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 …

Chronic ventricular sympathectomy elicits changes in the coronary circulation, myocardial oxygen consumption and size of infarction resulting fromcoronary occlusion. These changes indicate a change occurring in the basic metabolism of the heart in response to the removal of its sympathetic nervous input. This hypothesis was tested using two groups of dogs, a shamoperated control and a ventricular sympathectomized group. The sympathectomy procedure was an intrapericardial surgical technique which selectively removes ventricular sympathetic input. Four weeks after surgery, left ventricular tissue samples were obtained and rapidly frozen to -80°C. Selected metabolic variables were then compared between the two groups.

The clcD gene encoding dienelactone hydrolase (DLH) is part of the clc gene cluster for the utilization of the B-ketoadipate pathway intermediate chlorocatechol. The roles that individual amino acids residues play in catalysis and binding of the enzyme were investigated. Using PCR a 1.9 kbp clcD fragment was amplified and subcloned yielding a 821 bp BamHi to ZscoRI subclone in the plasmid pUC19.

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.

One purpose of this investigation is to determine the fatty acid and lipid content of typical Vibrio cholerae cells. The comparison of cholera lipid constituents with those of closely-related bacteria might be of taxonomic value. Furthermore, chemical characterization of the cholera vibrio could provide useful criteria for identification of these disease-producing microorganisms.

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 …

The biosynthesis of pyrimidines in Pseudomonas putida was investigated. In this study, pyrimidine requiring mutants were isolated by conventional mutagenesis and enrichment. The strains required exogenously supplied pyrimidines for growth and were found by enzyme assays to be deficient for the product of the pyrB gene encoding the enzyme aspartate transcarbamoylase. None of the intermediates of the pathway could supply the auxotrophic requirement of the strain; only preformed pyrimidines, metabolized via salvage pathways could suffice. Pyrimidine limitation in the mutant caused a slight but significant fifty per cent increase in expression of all the de novo biosynthetic enzymes. Pyrimidine starvation's effect on nucleotide pool levels was examined in the mutant and caused a marked swelling of the purine nucleotide pools.

Stability of the macrobenthic community in a regulated section of the Brazos River, Texas, was evaluated. Physicochemical data and information on spatial distribution of macrobenthos were collected. Populations of two invertebrate species, Isonychia sicca and Neoperla clymene, had been greatly reduced since 1970-71 and I. sicca had been practically eliminated from this section of the Brazos. Three other insect populations, Choroterpes mexicanus, Cheumatopsche campyla and Cheumatopsyche lasia, had more than doubled their numbers since the 1970-71 study. A physicochemical gradient existed in this regulated section of the Brazos but it appeared to have changed little in comparison of earlier chemical data. A gradation of the macrobenthic community was evident as distance from the dam increased,

Responses of periphyton and phytoplankton productivity in the lower Sulphur River (Texas-Arkansas) to bleach-kraft mill effluent (BKME) were monitored using in situ ¹⁴C incubation. Carbon assimilation rates measured downstream of mill discharge were substantially reduced from upstream levels. Periphyton and phytoplankton chlorophyll a concentrations remained relatively unchanged by the presence of BKME. Periphyton ash-free dry weight increased near the mill outfall, but decreased further downstream. Calculated productivity efficiencies (productivity:biomass) varied with variations in ¹⁴C rates. A laboratory bioassay was designed to determine the effect of BKME light-attenuation on photosynthetic rates of upstream Sulphur River periphyton and Selenastrum capricornutum Prinz. Pooled results of bioassay runs indicated a 20 per cent BKME concentration effectively reduced control ¹⁴C-assimilation levels by 50 per cent. The downstream reduction observed for in situ productivity was 5 per cent lower than that predicted by the color bioassay.

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.

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.

Simultaneous size fractionation of plankton populations associated with NaH^14CO_3 and ^3H-glucose uptake was employed in eutrophic Lake Texoma (Texas and Oklahoma) and oligotrophic Flathead Lake (Montana). Autoradiography was utilized to determine the role of specific microorganisms in community metabolism. Ultraplankton (0.45-10 μm) dominated plankton numbers and metabolic activity in both aquatic systems. Many of the most abundant species were not the most productive, in terms of inorganic C fixation. Rates of heterotrophic uptake of ^3H-glucose were small in comparison to photolithotrophic uptake in both lakes, Photoheterotrophy was more extensive in Flathead Lake, Autoradiographs indicated that bacteria were responsible for observed photoheterotrophy. Oscillatoria sp. exhibited. mixotrophy in Lake Texoma,

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.

