Biochemistry, Biophysics, and Structural Biology Commons^™

Reactions Of Aqueous Chlorine In The Gastric Fluid Of The Rat, Michael Gary Nickelsen

Chemistry & Biochemistry Theses & Dissertations

Gastric fluid was recovered from Sprague-Dawley rats that had been fasted for 24 hr or 48 hr and administered 4 mL deionized water. The protein, carbohydrate and chloride ion content of the gastric fluid was determined.

When gastric fluid is chlorinated with radiolabeled chlorine to concentrations between 323 and 375 mg/L, analyzed by high performance chromatography followed by liquid scintillation counting, as much as 59% of the radiolabeled chlorine was associated with organic components of the gastric fluid.

Model studies with the aromatic amino acids and the protein pepsin showed the formation of chlorinated analogs. Two chlorinated amino acid derivatives …

Studies Of Transketolase Abnormality In Alzheimer's Disease / Kwan-Fu Rex Sheu, Phd; Donald D. Clarke, Phd; Young-Tai Kim, Phd; John P. Blass, Md, Phd; Bradford J. Harding; Joseph Decicco, Kwan-Fu Rex Sheu, Donald Dudley Clarke Phd, Young-Tai Kim, John P. Blass

Chemistry Faculty Publications

The partially purified transketolase from each of eight well-nourished patients with Alzheimer's disease contained significantly less heat-stable component with a significantly longer half-life of heat inactivation than that from eight controls. Immunochemical studies utilizing antibodies to the purified human liver transketolase did not distinguish between red blood cell transketolases of patients with Alzheimer's disease and those of controls. However, three brains from patients with Alzheimer's disease that were deficient in transketolase activity lacked a 69-kilodalton form on immunoblots. Subtle structural abnormalities of transketolase appear to occur in a high proportion of patients with Alzheimer's disease

The Purification Of Ribosomal Rna Gene Chromatin From Physarum Polycephalum, Sally A. Amero, Roy C. Ogle, John L. Keating, Vicky L. Montoya, Wendy L. Murdoch, Robert M. Grainger

School of Medical Diagnostics & Translational Sciences Publications

We have undertaken the purification of ribosomal RNA gene (rDNA) chromatin from the slime mold Physarum polycephalum, in order to study its chromatin structure. In this organism rDNA exists in nucleoli as highly repeated minichromosomes, and one can obtain crude chromatin fractions highly enriched in rDNA from isolated nucleoli. We first developed a nucleolar isolation method utilizing polyamines as stabilization agents that results in a chromatin fraction containing far more protein than is obtained by the more commonly used divalent cation isolation methods. The latter method appears to result in extensive histone loss during chromatin isolations. Two methods were then …

Nucleotide Sequence Of The Fadr Gene, A Multifunctional Regulator Of Fatty Acid Metabolism In Escherichia Coli, Concetta C. Dirusso

Department of Biochemistry: Faculty Publications

The Escherichia coli fadR gene is a multifunctional regulator of fatty acid

and acetate metabolism. In the present work the nucleotide sequence of the

1.3 kb DNA fragment which encodes FadR has been determined. The coding

sequence of the fadR gene is 714 nucleotides long and is preceded by a typical

E. coli ribosome binding site and is followed by a sequence predicted to be

sufficient for factor-independent chain termination. Primer extension experiments

demonstrated that the transcription of the fadR gene initiates with an

adenine nucleotide 33 nucleotides upstream from the predicted start of

translation. The derived fadR peptide has …

Limnological Monitoring Data For Lake Mead During 1987: Technical Report No. 20, Larry J. Paulson

Publications (WR)

Limnological monitoring was conducted in Las Vegas Bay and Boulder Basin from April to December of 1987. The purpose of the monitoring was to (i) document possible changes in water quality resulting from decreased phosphorus loading in Las Vegas Wash, and (ii) establish a data base for evaluating the adequacy of water quality standards.

