Biochemistry, Biophysics, and Structural Biology Commons^™

Intact Parathyroid Hormone: Performance And Clinical Utility Of An Automated Assay Based On High-Performance Immunoaffinity Chromatography And Chemiluminescence Detection, David S. Hage, Bob Taylor, Pai C. Kao

David Hage Publications

The performance and clinical utility of an automated assay of intact parathyroid hormone (parathyrin, PTH) are evaluated. The method is based on the extraction of PTH from plasma by an HPLC column containing immobilized anti-(44-68 PTH) antibodies. The PTH retained is detected with a postcolumn reactor and use of anti-(1--34 PTH) chemiluminescent-labeled antibodies. The total cycle time of the assay is 6.5 mm per injection after a 1-h incubation.The lower limit of detection for PTH in a 66-pL plasma sample was 0.5 pmol/L based on peak heights and 0.2 pmol/L based on peak areas. Mean analytical recovery for PTH added …

Expression Of Biologically Active Human Corticosteroid Binding Globulin By Insect Cells: Acquisition Of Function Requires Glycosylation And Transport, Jayasri Ghose-Dastidar, J. B. Alexander Ross, Reza Green

Chemistry and Biochemistry Faculty Publications

Human corticosteroid binding globulin (hCBG) is a 50- to 55-kDa serum glycoprotein that binds cortisol and progesterone with high affinity. To map the steroid-binding domain and to investigate the folding pathways of hCBG, we have established an expression system based on infection of insect cells with a recombinant baculovirus encoding hCBG. Infected Spodoptera frugiperda (Sf9) cells secrete immunoreactive hCBG at high levels (16-24 pmol per 10⁶ cells per 40 h), and the recombinant protein binds cortisol with an affinity and specificity equivalent to that of human serum-derived hCBG. Thus, this system has the potential to provide large amounts of …

Use Of Affinity Chromatography In Developing Acridinium Ester-Labeled Antibodies For An Immunometric Assay Of Parathyrin, David S. Hage, Bob Taylor, Pat Schryver, Pai C. Kao

David Hage Publications

In developing an immunometric assay of intact parathyrin (parathyroid hormone, PTH), we found that affinity chromatography is a useful tool in purifying and optimizing the labeling conditions for acridinium ester-labeled antibodies. ... In summary, affinity chromatography was found to be useful in the purification of acridinium ester-labeled antibodies, particularly for removing denatured antibodies from the prepared label and for monitoring the amount of active labeled antibodies produced.

The Role Of The Voltage Gradient In The Agarose Gel Electrophoresis Of Dna, David Wheeler

Theses and Dissertations in Biomedical Sciences

In Part I of this dissertation, empirical equations for predicting DNA mobility during agarose gel electrophoresis (AGE) from voltage gradient are developed from the data of McDonnel (36) for electrophoresis in a 1.6% agarose gel. These equations represented the data well for DNA between 2 and 10 kilobase pairs (KBp) in length. A computer program, called GELSIM, which incorporates these equations is described in Part II. GELSIM was designed to allow researchers to analyze electrophoresis data by predicting the effect on DNA migration of altering the voltage of electrophoresis. In this way, electrophoretic banding patterns produced using different voltages could …

Isoprenylation Is Required For The Processing Of The Lamin A Precursor, Michael Sinensky, L. A. Beck, T. J. Hosick

Faculty Publications, Biological Sciences

The nuclear lamina proteins, prelamin A, lamin B, and a 70-kD lamina-associated protein, are posttranslationally modified by a metabolite derived from mevalonate. This modification can be inhibited by treatment with (3-R,S)-3-fluoromevalonate, demonstrating that it is isoprenoid in nature. We have examined the association between isoprenoid metabolism and processing of the lamin A precursor in human and hamster cells. Inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase by mevinolin (lovastatin) specifically depletes endogenous isoprenoid pools and inhibits the conversion of prelamin A to lamin A. Prelamin A processing is also blocked by mevalonate starvation of Mev-1, a CHO cell line auxotrophic for mevalonate. …

Estuarine Studies : An Activities Text For Maine Schools, Education Division - Department Of Marine Resources

Maine Collection

Estuarine Studies : An Activities Text for Maine Schools

Fisheries Education Unit #16 - Produced by The Education Division, Department of Marine Resources, State House Station 21, Augusta, Maine 04333-0021.

Revised 1990. Printed Under Appropriation No. 3140.3260.

