Biochemistry, Biophysics, and Structural Biology Commons^™

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 …

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 …

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 …}

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 …

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 …

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 …

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 Marine Biogeochemistry Of Selenium: A Re-Evaluation, Gregory A. Cutter, Kenneth W. Bruland

OES Faculty Publications

Vertical and horizontal profiles from the North and South Pacific Oceans demonstrate the existence of three species of dissolved selenium: selenite, selenate, and organic selenide (operationally defined). In surface waters, organic selenide makes up about 80% of the total dissolved selenium, selenite concentrations are uniformly low, and selenate concentrations rise with increased vertical mixing. The organic selenide maximum (thought to consist of seleno-amino acids in peptides) coincides with the maxima of primary productivity, pigments, bioluminescence, and dissolved free amino acids. Deep ocean waters are enriched in selenite and selenate, while organic selenide is nondetectable. In suboxic waters of the tropical …

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 …

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 …

Reduction Of Ferric Leghemoglobin In Soybean Root Nodules, Keuk-Ki Lee, Robert V. Klucas

Department of Biochemistry: Faculty Publications

Reduction of ferric leghemoglobin to ferrous leghemoglobin in soybean nodules (Glycine max [L.] Meff. cv Woodworth) was studied using a spectrophotometer equipped with an in-cell space diffuse reflectance accessory. Nodule slices prepared and scanned under nitrogen gas showed a ferrous leghemoglobin absorption spectrum. Nodule slices equilibrated with 100% O₂ or air exhibited two absorption bands characteristic of oxygenated leghemoglobin. The addition of CO shifted those bands to CO leghemoglobin absorption bands. Potassium ferricyanide was not effective in oxidizing ferrous to feffic leghemoglobin in nodule slices. However, ferric leghemoglobin was formed by treating the nodule slices with hydroxylamine, and …

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 …

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 …

High Resolution Two Dimensional Electrophoresis Of Soluble Proteins In Human Spermatozoa, Hsing-Ying Liu

Chemistry & Biochemistry Theses & Dissertations

The evaluation of human semen is very important for diagnosis of male infertility. Combined with the analysis of seminal plasma, a comprehensive biochemical screening of human spermatozoan proteins may be of assistance in providing qualitative and quantitative information to the physician to assist with evaluation.

In this study, human spermatozoa were ruptured by sonication. The soluble spermatozoan proteins were separated from the cellular debris by centrifugation and then analyzed in dissociated forms by two dimensional electrophoresis. The separated proteins were visualized by a silver stain method.

It has been shown that these combined techniques can resolve- four to five hundred …

Determination Of N,N'-Diphenylguanidine In Biological Tissues And Fluids Using High Pressure Liquid Chromatography, Edward Sidney Hunter Iii

Chemistry & Biochemistry Theses & Dissertations

N,N'-Diphenylguanidine (DPG) is a potentially toxic compound used in the vulcanization of rubber products. Distribution of DPG in hybrid mice (C57BL/6J interbred with DAB₂) was determined following intraperitoneal injection. Urine, fecal material, whole blood and tissues were collected at various times after administration of DPG and the extracts analyzed for DPG by high pressure liquid chromatography. Recovery of DPG from spiked urine and whole blood exceed 90%, however, the recovery of DPG from spiked tissue samples was approximately 70%.

DPG concentration in whole blood reached a maximum within ten minutes and rapidly decreased through 180 minutes after administration …

A Survey Of Glycosphingolipids In The Human Aorta: Their Relationship To Glycosphingolipids Of Blood And To Atherosclerosis, John L. Nappier

Dissertations

The four major glycosphingolipids, glucosyl ceramide, lactosyl ceramide, galactosyl lactosyl ceramide and globoside were isolated from both diseased and normal tissue from human aorta by solvent extraction followed by solvent partitioning and column chromatography. The levels of each were then determind quantitatively by high pressure liquid chromatography (HPLC) after derivatization. The individual glycosphingolipids were isolated by semi-preparative HPLC and the fatty acid composition of each was determined, after methanolysis, by gas chromatography.

In the diseased tissue, the amount of glycosphingolipid as well as the amount of crude lipid increased relative to the normal tissue. The relative amounts of cerebrosides increased …

Determination Of Erythrocyte Zinc Concentrations Using Zeeman Effect Atomic Absorption Spectroscopy, Tanya Maranda Lewis

Chemistry & Biochemistry Theses & Dissertations

The purpose of this research project was to develop assay procedures for determination of zinc in erythrocytes. Three methodologies were developed. The methods were evaluated on the basis of within-run and between-run precision. The most reproducible method, the 213.856 nm flame methodology, was used to determine red blood cell zinc concentrations in more than 200 samples obtained from the Neonatal Intensive care Unit at the Children's Hospital of the King's Daughters.

This study demonstrated that erythrocyte zinc levels in premature infants were similar to those of healthy adults. A linear relation between plasma and red blood cell zinc concentrations was …

Direct Optical Scanning Gel Chromatography: Improvements To The Technique And Application To Several Diverse Biochemical Problems, John Grothusen

All Dissertations

No abstract provided.

A Study Of The Action Of Bovine Cathepsin D On Intramuscular Connective Tissue, Elimosaria Elikalia Maeda

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Cathepsin D has been postulated to play a role in the tenderization of meat with ageing. Meat toughness has been related to connective tissue content and degree of myofibrillar shortening. This investigation was undertaken to determine the activity of cathepsin D on bovine intramuscular connective tissue.

