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Opioid/Adrenergic Interaction in Regulating Canine Cardiac Function

Description: Opioid/adrenergic interactions were studied to evaluate two hypotheses: (1) naloxone potentiates the effect of epinephrine on cardiac contractility by increasing circulating epinephrine concentrations; and (2) endogenous and exogenous opioids alter left cardiac nerve stimulationinduced norepinephrine release and cardiac function. A canine isolated heart-lung preparation was used for the first study. Plasma epinephrine was determined and myocardial epinephrine uptake was calculated during intravenous epinephrine infusion. Naloxone (4 mg) was given and the epinephrine infusion was repeated. Naloxone increased cardiac contractility, coronary blood flow, and the coronary sinus epinephrine concentration. When coronary blood flow was subsequently held constant (100% above resting), naloxone increased only contractility. This result indicated that the previously observed increase in coronary sinus epinephrine was flow dependent. Corticosterone (an uptake II blocker) was employed as a positive control. Corticosterone increased the contractile response to epinephrine, but unlike naloxone, corticosterone was accompanied by a clear decrease in myocardial epinephrine uptake. The stereospecificity of the response to naloxone was investigated and (+) naloxone equaled or exceeded (-) naloxone in potentiating the inotropic effect of epinephrine. In the second study, the left cardiac nerve was isolated and electrically stimulated in intact dogs. Norepinephrine overflow gradually declined during successive control stimulations. Pretreatment with naloxone (100 Mg/kg) prevented or delayed the decline. An intracoronary dynorphin 1-9 infusion (2 nmol/min/kg for 20 minutes) reduced both norepinephrine overflow and cardiac performance, and both effects were prevented by pretreatment with naloxone (100 /xg/kg) . To summarize, naloxone potentiated the inotropic effect of infused epinephrine without altering circulating epinephrine concentrations or myocardial epinephrine uptake. This effect of naloxone was not stereospecific and probably not mediated through a traditional opiate receptor. Endogenous and exogenous opioids inhibited the left cardiac nerve stimulation-induced norepinephrine overflow, suggesting that opiate receptors may regulate cardiac excitability by modulating norepinephrine release.
Date: May 1990
Creator: Gu, Hong

Regulation of Colony-Stimulating Factor-1 Biosynthesis

Description: Recent studies suggest that synthesis of the Colony-stimulating factor (CSF) is a well regulated process. However, the molecular mechanisms of the signal transduction of the various inducers of CSF such as monokines and lymphokines are not well understood. Using Interleukin 1 (IL-1) stimulation of CSF-1 in the MIA PaCa-2 cell line as a model system, the involvement of G-protein has been studied. The IL-1 induction of CSF-1 synthesis can be inhibited by both Pertussis toxin and Cholera toxin, which are known to modify the Gᵢ and Gₛ proteins respectively, thus activating adenylate cyclase to release more cAMP. The toxin inactivation can be prevented by inhibitors of the ADP-ribosylation such as, benzamide and MBAMG. Addition of dibutyryl-cAMP inhibits the IL-1 induced CSF production. Both Theophylline and Forskolin which increase cAMP by inhibiting phosphodiesterase and stimulating adenylate cyclase respectively, also inhibit CSF-1 production. Results from these studies have shown that cAMP level inversely regulates the biosynthesis of CSF-1. Preincubation of MIA PaCa-2 cells with IL-1 and 5'- guanylylimidodiphosphate (GppNHp) prevents the inhibitory effect of pertussis toxin on CSF-1 production. These data are consistent with the hypothesis that IL-1 binds to its receptor and couples to Gᵢ∝ resulting in the inhibition of adenylate cyclase and reducing cAMP level. Lowering of the' cAMP level leads to the activation of CSF-1 gene expression. The activity of another inducer of CSF-1 production in this system, 12-0-tetradecanoylphorbol-13-acetate (TPA), can be abolished by 1- (5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H-7), which is a specific inhibitor of protein kinase C. However, H-7 failed to inhibit IL-1 stimulated CSF-1 production. Other known activators of protein kinase C namely, Ca²⁺ and L-α-l-oleoyl-2-acetoyl-sn- 3-glycerol (OAG), also increase CSF production. On the other hand, Indomethacin which is known to inhibit prostaglandin E (PGE), stimulates CSF-1 production in MIA PaCa-2 cells. These data suggest that different mechanisms ...
Date: May 1990
Creator: Ku, Chun-Ying

