and AEA levels were observed over the initial twelve hours of active regeneration. Exogenously applied AEA, 2-AG and their catabolic inhibition effected biphasic changes in locomotor velocity, analogous to those observed in murines. The genome of a close relative, Schmidtea mediterranea, courtesy of the University of Utah S. med genome database, was explored for cannabinoid receptors, none were found. A putative fatty acid amide hydrolase (FAAH) homolog was found in Schmidtea mediterranea.
Contributing Partner: UNT Libraries
Analysis of Trace Amounts of Adulterants Found in Powders/Supplements Utilizing Direct Inject, Nanomanipulation, and Mass Spectrometry

Analysis of Trace Amounts of Adulterants Found in Powders/Supplements Utilizing Direct Inject, Nanomanipulation, and Mass Spectrometry

Date: August 2016
Creator: Nnaji, Chinyere
Description: The regulations of many food products in the United States have been made and followed very well but unfortunately some products are not put under such rigorous standards as others. This leads to products being sold, that are thought to be healthy, but in reality contain unknown ingredients that may be hazardous to the consumers. With the use of several instrumentations and techniques the detection, characterization and identification of these unknown contaminates can be determined. Both the AZ-100 and the TE2000 inverted microscope were used for visual characterizations, image collection and to help guide the extraction. Direct analyte-probed nanoextraction (DAPNe) technique and nanospray ionization mass spectrometry (NSI-MS) was the technique used for examination and identification of all adulterants. A Raman imaging technique was than introduced and has proven to be a rapid, non-destructive and distinctive way to localize a specific adulterant. By compiling these techniques then applying them to the FDA supplied test samples three major adulterants were detected and identified.
Contributing Partner: UNT Libraries
The Microbial Retting Environment of Hibiscus Cannabinus and Its Implications in Broader Applications

The Microbial Retting Environment of Hibiscus Cannabinus and Its Implications in Broader Applications

Date: May 2015
Creator: Visi, David K.
Description: Fiber-yielding plants is an area of increased interest due to the potential use in a variety of green-based materials. These biocomposites can be incorporated into multiple uses; for example, to replace building materials and interior vehicular paneling. The research here aims to focus in on the crop Hibiscus cannabinus for utilization into these functions. H. cannabinus is economically attractive due to the entire process being able to be accomplished here in the United States. The plant can be grown in a relatively short growth period (120-180 days), and then processed and incorporated in a biocomposite. The plant fiber must first be broken down into a useable medium. This is accomplished by the retting process, which occurs when microbial constituents breakdown the heteropolysaccharides releasing the fiber. The research aims to bridge the gap between the primitive process of retting and current techniques in molecular and microbiology. Utilizing a classical microbiological approach, which entailed enrichment and isolation of pectinase-producing bacteria for downstream use in augmented microbial retting experiments. The tracking of the bacteria was accomplished by using the 16S rRNA which acts as “barcodes” for bacteria. Next-generation sequencing can then provide data from each environment telling the composition and microbial diversity of ...
Contributing Partner: UNT Libraries
A Comparison of Mercury Localization, Speciation, and Histology in Multiple Fish Species From Caddo Lake, a Fresh Water Wetland

A Comparison of Mercury Localization, Speciation, and Histology in Multiple Fish Species From Caddo Lake, a Fresh Water Wetland

Date: May 2012
Creator: Smith, James Durward
Description: This work explores the metabolism of mercury in liver and spleen tissue of fish from a methylmercury contaminated wetland. Wild-caught bass, catfish, bowfin and gar were collected. Macrophage centers, which are both reactive and primary germinal centers in various fish tissues, were hypothesized to be the cause of demethylation of methylmercury in fish tissue. Macrophage centers are differentially expressed in fish tissue based on phylogenetic lineage, and are found primarily in the livers of preteleostean fish and in the spleen of teleostean fish. Histology of liver and spleen was examined in both control and wild-caught fish for pathology, size and number of macrophage centers, and for localization of mercury. Total mercury was estimated in the muscle tissue of all fish by direct mercury analysis. Selenium and mercury concentrations were examined in the livers of wild-caught fish by liquid introduction inductively coupled plasma mass spectrometry (ICP-MS). Total mercury was localized in histologic sections by laser ablation ICP-MS (LA-ICP-MS). Mercury speciation was determined for inorganic and methylmercury in liver and spleen of fish by bas chromatography-cold vapor atomic fluorescence spectroscopy (GC-CVAFS). Macrophage center tissue distribution was found to be consistent with the literature, with a predominance of centers in preteleostean liver and ...
Contributing Partner: UNT Libraries
Effects of Layer Double Hydroxide Nanoclays on the Toxicity of Copper to Daphnia Magna

