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.
The applicability of an isotopically labelled assay system to determine the lipase production in Pseudomonas aeruginosa was evaluated. Supernatant from cultures of Pseudomonas aeruginosa grown in a medium containing olive oil was incubated with a substrate containing labelled trioleate. Fatty acids were isolated by means of a liquid-liquid partition system. Enzyme activity was determined by measuring the amounts of free fatty acid by liquid scintillation counting. Findings indicate that the isotopicallylabelled, liquid-liquid partitioning assay is reliable, sensitive and adaptable to rapid assay conditions. It was also determined that different strains of Pseudomonas aeruginosa produce varying amounts of lipase. Partial purification of supernatant by gel filtration produced two protein peaks showing enzymatic activity.
This thesis describes the present nature of one facet of some of the flood plain forest stands in Denton County, Texas. The specific purpose was to demonstrate the presence or absence of difference between the Celtis laevigata (commonly known as the hackberry, southern hackberry or sugarberry) populations in stands on Denton Creek and Elm Fork of the Trinity River.
Starch gel electrophoresis was used to compare 14 proteins encoded by 15 loci for seven species of the family Icteridae. A close genetic relationship among these species was classified into three groups. The Agelaiine group contained Agelaius phoeniceus, Sturnella magna, and S. neglecta. The Quiscaline group contained Euphagus cyanocephalus, Cassidix mexicanus, and Quiscalus quiscula. Molothrus ater, the most divergent, was placed in a separate group. Divergence times for the seven species were compared to the literature. Heterozygosity of the seven populations of the two species of Sturnella were compared to determine factors influencing their divergence. Two factors proposed were heterosis in S. neglecta and possible hybridization between S. neglect and S. magna.
The removal of phenols from the waste waters of wood preserving plants has always presented problems. The purpose of this paper is to investigate the possibility of employing a biological system to reduce the phenol content of effluent from these plants.
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.
Liver cells from lymphosarcoma-bearing DBA/1J mice were shown, by parabiotic culture with normal liver cells from isologous mice, to elaborate an agent which could pass a 25 mu filter and transform the normal cells to a malignant state.
The peptidoglycan layer of Vitreoscilla stercoraria, ATCC 15218, was isolated from intact cells after treatment with sodium lauryl sulfate (SLS) and digestion with Pronase. Amino acid and amino sugar content was analyzed and 67% of the total present was made up of glutamic acid, alanine, diaminopimelic acid (DAP), and glucosamine in a molar ratio of 1:1.7:1:0.7. Electron microscopy of the final peptidoglycan product showed a thin, delicately folded sacculus which exhibited a morphology different from that of the intact vegetative cells. Within these sacculi occurred electron-dense structures which were assayed and found to be poly- 3-hydroxybutyrate (PHB) granules. The final yield of peptidoglycan was 2.9% of the dry weight of the intact vegetative cell.
The effect of excess gravity on bone-forming elements of the growing perichondrial shaft of embryonic chicks was investigated through the use of the transmission electron microscope and various cytochemical techniques.
This study (1) quantitatively compared steroid production in cultured Y-l mouse adrenal tumor cells exposed to Camel and Carlton-smoke derived residues, and (2) localized the effects in the cell. Basal steroid production was increased by Camel residues but not by Carlton, while ACTH stimulation was interfered with by both residues. Camel basal stimulation was comparable to that of cAMP, and was abolished by Cytochalasin D. The stimulation was also comparable to that of cholera toxin, which activates adenyl cyclase. Results indicate that residue components dissolve in the membrane stimulating adenyl cyclase at a point similar to or before that utilized by cholera toxin for its stimulating effect.
Dendrites of cockroach tibial spine mechanoreceptors contain hundreds of free microtubules, which may have some relation to the generation of electrical activity. Deflection of a spine produces a train of action potentials. Continuous perfusion over a period of 4 hours results in no response decrement. Perfusion with 10mM colchicine reversibly inhibits the response within 5-7 minutes. Irreversible inhibition is produced by perfusion with 1mM vinblastine sulfate in perfusion solution containing 1% dimethyl sulfoxide. Deuterium oxide does not inhibit at concentrations less than 50%, nor does it counteract inhibition by 10mM colchicine. Colchicine may be affecting (1) intracellular microtubules, (2) membraneous tubulin, (3) other membrane components, or (4) axoplasmic transport of essential materials to the sensory dendrites.
