Life Sciences Commons^™

Optimal Conditions For Kinetic Study Of Succinate Dehydrogenase In Rat Liver, Collie B. Shaw, Tara L. Chronister, John D. Peck

Journal of the Arkansas Academy of Science

Succinate dehydrogenase (SDH) commonly is assayed as a marker enzyme for mitochondrial activity. The literature presents numerous conditions for conducting this assay due to the fact that, it has been difficult to get sufficient reduction of the acceptor dye, 2,3,5-triphenyl-2H-tetrazolium chloride (TTC). This study was undertaken to optimize the SDH-catalyzed reduction of TTC dye by evaluation of a greater range of molor ratios of TTC to succinate and by further evaluation of additives reported as beneficial. Improvement in enzyme specific activity was achieved by liver perfusion via the left cardiac ventricle with homogenizing solution. Increase in TTC from 1 to …

Investigation Of Nickle As A Matrix Modifier In The Atomic Absorption Analysis Of Tin, Charles William Saunders

Chemistry & Biochemistry Theses & Dissertations

Nickel has been widely used as a matrix modifier in the flameless atomic absorption analysis of arsenic and selenium. Its utility as a modifier for tin has been investigated using Polarized Zeeman effect Graphite Furnace Atomic Absorption Spectroscopy. Peak area, peak shape, and appearance times were compared in modified and nonmodified standards using the system computer's internal integration and a detector/readout system with 18 msec resolution.

The addition of nickel at 600 mg/L increased the tin absorbance signal five-fold. The improved signal to noise ratio resulted in a two-fold increase in the sensitivity and a lower limit of detection. The …

Formation Of Influenza Virus Particles Lacking Hemagglutinin On The Viral Envelope, Asit K. Pattnaik, Donald J. Brown, Debi P. Nayak

School of Veterinary and Biomedical Sciences: Faculty Publications

We investigated the intraceUular block in the transport of hemagglutinin (HA) and the role of HA in virus particle formation by using temperature-sensitive (Is) mutants (1s134 and 1s61S) of inOuenza virus AlWSN/33. We found that at the nonpermissive temperature (39.5°C), the exit of ts HA from the rough endoplasmic reticulum to the Golgi complex was blocked and that no additional block was apparent in either the exit from the Golgi complex or post-Golgi complex transport. When MDBK ceUs were infected with these mutant viruses, they produced noninfectious virus particles at 39.5°C. The efficiency of particle formation at 39.5°C was essentiaUy …

Quantitation And Identification Of Organic N-Chloramines Formed In Stomach Fluid On Ingestion Of Aqueous Hypochlorite, Frank E. Scully Jr., Katherine Mazina, Daniel E. Sonenshine, Frederick Kopfler

Biological Sciences Faculty Publications

The chemical reactions that hypochlorite undergoes in the body when chlorinated water is ingested have received very little attention. Because amino nitrogen compounds are important components of the average diet, the reactions of hypochlorite with amino compounds in the stomach were investigated. Stomach fluid was recovered from Sprague-Dawley rats that had been fasted for 48 hr and administered 4 mL deionized water. The chlorine demand of the stomach fluid was determined. An average volume-independent demand of 2.7 mg chlorine was measured. At doses below 40 mg/L chlorine reducing reactions appeared to account for reduction of all oxidizing species within 15 …

Solid Phase Extraction Of Mammalian Cell Mitochondrial Dna And Its Electrophoretic Separation In Agarose Gels, Theresa Brick-Miller

Biological Sciences Theses & Dissertations

An improved and efficient electrophoretic procedure for the isolation of mitochondrial DNA (mtDNA) is described. The solid phase extraction procedure for the isolation of mtDNA involves embedding as few as 200,000 Ehrlich ascites tumor cells into a block of agarose, digestion of cellular membranes by detergent action and subsequent electrophoretic separation of nucleic acids in agarose gels. Genomic DNA remains in the original block of agarose while the mtDNA migrates in the separation gel to a position equivalent to that obtained for mtDNA isolated by traditional procedures.

The efficacy of a large number of detergents, agarose concentrations, embedding conditions, embedding …

Research Note: Getting Graphic: Computer Graphics In Biochemistry, Wilmon B. Chipman

Bridgewater Review

No abstract provided.

