The Trinity River flows through the Dallas-Ft. Worth Metroplex in north central Texas where it receives effluents from numerous point sources including seven large regional wastewater treatment facilities. Historically, the Trinity River has been impacted by massive wastewater loadings which often constitute > 80% of the total river discharge during low flow periods. Normally, high mass loadings correspond to the summer months, compounding the effects of a naturally stressful period, characterized by high temperatures and low dissolved oxygen concentrations. Samples from 12 stations were collected quarterly over an 18 month period from the Trinity River and two tributaries. Water samples were analyzed for a variety of water quality variables, including metals, priority pollutants, pesticides, and general water quality parameters. Water samples were also tested for acute and subchronic effects with several test species. Fish were collected at each station and assemblages were characterized using traditional classification techniques and the Index of Biotic Integrity. In addition, sediment samples were assessed for toxic effects which could have adversely impacted fish recruitment and in situ biomonitoring experiments were performed. Quantitative habitat characterization analyses were performed to gain additional information that could possibly explains differences in fish assemblage structure related to habitat variability. Data …

This research project deals with the characterization of self-replicating Kirsten murine viral DNA. The replication of this viral DNA and tetracycline resistance conferred to bacteria by this viral DNA will be studied. The restriction endonuclease and Southern blot analysis revealed a fragment of pBR322 from the Hind III and Pst I site that is located in the 3' end of the MLV-K:E molecule. Single stranded sequencing of the two terminal ends of this fragment verified that the 3' end of MLV-K:E contains identical sequence homology to pBR322. The presence of this pBR322 fragment explains the unusual properties of the MLV-K:E molecule. However, tetracycline resistance is less in E. Coli containing MLV-K:E than E. coli containing pBR322 as determined by zone of inhibition assay. This may be due to alteration in the promoter region of the tetracycline gene.

The necessity for a functional user friendly laboratory data management program has become evident as the quantity of information required for modern scientific research has increased to to titanic proportions. The required union of strong computer power, ease of operation, and adaptability have until recently been outside the realm of most research laboratories. Previous systems, in addition to their high cost, are necessarily complex and require software experts in order to effect any changes that the end user might deem necessary. This study examines the Apple Macintosh computer program Hypercard as an interactive laboratory information system that is user-friendly, cost effective, and adaptable to the changing demands within a modern molecular or microbiology.

Aspartate transcarbamoylase (ATCase; encoded by pyrBI genes) is one of the most studied regulatory enzymes in bacteria. It is feedback inhibited by cytidine triphosphate (CTP) and activated by adenosine triphosphate (ATP). Much is known about the catalytic site of the enzyme, not nearly as much about the regulatory site, to which CTP binds. Until now a positive selection for feedback-modified mutants was not available. The selection we have developed involves the use of a pyrA deletion in S. typhimurium. This strain lacks carbamoylphosphate and requires both a pyrimidine and arginine for growth. In this strain citrulline is used to satisfy the pyrimidine and arginine requirements. The minimal flow through the pyrimidine pathway from the citrulline-produced carbamoylphosphate is exquisitely sensitive to feedback control of ATCase by CTP. By elevating the CTP pool, via exogenous cytidine, in a strain that also contains a cytidine deaminase mutant (cdd) growth can be stopped completely, indicating 100% inhibition. It was therefore possible to measure in vivo feedback inhibition of ATCase among the citrulline users and to isolate a family of ATCase regulatory mutants with either modified or no response to effectors.

This study had two objectives: to achieve suspension cultivation of Sf1Ep cells and to develop procedures for achieving the replication of T. pallidum in those cell cultures. Sf1Ep cells have been the sole cell line used for the in vitro cultivation of T. pallidum. A study was undertaken to determine if other cell lines can support growth of T. pallidum. Rabbit skin fibroblasts (RAB-9), nude mouse ear (NME) cells, and normal rebbit testis fibroblasts (RT) were compared to Sf1Ep cells for their ability to support in vitro multiplication of T. pallidum. RAB-9 cells supported multiplication of treponemes equal to that of Sf1Ep cells. NME and RT cells also supported growth but to a lesser extent than Sf1Ep cells. Utilization of alternative cell lines may lead to improved in vitro growth of T. pallidum including possible serial passage.

