A study of the biology of Tropisternus lateralis, a hydrophilid beetle, was conducted during the flood period of a single playa on the Southern High Plains of Texas from early June 1995 through early September 1995. Mechanism of colonization, tolerance/avoidance to drought, larval density, and secondary production were analyzed. T. lateralis colonized playas from surrounding aquatic habitats and avoided drought through aerial dispersion. Once in the playa, larval density increased over time. Secondary production was 1.31 g/m2/.25 yr. In addition, aquatic Coleoptera diversity was studied in seven playas on the Southern High Plains of Texas. A total of twenty three species were identified from the study region. Nine species not reported in playa literature were identified.

Immunological and ultrastructural changes in mononuclear cells exposed to Mycobacterium avium serovar-specific glycopeptidolipid (GPL) and the chemically derived R-lipid (lipopeptide fragment) were examined.

Raptor rehabilitation has become commonplace globally, yet studies on the survival and adaptation of great horned owls (Bubo virginianus) after release has been neglected to an appreciable extent. The primary objective of this study is to provide quantitative data on the success of rehabilitated and released great horned owls in the North Texas region. Owls (N=12) were rehabilitated and released onto the Ray Roberts Greenbelt Corridor in Denton County, Texas, and monitored using radio telemetry to evaluate home range (November 2002 - February 2005). With approximately 75% of the birds released for this study surviving until transmitter battery failure, it is believed that the rehabilitation process was successful for these birds.

Life history allocation of energy to growth and reproduction was determined for female gizzard shad. Absolute caloric energy allocated to eggs increased with age. The relative amount of production energy directed to reproduction increased with age up to 5 years and then decreased. Seasonal variation in lipids was studied. Quantitative changes occurred in ovarian lipids during the reproductive cycle. Carcass lipids varied seasonally. Age of sexual maturity was 3 years. Delayed maturity is attributed to high allocation of energy to growth enabling shad to outgrow intense competition and predation in pre-reproductive ages; there is little competition and predation in reproductive shad. Growth rates and condition factors indicated constant availability of food seasonally.

This thesis describes the development of an optimized method for culturing dissociated, monolayer neuronal networks from murine frontal cortex and midbrain. It is presented as a guidebook for use by cell culture specialists and laboratory personnel who require updated and complete procedures for use with microelectrode array (MEA) recording technology. Specific cell culture protocols, contamination prevention and control, as well common problems encountered within the cell culture facility, are discussed. This volume offers value and utility to the rapidly expanding fields of MEA recording and neuronal cell culture. Due to increasing interest in determining the mechanisms underlying Parkinson's disease, the newly developed procedures for mesencephalon isolation and culture on MEAs are an important research contribution.

To examine if a rhythm can be entrained in either heart rate or oxygen consumption in late stage embryos (days 17-19.5) with light as a zeitgeber, chicken embryos were incubated in complete darkness (D:D) and 12:12 light:dark cycle (L:D). Light had no impact on oxygen consumption (390 µL O2∙min-1∙egg-1) but increased heart rate for non-internally pipped embryos (260 to 270 beats∙min-1 during light cycle). Oxygen consumption increased independent of pipping while heart rate increased (255 to 265 beats∙min-1) in D:D embryos due to pipping. A light-induced rhythm or effect occurred in heart rate but not oxygen consumption, suggesting heart rate and oxygen consumption may be uncoupled.

Spontaneously active frontal cortex and spinal cord networks grown on microelectrode arrays were used to study effects of D-methylphenidate. These central nervous system tissues have relatively low concentrations of dopaminergic and noradrenergic neurons compared to the richly populated loci, yet exhibit similar neurophysiological responses to methylphenidate. The spontaneous spike activity of both tissues was inhibited in a concentration-dependent manner by serial additions of 1-500 µM methylphenidate. Methylphenidate is non-toxic as spike inhibition was recovered following washes. The average concentrations for 50% spike rate inhibition (IC50 ± SD) were 118 ± 52 (n= 6) and 57 ± 43 (n = 11) for frontal cortex and spinal cord networks, respectively. A 3 hour exposure of a network to 1 mM methylphenidate was nontoxic. The effective concentrations described in this study are within the therapeutic dosage range. Therefore, the platform may be used for further investigations of drug mechanisms.

