Chemicals and Drugs Commons^™

The Production And Characterization Of A Monoclonal Antibody To A Naturally Occurring Aminomalonic Acid Epitope And A Comparison With An Antibody To A Synthetically Derived Aminomalonic Acid Epitope, James L. Gulley

Loma Linda University Electronic Theses, Dissertations & Projects

The goal for this project is to undertake the first step in the study of blocking metastasis by a monoclonal antibody. The long range goal of this line of research is to study a novel mechanism for the blocking of tumor metastasis in general and prostatic cancer metastasis in particular. The problem that this addresses is the growing number of cases of prostatic cancer in the increasingly aging male population. This cancer is not lethal unless it metastasizes, yet there is no cure for the metastatic disease. The inherent specificity of a monoclonal antibody combined with the uniqueness of the …

Effects Of Furosemide And Oleic Acid On Thyroxine Binding To Isolated Pairs Of Thyroxine Binding Serum Proteins, Deborah Hustead

Loma Linda University Electronic Theses, Dissertations & Projects

Dosage dependent binding inhibition and redistribution of thyroxine (T₄) between three pairs of thyroxine binding serum proteins, TBG-TTR, TBG-albumin and TTR-albumin, by furosemide (0.0 to 1.0 mol/L) and oleic acid (0.0 to 0.10 mol/L) were studied using equilibrium dialysis with purified binding proteins at concentrations 1/200 of levels in normal human sera, separated across the dialysis membrane and competing for T₄.

Furosemide (1.0 mol/L) reduced TBG bound T₄ in both the TBG-TTR system and TBG-albumin system (>90% decrease) and inhibited T₄ binding to TTR in the TTR-albumin system (68% decrease) and to albumin …

Cerebral Microcirculatory Effects Of Maturation, Scott R. Elliott

Loma Linda University Electronic Theses, Dissertations & Projects

The present studies demonstrate that in the second (2B) and fourth (4B) branches of newborn and adult sheep middle cerebral arteries, 5-HT2 serotonergic receptors mediate contractile responses to serotonin and that alpha-1 adrenergic receptors mediate contractile responses to norepinephrine. In addition, tissue sensitivity to 5-HT and NE decreases significantly with maturation, but does not vary with branch order. These age-related changes were associated with a decrease in affinity and maximum response in 2B and 4B segments of 5-HT and NE contracted arteries, respectively. Norepinephrine and 5-HT occupancy at the pD₂ rose significantly in 2B segments, but did not change …

Signal Transduction In Bacterial Chemotaxis, Edward Heath Rowsell

Loma Linda University Electronic Theses, Dissertations & Projects

This study investigated the site of ATP utilization in the signal transduction pathway of bacterial chemotaxis and localized the point of convergence of a methylation-independent system of chemotaxis with the methylation-dependent system. The identity of the signal originating from the phosphotransferase system was investigated by substituting the fructose-inducible HPr-like protein FPr for HPr in transport and chemotaxis. In addition, a novel chemoattractant, glycerol, was identified for Salmonella typhimurium. Histidine-auxotrophic S. typhimurium strains ST23 (hisF) and ST171 (hisF cheB) were depleted for ATP. The times required for the bacteria to adapt to a step increase in serine, …

A Physical Map Of The Klebsiella Pneumoniae M5a1 Genome, Alexandrine Randriamahefa

Loma Linda University Electronic Theses, Dissertations & Projects

Klebsiella pneumoniae is an enteric opportunistic pathogen which commonly causes pulmonary, urinary tract and kidney infections. It has also been widely used for the study of nitrogen fixation. In this study, a macrorestriction map of K. pneumoniae M5a1 chromosomal DNA was constructed using pulsed field gel electrophoresis (PFGE) and Southern hybridization techniques. The genome DNA was digested with two rare cutting restriction enzymes (XbaI and BlnI) and the genome size was estimated from the sum of the fragment sizes as 5,121 Kb (+2.8%). A tentative order of fragments was deduced using Southern hybridization with purified restriction fragments. …

