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Analysis of Phytoplankton Responses to Water Chemistry Dynamics in a Moderately Eutrophic North Texas Reservoir
Canonical correspondence analysis (CCA) was used to explore relationships between measured environmental variables and in situ phytoplankton communities in a moderately eutrophic North Texas Reservoir.
L-asparaginase II Production by Escherichia coli
Growth of Escherichia coli A-l under aerobic conditions in an enriched medium with a total amount of 0.2 per cent glucose was biphasic and asparaginase II activity was detected after depletion of ammonia from the growth medium in the second phase of growth. Glucose was exhausted two hours before ammonia and three hours before asparaginase II activity was detected. The concentration of 3',5'-cyclic adenosine monophosphate was found to fluctuate when the dissolved oxygen in the medium reached a low level, when glucose and ammonia were exhausted, and when the cells entered the second stationary phase of growth. Culture tube studies of the growth of E_j_ coli A-l in three per cent nutrient broth with varied concentrations of ammonium chloride and potassium nitrate gave lower specific activity of asparaginase II when this was compared to that seen in three per cent nutrient broth alone. The addition of glucose to the same medium before asparaginase II activity was detected resulted in the production of acid by E. coli A-l with cessation of growth; however, addition after L-asparaginase synthesis had started did not affect the specific activity of the enzyme. The addition of ammonium chloride suppressed L-asparaginase synthesis, but addition after enzyme synthesis started had no affect. These findings suggest that asparaginase II is produced by E. coli A-l in response to low concentrations of ammonia and that exogenously supplied nitrogen compounds may play a major role in the regulation of this enzyme. It is suggested that E. coli A-l produced L-asparaginase in order to obtain ammonia for the synthesis of glutamine from glutamate. The synthesis of glutamine from glutamate is the first step of a highly branched pathway which ultimately leads to the synthesis of many of the important macromolecules of the cell.
Biological and Toxicological Responses Resulting from Dechlorination of a Major Municipal Wastewater Treatment Plant Discharge to the Trinity River
Federal regulations such as the Clean Water Act (P.L. 92-500), and its amendments, direct the Environment Protection Agency (EPA) to implement programs to control the releases of conventional pollutants and toxics into the waterways of the United States. The EPA began requiring treatment plants to conduct toxicity tests (biomonitoring) of their effluent discharges. To control toxicity caused by chlorination of wastewater discharges, the EPA also began requiring some treatment facilities to dechlorinate their wastewater before discharging. This research was funded by the EPA to document the changes that occurred in the Trinity River from the dechlorination of the effluent from Ft. Worth's Village Creek municipal wastewater treatment plant. The study occurred over a two year period beginning in August 1990. A wide variety of biological field assessments and toxicological assays were used to measure various responses. Seven river stations, covering approximately twenty river miles, and the treatment plant effluent were assessed. Two of the river stations were upstream from the treatment plant and used as reference sites. The remaining five river stations were downstream from the treatment plant, spread out over seventeen river miles. The study evaluated the impact of chlorination prior to dechlorination, which served as a baseline. Responses determined during dechlorination were compared to the baseline data. An overall improvement in species richness and diversity was seen at those river stations which had previously been adversely impacted by chlorine. Aquatic toxicity tests, such as those required to be used by dischargers, were conducted during this study. Periodic toxicity was observed with these tests in the effluent and river samples after dechlorination was initiated. Those tests, along with in situ toxicity assays, proved to be good predictors of biological community responses.
Cardiovascular Fetal Programming in Quail (Colinus virginianus), An Avian Comparative Model
The consequences of early embryonic insults and how they affect subsequent life reflects the emerging concept of "fetal programming". The aim of this project is to study the effects of embryonic insults as they subsequently manifest themselves in adults, with emphasis on the heart and vasculature. My experiments establish that fetal programming operates on the bobwhite quail inducing similar changes as those observed in mammalians and other birds. The quail's fast development provides reliable data in a short period of time than other avian models (e.g. domestic chicken). Data on quail showed a correlation between egg mass and hatchling mass; where small eggs produce small hatchlings but a high mortality made it impractical as a stressor for this study. Hypoxia was used as a stressor during embryonic incubation, where it induced a low hatching weight in quail that was not observable in adult birds. Morphological measurements demonstrated an increased ventricular collagen content and reduced ventricular lumen in birds in adults incubated in hypoxia consistent with hypertension. The hematological analyzes showed few differences indicating organ remodeling instead of hematopoietic compensation. The assessment of vascular reactivity pointed out an impaired endothelium dependent relaxation commonly associated to hypertension in birds and mammals. Fetal programming could be a widespread response to an adverse prenatal environment in endotherms and the resulting data from this work contributes to our understanding of fetal programming in vertebrates and its long term consequences.