Nucleotide pools were extracted and quantified from Pyr^+ and Pyr^- strains of P. aerucjinosa. Strains were grown in succinate minimal medium with and without pyrimidines, and nucleotides were extracted using trichloracetic acid (TCA; 6% w/v). The pyrimidine requirement was satisfied by uracil, uridine, cytosine or cytidine. Pyr^- mutants were starved for pyrimidines for two hours before nucleotide levels were measured. This starvation depleted the nucleotide pools which were restored to wild type levels by the addition of pyrimidines to the medium. When the pyrimidine analogue, 6-azauracil, known to inhibit OMP decarboxylase, was added to cultures of the wild type strain, the uridine and cytidine nucleotides were depleted to near zero. Thus, the nucleotide pool levels of Pseudomonas strains can be manipulated.

Following a study of normal reproductive behavior of fathead minnows, Pimephales promelas, experiments were conducted to determine the stability/variability of behavior by sexually mature, territorial males under a variety of manipulated conditions. Collectively, these experiments indicate that although the individual behaviors of fathead minnows appear to be quite variable, the overall process, reproductive behavior, is stable.

Excessive nutrient loading of natural and artificial lakes has led, in some systems, to plethoric algal and aquatic macrophyte growth which can result in aesthetic degradation and undesirable tastes and odors. It would be advantageous to have some indication of the potential trophic status of a reservoir before it is filled. An objective of this study was to assess the water quality and nutrient loading potential of the tributaries entering Ray Roberts Lake, a large reservoir located in north central Texas. Samples from a maximum of thirteen sites were collected on the Elm Fork, Trinity River, Isle duBois Creek, and five additional tributaries. Data were also collected during six storms, from atmospheric deposition collectors, and from soil-water microcosms. The relationship between watershed landuse and mean water nutrient concentrations was evaluated. Significant differences will exist between the two major arms of Ray Roberts Lake: Elm Fork, Trinity River and Isle duBois Creek. While the majority of the annual phosphorus and nitrogen load entering both tributaries is coming from overland flow, the proportion is higher in Isle duBois Creek. Point sources in the Elm Fork contribute a larger percentage of the bioavailable phosphorus, which is significantly greater than in Isle duBois Creek. …

The effect of boron deficiency was studied in tomato plants grown hydroponically. Morphological studies showed that boron deficiency in the root system caused a cessation of root elongation, browning and thickening of root tips, the formation of branch roots close to the apex and decrease in the overall size of the root. The pyrimidine biosynthetic pathway remained intact and functional under boron limitation. The sugar nucleotide, uridine diphosphate glucose, was diminished in boron deficient medium and its level was restored to that of born sufficiency when uracil, cytosine, uridine, cytidine or orotic acid was provided.

This investigation was concerned with determining the interaction of microwaves with germinating seeds. This study covers two different approaches. The preliminary efforts covered the response of germinating seeds to treatment by microwaves and heat. The second phase of the investigation used microwaves as a probe to determine some of the processes of early seed germination. The preliminary investigation measured the internal metabolic process by ATP production. Leakage of ions and organic material from germinating seeds indicated that membranes are a target of microwaves and heat. Electron photo-micrographs showed an increase in damage to membranes as heat and microwave treatments were increased. The second phase of this investigation was concerned with determining some of the biological activity at the initiation of germination of wheat seeds, Triticum aestivum L., using a resonating microwave cavity oscillating at 9.3 GHz as a probe. Direct current conductivity measurements were also made on the seeds as a means of confirming the observations made with the microwave cavity. There was no observable difference between treatment by UHF or heat in the ultrastructure of germinating seeds. A dielectric response far above that of free water was found as live seeds of wheat began to imbibe water. This effect …

