Biochemistry, Biophysics, and Structural Biology Commons^™

Effects Of Glucocorticoid Treatment On The Properties Of Single Notor Units In Cat Hindlinb Huscles, Andrew James Robinson

Theses and Dissertations

The administration of glucocorticoids has been shown to induce atrophy and weakness in whole skeletal muscle. These effects frequently appear to be more pronounced in pale muscle than in red muscle. The present study was undertaken to examine the effects of steroid treatment on the contractile, electrical, and fatigue properties of single meter units in pale and red cat hindlimb muscles. Single motor units in the medial gastrocnemius (MG) and soleus muscles were isolated using techniques developed by McPhedran, et al., (1965). The properties of a large number of motor units were examined in normal animals and were compared to …

The Relationship Of Reduced Levels Of Atp To Inhibited Chondrogenesis In The Production Of Limb Deformity Induced By 6-Aminonicotinamide, Val C. Sheffield

Theses and Dissertations

6-aminonicotinamide (6-AN), an analogue and antagonist of nicotinamide, impairs cartilage formation and results in shortening of the limb when administered to chick embryos. 6-AN forms an abnormal NAD analogue (6-ANAD), which inhibits the activity of NAD-dependent enzymes associated with the production of ATP. The mechanism of action of 6-AN was studied by measuring the biosynthesis of protein and DNA, and the sulfation and glycosylation of chondroitin sulfate in control and 6-AN treated cartilage from 8-day chick embryos. The cartilage was also assayed for ATP levels. Incorporation of Na235SO4 was inhibited by 6 h treatment with 10 μg/ml 6-AN, whereas 3H-thymidine …

Isolation, Purification, Scanning Electron Microscopy And Bacterial Dna Uptake Of Plant Protoplasts, Bronwyn G. Hughes

Theses and Dissertations

Protoplasts were isolated from tobacco and barley leaves in sucrose or mannitol using commercially available cellulases and macerozymes. Barley growth and protoplast isolation and purification conditions were optimized so that protoplasts were obtained in high yields free of unwanted debris and organelles. A technique for processing barley and tobacco protoplasts for examination by scanning electron microscopy was developed in which protoplasts seem to have maintained their structural integrity. Barley and tobacco protoplasts took up 3H-B. subtilis DNA, 125I-B. subtilis DNA or 125I-M. luteus DNA as a linear function of time (0-6 hr) and DNA concentration (0-200 μg/ml). Up to 16 …

Direct Chemical Evidence For A Supramolecular Protein Structure In The Nuclear Envelope, David Lee Cochran

Theses and Dissertations

Protein subunit structure in the nuclear envelope has not been previously described. Covalent crosslinking of polypeptides provides a method for studying the subunit structure of protein-rich systems. In this study, several polypeptide crosslinking methods have been used to determine associations among polypeptides in isolated chicken erythrocyte nuclear envelope. Interchain associations that exist in the isolated membrane were fixed by crosslinking the polypeptides in the isolated nuclear envelope. Crosslinked and uncrosslinked polypeptides were resolved by electrophoresis in a dissociating detergent system. Each agent caused distinctive alterations in the gel pattern. Certain bands diminished and disappeared while new bands of two or …

Effect Of Altitude On The Peripheral Degradation Of Thyroxine In The Uinta Ground Squirrel (Citellus Armatus), Stuart K. Ware

Theses and Dissertations

Various methods were employed to detect changes in the peripheral degradation of thyroxine (T4) in adult Uinta ground squirrels (Citellus armatus) collected at two different altitudes (5400 and 9000 feet.) Plasma half-life of injected T4-125I (tl/2) was significantly decreased with altitude, as was T4-125I distribution space (TDS) and T4 degradation and secretion rate (TSR). In vitro deiodination of T4-125I by heart homogenates was not affected, while urinary excretion of thyroxine derived iodide was increased at high altitude 44-70 hours post-injection. Both total and free serum T4 concentrations were reduced at high altitude, although not significantly. The cause of the reduced …

A Spectrophotofluorometric Analysis Of Serum Vitamin A Levels In A Defined Population, James Ronald Schroeder

Theses and Dissertations

The objectives of this thesis were to examine serum vitamin A levels in two defined populations. The method of analysis was a spectrophotofluorometric technique with a correction formula to account for interfering fluorescence from an identified carotenoid phytofluene (37).