Isolation And Characterization Of Temperature-Sensitive Protein Synthesis Mutants Of Escherichia Coli By Directed Mutagenesis Of The Defective Bacteriophage Lambda Fus2, Kenton L. Lohman

Electronic Theses and Dissertations

Mutagenesis of the defective transducing bacteriophage lambda fus2 was used to isolate a collection of temperature-sensitive mutants of E. coli in the major ribosomal protein gene cluster. Four mutants were examined in detail. Two of the mutants were resistant to the ribosomal antibiotics neamine and spectinomycin. Another mutant was defective in 50S ribosomal subunit assembly at 42(DEGREES)C. The 30S subunit proteins S17 and S19 were changed in two different mutants. Each protein migrated as a more basic species in two-dimensional gels of ribosomal proteins. Ribosomes from each of the four mutants examined showed a temperature-dependent reduction in translational activity in …

Modification Of Chromium(Vi)-Induced Dna Damage By Glutathione And Cytochromes P-450 In Chicken Embryo Hepatocytes., Doreen Y. Cupo, Karen E. Wetterhahn

Dartmouth Scholarship

The role of glutathione and cytochrome P-450 in the production of DNA damage by chromium(VI) was examined in chicken embryo hepatocytes by the alkaline elution technique. Cellular levels of glutathione and cytochrome P-450 were altered by treating the hepatocytes with N-acetyl-L-cysteine, buthionine sulfoximine, isopentanol, or beta-naphthoflavone. A dramatic increase in chromium(VI)-induced DNA strand breaks was observed after increasing glutathione levels in the cells. Chromium(VI)-induced DNA strand breaks were even more numerous when the level of cytochrome P-450 was also increased. Upon depletion of glutathione levels and induction of cytochrome P-450 or cytochrome P-448, little or no DNA strand breaks or …

Determination Of Total And Unfilterable Plasma Aluminum In Uremic Patients, Theodore R. Delbridge

Chemistry & Biochemistry Theses & Dissertations

Aluminum accumulation in the soft tissues and bone of end stage renal failure patients has been described in the literature. In addition to aluminum normally encountered through ubiquitous sources, these patients are subject to exposures through the dialysate used for hemodialysis and through aluminum-containing phosphate binders. These increased exposures, associated with a decreased ability to excrete aluminum through the kidneys, lead to aluminum accumulation which may result in the onset of an encephalopathy syndrome and/or osteomalacic osteodystrophy. These complications may be very devastating.

Blood serum or plasma aluminum levels do not reflect the total body burden of aluminum. However, they …

Choline Inhibition Of Prothrombinase, Richard D. Leach

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of this study was to characterize the inhibitory effects of choline on the terminal enzymatic complex of the blood coagulation cascade. Reports in the literature have indicated that choline lengthens whole plasma clotting time with a likely site of inhibition being the step that converts prothrombin (II) to thrombin (IIa).

In order to quantitate the effects of choline on prothrombinase activity, bovine factor V and human factor II were isolated in this laboratory by established methods, and other prothrombinase components were obtained from commercial sources. The activity of venom-activated bovine factor Xa was assayed in the presence of …

Isolation And Characterization Of A Novel Motility Mutant In Salmonella Typhimurium, David Matthew Chacko

Loma Linda University Electronic Theses, Dissertations & Projects

A new mutant, BT40, isolated from mutagenized cultures of S. typhimurium was deficient in chemotaxis (Che^- phenotype) in tryptone semisoft agar, but had random motility with a tumbling frequency (37%) similar to that of the parental strain, ST23 (39%). In spatial assays accumulation of BT40 cells in capillary tubes containing attractants was decreased by 33%-59% compared to ST23, but BT40 had a normal response to attractants in temporal assays. The methylation level of the chemotactic proteins was decreased in BT40. At room temperature, the mean swimming speed was 11 μm/s for BT40 and 20 μm/s for ST23; more BT40 …

The Relationship Of Vitamins B-6 And B-12 To Plasma Homocysteine Levels In Men At Low- And High-Risk For Coronary Heart Disease, Marlene E. Swift