During the purification of the enzyme glycogen synthase from the muscle of the nematode Ascaris suum, approximately 70% of the glycogen synthase activity can be separated from the bulk of cellular glycogen by centrifugation for 60 min at 105,000 x . The glycogen synthase in the supernatant fraction has an Mr of 1.2 x 106 as determined by Sepharose 4B gel filtration chromatography. The glycogen synthase in this high molecular weight complex (glycogen primer complex) can be further purified by ConA-Sepharose affinity chromatography; the enzyme activity was eluted with 100 .mM a-methylmannoside. The glycogen synthase in glycogen'primer complex is predominately in the glucose 6-phosphatedependent form. The glycogen primer complex can catalyze the transfer of glucosyl units from UDP-glucose to an endogenous acceptor in the absence of exogenous glycogen. Analysis by SDS-PAGE showed three proteins (Mr 140,000, 78,000 and 34,000) and a carbohydrate polymer. The carbohydrate polymer can be partially digested with a-amylase. The glycogen primer complex was further digested by acid hydrolysis, and upon descending paper chromatography analysis, eight different carbohydrates were isolated, two of which were tentatively identified as glucose and sialic acid. The [14 C]-autoradiograph showed that in vitro synthesis of a glycogen-like polysaccharide occurred on this carbohydrate …

The objectives of this research were to identify the form of Azotobacter as it exists in situ in the soil; to compare its resistance to that of laboratory grown cysts typical of those described in the literature; and to compare its resistance to that of cells grown on dialyzed soil agar. In addition, the morphology of the cells grown on dialyzed soil agar was examined by light and electron microscopy and then compared to the cysts grown on n-butanol Burk's medium. Dipicolinic acid and oxygen uptake rate were measured in cysts and on cells grown on dialyzed soil agar in order to determine whether the cells grown on dialyzed soil agar were endospores or other dormant form and also to measure the respiratory quotient in these cells.

Respiratory and metabolic patterns in response to variations in exercise workload (WL) and pedal frequency (RPM) were examined in 10 healthy males. Each subject performed WLs of low (L), moderate (M) and high (H) intensity, equivalent to 25%, 50% and 75% V02 m a x at 7 pedal frequencies (40, 50, 60, 70, 80, 90 and 100 RPM). ANOVA ( 3 X 7 design) indicated that WL and RPM had independent and significant effects on all respiratory and metabolic measures; i.e., the greater the WL and RPM, the higher the HR, V02, VC02, Ve, Fb, Vt, Vt/Ti, Vt/Te and Ti/TtQt and the lower the Ti and Te. However, analysis of the interaction effect revealed different response patterns for Fb, Vt, Ti, Vt/Ti, Vt/Te and Ve among the WLs. During L-WL, increases in RPM produced increases in Ve which were due to progressive increases in both Fb and Vt. However, during M-WL and H-WL, increases in RPM produced increases in Ve which were accomplished by a constant Vt and a progressive increase in Fb. My findings suggest that during low WLs, the signal for Vt is dependent on rate of contraction, while during M-WL and H-WL, the signal for Vt appears …

The roles of Ca2 + and cAMP in olfactory transduction were explored using agents which affect calcium channels and second messenger systems. These agents were applied at certain calculated final concentrations onto olfactory epithelia of urethane-anesthetized frogs (Sana PiPlens) by two-sec aerosol spray. During extracellular recording, saturated vapors of isoamyl acetate were delivered every 100 sec in 0.3 sec pulses to produce an electroolfactogram (EOG). Inorganic cations that block inward calcium currents inhibit EOG responses with the following rank order: (La3+) > (Zn2+, Cd2+) > (Al3+, Ca2+, Sr2+) > (Co2+). Application of 7.5 mM La3+ eradicates £0G's, while Ba2+ (which can carry more current that Ca2+) initially produces significant enhancement (F=43.04, p<0.001, df=19). Magnesium ion has no effect on EOG's at 7.5 mM, while 1.5 X 10"4M Ca2+ is significantly inhibitory (F=5.74; p=0.0355; df=12). Control aerosol sprays of distilled water depress EOG's by an average of 5%. The organic calcium channel antagonists diltiazem and verapamil inhibit EOG's by 17% and 36X, respectively, at a concentration of 1.5 X 10~*M. Verapamil produces significant inhibition (F=17.17; p=0.002; df=ll) at 1.5 X 10" 5 M, while the 1,4-dihydropyridine calcium channel antagonists, nicardipine and nifedipine, do not inhibit beyond 1% DMSO controls. Several calmodulin …