Sds Non-Acrylamide Polymeric Gel-Filled Capillary Electrophoresis For Molecular Size Separation Of Protein, Devon Andres

Honors Theses

Sodium dodecyl sulfide (SDS) non-acrylamide gel-filled capillary columns are a new technology being used for analysis and separation of biotechnology-derived proteins. This research was to compare this new technology to the current methods of SDS polyacrylamide gel electrophoresis (SDS-PAGE) and high-performance size-exclusion chromatography (HPSEC). The molecular mass of four different recombinant proteins were determined by two commercialized SDS non-acrylamide gel-filled capillary columns, SDS-PAGE, and HPSEC. The data obtained showed that the SDS non-acrylamide gel-filled capillary columns were compatible with the SDS-PAGE technique for molecular mass determination. HPSEC was shown to be unreliable for molecular weight determination. SDS non-acrylamide gel-filled capillary …

Aerotaxis In Bacillus Subtilis, Laurence Samuel Wong

Loma Linda University Electronic Theses, Dissertations & Projects

B. subtilis, like Escherichia coli and Salmonella typhimurium, responded to a step increase in oxygen concentration by swimming smoothly (~ 35 s duration) and to a decrease in oxygen by tumbling. In a spatial gradient of oxygen in liquid media, a band of cells congregated near the air interface. Aerotaxis required a functioning respiratory chain. Adaptation of B. subtilis, E. coli and S. typhimurium to media containing amino acids requires methylation of receptors/methyl-accepting chemotaxis proteins. Adaptation to oxygen and phosphotransferase sugar substrates in B. subtilis was dependent on methylation, unlike similar adaptation in E. coli and S. typhimurium. …

The Isolation And Identification Of Polyphenolics From Grape Seeds And Their Activity Towards Specific Cell Lines, Joseph A. Reddy

Chemistry & Biochemistry Theses & Dissertations

Grape seed extract is being marketed in France for cardiovascular use and skin care. The active constituents of both these drugs are certain polyphenols called procyanidins. A similar seed extract has now been obtained from Virginia Chardonnay grape seeds. Ten of the major components in this extract have been isolated in their pure form by the semi-preparative HPLC separation on a YMC ODS-AQ (Octadecylsilane-AQ) column. The ODS-Aq column has been shown to have a better resolving capacity compared to the Spherisorb ODS (Octadecylsilane) column previously described in the literature. This has been attributed to the ODSAQ support which has both …

Longitudinal Study Of Plasma And Erythrocyte Selenium Levels In Type I Diabetic Children During Development And Treatment Of Ketoacidosis, Pi-Hsia Fan

Chemistry & Biochemistry Theses & Dissertations

Most studies on the biological activity of selenium have focused on its role at the active site in the ubiquitous enzyme, glutathione peroxidase (GSH-Px) which catalyzes the removal of hydrogen peroxide and organic hydroperoxides (1). A connection between selenium deficiency and a wide range of diseases in animals and humans has been reviewed (2). Development of various forms of human cancer (3), failure of the immune system (4), as well as some cardiovascular diseases (5) have been reported or suspected to correlate with selenium deficiency. However, the physiological function of selenium in these different diseases is not fully understood, and …

Separation And Quantification Of Queuine-Modified And Unmodified Transfer Rna Isoacceptors, Jorge Miguel Pereira De Oliveira Da Silva Santos

Chemistry & Biochemistry Theses & Dissertations

The discovery of a base-exchange modification in the first position of the anticodon of four transfer RNA (tRNA) isoacceptor families has been shown to be correlated with a decrease in the rate of cell division in several tumorigenic tissues. This fact led to the hypothesis that wobbling in the anticodon might be a control point wich mediates the rate of translation. A suitable method for separation of different tRNA isoacceptor families had to be created in order to generate a means for quantitatively measuring degrees of this base modification in tRNA. Radio labelled amino acids were used to charge tRNA …

Choosing Shower Dips For Sheep Lice, Tony Higgs, Bob Love

Journal of the Department of Agriculture, Western Australia, Series 4

The performance of all shower dip chemicals for sheep lice can vary depending on the active chemical and the conditions under which they are used.