Construction of a Pseudomonas aeruginosa Dihydroorotase Mutant and the Discovery of a Novel Link between Pyrimidine Biosynthetic Intermediates and the Ability to Produce Virulence Factors
The ability to synthesize pyrimidine nucleotides is essential for most organisms. Pyrimidines are required for RNA and DNA synthesis, as well as cell wall synthesis and the metabolism of certain carbohydrates. Recent findings, however, indicate that the pyrimidine biosynthetic pathway and its intermediates maybe more important for bacterial metabolism than originally thought. Maksimova et al., 1994, reported that a P. putida M, pyrimidine auxotroph in the third step of the pathway, dihydroorotase (DHOase), failed to produce the siderophore pyoverdin. We created a PAO1 DHOase pyrimidine auxotroph to determine if this was also true for P. aeruginosa. Creation of this mutant was a two-step process, as P. aeruginosa has two pyrC genes (pyrC and pyrC2), both of which encode active DHOase enzymes. The pyrC gene was inactivated by gene replacement with a truncated form of the gene. Next, the pyrC2 gene was insertionally inactivated with the aacC1 gentamicin resistance gene, isolated from pCGMW. The resulting pyrimidine auxotroph produced significantly less pyoverdin than did the wild type. In addition, the mutant produced 40% less of the phenazine antibiotic, pyocyanin, than did the wild type. As both of these compounds have been reported to be vital to the virulence response of P. aeruginosa, we decided to test the ability of the DHOase mutant strain to produce other virulence factors as well. Here we report that a block in the conversion of carbamoyl aspartate (CAA) to dihydroorotate significantly impairs the ability of P. aeruginosa to affect virulence. We believe that the accumulation of CAA in the cell is the root cause of this observed defect. This research demonstrates a potential role for pyrimidine intermediates in the virulence response of P. aeruginosa and may lead to novel targets for chemotherapy against P. aeruginosa infections.
Ecotoxicological Investigations in Effluent-Dominated Stream Mesocosms
The University of North Texas Stream Research Facility (UNTSRF) was designed to examine contaminant impacts on effluent-dominated stream ecosystems. Stream mesocosms, fed municipal effluent from the City of Denton, TX, Pecan Creek Water Reclamation Plant (PCWRP), were treated with 0, 15 or 140 µg/L cadmium for a 10-day study in August 2000. Laboratory toxicity test and stream macroinvertebrate responses indicated that cadmium bioavailability was reduced by constituents of effluent-dominated streams. The Biotic Ligand Model (BLM) for Cd was used to predict a 48 hour Cd EC50 for Ceriodaphnia dubia of 280 µg/L in these effluent-dominated streams. This value is higher that an EC50 of 38.3 µg/L Cd and a 7-day reproduction effect level of 3.3 µg/L Cd generated for C. dubia in reconstituted laboratory hard water. These results support use of a cadmium BLM for establishing site-specific acute water quality criteria in effluent-dominated streams. Although not affected by 15 µg/L treatments, organisms accumulated Cd in 15 µg/L treated streams. Hence, over longer exposure periods, Cd accumulation may increase and a no effect level may be lower than the observed 10-day no effect level of 15 µg/L. A toxicity identification evaluation procedure was utilized with in vitro and in vivo bioassays to identify estrogenic compounds in PCWRP effluent, previously identified to seasonally induce vitellogenin (VTG) in male fathead minnows. Steroids, nonylphenol ethoxylate metabolites, and other unidentified compounds were identified as causative effluent estrogens. These findings suggest that in vivo VTG bioassays should be used to confirm in vitro Yeast Estrogen Screening assay activity when effluents are fractionated or screened for estrogenicity. A subsequent 90-day cadmium study was initiated to assess long-term effluent and cadmium effects on fish endocrine function. Juvenile fathead minnows were placed in UNTSRF pool sections of replicate streams treated with 0, 5, 20 or 80 µg/L Cd. Male …
Effects of 4-Chloroglutaranilic Acid on Growth and Development of Sunflower Seedlings
The potential growth-regulating compound 4-chloroglutaranilic acid (CGA) was tested in whole-plant bioassay systems which utilized sunflower seedlings (Helianthus annuus, L.). Test systems included the growth of plants in soil , solid inert (Vermiculite) substrate, and hydroponic (Seed-Pak) pouches.
The Effects of Organic Surface Amendments on Soil Nutrients and Initial Tree Establishment
This study examined the effects of replicating woodland soil surface horizonation on the nutrient status of underlying soils and the initial establishment and growth of trees. A total of 283 container grown trees were planted in a bufferzone around a future landfill site. Control amendments consisted of an 8 cm layer (0.5 m3) of wood chips applied in a circular area of 4.6 m2 around the trees' planting pit. For the treatment, a 2.5 cm layer of composted biosolids (0.15 m3 or 80 Mg/ha) was applied in a circular area of 4.6 m2 around the trees' planting pit followed by an 8 cm layer (0.5 m3) of wood chips. The results indicate that the replication of woodland soil surface attributes using composted biosolids can significantly improve the nutrient status of underlying soil. Some significant effects were seen under control conditions, too. However, the effects on tree establishment and growth parameters were, for the most part, not statistically significant.
Eutrophic Levels of Different Areas of a Reservoir: A Comparative Study
It was the purpose of this investigation to attempt to demonstrate if differences in eutrophic levels existed among selected areas of Garza-Little Elm, and to demonstrate the role that sediments play in affecting eutrophication.