Changes In The Morphometry Of Las Vegas Wash And The Impact On Water Quality, Richard A. Roline, James J. Sartoris, U.S. Bureau Of Reclamation

Publications (WR)

Las Vegas Wash, a natural wash east of Las Vegas, Nevada, carries stormwater, groundwater drainage, and sewage effluent from two sewage treatment plants to Lake Mead. Over 80 percent of the normal discharge of approximately 3.4 m³/s (120 ft³/s) consists of effluent from the City of Las Vegas and Clark County sewage treatment plants. Beginning in the 1950s, a large wetland area developed along the wash that supported waterfowl populations and contributed to some water quality transformations. Heavy rains and subsequent flooding in the area in 1983 and 1984 resulted in erosion and channelization that greatly …

Incorporation Of A Product Of Mevalonic Acid Metabolism Into Proteins Of Chinese Hamster Ovary Nuclei, Michael Sinensky, L. A. Beck, T. Hosick

Faculty Publications, Biological Sciences

We have examined the nuclear localization of isoprenylated proteins in CHO-K1 cells labeled with [14C]mevalonate. Nuclear proteins of 68, 70, and 74 kD, posttranslationally modified by an isoprenoid, are also components of a nuclear matrix-intermediate filament preparation from CHO cells. Furthermore, the 68-, 70-, and 74-kD isoprenylated polypeptides are immunoprecipitated from cell extracts with two different anti-lamin antisera. Based on exact two-dimensional comigration with lamin B, both from rat liver lamin and CHO nuclear matrix-intermediate filament preparations, and its immunoprecipitation with anti-lamin antisera, we conclude that the 68-kD isoprenylated protein found in nuclei from [14C]mevalonate-labeled CHO cells is lamin B. …

Time Delayed Protein Complementation, Diana L. Lakusta

Loma Linda University Electronic Theses, Dissertations & Projects

The time delayed complementation of plant proteins was tested in weanling Sprague-Dawley male rats by measuring growth rates as well as plasma and tissue free amino acids. Diets of pinto beans with rice, corn or wheat were pair-fed in combination or alternating 4 times a day at 6 and 11 am, and 4 and 9 pm for 4 weeks. Plasma, liver and muscle samples were obtained 2 hours postprandial, a time determined most suitable for sacrifice, and analyzed for free amino acids via ion-exchange chromatography. There were no significant differences in growth of rats fed combination or alternating diets.during week …

The Refined Crystal Structure Of A Fully Active Semisynthetic Ribonuclease At 1.8 Å Resolution, Philip D. Martin, Marilynn S. Doscher, Brian Fp Edwards

Biochemistry and Molecular Biology Faculty Publications

A fully active, semisynthetic analog of bovine ribonuclease A, comprised of residues 1-118 of the molecule in a noncovalent complex with the synthetic peptide analog of residues 111-124, has been crystallized in space group P3(2)21 from a solution of 1.3 M ammonium sulfate and 3.0 M cesium chloride at pH 5.2. The crystallographic structure was determined by rotation and translation searches utilizing the coordinates for ribonuclease A reported by Wlodawer and Sjolin (Wlodawer, A., and Sjolin, L. (1983) Biochemistry 22, 2720-2728) and has been refined at 1.8-A resolution to an agreement factor of 0.204. Most of the structure of the …

Synthesis And Characterization Of N-Alkyl-Adenosine-5'-Diphosphates, Han-Ping Huang

Chemistry & Biochemistry Theses & Dissertations

n-Alkylphosphates and adenosine derivatives have been observed to be competitive inhibitors of coenzyme molecules in many NAD⁺ requiring enzymes. In earlier studies, adenosine derivatives such as AMP, ADP and ADP-ribose were found to be coenzyme competitive inhibitors of the Lactate Dehydrogenase catalyzed reaction. However, n-alkylphosphates, to the contrary, + were found to be noncompetitive inhibitors of NAD⁺.

Furthermore, it was observed previously in Dr. Yuan's laboratory that the effect of n-octylphosphate on the LDH catalyzed reaction was changed from noncompetitive to competitive in the presence of low concentration of AMP. Rossmann et al. reported that the binding …

Investigation Of Complex Formation By Oligomers Of Cytosine And Guanosine, Steven Roberts Davis

Chemistry & Biochemistry Theses & Dissertations

Duplex formation between oligo(C:G) _n where n=3 to 4 was shown not to occur under conditions favorable for duplex formation between poly G and poly C. Instead, a stable guano sine self-structure was found to form which a T_m of 50°C for (Gp)₃ and 80°C for (Gp)₄ at strand concentrations of 10^-5M in 1M NaCl. Neither a duplex nor a self-structure formed in the absence of salt.

