Medicine and Health Sciences Commons^™

Regulation Of Luteinizing Hormone And Catecholamine Release, Benjamin Aaron Adler

Theses and Dissertations (ETD)

These studies tested the interrelated hypotheses that the ovarian hormones produce their positive feedback effects on luteinizing hormone (LH) secretion through activation of noradrenergic and adrenergic systems in specific hypothalamic regions. Furthermore, the ovarian hormones may alter the activity of opioid neuropeptide and Gamma-Aminobutyric Acid (GABA) systems to produce these alterations in catecholamine transmission and gonadotropin secretion. Radioimmunoassays were utilized to determine plasma LH and median eminence LHRH, and hypothalamic catecholamine concentrations were measured by radioenzymatic assay.

The first two studies tested whether epinephrine (EPI) synthesis inhibition blocks the accumulation of median eminence LHRH that precedes the ovarian hormone-induced LH …

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 …

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 …

The Effect Of Dietary Alpha - Linolenic Acid On Apparent Risk Factors Of Cardiovascular Disease, Kelvin D. Lindbeck

Loma Linda University Electronic Theses, Dissertations & Projects

Eight healthy male lacto-ovo-vegetarian subjects averaging 28.0 ± 3.6 years of age (22 to 53 years) were fed a diet rich in α-linolenic acid (18:3 ω 3) for four weeks. Arterial blood pressure and body weight were recorded each week. Plasma 6-keto-prostaglandin F_1α (6-keto- PGF_1α ), thromboxane B₂ (TXB₂), total cholesterol, triglyceride, and total phospholipid fatty acid composition were analyzed at 0, 2 and 4 weeks.

Systolic blood pressure increased significantly between weeks 0 and 1 (106.5 ± 2.7 mmHg to 114.0 ± 3.7 mmHg; p < 0.003), but decreased to 107.3 t 2.6 mrHg (p < 0.02) at week 2. There was an insignificant increase in both systolic and diastolic blood pressures after four weeks. There was no significant change in either total cholesterol or triglyceride levels during the four weeks.

Plasma 6-keto-PGF_1α levels increased significantly from 60.13 ± …

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 …

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 …

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 …

The Copper Status Of Vegetarian And Non-Vegetarian Men, Susan Bruce

Loma Linda University Electronic Theses, Dissertations & Projects

In this study, the copper status of 18 vegetarian and 17 non-vegetarian men were compared. Copper status was assessed by determining red blood cell superoxide dismutase (RBC-SOD) activity and plasma copper. Plasma copper levels were measured by atomic absorption spectroscopy. Plasma cholesterol, triacyclglycerols, HDL cholesterol, LDL cholesterol VLDL cholesterol were also determined. Twenty-four hour recalls and three-day food records were obtained to determine copper, zinc, energy, protein, fat, fiber, saturated fat, oleic acid, linoleic acid and cholesterol intake. Non-vegetarians had a significantly higher plasma cholesterol and LDL cholesterol than vegetarians. They also had a significantly higher cholesterol, saturated fat, percent …

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, …

Development Of A Procedure For Analysis Of High Density Lipoprotein Subclasses, Chan Chin

Chemistry & Biochemistry Theses & Dissertations

Human serum high density lipoprotein subclasses, HDL₂ and HDL₃ , were isolated by preparative salt density gradient ultracentrifugation and further analyzed by electrophoresis on a 4 to 15% concentration gradient polyacrylamide gel.

The separation of the major classes of serum lipoproteins was achieved after a single ultracentrifugation for 272,000 g at 15°c in a swinging bucket rotor. High resolution concentration gradient gel electrophoresis was found to be particularly suitable for the separation of lipoproteins. The isolation of HDL by precipitation methods, heparin-manganese and dextran-magnesium have also been evaluated in this study, and were found not to completely separate …

The Effect Of Hypercholesterolemia On The Membrane Lipids, 3-0-[¹⁴C] Methylglucose Transport, And 2,3-Diphosphoglycerate Levels In Rat Erythrocytes, Edwin J. Lester Jr.

Loma Linda University Electronic Theses, Dissertations & Projects

An increase in dietary cholesterol and cholic acid will produce hypercholesterolemia in rats. For ten months, two groups of rats (six in each group) were fed commercial rat pellets (control group) or rat pellets plus cholesterol (1%) and cholic acid (0.25%) (experimental group). A blood sample from each animal was divided into three aliquots for three separate experiments on the erythrocytes: a membrane cholesterol and phospholipid determination, a 3-0-[¹⁴C] methylglucose (30MG) transport determination, and a 2,3-diphosphoglycerate (2,3 DPG) determination. The 30MG transport and 2,3 DPG aliquots were each incubated in the same five concentrations of 30MG and glucose …

