Recently, three distinct isoforms of phospholipase D (PLD) were identified in Arabidopsis thaliana. PLD α represents the well-known form found in plants, while PLD β and γ have been only recently discovered (Pappan et al., 1997b; Qin et al., 1997). These isoforms differ in substrate selectivity and cofactors required for activity. Here, I report that PLD β and γ isoforms were active toward N-acylphosphatidylethanolamine (NAPE), but PLD α was not. The ability of PLD β and γ to hydrolyze NAPE marks a key difference from PLD α. N-acylethanolamines (NAE), the hydrolytic products of NAPE by PLD β and γ, inhibited PLD α from castor bean and cabbage. Inhibition of PLD α by NAE was dose-dependent and inversely proportional to acyl chain length and degree of unsaturation. Enzyme kinetic analysis suggested non-competitive inhibition of PLD α by NAE 14:0. In addition, a 1.2-kb tobacco (Nicotiana tabacum L.) cDNA fragment was isolated that possessed a 74% amino acid identity to Arabidopsis PLD β indicating that this isoform is expressed in tobacco cells. Collectively, these results provide evidence for NAE producing PLD activities and suggest a possible regulatory role for NAE with respect to PLD α.

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.

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.

Cardiorespiratory responses to incremental dynamic exercise were assessed across four different levels of lower-body positive pressure (LBPP) and, as a separate study, during constant load (i.e constant work rate) exercise below and above each subject's ventilatory threshold (VT), both with and without 45 torr of LBPP.

Cassette systems for creating intergeneric hybrid ATCases were constructed. An MluI restriction enzyme site was introduced at the carbamoylphosphate binding site within the pyrB genes of both Pseudomonas putida and Escherichia coli. Two hybrids, E. coli pyrB polar domain fused with P. putida pyrB equatorial domain and P. putida pyrB polar domain fused with E. coli pyrB equatorial domain, are possible. The intergeneric E. coli-P. putida hybrid pyrB gene was constructed and found to encode an active ATCase which complemented an E. coli Pyr- strain. These hybrids are useful for kinetic and expression studies of ATCase in E. coli.

Cell-free extracts from Pseudomonas fluorescens NCIMB 11764 catalyzed the degradation of cyanide into products that included C02, formic acid, formamide and ammonia. Cyanide-degrading activity was localized to cytosolic cell fractions and was observed at substrate concentrations as high as 100 mM. Two cyanide degrading activities were identified by: (i) the determination of reaction products stoichiometries, (ii) requirements for NADH and oxygen, and (iii) kinetic analysis. The first activity produced CO2 and NH3 as reaction products, was dependent on oxygen and NADH for activity, and displayed an apparent Km for cyanide of 1.2 mM. The second activity generated formic acid (and NH3) pfus formamide as reaction products, was oxygen independent, and had an apparent Km of 12 mM for cyanide. The first enzymatic activity was identified as cyanide oxygenase whereas the second activity consists of two enzymes, a cyanide nitrilase (dihydratase) and putative cyanide hydratase. In addition to these enzymes, cyanide-grown cells were also induced for formate dehydrogenase (FDH), providing a means of recycling NADH utilized by cyanide oxygenase.

Asparate transcarbamoylase catalyzes the first committed step in the de novo synthesis of pyrmidine nucleotides UMP, UDP, UTP, and CTP. The archetype enzyme found in Escherichia coli (310 kDa) exhibits sigmodial substrate binding kinetics with positive control by ATP and negative control with CTP and UTP. The ATCase characterized in this study is from the extreme thermophilic Archaebacterium, Methanococcus jannaschii. The enzyme was very stable at elevated temperatures and possessed activity from 20 degrees Celsius to 90 degrees Celsius. M. Jannaschii ATCase retained 75% of its activity after incubation at 100 degrees Celsius for a period of 90 minutes. No sigmodial allosteric response to substrate for the enzyme was observed. Velocity substrate plots gave Michaelis-Menten (hyperbolic) kinetics. The Km for aspartate was 7 mM at 30 degrees Celsius and the KM for carbamoylphosphate was .125 mM. The enzyme from M. jannaschii had a broad pH response with an optimum above pH 9. Kinetic measurements were significantly affected by changes in pH and temperature. The enzyme catalyzed reaction had an energy of activation of 10,300 calories per mole. ATCase from M. jannaschii was partially purified. The enzyme was shown to have a molecular weight of 110,000 Da., with a subunit molecular …

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.

