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Acute Effects of the Antibiotic Streptomycin on Neural Network Activity and Pharmacological Responses
The purpose of this study is to find out that if antibiotic streptomycin decreases neuronal network activity or affects the pharmacological responses. The experiments in this study were conducted via MEA (multi-electrode array) technology which records neuronal activity from devices that have multiple small electrodes, serve as neural interfaces connecting neurons to electronic circuitry. The result of this study shows that streptomycin lowered the spike production of neuronal network, and also, sensitization was seen when neuronal network pre-exposed to streptomycin.
The aquatic insect communities of Holbrook Creek and Cochetopa Creek in Colorado.
The first objective for this problem in lieu of thesis project was to gather, identify to the lowest practical taxonomic level and organize all available aquatic insects collected from high altitude Colorado aquatic systems during the summers of 1994, 1996, 1998, and 2002 for the University of North Texas Environmental Science Field Course (BIOL 5650). The curated collection will be housed in the Elm Fork Natural History Museum, located at the University of North Texas. The second objective was to provide a summary and discussion of the occurrence and distribution of the aquatic insects collected from Mt. Blanca in 1994, 1996, and 1998 and to create a taxa list of aquatic insects collected from Cochetopa Creek during the summer of 2002.
Aryl Hydrocarbon Hydroxylase and Sixteen Alpha Hydroxylase in Cultured Human Lymphocytes
Cultured human lymphocytes may be assayed for aryl hydrocarbon hydroxylase (AHH) in whole cell preparations. The optimum assay conditions are pH 8.5, and 1.5 mM Mg++. The reaction is linear with time and cell number, and is inhibited by CO. Estradiol may inhibit induction of AHH by 3-methylcholanthrene, but is a poor competitor for the enzyme. A Caucasian population was assayed for AHH activity. The distribution was lognormal; no difference was found in cultured cells from males and females or smokers and nonsmokers. Cells from relatives of lung cancer patients showed higher activity. An American Indian population showed no difference from the Caucasian population in enzyme level. No linkage was found between AHH and 16a-hydroxylase.
Aspartate Transcarbamoylase of Aeromonas Hydrophila
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.
Attenuation of Escherichia Coli Aspartate Transcarbamoylase Expressed in Pseudomonas Aeruginosa Mutant and Wild Type Strains
No apparent repression of pyr gene expression in Pseudomonas aeruginosa is observed upon addition of exogenous pyrimidines to the growth medium. Upon introduction of the subcloned Escherichia coli pyrBI genes for aspartate transcarbamoylase (ATCase) into a P. aeruginosa pyrB mutant strain, repression was observed in response to exogenously fed pyrimidine compounds. The results proved that it is possible to bring about changes in pyrimidine nucleotide pool levels and changes in transcriptional regulation of gene expression as a result. Thus, the lack of regulatory control in P. aeruginosa pyr gene expression is not due to an inability to take up and incorporate pyrimidine compounds into metabolic pools, or to an inability of the RNA polymerase to respond to regulatory sequences in the DNA but is probably due to a lack of specific regulatory signals in the promoter of the genes themselves.
Autonomic Nerve Activity and Cardiovascular Function in the Chicken Embryo (Gallus gallus)
The goal of this study was to build on the historic use of the avian model of development and also to further the knowledge of autonomic nervous system (ANS) regulation of cardiovascular function in vertebrates. Vasoactive drugs sodium nitroprusside, a vasodilator and phenylephrine, a vasoconstrictor were used to study the correlation of cardiovascular function relationship with nerve activity, both sympathetic and parasympathetic (vagal). Additionally, ANG II was used to assess its effects on vagal inhibition. The present study shows that pharmacologically-induced hypertension is associated with a fall in mSNA, indicating that the capacity for sympathetic autonomic cardiovascular regulation is established by late incubation however, late-stage embryonic chickens did not show a significant increase in mSNA during hypotension. The hypotensive response of the embryo was not accompanied by the expected inhibition of vagal discharge; however a slight but insignificant reduction in vagal discharge was noted. When vagal efferent output was isolated, a significant drop in vagal efferent activity was noted in response to hypotension. The present study showed late-stage embryonic chickens lack a vagal response to hypertension in both efferent and sensory limbs. In this study, vagal discharge was reduced from baseline levels in response to Ang II. Collectively, the present study indicates that the lack of a decreased heart rate, in response to increases in Pm caused by Ang II, is due to a central inhibitory action of Ang II on the vagus. Data from the present study suggests that although autonomic interaction with the cardiovascular system in present in late-stage chicken embryos, it is still underdeveloped and possesses a limited capacity.