A Floristic Study of the Woody Vegetation of the North American Cross Timbers
This research represents the first systematic collection of the woody plants throughout the Cross Timbers. It provides the first keys to these plants in their vegetative condition, plant descriptions, distribution maps, and some quantitative measurements used for descriptive purposes. Descriptions of the woody plants were constructed as an aid in verification after a specimen has been identified by use of the keys. The measurements given pertain only to the woody plants as they occur in the Cross Timbers. Distributional maps are provided for all the taxa considered in this research. With the exception of those species which have the ecological amplitude to grow throughout the Cross Timbers, the distribution of the majority of the remaining species seems to be most strongly influenced by average annual precipitation. In a few instances, conditions associated with latitude appear to govern the distribution of species or varieties within the Cross Timbers. Throughout the Cross Timbers, post oak (Quercus stelta), blackjack oak (Quercus marilandica), and hickory (Caraa texan) dominate the upland forests. The streamside forests are dominated by willow (alix nigra), cottonwood (Populus deltoides), and hackberry (Celtis laevi ata). The variation in the vegetation of the Cross Timbers is not due to any change in dominant species, but rather to the distribution of the associated species which occur in the two prominent community types.
Forest Landscape Dynamics: a Semi-Markov Modeling Approach
A transition model (MOSAIC) is used to describe forest dynamics at the landscape scale. The model uses a semi-Markov framework by considering transition probabilities and Erlang distributed holding times in each transition. Parameters for the transition model are derived from a gap model (ZELIG). This procedure ensures conceptual consistency of the landscape model with the fine scale ecological detail represented by the forest gap model. Spatial heterogeneity in the transition model is driven by maps of terrain with characteristics contained in a Geographic Information System (GIS) database. The results of the transition model simulations, percent cover forest type maps, are exported to grid-maps in the GIS. These cover type maps can be classified and used to describe forest dynamics using landscape statistics metrics. The linkage model-GIS enhances the transition model spatial analytical capabilities. A parameterization algorithm was developed that takes as input gap model tracer files which contain the percent occupation of each cover type through time. As output, the algorithm produces a file that contains the parameter values needed for MOSAIC for each one of the possible transitions. Parameters for the holding time distribution were found by calculating an empirical estimate of the cumulative probability function and using a non-linear least squares method to fit this estimate to an Erlang distribution. The algorithm provided good initial estimates of the transitions parameters that can be refined with few additional simulations. A method for deriving classification criteria to designate cover types is presented. The method uses cluster analysis to detect the number and type of forest classes and Classification and Regression Tree (CART) analysis to explain the forest classes in term of stand attributes. This method provided a precise and objective approach for forest cover type definition and classification. The H. J. Andrews forest in Oregon was used to demonstrate the …
A geospatial tool for assessing potential wildland fire risk in central Texas.
Wildland fires in the United States are not always confined to wilderness areas. The growth of population centers and housing developments in wilderness areas has blurred the boundaries between rural and urban. This merger of human development and natural landscape is known in the wildland fire community as the wildland urban interface or WUI, and it is within this interface that many wildland fires increasingly occur. As wildland fire intrusions in the WUI increase so too does the need for tools to assess potential impact to valuable assets contained within the interface. This study presents a methodology that combines real-time weather data, a wildland fire behavior model, satellite remote sensing and geospatial data in a geographic information system to assess potential risk to human developments and natural resources within the Austin metropolitan area and surrounding ten counties of central, Texas. The methodology uses readily available digital databases and satellite images within Texas, in combination with an industry standard fire behavior model to assist emergency and natural resource managers assess potential impacts from wildland fire. Results of the study will promote prevention of WUI fire disasters, facilitate watershed and habitat protection, and help direct efforts in post wildland fire mitigation and restoration.
High-Resolution Autoradiography with 33p
The purpose of this study is to provide groundwork for future autoradiographic investigations using 33P as a tracer.
The Immune Response of Guinea Pigs as Influenced by Hypobaric Pressure and Normoxic Environment (Part I); Membrane Filter-Fluorescent-Antibody Method for Detection and Enumeration of Bacteria in Water (Part II)
In this work experimental design and tests were established to determine whether antibody production in guinea pigs injected with a bacterial antigen is Influenced by the environment of simulated high altitude with normoxic conditions. Hematological and electrophoretic studies were simultaneously run with the antibody determinations as a check on related responses of the animals.
Immunofluorescence as a Method for the Rapid Identification of Streptococcus Faecalis in Water
The development and refinement of FA has been adequately investigated with major emphasis on pathogenic microorganisms. The development of this technique has reduced both the time and number of biochemical tests necessary to identify a diversity of organisms. The organisms included are the protozoans, as described by Goldman (1953 and 1957) and by Ingram (1961), viruses, as reported by Liu (1955a) and Burgdorfer and Lackman (1960a), pathogenic bacteria which have been investigated by Moody, Goldman, and Thomason (1956), Moody and Winter (1959), Deason, Falcone, and Harris (1957) and Thomason, Cherry, and Moody (1957). Various fungi have been studied with FA by Kaufman and Brandt (1964), Kaufman and Kaplan (1961 and 1963) and Gordon (1958). Therefore, due to the success of the fluorescent antibody technique in many areas of microbiology in previous investigations, the logical assumption was that immunofluorescence might be incorporated into an efficient system in which a specific organism associated with fecal pollution, such as S. faecalis, could be rapidly identified. Based on this assumption, the feasibility of fluorescent antibody techniques, using S. faecalis was investigated as a means of rapid determination of bacterial pollution in water. Although much progress has been achieved in the study of cytochemical reactions by immunofluorescence, no attention has been focused on the application of this method as a determinative tool by which water contamination, due to the presence of the enterococci, could be demonstrated. Specifically, the purpose of the research reported in this dissertation was to devise an applicable, valid, and rapid method that could be employed in the detection and identification of S. faecalis.