Department of Agriculture trials have indicated that the wettable powders coumaphos and magnesium fluorosilicate were less effective at eradicating sheep lice than were synthetic pyrethroid and organophosphate dipping chemicals.

However, failure to eradicate sheep lice may be the result of several factors, many of them related to management.

Kinetic Characterization Of A Recombinant C-Terminal Mutant Of Reverse Transcriptase From The Human Immunodeficiency Virus, Thomas S. Heard

Chemistry & Biochemistry Theses & Dissertations

The human immunodeficiency virus (HIV) reverse transcriptase (RT) (EC 2.7.7.49) is the central replication enzyme for HIV. In general, the kinetic mechanism for this and all other polymerases involves the ordered binding of two substrates: a primer-template (PT) followed by a deoxyribonucleoside triphosphate (dNTP). Previous investigations prompted this research when it was discovered that the substrate dNTP, in absence of PT, could protect a recombinant c-terminal mutant HIV-1 RT from inhibition by pyridoxal-5'-monophosphate (PLP), an active-site dNTP inhibitor. In contrast, the non-mutant recombinant HIV-1 RT required both substrates for protection from PLP inhibition. This investigation sought to determine if this …

Book Review, Kristin S. Shrader-Frechette

RISK: Health, Safety & Environment (1990-2002)

Review of the following: ELAINE DRAPER, Risky BUSINESS: GENETIC TESTING AND EXCLUSIONARY PRACTICES IN THE HAZARDOUS WORKPLACE. (Cambridge University Press 1991) [315 pp.] Index of names and subjects, glossary, notes, references. LC 90-28112; ISBN 0-521-37027-2 (cloth $49.50); ISBN 0-42248-5 (paper $15.95). [40 W. 20th St., New York NY 10011.]

Plasticity Of The Gnrh Neuronal System During Sexual Maturation Of The Male Djungarian Hamster, Kevin L. Buchanan

Loma Linda University Electronic Theses, Dissertations & Projects

Onset of puberty in male Djungarian hamsters is characterized by increased pituitary gonadotropin secretion and testes maturation, and is controlled by neurons within the brain which secrete gonadotropin-releasing hormone (GnRH). Puberty is associated with increased numbers of morphologically unipolar, but not bipolar, GnRH neurons in the medial preoptic area (MPOA) and diagonal band of Broca (DBB). To test the hypothesis that delayed sexual maturation arrests this increase, males were exposed to short days or administered melatonin. Males with delayed puberty had significantly fewer unipolar GnRH neuron numbers in the MPOA and DBB comparable to pubertal controls in long days. Unipolar …

The Expression And Regulation Of The Hsdk Genes Of Escherichia Coli K-12, Ainu Parkash

Loma Linda University Electronic Theses, Dissertations & Projects

The DNA restriction and modification (R-M) genes in bacteria play a defensive role in the organism, offering protection to the host from invaders such as bacteriophages. The expression of these genes in E. coli K-12 had not previously been studied due to the complexity of the EcoK R-M enzymes. The use of lacZ gene fusion techniques allowed studies of the expression of the hsdK genes for the first time. Operon fusions of Mu(/ac) to the hsdK genes show that the two hsdK promoters, p _res and p _mod, are weak (50 Miller units) and that …

Improved Recovery Of A Radlolabeled Peptide With An Albumin-Treated Reversed-Phase Hplc Column, David S. Hage, Robert L. Taylor, Pai C. Kao