Oligomers of guanosine and cytosine were obtained by basic hydrolysis and separated according to chain length using DEAE Sephadex column chromatography. Separation of cytosine oligomers with chain lengths …

Signal Processing In Bacterial Chemotaxis, Jimmie Marquette Smith

Loma Linda University Electronic Theses, Dissertations & Projects

This study localized 1) the site of ATP utilization in the signal transduction pathway of bacterial chemotaxis and 2) the point of convergence of the methylation-independent systems of chemotaxis with the methylation-dependent system. The site of ATP utilization was localized at the CheY protein/switch point. In an Eaeheriehia call strain (BT3077) containing the cheY, motor and switch genes, ATP depletion, accomplished by treating the cells with arsenate, suppressed the CheY protein-induced tumbling. ATP was not required by the switch for directing the clockwise and counterclockwise rotations of the flagellar motor. ATP did not appear to be required for normal steady-state …

Intracellular Protein Degradation In A Mesophile And A Thermophile Of The Genus Bacillus, Cameron C. Hoover

Masters Theses

¹⁴C-leucine was incorporated into th e proteins of the mesophile Bacillus licheniformis and the thermophile Bacillus stearothermophilus. Aliquots of the cell suspension were treated with chloramine-T in order to decarboxylate the ¹⁴C-leucine that was released as the proteins degraded. The ¹⁴CO₂ was trapped and counted. This allowed for the determination of the extent of intracellular protein degradation at various tem peratures and at various stages of the growth cun/e. For the mesophile, the extent of degradation increased with temperature from 30° C to 55° C. For the thermophile, no significant amount of degradation occurred until …

Synthesis Of Potential Long Acting Calcium Channel Antagonists: A Study Of 1,4-Dihydropyridine Analogs, Thomas E. Christos

Chemistry & Biochemistry Theses & Dissertations

An investigation was undertaken to synthesize and evaluate longer acting analogs to the antihypertensive agent nifedipine. Analogs that are capable of covalent bonding to the nifedipine receptor were considered. These compounds may show increased activity as they will remain at the receptor for a longer period of time.

During the course of this study, previously unreported and unexpected products were encountered and lead to a more intense examination of the Hantzsch synthesis. Of particular interest was the oxazolidine 25 which was a major component of the Hantzsch synthesis when ethanolamine was used instead of ammonium hydroxide. Also isolated during this …

Lake Mead Cover Enhancement Project, Jennifer Stevens Haley, Suzanne Leavitt, Larry Paulson, Donald H. Baepler

Publications (WR)

Ninety-three wildlife agencies were surveyed for information on their attempts to improve fish habitat. In addition, an annotated bibliography including over 100 summaries was completed on: 1. largemouth bass cover requirements and preferences, 2. use and effectiveness of artificial cover, 3. aquatic plant introduction and species requirements for germination and establishment, 4. terrestrial plant introduction and species requirements for germination and establishment, and 5. nutrient exchange between sediment, aquatic plants, and water. A reconnaissance of existing terrestrial and aquatic vegetation was completed in June 1986 including the production of a video tape of the Nevada shoreline of Lake Mead. Cover …

Las Vegas Wash And Lake Mead Proposed Water Quality Standards: Revisions And Rationale, State Of Nevada: Division Of Environmental Protection

Publications (WR)

Rationale of review and for proposed changes to the Nevada Pollution Control Regulations (NAC 445.1354, 445.1355, 445.1356, 455.1367, 445.1352, 445.1353, 445.1350, 445.1351) before the State Environmental Commission on June 23 and 24, 1987.

Raw Nutrient Data, Leanna Gail

Publications (WR)

Data collected at various stations around Lake Mead. Includes information about substances present in various water samples.

Purification And Characterization Of An Inhibitor Of Thymidine Uptake From Culture Supernatants Of Human Tonsil Lymphocytes, Abdolreza Zarnegar

Electronic Theses and Dissertations

Lymphocytes from human tonsils were cultured in the absence of serum for 3 days. In the presence of the concentrated culture supernatant the proliferative response of PBL, to con A, as measured by the uptake of ('3)H-tdr, was significantly reduced. The suppressor substance was referred to as SMAL (suppressor of mitogen activated lymphocytes). The estimated molecular weight of SMAL under nondenaturing conditions was 100,000-300,000. SMAL also suppressed the incorporation of ('3)H-tdr by a variety of mouse and human tumor cell lines. The activity of SMAL was sensitive to pronase and heating at 100(DEGREES)C for 30 minutes but insensitive to RNase. …

Production Of Thymine Dimer In Dna From Mesophilic And Thermophilic Bacteria By Uv Irradiation, F Redrick Shu-Chung Yein