Toxicity of Chromium and Fluoranthene From Aqueous and Sediment Sources to Selected Freshwater Fish

Description: Research efforts in aquatic toxicology have historically centered on the chemical analyses and toxic effects of waters to aquatic organisms. More recently, sediment-source toxicity has been explored, with efforts concentrated on establishing sensitive and accurate methodologies. This study focused on the toxicity of trivalent chromium, hexavalent chromium, and fluoranthene to Pimephales promelas, Ictalurus punctatus, and Lepomis macrochirus. Test fish were exposed to both water-borne and sediment-source toxicants for 96 hours (h) and 30 days (d). A 96-h and 30-d LC50 (mg/L Cr, ug/L Fluoranthene) was determined for each fish species exposed to aqueous toxicants. In addition, 96-h and 30-d LC50s were determined for each fish species from sediment chromium concentrations (mg/kg) and sediment fluoranthene concentrations (ug/kg). Although lethality endpoints were used throughout this research, acute effects other than mortality were determined for Lepomis macrochirus exposed to hexavalent chromium. Lethal toxicity values (96-h and 30-d LC50 and their 95% confidence limits) for trivalent chromium could not be determined since trivalent chromium concentations above 6.0 mg/L could not be obtained at water pHs compatible with these fish species. Trivalent chromium addition to test waters at pHs compatible with fish survival resulted in a chromium precipitate that was not lethal to test fish. In contrast, fathead minnows, channel catfish, and bluegill sunfish exposed to hexavalent chromium in water and sediments experienced mortality. Fathead minnows exposed to fluoranthene in water for 96h demonstrated a maximum mortality of 69%, while 100% mortality was achieved with channel catfish in similar tests. Sediment tests with fluoranthene resulted in 100% mortality with both fathead minnows and channel catfish.
Date: May 1990
Creator: Gendusa, Tony C.

Isolation, Characterization and Physiological Studies of Cyanide-Utilizing Bacteria

Description: Ten bacteria capable of growth on the metal-cyano complex, tetracyanonickelate (II) {K2 [Ni(CN)J } (TCN), supplied as the sole nitrogen source, were isolated. Seven isolates were identified as pseudomonads while the remaining three were classified as Klebsiella species. In addition to TCN, all isolates were able to utilize KCN although it was significantly more toxic. The degradation of TCN was most complete when supplied at growth-limiting concentrations, did not occur when ammonia was present, and resulted in the formation of nickel cyanide [Ni(CN)2] as a degradation product.
Date: December 1991
Creator: Silva Avalos, Juan G. (Juan Guillermo)

Physiological Ecology, Population Genetic Responses and Assemblage Stability of Fishes in Two Southwestern Intermittent Stream Systems

Description: Six sites within the Denton and Hickory Creek watersheds were sampled over three years to assess the impact of seasonal intermittent stream conditions on the ichthyofauna. An integrated approach using field and laboratory techniques was employed to evaluate the responses of the fishes.
Date: December 1991
Creator: Rutledge, Charles Jerry, 1941-

Physiological Effects of Ascaris Suum Intestinal Microflora on 5-Hydroxytryptamine Level and Binding Sites in the Intestinal Epithelial Cells

Description: Serotonin (5-hydroxytryptamine, 5-HT) has been shown to activate carbohydrate metabolism in adult female Ascaris suum. Serotonin may be either absorbed directly from the environment or synthesized de novo from the absorbed L-tryptophan in adult female A. suum. The enzymes necessary for the synthesis of 5-HT have been identified in both intestine and muscle tissues. The serotonin absorbed from the environment is obtained either from the host's gastrointestinal contents or from the 5-HT producing bacteria in the intestine of A. suum. Numerous 5-HT producing bacteria were identified in the intestinal microflora. The physiological contributions of 5-HT producing bacteria to the 5-HT level, turnover and binding sites in the intestinal tissue of A. suum were investigated.
Date: December 1991
Creator: Shahkolahi, Akbar Mohammadpour