Impact of a Genetically Engineered Probiotic Therapy and IGF-1 Genomics in the PAHenu2 Mouse Model of PKU
Absence of functional phenylalanine hydroxylase results in phenylketonuria (PKU). Viable treatments remain few, expensive and secondary conditions such as osteopenia occur in most PKU patients. Objective 1: Given the recently described roles of gut microbes to aid host digestion, an orally administered genetically engineered probiotic as the delivery vehicle for enzyme replacement therapy was created. The engineered probiotic, pHENOMMenal, produced phenylalanine ammonia lyase with significant production of trans-cinnamate (phenylalanine cleavage product) in vitro and resulted in a reduction of 515 μM in blood phenylalanine when fed to PKU animals for 14 days (from 2307µM ± 264µM to 1792µM ± 261µM, n = 6, P < 0.05). The control probiotic produced no change in blood phenylalanine. Thus, pHENOMMenal treatment in PKU mice demonstrated engineered microbes could compensate for a metabolic deficiency of the host. Objective 2: Evaluate the PAHenu2 mouse model of PKU for a genetic discrepancy causing ocular enlargement and delayed development observed only after the PAHenu2 mutation was crossed to the C57BL/6J mouse. When compared to healthy littermates, ELISA indicated a consistent but insignificant decrease in plasma IGF-1 and an increase in ocular IGF-1 in PKU animals. SNP screening demonstrated a differential inheritance of IGF-1 alleles in healthy and PKU animals based on PAH allele inheritance. Ocular and developmental phenotypes in the PAHenu2 colony match those described in previous IGF-1 studies. Understanding the IGF-1 inheritance discrepancy will enable better osteopenia research using PAHenu2 mice and allow breeding of a healthier mouse colony for continued research. Collectively the results from this work describe a new therapeutic approach for treatment of PKU as well as a better understanding of the PAHenu2 mouse model to study this disease.
The Impact of Developmental Stress on Cardiovascular Physiology of Two Archosaur Species: American Alligator (Alligator mississippiensis) and Domestic Chicken (Gallus gallus)
Crocodilians and birds comprise sister taxa of archosaurs, the development of these vertebrates occurs within an egg case that leaves developing embryos susceptible to fluctuations in the nesting environment. Studies suggest that sub-optimal conditions alter morphological growth and cardiovascular physiology. Regulation of the cardiovascular system is immature in the subjects studied, and embryos may rely on humoral rather than neural control of the cardiovascular system. The primary focus of this dissertation was to assess regulatory mechanisms responsible for maintenance of arterial pressure and heart rate. Dehydration stress had marked effects on embryo growth, and altered baseline cardiovascular parameters, while leaving the response to humoral regulator, angiotensin II (Ang II), unaffected. However, dehydrated alligator embryos developed cholinergic tone on heart rate. Hypoxic incubated chicken embryos were reduced in embryo mass, and altered response to humoral regulatory components Ang I and adenosine in addition identifying a novel regulatory component of the cardiovascular response to acute hypoxia. Collectively, these studies add to the existing knowledge of cardiovascular physiology in embryonic archosaurs and suggest that some components of cardiovascular regulation are plastic following developmental stress.
Impacts of the Pyrethroid Insecticide Cyfluthrin on Aquatic Invertibrate Populations in Outdoor Experimental Tanks
The chemical fate and biological impacts of cyfluthrin in aquatic ecosystems were investigated using microcosms (1.9 m^3 concrete tanks) during 1989. Results were compared to a concurrent pesticide registration study using mesocosms (634.7 m^3 earthen ponds). Ten spray drift and five soil runoff simulations were conducted. Pesticide loadings were scaled by system volume, with the same experimental design in ponds and microcosms. Aqueous cyfluthrin concentrations and sediment residue values were generally higher in microcosms, while aqueous half-life was shorter in the smaller systems.