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.

Carbonic anhydrase is a ubiquitous zinc-metalloenzyme that catalyzes the interconversion of carbon dioxide and carbonate and has been found to play a wide range of roles in animals, plants and bacteria. Cotton genomic and cDNA libraries were screened for the plastidial isoform of carbonic anhydrase. The nucleotide sequences of two 1.2 Kb partial cDNA clones were determined. These clones exhibit high homology to carbonic anhydrases from other dicot plants and possess all the expected peptide motifs. For example, serine and threonine rich chloroplastic targeting peptide and conserved zinc binding residues are both present. These clones were utilized to isolate two carbonic anhydrase genes that were shown to encode different isoforms by PCR and RFLP analysis.

N-Acylphosphatidylethanoiamine (NAPE) is synthesized in the microsomes of cotton seedlings by a mechanism that is possibly unique to plants, the ATP-, Ca2+-, and CoA-independent acylation ofphosphatidylethanolamine (PE) with unesterified free fatty acids (FFAs), catalyzed by NAPE synthase. A photoreactive free fatty acid analogue, 12-[(4- azidosalicyl)amino]dodecanoic acid (ASD), and its 125I-labeled derivative acted as substrates for the NAPE synthase enzyme.

A publication by the National Academy of Sciences, USA (1979) outlined some of the research need for a great variety of economically important woody species whose remaining genetic resources need urgently to be collected and conserved. A viable regeneration system was established via tissue and cell suspension culture for Albizia falcataria and A. lebbeck, two important wood yielding leguminous tree species. The culture medium was standardized after several trials to obtain callus from the leaflet explants of these two tree species. The optimum use of casein hydrolysate (w/v) and coconut milk (v/v) in addition to 6-Benzylaminopurine and Indole-3-butyric acid could induce morphogenesis and somatic embryogenesis in the cultured tissue. This reports the first observation on somatic embryogenesis ofA. lebbeck using leaflets as the explants. Scanning Electron Microscopy and histological studies were done on the different stages plant development following standard techniques. Embryogenesis in suspension culture followed regeneration of plantlets in A. lebbeck. In A.falcaaria the regenerative process followed via organogenesis from the shoot buds developed on the leaf explants. After hardening the regenerated plants were transferred to the greenhouse. Some of the trees grew more than 25 feet tall within a few months outside the greenhouse. Karyotype of the three …

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.

The lumbar motor activity in isolated spinal cords of 72 postnatal Balb/C mice aged 2, 5, 10 and 21 days (PN2-21) was electroneurographically recorded (ENG) via bilateral ventral roots following treatment with three different concentrations (25, 100 and 200 pM) of the neurotransmitter, 5-hydroxytryptamine (5-HT), i.e., serotonin, to determine its effects on spinal pattern generation.

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.

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.

The host eosinophil response was compared in mice infected with either T. spiralis or T. pseudospiralis by determination of levels of splenic and intestinal phospholipase B, a marker enzyme for eosinophils. Primary infection of naive mice and challenge infection of homologously sensitized mice with T. pseudospiralis resulted in significantly lower tissue phospholipase B activities than infection with T. spiralis. Mice homologously challenged with T. pseudospiralis did exhibit an anamnestic eosinophil response compared to mice given a primary T. pseudospiralis infection. This anamnestic response, however, was significantly lower than the eosinophil response seen in sensitized mice given a homologous T. spiralis challenge. Mice sensitized to T. spiralis or T. pseudospiralis and heterologous challenge demonstrated an elevated eosinophil response compared to mice given a primary infection with either parasite. The heterologous challenge response, however, was not as intense as found for sensitized mice given a homologous challenge.