Blood Pressure Regulation During Simulated Orthostatism Prior to and Following Endurance Exercise Training

Description: Cardiovascular responses and tolerance to an orthostatic stress were examined in eight men before and after eight months of endurance exercise training. Following training, maximal oxygen consumption and blood volume were increased, and resting heart rate reduced. Orthostatic tolerance was reduced following training in all eight subjects. It was concluded that prolonged endurance training decreased orthostatic tolerance and this decrease in tolerance appeared associated with attenuated baroreflex sensitivity and alterations in autonomic balance secondary to an increased parasympathetic tone noted with training.
Date: May 1992
Creator: Stevens, Glen Harold John

Construction of a Cloning Vector Based upon a Rhizobium Plasmid Origin of Replication and its Application to Genetic Engineering of Rhizobium Strains

Description: Rhizobia are Gram-negative, rod-shaped, soil bacteria with the ability to fix atmospheric nitrogen into ammonia as symbiont bacteroids within nodules of leguminous plant roots. Here, resident Rhizobium plasmids were studied as possible sources of components for the construction of a cloning vector for Rhizobium species.
Date: May 1992
Creator: Jeong, Pyengsoo

Enzyme Assays Using Earthworms for Assessing Innate and Nonspecific Immunotoxicity of Xenobiotics

Description: Principal objectives of my research were to: (1) report for the first time that coelomocytes are able to reduce NBT dye and confirm the presence of lysozyme-like activity in earthworm; (2) develop a standard methodology for determination of NBT reduction and lysozyme-like activity in earthworms; (3) compare NBT reduction and lysozyme-like activity in earthworms with those of murine and human cells and fluids; and (4) demonstrate the sensitivity of earthworm NBT reduction and lysozyme-like activity as the assays using matrics in refuse-derived fuel fly ash (RDFF) and CuSO4.
Date: May 1992
Creator: Chen, Shing-Chong

Molecular and Kinetic Characterization of the Aspartate Transcarbamoylase Dihydroorotase Complex in Pseudomonas putida

Description: Aerobic Gram negative bacteria such as Pseudomonas putida were reported to possess class A ATCases and to have a M.W. of 360 kD. The nucleotide sequence of the P. putida pyrBC was determined to answer this question once and for all. The expected regulatory gene was not found. It is shown that the P. putida pyrB gene is overlapped by pyrC by 4 bp. The P.putida pyrB is 1005 bp (335 aa) in length and the pyrC is 1275 bp (425 aa) long. Both of these genes complement E. coli mutants with their respective genotypes. Another finding borne out from the sequence is an effector binding site at the N-terminus of pyrB of P. putIda. The binding site shows that effectors compete with carbamoylphosphate for the active site. In this dissertation, it is shown that the ATCase of P.putida is a trimer of M.W. of 109 kD (3 x 36.4 kD) and that the gene encoding pyrB is overlapped by the pyrC gene which encodes DHOase. It is also shown that the pyrBC encoded enzymes copurify as a dodecameric complex with a M.W. of 484 kD.
Date: May 1992
Creator: Schurr, Michael J. (Michael John)

Stock and Species Identification of Selected Marine Fishes and Shellfishes Using Allozyme Analysis and Isoelectric Focusing: Implications for Texas Fisheries Management

Description: Allozyme frequencies and general protein patterns were surveyed among selected Texas marine fishes and shellfishes to illustrate the application of biochemical genetic techniques to stock and species identification in fisheries management.
Date: May 1992
Creator: King, Timothy L. (Timothy Lee)

Subcloning and Nucleotide Sequence of Two Positive Acting Regulatory Genes, xy1R and xy1S, from the Pseudomonas putida HS1 TOL Plasmid PDK1