Investigating the Ability of Pseudomonas aeruginosa pyrE Mutants to Grow and Produce Virulence Factors
Pseudomonas aeruginosa are medically important bacteria that are notorious for causing nosocomial infections. To gain more knowledge into understanding how this organism operates, it was decided to explore the pyrimidine biosynthetic pathway. Pyrimidine synthesis, being one half of the DNA structure, makes it a very important pathway to the organism’s survivability. With previous studies being done on various genes in the pathway, pyrE has not been studied to the fullest extent. To study the function of pyrE, a site directed mutagenesis was done to completely knock out pyrE, which encodes the protein orotate phosphoribosyl transferase that is responsible for converting orotate into orotate monophosphate (OMP). A mutation in this step leads to accumulation and secretion of orotate into the medium. Analyzing virulence factors produced by the mutant and comparing to the wild type, some intriguing features of the mutant were discovered. One of the findings was the over expression of virulence factors pyoverdin and pyocyanin. Pyocyanin over expression, based on the results of this study, is due to the accumulation of orotate while over production of pyoverdin is due to the accumulation of dihydroorotate. The other virulence factors studied were motility assays, exoproducts, and growth analysis. All virulence factor production was reduced significantly in the mutant compared to the wild type. The casein protease assay showed absolutely no production of proteases in the mutant. The conclusion is that orotate accumulation leads to a significant reduction in virulence factor production in Pseudomonas aeruginosa. In addition to that, it was found that excess orotate in the wild type led to a decrease in quorum sensing regulated products.
Investigation of Pyrimidine Salvage Pathways to Categorize Indigenous Soil Bacteria of Agricultural and Medical Importance and Analysis of the Pyrimidine Biosynthetic Pathway's Enzyme Properties for Correlating Cell Morphology to Function in All Phases of Growth
This dissertation comprises three parts and is presented in two chapters. Chapter 1 concerns Arthrobacter, a bacterium with an intriguing growth cycle. Whereas most bacteria exist as either a rod or coccus, this bacterium shares the rod/coccus lifestyle. It therefore seemed important to examine the growth regulatory pathways from the rod and coccus. The committed step, that catalyzed by aspartate transcarbamoylase (ATCase), in the pyrimidine biosynthetic pathway was chosen. The ATCase in Arthrobacter is like the well known Pseudomonas enzyme except that it has an active dihydroorotase (DHOase) associated. Included in Chapter 1 is the description of a microorganism, Burkholderia cepacia, whose ATCase has characteristics that are at once reminiscent of bacteria, mammals, and fungi. It differs in size or aggregation based on environmental conditions. In addition, it has an active DHOase associated with the ATCase, like Arthrobacter. B. cepacia is important both medically and for bioremediation. Since B. cepacia is resistant to most antibiotics, its unique ATCase is a prime target for inhibition. Whereas the first chapter deals with the de novo pathway to making pyrimidines, which is found mainly in the lag and log phase, Chapter 2 addresses the salvage pathway, which comes more into play during the stationary phase. This section focuses on the isolation, identification, and grouping of a number of natural soil bacteria from various soil locations. These organisms are important agriculturally, medically, and industrially. Addition of these soil isolates to poor soils has been found to improve the soil. In a previous study by D.A. Beck, the salvage schemes for a number of laboratory strains of microorganisms were determined. Nine separate classes of salvage were designated by determining the salvage enzymes present. In this study emphasis has been placed on soil bacteria, which had not previously been analyzed. A number of species of soil …
Investigations of neuronal network responses to electrical stimulation in murine spinal cultures.
Spontaneous activity in neuronal networks in vitro is common and has been well documented. However, alteration of spontaneous activity in such networks via conditioning electrical stimulation has received much less experimental attention. Two different patterns of electrical stimulation were used to enhance or depress the level of spontaneous activity in spinal cord cultures. High-frequency stimulation (HFS), a method routinely shown to increase the efficacy of synaptic transmission, was employed to augment spontaneous activity. Low-frequency stimulation (LFS), the technique often applied to depress synaptic efficacy, was employed to decrease spontaneous activity. In addition, LFS was used to reverse the effect of HFS on spontaneous activity. Likewise, HFS was applied to counter the effect of LFS. Because these networks were grown on multi-microelectrode plates (MMEPs), this allowed the simultaneous stimulation of any combination of the 64 electrodes in the array. Thus, the possible differences in response to single versus multi-electrode stimulation were also addressed. Finally, test-pulses were delivered before and after the conditioning stimulation on the same stimulation electrode(s) in order to assess the change in mean evoked action potentials (MEAPs). Dissociated spinal tissue from embryonic mice was allowed to mature into self-organized networks that exhibited spontaneous bursting activity after two weeks of incubation. Spontaneous activity was monitored from up to 14 recording channels simultaneously. Although uniform responses to stimulation across all recording electrodes were rarely observed, a large majority of the recording channels had similar responses. Spontaneous activity was increased in 52% of 89 HFS trials, whereas activity was decreased in 35% of 75 LFS trials. The duration of most of these increases was less than 5 minutes. When there were substantial and long-term (> 15 min) changes in spontaneous activity, the opposing stimulation pattern successfully reversed the effect of the previous stimulation. The percent change in MEAPs following conditioning stimulation …
The Isolation and Characterization of a Hitherto Undescribed Gram-Negative Bacterium
A unique undosciribed gramnegative rod is extensively characterized in this study. The cells of this unusual water isolate measure 1.2 X 6.5 microns, The most distinguishing characteristic of the bacterium is a polar tuft of 35-40 flagella that aggregate to function as a single organelle which is visible under phase contrast. Aging cells deposit poly- -hydroxybutyric acid granules which are bound by an inclusion membrane made up of four distinct layers. It also possesses an unusual exterior membrane outside the cell wall which contains large fibrils of protein running at a slight angle to the longitudinal axis of the cell. The guanosine-cytosine ratio was found to be 62.2$. The organism's taxonomic position was further investigated by immunological, morphological, and biochemical methods. It was found to be most closely akin to members of the genus Pseudo onas, although somewhat divergent from other species classified in this genus. After careful evaluation of the findings obtained during this study, the new bacterium was subsequently named Pseudomonas multiflagella.
