Cultured human lymphocytes may be assayed for aryl hydrocarbon hydroxylase (AHH) in whole cell preparations. The optimum assay conditions are pH 8.5, and 1.5 mM Mg++. The reaction is linear with time and cell number, and is inhibited by CO. Estradiol may inhibit induction of AHH by 3-methylcholanthrene, but is a poor competitor for the enzyme. A Caucasian population was assayed for AHH activity. The distribution was lognormal; no difference was found in cultured cells from males and females or smokers and nonsmokers. Cells from relatives of lung cancer patients showed higher activity. An American Indian population showed no difference from the Caucasian population in enzyme level. No linkage was found between AHH and 16a-hydroxylase.

An examination of the Desmodium canescens complex (D. canescens; D. tweedyi; D. illinoense) has resulted in the delimitation of a previously unreported alliance between D. canescens and D. tweedyi. The following points support this view: (a) morphological data taken from herbarium and garden specimens indicate that for many characters, the mean values of D. canescens and D. tweedy are not significantly different (b) breeding experiments have shown that artificial interspecific hybridization is possible between D. canescens and D. tweedyi (c) cytological studies have shown that D. canescens and D. tweedyi have a base number of x = 11, while D. illinoense has a base number of x = 10. A new combination is suggested: Desmodium canescens var. tweedyi (Britt.) Williams.

In view of its varied concepts and interpretations, and because of the discrepancies produced by the previous utilization of polydispersed dextrans, a study using extremely narrow-range molecular weight dextran fractions was initiated to reevaluate and consolidate some of the aspects of capillary permeability. A portion of the study was performed under decreased barometric pressure in order to clarify further some of the mechanisms involved in particulate transfer across the capillary endothelial membranes. Gel filtration procedures augmented the study as an assessment of the polydispersity effects of the dextrans employed.

The consequences of early embryonic insults and how they affect subsequent life reflects the emerging concept of "fetal programming". The aim of this project is to study the effects of embryonic insults as they subsequently manifest themselves in adults, with emphasis on the heart and vasculature. My experiments establish that fetal programming operates on the bobwhite quail inducing similar changes as those observed in mammalians and other birds. The quail's fast development provides reliable data in a short period of time than other avian models (e.g. domestic chicken). Data on quail showed a correlation between egg mass and hatchling mass; where small eggs produce small hatchlings but a high mortality made it impractical as a stressor for this study. Hypoxia was used as a stressor during embryonic incubation, where it induced a low hatching weight in quail that was not observable in adult birds. Morphological measurements demonstrated an increased ventricular collagen content and reduced ventricular lumen in birds in adults incubated in hypoxia consistent with hypertension. The hematological analyzes showed few differences indicating organ remodeling instead of hematopoietic compensation. The assessment of vascular reactivity pointed out an impaired endothelium dependent relaxation commonly associated to hypertension in birds and mammals. Fetal …

Brucella ovis is a sexually transmitted, facultatively anaerobic, intracellular bacterial pathogen of sheep (Ovis aries) and red deer (Cervus elaphus). Brucella spp. infect primarily by penetrating the mucosa and are phagocytized by host macrophages, where survival and replication occurs. At least in some species, it has been shown that entry into stationary phase is necessary for successful infection. Brucella, like other alphaproteobacteria, lack the canonical stationary phase sigma factor ?s. Research on diverse members of this large phylogenetic group indicate the widespread presence of a conserved four-gene set including an alternative ECF sigma factor, an anti-sigma factor, a response regulator (RR), and a histidine kinase (HK). The first description of the system was made in Methylobacterium extorquens where the RR, named PhyR, was found to regulate the sigma factor activity by sequestering the anti-sigma factor in a process termed "sigma factor mimicry." These systems have been associated with various types of extracellular stress responses in a number of environmental bacteria. I hypothesized that homologous genetic sequences (Bov_1604-1607), which are similarly found among all Brucella species, may regulate survival functions during pathogenesis. To further explore the involvement of this system to conditions analogous to those occurring during infection, pure cultures of …

Aspartate transcarbamoylase (ATCase) catalyzes the first committed step in the pyrimidine biosynthetic pathway. Bacterial ATCases have been divided into three classes, class A, B, and C, based on their molecular weight, holoenzyme architecture, and enzyme kinetics. Moraxella bovis is a fastidious organism, the etiologic agent of infectious bovine keratoconjunctivitis (IBK). The M. bovis ATCase was purified and characterized for the first time. It is a class A enzyme with a molecular mass of 480 to 520 kDa. It has a pH optimum of 9.5 and is stable at high temperatures. The ATCase holoenzyme is inhibited by CTP > ATP > UTP. The Km for aspartate is 1.8 mM and the Vmax 1.04 µmol per min, where the Km for carbamoylphosphate is 1.05 mM and the Vmax 1.74 µmol per min.