To better understand abiotic dynamics in Southern reservoirs receiving heated effluents, water was analyzed before and after impoundment in 330 ha North Lake. Macronutrients, metals, and chlorinated hydrocarbons were measured. Concentrations of nutrients and metals in sediments were quantified in this 2 yr study. River water prior to impoundment contained 16 times more total phosphorus, and supported 23 times more Selenastrum capricornutum cells in an algal assay than reservoir water. The reservoir has essentially no drainage and since evaporation is high, the concentrations of many dissolved solids have increased since the reservoir was filled in 1958. North Lake is now phosphorus limited. Apparently altered chemical equilibria have caused precipitation or adsorption of phosphorus with calcium and iron.
The purpose of this study was to examine the ultrastructure of both young (dividing form) and adult (stationary form) of Trypanosoma lewisi. Changes observed between the two forms of the organism were related directly to data obtained from previous comparative biochemical studies conducted on these forms of Trypanosoma lewisi.
The thermogenic effects in hypothermia of four substrates--alanine, glycine, ethano, and pyruvate - were studied in seventeen experiments. Albino rats were decapitated, and their livers were removed. The livers were homogenized with phosphate buffer at -5° C. After equilibration in a refrigerated Warburg apparatus at 20° C, the substrates were added and tissue respiration was recorded over three hours. Heat production was calculated from O2 uptake and CO2 production. Results showed that alanine, glycine, and pyrvate yielded 93.19, 89.86, and 89.89 x 10^6 kg-cal compared to a control value of 86.11 x 10^-6 kg-cal. Ethanol provided 110.31 x 10^-6 kg-cal, a value significantly greater than for the other substrates. The substrates studied, especially ethanol, did, therefore increase heat production in an artificially hypothermic environment in homogenized rat livers.
The purpose of this study was twofold. The first was to isolate, purify, and characterize the deoxyribonucleic acid (DNA) of Azotobacter vinelandii ATCO 12837. The second was to determine if there was irreversible binding of homologous 32P labeled DNA to recipient A. vinelandii cells.
This study used polyacrylamide gel electrophoresis to study the acid-phenol soluble proteins of five strains (A. vinelandii 12837, A. vinelandii 0, A. chroococcum 8004, A. macrocytogenes 8702, A. tumefaciens) of bacteria grown on Burk's nitrogen-free media, Trypticase Soy Broth, and 0.3% butanol medium. The results showed that the protein patterns can be used for the identification and possibly the taxonomic classification of the Azotobacter. The change of phenotype of the bacteria in different media followed the change of protein quantity and quality. There was no absolute similarity between any two of the species studied and this suggests a genetically heterogenous group of organisms while the amount of common proteins suggests close genetic relationships. Further studies are necessary to confirm the status of A. tumefaciens.
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.
Analysis of larval fish collected at four stations in a 330-ha cooling reservoir indicated Dorosoma spp. were most numerous at all stations, followed by Lepomis spp. and Percina Macrolepida. Largest numbers and greatest diversity of larval fish were found at the station least affected by thermal effluent; the mid-lake station provided the smallest numbers and least diversity. The two warmwater stations were intermediate, with similar numbers and diversity. Diversity and abundance of zooplankton between stations were similar to those of fish. The most abundant zooplankter (Bosmina) was generally selected against by Dorosoma, Lepomis and Micropterus spp. larvae except when the larvae were quite small ((10mm). Cyclopoid copepods were most often selected by all larvae.
The purpose of this study was to extend our understanding of the ecological relationships of P. aeruginosa by investigating the differences or similarities between the strains of this organism found in sewage and those found as pathogens in human infections. This research was approached by comparing the serological types of P. aeruginosa isolated from sewage contaminated waters in Argentina (South America) to those isolated from sewage contaminated waters in Texas. They were typed with sera obtained using P. aeruginosa isolated from human infections. The data obtained revealed that bacteria isolated from sewage in Texas and from soil and water in Argentina are antigenically similar to those isolated from human infections.