Landscape Ecological Characteristics of Habitat of the Red-cockaded Woodpecker
Geographic information systems (GIS) technology was used to analyze factors influencing habitat use by an endangered species, the red-cockaded woodpecker. The study area was the western part of the Raven Ranger District of the Sam Houston National Forest, Texas. The factors considered included both structural characteristics and spatial relationships among stands of trees in the habitat.
Light and Electron Microscope Studies on the Chemotherapeutic Effect of a Combination of Dimethyl Sulfoxide and Hematoxylin on a Transplantable Lymphosarcoma
Investigations concerning the cellular response of tumor tissue to treatment with dimethyl sulfoxide and hematoxylin have not been reported. To establish the response of neoplastic tissue and cells to this combination of agents, this study was undertaken to determine the effects of dimethyl sulfoxide and hematoxylin on a transplantable lymphosarcoma in mice.
Light Spectra Distributions in Temperate Conifer-Forest Canopy Gaps, Oregon and in Tropical Cloud-Forest Canopy, Venezuela
Light spectra distributions were measured in two different montane forests: temperate and tropical. Spectral light measurements were made in different sized canopy gaps in the conifer forest at H. J. Andrews Experimental Forest in Oregon, USA. Researchers at Oregon State University created these gaps of 20 m, 30 m, and 50 m in diameter. In the tropical cloud forest, spectral light measurements were made in two plots that were permanently established at La Mucuy Parque Nacional in Venezuela, in collaboration with researchers at Universidad de Los Andes. In both studies, spectra and distributions of physiologically active light were analyzed: red, far-red, R/FR ratio, and blue light.
Odorous Metabolite and Other Selected Studies of Cyanophyta
The purpose of this study was as follows: 1. Elucidate the laboratory conditions under which Anabaena circinalis produces the noxious odor component or components in maximum concentration, 2. Isolate the major noxious odorous metabolite(s), 3. Structurally define the odorous metabolite(s), 4. Quantitate the amount of metabolite(s) that a given amount of the organism produced.
On-Road Remote Sensing of Motor Vehicle Emissions: Associations between Exhaust Pollutant Levels and Vehicle Parameters for Arizona, California, Colorado, Illinois, Texas, and Utah
On-road remote sensing has the ability to operate in real-time, and under real world conditions, making it an ideal candidate for detecting gross polluters on major freeways and thoroughfares. In this study, remote sensing was employed to detect carbon monoxide (CO), hydrocarbons (HC), and nitrogen oxide (NO). On-road remote sensing data taken from measurements performed in six states, (Arizona, California, Colorado, Illinois, Texas, and Utah) were cleaned and analyzed. Data mining and exploration were first undertaken in order to search for relationships among variables such as make, year, engine type, vehicle weight, and location. Descriptive statistics were obtained for the three pollutants of interest. The data were found to have non-normal distributions. Applied transformations were ineffective, and nonparametric tests were applied. Due to the extremely large sample size of the dataset (508,617 records), nonparametric tests resulted in "p" values that demonstrated "significance." The general linear model was selected due to its ability to handle data with non-normal distributions. The general linear model was run on each pollutant with output producing descriptive statistics, profile plots, between-subjects effects, and estimated marginal means. Due to insufficient data within certain cells, results were not obtained for gross vehicle weight and engine type. The "year" variable was not directly analyzed in the GLM because "year" was employed in a weighted least squares transformation. "Year" was found to be a source of heteroscedasticity; and therefore, the basis of a least-squares transformation. Grouped-years were analyzed using medians, and the results were displayed graphically. Based on the GLM results and descriptives, Japanese vehicles typically had the lowest CO, HC, and NO emissions, while American vehicles ranked high for the three. Illinois, ranked lowest for CO, while Texas ranked highest. Illinois and Colorado were lowest for HC emissions, while Utah and California were highest. For NO, Colorado ranked highest …
Phosphorus Retention and Fractionation in Masonry Sand and Light Weight Expanded Shale Used as Substrate in a Subsurface Flow Wetland
Constructed wetlands are considered an inefficient technology for long-term phosphorus (P) removal. The P retention effectiveness of subsurface wetlands can be improved by using appropriate substrates. The objectives of this study were to: (i) use sorption isotherms to estimate the P sorption capacity of the two materials, masonry sand and light weight expanded shale; (ii) describe dissolved P removal in small (2.7 m3) subsurface flow wetlands; (iii) quantify the forms of P retained by the substrates in the pilot cells; and (iv) use resulting data to assess the technical and economic feasibility of the most promising system to remove P. The P sorption capacity of masonry sand and expanded shale, as determined with Langmuir isotherms, was 60 mg/kg and 971 mg/kg respectively. In the pilot cells receiving secondarily treated wastewater, cells containing expanded shale retained a greater proportion of the incoming P (50.8 percent) than cells containing masonry sand (14.5 percent). After a year of operation, samples were analyzed for total P (TP) and total inorganic P (TIP). Subsamples were fractionated into labile-P, Fe+Al-bound P, humic-P, Ca+Mg-bound P, and residual-P. Means and standard deviations of TP retained by the expanded shale and masonry sand were 349 + 169 and 11.9 + 18.6 mg/kg respectively. The largest forms of P retained by the expanded shale pilot cells were Fe+Al- bound P (108 mg/kg), followed by labile-P (46.7 mg/kg) and humic-P (39.8). Increases in the P forms of masonry sand were greatest in labile-P (7.5 mg/kg). The cost of an expanded shale wetland is within the range of costs conventional technologies for P removal. Accurate cost comparisons are dependent upon expansion capacity of the system under consideration. Materials with a high P sorption capacity also have potential for enhancing P removal in other constructed wetland applications such as stormwater wetlands and wetlands …
Polycyclic-Aromatic-Hydrocarbon-Induced Alterations in the Physio-Chemical Characteristics of Escherechia Coli Dexyribonucleic Acid
Prior to 1965 the interactions of polycyclic aromatic hydrocarbons with DNA (Deoxyribonucleic acid) had been but moderately studied. It was concluded that, although a controversy existed, an apparent interaction occurred between DNA and certain aromatic hydrocarbon molecules and that the interaction was not an artifact of the reaction system.
