Digital Commons Network^™

Denervation Supersensitivity Of The Rat Vas Deferens: A Role For Protein Kinase C, Sonny T. Abraham

Electronic Theses and Dissertations

A role for protein kinase C (PKC) in the denervation-induced supersensitivity of the rat vas deferens was investigated. Chronic, surgical denervation of the rat vas deferens (up to 8 days) resulted in tissues that produced enhanced contractile responses to norepinephrine (NE) in isolated organ baths. Single challenges of NE (10 $\mu$M) produced 0.6 $\pm$ 0.1 g of maximal tension in the control vas whereas in the paired, denervated tissue 2.2 $\pm$ 0.3 g of tension was recorded (n = 6). Cumulative concentration-effect curves to NE produced in the denervated vas deferens were shifted 18-fold to the left of the control …

Differential Role Of The Endothelium In Regulating Microvascular Blood Flow, Tao Tang

Electronic Theses and Dissertations

The vascular endothelial cell (EC) plays an important role in regulating vascular tone and local blood flow by sensing chemical and mechanical stimuli on the vascular wall and releasing a host of vasoactive substances upon activations of endogenous or exogenous vasoactive substances. The central hypothesis is that local control of blood now and autoregulatory behavior in the microcirculation is distinctive at different levels of the vasculature and is dependent on the cellular activities of the EC and its interaction with the local environment. The in vivo as well as the ex vivo, flow-controlled preparations of the hamster cheek pouch were …

Characterization Of The Vasoactivity Of Tachykinins In Isolated Rat Kidney: Functional Studies And In Vitro Receptor Autoradiography, Yuejin Chen

Electronic Theses and Dissertations

Although tachykinins have potent vascular actions, their effect on renal resistance blood vessels is currently unknown. The vasoactive properties of tachykinins and related analogs were assessed in isolated perfused rat kidney. At a basal perfusion pressure (PP) of 75 $\pm$ 6 mm Hg (n = 5), bolus injections of substance P (SP) had no significant vasoactive effect. Following a sustained increase in baseline PP (134 $\pm$ 10 mm Hg) produced by phenylephrine (1 $\mu$M), SP evoked a dose-dependent increase in PP. The largest dose of SP increased PP by 60 $\pm$ 5 mm Hg. The vasoconstrictor response to SP was …

A Molecular Basis For Erythromycin Sensitivity And Resistance In Escherichia Coli, Harold S. Chittum

Electronic Theses and Dissertations

The effect of erythromycin on the 50S ribosomal subunit during cell growth has been extensively investigated. Sucrose density gradient analysis of ribosomes formed in the presence and absence of the drug revealed a 50S specific assembly defect is partially responsible for erythromycin's inhibitory effects on wild type cells. Examination of two erythromycin-resistant mutants of E. coli (N281 and N282) revealed that mutant N281 (L22 mutant) but not N282 (L4 mutant) was assembly defective in the presence of the drug, although only at much higher drug concentrations (300 ug/ml vs. 75 ug/ml for wild type cells). The altered genes from each …

A Characterization Of Extractable, Hydroxylated Fatty Acid Bearing Components In Legionella Pneumophila, Jonathan R. Lane

Electronic Theses and Dissertations

Extraction of the lipids of Legionella pneumophila yields phases unlike those produced from other Gram-negative bacteria. A viscous interface forms between the aqueous (wash) and organic phases. More than half of the hydroxylated fatty acids were found distributed between the aqueous phase and the interfacial material, fractions in which such constituents have not been reported in other Gram-negative species. It was further observed that after the material from the aqueous/interfacial phase was dissolved in methanol or chloroform/methanol (2:1 (V/V)), the addition of acetone would create a white, flocculent precipitate. Analyses showed that the supernatant contained fatty acids that were nonhydroxylated …

Nitric Oxide Production: A Mechanism For Inhibition Of Chlamydia Trachomatis Replication, Bojun Chen