Loma Linda University Electronic Theses, Dissertations & Projects

There is evidence that homocysteine may be a factor in increasing the risk of coronary heart disease (CHD). To explore this relationship further, we assessed the interrelation of dietary vitamins B-6, B-12 and folate, and plasma pyridoxal-5'-phosphate and vitamin B-12 with plasma free and protein-bound homocysteine levels. Fasting blood and three-day dietary records were obtained from nine men low-risk (LR) and five at high-risk (HR) for CHD. The HR mean systolic blood pressure, total cholesterol (TC), and TC/cholesterol ratios were significantly higher than LR levels. Groups were comparable by age, weight, height, skinfolds, exercise, and smoking history. No significant difference …

X-Ray Synchrotron White Beam Excitation Of Auger Electrons, S. M. Durbin, L. E. Berman, B. W. Batterman, J. M. Blakely

Scanning Electron Microscopy

Auger electron spectra have been measured at the Cornell High Energy Synchrotron Source (CHESS), using the full white beam x-ray spectrum as the excitation source. Ordinary Auger spectra obtained in the laboratory with an electron beam source must employ derivative techniques to distinguish the Auger structures from the large background due to the excitation beam. The synchrotron white beam eliminates this source of background and produces signal rates as high as 10⁷ cps. Superior signal-to-background ratios are found for Auger peaks above a few hundred eV, and count rates are large enough to suggest microprobe applications. X-ray induced Auger …

Research Note: Muscle Biochemistry From Oxford University, Vahe Marganian

Bridgewater Review

No abstract provided.

A Biochemical And Genetic Analysis Of The American Shad Alosa Sapidissima Migrating Into The Chesapeake Bay, Edward Norbert Sismour

Biological Sciences Theses & Dissertations

Seven enzymes extracted from heart and liver tissues of American shad Alosa sapidissima captured during migrations into the Chesapeake Bay and Nansemond River were electrophoretically surveyed. Malate dehydrogenase and nonspecific esterase displayed polymorphic loci. The esterase polymorphism was used as a biochemical genetic marker to analyze the shad migration through the southern Chesapeake Bay. The genetic composition of the shad migration changed significantly over time indicating that this migration is probably composed of multiple, genetically divergent spawning populations. Results of this study are subject to differing interpretations. Observed temporal heterogeneity may result from separation of northern and James River stocks …

Acetylcholinesterase In The Neonatal Rat Brain: The Effects Of Prenatal Ethanol Exposure, Linda M. Boland

Biological Sciences Theses & Dissertations

Acetylcholinesterase (AChE) activity in the neonatal rat brain was determined by histochemical localization and quantitative biochemical analysis. AChE-positive perikarya were concentrated in the neostriatum, globus pallidus, medial habenular nucleus, amygdaloid complex, various hypothalamic nuclei, gigantocellular and paragigantocellular reticular nuclei, raphe nuclei, locus coeruleus, and motor nuclei of cranial nerves III-VII and IX-XII. Some areas stained differently or not at all when compared with the adult rat brain. The effects of prenatal ethanol exposure on regional AChE activity were assessed by comparing enzyme levels in offspring exposed via maternal consumption of a liquid diet containing 35 percent ethanol derived calories. This …

Human Lung Tryptase Purification And Characterization, Timothy J. Smith

Electronic Theses and Dissertations

Human lung tryptase (HLT), a mast cell derived trypsin-like enzyme, was isolated from whole human lung tissue obtained at autopsy. Increased yields from this purification process allowed extensive characterization of the enzyme. One of the critical steps in the purification scheme was the use of a linear heparin gradient to elute active material from cellulose phosphate. Gel filtration studies in 1.0 M NaCl yielded an apparent M(,r) of 135,000, and subsequent electrophoresis on sodium dodecyl sulfate-polyacrylamide gels demonstrated the presence of two active species with apparent M(,r) = 30,900 and 31,600. Enzymatic activity was sensitive to NaCl concentrations above 0.05 …

Autoregulation Of Collagenase Production By A Protein Synthesized And Secreted By Synovial Fibroblasts: Cellular Mechanism For Control Of Collagen Degradation, Constance Brinckerhoff, Mary Benoit, William Culp

Dartmouth Scholarship

No abstract provided.

Lectins, El-Nasir Lalani, M Wells

Department of Pathology and Laboratory Medicine

No abstract provided.