Beta-adrenergic Blockade Via Atenolol and Its Effects on Blood Pressure, Heart Rate, and Renal Morphology in the Developing Chicken Gallus Gallus Domesticus
Chicken embryos were chronically exposed to the ?1- blocker atenolol during one of three stages: mesonephros (E7-E9), mesonephros-metanephros (E11-E13), or metanephros (E15-E17). Mesonephros group hearts were larger than all other groups (P < 0.01). Mesonephros and metanephros group kidneys were larger than all remaining groups (P < 0.0001). The mesonephros group nephron number was ~40% lower than control values (P = 0.002). Glomerular areas were 26% and 18% larger than the control group in the mesonephros and metanephros groups, respectively (P < 0.001). These data suggest an E7-E9 critical window of cardiovascular and renal development for atenolol. Acute atenolol exposure in E15 embryos showed an increase in mean arterial pressure with all but the highest dose. All doses significantly decreased heart rate.
Biochemical Genetics of Certain Species of the Blackbird Family Icteridae
Starch gel electrophoresis was used to compare 14 proteins encoded by 15 loci for seven species of the family Icteridae. A close genetic relationship among these species was classified into three groups. The Agelaiine group contained Agelaius phoeniceus, Sturnella magna, and S. neglecta. The Quiscaline group contained Euphagus cyanocephalus, Cassidix mexicanus, and Quiscalus quiscula. Molothrus ater, the most divergent, was placed in a separate group. Divergence times for the seven species were compared to the literature. Heterozygosity of the seven populations of the two species of Sturnella were compared to determine factors influencing their divergence. Two factors proposed were heterosis in S. neglecta and possible hybridization between S. neglect and S. magna.
Biology and Energetics of Tropisternus Lateralis Nimbatus (SAY) (Coleoptera: Hydrophilidae) in a Playa on the Southern High Plains of Texas and Aquatic Coleoptera Diversity from Seven Playas on the Southern High Plains of Texas
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.
Biosystematic Study of a Desmodium Complex
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.
Bottom-Up Impacts of Grazing Disturbance on Ground-Nesting Bee Assemblages: Do They Dig It?
In my thesis, I examined impacts of nesting and floral resources on ground-nesting bee assemblages at nine ranch sites with differing grazing histories. Because ecological disturbances can strongly affect the availability of foraging and nesting resources over time, I also examined the impacts of grazing history on nesting and floral resources for ground-nesting bees. I sampled bee assemblages, a comprehensive array of vegetation and soil measures, and floral abundance and richness. I used these data to determine (1) the importance of different nesting habitat resources and livestock grazing history for ground-nesting bees and (2) how livestock grazing history influenced the availability of nesting and floral resources. I found that sites with sandier soils had greater abundance and richness of ground-nesting bees, and sites with less compacted soils had greater bee abundance. Contrary to many other studies, the availability of bare ground was not important for ground-nesting bee abundance or richness. Grazing history did not affect any measures of nesting or floral habitat, and had negligible effects on ground-nesting bee abundance, richness, and community composition. My results suggest that the availability of sandier or less compacted soils may be limiting for ground-nesting bee abundance or richness, especially in areas with predominately clay soil types. In this context, the availability of such nesting resources may be more limiting than floral resource availability. More research is necessary to tease apart the effects of different nesting and floral resources for bees, and how different grassland management types affect bees through those resources. My findings highlight the importance of considering nesting habitat resources in bee conservation and restoration efforts.
Capillary Permeability to Narrow-Range Macromolecular Dextrans at Normal and Hypobaric Pressures
In view of its varied concepts and interpretations, and because of the discrepancies produced by the previous utilization of polydispersed dextrans, a study using extremely narrow-range molecular weight dextran fractions was initiated to reevaluate and consolidate some of the aspects of capillary permeability. A portion of the study was performed under decreased barometric pressure in order to clarify further some of the mechanisms involved in particulate transfer across the capillary endothelial membranes. Gel filtration procedures augmented the study as an assessment of the polydispersity effects of the dextrans employed.