An aspartate transcarbamoylase (ATCase) gene from Bordetella pertussis was amplified by PCR and ligated into pT-ADV for expression in Escherichia coli. This particular ATCase (pyrB) was an inactive gene found adjacent to an inactive dihydroorotase (DHOase) gene (pyrC'). This experiment was undertaken to determine whether this pyrB gene was capable of expression alone or if it was capable of expression only when cotransformed with a functional pyrC'. When transformed into E. coli TB2 pyrB-, the gene did not produce any ATCase activity. The gene was then co-transformed into E. coli TB2 pyrB- along with a plasmid containing the pyrC' gene from Pseudomonas aeruginosa and assayed for ATCase activity. Negative results were again recorded.

The effects of thermal discharges on the density, distribution, production, and drift of benthic organisms were studied at North Lake reservoir.

Biotransformation and biodegradation rate constants were determined for naphthalene, lindane, and phenol in water samples from three different sources. Rate constants produced from monitoring disappearance of the parent chemical (biotransformation) were compared to those obtained from mineralization of the chemical (ultimate biodegradation) by ¹⁴CO₂ evolution as well as acidification of the residual ¹⁴C-labeled compound (primary biodegradation). Rate constants were statistically different for the three chemicals. The water source affected the rate constants. When biomass measurements of the waters were considered and second-order rate constants were derived, there was no statistical evidence that this parameter gave a reliable rate constant statistic that could be useful in predicting the fate of any of naphthalene, lindane, and phenol in these waters.

2,5-Diphenyloxazole (PPO) was tested as a potential alternate inducer for the aryl hydrocarbon hydroxylase (AHH) system. Its apparent lack. of carcinogenicity and toxicity provide a possible system for investigation of enzyme systems related to chemical carcinogenesis without exposure of the researcher to potent carcinogenic compounds. These studies found PPO to be an inducer of AHH in cultured human lymphocytes. When PPO was utilized as a substrate for the AHH assay system, the major metabolites produced were strongly fluorescent. A simple fluorometric assay was developed which employed PPO as the substrate and which measured constitutive activity more efficiently than similar assays using benzo(a)pyrene as the substrate. Quantitation of both basal and induced lymphocyte AHH metabolism of PPO may be applicable to human population studies and may provide a tool to determine possible genetic variables with respect to carcinogen metabolism related to cancer risk.

This study was undertaken in order to provide further insight into the effects of artificial heating on the fisheries of a small reservoir in the Southwest. The following specific objectives were established: (1) to map the reservoir for the distribution of heated water, (2) to determine the distribution of selected species of sports, rough, and forage fish in areas affected by the effluent and in areas not affected, (3) to compare size and condition of selected species of fish from areas affected by the effluent to size and condition of fish from areas not affected, and (4) to compare food habits of channel catfish in areas affected by the effluent to the food habits of channel catfish in areas not affected.

Seven species of planktonic microcrustacea were identified from the North Texas State University Golf Course Pond. Total adult microcrustacean community density, biomass and biocontent, and seasonal cycles of each species were compared with northern populations. Species diversity and evenness indices were highest in summer and lowest in spring. Variations in microcrustacean density showed a positive correlation with density of phytoplankton. Temperature had a direct effect on metabolic rates of two species of copepods. Metabolic rates of pond species were lower at common temperatures than those of northern populations. An estimate of annual energy flow through the pond ecosystem showed cladocerans contributed the greater percentage of total energy to the next trophic level.

The University of North Texas Stream Research Facility (UNTSRF) was designed to examine contaminant impacts on effluent-dominated stream ecosystems. Stream mesocosms, fed municipal effluent from the City of Denton, TX, Pecan Creek Water Reclamation Plant (PCWRP), were treated with 0, 15 or 140 µg/L cadmium for a 10-day study in August 2000. Laboratory toxicity test and stream macroinvertebrate responses indicated that cadmium bioavailability was reduced by constituents of effluent-dominated streams. The Biotic Ligand Model (BLM) for Cd was used to predict a 48 hour Cd EC50 for Ceriodaphnia dubia of 280 µg/L in these effluent-dominated streams. This value is higher that an EC50 of 38.3 µg/L Cd and a 7-day reproduction effect level of 3.3 µg/L Cd generated for C. dubia in reconstituted laboratory hard water. These results support use of a cadmium BLM for establishing site-specific acute water quality criteria in effluent-dominated streams. Although not affected by 15 µg/L treatments, organisms accumulated Cd in 15 µg/L treated streams. Hence, over longer exposure periods, Cd accumulation may increase and a no effect level may be lower than the observed 10-day no effect level of 15 µg/L. A toxicity identification evaluation procedure was utilized with in vitro and in vivo bioassays …