Primary Productivity and Nutrient Relationships in Garza-Little Elm Reservoir
A large, multi-basin, reservoir (Garza-Little Elm Reservoir) in north central Texas was studied to determine the relative effects of various parameters on primary productivity. The basins were impounded several years apart,thus allowing the influence of age on water chemistry and biota to be considered. Another principal influence on water quality was secondary sewage effluent that entered one basin from a nearby source.
Remote Sensing of Landscape-Level Ecological Attributes at Ray Roberts Lake in North Texas
Biological diversity is a key component in assessing ecosystem health. Alteration, degradation and loss of habitat due to human influence is currently the primary stressor resulting in decreases in diversity. Reliable assessment of large areas in terms of biological integrity are needed for conservation and preservation efforts. Remotely sensed data provide an integrated view of reflected electromagnetic energy over large areas of the earth. These energy patterns provide unique spectral signatures which can be correlated to land cover and habitat. This research sought relationships between traditional ecological measures and information gathered from satellite digital imagery. Reliable interpretation of earth surface characteristics relies largely on accurate rectification to a map projection and subsequent thematic classification. Use of the Global Positioning System (GPS) for rectification was superior than digitizing topographical maps. Differentially corrected GPS locations provided optimum rectification with SPOT satellite imagery while marginally better rectifications were obtained for Landsat MSS imagery using uncorrected GPS positions. SPOT imagery provided more accurate land cover classifications than did MSS. Detection of temporal land cover change using MSS imagery was hampered by confusion among intermediate successional classes. Confusion between upland and bottomland forest classes occurred with both SPOT and MSS. Landscape analyses using thematic maps produced from the previously discussed endeavors suggested that terrestrial habitat in the Ray Roberts Lake area became more fragmented and complex in shape. Habitat patches became smaller but more numerous. Forested areas were most effected and conservation efforts should focus on management strategies that promote vegetation succession and forest maturation. Remotely sensed SPOT data were successfully used to predict tree basal area. There were no significant relationships found with other in situ measures or between MSS data and any vegetation measures. Remote sensing provided information suitable for large scale projects concerning landscape-level ecological issues. Rectification and classification accuracies were the …
Selected Physiological and Biochemical Studies on Blue-Green Algae
Twenty-two different unialgal clonal isolates have been obtained at random for experimental purposes over a period of about one year. Also, during this period, at least 12 other species or strains have been isolated into unialgal cultures which had not yet been identified and/or significantly cleared of heterotrophic contaminants.
Serological Characterization of Members of the Genus Streptomyces
This investigation involved isolation and characterization of specific antigenic materials from known species of Streptomyces. The resulting antigens were then used to establish serological relationships between a number of different species. These groupings were, in turn, compared with similar studies recently established by the ISP, in which other taxonomic criteria were used. By these means, the serological characteristics of the Streptomyces were evaluated for their potential utility as added taxonomic tools in the establishment of species designations within this group.