Characterization of Arthrobacter Globiformis Aspartate Transcarbamylase Concentrations of Substrates
This thesis consists of one major section with two subsections. The first subsection investigates the activity of Arthrobacter globiformis aspartate transcarbamylase's specific activity with increasing concentrations of the enzyme's substrate. Dihydroorotase (DHOase) activity was also measured with increasing concentrations of the substrate dihydroorotate. The second subsection collected data in order to classify the enzyme, resulting in a classification into the category of class A ATCases with bifunctional ATCase-DHOase complexes. The thesis provides evidence to broaden understanding of the ATCase and DHOase enzymes for members of the family that Arthrobacter belongs to.
Chemotactic Response of Lumbricus terrestris Coelomocytes to Larval and Adult Stages of Rhabditis pellio
Experiments were performed to assess the suitability of Rhabditis pellio, a nematode found in earthworms, as a challenge organism for use in development of a biomarker assay to determine the potential of chemicals to suppress the immunocompetence of the non-specific immune system. To accomplish this goal, information on the life cycle of R. pellio was determined; including effects of incubation time and temperature on growth rates; along with information on the immune response elicited in the earthworm, Lumbricus terrestris. Immune parameters measured were coelomocyte migration toward and attachment to R. pellio larvae and adults. Preliminary background information showed that R. pellio has potential as a challenge organism for development of a biomarker assay.
Chronic Hypoxia and Hyperoxia Modifies Morphology and Vegf Expression of the Lungs of the Developing Chicken (Gallus Gallus Domesticus)
This study determines effects of oxygen levels on morphology and VEGF expression of developing chicken lungs following incubation in normoxia (21% O2), hypoxia (15% O2) or hyperoxia (30% O2), until developmental days 16 or 18. Lung morphology was assessed using light microscopy, while VEGF expression was determined with ELISA. In hypoxia, the proportion of parabronchial tissue and parabronchi including lumina increased from day 16 to 18 (61 to 68% and 74.2 to 82.2%, respectively). Non-parabronchial tissue was higher in hypoxia than in hyperoxia on day 16 (26 to 20%). However, by day 18, there were no differences between groups. VEGF expression was 33% higher in hypoxia than in hyperoxia on day 16 (736 vs. 492 pg/ml). On day 18, VEGF expression was 43% higher in hyperoxia than in normoxia (673 to 381pg/ml), and remained elevated by 40% in hypoxia over normoxia (631 pg/ml). VEGF may be a mechanism by which parabronchial tissue is stimulated from day 16 to 18 following exposure to chronic hypoxia.
Cigarette Residues Affect Steroidogenesis in Cultured Y-1 Mouse Adrenal Tumor Cells
This study (1) quantitatively compared steroid production in cultured Y-l mouse adrenal tumor cells exposed to Camel and Carlton-smoke derived residues, and (2) localized the effects in the cell. Basal steroid production was increased by Camel residues but not by Carlton, while ACTH stimulation was interfered with by both residues. Camel basal stimulation was comparable to that of cAMP, and was abolished by Cytochalasin D. The stimulation was also comparable to that of cholera toxin, which activates adenyl cyclase. Results indicate that residue components dissolve in the membrane stimulating adenyl cyclase at a point similar to or before that utilized by cholera toxin for its stimulating effect.
Classification of Toolmark Surfaces on Zipper Teeth
This study proposes the classification of the toolmark under the heads of zipper teeth as a subclass characteristic as outlined by the Association of Firearm and Toolmark Examiners (AFTE). Two separate cases in which zipper teeth were found at crime scenes prompted this study. Brass zipper teeth manufactured by YKK were taken from 20 pairs of jeans and studied using a Reichert comparison microscope at 4X power. Photographs were taken and over 750 comparisons made. It was found that the toolmarks on each side on the 20 zippers were unique and independent of all other sides. The observations made in this study indicate that classifying zipper teeth toolmarks as a subclass characteristic is valid.
Coelomic Fluid Protein Profile in Earthworms Following Bacterial Challenge.
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.
Comparative Chemistry of Thermally Stressed North Lake and Its Water Source, Elm Fork Trinity River
To better understand abiotic dynamics in Southern reservoirs receiving heated effluents, water was analyzed before and after impoundment in 330 ha North Lake. Macronutrients, metals, and chlorinated hydrocarbons were measured. Concentrations of nutrients and metals in sediments were quantified in this 2 yr study. River water prior to impoundment contained 16 times more total phosphorus, and supported 23 times more Selenastrum capricornutum cells in an algal assay than reservoir water. The reservoir has essentially no drainage and since evaporation is high, the concentrations of many dissolved solids have increased since the reservoir was filled in 1958. North Lake is now phosphorus limited. Apparently altered chemical equilibria have caused precipitation or adsorption of phosphorus with calcium and iron.