The effects of moderate, long-term intake of ethanol on the hypothalamic response to cold stress were examined. The long-term experimental animals were given .25 ml of 28% ethanol or .25 ml of water orally once a day, five days a week for fourteen months. A stainless steel electrode was then surgically implanted into the paraventricular nucleus, after which the animal was subjected to cold stress (-150 C, 10 min.). Recordings were taken in the forms of frequency and activity. The data clearly indicate that: (1) alcohol fed rats exhibited a suppressed response to cold stress compared to sham-fed rats; (2) this suppression of activity occurred at the level of the hypothalamus, and (3) mortality was significantly lower in alcohol-fed males than it was in sham fed males. This study clearly points out the need for further work in the area of the beneficial effects of moderate doses of alcohol.

Chick embryo fibroblasts and rat kidney cells infected in vitro with Rous Sarcoma viruses were treated with 3-deazaguanine (3-DG). The findings revealed that 3-DG inhibited virus-induced cellular transformation. Degree of inhibition is dependent on concentration and frequency of media change. 3-DG at the concentrations tested will not reverse transformed cells to the untransformed state and does not have marked effect on replication of viruses. Upon removal of 3-DG, its effect was shown to be reversible. Cell growth was generally retarded in medium containing 3-DG. When xanthosine and inosine were added to the medium, cell growth was unaffected, but it increased in guanosine.

Observations of decreases in plasma volume, shifts in plasma and lymph protein concentrations, and increases in capillary permeability at high altitude have been reported in the literature by several investigators. This investigation was begun in an attempt to elucidate the possible significance of these phenomena in future space exploration, and because of the lack of knowledge concerning the underlying mechanisms. The purpose of this investigation was to determine the effects of exposure to hypobaric pressures on the capillary permeability to the normal plasma and lymph proteins.

The effect of ethanol on the stress response in rats was examined. Experimental animals were given 0.25 ml of 28 percent ethanol or 0.25 ml of water orally once a day, five days a week, for a period of twelve months and were then subjected to fifteen minute cold stress. Corticosterone levels in ethanol-treated males following stress were significantly lower (22 percent) than in the sham group. Adrenal weights in sham-treated females were significantly higher (15 percent) than in the ethanol group at the end of twelve months. Mortality in sham-treated males was significantly higher (60 percent) than in ethanol-treated males. The effects observed may be due to the sedative action of ethanol on cortical centers controlling the hypothalmus.

N^6,O^2' -Dibutyryl cyclic adenosine 3',5'-monophosphate (Bt_2cAMP) was investigated for its effects on various tissue culture cells infected with temperature-sensitive (ts) mutant, LA31 and Bratislava 77 (B77), a wild-type Rous sarcoma virus. Specifically, known parameters of transformation were investigated and a possible site of action has been tenably proposed. The drug Bt_2cAMP was found to have little effect on the transformation related properties of primary chick embryo fibroblasts (CEF) infected with either virus or normal rat kidney fibroblasts (NRK) infected with the wild-type B77-RSV. However, significant inhibition of the transforming properties in NRK infected with the ts mutant LA31 (LA31-NRK) were reported at the permissive temperature 33 degrees centigrade (33 C).

This research represents the first systematic collection of the woody plants throughout the Cross Timbers. It provides the first keys to these plants in their vegetative condition, plant descriptions, distribution maps, and some quantitative measurements used for descriptive purposes. Descriptions of the woody plants were constructed as an aid in verification after a specimen has been identified by use of the keys. The measurements given pertain only to the woody plants as they occur in the Cross Timbers. Distributional maps are provided for all the taxa considered in this research. With the exception of those species which have the ecological amplitude to grow throughout the Cross Timbers, the distribution of the majority of the remaining species seems to be most strongly influenced by average annual precipitation. In a few instances, conditions associated with latitude appear to govern the distribution of species or varieties within the Cross Timbers. Throughout the Cross Timbers, post oak (Quercus stelta), blackjack oak (Quercus marilandica), and hickory (Caraa texan) dominate the upland forests. The streamside forests are dominated by willow (alix nigra), cottonwood (Populus deltoides), and hackberry (Celtis laevi ata). The variation in the vegetation of the Cross Timbers is not due to any change in …

Investigations have shown that the hypothalamus and pituitary respond to decreases in body temperature by stimulating thyroid release of T_3 and T_4 . This study was designed to bypass the control of the hypothalamus and pituitary gland and investigate the direct effect of temperature on the thyroid gland. Hypothermia was by an in vivo isolated perfusion of the thyroid gland. Radio-immunoassay was used to measure T_3 and T_4 concentrations. Significant increases were observed in animals perfused between 36º and 25ºc. These results indicate that the thyroid gland is directly effected by decreased temperature and that it is capable of exerting control over body temperature independent of the hypothalamus and pituitary gland. Lower perfusion temperatures produced no significant increases.