Effects of Layer Double Hydroxide Nanoclays on the Toxicity of Copper to Daphnia Magna

Date: May 2012
Creator: Blake, Deanne Renee
Description: Nanoparticles may affect secondary pollutants such as copper. Layer Double Hydroxides (LDH) are synthetically produced nanoparticles that adsorb copper via cation exchange. Pretreatment of copper test solutions with LDH nanoparticles followed by filtration removal of LDH nanoparticles demonstrated the smallest LDH aggregates removed the most copper toxicity. This was due to increased surface area for cation exchange relative to larger particle aggregates. Co-exposure tests of copper chloride and clay were run to determine if smaller clay particles increased copper uptake by D. magna. Coexposure treatments had lower LC50 values compared to the filtration tests, likely as a result of additive toxicity. LDH nanoclays do reduce copper toxicity in Daphnia magna and may serve as a remediation tool.
Contributing Partner: UNT Libraries
Investigation of the Pharmacokinetics of Diazepam in Juvenile Channel Catfish (Ictalurus Punctatus)

Investigation of the Pharmacokinetics of Diazepam in Juvenile Channel Catfish (Ictalurus Punctatus)

Date: December 2013
Creator: Overturf, Carmen L.
Description: The presence of pharmaceuticals in the environment is becoming an increasing regulatory and scientific concern. Thus, the metabolic profile and bioconcentration potential of diazepam, a model benzodiazepine, were examined, as well as effects on the endocrine system in channel catfish. Through the use of specific and non-specific cytochrome P450 (CYP450) inhibitors, it was determined that CYP3A-like enzymes may play a role in the biotransformation of diazepam into temazepam; however, the isoform(s) required for the formation of other metabolites is still unknown. Overall, only around 7-8% of diazepam is biotransformed into two known metabolites. Due to the lack of inherent metabolism of diazepam in channel catfish, further analysis was conducted to determine the tissue-specific bioconcentration potential of diazepam in catfish. Various tissues were analyzed for the presence of diazepam as well as metabolites and bioconcentration factors (BCF) were calculated, which were all well below regulatory threshold values (> 2000). Additionally, modulation of the endocrine system by diazepam was examined by measuring steroid hormone concentrations and analyzing mRNA expression of selected steroidogenic enzymes and receptors. Two steroidogenic enzymes were modulated following diazepam exposure, indicating potential endocrine disrupting properties of diazepam. Together, these data suggest that diazepam exhibits low metabolic transformation rates in ...
Contributing Partner: UNT Libraries
Photo-induced Toxicity of Deepwater Horizon Spill Oil to Four Native Gulf of Mexico Species

Photo-induced Toxicity of Deepwater Horizon Spill Oil to Four Native Gulf of Mexico Species

Date: December 2015
Creator: Alloy, Matthew M.
Description: The 2010 Deepwater Horizon oil spill resulted in the accidental release of millions of barrels of crude oil into the Gulf of Mexico (GoM). Photo-induced toxicity following co-exposure to ultraviolet (UV) radiation is one mechanism by which polycyclic aromatic hydrocarbons (PAHs) from oil spills may exert toxicity. Blue crab (Callinectes sapidus) are an important commercial and ecological resource in the Gulf of Mexico and their largely transparent larvae may make them sensitive to PAH photo-induced toxicity. Mahi-mahi (Coryphaena hippurus), an important fishery resource, have positively buoyant, transparent eggs. These characteristics may result in mahi-mahi embryos being at particular risk from photo-induced toxicity. Red drum (Sciaenops ocellatus) and speckled seatrout (Cynoscion nebulosus) are both important fishery resources in the GoM. They spawn near-shore and produce positively buoyant embryos that hatch into larvae in about 24 h. The goal of this body of work was to determine whether exposure to UV as natural sunlight enhances the toxicity of crude oil to early lifestage GoM species. Larval and embryonic organisms were exposed to several dilutions of water accommodated fractions (WAF) from several different oils collected in the field under chain of custody during the 2010 spill and two to three gradations of natural ...
Contributing Partner: UNT Libraries
Dynamics of Stream Fish Metacommunities in Response to Drought and Reconnectivity

