UNT Theses and Dissertations - 5 Matching Results

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Carbachol- and ACPD-Induced Phosphoinositide Responses in the Developing Rat Neocortex

Description: Signal transduction via the phosphoinositide (PI) second messenger system has key roles in the development and plasticity of the neocortex. The present study localized PI responses to individual cortical layers in slices of developing rat somatosensory cortex. The acetylcholine agonist carbachol and the glutamate agonist trans-1-amino-1,3-cyclopentanedicarboxylic acid (ACPD) were used to stimulate PI turnover. The PI responses were compared to the distribution of the corresponding PI-linked receptors in order to investigate the regional ontogeny of PI coupling to receptors in relation to neural development. The method for assessing PI turnover was modified from Hwang et al. (1990). This method images the PI response autoradiographically through the localizaton of [3H]cytidine that has been incorporated into the membrane-bound intermediate, cytidine diphosphate diacylglycerol. In each age group (postnatal days 4-30), carbachol resulted in more overall labeling than ACPD. For both agonists, the response peaked on postnatal day 10 (P10) and was lowest in the oldest age group. The laminar distribution of the carbachol PI response from P4-P16 corresponded fairly well with the laminar distribution of [3H]quinuclidinyl benzilate binding (Fuchs, 1995). However, in the subplate layer the carbachol response was strong while receptor binding was minimal. The carbachol response decreased after postnatal day 10, while the overall levels of receptor binding continued to increase. From P5 - P14, PI-linked metabotropic glutamate receptors are most concentrated in layer IV (Blue et al., 1997), whereas only on P6 was there a correspondingly high ACPD-initiated PI response in this layer. Unlike receptors, the PI response was strong in upper V (P4 - P12) and within layers II/III (P8 - P16). From P4 - P21, the subplate showed relatively high PI labeling compared to receptor binding. The several differences between the distribution of PI response and receptors suggest spatiotemporal heterogeneity of receptor coupling to second messenger systems.
Date: August 2000
Creator: Hartgraves, Morri D.
Partner: UNT Libraries

BioInformatics, Phylogenetics, and Aspartate Transcarbamoylase

Description: In this research, the necessity of understanding and using bioinformatics is demonstrated using the enzyme aspartate transcarbamoylase (ATCase) as the model enzyme. The first portion of this research focuses on the use of bioinformatics. A partial sequence of the pyrB gene found in Enterococcus faecalis was submitted to GenBank and was analyzed against the contiguous sequence from its own genome project. A BLAST (Basic Local Alignment Search Tool; Atschul, et al., 1990) was performed in order to hypothesize the remaining portion of the gene from the contiguous sequence. This allowed a global comparison to other known aspartate transcarbamoylases (ATCases) and once deduced, a translation of the sequence gave the stop codon and thus the complete sequence of the open reading frame. When this was complete, upstream and downstream primers were designed in order to amplify the gene from genomic DNA. The amplified product was then sequenced and used later in phylogenetic analyses concerning the evolution of ATCase. The second portion of this research involves taking multiple ATCase nucleotide sequences and performing phenetic and phylogenetic analyses of the archaea and eubacter families. From these analyses, ancestral relationships which dictate both structure and function were extrapolated from the data and discussed.
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Date: August 2000
Creator: Cooke, Patrick Alan
Partner: UNT Libraries

Comparative Biochemistry and Evolution of Aspartate Transcarbamoylase from Diverse Bacteria

