Life Sciences Commons^™

Methemoglobin Reduction In The Presence Of Various Concentrations Of Oxyhemoglobin, Ali Mansouri

Journal of the Arkansas Academy of Science

Methemoglobin reduction in human red blood cell is governed by many factors, one of which is the concentration of oxyhemoglobin. Experiments were conducted to determine the inhibition of methemoglobin reduction by oxyhemoglobin under concentrations approaching those in the red cell. Hemoglobin, methemoglobin reductase and cytochrome b₅ were prepared by routine protein purification methods. Methemoglobin reduction was carried out at 37° C in spectrophotometer. It is shown that methemoglobin concentration variation (at least when it is above 1 mM) does not change the reduction rate. Whereas, the ratio of methemoglobin to oxyhemoglobin determines the rate of methemoglobin reduction when all other …

Demonstration Of A Heat-Stable Cyclic Gmp Phosphodiesterase In The Medium Of Physarum Flavicomum, Thomas J. Lynch, Mark S. Ross

Journal of the Arkansas Academy of Science

No abstract provided.

Role Of Centriolar Matrix And Striated Rootlets In Centriolar Pairing And Orientation During Spermatogenesis In Hydractina Echinata, Maurice G. Kleve

Journal of the Arkansas Academy of Science

Transmission electron microscopy of the spermatogenic stages of the hydroid, Hydractinia echinata, reveals a series of complex structural and positional changes in the centrioles of spermatocytes and spermatids. The newly generated centriolar pairs of spermatocytes form an unusual four-centriole aggregate that persists until cell division. The distal centrioles of this aggregate are shrouded with a very dense matrix that accumulates after centriolar replication. This matrix facilitates the mechanical attachment between distal centrioles and microtubular nucleating satellites, striated rootlets and pericentriolar processes. The association of these accessory structures occurs sequentially and is repeated in spermatocytes and spermatids. An electron dense plaque, …

Cloning And Characterization Of A Gene (Fadr) Involved In Regulation Of Fatty Acid Metabolism In Escherichia Coli, Concetta C. Dirusso, William D. Nunn

Department of Biochemistry: Faculty Publications

The regulatory gene fadR has been previously characterized by classical genetic means as a diffusible protein which exerts negative control over fatty acid degradation and acetate metabolism. fadR has also been implicated in the regulation of unsaturated fatty acid biosynthesis. To facilitate the identification of the product of the fadR gene and to study the mechanism by which this multifunctional regulatory gene exerts its control, we cloned a segment of DNA containing the fadR gene in the phage vector λL47. Subsequent subcloning of a segment of the chromosomal DNA from the λfadR⁺ phage into various plasmid vectors resulted in …

Proliferation Of Schwann Cells Induced By Axolemmal And Myelin Membranes, Michael Paul Dinneen

Theses and Dissertations

Purified Schwann cells were cultured from neonatal rat sciatic nerve using a modification of the method of Brockes (Brockes, J.P. et al, Brain Res. 165(1979) 105-118). Schwann cells and contaminating fibroblasts were unambiguously identified using fluorescent antibodies to 2'3' cyclic nucleotide 3'-phosphodiesterase and the thy 1.1 antigen respectively. The Schwann cells were quiescent unless challenged with mitogens, They proliferated rapidly in response to the soluable mitogen, cholera toxin, or to membrane fractions from rat CNS or PNS, prepared by the method of DeVries (DeVries, G.H., J Neurochem, 40 (1983) 1709-1717). Mitogenic activity was present in both axolemmal and myelin enriched …

Transition-State Analog Affinity Purification Of Proteases, Arunbhai Patel

Dissertations

No abstract provided.

Bionomics Of Culicoides Spp. (Diptera: Ceratopogonidae) From A Coastal Georgia Barrier Island, George J. Magnon

College of Graduate Studies: Theses & Dissertations (1964–2006)

The seasonal abundance of adult Culicoides spp. on a coastal Georgia barrier island was determined during a 20-month period, September 1983 through May 1985 by CO₂-baited CDC miniature light traps and sticky cylinder traps.