Concentration-dependent Effects of D-Methylphenidate on Frontal Cortex and Spinal Cord Networks in vitro
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.
Data Mining Using Direct Injection Triple Quadrupole Mass Spectrometry, Infrared Spectroscopy, Inductively Coupled Plasma Optical Emission Spectroscopy, and Polymerase Chain Reaction for the Rapid Identification of Nutraceuticals and Contaminants
There has been a rapid surge toward "organic" products devoid of GMOs, MSGs, and other common compounds found in processed foods that continue to indicate an association with an increased risk for disease. These consumers seek nutrients and vitamins that are lacking in their diet and lifestyle in the form of nutraceuticals for disease prevention and treatment as well as overall lifestyle enhancement. However, these products generally lack clinical evidence as well as legal definition. Due to this ambiguity, nutraceuticals are neither considered a food product nor a pharmaceutical product. Furthermore, due to their alleged natural properties allowing for safe, therapeutic effects, nutraceuticals are being eagerly sought after by consumers in the place of pharmaceuticals. Additionally, since nutraceutical substances are "naturally" derived, there is a general lack of regulation regarding the manufacturing and distribution process. This mismanagement leads to lack of quality assurance (QA) and quality control (QC) protocols strictly implemented to define appropriate production and storage parameters. Without these critical measures, consumers are subjected to contamination of their products resulting from improper storage conditions and unmanaged production. These contaminants often include heavy metal impurities, pesticides, bacterial activity, and may also be adulterated with illicit drugs, all leading to detrimental health and environmental effects.
Density, Distribution, Production and Drift of Benthic Fauna in a Reservoir Receiving Thermal Discharges from a Steam Electric Generating Plant
The effects of thermal discharges on the density, distribution, production, and drift of benthic organisms were studied at North Lake reservoir.
Determination of Biotransformation and Biodegradation Rate Constants for Naphthalene, Lindane and Phenol
Biotransformation and biodegradation rate constants were determined for naphthalene, lindane, and phenol in water samples from three different sources. Rate constants produced from monitoring disappearance of the parent chemical (biotransformation) were compared to those obtained from mineralization of the chemical (ultimate biodegradation) by ¹⁴CO₂ evolution as well as acidification of the residual ¹⁴C-labeled compound (primary biodegradation). Rate constants were statistically different for the three chemicals. The water source affected the rate constants. When biomass measurements of the waters were considered and second-order rate constants were derived, there was no statistical evidence that this parameter gave a reliable rate constant statistic that could be useful in predicting the fate of any of naphthalene, lindane, and phenol in these waters.
Determination of Dissociation Constants for GABAA Receptor Antagonists using Spontaneously Active Neuronal Networks in vitro
Changes in spontaneous spike activities recorded from murine frontal cortex networks grown on substrate-integrated microelectrodes were used to determine the dissociation constant (KB) of three GABAA antagonists. Neuronal networks were treated with fixed concentrations of GABAA antagonists and titrated with muscimol, a GABAA receptor agonist. Muscimol decreased spike activity in a concentration dependent manner with full efficacy (100% spike inhibition) and a 50% inhibitory concentration (IC50) of 0.14 ± 0.05 µM (mean ± SD, n=6). At 10, 20, 40 and 80 µM bicuculline, the muscimol IC50 values were shifted to 4.3 ± 1.8 µM (n=6), 6.8 ± 1.7 µM (n=6), 19.3 ± 3.54 µM (n=10) and 43.5 µM (n=2), respectively (mean ± SD). Muscimol titration in the presence of 10, 20, 40 µM of gabazine resulted in IC50s values of 20.1 (n=2), 37.17 (n=4), and 120.45 (n=2), respectively. In the presence of 20, 80, and 160 µM of TMPP (trimethylolpropane phosphate) the IC50s were 0.86 (n=2), 3.07 (n=3), 6.67 (n=2) µM, respectively. Increasing concentrations of GABAA antagonists shifted agonist log concentration-response curves to the right with identical efficacies, indicating direct competition for the GABAA receptor. A Schild plot analysis with linear regression resulted in slopes of 1.18 ± 0.18, 1.29 ± 0.23 and 1.05 ± 0.03 for bicuculline, gabazine and TMPP, respectively. The potency of antagonists was determined in terms of pA2 values. The pA2 values were 6.63 (gabazine), 6.21 (bicuculline), and 5.4 (TMPP). This suggests that gabazine has a higher binding affinity to the GABAA receptor than bicuculline and TMPP. Hence, using spike rate data obtained from population responses of spontaneously active neuronal networks, it is possible to determine key pharmacological properties of drug-receptor interactions.