Dissertations

The in vitro production of thymine dimer (TT) in DNA, isolated from the mesophile (B. licheniformis) and the thermophile (B. stearothermophilus), was carried out by UV irradiation (254 nm) at three different temperatures (35, 45, and 55(DEGREES)C). The production of TT from the thermophilic DNA was significantly higher than that from the mesophilic DNA. This was the case at all three irradiation temperatures. The increase in TT production per 10(DEGREES)C increase in irradiation temperature was four times higher for the thermophile than that for the mesophile. It was concluded that the DNA from the thermophile was photochemically less stable than …

Pyrimidines. 8. Chlorination Of 6-Methyluracil With Phosphorus Oxychloride In The Presence Of Trialkyamines / Herman Gershon [Ab], Anthony T. Grefig [A], And Donald D. Clarke [B] [A] Boyce Thompson Institute For Plant Research At Cornell University, Ithaca, New York 14853 [B] Department Of Chemistry, Fordham University, Bronx, New York 10458, Herman Gershon, Anthony T. Grefig, Donald Dudley Clarke Phd

Chemistry Faculty Publications

The effect of the tertiary amines triethyl, tri-n-propyl, and tri-n-butylamines on the chlorination of 6-methyluracil by phosphorus oxychloride was studied. A comparison with the reaction of preformed 2,4-dichloro-6-methylpyrimidine and triethylamine in toluene was made. The reaction in phosphorus oxychloride in the presence of triethylamine afforded low yields of 2-diethylamino derivative after short heating periods and high yields of the 2,4-bis(diethylamino) derivative after 188 hours of boiling. Heating the preformed 2,4-dichloro-6-methylpyrimidine in toluene in the presence of triethylamine yielded primarily the 2-diethylaminopyrimidine along with a small amount of the 4-diethylamino isomer. After 188 hours, the product mixture was composed of 87% …

Pyrimidines. 9. Chlorination Of 6-Trifluoromethyluracil With Phosphorus Oxychloride In The Presence Of Trialkylamines / Herman Gershon [Ab], Anthony T. Grefig [A], And Donald D. Clarke [B] [A] Boyce Thompson Institute For Plant Research At Cornell University, Ithaca, New York 14853 [B] Department Of Chemistry, Fordham University, Bronx, New York 10458, Herman Gershon, Anthony T. Grefig, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Ring-chlorination of 6-trifluoromethyluracil in phosphorus oxychloride in the presence of triethyl, tri-npropyl, and tri-n-butylamines was studied with respect to by-product formation. Comparisons were made with the results obtained by treating the preformed chlorinated pyrimidine with triethyl amine in boiling toluene. Amination of chloropyrimidines by tertiary amines takes place by a Hofmann type reaction with substituent orientation generally in the 2 position of the ring. Yields of products depended on the base and reaction time. The rate of substitution in the 2 position is significantly enhanced by the presence of the trifluoromethyl group in the 6 position as compared with a …

Acetylation Of Synaptosomal Protein: Effect Of Na+ / Arlene Colon, Soll Berl, And Donald D. Clarke, Arlene D. Colon, Soll Berl, Donald Dudley Clarke Phd

Chemistry Faculty Publications

In a previous study it was shown that the acetyl moiety can be incorporated into the protein of purified synaptosomes (1). This process was inhibited by veratridine and the inhibitory effect was counteracted by tetrodotoxin. This suggested that the flux ofNa +may be related to the acetylation process. We now report that in a sodium free medium the amount of acetylation is increased and the inhibitory effect of veratridine (veratrine) is no longer evident. The addition ofNa +leads to a decrease in acetylation in the presence of veratrine. The presence of scorpion toxin has an effect similar to that of …

Tryptidase: A Novel Trypsin-Like Enzyme From Rat Skin. Isolation And Characterization, Vincent J. Braganza

Dissertations

No abstract provided.

Role Of The Pseudomonas Aeruginosa Reca Analogue In Lysogeny And Recombination, Tyler Alan. Kokjohn

Dissertations

No abstract provided.

