This study examined central mechanisms of persistent pain using an autoradiographic technique to localize phosphoinositide hydrolysis (PI) in the rat spinal cord dorsal horn. The lateral half of laminae I-II showed the highest levels of baseline PI turnover and carbachol-stimulated PI turnover in normal animals as well as after inflammation. Inflammation resulted in increased baseline PI turnover in this region of the ipsilateral (76%) and contralateral (65%) dorsal horns. Carbachol increased PI turnover in this region in normal rats (55%) and following inflammation (ipsilateral: 46%, contralateral: 45%). The absolute magnitudes of these increases were 1.85, 2.71, and 2.51 nCi/mg, respectively. The results of this study demonstrate the involvement of PI turnover in neural mechanisms of persistent pain, and provide evidence for the involvement of cholinergic systems in this process. Because spinal cholinergic systems have been reported to be anti-nociceptive, the present results appear to reflect an upregulation of anti-nociceptive activity in response to inflammation. Thus, the spinal cholinergic system may be a regulatory site within the anti-nociceptive pathway, and may provide an attractive target for the development of new therapeutic agents.

Putative cotton cDNA clones and cognate genomic clones for a palmitoyl-acyl carrier protein (ACP) thioesterase (PATE) and an osmotin-like pathogenesis-related 5 (PR5) protein have been isolated and characterized. PATE is a class B fatty acid thioesterase with specificity for saturated long-chain fatty acids such as palmitate, and is implicated as a key enzyme to be targeted for regulation of fatty acid synthesis in order to alter cotton seed oil profiles. A nearly full-length 1.7-kb cDNA clone was isolated using a hybridization probe derived from an Arabidopsis PATE cDNA clone designated TE 3-2. A 17-kb genomic segment encompassing the PATE gene was also isolated, which has six exons and five introns with high sequence identity with other FatB cDNA/gene sequences. The deduced PATE preprotein amino acid sequence of 413 residues has putative signal sequences for targeting to the chloroplast stroma. PR5 proteins called osmotins are made in response to fungal pathogen stress or osmotic stress (water deprivation or salt exposure). Osmotins may actually form pores in fungal membranes, leading to osmotic rupture and destruction of the fungal cells. A cotton osmotin-like PR5 cDNA insert of 1,052 base-pairs was isolated and shown to encode a preprotein of 242 amino acids and is …

The pyrimidine salvage enzyme, nucleoside hydrolase, is catalyzes the irreversible hydrolysis of nucleosides into the free nucleic acid base and D-ribose. Nucleoside hydrolases have varying degrees of specificity towards purine and pyrimidine nucleosides. In E. coli, three genes were found that encode homologues of several known nucleoside hydrolases in protozoa. All three genes (designated yaaF, yeiK, and ybeK) were amplified by PCR and cloned. Two of the gene products (yeiK and ybeK) encode pyrimidine-specific nucleoside hydrolases, while the third (yaaF) encodes a nonspecific nucleoside hydrolase. All three were expressed at low levels and had different modes of regulation. As a comparative analysis, the homologous genes of Pseudomonas aeruginosa and P. fluorescens (designated nuh) were cloned. Both were determined to encode nonspecific nucleoside hydrolases. The nucleoside hydrolases of the pseudomonads exhibited markedly different modes of regulation. Both have unique promoter structures and genetic organization. Furthermore, both pseudomonad nucleoside hydrolase were found to contain an N-terminal extension of 30-35 amino acids that is shown to act as a periplasmic-signaling sequence. These are the first two nucleoside hydrolases, to date,that have been conclusively demonstrated to be exported to the periplasmic space. The physiological relevance of this is explained.

A biological monitoring tool to assess water quality using bivalve gape behavior was developed and demonstrated. The purpose of this work was to develop methodologies for screening water quality appropriate to the goals of the watershed paradigm. A model of bivalve gape behavior based on prediction of behavior using autoregressive techniques was the foundation of the bivalve biomonitoring system. Current technology was used in developing the system to provide bivalve gape state data in a continuous real-time manner. A laboratory version of the system, including data collection and analysis hardware and software, was developed for use as a toxicological assay for determination of effective concentrations of toxicant(s) or other types of stress on bivalve gape behavior. Corbicula fluminea was monitored and challenged with copper, zinc, and chlorpyrifos using the system. Effective concentrations of 176±23µg/L copper, 768±412µg/L zinc, and 68µg/L chlorpyrifos were observed using a natural water with high dissolved organic carbon concentrations. A rugged field version of the bivalve biomonitoring system was developed and deployed in two locations. The field systems were fitted with a photovoltaic array, a single board computer, and a CDPD telemetry modem for robust remote operation. Data were telemetered at a time relevant rate of once …