Changes in trophic status are often related to increases or decreases in the allocthonous inputs of nutrients from changes in land use and management practices. Lake and reservoir managers are continually faced with the questions of what to monitor, how to monitor it, and how much change is necessary to be considered significant. This study is a compilation of four manuscripts, addressing one of these questions, using data from six reservoirs in Texas.

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.

Growth of mussels under laboratory conditions was examined under various food regimes in different water types and temperatures. Growth was less than would be useful as an indicator and comparisons with field exposures were of minimal value. The effects of organophosphates on bivalves were examined via toxicity tests, tissue concentration, and by controlling exposure through the use of physical constraints. Upper temperature tolerance of both bivalve species was examined with respect to different acclimation temperatures and organophosphate exposures. Deviations from control exposures occurred at some temperatures. Copper effectively lowered the mean heat coma temperatures of C. fluminea at some concentrations, however, chlorine exposures did not alter heat coma temperature.

The physiological characteristics and organization of cat primary somatosensory cortex (SI) were studied in electrophysiological and anatomical experiments. In single cell recording experiments, quantitatively controlled mechanical stimuli were used to examine the responses of SI cortical neurons to the velocity component of skin or hair displacement. The firing frequency of most rapidly adapting neurons increased as stimulus velocity was increased. Rapidly adapting neurons were classified based on their response patterns to constant-velocity ramp stimuli. Neurons in these classes differed significantly in sensitivity to stimulus velocity and amplitude, adaptation rate, and spontaneous firing rate. The results suggest that frequency coding of stimulus displacement velocity could be performed by individual SI rapidly adapting neurons, and that the classes of rapidly adapting neurons may play different roles in sensation of tactile stimuli. Tract-tracing experiments were used to investigate the ipsilateral corticocortical connections of areas 3b and 2 in SI. Different patterns of connections were found for these areas: area 2 projects to areas 3b, 1, 3a, 5a, 4 and second somatosensory cortex (SII), and area 3b projects to areas 2, 1, 3a and SII. To further compare the organization of these areas, the thalamic input to the forepaw representation within each area was …

Habitat Evaluation Procedure data gathered at Ray Roberts Lake in 1989 and 1990 were analysed for temporal variability, observer variability and relationships between Habitat Units (HUs) and species density/diversity. observer variability within a group was analysed by cluster analysis and bootstrapping. Five out of 36 sites showed significant differences in Habitat Suitability Index (HSI) values within the group. A nonparametric Mann-Whitney test was used to analyze temporal variability. One of 6 sites showed a significant difference in HSI values between years. Using Spearman's Rank Correlation Coefficient, a correlation was found between indicator species density and HUs. No significant correlation was indicated between species diversity and HUs.

Sucrose fading was used to initiate self-administration of ethanol on an FR4 schedule in male Fischer 344 rats. Rats showed low response rates for ethanol alone. After administration of liquid diet containing ethanol, ethanol intake increased over levels prior to administration of the liquid diet. In situ hybridization compared mRNA for the inducible or constitutive 70 kD heat shock proteins in ethanol and nonethanol rats. Both inducible and constitutive mRNAs were found in nonethanol and ethanol tissues. In peripheral organs, radiolableling was higher in ethanol tissue. In brain regions, nonethanol tissues showed higher radiolabeling.

The 96-hr LC50 of cyfluthrin in Pimephales promelas ata temperature of 23*C and a pH of 8 was 1.08 g/L. The toxicity of cyfluthrin was inversely related to temperature and pH. A temperature of 10*C and a pH of 6 significantly decreased the 96-hr LC50 to 0.009 gg/L. Likewise, sublethal exposures to cyfluthrin significantly affected the fathead minnow's ability to tolerate high and low temperatures. Cyfluthrin compromised the fathead minnow's lower temperature tolerance (CTMin) by 60C and the upper temperature tolerance (CTMax) by 20C. Although cyfluthrin may not be present in the environment in large amounts due to its physical and chemical properties, small concentrations ( g/L) may adversely affect fish populations.