A total of 220,670 adult biting midges representing nineteen species and including the first report of C. loughnani Edwards in Georgia was collected. Culicoides furens (Poey), C. hollensis (Melander and Brues), and C. melleus (Coquillett) were the most abundant. Culicoides furens was abundant from early spring to late fall. Culicoides hollensis and C. melleus were bimodal with spring and fall abundance, but C. melleus …

Aspects Of Mitochondrial Oxidative Metabolism, Leslie Kushner

Dissertations, Theses, and Capstone Projects

The effects of metabolic intermediates and coenzymes on the activities of fatty acyl-CoA synthetase and carnitine palmitoyltransferase (CPT) in rat heart mitochondria were studied. ADP and palmitoyl-l-carnitine were weak inhibitors of fatty acyl-CoA synthetase. CPT A was inhibited 59.3% by 100 (mu)M succinyl-CoA. However, succinate thiokinase activity in rat heart cytosol was too low to maintain a succinyl-CoA concentration sufficient to affect the activity of CPT A. There was no indication that fatty acid oxidation in rat heart mitochondria is controlled via the regulation of either CPT A or fatty acyl-CoA synthetase.

The dependence of the oxidation of unsaturated fatty …

Characterization Of Kpni Interspersed, Repetitive Dna Sequence Families And Their Association With The Nuclear Matrix, Joseph A. Chimera

Electronic Theses and Dissertations

The KpnI, 1.2 and 1.5 kb families of interspersed repetitive DNAs from the African green monkey genome were isolated and characterized. Each family contains three populations of segments based on their sequence lengths and susceptibility to cleavage by the restriction enzymes KpnI and RsaI. The first population contains the smallest segments which are susceptible to both KpnI and RsaI cleavage and have fragment lengths of 1.2 kb (1.2 kb family) and 1.5 kb (1.5 kb family) respectively. The members in this population are referred to as KpnI-sensitive segments. The second population contains longer segments (> 2 kb) which represent fusions …

Correlation Studies Between Human Seminal Plasma Trace Metal Concentrations And Semen Quality, De-Shen Mei

Chemistry & Biochemistry Theses & Dissertations

Methodologies for the determination of cadmium, copper, selenium and zinc concentration in human seminal plasma have been developed using polarized Zeeman-effect atomic absorption spectroscopy. Within-run coefficients of variation (CY's) for cadmium, copper, selenium and zinc from pooled seminal plasma were found to be 8%, 5% , 4% and 0.5% at a mean (SD) concentration of 0.48 (0.04), 147 (7 .6), 53 (2. 1) and 122,900 (700) μg/ℓ, (n = 10), respectively.

Between-run CV's were determined to be 4% , 7%, 5% and 7% at a mean (SD) concentration of 2.4 (0.1), 147 (10. 1), 67.7 (3.4) and 115,800 (8300) …

Negative Cooperatively In The Binding Of 3-Aminopyridine Adenine Dinucleotide To L-Α-Glycero-3-Phosphate Dehydrogenase, David Wheeler

Chemistry & Biochemistry Theses & Dissertations

Glycerol-3-phosphate dehydrogenase catalyzes the reversible reaction:

NAD⁺ + G-3-P ⇌ NADH + DHAP + H⁺

The enzyme is found in organisms from E. coli to man. The primary function of the enzyme varies from organism to organism. Depending on the metabolism of the organism, the enzyme may function primarily to generate G-3-P for lipid (in organisms with a highly aerobic biosynthesis metabolism), or it may function primarily to produce NAD⁺ to support glycolysis. Complex multicellular organisms possess tissue specific isozymes of GPDH whose kinetic parameters reflect the relative importance of G-3-P and NAD⁺ production in various …

1-0-Alkyl-2-Acetyl-Sn-Glycero-3-Phosphorylcholine (Platelet Activating Factor): Interactions With Sheep Platelets, Duane O. Beaumont