Acetylation-Deacetylation Of Synaptosomal Proteins: Effect Of Na+ / S. Berl, A. Colon And D. D. Clarke. Mt. Sinai School Of Medicine, New York, N.Y. 10029, Soll Berl, Arlene D. Colon, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Synaptosomes (S) when incubated with 3H-acetate (HA), are rapidly labeled. The perchloric acid (PCA) precipitable proteins contain most of the radioactivity probably as the acetyl moiety, which is released in a volatile form after acid or base hydrolysis (Berl et al., - J. Neurochem. 40:176, 1983). Much of the radioactivity was chased by addition of unlabeled acetate suggesting that the acetylation was reversible. Labeling was also decreased in the presence of veratridine/veratrine (V, 100uPt!) and this effect was prevented by tetrodotoxin (2uM). When Na was omitted from the incubation medium, the incorporation of (HA) was increased; however, the effect of …

Concanavalin A-Binding Enzymes Of Crotalus Scutulatus Scutulatus Venom, Michael W. Hinson, C. K. Childs, Bob D. Johnson, Dewey H. Sifford

Journal of the Arkansas Academy of Science

Crotalus scutulatus scutulatus crude venom was separated into two fractions by Concanavalin A Sepharose 4B affinity chromatography. The proteins binding to Con A exhibited phosphomonoesterase (orthophosphoric monoester phosphohydrolase EC 3.1.3.2), phosphodiesterase, 5'-nucleotidase (5'-ribonucleotide phosphohydrolase EC 3.1.3.5), phospholipase A(phosphatidate 2-acylhydrolase EC 3.1.1 .4), hyaluronidase (hyaluronate glycanohydrolase EC 3.2.1 d), N-benzoyl-L-arginine ethyl esterase, p-toluenesulfonyl-L-arginine methyl esterase, L-amino acid oxidase (L-amino acid: 02 oxidoreductase [deaminating] EC 1.4.3.2), and caseinolytic activities. Thrombin-like and NAD nucleosidase (5'-ribonucleotide phosphohydrolase EC 3.1.3.5) activities were not observed. The crude venom and the fraction containing the glycoproteins which bound to Con A were fractionated by DEAE Sephadex A-50 ion …

Nonsense Mutations In The Chlamydomonas Chloroplast Gene That Codes For The Large Subunit Of Ribulosebisphosphate Carboxylase/Oxygenase, Robert J. Spreitzer, Michel Goldschmidt-Clermont, Michele Rahire, Jean-David Rochiax

Department of Biochemistry: Faculty Publications

The Chlamydomonas reinhardtii chloroplast mutants 18-SB and 18-7G lack both the chloroplast-encoded large subunit and nuclear-encoded small subunit of the chloroplast enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39). A chloroplast intergenic-suppression model has been postulated to account for the genetic instability of 18-5B revertants. Here, we have determined the molecular basis of the 18-SB and 18-7G mutants. They contain nonsense mutations close to the 3' and 5' ends of their large-subunit genes, respectively. Puls-chase experiments revealed that the 18-SB mutant produces a truncated large subunit that is unstable. In connection with previous experiments, this work identifies nonsense suppression in the chloroplast. Small …

Camp-Dependent Protein Kinase Activation Lowers Hepatocyte Camp, Jackie D. Corbin, Stephen J. Beebe, Peter F. Blackmore

Bioelectrics Publications

Rat hepatocyte protein kinase was activated by incubating the cells with various cAMP analogs. Boiled extracts were then prepared and Sephadex G-25 chromatography was carried out. The G-25 procedure separated the analogs from cAMP since the resin had the unexpected property of binding cyclic nucleotides with differing affinities. Separation was necessary because the analogs would otherwise interfere with the sensitive protein kinase activation method developed for assay of cAMP. The cAMP analogs, but not 5'-AMP, lowered basal cAMP by 50-70%. The effect was rapid, analog concentration-dependent, and occurred parallel with phosphorylase activation, suggesting that the cAMP analogs act through cAMP-dependent …

Discriminative Insulin Antagonism Of Stimulatory Effects Of Various Camp Analogs On Adipocyte Lipolysis And Hepatocyte Glycogenolysis, Stephen J. Beebe, J. Bruce Redmon, Peter F. Blackmore, Jackie D. Corbin

