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 Department: Department of Biological Sciences
 Collection: UNT Theses and Dissertations
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 ...
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Origin and Role of Factor Viia

Origin and Role of Factor Viia

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Date: December 2013
Creator: Khandekar, Gauri
Description: Factor VII, the initiator of the extrinsic coagulation cascade, circulates in human plasma mainly in its zymogen form, Factor VII and in small amounts in its activated form, Factor VIIa. However, the mechanism of initial generation of Factor VIIa is not known despite intensive research using currently available model systems. Earlier findings suggested serine proteases Factor VII activating protease, and hepsin play a role in activating Factor VII, however, it has remained controversial. In this work I estimated the levels of Factor VIIa and Factor VII for the first time in adult zebrafish plasma and also reevaluated the role of the above two serine proteases in activating Factor VII in vivo using zebrafish as a model system. Knockdown of factor VII activating protease did not reduce Factor VIIa levels while hepsin knockdown reduced Factor VIIa levels. After identifying role of hepsin in Factor VII activation in zebrafish, I wanted to identify novel serine proteases playing a role in Factor VII activation. However, a large scale knockdown of all serine proteases in zebrafish genome using available knockdown techniques is prohibitively expensive. Hence, I developed an inexpensive gene knockdown method which was validated with IIb gene knockdown, and knockdown all serine proteases ...
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Designing Tools to Probe the Calcium-dependent Function of Arabidopsis Tonneau2

Designing Tools to Probe the Calcium-dependent Function of Arabidopsis Tonneau2

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Date: December 2013
Creator: Oremade, Oladapo O.
Description: Plants possess unique features in many aspects of development. One of these features is seen in cell wall placement during cytokinesis, which is determined by the position of the preprophase band (PPB) and the subsequent expansion of the phragmoplast that deposits the new cell wall. During phragmoplast expansion, the phragmoplast tracks to the cortical division site, which was delineated by the PPB. Thus the position of the PPB determines the orientation of the division plane. In Arabidopsis thaliana, TONNEAU2 (TON2) is required for PPB formation and has been shown to interact with a type A subunit of the PP2A phosphatase in the yeast two-hybrid system. In Arabidopsis tonneau2 (ton2) mutants, abnormalities of the cortical microtubule cytoskeleton, such as disorganization of the interphase microtubule array and lack of PPB formation before mitosis markedly affects cell shape and arrangement as well as overall plant morphology. Loss of dcd1/add1, the maize ton2 homologues gives rise to a similar phenotype in Zea mays. The TON2 protein has two EF hand domains which are calcium-binding sites. Since calcium has been known to play key roles in several areas of plant functioning, the following question was raised: “Does calcium binding contribute to the localization and function ...
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Field and Laboratory Fish Tissue Accumulation of Carbamazepine and Amiodarone

Field and Laboratory Fish Tissue Accumulation of Carbamazepine and Amiodarone

Date: December 2013
Creator: García Martínez, Santos Noé
Description: The goals of this dissertation work were to assess the bioaccumulation potential of carbamazepine and amiodarone, two widely used ionizable pharmaceutical compounds that possess mid-range and high LogD values, respectively, and to evaluate alternative methods to assess chemical accumulation in bluntnose minnows, catfish, and tilapia. Results indicated that carbamazepine does not appreciably bioaccumulate in fish tissue with BCFk and BAF carbamazepine values < 10. Amiodarone, however, with a log D of 5.87 at pH 7.4, accumulated in fish tissues with kinetic BCF values <2,400. Collectively, the data suggest that full and abbreviated laboratory-derived BCFs, BCFMs derived from S9 loss-of-parent assays, as well as field BAF values are similar for each of the two drugs. In summary, the results from this dissertation indicated: 1) The reduced design BCF test is a good estimate for the traditional OECD 305 test. 2) In vitro S9 metabolism assays provide comparable BCF estimates to the OECD 305 test. 3) Metabolism may play a large role in the accumulation of drugs in fish. 4) Reduced BCF tests and in vitro assays are cost effective and can reduce vertebrate testing.
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Functional Characterization of Mtnip/latd’s Biochemical and Biological Function