Loma Linda University Electronic Theses, Dissertations & Projects

Platelet Activating Factor (PAF) seems to link platelet function with allergy and inflammation. Early events associated with the action of PAF on platelets are poorly understood, but PAF appears to interact with a platelet membrane receptor. Two photoaffinity-labelling analogs of PAF, diazoacetyl PAF and trifluorodiazoacetyl PAF, were designed for examining the interaction between PAF and its platelet receptor. These photoactive compounds were selected on the basis of the structure-activity relationship of PAF, and the general guidelines of the photoaffinity labelling technique. Some progress has been made in their synthesis and characterization.

The standard platelet aggregation assay was unreliable in testing …

Extrathyroidal Human Biochemistry Of Monoiodotyrosine, Stanley Andrew Tan

Loma Linda University Electronic Theses, Dissertations & Projects

The existence of extrathyroidal monoiodotyrosine (MIT) has been debated because of uncertain results from measuring serum MIT. The reason is that the isotopic and chromatographic assays used have not been sensitive enough. This investigation undertook to establish the existence of MIT in serum utilizing a radioimmunoassay (RIA), to study the physiology of serum MIT, and to discover the pharmacologic effects of exogenous MIT.

MIT-binding antibody was raised in rabbits by immunization with an MIT-porcine thyroglobulin conjugate. The antiserum bound 40% of the labeled MIT at a final dilution of 1:1000. Incubation of 100 µ1 antiserum with 50 pg [^{125 …}

The Effects Of Tri-O-Cresyl Phosphate On 2',3'-Cyclicnucleotide-3'-Phosphohydrolase Activity In Neural And Non-Neural Tissue Of The Rat, Mark K. Schlager

Chemistry & Biochemistry Theses & Dissertations

A total of 44 Long-Evans rats received 5.7 g/kg tri-O-cresyl phosphate (TOCP) orally every 3 days for 6 days. A second group of 13 rats received 4.6 g/kg orally every 3 days for 6 days. Animals were administered atropine sulfate (10 mg/kg) intraperitoneally every 6 hours or as needed to alleviate cholinergic symptoms. Ten control animals were given corn oil orally. Eight rats (18%) survived the 5.7 g/kg dose regimen and 5 rats (38%) survived the 4.6 g/kg dose regimen. Surviving rats were killed at 30-33 days post-dose. Signs of delayed neuropathy included ataxia and motor impairment.

The CNP activity …

Two-Dimensional Electrophoretic Studies Of Normal Spematozoal Proteins, Ying Lin

Chemistry & Biochemistry Theses & Dissertations

Protein profiles of human spermatozoa have been mapped by using high resolution two-dimensional electrophoresis which combines isoelectric focusing as the first dimension and polyacrylamide gradient gels electrophoresis in SDS as the second dimension. The profiles of normal spermatozoa! extraction, acrosomal extraction and seminal plasma showed high resolution in its separation capability. No qualitative or quantitative differences were observed by using Coomassie Brilliant Blue R-250 staining in a much higher concentration of proteins. However, coomassie blue lacks the sensitivity to detect many minor components. Therefore, a silver staining procedure was used, which resulted in greatly increased sensitivity.

Samples from normal fertile …

Photoaffinity Labeling Of The Antimycin Binding Site In Rhodopseudomonas Sphaeroides, Emily Wilson

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The purpose of this study was to identify the site of interaction of antimycin with the ubiquinone-cytochrome b-c1 oxidoreductase in the photosynthetic bacteria, Rhodopseudomonas sphaeroides. To accomplish this goal, three areas of research were undertaken: the synthesis of a radiolabeled, photoaffinity analog of antimycin, identification of the inhibitory characteristics of this analog, and the photoaffinity labeling of the antimycin binding site. All three areas were accomplished.