Development of a Watershed Prioritization Strategy Using Remote Sensing and the Geographic Resource Analysis Support System. Case study: City of Carrollton, Texas
This study used a geographic information system (GIS) and an empirical pollutant load model to estimate average annual pollutant loads contained in storm water charges from the City of Carrollton, Texas. The Geographic Resource Analyses Support System (GRASS) was used to create a number of spatial data layers as input variables to a model that estimates loads in Kilograms/Hectare/Year, and Kilogram/Basin/Year. Average annual pollutant loads for twelve pollutants were derived from average annual rainfall/runoff volumes, coupled with Local Dallas/Fort Worth Metroplex storm water Event Mean Concentrations.
Diphenyloxazole Metabolism by Aryl Hydrocarbon Hydroxylase
2,5-Diphenyloxazole (PPO) was tested as a potential alternate inducer for the aryl hydrocarbon hydroxylase (AHH) system. Its apparent lack. of carcinogenicity and toxicity provide a possible system for investigation of enzyme systems related to chemical carcinogenesis without exposure of the researcher to potent carcinogenic compounds. These studies found PPO to be an inducer of AHH in cultured human lymphocytes. When PPO was utilized as a substrate for the AHH assay system, the major metabolites produced were strongly fluorescent. A simple fluorometric assay was developed which employed PPO as the substrate and which measured constitutive activity more efficiently than similar assays using benzo(a)pyrene as the substrate. Quantitation of both basal and induced lymphocyte AHH metabolism of PPO may be applicable to human population studies and may provide a tool to determine possible genetic variables with respect to carcinogen metabolism related to cancer risk.
Distribution, Abundance, and Food Habits of Larval Fish in a Cooling Reservoir
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.
Distribution of a Novel Gram Negative, Capsule-Forming Bacterium
A novel Gram negative, capsule-forming bacterium was previously isolated in Dr. G. Roland Vela's laboratory. The distribution of this bacterium in soils from various locations was investigated. Soil samples from 188 locations around the world were examined. Isolates of the bacterium were obtained from 50 of these soils, with 48 of the isolates found in soils from the southwestern United States and northern Mexico. This suggests that this region is the natural habitat of the bacterium. The other two isolates were obtained from Madrid, Spain and Taipei, Taiwan. None were found in soils from South America or Australia. A lack of variation in morphology and physiological properties in the isolates suggests that a homogeneous population exists, even from widespread geographical locations.
Distribution, Size, Condition, and Food Habits of Selected Fishes in a Reservoir Receiving Heated Effluent from a Power Plant
This study was undertaken in order to provide further insight into the effects of artificial heating on the fisheries of a small reservoir in the Southwest. The following specific objectives were established: (1) to map the reservoir for the distribution of heated water, (2) to determine the distribution of selected species of sports, rough, and forage fish in areas affected by the effluent and in areas not affected, (3) to compare size and condition of selected species of fish from areas affected by the effluent to size and condition of fish from areas not affected, and (4) to compare food habits of channel catfish in areas affected by the effluent to the food habits of channel catfish in areas not affected.
DNA-DNA Hybridization of Methane Oxidizing Bacteria
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.
Drumming Behavior of Selected North American Stoneflies (Plecoptera)
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.
The Dynamics of a Planktonic Microcrustacean Community in a Small North Central Texas Pond Ecosystem
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.
Ecological Association Between the Red-Cockaded Woodpecker and Southern Pine Beetle in the Homochitto National Forest: a Geographic Information System Approach
Since the introduction of management practices by the Forest Service to stabilize red-cockaded woodpecker (RCW) populations, the number of cavity trees killed by southern pine beetles (SPB) has increased. A model of the landscape ecology of RCW and SPB in the Homochitto National Forest was created using data collected from the Forest Service and Global Atmospherics. The conclusions of the study were that the RCW and SPB utilize the same type of habitat and the stand hazard maps are an accurate means of determining the locations of SPB infestations. The functional heterogeneity maps created for the SPB and RCW would be useful predictors of future occurrences of either species if complete data were obtained.
