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.

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 …

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 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.

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.

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.

Quantification of stream macrobenthos populations has remained a perplexing problem in rivbrine ecology, despite numerous attempts at improvement. This is in part due to well documented variations in chemical and physical parameters locally and geographically, and resultant adapted macrobenthos populations. Southwood (1968) and Hynes (1970a) have reviewed the various sampling techniques developed'for the census of lotic macrobenthos populations. Needham and Usinger (1956), Chutter (1969), and others have pointed out the difficulty in obtaining adequate numbers of samples which will yield population estimates with desired statistical confidence, and still maintain some degree of sampling economy. Needham and Usinger (1956) and Gaufin et al. (1956) mentioned the "patchy" distribution of aquatic insect populations as the primary source of this difficulty. The concept of patchy distribution in insect populations was originally discussed by Andrewartha (1961). Attempts to improve confidence through improved sampling devices and techniques have led to development of numerous types of samplers. Cummins (1962) indicated that there were almost as many samplers as there were researchers.

Young, middle-aged and old rat adrenal gland steroidogenesis was measured in isolated, superfused glands and in their subcellular fractions before and after adrenocorticotropic hormone treatment. In the latter experiment, five corticosteroids were extracted from six different subcellular fractions. Superfused glands initially produced relatively high glucocorticoid levels; thereafter, production decayed asymptotically. Steroidogenesis by young and middle-aged glands was maintained at least 1.5 to 2.5 hours before it decayed; old glands were 50% less active than younger ones and production decayed within one hour. High cholesterol and progesterone levels in certain old gland fractions were associated with correspondingly reduced 11-deoxycorticosterone and corticosterone. It is suggested that synthesis of these glucocorticoids from their accumulating precursors weakens with age.

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.

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.

The life history of Isonychia sicca (Walsh) was elucidated from samples collected at Clear Creek from Oct. 1976-Jun. 1978, and Elm Fork of the Trinity River from Sept. 1977-Jun. 1978, Denton County, Texas. Adaptations for existence in an intermittent stream were of primary concern. Eggs are capable of diapausing through hot, dry summers and cold, wet or dry winters. Diapause is broken in the fall after rehydration and/or in the spring. I. sicca is usually bivoltine during a Sept.-Jul. wet period. Observations from Elm Fork indicate that emergence continues to Oct. if the stream remains permanent. Considerable overlap occurs between overwintering, spring, and summer populations.

A resonant frequency cavity was used to determine the dielectric properties of various preparations of Azotobacter vinelandii ATTC 12837. It was found that the bacteria investigated did interact with microwave radiation in the absence of free water. The data presented here indicate that bacteria demonstrate frequency specific dielectric properties. The techniques employed in these experiments may also be used to determine microwave spectra of other species of bacteria in different physiological stages.

Production dynamics in the phytoplankton community of a mesotrophic Texas reservoir were measured weekly over a four month period using 14C incubation and ATP assay methodologies. Assimilation values of 14C ranged from 0.2 to 45 ug C 1 hr1 - . Significant positive and occasionally negative changes in biomass (i.e. viable organic carbon) were observed in short term (4 hr) in situ incubations juxtapo-sitioned with the 14C experiments; viable organic carbon production, estimated with the ATP assay, ranged from -25 to +50 ug C 1l1hr1. Carbon assimilation and biomass changes did not correlate in either short term (4-5 hr.) or over the study period (6 months). However, weekly biomass trends were predicted by relative positive or negative biomass changes in the short term incubations. Biomass measurements gave a more sensitive insight into production dynamics in the phytoplankton community than did carbon assimilation measurements.

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.

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.

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 order to clarify the taxonomic problems which exist among the group azotobacter by serological method, 24 cultures of Azotobacter including 9 species were tested as antigens in Ouchterlony plates against 3 different antisera obtained from rabbits immunized with A. chroococcum ACl6, A. macrocytogenes St.M. and A. vinelandii 12837, respectively. The results showed more cross reactivity in intraspecies reactions than in interspecies ones and indicated the presence of a common immunogenic determinant. This work presents the results which suggest the effecient classification of Azotobacter by Ouchterlony comparisons of corresponding antigens in different species.