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.
Bacteria classified in the family Methylomonadaceae must derive their carbon from one-carbon compounds. They are characterized by the possession of internal membranes of two types. Type I membranes are layered and fill the middle of the cells while type II membranes form concentric layers around the periphery of the cells. Also, there are two metabolic pathways by which the methylobacteria assimilate one-carbon compounds. Further evidence of this dichotomy was sought by DNA-DNA saturation hybridization of DNAs from both types of methylobacteria. Very low DNA-DNA homology was seen between types I and II or within the types. It was not possible, therefore, to correlate the degree of genetic relatedness with either the nature of the internal membranes or the pathway of carbon assimilation.
Drumming was recorded for 11 of 13 Nearctic stonefly species, representing 4 families. Both male and female signals were obtained from 5 species, and were either 2-way or 3-way communications. Signals were species-specific; those of males and females varied from 3-39 and 1-14 beats/ signal, respectively. Duration of male signals varied from 105-8,016 ms; those of females, except Perlinella drymo (1 beat), varied from 402-1318 ms. Signals among related taxa showed greatest similarities. Duration of male signals of Perlinella drymo became progressively shorter at each of 4 temperatures from 7-29 0C. Females of Perlinella drymo would only repeatedly answer male signals recorded at near their own temperature, and would not repeatedly answer recorded male signals of 8 other species.
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.
Early function of a temperature-sensitive mutant of staphylophage 44A HJD was examined during the twenty-five-minute period following infection. Host cell and phage DNA were labeled with C and3H respectively. DNA was separated into linear and covalently closed circular (CCC) forms by density-gradient centrifugation. The host, S. aureus, shows no CCC DNA, and apparently carries no plasmid. Following infection with wild type phage, CCC DNA forms occur in tritiated and 1 C DNA fractions 10 to 15 min after infection. Infection with mutant at permissive temperature also demonstrates CCC DNA with both labels. Infection with mutant at nonpermissive temperature produced no CCC DNA during the first 25 min after infection. The impaired function in this mutant may be a linker protein.
This study dealt with the life history and population dynamics of the pine vole, Microtus pinetorum. In the past there has been a void of ecological material concerning this small mammal in the southwestern region of its range. From November, 1969, through February, 1970, a survey was conducted in Marion County and Harrison County, Texas, to determine the most suitable habitat for an ecological study of Microtus pinetorum. A study was then initiated in March, 1970, and continued through March, 1971, on an undisturbed marsh five miles south of Marshall, Harrison County, Texas.
The purpose of this study was to record the relationships between the reptiles and amphibians of Wise County and the ecological subdivisions of the area. Wise County was chosen because of the heterogenity of the area and because of its proximity to other counties which have been previously and similarly studied.
Studies were evaluated on the effects of known growth factors on the growth and carotenogenesis of Corynebacterium species strain 7ElC. The complex medium, Tryptic Soy Broth,was found to stimulate growth and production of more pigment in the light and in the dark than did a mineral salts-glucose medium. A complete amino acid mixture added to LSG enhanced carotenogenesis in the dark in Corynebacterium 7ElC, while B-vitamins retarded carotenogenesis. No absolute requirement for one or more amino acids was found,indicating a multiple amino acid requirement. The fewest amino acids found to stimulate carotenogenesis in the dark were a combination of those in the Serine and Histidine families which include serine, glycine, cysteine, and histidine.
The abundance of microtubules in receptive dendrites suggests they may function in sensory transduction. Responses of frog muscle spindle receptors and joint receptors is inhibited within 25 minutes by 50 mM colchicine, a microtubuledisrupting agent. The inhibition is reversible upon removal of colchicine, and the time course of recovery is comparable to that of inhibition. Frog olfactory responses are briefly inhibited by washing the olfactory mucosa with perfusion fluid. Colchicine accentuates the inhibition and substantially retards the rate of recovery in a dose-dependent fashion. Colchicine does not affect axonal conduction, nor the oxygen uptake of isolated crab or frog leg nerves. The inhibitory action of colchicine is therefore an effect on the electrical excitability of the receptive dendrites or soma, and not an effect on axonal conduction.