Description: Aspartate transcarbamoylase (ATCase) catalyzes the first committed step in pyrimidine biosynthesis. Bacterial ATCases are divided into three classes, A, B and C. Class A ATCases are largest at 450-500, are. dodecamers and represented by Pseudomonas ATCase. The overlapping pyrBC' genes encode the Pseudomonases ATCase, which is active only as a 480 kDa dodecamer and requires an inactive pyrC'-encoded DHOase for ATCase activity. ATCase has been studied in two non-pathogenic members of Mycobacterium, M. smegmatis and M. phlei. Their ATCases are dodecamers of molecular weight 480 kDa, composed of six PyrB and six PyrC polypeptides. Unlike the Pseudomonas ATCase, the PyrC polypeptide in these mycobacteria encodes an active DHOase. Moreover, the ATCase: DHOase complex in M. smegmatis is active both as the native 480 kDa and as a 390 kDa complex. The latter lacks two PyrC polypeptides yet retains ATCase activity. The ATCase from M. phlei is similar, except that it is active as the native 480 kDa form but also as 450,410 and 380 kDa forms. These complexes lack one, two, and three PyrC polypeptides, respectively. By contrast,.ATCases from pathogenic mycobacteria are active only at 480 kDa. Mycobacterial ATCases contain active DHOases and accordingly. are placed in class A1 . The class A1 ATCases contain active DHOases while class A2 ATCases contain inactive DHOases. ATCase has also been purified from Burkholderia cepacia and from an E. coli strain in which the cloned pyrB of B. cepacia was expressed. The B. cepacia ATCase has a molecular mass of 550 kDa, with two different polypeptides, PyrB (52 kDa) and PyrC of (39 kDa). The enzyme is active both as the native enzyme at 550 kDa and as smaller molecular forms including 240 kDa and 165 kDa. The ATCase synthesized by the cloned pyrB gene has a molecular weight of 165 kDa composed ...
Date: May 1999
Creator: Hooshdaran, Massoumeh Ziba
Partner: UNT Libraries

L-asparaginase II Production by Escherichia coli

Description: Growth of Escherichia coli A-l under aerobic conditions in an enriched medium with a total amount of 0.2 per cent glucose was biphasic and asparaginase II activity was detected after depletion of ammonia from the growth medium in the second phase of growth. Glucose was exhausted two hours before ammonia and three hours before asparaginase II activity was detected. The concentration of 3',5'-cyclic adenosine monophosphate was found to fluctuate when the dissolved oxygen in the medium reached a low level, when glucose and ammonia were exhausted, and when the cells entered the second stationary phase of growth. Culture tube studies of the growth of E_j_ coli A-l in three per cent nutrient broth with varied concentrations of ammonium chloride and potassium nitrate gave lower specific activity of asparaginase II when this was compared to that seen in three per cent nutrient broth alone. The addition of glucose to the same medium before asparaginase II activity was detected resulted in the production of acid by E. coli A-l with cessation of growth; however, addition after L-asparaginase synthesis had started did not affect the specific activity of the enzyme. The addition of ammonium chloride suppressed L-asparaginase synthesis, but addition after enzyme synthesis started had no affect. These findings suggest that asparaginase II is produced by E. coli A-l in response to low concentrations of ammonia and that exogenously supplied nitrogen compounds may play a major role in the regulation of this enzyme. It is suggested that E. coli A-l produced L-asparaginase in order to obtain ammonia for the synthesis of glutamine from glutamate. The synthesis of glutamine from glutamate is the first step of a highly branched pathway which ultimately leads to the synthesis of many of the important macromolecules of the cell.
Date: May 1985
Creator: Johnson, Terrance L. (Terrance Lewyne), 1950-
Partner: UNT Libraries

Degradation of Humic Substances by Aquatic Bacteria

Description: A variety of aquatic bacteria were isolated and tested for their ability to degrade humic substances and their aromatic residues/monomers which serve as precursors of the trihalomethanes (THMs) found in chlorinated drinking waters. The majority of them were Gram-negative, oxidative types dominated by pseudomonads. Most of the 146 isolates were found to utilize as their sole source of carbon several or more of ten aromatic compounds known to be products of degradation of humus and also to be precursors of THMs. The aromatics tested, with percent of the isolates utilizing the compound in parentheses, were: p-hydroxybenzoate (49), vanillic acid (48), 3,5-dihydroxybenzoic acid (16), syringic acid (19), vanillin (30), benzoic acid (27), ferulic acid (34), resorcinol (9), catechol (8) and protocatechuic acid (27).
Date: August 1985
Creator: Baiu, Saleh Hamed Salem
Partner: UNT Libraries