Rotational Symmetries Of Nuclear States: Spin Determinations In Advanced Laboratory, Wilfred J. Braithwaite

Journal of the Arkansas Academy of Science

An advanced laboratory experiment is described which shows the connection between the rotational symmetries of nuclear states and the assignments of spins to discrete nuclear states. Standard angular correlation methods were used to study the two sequential gamma ray transitions in each ⁶⁰Ni nucleus, populated by unobserved beta decays from a weak radioactive ⁶⁰Co source. The chosen electronics and detectors were inexpensive and easy to operate. This experiment was extended to introduce students to real-world data acquisition, using finite-geometry detectors, which resulted in enormously larger coincident data rates.

Simultaneous Determination Of 3-Methoxy-4-Hydroxyphenyl-Glycol, Homovanillic Acid, And 5-Hydroxyindoleacetic Acid In Plasma By High Performance Liquid Chromatography, Yung-Te Tseng

Chemistry & Biochemistry Theses & Dissertations

A rapid, reliable, and sensitive method for simultaneous determination of 3-methoxy-4-hydroxyphenylglycol (MHPG), Homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5- HIAA) in plasma is described. The method is based on high performance liquid chromatography with electrochemical detection. Samples were filtered with Amicon Centrifree micropartition systems and extracted with organic solvent before chromatography. Chromatographic separation of sample components was achieved isocratically in a C18 reverse-phase column. Quantitation was performed by measuring the peak height ratios of the sample components to an internal standard. Analytical recoveries ranged from 50.78 (SD 4.2%) to 67.94 (SD 3.28). Within-run coefficient of variations (CVs) ranged from 3.78 …

A Study Of Denitrosation Of N-Nitroso Compounds By Irradiation With Long-Wavelength Uv Light, Zhenyu J. Wang

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

N-Nitrosamines and N-nitrosamides have been reported to be mutagenic. N-Nitrosamides are direct mutagens that need no activation to be mutagenic, whereas N-nitrosamines need to be enzymatically activated to exert their mutagenicity. Oxidative demethylation of nitrosamines is a commonly accepted activation mechanism. Another pathway of nitroso compounds, denitrosation, has recently been proposed. The mechanism of denitrosation, however, is still unknown.

The purpose of this study was to use a photo -reaction model to explore the possible denitrosation mechanism of N-nitroso compounds. An N-nitrosamine, N-nitrosomorpholi ne (NMOR), and an N-nitrosamide, N-methyl-N'-nitrosoguanidine (MMNG), were irradiated with long-wavelength UV light in the presence of …

Limnological Monitoring Data For Lake Mead During 1988, Suzanne E. Leavitt, Larry J. Paulson, State Of Nevada: Division Of Environmental Protection

Publications (WR)

Limnological monitoring was conducted in Las Vegas Bay and Boulder Basin from April to December of 1988. The purpose of the monitoring was to (i) document possible changes in water quality resulting from decreased phosphorus loading and increased ammonia in Las Vegas Wash, and (ii) establish a data base for evaluating the adequacy of water quality standards.

Identification Of A Variant Form Of The Human Estrogen Receptor With An Amino Acid Replacement, T. Garcia, M. Sanchez, J. L. Cox, P. A. Shaw, J. B. A. Ross, S. Lehrer, B. Schachter

Chemistry and Biochemistry Faculty Publications

Estrogen receptor (ER) is a ligand-dependent transcriptional regulator. ER protein has several functionally distinct regions, including a DNA-binding domain in its midportion, a ligand binding domain in the carboxy terminal region, and at least one domain, in the amino terminal region, that influences the magnitude of transcriptional enhancement (1,2). We now report the sequence of a frequently occurring ER genetic variant that carries two point mutations in the region of the gene encoding the amino terminal domain of the protein.

The Synthesis And Evaluation Of 5-Phenyloxazolidines As Potential Cardiovascular Drugs, Tammy C. Wang

Chemistry & Biochemistry Theses & Dissertations

Drugs such as adrenoceptor antagonists and certain centrally acting agents are known to lower blood pressure and thus serve as clinically useful antihypertensive agents. Norepinephrine 4, a natural ∝-adrenoceptor agonist, is the major agent associated with increased blood pressure in the periphery. Based on a careful structure-activity comparison, a series of substituted oxazolidines, 20, 21, 22 and 23, have been designed as possible antihypertensive candidates. Specifically, 22 and 23 have been synthesized to serve as potential norepinephrine antagonists.