The role of individual amino acid residues of the enzyme dienelactone hydrolase was investigated. Using the polymerase chain reaction (PCR), a 1.9 kbp clcD fragment was amplified and subcloned yielding a 821 bp BamHI to EcoRI clcD subclone in the plasmid pUC19. Site-specific mutants of dienelactone hydrolase were created using mismatched oligonucleotides to prime DNA synthesis. Specifically modified proteins from mutated clcD genes (Arg 81 to alanine, Tyr 85 to phenylalanine and Arg 206 to alanine), were encoded by the mutant clones. Enzyme assays showed that dienelactone hydrolase activity of the mutants Arg 81 and Arg 206 was totally abolished. The DLHase enzyme activity of mutant Tyr 85 is greatly decreased by approximately two thirds.

Six reagent waters, eleven diets, and two culture sizes were evaluated for culturing C. dubia. Different filtration techniques were used to prepare the reagent waters. The eleven diets were comprised of two algae augmented with eight supplements. Reproduction and growth were assessed to discern differences among C. dubia raised in mass cultures and cultured in individual cups, during which, bacterial population densities, lipid, protein, and carbohydrate concentrations of the diets were measured. Results showed that a glass-distilled, carbon filtered, deionized reagent water and a Selenastrum capricornutum- Cerophyl® diet were optimum for culturing. Mass culturing supported the highest reproduction and growth, while no correlation was found between nutritional measurements and production.

After dendrite transection, two primary injury current pathways may acount for cell death: (1) the lesion current at the site of injury and (2) the voltage sensitive calcium channels along the dendrite. Lesions were made with a laser microbeam in mouse spinal monolayer cell cultures. Polylysine was tried as a positively charged "molecular bandage" to block the lesion current. The calcium channel blockers, verapamil and nifedipine, were used to reduce the calcium channel current. Control toxicity curves were obtained for all three compounds. The results show that neither verapamil, nifedipine, nor polylysine (MW: 3,300) protect nerve cells after dendrite amputation 100 ptm from the soma. The data also indicate that these compounds do not slow the process of cell death after such physical trauma.

Mean salinity concentrations in the four reservoirs (Moss, Ray Hubbard, Texoma and Possum Kingdom) ranged from 0.2 ppt in Moss Lake to 2.01 ppt in Possum Kingdom Lake. Reservoir sodium and chloride concentrations were hypotonic to hypertonic to plasma levels in all species. Interspecific differences were seen in ionic concentrations within each reservoir. Total osmotic and sodium concentrations in carp, Cyprinus carpio, were correlated to their concentrations in the reservoirs. No such relationship was noted for chloride, potassium and calcium. A laboratory study indicated that fish collection by electroshock did not bias plasma ion concentrations. Exposures to wide variations in ionic concentrations did not appear to induce stress in the species studied.

Seven species of planktonic microcrustacea were identified from the North Texas State University Golf Course Pond. Total adult microcrustacean community density, biomass and biocontent, and seasonal cycles of each species were compared with northern populations. Species diversity and evenness indices were highest in summer and lowest in spring. Variations in microcrustacean density showed a positive correlation with density of phytoplankton. Temperature had a direct effect on metabolic rates of two species of copepods. Metabolic rates of pond species were lower at common temperatures than those of northern populations. An estimate of annual energy flow through the pond ecosystem showed cladocerans contributed the greater percentage of total energy to the next trophic level.

Bacteria classified in the family Methylomonadaceae must derive their carbon from one-carbon compounds. They are characterized by the possession of internal membranes of two types. Type I membranes are layered and fill the middle of the cells while type II membranes form concentric layers around the periphery of the cells. Also, there are two metabolic pathways by which the methylobacteria assimilate one-carbon compounds. Further evidence of this dichotomy was sought by DNA-DNA saturation hybridization of DNAs from both types of methylobacteria. Very low DNA-DNA homology was seen between types I and II or within the types. It was not possible, therefore, to correlate the degree of genetic relatedness with either the nature of the internal membranes or the pathway of carbon assimilation.