The major finding of this study was the identification of an 11,000 dalton polypeptide as the predominantly labeled protein. Although this polypeptide was not exclusively labeled, it was consistently labeled and showed competition …

Time Course Of Enzyme Catalyzed Reactions: The Stoichiometry A --> P + Q, Tamra Tuttle

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The feasibility of using an integrated rate equation to analyze the kinetics of a second-order, enzyme-catalyzed reaction has been investigated. The inducible arginine decarboxylase from Escherichia coli B. was chosen for this study because it catalyzes an irreversible reaction with stoichiometry A --> P + Q, the simplest second order case. Values for five of the eight possible kinetic constants were determined from 21 time courses. Of the remaining three, the uncompetitive product inhibition constant for CO₂ was shown to be between 0 and 0.06, while the values of the competitive product inhibition for CO₂ and the uncompetitive …

Investigation Of The Coenzyme Binding Site Of Lactate Dehydrogenase From Chicken Muscle, Jia-Hwei Lin

Chemistry & Biochemistry Theses & Dissertations

Investigation of the coenzyme binding site of lactate dehydrogenase was approached by using coenzyme analogs. Inhibition studies of this enzyme using n-alkylphosphates as coenzyme analogs were performed with a fluorometric method. The results showed that all of the n-alkylphosphates were competitive inhibitors with respect to NAD⁺.

The n-alkyl-ADPs were synthesized from AMP and dicycloheaylammonium salts of n-alkylphosphates. The synthetic compounds were subjected to NMR spectroscopic studies, thin layer chromatography and determination of the ratio of adenosine moiety to phosphate groups. The results indicated that the synthetic compounds were n-alkyl-ADPs.

These synthetic compounds were used as coenzyme analogs in …

A Method For The Comparison Of Hdl Containing Apoprotein E And Hdl Cholesterol, Steven A. Knizner

Chemistry & Biochemistry Theses & Dissertations

Human serum high density lipoproteins (HDL) were isolated by preparative salt density gradient ultracentrifugation and analyzed further by heparin-sepharose affinity chromatography. Separation of the major classes of serum lipoproteins was achieved after a single ultracentrifugation on a salt density gradient at 272,000 x g for 36 hours in a swinging bucket rotor. After isolation, the total HDL was subdivided into HDL containing apoprotein E and HDL without this apoprotein by heparin-sepharose affinity chromatography. Relative amounts of these subclasses were then calculated.

HDL-cholesterol was measured after precipitation of LDL and VLDL by phosphotungstic acid and magnesium. The statistical relationship between HDL …

Purification And Characterization Of A Camp- And Ca2+-Calmodulin-Independent Glycogen Synthase Kinase From Porcine Renal Cortex, Stephen J. Beebe, Erwin M. Reimann, Keith K. Schlender

Bioelectrics Publications

We recently reported the partial purification of a cAMP-independent and Ca²⁺-calmodulin-independent glycogen synthase kinase from porcine renal cortex (Schlender, K. K., Beebe, S. J., and Reimann, E. M. (1981) Cold Spring Harbor Conf. Cell Proliferation, 389-400). Subsequent purification indicated that the enzyme preparation consisted of at least three forms of glycogen synthase kinase which could be resolved by ATP gradient elution from aminoethylphosphate-agarose (AEP-agarose). The predominant form of glycogen synthase kinase, which eluted from AEP-agarose between 2 and 6 mM ATP, was purified approximately 800-fold and is designated GSK-A1. It had a molecular weight of 45,000-50,000 as determined …

Some Plant Leaves Have Orientation-Dependent Epr And Nmr Spectra, Douglas C. Mccain, Ted C. Selig, Govindjee, John L. Markley

Faculty Publications

Proton nuclear magnetic resonance (¹H NMR) spectra of leaves from 50 plant species were obtained at a spectrometer frequency of 470 MHz. Water present in leaf samples gives rise to characteristic spectral patterns. Most species show only one broad ¹H NMR peak; however, the leaves of some plants display complex, orientation-dependent spectra in which a common three-line pattern is discerned. The pattern varies with the angle between the leaf surface and the external magnetic field. Proton relaxation measurements show the presence of at least two water compartments in the leaves. The compartments are responsible for different components …