David Hage Publications

Reversed-phase high-performance liquid chromatography (RP-HPLC) is an important tool in the purification of radiolabeled peptides and proteins for immunoassay. However, for some proteins and peptides it is difficult to achieve reproducible behavior in RP-HPLC because of the low recovery of these compounds. Factors that can be varied to improve recovery include the strength or pH of the mobile phase, the chain length and spacing of groups on the reversed-phase support, and the flow rate or steepness of the elution gradient (1-5). ... In summary, we obtained better recovery and more reproducible chromatographic behavior for labeled 1-34 PTHrP with an albumin-pretreated …

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 …

High Performance Liquid Chromatographic Separation Of Different Types Of Collagen, Yongjoo Chung

Chemistry & Biochemistry Theses & Dissertations

The objective of this research was to develop a method for the separation of different types of collagen using a large pore size (100 nm) reversed-phase C-8 column. Three different types of collagen (type I, III, and V) were sufficiently separated on the column using a mobile phase containing water-acetonitrile-trifluoroacetic acid. Collagens found in each HPLC peak were identified using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to assay the column effluents.

Trifluoroacetic acid (TFA) was used as an ion pair reagent. This compound improved the chromatographic profile for these proteins on a reversed-phase column. It was observed that with …

Catecholamines And The Regulation Of Uterine Contractions In The Pregnant Rhesus Macaque, Cahleen Mae Mcnutt

Loma Linda University Electronic Theses, Dissertations & Projects

This study was designed to test the hypothesis that: 1) maternal and/or amniotic fluid catecholamines or prostaglandins display nocturnal activity rhythms similar to those of the myometrial contractility in the pregnant rhesus macaque and, 2) catecholamines stimulate prostaglandin production by fetal membranes. Paired maternal arterial blood and amniotic fluid samples were collected at 0900 h (AM) and 2100 h (PM) from chronically catheterized rhesus macaques until delivery and analyzed for prostaglandin metabolites (PGFM and PGEM-II) and catecholamines (norepinephrine, epinephrine, and dopamine). Uterine activity data was continuously recorded. A rhythm in uterine contractility was observed, with peak activity between 1900 and …

The Effects Of Glutathione On Human Keratinocyte, Manisha N. Trivedi

Theses and Dissertations in Biomedical Sciences

Cultured human keratinocytes were grown in vitro to assess the effects of glutathione, a thiol, on human keratinocyte proliferation. Single keratinocytes were harvested from newborn (greater than 5 days old) foreskin and were grown in culture flasks such that experiments could be performed in approximately 2 to 3 weeks. In order to evaluate the effects of glutathione (GSH) on keratinocyte growth, experiments were performed to determine the growth curve, plating efficiency, and incorporation of radiolabelled thymidine. Growth studies, performed with different concentrations of glutathione, revealed that 0.065 μM was the most effective concentration when compared to control cell cultures, and …

Activation In Vitro Of Transfer Rna-Guanine Ribosyltransferase By Protein Kinase C, Panayota Eriotou

Chemistry & Biochemistry Theses & Dissertations

The purpose of this study was to isolate and purify the enzymes, transfer RNA-guanine ribosyltrasferase and protein kinase C, and to determine whether the phosphorylating enzyme activates the insertion enzyme in vitro. Transfer RNA-guanine ribosyltransferase lost activity within several days after isolation and total, complete reactivation was accomplished in the presence of protein kinase C. This demonstrated that ribosyltransferase's instability is related to the degree of its phosphorylation. It is proposed that modification of tRNA is controlled by protein kinase C. Deactivation of the insertion enzyme leads to hypomodified tRNA which in turn is associated with neoplasia. Potential use …

Assessment Of Lipid Extraction From Human Bone On Subsequent Demineralization, Jaeeun Lee