The relationship among eosinophils, lysophospholipase activity and the immune response in animals infected with Trichinella spiralis was studied using in vivo and in vitro techniques. In an in vivo experiment, anti-thymocyte serum (ATS) was administered to mice infected with T. spiralis and its effects on intestinal lysophospholipase (EC 3.1.1.5.) activity, peripheral blood, bone marrow and intestinal eosinophilia were measured in the same experimental animal. The ATS caused a significant temporally related suppression of both the tissue lysophospholipase response and eosinophilia, in all three compartments. These findings support the hypothesis that parasite-induced eosinophilia is the cause of the increased lysophospholipase activity of parasitized tissue and that the responses are thymus cell-dependent. In vitro experiments demonstrated that the eosinophil was the primary inflammatory cell source of lysophospholipase among eosinophils, neutrophils macrophages and lymphocytes. The role of other cells and antigen in the production of the enzyme by the eosinophil was also investigated in vitro• Results demonstrated that eosinophils cultured with both T. spiralis antigen and other leukocytes yielded enzyme activities significantly greater than eosinophils cultured alone or with only antigen. More specific experiments showed that T-lymphocytes were the cells responsible for influencing the eosinophils' lysophospholipase activity in the presence of antigen, and …

Corynebacterium poinsettiae mutant strains blocked in carotenoid biosynthesis were obtained by treatment with the mutagen N-methyl-N1-nitro-N-nitrosoguanidine. Additional carotenoid (Crt) mutant strains were obtained from a previous study conducted in our laboratory. Fifty-nine Crt mutants affected in carotenoid biosynthesis were examined by a normal phase high performance liquid chromatography (HPLC) system. Pigment extracts of Crt mutants and C. poinsettiae wild type strains were resolved by an isocratic system with hexane:acetone:dicholoromethane, 11.35:1.73:1.00 (by vol.) as the eluting solvent. In addition to the five major peaks, twelve minor peaks were observed in the wild type C. poinsettiae strain used in this study. Crt mutant and wild type strain peak heights were measured from the individual chromatograms and the peak height data set created was analyzed using the Statistical Analysis System program to perform a cluster analysis. The cluster analysis revealed five carotenoid mutant groups. Carotenoid pigments which accumulated or were absent in each of the cluster groups are reported. Cluster group 1 mutants (CrtA) are blocked in the dehydrogenase(s) which is(are) responsible for the dehydrogenations between phytoene and lycopene. Cluster group 2 mutants (CrtB) appear to be blocked at a second dehydrogenase specific for the dehydrogenation from C.p. 470 to C.p. 496. Cluster …

High performance liquid chromatography was used to measure ribonucleoside triphosphates in microbial samples. Anion exchange columns in a radial compression module were used to separate and quantify purine and pyrimidine ribonucleotides. Endogenous ribonucleoside triphosphates were extracted from Escherichia coli and pseudomonas aeruginosa using three different solvents, namely trifluorocetic acid (TFA; 0.5M), trichloroacetic acid (TCA; 6 per cent w/v) and formic acid (1.0M) Extracts were assayed for uridine 5'-triphosphate (ATP), and guanosine 5'-triphosphate (GTP) by using anion exchange radial compression high performance (pressure) liquid chromatography. The three extraction produres were compared for yield of triphosphates. E. coli, the TFA extraction procedure was more sensitive and reliable than TCA and formic acid extraction procedures, but , in P. aeruginosa, the best yields of ATP and GTP were obrained following extraction with TFA. Yields of UTP and CTP increased when extraction was performed in TCA. These data illustrate that different extraction produres produce different measures for different triphosphates, a point often overlooked.

The role of Ca++ in olfactory responses was investigated with inorganic and organic calcium channel antagonists. Electrophysiological responses to odorants were recorded from frog olfactory mucosa before and after aerosol application of different agents. Electroolfactogram responses were blocked by certain inorganic ions with the order of effectiveness Zn++ >Ln+++>Cd++>Ca++>Co++>Sr++>Mg++. Ba++ potentiated olfactory responses, and is known to potentiate calcium channel-mediated responses in other tissues. Certain local anesthetics which are thought to act through calcium channel blockade were inhibitory to olfactory responses, with the order of effectiveness being dibucaine>tetracaine>procaine. These data support the idea that Ca++ is involved in olfaction, perhaps acting as a current carrier and/or a second messenger. Preliminary experiments on channel localization were performed using a silicon-labeled amine. Attempts to localize the silicon label were inconclusive, although silicon was detected in the olfactory tissue.