Potassium 1,3,3a,8a-Tetrahydro-3a,8a-Dihydroxy-2,8-Dioxoindeno[1,2-D]Imidazole-2-Ethanesulfonate, K+.C12h11n2o7s- / By D. D. Bray And D. D. Clarke Chemistry Department, Fordham University, Bronx, Nyew York 10458, Usa And N. Chatterjie And B. Sinha Institute For Basic Research In Development Disabilities, 1050 Forest Hill Road, Staten Island, New York 10314, Usa, Diana Bray, Donald Dudley Clarke Phd, Nithiananda Chatterjie, B. Sinha

Chemistry Faculty Publications

Mr = 366·40, monoclinic, P2/c, Z = 4, a=6·195 (1), b= 17·701 (4), c= 1.2·864 (3)A, P= 116·25(2)0 , V=l387·8A3, Dm=1·74, Dx= 1·754 Mg m-3, CuKa, l = 1·5418 A, ,u = 5·07 mm-1, T= 298 K, F(OOO) = 752, R = 0·074 for 2091 observed independent intensity data. Crystals were prepared by reacting the K salt of ureidotaurine with ninhydrin in acid solution. The product shows that the unsubstituted N -of the ureidotaurine reacts with the central C of ninhydrin

Cerebral Glutamine/Glutamate Interrelationships And Metabolic Compartmentation / S. Berl And D. D. Clarke, Soll Berl, Donald Dudley Clarke Phd

Chemistry Faculty Publications

The fact that in animals the concentration of glutamate and glutamine are higher in brain than in any other tissue initially attracted interest in attempting to understand the function or functions of these amino acids in the central nervous system. They were evidently required for protein synthesis and early studies established their close relationship to the Krebs tricarboxylic acid cycle and energy metabolism in brain. Glutamine formation in brain was also shown to be sensitive to elevations in plasma ammonia levels and to serve as the primary method for detoxifying this substance in brain in contrast to urea formation in …

The Role Of Recognition In Plant Disease, J. M. Daly

Department of Biochemistry: Faculty Publications

As noted by Sequeira (110,111), the term recognition has become very popular in the literature of host-pathogen interactions over the last ten years. Despite recent facile usage, its meaning is not always clear to me. Recognition is defined by Clarke & Knox (16) as the "initial event in cell-cell communication that elicits a defined biochemical, physiological, or morphological response." This requires the not-so-easy task of defining "initial event" and "communica­ tion" with precise chemical or physical mechanisms. For pathologists, Se­ queira (110) has provided a more restricted meaning: "An early specific event that triggers a rapid, overt response by the …

Microheterogeneity Of Type Ii Camp-Dependent Protein Kinase In Various Mammalian Species And Tissues, Alison M. Robinson-Steiner, Stephen J. Beebe, Stephen R. Rannels, Jackie D. Corbin

Bioelectrics Publications

Excluding autophosphorylated species, at least six forms of the regulatory subunit of type II cAMP-dependent protein kinase (R(II)) from various mammalian tissues were identified by sodium dodecyl sulfate (SDS) gel electrophoresis of purified samples and of crude preparations photoaffinity labeled with 8-azido[32P] cAMP and by gel filtration. After autophosphorylation some heart R(II) forms termed type IIA (bovine, porcine, equine, and dog) shifted to a more slowly migrating band on SDS gels while others termed type IIB (rat, guinea pig, rabbit, and monkey) did not detectably shift. Both subclasses of R(II) exhibited variation in apparent M(r) on SDS gels. Bovine and …

Two Classes Of Camp Analogs Which Are Selective For The Two Different Camp-Binding Sites Of Type Ii Protein Kinase Demonstrate Synergism When Added Together To Intact Adipocytes, Stephen J. Beebe, Rob Holloway, Stephen R. Rannels, Jackie D. Corbin