Electronic Theses and Dissertations

Chlamydia trachomatis (CT) replicates in macrophages, but is inhibited by IFN-$\gamma$ or LPS. IFN-$\gamma$ and/or LPS induced nitrite production in mouse peritoneal macrophages, macrophage cell lines (RAW264.7 and J774A.1) and McCoy cells. Kinetic studies indicated that peak production occurred 48 hours post-treatment. CT infection itself was insufficient to induce nitrite production, but resulted in enhancement of nitrite production in IFN-$\gamma$-treated cells. Treatment with IFN-$\gamma$ or LPS resulted in significant inhibition of CT replication in these cells. Strong correlation between nitrite production and inhibition of CT replication was observed in RAW264.7 and J774A.1 cells (correlation coefficients: $-$0.93 and $-$0.94, p $<$ 0.001). N$\sp{\rm g}$- monomethyl-L-arginine (L-NMMA) specifically inhibited nitrite production and partially reversed inhibition of CT replication in macrophage cell lines. NOS mRNA was measured in RAW264.7 cells by Northern blot and Dot blot hybridization. Strong correlation between NOS mRNA expression and inhibition of CT replication (correlation coefficient: $-$0.97, p $<$ 0.05) was observed. Anti-TNF-$\alpha$ antibody completely neutralized the biological activity of TNF-$\alpha$ secreted by LPS-treated RAW264.7 cells, yet the antibody neither reduced nitrite production nor restored CT replication. Combination of the antibody and L-NMMA significantly enhanced restoration of CT replication. In peritoneal macrophages, inhibition of CT replication induced by IFN-$\gamma$ was partially restored by L-NMMA or anti-TNF-$\alpha$ antibody. In McCoy cells, inhibition of CT replication induced by IFN-$\gamma$ and LPS was not significantly restored by L-NMMA. Great restoration of CT replication by 1 mM L-NMMA was observed in LPS-treated J774A.1 cells (31%), but not in IFN-$\gamma$-treated cells (5%). Our data indicate that (1) NO production is one of the mechanisms for inhibition of CT replication in IFN-$\gamma$-activated peritoneal macrophages and RAW264.7 cells; (2) NO plays a significant role in CT inhibition in LPS-treated macrophage cell lines, but not peritoneal macrophages; (3) TNF-$\alpha$ may be associated with inhibition, but the mechanism(s) may not involve NO production; (4) NO production may not be the mechanism for CT inhibition in McCoy cells treated with IFN-$\gamma$ and LPS.

Co-Sensitization Of Dopamine And Serotonin Receptors Occurs In The Absence Of A Change In The Dopamine D1 Receptor Complex After A Neonatal 6-Ohda Lesion, Li Gong

Electronic Theses and Dissertations

To test whether SKF 38393 could ontogenetically sensitize dopamine (DA) D$\sb1$ receptors and whether this sensitization would be associated with biochemical changes, intact and neonatal 6-hydroxydopamine (6-OHDA)-lesioned rats (200 $\mu$g i.c.v.) were treated daily from birth with SKF 38393 (3.0 mg/kg i.p. x 28 days) or its vehicle. In DA D$\sb1$ neonatally sensitized 6-OHDA rats, enhanced locomotor responses were observed with the first SKF 38393 challenge dose (3.0 mg/kg i.p.) at 6 weeks. This response increased further with weekly SKF 38393 treatments. Enhanced stereotyped behaviors were seen in both lesioned and sensitized rats at 8 weeks. There was no change …

Unusual Structure Of A Human Middle Repetitive Dna, Duminda D. Ratnasinghe

Electronic Theses and Dissertations

The L2Hs sequences are a polymorphic, interspersed, middle repetitive DNA family unique to human genomes. Genomic fingerprinting indicates that these DNAs vary from one individual to another and between tissues of the same individual. Sequence analysis reveals that they are AT-rich (76%) and contain many unusual sequence arrangements (palindromes, inverted and direct repeats). These sequence properties confer on the L2Hs elements the potential to fold into non-B-form structures, a characteristic of recombination hot spots. To test this hypothesis carbodiimide, osmium tetroxide and S$\sb1$ nuclease were used as single-strand specific probes to study a recombinant plasmid, pN6.4.39, containing a single L2Hs …

The Effect Of Trimethyltin On The Cholinergic System Of The Rat Hippocampus, Richard L. Cannon