Functional Characterization of Mtnip/latd’s Biochemical and Biological Function

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Date: December 2013
Creator: Bagchi, Rammyani
Description: Symbiotic nitrogen fixation occurs in plants harboring nitrogen-fixing bacteria within the plant tissue. The most widely studied association is between the legumes and rhizobia. In this relationship the plant (legumes) provides the bacteria (rhizobia) with reduced carbon derived from photosynthesis in exchange for reduced atmospheric nitrogen. This allows the plant to survive in soil, which is low in available of nitrogen. Rhizobia infect and enter plant root and reside in organs known as nodules. In the nodules the bacteria fix atmospheric nitrogen. The association between the legume, Medicago truncatula and the bacteria Sinorhizobium meliloti, has been studied in detail. Medicago mutants that have defects in nodulation help us understand the process of nitrogen fixation better. One such mutant is the Mtnip-1. Mtnip-1 plants respond to S. meliloti by producing abnormal nodules in which numerous aberrant infection threads are produced, with very rare rhizobial release into host plant cells. The mutant plant Mtnip-1 has an abnormal defense-like response in root nodules as well as defects in lateral root development. Three alleles of the Mtnip/latd mutants, Mtnip-1, Mtlatd and Mtnip-3 show different degrees of severity in their phenotype. Phylogenetic analysis showed that MtNIP/LATD encodes a protein belonging to the NRT1(PTR) family of ...
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The Stoneflies (Plecoptera) of the Ozark and Ouachita Mountains

The Stoneflies (Plecoptera) of the Ozark and Ouachita Mountains

Date: December 1989
Creator: Poulton, Barry C. (Barry Charles)
Description: Collections of stoneflies (Plecoptera) were made at 603 stream sites from Nov. 1983 - May 1988 in the Ozark-Ouachita Mountain region, in relation to physiographic and vegetational characteristics. Examination of approximately 9000 vials from these collections, supplemented with material from major museums and other collectors, revealed 88 stonefly species in 8 families and 24 genera. Pearson's measure of association (R) showed there was a significant association between species present and each of the tested variables.
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Pyrimidine Metabolism in Rhizobium: Physiological Aspects of Pyrimidine Salvage

Pyrimidine Metabolism in Rhizobium: Physiological Aspects of Pyrimidine Salvage

Date: December 1989
Creator: Ibrahim, Mohamed M.
Description: The objective of this research was to study the pyrimidine salvage pathways of Rhizobium. Three approaches were used to define the pyrimidine salvage pathways operative in two species of Rhizobium, R. meliloti and R. leguminosarum . The first approach was to ascertain the pyrimidine bases and nucleosides that could satisfy the pyrimidine requirement of pyrimidine auxotrophs. Uracil, cytosine, uridine or cytidine all satisfied the absolute pyrimidine requirement. The second approach was to select for mutants resistant to 5-fluoropyrimidine analogues which block known steps in the interconversion of the pyrimidine bases and nucleosides. Mutants resistant to 5-fluorouracil lacked the enzyme uracil phosphoribosyltransferase (upp ) and could no longer use uracil to satisfy their pyrimidine requirement. Mutants resistant to 5-fluorocytosine, while remaining sensitive to 5- fluorouracil, lacked cytosine deaminase (cod) and thus could no longer use cytosine to satisfy their pyrimidine auxotrophy. The third approach used a reversed phase HPLC column to identify the products that accumulated when cytidine, uridine or cytosine was incubated with cell extracts of wild type and analogue resistant mutants of Rhizobium. When cytidine was incubated with cell extracts of Rhizobium wild type, uridine, uracil and cytosine were produced. This Indicated that Rhizobium had an active cytidine deaminase ...
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Measurement of Feedback Inhibition In Vivo and Selection of ATCase Feedback Altered Mutants in Salmonella typhimurium