Dynamics of Stream Fish Metacommunities in Response to Drought and Reconnectivity

Date: August 2015
Creator: Driver, Lucas J.
Description: This dissertation investigates the spatio-temporal dynamics of intermittent stream fish metacommunities in response drought-induced fragmentation and re-connectivity using both field and experimental approaches. A detailed field study was conducted in two streams and included pre-drought, drought, and post-drought hydrological periods. Fish assemblages and metacommunity structure responded strongly to changes in hydrological conditions with dramatic declines in species richness and abundance during prolonged drought. Return of stream flows resulted in a trend toward recovery but ultimately assemblages failed to fully recover. Differential mortality, dispersal, recruitment among species indicates species specific responses to hydrologic fragmentation, connectivity, and habitat refugia. Two manipulative experiments tested the effects of drought conditions on realistic fish assemblages. Fishes responded strongly to drought conditions in which deeper pools acted as refugia, harboring greater numbers of fish. Variability in assemblage structure and movement patterns among stream pools indicated species specific habitat preferences in response predation, resource competition, and desiccation. Connecting stream flows mediated the impacts of drought conditions and metacommunity dynamics in both experiments. Results from field and experimental studies indicate that stream fish metacommunities are influenced by changes in hydrological conditions and that the timing, duration, and magnitude of drought-induced fragmentation and reconnecting stream flows have important consequences ...
Contributing Partner: UNT Libraries
Ultraviolet Radiation Tolerance in High Elevation Copepods from the Rocky Mountains of Colorado, USA

Ultraviolet Radiation Tolerance in High Elevation Copepods from the Rocky Mountains of Colorado, USA

Date: December 2011
Creator: Hudelson, Karista
Description: Copepods in high elevation lakes and ponds in Colorado are exposed to significant levels of ultraviolet radiation (UV), necessitating development of UV avoidance behavior and photoprotective physiological adaptations. The copepods are brightly pigmented due to accumulation of astaxanthin, a carotenoid which has photoprotective and antioxidant properties. Astaxanthin interacts with a crustacyanin-like protein, shifting its absorbance from 473 nm (hydrophobic free form, appears red) to 632 nm (protein-bound complex, appears blue). In six sites in Colorado, habitat-specific coloration patterns related to carotenoprotein complex have been observed. The objective of this study was to determine whether pigment accumulation or carotenoprotein expression has a greater effect on resistance to UV exposure. For each site, copepod tolerance to UV was assessed by survivorship during UV exposure trials. Average UV exposure was determined for each habitat. Astaxanthin profiles were generated for copepods in each site. Ability to withstand UV exposure during exposure trials was significantly different between color morphs (p < 0.0001). Red copepods were found to tolerate 2-fold greater levels of UVB than blue or mixed copepods. Additionally, red copepods have much higher levels of total astaxanthin than blue or mixed copepods (p < 0.0001) and receive a higher daily UV dose (p < ...
Contributing Partner: UNT Libraries
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 ...
Contributing Partner: UNT Libraries
Effects of Suspended Multi-Walled Carbon Nanotubes on Daphnid Growth and Reproduction

Effects of Suspended Multi-Walled Carbon Nanotubes on Daphnid Growth and Reproduction

Date: May 2010
Creator: Alloy, Matthew Michael
Description: Multi-walled carbon nanotube aggregates can be suspended in the aqueous phase by natural organic matter. These aggregates are ingested by filter feeding zooplankton. Ingested aggregates result in decreased growth and decreased reproduction. These effects may be caused by reduction in energy input from normal feeding behavior. pH alters natural organic matter structure through changes in electrostatic repulsion. Altered natural organic matter structure changes multi-walled carbon nanotube aggregate size. This size variation with variation in pH is significant, but not large enough a change in size to alter toxicity, as the aggregate size range remains well within the particle size selection of the organisms.
Contributing Partner: UNT Libraries
Glucose Induces Sensitivity to Oxygen Deprivation and Alters Gene Expression in Caenorhabditis Elegans

Glucose Induces Sensitivity to Oxygen Deprivation and Alters Gene Expression in Caenorhabditis Elegans