Electronic Theses and Dissertations

Trimethyltin (TMT) is a neurotoxin occurring in the environment. Exposure to (TMT) is known to destroy specific neuronal components of the hippocampus in the rat and to cause clinical symptoms in exposed humans, including mnemonic deficits, that indicate hippocampal involvement. In addition to hippocampal cell loss TMT causes significant increases in cholinergic markers such as acetylcholinesterase (AChE) stain density and choline acetyltransferase (ChAT) activity in the hippocampus of rats. However, despite these observations the effect of TMT on hippocampal cholinergic system has not been investigated in detail. The purpose of the present study was to elucidate more fully the consequences …

Changes In Intracellular Chloride During Osmotic Stress And L-Alanine Uptake In Mouse Hepatocytes, Kening Wang

Electronic Theses and Dissertations

A stable intracellular ionic environment is necessary for hepatocytes to function normally. Thus, during hypotonic shock or L-alanine uptake, hepatocytes swell and then exhibit a regulatory volume decrease (RVD), which comprises an increase in K$\sp+$ conductance (G$\sb{\rm K}$), an increased K$\sp+$ efflux, and a hyperpolarization of transmembrane potential (V$\sb{\rm m}$). Since hepatocyte intracellular Cl$\sp-$ has been demonstrated to distribute passively with V$\sb{\rm m}$, this study is designed to test the hypothesis that the hypotonic shock- or L-alanine uptake-induced hyperpolarization of V$\sb{\rm m}$ might provide an electromotive force for the efflux of hepatocyte intracellular Cl$\sp-$, which in turn would contribute osmotically …

Molecular And Cellular Analysis Of Chlamydia Trachomatis: Persistence And Reactivation, Kimberly R. Tau

Electronic Theses and Dissertations

Chlamydia trachomatis (CT) is the most prevalent sexually-transmitted infection in the United States. It has been suggested that CT infections can become latent. This has not been substantiated. CT persistence was examined at the molecular and cellular level in vitro and in vivo. Penicillin treatment of CT in vitro results in abnormal inclusions and reduced recovery of infectious CT. Penicillin did not inhibit initial stages of infection, but did downregulate CT rRNA levels after 25 hours post-inoculation (p.i.). DNA amplification was employed to differentiate between a resolved infection and a persistent one. Utilizing a primer pair that amplified a 144 …

Development Of Mass-Rearing Techniques For Aphelinus Asychis Walker For Use In A Biological Control Program Against Russian Wheat Aphid, Robin Lynn Whitaker-Deerberg

Theses and Dissertations - UTB/UTPA

Mass-rearing techniques were developed for the parasitoid Aphelinus asychis Walker (Hymenoptera:Aphelinidae) for use in a biological control program against the Russian wheat aphid, Diuraphis noxia (Mordvilko) (Homoptera:Aphididae). Daily ovipositional activity, host-feeding behavior, effects of population density in the rearing cages and effects of cold storage on mummies were determined.

Daily ovipositional activity peaked at day 5 after emergence of the parasitoid. Ninety-two percent of mummies resulting from ovipositional activity were oviposited the first 13 days of the parasitoid’s life. Since mummification occurred 11 days after oviposition and emergence occurred 11 days after mummification, day 24 was determined to be the …

Characterization Of Two Temperature-Sensitive Mutants Of Escherichia Coli Exhibiting An Altered L22 Ribosomal Protein, Bonnie A. Burnette-Vick

Electronic Theses and Dissertations

Analysis of E. coli strains SK1047 and SK1048 have shown them to be temperature-sensitive, protein-synthesis deficient. An alteration in ribosomal protein L22 was detected in both strains using two dimensional gel electrophoresis. Protein L22 was purified from both strains by reversed phase high performance liquid chromatography and from two dimensional electrophoretic gels. Purified ribosomal protein L22 was labeled by reductive methylation and used in 23S RNA binding assays with and without ribosomal protein L4. At the permissive temperature, protein L22 from SK1047 bound less efficiently than the control while protein L22 from SK1048 bound as efficiently as the control. At …

Mouse Mast Cell Proteases: Induction, Molecular Cloning, And Characterization, Wei Chu