Description: TOL plasmids of Pseudomonas putida encode enzymes for the degradation of toluene and related aromatics. These genes are organized into two operons regulated by the Xy1R and Xy1S transcriptional activators. Previous analysis of the TOL pDK1 catechol-2,3-dioxygenase gene (xy1E) and a comparison of this gene to xy1E from the related TOL plasmid pWW0, revealed the existance of a substantial level of sequence homology (82%).
Date: May 1992
Creator: Chang, Teh-Tsai

Mechanisms of Cyanide Assimilation in Pseudomonas fluorescens NCIMB 11764

Description: Pseudomonas fluorescens NCIMB 11764 was capable of utilizing cyanide as a sole nitrogen source for growth. Cyanate (OCN") and S-cyanoalanine could also serve as nitrogenous substrates, but do not appear to play a role as intermediates in cyanide metabolism. Growth of this strain on cyanate as the sole nitrogen source led to the induction of an enzyme characterized as a cyanase (EC 3.5.5.3) based on its stoichiometric conversion of cyanate to ammonia, and dependence on bicarbonate for maximal activity. However, since cyanase activity was not elevated in cyanide-grown cells it was concluded that it serves no role in cyanide metabolism. Related studies aimed at examining a possible role for S-cyanoalanine as a cyanide-assimilation intermediate showed that while this compound also serves as a nitrogen source, it also is not important in cyanide metabolism. Studies focused on the utilization of free cyanide as a growth substrate led to the development of a fed-batch cultivation procedure greatly facilitating further experimentation aimed at the identification of cyanide metabolites. In addition to CO_2 and NH_3 as described earlier, two additional metabolites including formamide and formate were detected by using nC-NMR, HPLC, radioisotrapping methods and other analytical means. The formation of metabolites was shown to be induced after growth on cyanide with the relative product yields dependent on the availability of oxygen. These findings support earlier work in which an oxygen-dependent mechanism was proposed for the formation of C02 and NH3. However, at least two additional oxygen-independent pathways of cyanide conversion can be elaborated by this organism. One of these involves conversion to formate and ammonia while the other leads to the formation of formamide, which is not further degraded. Thus, growth on cyanide appears to occur by several mechanisms of chemical transformation presumably serving both detoxification and nutritional roles. Since two of these mechanisms ...
Date: August 1992
Creator: Nagappan, Olagappan

Molecular Phylogeny and Evolution of the American Woodrats, Genus Neotoma (Muridae)

Description: The evolutionary relationships of woodrats (Neotoma) were elulcidated through phylogenetic analyses of mitochondrial DNA restriction site and allozyme data. DNA samples from eleven nominal species from the genus Neotoma and two outgroup taxa, Ototylomys phyttotis and Xenomys nelsoni, were cleaved using a suite of 17 Type II restriction endonucleases. Mitochondrial DNA restriction profiles were visualized following electrophoresis of restriction digests via methods of Southern transfer and hybridization with 32P- and digoxigenin-labeled mtDNA probes. Restriction mapping resulted in the identification of 37 unique mtDNA haplotypes among the woodrat taxa examined. Proteins representing 24 presumptive structural gene loci were examined through starch gel electrophoresis. Binary-coded allozyme data and allozyme frequency data were analyzed using PAUP and FREQPARS, respectively. Phylogenetic analyses of the mtDNA restriction site data incorporated three different character type assumptions: unordered binary characters, Dollo characters, and differentially weighted unordered characters employing the STEPMATRIX option of PAUP. Proposed phylogenies for Neotoma are based on majority-rule consensus trees produced using bootstrap procedures. Phylogenetic analyses of the woodrat data sets revealed a distinct dichotomy among populations of white-throated woodrats (N. albigula) suggesting the presence of cryptic species within that taxon. MtDNA and allozyme data support the specific status of N. devia as distinct from N. lepida, and additionally reveal the presence of a third cryptic species referable to N. intermedia among the desert woodrats. Phylogenetic analyses of the genetic data also suggest subgeneric status for the desert woodrats, which is in agreement with evidence from morphology. The genetic data revealed a sister group relationship between N. stephensi and samples of N. mexicana, suggesting the placement of N. stephensi into the N. mexicana species-group. Neotoma fuscipes and N. cinerea formed a monophyletic lineage basal to the remaining members of the subgenus Neotoma which supports the assignment of N. fuscipes to the subgenus Teonoma with ...
Date: August 1992
Creator: Planz, John Valentine