Date: August 2015
Creator: Garcia, Anastacia M.
Description: An organisms’ diet represents an exogenous influence that often yields colossal effects on long-term health and disease risk. The overconsumption of dietary sugars for example, has contributed to significant increases in obesity and type-2 diabetes; health issues that are costly both economically and in terms of human life. Individuals who are obese or are type-2 diabetic often have compromised oxygen delivery and an increased vulnerability to oxygen-deprivation related complications, such as ischemic strokes, peripheral arterial disease and myocardial infarction. Thus, it is of interest to identify the molecular changes glucose supplementation or hyperglycemia can induce, which ultimately compromise oxygen deprivation responses. By utilizing the Caenorhabditis elegans genetic model system, which is anoxia tolerant, I determined that a glucose-supplemented diet negatively impacts responses to anoxia and that the insulin-like signaling pathway, through fatty acid and ceramide biosynthesis and antioxidant activity, modulates anoxia survival. Additionally, a glucose-supplemented diet induces lipid accumulation. Use of RNA-sequencing analysis to compare gene expression responses in animals fed either a standard or glucose-supplemented diet revealed that glucose impacts the expression of genes involved with multiple cellular processes including lipid and carbohydrate metabolism, stress responses, cell division, and extracellular functions. Several of the genes we identified are homologous ...
Contributing Partner: UNT Libraries
Stress Response by Alternative Σ-factor, Rpoh, and Analysis of Posttranslational Modification of the Heat Shock Protein, Dnak, in Escherichia Coli

Stress Response by Alternative Σ-factor, Rpoh, and Analysis of Posttranslational Modification of the Heat Shock Protein, Dnak, in Escherichia Coli

Date: May 2015
Creator: Martinez, Sarah N
Description: Bacteria have developed specialized responses that involve the expression of particular genes present in a given regulon. Sigma factors provide regulatory mechanisms to respond to stress by acting as transcriptional initiation factors. This work focuses on σ32 during oxidative stress in Escherichia coli. The differential response of key heat shock (HS) genes was investigated during HS and oxidative stress using qPCR techniques. While groEL and dnaJ experienced increases in transcriptional response to H2O2 (10 mM), HS (42°C), and paraquat (50 mM) exposure, the abundance of dnaK over the co-chaperones was apparent. It was hypothesized that DnaK undergoes oxidative modification by reactive carbonyls at its Lys-rich C-terminus, accounting for the differential response during oxidative stress. A σ32-mediated β-galactosidase reporter was devised to detect the activity of wild-type DnaK and DnaKV634X modified to lack the Lys-rich C-terminus. Under unstressed conditions and HS, σ32 was bound at the same rate in both strains. When subjected to H2O2, the WT DnaK strain produced significantly higher β-galactosidase than DnaKV634X (one-tailed Student’s t test p=0.000002, α=0.05) and approached the same level of output as the lacZ positive control. The β-galactosidase assay indicates that DnaK undergoes Lys modification in the WT strain, preventing the protein from binding ...
Contributing Partner: UNT Libraries
Determination of Bioconcentration Potential of Selected Pharmaceuticals in Fathead Minnow, Pimephales promelas, and Channel Catfish, Ictalurus punctatus

Determination of Bioconcentration Potential of Selected Pharmaceuticals in Fathead Minnow, Pimephales promelas, and Channel Catfish, Ictalurus punctatus

Date: December 2010
Creator: Nallani, Gopinath C.
Description: The primary objective of this work was to determine the tissue-specific bioconcentration factors (BCFs) of the selected pharmaceuticals: norethindrone (NET), ibuprofen (IBU), verapamil (VER), clozapine (CLZ) and fenofibrate (FFB) in two freshwater fishes: fathead minnow and channel catfish. BCF tests on fathead followed the standard OECD 42-day test while a 14-day abridged test design was used in catfish exposures. Additional objectives included a) comparing the measured BCFs to the US EPA's BCFWIN model predicted values, b) comparing the BCF results from the standard and reduced tests, and c) prediction of chronic risk of the pharmaceuticals in fish using the human therapeutic plasma concentrations. Each test included uptake and depuration phases to measure tissue-specific kinetic BCFs. The results indicated that all the pharmaceuticals, except IBU, have the potential for accumulation in fish. Estimated BCFs for NET, VER and FFB may not be significant in view of the current regulatory trigger level (BCF &#8805; 2000); however, CLZ's BCF in the liver had approached the criterion level. Significant differences were noticed in the tissue-specific uptake levels of the pharmaceuticals with the following general trend: (liver/kidney) > (gill/brain) > (heart/muscle) > plasma. IBU uptake was highest in the plasma. When compared to the measured ...
Contributing Partner: UNT Libraries