Electrophoretic variation in 12 proteins encoded by 16 loci was analyzed to compare the genetic relationships of 18 natural populations representing two species of stoneroller minnows, Campostoma anomalum and C. oligolepis. Ten of the loci were monomorphic and fixed for the same allele in all populations of both species. One locus, Mdh-2, was found to separate both species. Mean heterozygosity for both species was 0.072. Estimates of levels of inbreeding indicated this phenomenon is operating in C. anomalum to structure the populations genetically. Mean genic identity (I) between the two species was high, 0.887, indicating the taxa are closely related. Nevertheless, data accumulated point to the conclusion that the two species maintain electrophoretic variation in 12 proteins encoded by 16 loci was analyzed to compare the genetic relationships of 18 natural populations representing two species of stoneroller minnows, Campostoma anomalum and C. oligolepis. Ten of the loci were monomorphic and fixed for the same allele in all populations of both species. One locus, Mdh-2, was found to separate both species. Mean heterozygosity for both species was 0.072. Estimates of levels of inbreeding indicated this phenomenon is operating in C. anomalum to structure the populations genetically. Mean genic identity (I) between …

Drumming is first described for five North American stonefly species, Acroneuria evoluta, Doroneuria baumanni, Isoperla namata, Chernokrilus misnomus, and Pictetiella expansa. Signals of Acroneuria lycorias, Phasganophora capitata and Isoperla signata are further described. Drumming was not recorded from Amhinemura delosa. Signals of A. evoluta are the most complex yet recorded in Plecoptera. Doroneuria baumanni, P. expanse, C. misnomus and P. capitata have 2-way exchanges. Male D. baumanni produce two prolonged beats by rubbing the hammer on the substratum; male-female signals are non-overlapping in the first two species and overlapping in the latter two. Female P. capitata answered with an unusually long sequence of beats. Two male Isoperla species produced monophasic calls without female answers. Female A. lycorias answered taped male signals with monophasic signals like all observed females.

N^6,O^2' -Dibutyryl cyclic adenosine 3',5'-monophosphate (Bt_2cAMP) was investigated for its effects on various tissue culture cells infected with temperature-sensitive (ts) mutant, LA31 and Bratislava 77 (B77), a wild-type Rous sarcoma virus. Specifically, known parameters of transformation were investigated and a possible site of action has been tenably proposed. The drug Bt_2cAMP was found to have little effect on the transformation related properties of primary chick embryo fibroblasts (CEF) infected with either virus or normal rat kidney fibroblasts (NRK) infected with the wild-type B77-RSV. However, significant inhibition of the transforming properties in NRK infected with the ts mutant LA31 (LA31-NRK) were reported at the permissive temperature 33 degrees centigrade (33 C).

The purpose of this study was to focus attention on the physiology of the bdellovibrio-host interaction and to determine the metabolic requirements for this reaction. Since bdellovibrio is an aerobic organism, direct measurements of respiration, turbidity, and viable cell counts are reliable indications of the metabolic activity of the cells. It was determined that the metabolic requirements for the parasitic interaction are constituents from either metabolically active host cells or cells which are capable of at least some metabolic activity. The nutritional requirements of host-independent bdellovibrios suspended in buffer are not met by the presence or absence of viable or nonviable Enterobacter aegnes. Unlike the HD bdellovibrios, the HI bdellovibrios lack the ability to make economical use of their self-digesting processes.

Analysis of larval fish collected at four stations in a 330-ha cooling reservoir indicated Dorosoma spp. were most numerous at all stations, followed by Lepomis spp. and Percina Macrolepida. Largest numbers and greatest diversity of larval fish were found at the station least affected by thermal effluent; the mid-lake station provided the smallest numbers and least diversity. The two warmwater stations were intermediate, with similar numbers and diversity. Diversity and abundance of zooplankton between stations were similar to those of fish. The most abundant zooplankter (Bosmina) was generally selected against by Dorosoma, Lepomis and Micropterus spp. larvae except when the larvae were quite small ((10mm). Cyclopoid copepods were most often selected by all larvae.

Chick embryo fibroblasts and rat kidney cells infected in vitro with Rous Sarcoma viruses were treated with 3-deazaguanine (3-DG). The findings revealed that 3-DG inhibited virus-induced cellular transformation. Degree of inhibition is dependent on concentration and frequency of media change. 3-DG at the concentrations tested will not reverse transformed cells to the untransformed state and does not have marked effect on replication of viruses. Upon removal of 3-DG, its effect was shown to be reversible. Cell growth was generally retarded in medium containing 3-DG. When xanthosine and inosine were added to the medium, cell growth was unaffected, but it increased in guanosine.