The Effect of Menthol on Nicotine Metabolism: a Cross Species Evaluation
The effect of menthol on nicotine metabolism was examined in liver S9 fractions of four different species and in the in vivo mouse model. The purpose of this study was to investigate three parameters: (1) biotransformation of nicotine to cotinine in various species (human, mouse, rat and trout) using in vitro methods; (2) to determine if the addition of menthol with nicotine altered biotransformation of nicotine to cotinine; (3) and to assess similar parameters in an in vivo mouse model. The major findings of this study include: (1) mice appear to metabolize nicotine, over time, in a manner similar to humans; (2) menthol decreased cotinine production, over time, after a single dose in mice; and (3) menthol increased cotinine production, over time, after repeated doses, in mice.
The Effect of Post-exercise Ethanol Consumption on the Acute Hormonal Response to Heavy Resistance Exercise in Women
The purpose of this study was to examine the hormonal response to acute ethanol ingestion following a bout of heavy resistance exercise in women. Eight resistance trained women completed two identical acute heavy resistance exercise tasks (AHRET). From 10-20 minutes post-AHRET, participants consumed either a grain ethanol or a placebo beverage. Blood was collected before (PRE) and immediately after the AHRET (IP) and then every 20 minutes for five hours. Blood collected after beverage ingestion was pooled into 3 batches (phases: 20-40 minutes, 60-120 minutes, and 140-300 minutes post-exercise) and analyzed for serum total testosterone (TT), free testosterone (FT), insulin-like growth factor-I (IGF-I), human growth hormone (GH), cortisol (COR), and estradiol (E2) concentrations. Circulating concentrations of TT were significantly greater at P20-40 than at PRE, P60-120, and P140-300. Circulating concentrations of FT were significantly greater at P20-40 than at all other times. Circulating concentrations of GH were significantly greater at IP than at PRE, P60-120, and P140-300. Circulating concentrations of COR were significantly greater at P20-40 than at all other times. Additionally, COR concentrations at P140-300 were significantly lower than at all other times. Circulating concentrations of IGF-1 were significantly greater at P20-40 than at P60-120 and P140-300. Circulating concentrations of E2 were significantly greater at P20-40 than at all other times. In summary, the present study demonstrated an acute modulation of the neuroendocrine milieu following a heavy resistance exercise bout in women. Ethanol ingestion appeared to have no significant effect on the characteristics of acute hormonal augmentation in TT, FT, GH, COR, IGF-1, or E2.
The Effects of 3-Deazaguanine on Chick Embryo Fibroblasts and Rat Kidney Cells Infected with Temperature-Sensitive Mutant and Wild-Type Rous Sarcoma Viruses
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.
Effects of Airway Pressure, Hypercapnia, and Hypoxia on Pulmonary Vagal Afferents in the Alligator (Alligator Misssissippiensis)
The American alligator (Alligator mississippiensis) is an aquatic diving reptile with a periodic breathing pattern. Previous work has identified pulmonary stretch receptors (PSR), both rapidly- and slowly-adapting, and intrapulmonary chemoreceptors (IPCs) that modulate breathing patterns in alligators. The purpose of the present study was to identify the effects of prolonged lung inflation and deflation (simulated dives) on PSR and/or IPC firing characteristics in the alligator. The effects of airway pressure, hypercapnia, and hypoxia on dynamic and static responses of pulmonary stretch receptors (PSR) were studied in juvenile alligators (mean mass = 246 g) at 24°C. Receptor activity appeared to be a mixture of slowly-adapting PSRs (SARs) and rapidly-adapting PSRs (RARs) with varying thresholds and degrees of adaptation, but no CO2 sensitivity. Dives were simulated in order to character receptor activity before, during, and after prolonged periods of lung inflation and deflation. Some stretch receptors showed a change in dynamic response, exhibiting inhibition for several breaths after 1 min of lung inflation, but were unaffected by prolonged deflation. For SAR, the post-dive inhibition was inhibited by CO2 and hypoxia alone. These airway stretch receptors may be involved in recovery of breathing patterns and lung volume during pre- and post-diving behavior and apneic periods in diving reptiles. These results suggest that inhibition of PSR firing following prolonged inflation may promote post-dive ventilation in alligators.