Measurement of Feedback Inhibition In Vivo and Selection of ATCase Feedback Altered Mutants in Salmonella typhimurium

Date: August 1989
Creator: Bailey, Andrea J., 1952-
Description: Aspartate transcarbamoylase (ATCase; encoded by pyrBI genes) is one of the most studied regulatory enzymes in bacteria. It is feedback inhibited by cytidine triphosphate (CTP) and activated by adenosine triphosphate (ATP). Much is known about the catalytic site of the enzyme, not nearly as much about the regulatory site, to which CTP binds. Until now a positive selection for feedback-modified mutants was not available. The selection we have developed involves the use of a pyrA deletion in S. typhimurium. This strain lacks carbamoylphosphate and requires both a pyrimidine and arginine for growth. In this strain citrulline is used to satisfy the pyrimidine and arginine requirements. The minimal flow through the pyrimidine pathway from the citrulline-produced carbamoylphosphate is exquisitely sensitive to feedback control of ATCase by CTP. By elevating the CTP pool, via exogenous cytidine, in a strain that also contains a cytidine deaminase mutant (cdd) growth can be stopped completely, indicating 100% inhibition. It was therefore possible to measure in vivo feedback inhibition of ATCase among the citrulline users and to isolate a family of ATCase regulatory mutants with either modified or no response to effectors.
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Validation of a Coupled Herbicide Fate and Target Plant Species Effects Model

Validation of a Coupled Herbicide Fate and Target Plant Species Effects Model

Date: December 1989
Creator: Clifford, Philip A. (Philip Alan)
Description: A series of experiments provided data to parameterize and validate a coupled herbicide fate and target plant species effects model. This simulation model is currently designed to predict responses of water hyacinth populations to treatments of the dimethylamine formulation of 2,4- dichloro-phenoxy acetic acid (2,4-D -DMA). Experiments investigated 1) the response of water hyacinth to varying exposures of 2,4-D (DMA); 2) the role of water hyacinth density and herbicide interception in treatment effectiveness using 2,4-D (DMA); and 3) the importance of root exposure to obtain control of water hyacinth using 2,4- D (DMA). Results demonstrated the importance of leaf or canopy interception of 2,4-D (DMA) sprays in obtaining control of water hyacinth populations. The critical threshold plant tissue concentration of 2,4-D (DMA) required to elicit maximum mortality (98%) was estimated to be approximately 12 mg 2,4-D per kg water hyacinth tissue (wet weight). Root uptake apparently plays little or no role in the effectiveness of this herbicide for controlling water hyacinth growth. Validation trials illustrated the efficacy of the current model. The model was validated with data from a field operation. This research has provided considerable insight into optimal use of this auxin-type herbicide for control of water hyacinth, a ...
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Toxicological Characterization of Trinity River Sediments

Toxicological Characterization of Trinity River Sediments

Date: December 1989
Creator: Hall, Jerry F. (Jerry Fowler)
Description: Sediments in the Trinity River were chemically, physically and biologically characterized and assessed for toxicity. Laboratory bioassays were conducted to identify sediments which induced toxic responses in test organisms and to document these responses through time. Metal and organic contaminant concentrations in bottom sediments were measured. Relationships between these concentrations and biological responses observed in laboratory bioassays were determined. Toxicity identification / reduction methods were used to characterize sediment toxicants. Sediment oxygen demand was also measured in resuspended and undisturbed bottom sediments through time. The Background Sediment Chemistry Approach and the Sediment Bioassay Approach were used to assess sediment quality. Sediment toxicity was observed in whole sediment bioassays using Chironomus tentans as the test species. A relationship between sediment contaminant concentration and toxicity was observed in approximately sixty percent of the sediments. Oxygen demand of resuspended sediments was elevated in sediments at two locations on the river. Oxygen demand of undisturbed sediments was elevated at one location on the river. Characterization of sediment toxicants was conducted using EDTA, pH, and carbon treatments and manipulations of the sediments. Aeration tests were also used to evaluate the contribution of volatile organic contaminants to observed toxicity.
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