Naloxone Potentiation of Epinephrine Induced Vasoconstriction in Canine Skeletal Muscle Arteries

Description: Naloxone (NX) potentiated epinephrine (EPI) induced submaximal vasoconstriction in canine renal and skeletal muscle arterial segments, yet had no vasoconstrictor action alone. Developed tension generated in-vitro by 4 x 1mm. O.D. rings from 1st degree branches of canine femoral arteries was expressed as % of KCI induced maximum response. NX (10^-5 M) potentiated EPI induced submaximal contractions (34.2%) significantly more than contractions induced by norepinephrine, phenylephrine, lofexidine, ADH, KCI and serotonin (13.8,13.4,4.7,13.5,14.4 and 11.4% respectively). The NX response was unaffected by beta-adrenergic blockade and NX did not reverse an isoproterenol mediated vasodilation. Alphaadrenergic blockade with phentolamine completely eliminated EPI plus NX induced vasoconstriction. After washout, vessels exposed to EPI plus NX relaxed by 50% significantly faster than vessels exposed to EPI alone (18.5 and 27.9 min respectively). EPI induced vasoconstrictions were potentiated by 10^-5 M corticosterone (49.0%) which inhibits extraneuronal catecholamine uptake, but not by 10^-7 M desipramine (1.1%) which inhibits neuronal uptake. EPI induced vasoconstrictions were also potentiated by 10^-4 M pyrogallol (33.0%) which inhibits catechol-o-methyl transferase activity, but not by 10^-5 M pargyline (-1.1%) which inhibits monoamine oxidase activity. The NX effect was endothelium independent. The dose-response of various opioid receptor agonists and antagonists were compared to the NX response. A specific opioid receptor subclass could not be identified as the mediator of the NX effect. The ED_50s for NX (3.7x^-6 M) and (+)NX (8.1x^-7M) indicated a significant stereoselectivity for the (+)enantiomer. A variety of sigma receptor ligands, steroids and steroid metabolites were tested for the ability to augment EPI vasoconstrictions. Several of the opioid, sigma and steroid ligands, all with polycyclic structures, induced responses similarto those of NX. NX exerted its effect independent of traditional opiate receptors and may have influenced the cellular uptake or degradation of EPI. Endogenous compounds with sigma or steroid activity may modulate ...
Date: August 1992
Creator: Stoll, Scott Thomas

Cardiorespiratory Responses to Graded Levels of Lower-body Positive Pressure During Dynamic Exercise in Man

Description: Cardiorespiratory responses to incremental dynamic exercise were assessed across four different levels of lower-body positive pressure (LBPP) and, as a separate study, during constant load (i.e constant work rate) exercise below and above each subject's ventilatory threshold (VT), both with and without 45 torr of LBPP.
Date: December 1992
Creator: Williamson, Jon W. (Jon Whitney)

The Eosinophil Response in Mice Infected with Trichinella spiralis or Trichinella pseudospiralis as Indicated by Phospholipase B Activity

Description: The host eosinophil response was compared in mice infected with either T. spiralis or T. pseudospiralis by determination of levels of splenic and intestinal phospholipase B, a marker enzyme for eosinophils. Primary infection of naive mice and challenge infection of homologously sensitized mice with T. pseudospiralis resulted in significantly lower tissue phospholipase B activities than infection with T. spiralis. Mice homologously challenged with T. pseudospiralis did exhibit an anamnestic eosinophil response compared to mice given a primary T. pseudospiralis infection. This anamnestic response, however, was significantly lower than the eosinophil response seen in sensitized mice given a homologous T. spiralis challenge. Mice sensitized to T. spiralis or T. pseudospiralis and heterologous challenge demonstrated an elevated eosinophil response compared to mice given a primary infection with either parasite. The heterologous challenge response, however, was not as intense as found for sensitized mice given a homologous challenge.
Date: December 1992
Creator: Hsu, Shing-Chien