Electronic Theses and Dissertations

Tryptase, a mast cell-specific serine protease with trypsin-like specificity, has been identified in a mouse mast cell line (ABFTL-6) based on it's enzymatic activity, inhibition properties, and cross-reactivity to a human mast cell tryptase antibody. The effects of fibroblast-conditioned medium and sodium butyrate on ABFTL-6 mast cell differentiation and tryptase expression have been examined. ABFTL-6 mouse mast cells undergo phenotypic changes upon culturing in media supplemented with fibroblast-conditioned media at 50% or 1 mM sodium butyrate. The induced cells increased in size, had larger and more metachromatic cytoplasmic granules, and increased their total cellular protein about four-fold. Tryptase activity increased …

A Temperature-Sensitive Mutant Of Escherichia Coli Affected In The Alpha Subunit Of Rna Polymerase, Majid Mehrpouyan

Electronic Theses and Dissertations

A temperature-sensitive mutant of Escherichia coli affected in the alpha subunit of RNA polymerase has been investigated. Gene mapping and complementation experiments placed the mutation to temperature-sensitivity within the alpha operon at 72 min on the bacterial chromosome. The rate of RNA synthesis in vivo and the accumulation of ribosomal RNA were significantly reduced in the mutant at 44$\sp\circ$C. The thermostability at 44$\sp\circ$C of the purified holoenzyme from mutant cells was about 20% of that of the normal enzyme. Assays with T7 DNA as a template showed that the fraction of active enzyme competent for transcription was reduced as a …

Electrophysiology, Cell Calcium, And Mechanisms Of Hepatocyte Volume Regulation, Walid E. Khalbuss

Electronic Theses and Dissertations

The electrophysiologic technique (Reuss, L., Proc. Natl. Acad. Sci. USA 82:6014, 1985) was modified to measure changes in steady-state hepatocyte volume during osmotic stress. Hepatocytes in mouse liver slices were loaded with tetramethylammonium ion (TMA$\sp{+}$) during transient exposure of cells to nystatin. Intracellular TMA$\sp{+}$ activity (a$\sp{\rm i}\sb{\rm TMA}$) was measured with TMA$\sp{+}$-sensitive, double-barreled microelectrodes. Loading hepatocytes with TMA$\sp{+}$ did not change their membrane potential (V$\sb{\rm m}$), and under steady-state conditions a$\sp{\rm i}\sb{\rm TMA}$ remained constant over 4 min in single impalements. Hyperosmotic solutions (50, 100, & 150 mM sucrose added to media) and hyposmotic solutions (sucrose in media reduced by …

Variants And Polymorphisms Of Three Repetitive Dna Families In The Human Genome, Robert M. Roudabush

Electronic Theses and Dissertations

A novel 0.6 kb LINE family in human DNA, designated L2Hs, has been described (Musich and Dykes 1986). Studies employing clone N6.4, containing three 0.6 kb segments of this family, indicate that these sequences are interspersed and moderately repetitive. Two additional variant sequences of the L2Hs family, N6.1 and N6.3, have been identified. Restriction mapping of each cloned segment indicates similarities among N6.4, N6.3 and N6.1. When the cloned DNAs were cleaved with restriction enzymes and subjected to cross-hybridization, each cloned insert produced a pattern indicating that the sequences contained in N6.1 and N6.3 are represented in at least one …

Seasonal Distribution Of The Ichthyofauna Of Santa Ana National Wildlife Refuge, Alamo, Texas, Matthew Albert Ciomperlik

Theses and Dissertations - UTB/UTPA

This study investigated the small aquatic ecosystems of Santa Ana National Wildlife Refuge, located in a predominantly agricultural area of the Lower Rio Grande Valley, Texas. Fishes and water quality data were collected for 13 months to determine the status of fish populations and the factors which influence their distributions on the refuge.