In recent years there has been a great deal of scientific interest in processes that affect the fate of organic chemicals in the environment. one main reason for this increased interest is due to greater environmental concern over accidental or purposeful release of these chemicals into the environment by man. A major environmental concern is the increased use of pesticides over the last few years. In the thirty years prior to 1978 the use of pesticides has increased by a factor of forty (Ridgeway et al., 1978). Recently the use of herbicides has been increasing, but that of insecticides has stabilized (Willis, 1983). Detectable amounts of organic pesticides can be found in many areas of the biosphere. For toxic organic chemicals to be used safely, researchers must have a clear understanding of the fate and persistence of these chemicals when they are released into the environment. This understanding will also allow the development of new products that, when properly used, will not produce adverse effects to man or the environment (Weber, 1972). According to the Toxic Substance Control Act (TSCA) any new or expanded-use chemical that might be released into the environment must be tested for environmental hazard.

Extraction and quantitation of myosin light chain two coupled with myograph recordings from Ascaris muscle perfused with calmodulin inhibitors and neurotransmitters in conjunction with their respective agonists and antagonists have been used to establish the regulation of contraction in this muscle. Densitometric tracings of isolectric focusing gels separating the regulatory light chain were used to quantitate phosphorylation in resting, contracted and flaccid muscle. These studies indicated that inhibitory neurostimulation is mediated by a true GABA receptor. Myosin-mediated contraction is responsible for maintaining the level of tension observed in resting actin-mediated muscle. Actin-mediated contraction is responsible for the rapid rise in tension following excitatory stimuli. Both systems function simultaneously and are independant.

Adult stonefly emergence, nymphal growth, drift and microdistribution were examined in Battle Branch, a secondorder, spring-fed stream, from November, 1982 to May, 1984. Adults of 22 species emerged successionally, with specific adults present every month. Searching emergent objects and the stream margin was best for collection of winter and early spring emerging species. Sweeping the streamside and light trapping were most efficient for late spring and summer emerging species. Nymphal growth for nine abundant species generally fit double log or semilog models. Drift was low, but did show a post-sunset pulse. Generally, leaf material was found to be significantly related (p<O.001) to density, diversity and biomass of stoneflies in Battle Branch.

Etheostoma spectabile, E. punctulatum, and Cottus carolinae were sampled Mar., 1983, - Feb., 1984, in Flint Creek, Oklahoma. Immature E. spectabile ate primarily microcrustaceans, while mature fishes relied more on mayflies and amphipods. Juvenile E. punctulatum fed upon mayflies, amphipods, and Asellus. Mature E. punctulatum ate primarily mayflies, and other relatively larger prey. Cottus carolinae consumed chironomids almost exclusively in Jan. - Feb., 1984, while mayflies were predominant the remainder of the year. No significant habitat partitioning between the two darters, and seasonal habitat segregation between C. carolinae and the two darters was found. Dietary overlap between the darters was significantly correlated (p<0.0005) to differences in x prey sizes.

A comparative study was made of environmental variables and the density, biomass, diversity, and species composition of macroinvertebrates in areas downstream from a dam with a hypolimnetic release (Hungry Horse Dam on the Flathead River) and a dam with a selective withdrawal system (Libby Dam on the Kootenai River). A major objective of this study was to examine the response of macroinvertebrate communities to defined environmental gradients in temperature, flow, substrate, and food-related variables (periphyton, particulate organic carbon in the seston). In addition, the effects of experimental manipulations in discharge on macroinvertebrate drift and stranding were assessed, and the effects of temperature on the growth rates and emergence of five species of insects were measured.

Twenty-two natural populations of the red shiner minnow, Notropis lutrensis were examined for morphological and genetic variation. This research was aimed at testing the hypothesis that morphological and genetic variation was primarily influenced by the degree of gene flow between populations. Ten linear measurements were taken from each of 1320 specimens. Morphological characters were adjusted for differential growth by least squares linear regression techniques. Genetic variability was estimated for each individual red shiner through the methods of starch gel electrophoresis. Twenty presumtive gene loci were resolved.