The first population were infants at time of delivery. Infants were divided into two groups depending upon length of gestation. A premature group was identified as having a length of gestation of less than 36 weeks. Term infants were defined as 36 weeks or greater. The mean value of serum vitamin A of eight premature infants was 10.5 μg/100ml (S.E.M.+.78). The …

Studies Of N⁵, N¹⁰-Methylene Tetrahydrofolate Reductase From Porcine Kidney And Mouse L1210-Induced Tumor Tissues, Purification And Interaction With Antifolates, David W. Jayme

Theses and Dissertations

Methylene tetrahydrofolate reductase has been purified 1000-fold from porcine kidney and 400-fold from mouse L1210-induced tumor tissue by classical methods. The enzyme preparations have been demonstrated to be essentially free of contaminating methionine synthetase and serine transhydroxymethylase activity. Studies of the kinetic properties of the kidney and tumor enzymes, with respect to the reverse reaction using N5-methyl tetrahydrofolate as the variable substrate, have indicated Km values of 2.0 and 2.4 x 10-4 M, respectively. Inhibition of this key branch point enzyme in folate metabolism by a number of antimetabolites indicates that several of these antifolate compounds exhibit enzyme inhibition superior …

The Effect Of Altitude On Thyroid Function Of The Uinta Ground Squirrel (Citellus Armatus), Wendell Lee Wells

Theses and Dissertations

Thyroid uptake and distribution of 125I, concentrations of T4 and T3 in serum and thyroids and concentrations of TSH in serum and pituitaries were measured to determine thyroid function of Uinta Ground Squirrels (Citellus armatus) native to 5,400 and 9,000 feet elevations to evaluate alterations in the state of the thyroid due to increased altitude. Uptake of 125I was 60 percent lower by squirrels from 9,000 feet. Incorporation of 125I into T3 was greater at the higher elevation, but absolute thyroid T3 concentrations were similar at both elevations. No significant differences were found in serum T3 concentrations, but squirrels living …

Absorption Of Caffeine Through Isolated Rat Small Intestine, William James Hatch

Theses and Dissertations

Intestines of fifty-six albino rats of the Holtzman strain were removed and perfused with Krebs bicarbonate buffered solutions containing caffeine. After one hour of absorbtion, caffeine was extracted from the serosal fluids in chloroform and concentrations were determined spectrophotometrically. It was found that all portions of the small intestine transported caffeine against a concentration gradient. This transport mechanism demonstrated saturation kinetics and was inhibited by ouabain. It was concluded that the movement of caffeine from the mucosa to the serosa of the small intestine cannot be the result of simple diffusion. Possible mechanisms by which the absorption of caffeine takes …

Hepatic Ornithine Metabolism In Hydrazine-Treated Rats, Van Saxton Hubbard

Theses and Dissertations

Ornithine plays an important role in mammalian intermediary metabolism. Ornithine can be (1) utilized as a component of the urea cycle, (2) decarboxylated to form putrescine, a precursor of the polyamines, spermidine and spermine, and (3) converted metabolically to arginine, proline and glutamate. In prior investigations conducted in our laboratories and others, free endogenous hepatic pools of ornithine were found to be elevated following hydrazine administration. Consequently, time-course alterations of arginase, ornithine δ-transaminase, ornithine transcarbamylase, and ornithine decarboxylase activities were investigated regarding the effect of hydrazine treatment on hepatic ornithine metabolism in the rat. As an outgrowth of these investigations, …

I. An Isothermal Titration Microcalorimeter. Ii. Importance Of Micronutrients In Nutrition. Iii. Zinc Bibliography, John Willard Gardner

Theses and Dissertations

I. An isothermal titration microcalorimeter having a colume of 4 ml and capable of temerature control to ±2 x 10-5°C is described. Major components include a constant temperature water bath controlled to ±3 x 10-4°C, a platinum reation vessel, and an isothermal control circuit consisting of constant Peltier thermoelectric cooling and variable Joule heating controlled by a thermistor in an AC Wheatstone bridge circuit. The calorimeter was tested by measuring the heat of ionization of water and was found to produce data accurate to ± 0.1% where small samples are used such as in the investigation of many biological systems.