Spatially Explicit Modeling of West Nile Virus Risk Using Environmental Data
West Nile virus (WNV) is an emerging infectious disease that has widespread implications for public health practitioners across the world. Within a few years of its arrival in the United States the virus had spread across the North American continent. This research focuses on the development of a spatially explicit GIS-based predictive epidemiological model based on suitable environmental factors. We examined eleven commonly mapped environmental factors using both ordinary least squares regression (OLS) and geographically weighted regression (GWR). The GWR model was utilized to ascertain the impact of environmental factors on WNV risk patterns without the confounding effects of spatial non-stationarity that exist between place and health. It identifies the important underlying environmental factors related to suitable mosquito habitat conditions to make meaningful and spatially explicit predictions. Our model represents a multi-criteria decision analysis approach to create disease risk maps under data sparse situations. The best fitting model with an adjusted R2 of 0.71 revealed a strong association between WNV infection risk and a subset of environmental risk factors including road density, stream density, and land surface temperature. This research also postulates that understanding the underlying place characteristics and population composition for the occurrence of WNV infection is important for mitigating future outbreaks. While many spatial and aspatial models have attempted to predict the risk of WNV transmission, efforts to link these factors within a GIS framework are limited. One of the major challenges for such integration is the high dimensionality and large volumes typically associated with such models and data. This research uses a spatially explicit, multivariate geovisualization framework to integrate an environmental model of mosquito habitat with human risk factors derived from socio-economic and demographic variables. Our results show that such an integrated approach facilitates the exploratory analysis of complex data and supports reasoning about the underlying spatial …
Studies of the Passive Dispersal of Viable Algae and Protozoa by Aquatic and Terrestrial Beetles
The purpose of the present study was to determine the relative importance of aquatic and terrestrial beetles as possible vehicles in the passive overland dispersal of viable microorganisms.
Studies on Tissue Factor Pathway Inhibitor in Zebrafish
Tissue Factor Pathway Inhibitor (TFPI) is an anticoagulant protein containing three Kunitz domains, K1, K2 and K3. K1 inhibits Factor VIIa, K2 inhibits Factor Xa, and K3 enhances the Factor Xa inhibition by its interaction with Protein S. Since zebrafish is an excellent genetic model, we hypothesized that TFPI regulation could be studied using this model. As a first step, we confirmed the presence of tfpia in zebrafish. Subsequently, we performed knockdown of tfpia, and knockout of tfpia in K3 domain using CRISPR/Cas9. Both the tfpia knockdown and tfpia homozygous deletion mutants showed increased coagulation activities. Our data suggest that zebrafish tfpia is an orthologue for human TFPIα, and silencing it results in a thrombotic phenotype. We then optimized the piggyback knockdown method, where we could simultaneously piggyback 3 or 6 ASOs corresponding to 3 or 6 genes, respectively, using one VMO. These multiple gene knockdowns will increase the efficiency of genome-wide knockdowns. Since there are no studies on chromatin remodeling that control TFPI expression, we hypothesized that the genome-wide knockdowns of the Chromatin Binding and Regulatory Proteins (CBRPs) in zebrafish could help identify novel tfpia gene regulators. We chose 69 CBRPs and subjected them to simultaneous gene knockdowns. Our results have identified 5 novel regulators for tfpia. We exploited this information to discover UNC6852, a drug that enhances tfpia mRNA levels. This could be used as an antithrombotic drug. The approach developed here could be used to study the regulation of other coagulant and anticoagulant factors.
Toxicological Evaluation for the Ocular Administration of Tolrestat: An Aldose Reductase Inhibitor for the Treatment of Diabetes
Aldose reductase inhibitors (ARI's) have been shown to attenuate or prevent several complications of diabetes in animals. Tolrestat is a potent and unique ARI from Ayerst Laboratories, New York, NY. The efficacy and toxicology of tolrestat via topical ocular administration was examined. Tolrestat effectively enhanced corneal reepithelialization and was efficacious in the prevention of cataracts in the streptozotocin-diabetic rat. Ocular tissues were examined by slit lamp biomicroscopy, scanning and transmission electron microscopy, and light microscopy. Liver and kidney tissue was also examined. The presence of tolrestat was confirmed in urine, feces, and eye specimens, and quantitated in serum. There was no evidence of local or systemic tolrestat induced toxicity. Tolrestat prevented cataract formation at less than one-third the reported oral dose and at approximately one-fiftieth the associated serum concentration in rats. ED-50 and TD-50 calculations indicate that tolrestat is a relatively safe drug by topical ocular administration.
A Vegetative Analysis of and Distribution of the Grasses of North Central Texas
Accurate identification is fundamental in any study of grasses by farmers, ranchers, range specialists, ecologists, or students interested in the changes taking place in the grass habitat. Frequently it is convenient, and sometimes it is necessary, to identify the grasses by their vegetative characters. Some are readily recognized at a glance by their habitat and certain characters well known to the experienced observer. In other cases, identification is more difficult; and, perhaps with a few, it is impossible to be certain of the species from vegetative characters. However, this may also be true when the characters of the floral parts alone are considered. The inflorescence, used in most keys and descriptions, is often available only for a short period of time. Identification by the characters of the vegetation can be used throughout the growing season, even if grazing or mowing has removed or prevented the development of the floral parts. There have been other studies of grass identification related to vegetative characters, but they have been local and have not covered North Central Texas. This paper provides a means of identifying grasses by their vegetative characters. It can be used by the scientist, the technician, and the layman interested in the grasses of North Central Texas. A key using technical terminology is provided for use by the ecologist, range specialist, plant taxonomist, and student in these areas of study. For the ranchman, farmer, greens keeper, gardener, or nurseryman, a key with symbols is given that can be used without a technical knowledge of taxonomic botany. The distribution maps and diagnoses will aid any user of the keys. They will also make available additional information that may be useful in the establishment of a numerical classification and identification of plants--grasses in particular.
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