Muscle Catabolism During A Protein Sparing Modified Fast As Measured By Urinary 3-Methylhistidine, Judith D. Nielsen

Loma Linda University Electronic Theses, Dissertations & Projects

The urinary excretion of 3-Methylhistidine was used as an index of muscle catabolism in four groups of obese persons receiving a Protein Sparing Modified Fast (PSMF). The composition of the supplements were as follows: Group I: 75 g protein, 25 g complex carbohydrate (starch) per day; Group II: 25 g protein, 75 g complex carbohydrate (starch) per day; Group III: 75 g protein, 25 g simple carbohydrate (glucose) per day; Group IV: 25 g protein, 75 g simple carbohydrate (glucose) per day. The PSMF lasted for seven days and a 24-hour urine sample was collected at the beginning of the …

In Vitro Enzymatic Assay Of Rna Methylation, Pamela Jean Eubanks Gallup

Chemistry & Biochemistry Theses & Dissertations

An assay procedure for in vitro enzymatic methylation of mannnalian ribosomal RNA has been developed in this study. The assay procedure, utilized for the comparison of normal and neoplastic methylase activities (using mouse liver and Ehrlich ascites cells as sources of enzyme), is a modification of previously published methods (52,53). A 100,000 x g supernatant (SlOO) enzyme preparation was incubated with 28S-5.8S rRNA and tritium-labeled S-adenosyl-L-methionine. The RNA was extracted, applied to DEAE cellulose paper, washed, and the radioactivity counted. The neoplastic cell methylase preparation was more active in methylating both exogenous neoplastic and normal 28S-5.8S rRNA than the normal …

Studies Toward The Total Synthesis Of Antibiotic 593a, Robert B. Wardle

Undergraduate Honors Capstone Projects

In 1970 art article was published in the Journal of Antibiotics in which the isolation of a substance from the south African soil microorganism Streptomyces griseolutus designated 593A (NSC-135758) was reported. It was reported to inhibit the growth of human tumors in the chick embryo. Arison and Beck in 1973 proposed the structure to be 3,6-bis-(5-chloro-2 -piperidyl-)-2 ,5-piperizinedione based on spectral data.

In the ensuing years a number of further studies were made on the activity of 593A, It was found to be active against a number of solid tumors, several variants of leukemia, and in selectively inhibiting DNA synthesis. …

The Synthesis And Biochemical Evaluation Of Potential Acetylcholinesterase Reactivators, Andrea Mcdearmid Lunsford

Chemistry & Biochemistry Theses & Dissertations

Objectives of this research were to (1) synthesize, and (2) evaluate new compounds which would be potential reactivators of organophosphate-inhibited acetylcholinesterase.

A series of 1,8-diazafluorenone and 2,2'bipyridylketone oximes and oxime methiodides were synthesized (11, 13, 14, 22, 24). These compounds were developed as structural analogs of 2-PAM (2-Pyridinealdoxime methiodide) and TMB-4 (1,1Trimethylene-Bis(4-formylpyridinium bromide)dioxime), the current drugs of choice for treatment of organophosphorous poisoning.

Acetylcholinesterase from electric eel was inhibited with DFP (diisopropylfluorophosphate), an irreversible-type inhibitor, and the inhibited enzyme was treated with the various reactivator compounds. The reactivation assays were monitored with pH stat apparatus, and the data were evaluated …

Multiple Forms Of Nicotinamide Adenine Dinucleotide Glycohydrolase From Bull Semen, Stanley D. Yeatts Ii

Chemistry & Biochemistry Theses & Dissertations

Nicotinamide adenine dinucleotide glycohydrolase (NADase) of bull semen is an extracellular, soluble enzyme capable of catalyzing the hydrolysis of the nicotinamide N-ribosidic linkage of NAD⁺. In search for a rapid, simple procedure for the preparation of this enzyme, it was found that Matrex Gel Red A (Amicon) column packing served very well as an affinity gel for NADase. The enzyme was applied to the Matrex Red A column (21 cm x 1.5 cm) in 10 mM sodium phosphate buffer, pH 6.0. Surprisingly, two fractions of NADase activity were obtained when the column was eluted with a sodium chloride …

High Resolution Two-Dimensional Electrophoretic Study Of Human Seminal Plasma Proteins, Edward E. Gaunt

Chemistry & Biochemistry Theses & Dissertations

A comprehensive biochemical screening procedure for the evaluation of seminal plasma constituents is needed which, when used in conjunction with semen analysis, can provide the physician with both qualitative and quantitative information regarding the fertility status of an individual.

High resolution two-dimensional electrophoresis is a technique whereby proteins in a complex mixture such as a biological fluid are separated in one dimension by molecular charge and in the second dimension by molecular mass. It has been the purpose of this investigation to adapt and evaluate this technique for the analysis of human seminal plasma.