Biological Sciences Theses & Dissertations

The purpose of this study was to assess the effectiveness of various solvents m removal of lipids from human bone, and to determine the effects of lipid extraction on subsequent acid solubilization of the mineral components of bone. Specifically, the following processing steps were studied; 1) ground human bone was de fatted in various ethanol / water mixtures, and / or chloroform / methanol ( 2: 1, v:v ), 2) ground human bone which had lipids extracted ( in absolute ethanol ) and bone which had not had lipids extracted were demineralized using vanous concentrations of hydrochloric acid to assess …

Laminin Potentiates Differentiation Of Pcc4uva Embryonal Carcinoma Into Neurons, T. M. Sweeney, Roy C. Ogle, C. D. Little

School of Medical Diagnostics & Translational Sciences Publications

The embryonal carcinoma PCC4uva differentiates into neurons in response to treatment with retinoic acid and dbcAMP. We used this in vitro model system to study the effects of laminin on early neural differentiation. Laminin substrata markedly potentiate neural differentiation of retinoic acid and dbcAMP-treated cultures. Only laminin induced more rapid neural cell body clustering, neurite growth and neurite fasciculation as compared to type IV collagen, type I collagen, and fibronectin substrata. Exogenous laminin substrata promoted greater cell attachment, cellular spreading and growth to confluence than type IV collagen, type I collagen, fibronectin and glass substrata. Laminin-induced effects were inhibited by …

The Determination Of Clomiphene Citrate And Tamoxifen In Human Blood Plasma And Semen, Janice Irene Kiss

Chemistry & Biochemistry Theses & Dissertations

Triphenylethylene antiestrogen compounds are used to stimulate ovulation in anovulary women and spermatogenesis in oligozoospermatic males. Clomiphene Citrate (CC) is given therapeuticaly as a mixture of the trans (E) and cis (Z) isomers. Each of these isomers has its own characteristic agonist-antagonist profiles. Only the trans form of Tamoxifen (T) is administered therapeutically.

HPLC methodologies were developed for the determination of CC and Tin human blood and seminal plasma. The methodologies involved extraction of the triphenylethylene compounds using hexane, followed by evaporation of the hexane under N₂. The residue was then taken up in a volume of HPLC …

Microwell Based Solid Phase Competitive Enzyme Immunoassay For Estradiol And Estriol, Jeng-Ting Yang

Chemistry & Biochemistry Theses & Dissertations

A sensitive and simple microwell based competition enzyme immunoassay for the quantitative determination of estradiol (E₂) and estriol (E₃) was developed. Nicrowells coated with antibody were incubated with antigen followed by adding horseradish peroxidase (HRPO) conjugate. The assay, which can be performed within two hours at room temperature, involved simultaneous incubation of E₂ - or E₃-HRPO conjugate and serum sample in polystyrene microwells coated with anti- E₂ or anti-E₃ gamma globulin fraction.

Gamma globulin was isolated from whole anti-serum by DEAE-cellulose chromatography. A carbodiimide coupling method was utilized to prepare the …

Development Of A Solid Phase Enzyme Immunoassay For Total Serum Testosterone, Li Li

Chemistry & Biochemistry Theses & Dissertations

An enzyme-labeled immunoassay of sufficient range and sensitivity was developed for serum testosterone determination. In this method, the testosterone in the serum sample and testosterone-HRPO conjugate compete with each other for the limited antibody binding sites on the anti-testosterone gamma globulin coated microwell. The assay can be performed in only 2.5 hours.

