Biochemistry, Biophysics, and Structural Biology Commons^™

Laminin Receptors For Neurite Formation, H. K. Kleinman, Roy C. Ogle, F. B. Cannon, C. D. Little, T. M. Sweeney, L. Luckenbill-Edds

School of Medical Diagnostics & Translational Sciences Publications

Laminin, a basement membrane glycoprotein promotes both cell attachment and neurite outgrowth. Separate domains on laminin elicit these responses, suggesting that distinct receptors occur on the surface of cells. NG108-15 neuroblastoma-glioma cells rapidly extend long processes in the presence of laminin. We report here that 125I-labeled laminin specifically binds to these cells and to three membrane proteins of 67, 110, and 180 kDa. These proteins were isolated by affinity chromatography on laminin-Sepharose. The 67-kDa protein reacted with antibody to the previously characterized receptor for cell attachment to laminin. Antibodies to the 110-kDa and 180-kDa bands demonstrated that the 110-kDa protein …

Improved Selective Medium For The Isolation Of Treponema Hyodysenteriae, Robert A. Kunkle, J. M. Kinyon

School of Veterinary and Biomedical Sciences: Faculty Publications

An agar medium with improved selection for Treponema hyodysenteriae was developed. Cultures of T. hyodysenteriae and T. innocens, feces from 11 clinically normal pigs, and colonic contents from 6 pigs with gross lesions consistent with swine dysentery were diluted in phosphate-buffered saline and plated on Trypticase soy agar (BBL Microbiology Systems, Cockeysville, Md.) with 5% citrated bovine blood (TSA), TSA with 400 ,ug of spectinomycin per ml (TSA-S400), TSA-S400 with 25 ,ug each of colistin and vancomycin per ml, and TSA with 5% pig feces extract and five antimicrobial agents (spiramycin, rifampin, vancomycin, colistin, and spectinomycin) (BJ). Viable numbers …

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 …

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 …

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 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 …

An Immunochemical Analysis Of Alcoholic Hyaline And Its Relationship To Keratin Intermediate Filaments, David L. Drexler

Loma Linda University Electronic Theses, Dissertations & Projects

Alcoholic Hyaline (AH) and keratin intermediate filaments possess ultrastructural, biochemical and immunochemical similarities. This study compares human AH, human stratum corneum keratin (HSCK) and normal human liver cytokeratin (HCK) by quantitative immunochemical means, using polyclonal as well as monoclonal antibodies. Enzyme-linked immunosorbent assays (ELISAs) were employed, as well as "immunoblotting" and immunocytochemical techniques. HCK was isolated using Triton X-100 and "high-salt” buffers and exhibited seven major polypeptide bands on SDS-PAGE (41-58 kd). Antisera were made toward HCK and the two major HCK bands. Other immunochemicals included antisera toward AH, the three major AH bands, and HSCK, as well as 3 …

Interaction Of Putative Estrogens And The Estrogen Receptor System In Leydig Cells In The Balb/C Mouse Testis Resulting In The Initiation Of Dna Synthesis, R. Lloyd Juriansz

Loma Linda University Electronic Theses, Dissertations & Projects

Continuous administration of estrogens for 7-9 months, both steroidal and nonsteroidal, to male BALB/c mice, leads to the formation of testicular Leydig cell tumors. Three days following the subcutaneous implantation of a pellet of estrogen in cholesterol, there is a peak in the incorporation of ³H-tymidine into the DNA of the interstitial cells. These effects are hypothesized to be mediated by the estrogen receptor system in the Leydig cell. Common experimental techniques for the measurement of hormone binding, such as dextran coated charcoal treatment, proved to be impossible to employ in this system, therefore a procedure was developed using …

The Effects Of Luteinizing Hormone And Adenosine 3',5'-Cyclic Monophosphate On Phospholipid Metabolism By Luteal Mitochondria, Jim John Sadighian