Sixteen fish species were collected during the study, however, more than 20 species are known to inhabit the Rio Grande in the vicinity of the refuge. Water source played a major role in influencing fish species richness in these aquatic ecosystems. Diverted river water offered the best …

Quantitation Of Teratogenic Effects Of 5-Fluorouracil Administered To Mice In Vivo Or In Submerged Limb Culture, Theresa A. Sanders

Electronic Theses and Dissertations

This study demonstrates the use of submerged limb culture in teratologic testing. Pregnant mice were treated on day 11 of gestation (E11, plug date = E0) with 10, 20 or 40 mg of 5-fluorouracil (FU) per kg body weight. On E17, treated and untreated fetuses were examined for gross malformations and were fixed in 95% ethanol. Reduction of limb size and digital defects, including ectrodactyly (ED), syndactyly (SD), microdactyly and polydactyly were dose-dependent. In parallel studies, pregnant mice were treated on the morning of E11 and embryos were removed either 7h (E11) or 24h (E12) later for submerged limb culture. …

Adaptation Of Striped Bass To Sea Water Following Direct Transfer From Freshwater: Morphological, Biochemical, And Physiological Parameters, Judy A. King

Electronic Theses and Dissertations

There has been heightened interest in the biology of striped bass (Morone saxatilis) because of increased pollution in their native spawning grounds and because of their extensive use in landlocked sport fisheries. Their euryhalinity makes them an excellent species for osmoregulation studies. The objective of this research was to study the rate of adaptation of striped bass gills to sea water (3% salt) after direct transfer from freshwater using biochemical (ion transport enzyme levels), physiological (chloride efflux), and ultrastructural methods. Striped bass have specialized osmoregulatory cells located on the interlamellar and afferent surfaces of their gill filaments as shown by …

Isolation And Characterization Of Temperature-Sensitive Protein Synthesis Mutants Of Escherichia Coli By Directed Mutagenesis Of The Defective Bacteriophage Lambda Fus2, Kenton L. Lohman

Electronic Theses and Dissertations

Mutagenesis of the defective transducing bacteriophage lambda fus2 was used to isolate a collection of temperature-sensitive mutants of E. coli in the major ribosomal protein gene cluster. Four mutants were examined in detail. Two of the mutants were resistant to the ribosomal antibiotics neamine and spectinomycin. Another mutant was defective in 50S ribosomal subunit assembly at 42(DEGREES)C. The 30S subunit proteins S17 and S19 were changed in two different mutants. Each protein migrated as a more basic species in two-dimensional gels of ribosomal proteins. Ribosomes from each of the four mutants examined showed a temperature-dependent reduction in translational activity in …

Characterization Of Kpni Interspersed, Repetitive Dna Sequence Families And Their Association With The Nuclear Matrix, Joseph A. Chimera

Electronic Theses and Dissertations

The KpnI, 1.2 and 1.5 kb families of interspersed repetitive DNAs from the African green monkey genome were isolated and characterized. Each family contains three populations of segments based on their sequence lengths and susceptibility to cleavage by the restriction enzymes KpnI and RsaI. The first population contains the smallest segments which are susceptible to both KpnI and RsaI cleavage and have fragment lengths of 1.2 kb (1.2 kb family) and 1.5 kb (1.5 kb family) respectively. The members in this population are referred to as KpnI-sensitive segments. The second population contains longer segments (> 2 kb) which represent fusions …

Eye Movement Related Single-Unit Recording In The Nucleus Reticularis Tegmenti Pontis In The Alert Monkey, William Franklin Crandall Jr.

University of the Pacific Theses and Dissertations

Nucleus Reticularis Tegmenti Pontis (NRTP) is a structure which lies in the mid-brain ventral to the oculomotor complex. Anatomical studies strongly implicate it in oculomotor function as most of its inputs and outputs are to systems previously shown to be intimately involved in eye movement generation--superior colliculus (SC) and cerebellum. These reports show an integrative structure with inputs and outputs distributed in a way well suited for performing relay and feedback tasks. Physiological data in the alert, trained monkey has been gathered in an attempt to characterize these neurons and to classify them in functional terms. Three monkeys were trained …

Aspects Of The Oculomotor System Of Callinectes Sapidus, Antoinette Steinacker

University of the Pacific Theses and Dissertations

An isolated perfused preparation was developed for the study of several aspects of the oculomotor system of the blue crab, Callinectes sapidus. The system for eyestalk rotation was investigated on an extracellular level. Two antagonistic pairs of muscles under visual and statocyst control were found to be responsible for stabilization and rotation of the eyestalk. The primary sensory input to the muscles appears to be from the statocysts, with both static position sense and dynamic acceleration components influencing the motor response. Two sensory feedback systems from mechanoreceptive hairs were found which influence the response of the eye stalks to statocyst …