Spectrophotomatic Assay For The Enzyme Catalyzed Reaction Of 4-Nitroquinoline 1-Oxide With Glutathione, J. Steven Stanley, Ann M. Benson

Journal of the Arkansas Academy of Science

4-Nitroquinoline 1-oxide (4NQO(is a toxic and carcinogenic compound that has been reported to be subject to conjugation with glutathione (GSH). This reaction may proceed non-enzymatically or be catalyzed by GSH transferases. The non-enzymatic rate for this reaction has been reported to be very high. The purposes of this investigation were to develop a spectrophotometric assay for the reaction of4NQO with GSH and to determine whether the rate for the enzyme catalyzed reaction was significant relative to the non-enzymatic reaction. The absorbance spectrum of 4NQO in phosphate buffer exhibited a maximum at 365 nm. Reaction of 4NQO with GSH was accompanied …

Biochemical And Histological Evaluation Of Normal Human Epidermis Vs Cultured Human Epidermis, Ngo Thi Bich Lien

Biological Sciences Theses & Dissertations

Cultured human keratinocytes were grown in vitro for transplantation purposes in case of severe burns. Single keratinocytes were harvested from human epidermis and grown in culture flasks such that multilayered sheets of keratinocyte cells were harvestable in approximately three weeks. In order to evaluate cultured epidermis for equivalency with normal human epidermis, biochemical and histological studies were performed. Thin sections of cultured epidermis were shown to be composed of cells that had undergone developmental differentiation, but not terminal differentiation. Biochemical analyses of keratin and prekeratin proteins isolated from cultured epidermis indicated that cultured keratinocytes have electrophoretic patterns similar to keratins/prekeratins …

The Binding Of Host-Selective Toxin Analogs To Mitochondria From Normal And 'Texas' Male Sterile Cytoplasm Maize, Kurt A. Frantzen, J. M. Daly, Herman W. Knoche

Department of Biochemistry: Faculty Publications

Tritium-labeled toxin analogs were prepared by reduction with NaB³H₄ of either the toxin from Helminthosporium maydis race T or a toxin component from Phyllosticta maydis. These reduced analogs had high radiochemical specific activities, high biological activities, and plant specificities identical to the native toxins. A filtration assay was developed to test the binding of these labeled analogs to isolated- mitochondria. Binding was not energy dependent nor was there measurable matrical uptake. The analogs were shown to be lipophilic, a characteristic which gave rise to considerable nondisplaceable binding. Under conditions limiting nondisplaceable binding, -the displaceable binding was …

The Synthesis And Evaluation Of The Cardiovascular Activity Of Novel Imidazole Ring Systems, Joseph E. De Los Angeles

Chemistry & Biochemistry Theses & Dissertations

An investigation was undertaken to synthesize and evaluate the cardiovascular activity of two imidazole ring systems. Imidazoles with hypotensive activity would provide novel potential antihypertensive drugs. These novel structures may also find use as chemical probes in the study of the cardiovascular system and as prototypes for other potential cardiovascular agents.

The histamine derivative, 7,7-dipyridyl-4,5,6,7-tetrahydroimidazo-[4,5-c]pyridine (ligand l), was demonstrated to possess significant hypotensive activity in laboratory rats. A 15 mg/Kg dose of ligand 1 produces a statistically significant decrease in blood pressure when injected IP. This hypotensive effect is also associated with marked increase in heart rate. Although compound 1 …

Determination Of Human Sperm Cell Acrosin Activity From Normal And Subfertile Men, Dilrowshan H. Haque

Chemistry & Biochemistry Theses & Dissertations

Acrosin is an acrosomal enzyme which is reported to be involved in sperm penetration through cumulus, zona, and ooplasma. This enzyme has been reported to be significantly lower in sperm of infertile men compared to sperm of fertile men. The acrosin assay which has been developed can be used for studies in patients with suspected infertility and may be used on a fraction of most semen specimens being evaluated in a clinical andrology laboratory.

Within-run precision studies yielded coefficients of variation of 11% for a low-activity pool and 9% for normalactivity pool specimens. Between-run precision studies using a single donor …

Cooperativity Of Ligand Binding As A Function Of Monomer-Dimer Equilibrium Parameters And Acceptor Concentration, George K. Wolfer

Loma Linda University Electronic Theses, Dissertations & Projects

A general monomer-dimer equilibrium system involving ligand interactions is presented. Cooperativity features of specific limited models are analyzed by selecting the appropriate family of equilibrium constants from this general scheme. Each system is then characterized in terms of Hill coefficient dependency on alterations in values of equilibrium constants and total acceptor concentration. This method permits comparison of predicted cooperativity trends between systems. Contrasting reports concerning cooperativity dependencies for certain defined equilibrium systems are compared and the discrepancies resolved. Characteristics of cooperativity binding patterns are shown to include symmetry about dimerization association constant values, both positive and negative cooperativity for a …