Effects of External Electric Fields on Light Transmittance in Isolated Crayfish Nerves
Acute effects of a pulsed external electric field (PEEF) at 20 V/cm and a d.c. EEF at 90 V/cm on light transmittance in an isolated compound crayfish nerve was measured. In a third series, the nerve was pre-treated with the Na+ channel blocker tetrodotoxin (TTX). A PEEF produced an irreversible increase in the variation of light transmittance in normal nerves but a reversible increase in TTX treated nerves. This data was statistically insignificant. The d.c. EEFs produced a reversible and statistically significant enhancement of variation in light transmittance in both untreated and TTX-treated nerves. The findings may be due to either (1) an alteration in the ion/fluid flux within the nerve or (2) a physical alteration of protein molecules in the membranes.
Effects of High Altitude Exposure on Capillary Permeability
Observations of decreases in plasma volume, shifts in plasma and lymph protein concentrations, and increases in capillary permeability at high altitude have been reported in the literature by several investigators. This investigation was begun in an attempt to elucidate the possible significance of these phenomena in future space exploration, and because of the lack of knowledge concerning the underlying mechanisms. The purpose of this investigation was to determine the effects of exposure to hypobaric pressures on the capillary permeability to the normal plasma and lymph proteins.
Effects of Long-Term Moderate Ethanol Intake on the Stress Response in Rats
The effect of ethanol on the stress response in rats was examined. Experimental animals were given 0.25 ml of 28 percent ethanol or 0.25 ml of water orally once a day, five days a week, for a period of twelve months and were then subjected to fifteen minute cold stress. Corticosterone levels in ethanol-treated males following stress were significantly lower (22 percent) than in the sham group. Adrenal weights in sham-treated females were significantly higher (15 percent) than in the ethanol group at the end of twelve months. Mortality in sham-treated males was significantly higher (60 percent) than in ethanol-treated males. The effects observed may be due to the sedative action of ethanol on cortical centers controlling the hypothalmus.
The Effects of Long Term Modernate Ethanol Intake on Plasma Levels of ACTH, Beta Endorphin, and Corticosterone in Rats
The effects of single injections and daily oral administration of ethanol on plasma levels of ACTH, beta endorphin, and corticosterone in response to cold stress were examined. The long-term experimental animals were given 0.25 ml of 28% ethanol or water orally once a day, five days a week, for fourteen months. Plasma levels of ACTH, beta endorphin, and corticosterone were lower in alcohol-treated rats as compared with water-treated rats when exposed to cold stress. The effects of a single injection of ethanol significantly elevated plasma levels of all three hormones. Mortality in sham-treated males was higher than ethanol-treated.
Effects of N⁶,O²'-Dibutyryl Cyclic Adenosine 3' ,5' Monophosphate on Transformation of Rat Kidney Cells and Chick Embryo Fibroblasts by Wild-Type and Temperature-Sensitive Rous Sarcoma Virus
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).
Effects of Sertraline Exposure on Fathead Minnow (Pimephales promelas) Steroidogenesis
Sertraline is a selective serotonin reuptake inhibitor (SSRI) that is widely used for the treatment of depression and anxiety. Due to the abundant therapeutic use of sertraline, low levels have been detected in municipal wastewater effluents suggesting that aquatic organisms may be exposed. The purpose of this study was to evaluate the steroidogenic effects of sertraline on larval (FHM) and adult female fathead minnows (FFHM), Pimephales promelas. Larval FHM were exposed to 0.1, 1, and 10 µg/L sertraline for 28 days and analyzed via RT-qPCR for differential expression of 11β-Hydroxysteroid dehydrogenase (11β-HSD), 20β-Hydroxysteroid dehydrogenase (20β-HSD), aromatase (CYP19), and nuclear thyroid receptor alpha (TRα). FFHM were exposed to 3 or 10 µg/L sertraline for 7 days with the brain and ovary excised at exposure termination. Juvenile FHM exposed to 0.1 μg/L sertraline had a significant upregulation of both 20β-HSD and TRα. FFHM exposed to 10 µg/L sertraline had a significant upregulation of 11β-HSD expression in brain tissue, while no steroidogenic changes were observed in the FFHM ovary. Similarly, in FFHM brain tissue, CYP19 and 20β-HSD expression levels were significantly higher in fish exposed to 10µg/L sertraline compared to control. The significance of these findings with respect to survival, growth and reproduction are currently unknown, but represent future research needs.