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,

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 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.

Rates and energies of consumption (C), egestion (F), assimilation (A), respiration (R), growth (Pg), production of exuviae (Pev), and production of egg masses (Pr) and associated efficiencies, and the effects of seasonal temperature, weight and metamorphic stage upon these factors were examined for a typical individual and cohort of Corydalus cornutus (L.) from a stream in North-Central Texas (330 23'N, 97*5'W). Dobson flies are apparently univoltine in the study area, with 11 larval instars. Emergence, oviposition and hatching occur from late May to August. The typical dobson fly hatches in mid-June, grows rapidly until November, and resumes rapid growth in March, reaching full adult size prior to leaving the stream to pupate in early June. Adult females must feed to provide energy to yolk eggs, produce egg-mass coverings and continue somatic maintenance during their week of reproductive endeavors. Metabolic compensation enables larval dobson flies to maintain preferred and fairly constant rates of R during winter (201-451 pl g-1 h~1; 5-15 C) and summer (985-1173 pl g- h1; 20-30 C); with a seasonal acclimatization change point between 15-20 C. Reduction of rates of R through undercompensation during the winter when food is scarce and through partial compensation at high temperatures during …

Glyphosate based herbicides are among the most widely used herbicides in the world. The purpose of this study was to determine developmental toxicity of glyphosate, the active ingredient in the common herbicide Roundup, on developing chicken embryos. Few studies have examined toxic effects of glyphosate alone versus the full compound formulations of Roundup, which include adjuvants and surfactants. Adjutants and surfactants are added to aid in solubility and absorption of glyphosate. In this study chicken embryos were exposed at the air cell on embryonic day 6 to 19.8 or 9.9 mg / Kg egg mass of glyphosate in Roundup or glyphosate only. Chickens treated with 19.8 and 9.9 mg / Kg glyphosate in Roundup showed significant reduction in survivability compared to glyphosate alone treatments and controls. On embryonic day 18, embryos were sacrificed for evaluation of developmental toxicity using wet embryo mass, dry embryo mass, and yolk mass as indicators. Morphology measurements were taken on liver mass, heart mass, tibiotarsus length and beak length. Embryos treated with 19.8 mg / Kg glyphosate and 9.9 mg / Kg glyphosate in Roundup showed significant reductions in wet and dry embryo mass and yolk mass. Tibiotarsus length in 9.9 mg / Kg glyphosate …

The presence of pharmaceuticals in the environment is becoming an increasing regulatory and scientific concern. Thus, the metabolic profile and bioconcentration potential of diazepam, a model benzodiazepine, were examined, as well as effects on the endocrine system in channel catfish. Through the use of specific and non-specific cytochrome P450 (CYP450) inhibitors, it was determined that CYP3A-like enzymes may play a role in the biotransformation of diazepam into temazepam; however, the isoform(s) required for the formation of other metabolites is still unknown. Overall, only around 7-8% of diazepam is biotransformed into two known metabolites. Due to the lack of inherent metabolism of diazepam in channel catfish, further analysis was conducted to determine the tissue-specific bioconcentration potential of diazepam in catfish. Various tissues were analyzed for the presence of diazepam as well as metabolites and bioconcentration factors (BCF) were calculated, which were all well below regulatory threshold values (> 2000). Additionally, modulation of the endocrine system by diazepam was examined by measuring steroid hormone concentrations and analyzing mRNA expression of selected steroidogenic enzymes and receptors. Two steroidogenic enzymes were modulated following diazepam exposure, indicating potential endocrine disrupting properties of diazepam. Together, these data suggest that diazepam exhibits low metabolic transformation rates in …

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 …