The Effects Of Deficiencies Of Certain Mineral Nutrients On The Activity And Various Properties Of Ribulose-1,5-Diphosphate Carboxylase From Barley (Hordeum Vulgare L.), Philip Alan Harrison

Theses and Dissertations

Barley plants grown in Mg, Mn, Zn and Cu deficient media were analyzed for specific activity, total activity and amount of RuDPCase present. Percent dry weight, as well as chlorophyll and soluble protein content were determined. All deficiencies caused lower chlorophyll and RuDPCase concentrations than occurred in the control. Soluble protein content, however, was higher in all deficient plants than in the control, either on a dry or a fresh weight basis. The addition of 18 μM Mn^2+ increased the activity of Mg^2+ containing RuDPCase preparations as much as 133% above normal. The specific activity of RuDPCase from Mg, Mn, …

The Effects Of Thiamin Deficiency And Thiamin Antagonists On Cardiac Function In The Rat, D. James B. Sutherland

Theses and Dissertations

A study was made to determine what biochemical changes were involved in the slowing of the rat heart rate during thiamin deprivation, pyrithiamin or oxythiamin treatment. The appearance of bradycardia during progressive thiamin deficiency was shown to be related to the reduction in myocardial enzymatic activity of pyruvate (PDH) and 2-ketoglutarate (2-KGDH) dehydrogenase, and not due to inanition alone since pair-fed control rats did not show bradycardia until the agonal stage. Bradycardia was not the result of reduced energy since CP, ATP, ADP, and AMP levels remained normal even though PDH and 2-KGDH activity dropped to 20% of normal by …

Transforming Ability Of Bacillus Subtilis Dna Taken Up By Barley Embryos, Christine Tolman Ence

Theses and Dissertations

Purified Bacillus subtilis DNA taken up by one-day-old barley embryos and permanently fixed within the barley cell is shown to have maintained its biological activity in B. subtilis transformation. Cesuim chloride density gradient centrifugation of DNA isolated from the roots of barley embryos after the uptake and incorporation of radioactive, bromouracil-labelled B. subtilis DNA indicates the presence of the bacterial DNA in several discreet bands of well-defined density, whose density is changed by sonication. These preliminary findings concerning the fate of bacterial DNA taken up by barley roots are discussed in terms of the recombination model of Ledoux and Huart …

Lipids Of Fungal Spores, Identification And Metabolism, Jim L. Bushnell

Theses and Dissertations

The lipids of the spores of the Basidiomycetes: Ustilago bullata, Lycoperdon perlatum and Calvatia gigantea were investigated. The spores of U. bullata were found to have hydrocarbons ranging from C_9 to C_24, methyl esters of fatty acids ranging from C_14 to C_24, and free fatty acids from C_14 to C_24. The spores of L. perlatum were found to have few hydrocarbons (C_22 being the predominent one); no methyl esters were detected and the fatty acids were from C_15 to C_18. Long chain aldehydes (C_30) were also detected in low concentration. At the ultrastructural level the Lycoperdon spores were found to …

Enzymatic Synthesis Of Oligoribonucleotides Of Defined Base Sequence, Bronwyn Geraldine Hughes

Theses and Dissertations

A possible method for the synthesis of ordered oligoribonucleotides involves primer-dependent polynucleotide phosphorylase (PNPase) synthesis using the RNase A or T1 resistant N-cyclohexyl-N'-β(4-methylmorpholinium)ethylcarbodiimide chloride (CMC-Cl) derivatives of uridine or guanosine containing primers. Thus, CMC-UpA, CMC-GpA, CMC-UpCpC, CMC-GpApC, and ApApApApU-CMC were prepared and studied as primers for PNPase in 15 min 14C-ADP polymerization reactions and also in 6-10 hr synthesis reactions. The CMC-primers were not suitable primers for PNPase catalyzed synthesis reactions for either time. The PNPases used in such studies were contaminated with nuclease that showed the following order of susceptibility: UpA > CpC, GpA > ApU, ApA. Even a highly purified …

Autoradiographic Studies Of The Distribution Of Serotonin In The Rat Brain, Brent D. Wagstaff

Theses and Dissertations

Serotonin (5-hydroxytryptamine, 5-HT), considered to be a synaptic transmitter within the central nervous system, has been shown to be localized mainly in the paleocortex of the brain. Others have shown disturbances of normal 5-HT metabolism to be associated with mental illness and disease. Attempts to inject tritium labelled 5-HT into cerebrospinal fluid of the rat brain for anatomical localization studies were·made. The following observations were noted, (1) there exists a perivascular space in which fluid flows and distribution is relatively uninhibited as compared to subarachnoid space, (2) the labelled 5-HT penetration of brain tissue parenchyma is restricted and radioactivity is …

Effect Of Exercise On Tissue Concentrations Of Thyroid Hormones And On Thyroxine Degradation Rate In The Rat, William W. Winder

Theses and Dissertations

Several methods were used to detect changes in thyroid activity and in degradation of thyroxine (T4) in exercising and sedentary control rats. Rats were run in a motor-driven exercise wheel over a three-week period at progressively increasing work loads and then were run 1.5-2 hours per day for 10-14 days before sacrifice. Disappearance of injected T4-125I from plasma was considerably enhanced by exercise. As determined by isotopic equilibration and paper chromatographic methods, T4 concentration was depressed in plasma and elevated in liver of exercised rats. No significant changes in T4 concentration were observed in heart, kidney, or gastrocnemius muscle. Thyroidal …