Chemistry & Biochemistry Theses & Dissertations

Luteinizing hormone (LH) increases intracellular concentrations of adenosine 3', 5 '-cyclic monophosphate and the phosphoinositides, phosphatidylinositol (PI), PI 4'-phosphate (PIP) and PI 4' .5 1 - bispbosphate (PIP₂). It is believed that cAMP and the phosphoinositides act concertedly to regulate mitochondrial conversion of cholesterol to pregnenolone. This study examined the effects of LH and N⁶ ,O² -dibutyryl cAMP (dbcAMP) on phospholipids metabolism by luteal mitochondria and the influence of dbcAMP and the phosphoinosi tides on mitochondrial steroid production. Mitochondria were isolated from unincubated and incubated luteal tissue by differential centrifugation. Phospholipids were extracted from the mitochondria …

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 …

Annual Ryegrass Toxicity Research Update, A G P Brown, P. Vogal

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

Annual ryegrass toxicity (ARGT) is a complex disease that kills sheep and cattle grazing pastures which contain infected ryegrass. The disease results when a nematode (Anguina agrostis) and a batcerium (Corynebacterium sp.) invade annual ryegrass and cause the seed heads to become toxic as the grass dries off.

The bacterium produces a complex toxin of 18 glycolipid compounds or corynetoxins which have been found to be virtually identical to the antibiotic tunicamycin.

Formation Of Influenza Virus Particles Lacking Hemagglutinin On The Viral Envelope, Asit K. Pattnaik, Donald J. Brown, Debi P. Nayak

School of Veterinary and Biomedical Sciences: Faculty Publications

We investigated the intraceUular block in the transport of hemagglutinin (HA) and the role of HA in virus particle formation by using temperature-sensitive (Is) mutants (1s134 and 1s61S) of inOuenza virus AlWSN/33. We found that at the nonpermissive temperature (39.5°C), the exit of ts HA from the rough endoplasmic reticulum to the Golgi complex was blocked and that no additional block was apparent in either the exit from the Golgi complex or post-Golgi complex transport. When MDBK ceUs were infected with these mutant viruses, they produced noninfectious virus particles at 39.5°C. The efficiency of particle formation at 39.5°C was essentiaUy …

The Relationship Of Vitamins B-6 And B-12 To Plasma Homocysteine Levels In Men At Low- And High-Risk For Coronary Heart Disease, Marlene E. Swift

Loma Linda University Electronic Theses, Dissertations & Projects

There is evidence that homocysteine may be a factor in increasing the risk of coronary heart disease (CHD). To explore this relationship further, we assessed the interrelation of dietary vitamins B-6, B-12 and folate, and plasma pyridoxal-5'-phosphate and vitamin B-12 with plasma free and protein-bound homocysteine levels. Fasting blood and three-day dietary records were obtained from nine men low-risk (LR) and five at high-risk (HR) for CHD. The HR mean systolic blood pressure, total cholesterol (TC), and TC/cholesterol ratios were significantly higher than LR levels. Groups were comparable by age, weight, height, skinfolds, exercise, and smoking history. No significant difference …

Autoregulation Of Collagenase Production By A Protein Synthesized And Secreted By Synovial Fibroblasts: Cellular Mechanism For Control Of Collagen Degradation, Constance Brinckerhoff, Mary Benoit, William Culp

Dartmouth Scholarship

No abstract provided.

Lectins, El-Nasir Lalani, M Wells

Department of Pathology and Laboratory Medicine

No abstract provided.

Methemoglobin Reduction In The Presence Of Various Concentrations Of Oxyhemoglobin, Ali Mansouri

Journal of the Arkansas Academy of Science

Methemoglobin reduction in human red blood cell is governed by many factors, one of which is the concentration of oxyhemoglobin. Experiments were conducted to determine the inhibition of methemoglobin reduction by oxyhemoglobin under concentrations approaching those in the red cell. Hemoglobin, methemoglobin reductase and cytochrome b₅ were prepared by routine protein purification methods. Methemoglobin reduction was carried out at 37° C in spectrophotometer. It is shown that methemoglobin concentration variation (at least when it is above 1 mM) does not change the reduction rate. Whereas, the ratio of methemoglobin to oxyhemoglobin determines the rate of methemoglobin reduction when all other …