Our efforts have shown that this …

Lipoprotein Lipase Inhibitor Removal During Ultrafiltration Of Renal Failure Patients, Alisa Minear

Loma Linda University Electronic Theses, Dissertations & Projects

Plasma samples were obtained from seven hypertriglyceridemic (>150 mg/dl) chronic renal failure patients before and after regular hemodialysis. The effect of the plasma on the lipolytic activity of bovine milk lipoprotein lipase was investigated. The lipoprotein lipase activity, as measured by release of C¹⁴-labeled free fatty acid from triglyceride, was decreased 8-67% when incubated with uremic plasma as compared to the control plasma (44 mg/dl triglyceride). The inhibition noted prior to hemodialysis did not appear to be alleviated by the dialysis but subsequent treatment with an isolated ultrafiltration technique did appear to remove an inhibitory substance as …

Sulfatides As Opiate Receptors: An Evaluation Of The Opiate Binding Potential Of Sulfogalactosylgylcerolipid, Darby Geoghegan Hand

Chemistry & Biochemistry Theses & Dissertations

Cerebroside sulfate (CS) has recently been shown to bind opiate drugs in a pharmacologically relevent manner and therefore satisfy the structural requirements of an opiate receptor. Seminolipid (SGG), a novel sulfated galactosyl. glycerolipid that has been isolated from the testes of a variety of mammals, may be viewed as a structural analog of CS. In view of this structural similarity, it is reasonable to presume that SGG may also bind opiate drugs. The isolation of SGG from boar testes was accomplished using silica gel dry column chromotography with a chloroform: methanol water solvent system. The structure and purity of the …

Valium: Its Uses And Abuses, Jeffrey A. Hardy

Undergraduate Honors Capstone Projects

Between the months of June 1978 and June 1979, I was employed at Sunshine Terrace, a nursing home in Logan, Utah. In the year I spent there working as an orderly on the wing for incontinent male patients, I was exposed to a lot of things of which I had previously been unaware.

One which I observed was the liberal use of medications. At each nuses' station there was a small room which contained about six cabinets full of prescription drugs for the forty or so patients on that wing. At least every four hours the medication nurse would prepare …

Characteristics Of The Recovery Of The Coenzyme A-Synthesizing Protein Complex From Saccharomyces Cerevisiae, Stanley Joseph Tarnowski Jr.

Theses and Dissertations (ETD)

A multienzyme complex contained in Bakers' yeast (Saccharomyces cerevisiae) which synthesizes CoA has been named the coenzyme A-synthesizing protein complex (CoA-SPC). The CoA-SPC has been shown to be insoluble in the crude Bakers' yeast cell lysate formed by exposing the yeast cell to ether and dry ice. Only after solubilization has this multienzyme complex been shown to catalyze the formation of bound dephospho-CoA utilizing the substrates adenosine triphosphate, D-pantothenic acid and L-cysteine. A low molecular weight component or components of the soluble fraction of the yeast cell and chloride ion appears to be responsible for the solubilization of CoA-SPC. This …

Consciousness, Neurons, And Laughing Gas, Dody Michelson Orendurff

Dissertations and Theses

Psychological and physiological effects of nitrous oxide resemble those of eight other drug categories. Lipid solubility or hydrate microcrystal theories correlate behavioral measures with measurable parameters of the molecule N₂O. N₂O, a spindle poison, halts mitosis in metaphase, producing widespread physiological consequences. N₂O affects the microtubules of the spindle in a number of specific ways. Microtubules are utilized in other parts of eukaryotic cells, in a wide variety of functions. In neurons, microtubules build and maintain dendritic sensory processes. Since microtubules are built of two dissimilar proteins, constantly assemble and disassemble, and maintain a …

Isolation And Chemico-Physiologic Characterization Of A Parotid Hormone Activity From Porcine Glands, Jean-Marc Tieche

Loma Linda University Electronic Theses, Dissertations & Projects

For the past decade, evidence has accumulated which suggests that the resistance of the teeth to dental decay is in part under systemic control. It was shown that the resistance is accomplished through the optimal operation of a dentinal fluid transport mechanism which is believed to be hormonally controlled by the hypothalamic-parotid gland endocrine axis. Although circumstantial evidence for such an endocrine function was strong, the isolation of a parotid hormone which stimulates the dentinal fluid transport mechanism remained to be accomplished.

Using modern biochemical techniques, a methodology was designed to isolate and purify the hormone from porcine parotid glands. …

Human Placental Alkaline Phosphatase And Steroid Metabolism By Rat Adrenals, Allen G. Meier

Loma Linda University Electronic Theses, Dissertations & Projects

Human placental alkaline phosphatase was purified by butanol extraction, methanol precipitation and chromatography on DEAE-cellulose. The pH optimum was found to be 10.9. The enzyme was competitively inhibited by di-ethyl-p-nitrobenzyl-phosphonate and copper sulfate. An increase in enzymatic activity occurred in the presence of the phosphate ion (HPO₄) and ATP.