A procedure for the isolation of malic enzyme from muscle tissue of the roundworm Ascaris suum is described. The fractionation method yields relatively large quantities of the enzyme,with a specific activity of fifteen moles of malate converted to pyruvate and carbon dioxide per min per mg at 25º. Homogeneity was established with analytical ultracentrifugation, zone electrophoresis, isoelectric focusing, and rechromatography. The molecular weight of the enzyme was 250,000, and it is dissociated under several conditions into four identical monomers of 64,000 daltons. The enzyme exists as a single electrophoretic form and prefers manganous and NAD over other cations and NADP. Ammonium sulfate competes with manganous for the active site and titration with DTNB yields eight thiol groups per mole. Titration of the first four thiol groups is accompanied by a complete loss in enzyme activity. Equilibrium dialysis, product inhibition, and initial velocity studies suggest a rapid-equilibrium random sequential mechanism for the Ascaris suum malic enzyme. The presence of 1.3 binding sites per subunits was determined for L-ma late. Antisera prepared against A. suum malic enzyme reacted to a small extent with the NAD malic enzymes from two free-living nematodes, Panarellus redivivus and Turbatrix aceti. A correlation coefficient of 0.911 was …

An examination of the Desmodium canescens complex (D. canescens; D. tweedyi; D. illinoense) has resulted in the delimitation of a previously unreported alliance between D. canescens and D. tweedyi. The following points support this view: (a) morphological data taken from herbarium and garden specimens indicate that for many characters, the mean values of D. canescens and D. tweedy are not significantly different (b) breeding experiments have shown that artificial interspecific hybridization is possible between D. canescens and D. tweedyi (c) cytological studies have shown that D. canescens and D. tweedyi have a base number of x = 11, while D. illinoense has a base number of x = 10. A new combination is suggested: Desmodium canescens var. tweedyi (Britt.) Williams.

Proteomic techniques were used to evaluate the protein profile of the earthworm, (Lumbricus terrestris), following a bacterial challenge. One control group received no injection; a second control group received injections of phosphate buffer solution (PBS). The experimental group received injections of PBS containing (Aeromonas hydrophila). After incubation for 12 hours at 20°C, coelomic fluid was collected from each group for analysis by 2-D electrophoresis. There were significant differences in spot appearance and density between control and experimental groups. Sixteen spots showed a two-fold increase in density and 63 showed at least a two-fold decrease in density between samples from control and bacteria-challenged earthworms, respectively, suggesting up- and down-modulation of proteins potentially involved in the earthworm's response to bacterial challenge.

The chicken ductus arteriosi (DA) are two embryonic blood vessels that shunt blood away from the non-ventilated lungs and towards the body and chorioallantoic membrane. I show that prostaglandins have a diminished role in maintaining chicken DA patency and nitric oxide inhibits oxygen induced contraction of the day 19 proximal DA in a time dependent manner. The pathways governing oxygen induced contraction in the chicken DA are similar to those found in mammals and include contributions from ROS, Kv channels, L-type Ca2+ channels, and the Rho kinase pathway. Longer exposure to high oxygen generates increased oxygen induced constriction of the day 19 DA that may be mediated through the Rho kinase pathway.

This study focused on the enzyme, aspartate transcarbamoylase (ATCase) from A. hydrophila, a Gram-negative bacterium found in fresh water. The molecular mass of the ATCase holoenzyme from A. hydrophila is 310 kDa. The enzyme is likely composed of 6 catalytic polypeptides of 34 kDa each and 6 regulatory polypeptides of 17 kDa each. The velocity-substrate curve for A. hydrophila ATCase is sigmoidal for both aspartate and carbamoylphosphate. The Km for aspartate was the highest to date for an enteric bacterium at 97.18 mM. The Km for carbamoylphosphate was 1.18 mM. When heated to 60 ºC, the specific activity of the enzyme dropped by more than 50 %. When heated to 100 ºC, the enzyme showed no activity. The enzyme's activity was inhibited by ATP, CTP or UTP.