Nucleotide Sequence Determination, Subcloning, Expression and Characterization of the xy1LT Region of the Pseudomonas putida TOL Plasmid pDK1

Description: The complete nucleotide sequence of the region encoding the DHCDH function of the pDK1 lower operon was determined. DNA analysis has shown the presence of two open reading frames, one gene consisting of 777 nucleotides encoding a polypeptide of 27.85 kDa and another gene of 303 nucleotides encoding a polypeptide of 11.13 kDa. The results of enzymatic expression studies suggest that DHCDH activity is associated only with xy1L. However although the addition of xy1T cell-free extracts to xy1L cell-free extracts does not produce an increase in DHCDH activity, subclones carrying both xy1L and xy1T exhibit 300- 400% more DHCDH activity than subclones carrying only xy1L.
Date: December 1992
Creator: Baker, Ronald F. (Ronald Fredrick)

Prediction of Post Mortem Interval from Degradation of Endogenous Nucleotides in Human Subjects

Description: High Performance Liguid Chromatography was used to measure degradation of nucleotides in human cadavers for the purpose of prediction of post mortem interval. Endogenous nucleotides were extracted from integumentary tissue of six(6) human cadavers using six percent(6%) tricholoacetic acid. Linear regression statistical techniques were used to determine linearity of degradation of various nucleotide pools.
Date: April 1993
Creator: Williams, John Burgess

Interactions between Carotid and Cardiopulmonary Baroreceptor Populations during Dynamic Exercise in Man

Description: During dynamic exercise the arterial baroreflexes have been thought to reset to the prevailing level of systemic pressure in order to modulate transient changes in blood pressure with the same sensitivity (gain) as at rest. To test this hypothesis, cardiovascular responses to dynamic exercise and carotid baroreflex responses to graded neck suction and neck pressure (NS/NP) were examined in seven men of moderate fitness (V02 = 41.4±3.6 ml O2*kg^-1*min^-1) during two levels (20% and 40% of peak oxygen uptake) of steady-state exercise. In addition, deactivation of cardiopulmonary baroreceptors has been thought to increase carotid baroreflex responsiveness in the quiescent state in man.
Date: May 1993
Creator: Potts, Jeffrey Thomas

Isolation and Characterization of the Operon Containing Aspartate Transcarbamoylase and Dihydroorotase from Pseudomonas aeruginosa

Description: The Pseudomonas aeruginosa ATCase was cloned and sequenced to determine the correct size, subunit composition and architecture of this pivotal enzyme in pyrimidine biosynthesis. During the course of this work, it was determined that the ATCase of Pseudomonas was not 360,000 Da but rather present in a complex of 484,000 Da consisting of two different polypeptides (36,000 Da and 44,000 Da) with an architecture similar to that of E. coli ATCase, 2(C3):3(r2). However, there was no regulatory polypeptide found in the Pseudomonas ATCase.
Date: May 1993
Creator: Vickrey, John F. (John Fredrick), 1959-

Nesting Ecology and Reproductive Correlates in the Desert-nesting Gray Gull Larus Modestus

Description: General objectives of my study were to describe the reproductive ecology of gray gulls in the large Lealtad colony, with emphasis on demographic parameters and physiological adaptations of eggs and chicks, which would complete some original objectives established in the early 1980's by Guerra and Fitzpatrick. Specifically, my study focused on describing, then comparing with other desert and non-desert nesting larids, interactive effects of ambient physical conditions and nest-site predation on eggs and chicks.
Date: May 1993
Creator: Aguilar Pulido, Roberto E. (Roberto Eric)