Roseate spoonbills are colonial nesting birds with breeding grounds extending from the United States Gulf coast to the pampas of Argentina. The U.S. population suffered a severe bottleneck from 1890 to 1920. The population's recovery was slow and partially credited to migrations from Mexican rookeries, but a gene pool reduction would be expected. Five polymorphic Spoonbill autosomal short tandem repeat (STR) loci [three (GAT)n, one (AAAG)n and one (GT)n] and one Z/W-linked microsatellite exhibiting sex-specific dimorphism were isolated and characterized. The Z/W-linked STR locus accurately confirmed the sex of each bird. Allelic profiles for 51 spoonbills obtained from Dallas (Texas), Fort Worth (Texas) and Sedgwick County (Kansas) zoos revealed a non-continuous distribution of allele frequencies, consistent with the effects of a population bottleneck. Allelic frequencies also differed significantly between the isolated zoo populations. Although extra-pair copulations were suspected and difficult to document, zoos commonly used observational studies of mating pairs to determine familial relationships among adults and offspring. STR parentage analysis of recorded family relationships excluded one or both parents in 10/25 cases studied and it was further possible to identify alternative likely parents in each case. Mistaken familial relationships quickly lead to the loss of genetic variability in captive …

Macroinvertebrate communities inhabiting an exotic, Myriophyllum spicatum, and a native, Heteranthera dubia macrophyte were studied from March 1999 to June 2000 in experimental ponds. Although macrophyte architecture explained some variation in macroinvertebrate abundance between the two macrophytes, most variation was explained by the sampling months. Total number of macroinvertebrates was found to be positively correlated with epiphyton biomass which differed significantly between the two plant types and among sampling months. Taxa richness did not vary between the two plant types. Chironomid larvae were the most abundant organisms and dominated by Apedilum elachistus on both plant communities. Annual production of five chironomid species was estimated by the size-frequency method. Production estimates (P) in g dry wt m-2 yr-1 of plant surface area for the predator Tanypodinae larvae were: Larsia decolarata, P= 0.77 and 0.67, Labrundinia virescens, P= 0.59 and 0.35 on M. spicatum and H. dubia, respectively. Larvae of Cricotopus sylvestris and Psectrocladius vernalis were collected from M. spicatum from March to mid-June. Production of C. sylvestris was found to be 0.46 g dry wt m-2, whereas it was 0.07 g dry wt m-2 for P. vernalis for this period. Apedilum elachistus exhibited the highest productivity: 9.9 g dry wt m-2 …

The objective of this research was to investigate lead speciation in the soil/sediment-water environment and to better understand how the species affect lead mobility under different environmental conditions. The research involved both field soil and sediment samples as well as standard lead solutions. Field samples were fully characterized and extracted by aqueous and organic solvents. The results were compared and evaluated with the metal speciation model, MINTEQA2. Hydrolytic polymerization and organic complexation studies were conducted with standard lead solutions under controlled experimental conditions. Results of the field samples showed that pH, dissolved cations, ionic strength, dissolved organic matter, and nature of the soil/sediment matrix play major roles in the distribution and mobility of lead (Pb) from contaminated sites. In the aqueous equilibration experiment, the magnitude of Pb2+ solubilization was in the order of pH4>pH7>pH9. The results were in good agreement with MINTEQA2 predictions. An important finding of the research is the detection of Pb polymerization species under controlled experimental conditions. At pH 5.22, Pb polymeric species were formed at rate of 0.03 per day. The role of Pb complexation with organic matter was evaluated in both field and standard samples. Different methodologies showed three types of organically bound Pb. A …

Rapid receptive field (RF) reorganization of somatosensory neurons in the rat dorsal horn was examined using extracellular single unit recording. Subcutaneous injection of lidocaine into RFs of dorsal horn neurons results in expansion of their RFs within minutes. The expanded RFs appear adjacent to or/and proximal to original RFs. Out of 63 neurons tested, 36 (58%) show RF reorganization. The data suggest that dorsal horn of spinal cord is one of the initial sites for RF reorganization. The neural mechanisms of this effect are not well understood. We propose that changes in biophysical properties (membrane conductance, length constant) of the neurons resulting from lidocaine injection contribute to RF reorganization. Iontophoretic application of glutamate onto dorsal horn neurons that show lidocaine induced RF's expansion were used to test the model. Application of glutamate produced reduction of reorganized RFs in 9 of 20 (45%) tested cells. Application of NBQX produced no effect on either original or expanded RFs indicate that RF shrinkage effects of glutamate involve NMDA receptors. The results are consistent with the prediction of the proposed model. Subcutaneous injection of capsaicin into tactile RFs of low threshold mechanoreceptive dorsal horn neurons produced no effect on the RF sizes that are …

A cotton PATE cDNA clone has a 1.7-kb insert with an coding region for 410 amino acids, lacking codons for the three N-terminal amino acids. The predicted amino acid sequence of the PATE preprotein has a characteristic stromal-targeting domain and a 63% identity to the Arabidopsis FatB1 thioesterase sequence. A cotton genomic clone containing a 17.4-kb DNA segment was found to encompass a palmitoyl-ACP thioesterase (FatB1) gene. The gene spans 3.6 kb with six exons and five introns. The six exons are identical in nucleotide sequence to the open reading frame of the corresponding cDNA, and would encode a preprotein of 413 amino acids. The preprotein is identified as a FatB thioesterase from its deduced amino acid sequence similarity to those of other FatB thioesterase preproteins. A 5'-flanking region of 914 bp was sequenced, with the potential promoter/enhancer elements including basic helix-loop-helix elements (E box). Alkaline blot hybridization of cotton genomic DNA suggests the presence at least two FatB1 thioesterase genes in cotton. Four plasmid constructs for both constitutive and seed-specific anti-sense RNA suppression and gene-transgene co- suppression of PATE gene expression were successfully generated. Two overlapping cotton genomic clones were found to encompass a Δ-12 fatty acid desaturase (FAD2-3) …