Preliminary evaluation of these compounds indicated that they were not antihypertensive in nature. The more active compound, 23, …

Decreased Triiodothyronine Binding To The Hepatic Nuclear Thyroid Hormone Receptor In The Diabetic Mouse, Thomas J. Dewind

Loma Linda University Electronic Theses, Dissertations & Projects

The diabetic C57BL/KsJ-db m mouse has abnormal thyroid hormone levels and indications of thyroid hormone resistance. To investigate the basis of these abnormalities, the hepatic nuclear thyroid hormone receptor was extracted with 0.4 M KCl, 1.1 mM MgCl₂, 20 mM Tris/HCl, pH 7.9 from hepatocyte nuclei of normal C57BL/KsJ, heterozygous C57BL/KsJ-db m (db/m), and diabetic C57BL/KsJ-db m (db/db) mice. Normal and heterozygous mice were grouped together as the controls. Triiodothyronine (T₃) binding studies at 4°C using nitrocellulose filtration to separate free T₃ from receptor bound T₃ demonstrated an …

Characterization Of The Aquatic Environment In Lake Mead Near The Proposed Spring Canyon Pumped-Storage Project, And Assessment Of Potential Aquatic Impacts, Charles R. Liston, Stephen J. Grabowski, Bureau Of Reclamation

Publications (WR)

A pumped storage system consists of an upper reservoir and lower reservoir separated by an elevation difference. During low demand energy periods such as nights and weekends water is pumped from the lower to the upper reservoir using available energy from conventional steam electric power plants. During high energy demand periods, such as mornings and afternoons of weekdays, upper reservoir water is allowed to drop back down through the same system of water conduits and turbines, generating electricity to conveniently meet abrupt electrical energy requirements. The same water turbines thus act both as pumps and as conventional hydroelectric turbines. Because …

Use Of 52Cr For The Quantitative Determination Of Red Cell Volume, Claude Michael Hanbury

Chemistry & Biochemistry Theses & Dissertations

Chromium-52 has recently been suggested for use as a new agent to label red cells for the in vivo quantitation of red cell volume (1). In this paper, the development and validation of a routine ⁵²cr labeling procedure is described.

The accuracy, precision, and detection limits of chromium analysis by Zeeman effect atomic absorption spectroscopy was evaluated in the concentration range of 1 - 10 ug Cr/L.

Red cell chromium uptake was evaluated as a function of time, temperature, and concentration. Red cells labeled with a 2.5 mg/L chromium solution for 30 minutes at room temperature exhibited optimal label …

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 …

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 …

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 …

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.

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 …

Mössbauer Spectroscopic Studies Of Hemoglobin And Its Isolated Subunits, G. R. Hoy, D. C. Cook, R. L. Berger, F. K. Friedman

Physics Faculty Publications

Samples of 90% enriched ⁵⁷Fe hemoglobin and its isolated subunits have been prepared. Mössbauer spectroscopic measurements have been made on three such samples. Sample one contained contributions of oxyhemoglobin, deoxyhemoglobin, and carbonmonoxyhemoglobin. This sample was studied from a temperature of 90 K down to 230 mK. Measurements were also made at 4.2 K using a small applied magnetic field of 1.0 T. In general, the measured quadrupole splittings and isomer shifts for each component agreed with previous measurements on single component samples in the literature, and thus demonstrated that chemically enriched hemoglobin has not been altered. The second and …

Magnesium, Copper And Zinc Concentrations In Blood From Children With Diabetes Mellitus, Linda Lowers Jenkins

Chemistry & Biochemistry Theses & Dissertations

Trace element levels in blood from diabetic children were investigated and compared to control children. Plasma, erythrocyte, mononuclear and polymorphonuclear cells were analyzed for magnesium, copper, and zinc content. Leukocyte separation was performed by density gradient centrifugation on Ficoll-Hypaque, followed by washing steps to remove platelet contamination and hypotonic lysis to remove red cell contamination. Polarized Zeeman-effect atomic absorption spectroscopy was used for trace metal analysis.

Decreased plasma magnesium levels were observed in the diabetic children. This finding is well documented and probably due to increased urinary magnesium loss from glycosuria. Lymphocyte magnesium levels in the diabetic children were also …

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

Bridgewater Review

No abstract provided.

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 …