Bioelectrics Publications

Although insulin effectively blocked hormone-stimulated glycerol output in adipocytes or phosphorylase activation in hepatocytes, the inhibitory effect of insulin on cAMP analog-stimulated cells depended on the cAMP analog used. Of the 20 analogs tested in adipocytes and 13 tested in hepatocytes, the effects of about half of them were effectively blocked by insulin, whereas the effects of many of them were not inhibited at all. In order to approach the explanation for this discriminative insulin action, the inhibitory effects of insulin on the responses to the analogs in the intact cells were correlated with the in vitro cAMP analog specificity …

Response Of A Nitrifying Biofilm To Copper, Denise Moldroski Mott

Masters Theses

In pure cultures of Nitrosomonas, copper is known to cause a stimulation of nitrification at low levels (≤ 0.03 mg Cu⁺²/1) and an inhibition affect at higher levels (≥ 0.05 mg Cu⁺²/1). Nitrifying activated sludges are much more tolerant of copper showing only 75% inhibition at 150mg Cu⁺²/1. The purpose of this paper was to determine the copper sensitivity of a nitrifying biofilm grown in a continuous flow trickling filter system.

A continuous flow trickling filter system was built using limestone chips for the biofilm substrate. The biofilm was established from secondary effluent. …

A Comparison Of The Acidic Extracellular Polysaccharides From Rhizobium Trifolii Mutants Affected In Trifolium Repens Root Hair Curling, Brian Andrew Hanley

Masters Theses

The compositions and sugar linkages of the acidic extracellular polysaccharide (EPS) from several Rhizobium trifolii mutants were compared. These mutants were derived from a wild type Rhizobium trifolii, ANU843, and are unable to cause normal root hair curling, Hac-, or nodulate clover, Nod-. Three of the mutants contain a Tn5 insertion in a 5.3 kb BglII fragment of the symbiotic plasmid (pSym). A fourth mutant is missing the pSym, which includes much of the 5.3 kb BglII fragment, both of these deletions result in Hac-Nod- phenotypes. When the 5.3 kb BglII fragment is cloned (pRT150) and reinserted into the …

The Biological Role Of Sialosyl Transferase Activity In Rat Brain, Robert Paul Durrie

Dissertations

No abstract provided.

Effects Of Growth Temperatures On The Fatty Acid Composition Of Isolated Chloroplasts From Two Species Differing In Heat Sensitivity, Roy Z. Gehring, Glenn W. Todd

Journal of the Arkansas Academy of Science

Lipid analyses of chloroplasts isolated from wheat (Triticum aestivum L. cv. Arthur) and milo (Sorghum bicolor cv. Funk's hybrid 522) suggest no major heat effect on lipid class distribution. Assuming milo is more heat tolerant than wheat and that increased saturated/unsaturated fatty acid values increase thermal stability, changes in sulfoquinovosyldiglyceride (SL) appear to be more important than phosphatidylglycerol (PG) in conferring thermal stability to isolated chloroplasts.

Effects Of Gamma Radiation On Salix Nigra Marsh Cuttings, Roy Z. Gehring

Journal of the Arkansas Academy of Science

Dormant cuttings of Salix nigra Marsh, were exposed to gamma radiation from a cobalt-60 source at a dose rate of 1110 roentgens (r) per minute. Exposure doses ranged from 50 r through 100 Kr with 100% mortality at 10 Kr. Low doses of ionizing radiation apparently stimulated growth, while higher doses progressively retarded growth. The growth of the staminate catkins was inversely proportional to total exposure dose (increased dose resulted in progressively decreased growth). Determination of growth rates at different intervals post-irradiation indicated recovery of the surviving fraction of the irradiated cuttings. At ninety days, the highest growth rate occurred …

Repair Of Lethal Damage Induced In A Hamster X Amphibian Hybrid Tissue Culture Cell Line By Ultraviolet Light And Gamma Ray, Susan Kulp, Ann Page, H. Gaston Griggs

Journal of the Arkansas Academy of Science

A hybrid cell line was formed by fusing cells from two established vertebrate tissue culture cell lines (V79B1 hamster and A84 Xenopus), which were known to differ substantially in resistance to some radiation-induced lesions and in associated repair potential. Ultraviolet and gamma ray dose-survival relations were then determined and analyzed for the three cell lines, primarily to compare the radiosensitivity (Do dose) of the hybrid line with that of each parental line for each radiation. Further experimentation was conducted to determine the extent to which observed differences in these Do doses could be attributed to interactions of the combined parental …