Endocannabinoid System in a Planarian Model
In this study, the presence and possible function of endocannabinoid ligands in the planarian is investigated. The endocannabinoids ananadamide (AEA) and 2-arachidonoylglycerol (2-AG) and entourage NAE compounds palmitoylethanolamide (PEA), stearoylethanolamide (SEA) and oleoylethanolamide (OEA) were found in Dugesia dorotocephala. Changes in SEA, PEA, and AEA levels were observed over the initial twelve hours of active regeneration. Exogenously applied AEA, 2-AG and their catabolic inhibition effected biphasic changes in locomotor velocity, analogous to those observed in murines. The genome of a close relative, Schmidtea mediterranea, courtesy of the University of Utah S. med genome database, was explored for cannabinoid receptors, none were found. A putative fatty acid amide hydrolase (FAAH) homolog was found in Schmidtea mediterranea.
Evaluation of Diet, Water, and Culture Size for Ceriodaphnia Dubia Laboratory Culturing
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.
The Evolutionary Genetics of Campostoma anomalum and Campostoma oligolepis
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 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 their genetic integrity throughout their ranges. their genetic integrity throughout their ranges.
Evolutionary Genetics of Three Semispecies of Wood Rats--Neotoma Albigula, Neotoma Micropus, and Neotoma Floridana
Electrophoretic variation in 18 proteins encoded by 20 autosomal loci was used to compare the genetic relationships of 19 natural populations representing three species of the subgenus Neotoma. Of the 20 loci examined nine were monomorphic and fixed for the same allele in all populations. No more than seven loci were polymorphic within a single population. Genetic variability was expressed as the proportion of loci heterozygous in the average individual of a population. Heterozygosity in the three species of Neotoma studied averaged 0.078, a value within the range reported for other rodents. Although the levels of heterozygosity seen in Neotoma could not readily be explained, the variation may be attributed to ecological factors. The three species of Neotoma were compared on the basis of genetic similarity and found to form a close taxonomic unit, probably semispecies. Divergence times were obtained for the three species and found to compare well with divergence times obtained from fossil data. In general, the three species have diverged within the last 112,000 years during the Wisconsin glacial period.
Experimental Trichinosis in Birds
This work concerns itself with essentially four experiments: (1) the cecum-injective-infection experiment; (2) the anus-injective-infection experiment; (3) the mouth ingestive-infection with larvae, and (4) the mouth ingestive-infection with the flesh of infected rats.
Gene Dosage Study on Human Chromosome 22
A gene dosage study was conducted on a rare complete trisomy 22 human fibroblast cell line utilizing three lysosomal enzymes, ∝-iduronidase, ∝-galactosidase B, and arylsulfatase A, whose genes are located on chromosome 22 and two control enzymes, ,β-hexosaminidase A and -- fucosidase, with genes not on chromosome 22. A gene dosage effect was clearly demonstrated for an early passage number of the fibroblasts; however, later passage numbers gave inconclusive results. This study suggests that gene dosage studies must be carefully designed to be conducted only on early, matched passage number cells. ∝-fucosidase gave anomalous results most likely due to pleiotropic effects. The present gene dosage study confirmed the trisomic nature of the cell line studied and suggests that this type of study may be a useful diagnostic tool for small deletions, additions, or unbalanced translocations.
Genic Differentiation and Evolution in the Ground Squirrel Subgenus Ictidomys (Spermophilus)
The genetic structure of 26 natural populations of three species (S. tridecemlineatus, S. mexicanus, and S. spilosoma) of the Ictidomys subgenus of ground squirrels was analyzed using chromosomal and electrophoretic techniques. Chromosomal variation was not observed in S. mexicanus, and only slight karyotypic variation was found in the other two species. Chromosomal evidence indicated hybridization between S. tridecemlineatus and S. mexicanus, placing these species within the classical definition of semispecies. Analysis of electrophoretic variation at 29 genetic loci indicated close genetic relationships between these species. Evolution in Ictidomys appears to be linked with Pleistocene events, and speciation appears to have occurred within the last 155,000 years.
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