The gamma globulin fraction was isolated from the anti-testosterone whole serum by DEAE-cellulose column chromatography. Testosterone-peroxidase conjugate was prepared by a carbodiimide coupling method. The interference which arises from the binding of testosterone to steroid hormone binding protein was eliminated by the addition of 17 beta-estradiol. All assay …

Effect Of Glucose Administration On Hamster Liver S9-Mediated Mutagenesis, Metabolism And Dna-Binding Of Benzo[A]Pyrene And Aflatoxin B1, Daniel C. Rubano

Loma Linda University Electronic Theses, Dissertations & Projects

Hamster liver S9 prepared from control animals and animals given 30% glucose in drinking water 48 h before time of sacrifice was used in studies of benzo[a]pyrene (BaP) and aflatoxin (AFB₁)-induced mutagenesis, metabolism of BaP and AFB₁, and metabolite binding to calf thymus DNA. BaP-induced mutagenesis in Salmonella typhimurium TA100 was reduced 38.5% while AFB₁-induced mutagenesis was increased 36% by S9 from glucose-treated hamsters. The reduction of [³H]BaP metabolite binding to calf thymus DNA in incubations with S9 from glucose-treated hamsters correlated with a decrease in unknown BaP metabolite-deoxyribonucleoside adducts isolated by …

Effects Of Sodium Intake, Furosemide, And Infusion Of Atrial Natriuretic Peptide On The Urinary And Metabolic Clearances Of Arginine Vasopressin In Normal Subjects, Arnold M. Moses, Carol Jones, Carolyn B. Yucha

Nursing Faculty Research

Arginine vasopressin (AVP) and atrial natriuretic peptide (ANP) have important influences on water and electrolyte metabolism, and studies on the interactions between these hormones may have important implications. We have investigated the effects of sodium intake, furosemide, and infusion of ANP on the urinary and metabolic (nonurinary) clearances of AVP in hydrated normal subjects. On a high sodium diet there was an increase in urine volume, sodium excretion, osmolal clearance, plasma ANP concentration, and urinary clearance and fractional excretion of AVP, with a decrease in PRA. The infusion of furosemide increased urine volume, sodium excretion, osmolal clearance, and PRA, but …

The Accumulation And Run-Down Of Dieldrin In Wethers Grazed On Paddocks Previously Treated With Dieldrin, Tony Albertsen, Roy Casey, Keith Croker

Journal of the Department of Agriculture, Western Australia, Series 4

In mid 1987, the organochlorines (O/Cs) DDT, dieldrin, heptachlor and chlordane were deregistered for agricultural and horticultural use in Australia. These chemicals had been used widely, and land on several hundred Western Australian farms, particularly where potatoes had been grown, was contaminated with chemical residues. Livestock grazing pastures on such land was also contaminated, and our beef exports were threatened. In 1988, the Western Australian Department of Agriculture started a two-year project to collect information from sheep which were grazed on land that had previously been treated with O/Cs, particularly dieldrin. The levels of O/Cs in body and wool fats …

Effect Of Disulfiram On The Urinary Excretion Of 5-Hydroxytryptophol And 5-Hydroxyindoleacetic Acid In Subjects Consuming Serotonin Rich Foods, Amy G. Tordecilla

Loma Linda University Electronic Theses, Dissertations & Projects

The effect of disulfiram and/or serotonin rich foods (SRF) on the urinary excretion of 5-hydroxytryptophol (5-HTOL) and 5-hydroxyindoleacetic acid (5-HIAA) in normal male subjects was investigated. Urine collections were obtained on 1) regular serotonin free diet, 2) SRF, 3) serotonin free diet with disulfiram, and 4) SRF with disulfiram. Urinary excretion of 5-HTOL in ug/gm creatinine during disulfiram administration was 112.0 ± 24.1. Five HTOL (ug/gm creatinine) with disulfiram and SRF increased to 1532 ± 352.0 (p < 0.001). Five-HIAA (mg/g creatinine) before and after SRF was 3.4 ± .44 and 33.1 ± 6.1 respectively, p < 0.001. With disulfiram and SRF the excretion of was 24.2 ± 3.2 mg/g creatinine which was significantly greater than when on the regular diet (p < 0.001) than the excretion on SRF alone (p < 0.05)

Key words: Disulfiram, serotonin, 5-hydroxytryptophol, 5-hydroxyindoleacetic acid, urinary excretion, high performance liquid chromatography, alcoholism