It was the purpose of this study to investigate the effect of various concentrations of IAA on the nucleic acids of Chlorella pyrenoidosa TX 7-11-05. The time during the life cycle when the greatest effect occurred was investigated by the use of synchronous cultures.
This study was designed to determine the effect of inorganic phosphate in the growth media of A. vinelandii on the synthesis and accumulation of poly-β-hydroxybutyrate (PHB) and cellular morphology. Burk's media with no added phosphate and ten times normal phosphate resulted in altered cells, lower levels of protein, and decreased cell populations. PHB accumulation was much higher in the cultures with no added phosphate; however, the polymer granules differed in appearance from those of normal cells and had a decreased rate of utilization. The amount of PHB deposited in cells grown in media containing ten times the normal phosphate level was lower than that found in regular media.
This investigation studies effects of environmental factors on growth and carotenogenesis in Corynebacterium strain 7ElC. Changes in pH were found to effect growth more than carotenogenesis. However, certain nutrients or long incubation periods stimulated carotenoid formation more than growth. Dark conditions in a mineral salts-glucose medium stimulated growth, but minimized carotenogenesis. Tryptic soy broth or yeast extract elicited carotenogenesis in darkness. Although brief light exposure during inoculation was photoinductive, continuous exposure to light following inoculation was required for maximum pigment synthesis. Dark grown stationary phase cells required 24-hours of light for maximum pigment synthesis. Chloramphenicol inhibition of carotenogenesis in dark grown cells exposed to light showed that enzymes needed for carotenoid synthesis were absent from dark grown cultures.
The effects of light, light "mimicking" chemicals, and protein synthesis inhibitors on the photo-induced carotenogenesis of Corynebacterium 7EIC were studied. Changes in the dosage of fluorescent light applied to dark grown cells showed a dose related carotenogenic response. Maintaining the same dosage but varying the wavelength of monochromatic light revealed that light with a wavelength of 280 to 450nm was responsible for photo-induction. It further showed a peak of photo-induction between the wavelengths of 370 and 430nm. The light "mimicking" chemicals antimycin A and p-Chloromercurybenzoate were shown to have no light "mimicking" effects. The transcriptional inhibitor of protein synthesis actinomycin D partially inhibited, and chloramphenicol a translational inhibitor, completely inhibited photo-induced carotenogenesis.
The role of operant behavior in the metabolism of brain 5-hydroxytryptamine (5-HT) turnover was investigated. Two and one-half hours following the administration of 150 mg/kg of para-chlorophenylalanine (PCPA), a specific inhibitor of tryptophan hydroxylase, levels of 5-HT were compared in sedentary and performing rats. Whole brain levels of serotonin were reduced in both responding and sedentary animals; however, differences between these groups were not statistically significant. The drug induced decrease in 5-HT levels was accompanied by a significant decrease in session responding. The degree of suppressed responding could be correlated with the level of 5-HT following PCPA, suggesting that the metabolism of serotonin is in part modulated by the rate of responding as maintained by the operant schedule.
This study examines ultrastructural changes occurring in seven- and fourteen-day populations of Trypanosorfa lewisi when 60 mg sodium salicylate is administered to the host. These changes were related to the host-immune response. Seven-day trypanosomes showed approximately 10 posterior volutin granules. Seven-day trypanosomes whose host had received salicylate exhibited. 30 to 40 volutin granules, and their posterior tip exhibited volutin granules in high numbers sometimes excluding other cellular elements. Fourteen-day trypanosones showed fewer volutin granules than seven-day, salicylate-treated ones. Salicylate treatment caused no additional ultrastructural alterations. Thus the volutin granules are not linked to the reproduction inhibiting antibody (ablast in) but may be involved in the formation of the trypanocidal antibodies.
The effects of various concentrations of 2-chloroethylphosphcnic acid (Ethephon), an ethylene-releasing compound, on the total protein, deoxyribonucleic acid (DNA), and ribonucleic acid (RNA) levels in Scenedesmus quadricauda IU 614 were investigated.