Bioelectrics Publications

Twenty-five cyclic nucleotide analogs were tested individually to act as lipolytic agents and to activate adipocyte protein kinase. The lipolytic potency of individual analogs correlated better with their K(a) for protein kinase and their lipophilicity rather than with either parameters alone. Some of the most potent lipolytic analogs had high I50 values for the particulate low K(m) cAMP phosphodiesterase suggesting that their effect was not due to raising endogenous cAMP levels through inhibition of phosphodiesterase. The most potent lipolytic analogs contained a thio moiety at the C-8 or C-6 position. These analogs exhibited concave upward dose-response curves. At high concentrations …

Comparison Of Surface Polysaccharides Of R. Leguminosarum Anu57 (Exo-1), R. Leg. Anu57(Pbr1an) (Nod+, Nif-), And R. Leg. Anu57(Pjb5j1) (Nod+, Nif-), Frank G. Hustmyer

Masters Theses

Extracellular polysaccharides (EPS) and lipopolysaccharides (LPS) are suspected to be involved in the legume/Rhizobium symbiotic process. Polysaccharide compositions from the EPS, LPS, capsular polysaccharide (CPS), and small polysaccharide (SmPS) were compared between Rhizobium leguminosarum ANU57 (parent, Exo^-1) and two mutants R. leg. ANU57(pBR1AN) (nod⁺, nif^-) and R. leg. ANU57(pJB5J1) (nod⁺, nif^-) both containing a Sym (symbiotic) plasmid insertion. R. leg. ANU57(pBR1AN) and R. leg. ANU57(pJB5J1) are nod⁺ on clover and peas, respectively. The nod⁺ mutants produce larger amounts of EPS and decreased amounts of LPS compared to …

Las Vegas Wash Advanced Water Quality Study: Final Report, Richard A. Roline, James J. Sartoris, U.S. Bureau Of Reclamation

Publications (WR)

The purpose of the Las Vegas Wash Advanced Water Quality Study is to determine the existence, extent, and mechanisms of nutrient and toxin stripping in Las Vegas Wash under present conditions and under future conditions, both with and without construction of the proposed salinity control unit as described by the Bureau of Reclamation (USBR, 1982b). This study was performed for the Lower Colorado Region Division of Planning by personnel of the Environmental Sciences Section of the Division of Research and Laboratory Services, E&R Center, Denver, Colorado. Work on the study began in February 1983. The general approach adopted for this …

Extracellular Phosphodiesterase From The Growth Medium Of The Myxomycete Physarum Flavicomum, Thomas J. Lynch, Mary E. Farrell

Journal of the Arkansas Academy of Science

The plasmodium of the myxomycete Physarum flavicomum secretes cyclic AMP phosphodiesterase into the medium. The extracellular enzyme had a pH optimum between 7 and 8 and a Km of about 500 μM cyclic AMP and was inhibited by theophylline, caffeine and 3-isobutyl-l-methyxanthine (MIX). A marked decrease of enzyme activity was noted in the presence of EDTA, suggesting the requirement of Mg+ + by the enzyme. Addition of Mg+ + and Ca + + stimulated the enzyme while Zn+ + , Co+ + , Pb+ + , Mn+ + , Fe + + + , Ni+ + , and Cu + …

Evaluation Of Peptide Assimilation In Ehrlich Ascites Tumor Cells, John M. Castellano

Biological Sciences Theses & Dissertations

The dipeptides glycyl-L-(³H)-leucine and L-phenylalanyl-L-(³H)-leucine were synthesized in radioactive form and utilized in the analysis of peptide transport and hydrolysis by Ehrlich ascites tumor cells. The tumor cells were cultured in the abdominal cavity of CF-1 albino mice and harvested into sterile isotonic buffer for all experimental analyses.

The results of this study reveal that the tumor cells rapidly accumulate radiolabel when incubated with the above mentioned peptides, but that the peptide is most probably hydrolyzed extracellularly and free L-(³)-leucine transported. This hypothesis is based on the observations of reduction in accumulation of radiolabel …