Absence of functional phenylalanine hydroxylase results in phenylketonuria (PKU). Viable treatments remain few, expensive and secondary conditions such as osteopenia occur in most PKU patients. Objective 1: Given the recently described roles of gut microbes to aid host digestion, an orally administered genetically engineered probiotic as the delivery vehicle for enzyme replacement therapy was created. The engineered probiotic, pHENOMMenal, produced phenylalanine ammonia lyase with significant production of trans-cinnamate (phenylalanine cleavage product) in vitro and resulted in a reduction of 515 μM in blood phenylalanine when fed to PKU animals for 14 days (from 2307µM ± 264µM to 1792µM ± 261µM, n = 6, P < 0.05). The control probiotic produced no change in blood phenylalanine. Thus, pHENOMMenal treatment in PKU mice demonstrated engineered microbes could compensate for a metabolic deficiency of the host. Objective 2: Evaluate the PAHenu2 mouse model of PKU for a genetic discrepancy causing ocular enlargement and delayed development observed only after the PAHenu2 mutation was crossed to the C57BL/6J mouse. When compared to healthy littermates, ELISA indicated a consistent but insignificant decrease in plasma IGF-1 and an increase in ocular IGF-1 in PKU animals. SNP screening demonstrated a differential inheritance of IGF-1 alleles in healthy and …

Crocodilians and birds comprise sister taxa of archosaurs, the development of these vertebrates occurs within an egg case that leaves developing embryos susceptible to fluctuations in the nesting environment. Studies suggest that sub-optimal conditions alter morphological growth and cardiovascular physiology. Regulation of the cardiovascular system is immature in the subjects studied, and embryos may rely on humoral rather than neural control of the cardiovascular system. The primary focus of this dissertation was to assess regulatory mechanisms responsible for maintenance of arterial pressure and heart rate. Dehydration stress had marked effects on embryo growth, and altered baseline cardiovascular parameters, while leaving the response to humoral regulator, angiotensin II (Ang II), unaffected. However, dehydrated alligator embryos developed cholinergic tone on heart rate. Hypoxic incubated chicken embryos were reduced in embryo mass, and altered response to humoral regulatory components Ang I and adenosine in addition identifying a novel regulatory component of the cardiovascular response to acute hypoxia. Collectively, these studies add to the existing knowledge of cardiovascular physiology in embryonic archosaurs and suggest that some components of cardiovascular regulation are plastic following developmental stress.

A method is presented for 160hydroxylase (SAH) induction in cultured human lymphocytes. SAH, a microsomal-associated enzyme, effects the oxidative conversion of 17pestradiol to estriol, which competes for cytoplasmic binding sites. 17,-estradiol and estrone are known mammary carcinogens, while estriol and its epimers have been suggested to have anticarcinogenic properties. To substantiate genetic variations of hydroxylase activity, an analysis of estrogen-induced cultured human lymphocytes was conducted to evaluate the frequency distribution of low, intermediate, and high SAH activity. Frequency analysis indicated that the control population distribution of SAH activity does not corroborate a proposed trimodal expansion of human SAH activity. A log normal distribution of SAH activity does exist, which suggests a polygenic mode of genetic control. SAH activity in a population of breast cancer patients and relatives of breast cancer patients showed no statistical difference from the SAH activity in the control population.

Aryl hydrocarbon hydroxylase (AHH) was induced 15-fold in Ambystoma tigrinum by intraperitoneal injection of 3-methylcholanthrene in corn oil, or 10-fold by addition of aromatic polycyclic hydrocarbons to the aqueous environment of the neotene animal. The cytochrome P-450-associated microsomal enzyme is similar to the inducible, one-gene, autosomal-dominant system typical in the laboratory mouse and man. Differences in optimal temperature for enzyme induction and activity were noted in organ culture of human and Ambystoma tissues, and ratios of benzpyrene metabolites differed between Ambystoma and Mus. The half life of enzyme activity induced in vivo was related to the excretion of hydrocarbon metabolites.