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.
The Cardiac Profile, a pattern of serum enzyme changes seen within seventy-two hours after an AMI, is diagnostic aid for detecting occurrence of infarcts. The effects of exercise stress on the Cardiac Profile aid clinicians in avoiding diagnostic errors in patients immediately after exercise. Five male volunteers ran from six to ten miles. Serum enzyme levels were monitored serially three days before and five days after stress. Enzyme activity was determined spectrophotometrically and electrophoretically. Significant increases in total CPK and LDH were seen. An LDH 'one-two flip' occurred eight hours after exercise. No MB-CPK was found following the run.
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 myocardium is known to release CPK, LDH1 , and GOT in response to ischemia as a result of myocardial infarction. This study was designed to induce the release of cardiac enzymes without adversely effecting the myocardium by perfusion hypothermia, thereby suggesting that these enzymes are not as specific in the diagnosis of myocardial infarction as once thought. Hypothermia was by in vivo perfusion of the left anterior descending coronary artery. Enzyme activity was measured from sera samples spectrophotometrically and electrophoretically. Significant CPK and LDH1 increases were observed in animals perfused between 25 and 19 C. These results indicate that, while heart function remained unchanged, an alteration occurred in the membrane integrity of the myocardial cells.
The work described in this thesis was undertaken to study the reasons why Azotobacter vinelandii ATCC 12837 after incubation in Burk's nitrogen-free liquid will not form as many colonies when plated on Difco Tryptic Soy Agar as when planted on Burk's nitrogen-free agar. The difference in growth of A. vinelandii on the two agars was established by performing viable cell-plate counts. The difference in growth was most apparent at 24-hours incubation of the Burk's liquid-media cultures. Phase contrast microscopic observations of Tryptic Soy media cultures of A. vinelandii disclosed the regular formation of fungoid cells at early stages of growth of the bacteria, 18 to 24 hours.
Ribavirin, a synthetic nucleoside, was found to inhibit the replication of Rous sarcoma viruses (RSV) and subsequent cell transformation in chick embryo fibroblasts (CEF). It also blocked the transformation of normal rat kidney (NRK) cells infected with temperature-sensitive mutants of RSV. The action of Ribavirin was found to be reversible as removal of the drug from the NRK cells reversed the effects on cell transformation. Ribavirin appears to have a static effect on cell growth of both NRK and CEF cells. In addition, guanosine, xanthosine and inosine altered the effect of Ribavirin on cell growth.
Laboratory studies conducted at 5, 15, 25, and 35 C measured changes in gilling rates and oxygen consumption of green sunfish in response to exposure to bentonite clay suspensions. The tests indicate that gilling rates are not affected by bentonite clay suspensions below 2125 FTU at 5 C, 1012 FTU at 15 C, and 898 FTU at 25 C. At turbidity levels exceeding 1012 FTU at 15 C and 898 at 25 C, gilling rates increased 50-70%. Tests were inconclusive at 35 C. Oxygen consumption rates were found to be unaffected by turbid suspensions below 3500 FTU at all four temperatures. Evidence suggests that increased gilling rates under highly turbid conditions are a means of compensating for reduced respiratory efficiency and a strategy for maintaining a constant oxygen uptake. Evidence indicates that the cost of increased gilling rates is probably met by a reduction in activity.
The effects of X-irradiation at dosages of 40 r, 80 r, 160 r, 240 r, 320 r, 400 r, 800 r, 4 Kr, 8 Kr, and 16 Kr on the oxygen uptake of frog brain (Rana pipiens) tissue slices were studied. A membrane-covered oxygen electrode method was used to measure the respiratory rate. Continuous recordings were made before, during, and following X-irradiation in all of the test experiments. X-irradiation was delivered from a G. E. beryllium window X-ray unit at 120 KVP, 5 ma with a 1/4 mm Al filter.
The purpose of this study was to determine the endogenous levels of indole-3-acetic acid throughout the life cycle of Chlorella pyrenoidosa, and to show a correlation between onset of cell division and IAA levels.
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