The feeding ecologies of leaf pack-associated systellognathan stoneflies were examined from 6 June 1980 21 May 1981. Species composition, seasonal abundance, nymphal growth, feeding habits and mouthpart morphology were determined for the eight dominant stonefly species. Prey preferences and predator-prey size relationships were also examined for omnivorous and carnivorous species. Foregut analysis from 2860 individuals indicated opportunistic feeding on the most abundant prey insects, usually in proportion to prey frequency. Feeding preference studies generally indicated random feeding on major prey groups. Prey and predator sizes were usually highly correlated (p<0.01), with predators expanding their prey size thresholds with growth. The potential for competition between sympatric stoneflies for prey is discussed.

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 …

A study was made of the effects of diphenylhydantoin (DPH) and the carrier solution on the spleen, lymph node, and thymus. DPH was injected i.p. at concentrations of 5 and 10 mg./100 gm. for 30 and 60 days. Hematologic effects observed were leucocytosis, neutrophilia, eosinophilia, and lymphopenia. Respiratory measurements of lymph node tissue slices were made using the oxygen electrode method. The carrier solution was found to cause a marked increase in oxygen consumption. A DPH effect on lymph tissue respiration was not observed. The carrier alone caused an atrophy of the lymph nodes and thymus, as well as an increase in the total body weight. Histological examination revealed that the 5 mg./100 gm. DPH injected for 60 days and the 10 mg./100 gm. DPH injected for 30 or 60 days produced a histiocytic cell type lymphoma, resembling Hodgkin's disease in the lymph node, thymus, and spleen in rats. The data indicated that DPH may not be a direct carcinogen, but it may interfere with the normal immune mechanism to produce the changes observed.

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.

Salvage of pyrimidine nucleosides and bases by S. griseus and the regulation of aspartate transcarbamoylase (ATCase) were studied. The velocity-substrate curve for S. griseus ATCase was hyperbolic for both aspartate and carbamoylphosphate. The enzyme activity was diminished in the presence of ATP, CTP, or UTP. The synthesis of ATCase was repressed in cells grown in the presence of exogenous uracil. The specific activity of cells grown with uracil was 43 percent of that for cells grown in minimal medium only. Maximal ATCase and dihydroorotase activities were found in the same column fraction after size-exclusion chromatography, suggesting that both activities could reside in the same polypeptide. The pyrimidine salvage enzymes cytosine deaminase and uridine phosphorylase were identified in S. griseus using HPLC reversed-phase chromatography.

Rates of primary production and community metabolism were monitored over a one year period using the diurnal oxygen method. Certain physico-chemical parameters were also measured, and autotrophic standing crops were estimated. An in-depth study was made of the phytoplankton community and various diversity indicies were calculated. Simple correlations were run between all parameters measured (biotic and abiotic), and their inter-relationships examined. Multiple linear regression analyses were used to develop equations predictive of production and community metabolism. Bluegreen algae were the dominant phytoplankters with blooms occurring in late summer and fall. Yearly mean production was approximately 21 kcal per meter square per day with a mean photosynthetic efficiency of 1.2 per cent. Of the various parameters measured turbidity and water temperature were most important in determining rates of primary production.

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.

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.

This research evaluated the potential of activation and attachment, as sequential companion biomarkers of phagocytosis by earthworm, Lumbricus terrestris, immunoactive coelomocytes for use in immunotoxicology. The potential was assessed by exposing earthworms to sublethal concentrations of CuSO4 and Arochlor 1254®, chemicals used as reference or standard immunotoxicants.

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.

This study concerned the effect of ethanol on cardiac activity and brain respiration in chick embryos. Ethanol dosages tested ranged from 1.0 mg to 4.0 mg/g weight. Each experiment lasted at least 150 minutes. Cardiac activity in terms of total waveform energies was integrated and printed out for plotting and analysis. The embryonic heart rates were simultaneously determined from physical graph tracings. The embryonic brain respiration was measured using a differential microrespirometer. The effect of ethanol on cardiac activity was one of slight (10 to 13), but statistically significant (p<.05) rate depression. The brain slices exhibited a marked, immediate, and irreversible decrease (39 to 89%) in oxygen consumption at both ethanol dosages. The data indicated that chick embryonic brain tissues were more susceptible to alcohol effects than cardiac tissue. Therefore, the mental abnormalities seen in the offspring of alcoholic mothers may be more cerebral in nature than cardiovascular.