A study was conducted to evaluate the sorption characteristics of six disinfection byproducts (DBPs) on four sorbents. To investigate sorption of volatile organic compounds (VOCs), specially designed experimental batch and continuous flow modules were developed. The investigated compounds included: chloroform, 1,2-dichloroethane (DCE), trichloroethylene (TCE), bromodichloromethane (BDCM), methyl tertiary butyl ether (MTBE), bromate and bromide ions. Sorbents used included light weight aggregate (LWA), an inorganic porous material with unique surface characteristics, Amberlite® XAD-16, a weakly basic anion exchange resin, Amberjet®, a strongly basic anion exchange resin, and granular activated carbon (GAC). Batch experiments were conducted on spiked Milli-Q® and lake water matrices. Results indicate considerable sorption of TCE (68.9%), slight sorption of bromate ions (19%) and no appreciable sorption for the other test compounds on LWA. The sorption of TCE increased to 75.3% in experiments utilizing smaller LWA particle size. LWA could be a viable medium for removal of TCE from contaminated surface or groundwater sites. Amberlite® was found unsuitable for use due to its physical characteristics, and its inability to efficiently remove any of the test compounds. Amberjet® showed an excellent ability to remove the inorganic anions (>99%), and BDCM (96.9%) from aqueous solutions but with considerable elevation of pH. …

This study employed geographic information systems (GIS) technology to evaluate the vulnerability of groundwater to pesticide pollution. The study area included three provinces (namely, Kanchana Buri, Ratcha Buri, and Suphan Buri) located in the western part of central Thailand. Factors used for this purpose were soil texture, percent slope, primary land use, well depth, and monthly variance of rainfall. These factors were reclassified to a common scale showing potential to cause groundwater contamination by pesticides. This scale ranged from 5 to 1 which means high to low pollution potential. Also, each factor was assigned a weight indicating its influence on the movement of pesticides to groundwater. Well depth, the most important factor in this study, had the highest weight of 0.60 while each of the remaining factors had an equal weight of 0.10. These factors were superimposed by a method called “arithmetic overlay” to yield a composite vulnerability map of the study area. Maps showing relative vulnerability of groundwater to contamination by pesticides were produced. Each of them represented the degree of susceptibility of groundwater to be polluted by the following pesticides: 2,4-D, atrazine, carbofuran, dicofol, endosulfan, dieldrin & aldrin, endrin, heptachlor & heptachlor epoxide, total BHC, and total DDT. …

The goals of this study were to use Geographic Information Systems (GIS) and remote sensing technologies to gain a better understanding of habitat requirements of a population of ocelots in south Texas, and then apply this knowledge to form a predictive model to locate areas of suitable habitat in Willacy and Cameron counties, Texas. Satellite imagery from August 1991 and August 2000 were classified into four land cover types: closed canopy, open canopy, water, and urban/barren. These classified images were converted into digital thematic maps for use in resource utilization studies and modeling. Location estimates (762 from 1991 and 406 from 2000) were entered into a GIS in order to extract information about home range and resource selection. Each animal's home range was calculated using both Minimum Convex Polygon (MCP) and Kernel home range estimators (95% and 50%). Habitat parameters of interest were: soil, land cover, human density, road density, and distance to closest road, city and water body. Ocelots were found to prefer closed canopy and avoid open canopy land cover types. Ocelots preferred soils known to support thorn scrub, an indication of the importance of this habitat. Landscape metrics associated with habitat used by ocelots were determined through …

Pecan Creek flows southeast through the City of Denton, Texas. Characterized as an urban watershed, the basin covers approximately 63.5 km2. Pecan Creek is an intermittent stream that receives nonpoint runoff from urban landuses, and the City of Denton's wastewater treatment plant, Pecan Creek Water Reclamation Plant, discharges effluent to the stream. Downstream from the City of Denton and the wastewater treatment plant, Pecan Creek flows about 6,000 m through agricultural, pasture, and forested landscapes into Copas Cove of Lake Lewisville, creating backwater conditions. Pecan Creek water quality and chemistry were monitored from August 1997 to October 2001. Water quality was influenced by seasonal, spatial, climatic, and diurnal dynamics. Wastewater effluent discharged from the Pecan Creek Water Reclamation Plant had the greatest influence on water quality of the stream and backwaters. Water quality monitoring of Pecan Creek demonstrated that dissolved oxygen standards for the protection of aquatic life were being achieved. Water quality modeling of Pecan Creek was completed to assess future increases in effluent flow from the Pecan Creek Water Reclamation Plant. Water quality modeling indicated that dissolved oxygen standards would not be achieved at the future effluent flow of 21 MGD and at NPDES permitted loadings. Model results …