The enzyme was crystallized from a 40% saturated solution of ammonium sulfate and shown to be homogeneous by sedimentation analysis. Homogeneity of the enzyme was also demonstrated using poly-acrylamide-gel-electrophoresis. At a pH of 7.5% the electrophoretic pattern yielded a single band and showed no significant contamination.

The metabolic transformation …

Kinetic And Binding Studies On L-∝-Glycerophosphate Dehydrogenase, Paul Richard Rosevear

Chemistry & Biochemistry Theses & Dissertations

The properties of the coenzyme, NAD, binding site of chicken muscle L-∝-glycerophosphate dehydrogenase were studied. Adenosine monophosphate, adenosine diphosphate and adenosine diphosphoribose were sha.vn to inhibit the enzyme competitively with respect to NAD. The presence of adenosine, pyrophosphate and ribose regions in the coenzyme binding site of the enzyme was suggested by the inhibitor constants obtained for these compounds.

Disodium monoalkyl phosphates, n-butyl to n-dodecyl phosphate, inclusive, were also shown to inhibit the L-∝-glycerophosphate dehydrogenase catalized reaction competitively with respect to NAD. A positive chain length effect was observed in the binding of these compounds to the enzyme, suggesting the …

Effects Of Cariogenic And Noncariogenic Diets On The Concentration Of The Parotid Hormone Releasing Factor In Rat Hypothalami, Melva Joan Brown

Loma Linda University Electronic Theses, Dissertations & Projects

Hypothalamic and cortical extracts from various cariogenically grouped rats which had been maintained on either a Purina diet or a high sucrose diet (HSD) for three months were infused intravenously into 28 days old rats to determine if there was a titer of parotid hormone releasing factor (PRF) capable of facilitating fluid movement (FM) in the teeth. The hypothalamic and cortical extracts obtained from Purina-fed rats with 0-caries and the hypothalamic extracts from BSD-fed rats conclusively demonstrated FM stimulatory ability. Other Purina-fed groups were essentially biologically ineffective, however, extracts from HSD-fed caries groups including scores 11-40 demonstrated extremely potent FM …

The Effect Of Insulin, Fluoride And Methyl Xanthines On The Hypothalamic-Parotid Gland Endocrine Axis, Reagan Ondler

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of this study was to determine the effect of insulin and fluoride on dentinal fluid movement (FM) in high-sucrose fed rats as mediated through the hypothalamic-parotid gland endocrine axis. In conjunction with the fluoride study, the possible involvement of cAMP was examined through the use of several methyl xanthines which are recognized as phosphodiesterase inhibitors.

Twenty-one day old, male Sprague Dawley rats were maintained on a high-sugar diet (62% sucrose) for 8-9 days at which time intravenous or arterial infusions of insulin, fluoride or the methyl xanthines were performed. Additional procedures included collecting plasma from rats which had …

The Development Of A Course Of Study In Fda Drug Regulatory Procedures, Robin Adele Wills

University of the Pacific Theses and Dissertations

It is the purpose of this thesis to develop a course of study for pharmacy schools on drug regulation by the Food and Drug Administration (FDA). A summarization of the history of the development of the Food and Drug Administration (FDA) and of pharmacy education reveals a lack of interface between the two. The necessity for the interface of pharmacy education and the agency which has significant control over the products forming the basis of the pharmacy profession will be made apparent. A period of residency through which the course of study on drug regulation by the FDA was to …

The Relationship Of Alcohol Consumption In Rats To Morphine And Codeine Intake And The Metabolism Of Dopamine, Hsiu Fen Liu

Loma Linda University Electronic Theses, Dissertations & Projects

The effects of morphine and codeine free-choice alcohol consumption in male Sprague-Dawley rats were studied. Intakes of a 10% alcohol solution in milliliters per 100 gm. of body weight were on recorded weekly. No difference was found in the alcohol intake of rats given either a morphine-supplemented diet (0.01% of the diet) or daily injection of morphine sulfate (2 mg./kg.); however, there was a significant lower intake of water (p < .05) in those rats fed the diet with morphine. Total fluid consumption of both groups given the morphine-supplemented diet and daily injection of morphine sulfate was decreased (p < .05). It was found that morphine did not affect the percentage of total fluid consumed as alcohol or the weight gain.

Two levels of codeine phosphate (10 mg./kg. and 20 mg./kg.) were injected subcutaneously into rats twice a day. It was observed that the codeine significantly decreased alcohol intake in rats …