Highly purified cathepsin D preparations from bovine spleen and bovine sternomandibularis muscle were obtained by ammonium sulfate fractionation followed by affinity chromatography on pepstatin coupled to controlled pore size alkyl amine glass beads. Maximum proteolytic activity using hemoglobin as substrate was obtained at pH 3.5 and 3.8 for bovine spleen and muscle cathepsin D, …

Investigation Of The Substrate Binding Site Of Nad Glycohydrolase In Human Seminal Plasma, Sau C. Cheung

Chemistry & Biochemistry Theses & Dissertations

Nicotinamide adenine dinucleotide glycohydrolase (NADase) (E.C. 3.2.2.5) from human seminal plasma (HSP) was partially purified through a multistep purification procedure, including affinity chromatography on Amicon Red Matrex Gel A. The NADase activity was assayed by a fluorometric method with nicotinamide 1,N⁶ -ethenoadenine dinucleotide as the substrate. The enzymatic cleavage of etheno-NAD by NADase was accompanied by an eight-fold increase in fluorescence. This large increase in fluorescence provided a sensitive way to follow the rate of etheno-NAD hydrolysis.

NADase was found to be less specific for the purine moiety of NAD, since etheno-NAD was accepted as a substrate. The enzyme …

Conformational Analysis Of Nonsteroidal Estrogen Analogs, Robert N. Spady

Loma Linda University Electronic Theses, Dissertations & Projects

In target tissue, estrogens initiate a series of events, ending in protein synthesis, by binding to a cytoplasmic receptor. Once the estrogen binds, the receptor complex is activated--a process including dimerization of the estrogen complex with another subunit. In order to study the possible receptor-ligand interactions, conformational analysis was performed on several nonsteroidal estrogen analogs, using the CAMSEQ8 molecular mechanics program.

The probability that a molecule will exist in a given conformational energy well depends both on the energy and the shape of the well. A method has been previously worked out to calculate probabilities using both of these variables, …

Acetylation Of Synaptosomal Protein: Inhibition Of Veratridine / Soll Berl, Ramiro Nunez, Arlene D. Colon, And Donald D. Clarke Mount Sinai School Of Medicine, And Chemistry Department, Fordham University, New York, New York, U.S.A., Soll Berl, Ramiro Nunez, Arlene D. Colon, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Incubation of synaptosomes with [3H]acetate results in rapid labeling of protein. Labeling is decreased in the presence of veratridine, and the effect of veratridine is blocked by tetrodotoxin. Most of the radioactivity can be removed by base or acid hydrolysis, and is probably incorporated as acetate; it is this fraction that is affected by the veratridine. The data suggest that veratridine stimulates deacetylation of synaptosomal protein. This raises the question whether acetylation-deacetylation is involved in membrane function

The Metabolic Compartmentation Concept / S. Berl And D. D. Clarke Mt. Sinai School Of Medicine, Bronx, N.Y. 10458; Department Of Chemistry, Fordham University, New York, N.Y. 10029., Soll Berl, Donald Dudley Clarke Phd

Chemistry Faculty Publications

The concept of metabolic compartmentation describes the presence in a tissue of functionally different and chemically distinct pools of a given substrate. These separate pools equilibrate only very slowlyt if at a11, and exhibit different turnover and flux rates. Such heterogeneous functional pools of amino acids were coming under investigation in microorganisms (Britten et al. 1955; Cowie, Walton 1956; Cowie, McClure 1959), plants (Steward et al. 1956; Maclennan et al. 1963), and animal tissues (Korner, Tarver 1957; Green, Lowther 1959; Kipnis et al. 1961) at about the same time that we began our studies on glutamate-glutamine metabolism in brain. The …

Fluoroacetate As A Possible Marker For Glial Metabolism In Vivo / Donald D. Clarke, Ph.D. And Soll Berl, M. D. Dept. Of Chemistry, Fordham Univ. And Dept. Of Neurology, Mt. Sinai School Of Medicine Bronx, N.Y. 10458 And New York, N.Y. 10029, Donald Dudley Clarke Phd, Soll Berl, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Glucose is the major source of energy in all brain cells and its metabolism is closely coupled to functional activity. Advantage has been taken of this by Sokoloff (1977) and Sokoloff et al. (1977) to study energy metabolism in different brain areas and their response to a variety of drugs and stimuli. For this purpose these workers have developed the procedure for mrasuring autoradiographically the accumulation of [14C]-deoxyglucose into its phosphate derivative, a compound which does not differentiate glial from neuronal metabolism. They could be enhanced if one could differentiate glial from neuronal metabolic response to the various experimental conditions. …

Water Quality Trends In The Las Vegas Wash Wetlands, F. A. Morris, L. J. Paulson

Publications (WR)

The Las Vegas Wash is a wetlands ecosystem that acts to buffer the effects of wastewater discharges on the receiving waters of Lake Mead. The wash is the terminus for the 4,144 km2 Las Vegas Valley drainage basin, emptying into Las Vegas Bay of Lake Mead (Colorado River). It is in the northern Mojave desert, which receives an average of only 10 cm of rainfall annually. The Las Vegas Wash is technically an artificial wetland supported almost entirely by the perennial flows from sewage treatment plants. These flows contribute an average of 3-7 t of nutrients (nitrogen and phosphorus) and …

Biochemistry, W. E. Cornatzer

UND Departmental Histories

This departmental history was written on the occasion of the UND Centennial in 1983.