The Biochemical Response Of Provo Bay To Nutrient Inflow, R. Bruce Sundrud

Theses and Dissertations

Provo Bay of Utah Lake, Utah, receives the effluents from farms, industry, and three cities. In order to determine the effects of these effluents, eleven stations were established throughout the Bay. At weekly intervals from June 19 to October 26, 1970, and monthly thereafter until March, 1971, the water at these stations was sampled for dissolved oxygen (DO}, carbon dioxide (CO2), turbidity, pH, phosphates, nitrates, biochemical oxygen demand (BOD) and coliform bacteria. Due to intense algal blooms, the quality of the water changes as it passes through Provo Bay. Average values for the inflow, mid-Bay, and point of discharge respectively …

The Effects Of 2-Chloro-4-Ethylamino-6-Isopropylamino-S-Triazine (Atrazine) On The Activities Of Certain Calvin Cycle Enzymes In Barley And Corn, L. Chris Harrison

Theses and Dissertations

Atrazine (2-chloro-4, ethylamino-6-isopropylaminos- triazine) applied to Hordium vulgare L. variety barbless in low concentrations (0.001, 0.01, 0.1 uM) was shown to in--­ crease the activity of D-ribulose-1, 5-diphosphate carboxylase (RuDPCase) and phosphoribulokinase (PRuK) up to 23% and 30% respectively and increase the amount of chlorophyll and extractable protein up to 18% and 21% respectively over the control plants. These results indicate that sub-toxic levels of atrazine significantly stimulate plant growth. Barley plants given higher concentrations of atrazine (1.0 and 10 uM) showed up to a 78% and 42% decrease in the activity of RuDPCase and PRuK respectively as well as …

Evaluation Of The Essentiality Of Dextran In The Dental Caries Process, William Redman Grigsby

Theses and Dissertations

In summary, the main points to support the view that dextran synthesis is unessential to the dental caries process are presented below:

1 . Dextran-fed, microbe-bearing rats do not experience dental caries.

2 . Lactobacillus casei 4646 has no extracellular dextransucrase enzyme or other extracellular enzymes which release glucose or fructose from sucrose .

3. L. casei 4646 does not synthesize extracellular polysaccharide of the dextran class when grown on sucrose-containing medium.

4. L. casei 4646 does not exhibit in vitro, adherent growth on stainless-steel wire when grown on sucrose-containing medium .

5. L. casei 4646 does not cause dental …

A Study Of Folic Acid Dependent Reactions In Bovine Brain, Dillard Spencer Broderick

Theses and Dissertations

The enzymatic interconversion of serine to glycine and N5, N10-methylene tetrahydrofolate to N5 -methyl tetrahydrofolate have been studied in a cell-free system derived from bovine brain. A partial purification of serine transhydroxymethylase and N5, N10 -methylene tetrahydrofolate reductase has been achieved by ammonium sulfate fractionation, protamine sulfate fraction, and by various column chromatography procedures. The two enzymes studied catalyze sequentially the reductive transfer of the hydroxymethyl group of serine to the N5 position of tetrahydrofolate forming N5 -methyl tetrahydrofolate. The products of these reactions were assayed by the incorporation of C14 from a one-carbon precursor i.e., L-beta-(C14)-serine, or N5 -methyl …

The Role Of Thiamine Pyrophosphate In Regulating The Level Of Gamma-Aminobutyric Acid In The Central Nervous System, Bryant L. Adams

Theses and Dissertations

The level of gamma-aminobutyric acid in the brains of white rats suffering from thiamine deficiency symptoms resulting from pyrithiamine or oxythiamine antagonism of thiamine or from thiamine deprivation was measured. The level of this amino acid was 17.5% lower in the brains of pyrithiamine-treated rats and 20.0% lower in the brains of thiamine-deprived rats than in the control rats. Daily injections of ten times the required amounts of vitamin B6 in rats treated with pyrithiamine or oxythiamine or by thiamine deprivation had no effect on the level of brain gamma-aminobutyric acid or on the psiological behavior of these rats. The …

Studies Of Rat Brain Thiamine Pyrophosphokinase, Lavell Rolfson Johnson

Theses and Dissertations

The enzyme ATP:thiamine pyrophosphotransferase (thiamine pyrophosphokinaseJ was purified from acetone powders which were prepared from rat brains up to the chromatography on alumina C_γ step by a modified procedure that Mano (Y. Mano, J. Biochem., 42, 283 (1960)] developed for the purification of the rat liver thiamine pyrophosphokinase. The enzyme was purified 12 fold over that in the original extract of the acetone powder. A spectrophotometric assay for the brain thiamine pyrophosphokinase was developed utilizing brewer's yeast apotransketolase. The transketolase assay was able to detect less than 10^-11 moles of thiamine diphosphate, which was synthesized by the kinase, in the …

The Vitamin B1-Sparing Action Of Sorbitol, Gisela Eleanor Bethsold

Theses and Dissertations

The present study was initiated to determine the effect sorbitol has upon the enzymatic functions of thiamin; specifically, what effect sorbitol bas upon the alpha-ketoglutarate and pyruvate dehydrogenases activities of rat liver mitochondria. In addition, the effect of varying the dietary sorbitol concentration on the development of an athiaminosis was studied, particularly the effect on growth rate.

Studies On The Biological Oxidation Of Ferrous Iron And Chalcopyrite, Loral Alvin Decker

Theses and Dissertations

The purpose of this investigation was to determine the effectiveness of the leaching streams from waste copper ore dumps as a nutrient solution for subsequent biological oxidation and solubilization of sulfide minerals. The copper in the leaching streams is precipitated by displacement with iron. As a result of this procedure the solution is high in ferrous iron concentration and has a pH of near 3.7. The bacteria in the leaching streams are capable of oxidizing ferrous iron to the ferric state and sulfide minerals to sulfates. The effects of temperatures, pH, urea, and aeration rate on the oxidation of the …

A Study Of Three Photochemical Activities Of Chloroplasts, Jimmy W. Hinkson

Theses and Dissertations

Chloroplasts of sugar beet leaves were subjected to treatment by heating, by incubation at pH 8.5 and pH 5.5, by incubation in ion low water, by sonic oscillation, by digestion with enzymes (lysozyme, pancreatin, and lipase), by extraction with digitonin, and by chemical compounds including 8-hydroxy-quinoline, sodium azide, zinc sulfate, phenyl mercuric acetate, potassium cyanide, 1-allylthiourea, 2,4-dinitrophenol, thymol, and p-chloromercuribenzoate. A comparative study was made of the effects of these treatments upon three photochemical reactions (the photoreduction of 2,6- dichlorophenol indophenol, the photoreduction of indigo carmine in the presence of a couple composed of 2,6-dichlorophenol indophenol and ascorbic acid, and …

Chemosynthetic Microorganisms In Leaching Sulfide Minerals, A. Keith Jameson

Theses and Dissertations

This investigation showed that a single bacterium was capable of sulfide oxidation in exposed ore bodies. This single bacterium was isolated and characterized. It exhibited different characteristics than any previously known organism. It was found to be a chemosynthetic autotroph. An optimum nutrient concentration was determined for pyrite oxidation. The nutrient concentration was determined for pyrite oxidation. The nutrient contains only a source of phosphate and nitrogen. Urea was found to be a better source of nitrogen than ammonium sulfate which had previously been used. The effect of various organic compounds on the oxidation of pyrite was observed. Glucose and …

Oxidation Of Molybdenite With The Aid Of Microorganisms, Ralph Anderson

Theses and Dissertations

This work represents a study on the biological oxidation of molybdenite, MoS2. The principal objective of this study was to investigate the possibility of the biological oxidation of molybdenite, to determine the physical and chemical conditions under which the oxidative process occurs, and to develop a strain of bacteria or alter the activity of the microorganisms by acclimatization to increase their activity on sulfide minerals, in particular molybdenite. Minerals used in this study were pyrite (FeS2), chalcopyrite (CuFeS2), molybdenite concentrate, and molybdenite ore. The microorganisms used in this investigation were autotrophic bacteria obtained from the leaching streams of Bingham Canyon, …

Photo-Oxidations Sensitized By Physiologically Important Substances, Ernel D. Ihnen

Theses and Dissertations

A study of the photo-oxidation of the 2,6-dichlorophenol indophenol-ascorbic couple was undertaken with a view to investigating the postulated mechanism of oxidation by a component of the oxygen evolving sequence operative in photosynthesis. A spectrophotometric method for the measurement of the progress of photo-oxidation was developed. Temperature and pH dependence were determined and the effect of various common inhibitors and physiological substances tested. None of the common Hill reaction inhibitors were effective in the ascorbic acid photo-oxidation in the system used. KCN and catalase as well as a few other high molecular weight proteins were able to inhibit dark auto-oxidation …