Negative Cooperatively In The Binding Of 3-Aminopyridine Adenine Dinucleotide To L-Α-Glycero-3-Phosphate Dehydrogenase, David Wheeler

Chemistry & Biochemistry Theses & Dissertations

Glycerol-3-phosphate dehydrogenase catalyzes the reversible reaction:

NAD⁺ + G-3-P ⇌ NADH + DHAP + H⁺

The enzyme is found in organisms from E. coli to man. The primary function of the enzyme varies from organism to organism. Depending on the metabolism of the organism, the enzyme may function primarily to generate G-3-P for lipid (in organisms with a highly aerobic biosynthesis metabolism), or it may function primarily to produce NAD⁺ to support glycolysis. Complex multicellular organisms possess tissue specific isozymes of GPDH whose kinetic parameters reflect the relative importance of G-3-P and NAD⁺ production in various …

Extrathyroidal Human Biochemistry Of Monoiodotyrosine, Stanley Andrew Tan

Loma Linda University Electronic Theses, Dissertations & Projects

The existence of extrathyroidal monoiodotyrosine (MIT) has been debated because of uncertain results from measuring serum MIT. The reason is that the isotopic and chromatographic assays used have not been sensitive enough. This investigation undertook to establish the existence of MIT in serum utilizing a radioimmunoassay (RIA), to study the physiology of serum MIT, and to discover the pharmacologic effects of exogenous MIT.

MIT-binding antibody was raised in rabbits by immunization with an MIT-porcine thyroglobulin conjugate. The antiserum bound 40% of the labeled MIT at a final dilution of 1:1000. Incubation of 100 µ1 antiserum with 50 pg [^{125 …}

The Effects Of Tri-O-Cresyl Phosphate On 2',3'-Cyclicnucleotide-3'-Phosphohydrolase Activity In Neural And Non-Neural Tissue Of The Rat, Mark K. Schlager

Chemistry & Biochemistry Theses & Dissertations

A total of 44 Long-Evans rats received 5.7 g/kg tri-O-cresyl phosphate (TOCP) orally every 3 days for 6 days. A second group of 13 rats received 4.6 g/kg orally every 3 days for 6 days. Animals were administered atropine sulfate (10 mg/kg) intraperitoneally every 6 hours or as needed to alleviate cholinergic symptoms. Ten control animals were given corn oil orally. Eight rats (18%) survived the 5.7 g/kg dose regimen and 5 rats (38%) survived the 4.6 g/kg dose regimen. Surviving rats were killed at 30-33 days post-dose. Signs of delayed neuropathy included ataxia and motor impairment.

The CNP activity …

Two-Dimensional Electrophoretic Studies Of Normal Spematozoal Proteins, Ying Lin

Chemistry & Biochemistry Theses & Dissertations

Protein profiles of human spermatozoa have been mapped by using high resolution two-dimensional electrophoresis which combines isoelectric focusing as the first dimension and polyacrylamide gradient gels electrophoresis in SDS as the second dimension. The profiles of normal spermatozoa! extraction, acrosomal extraction and seminal plasma showed high resolution in its separation capability. No qualitative or quantitative differences were observed by using Coomassie Brilliant Blue R-250 staining in a much higher concentration of proteins. However, coomassie blue lacks the sensitivity to detect many minor components. Therefore, a silver staining procedure was used, which resulted in greatly increased sensitivity.

Samples from normal fertile …

Investigation Of The Coenzyme Binding Site Of Lactate Dehydrogenase From Chicken Muscle, Jia-Hwei Lin

Chemistry & Biochemistry Theses & Dissertations

Investigation of the coenzyme binding site of lactate dehydrogenase was approached by using coenzyme analogs. Inhibition studies of this enzyme using n-alkylphosphates as coenzyme analogs were performed with a fluorometric method. The results showed that all of the n-alkylphosphates were competitive inhibitors with respect to NAD⁺.

The n-alkyl-ADPs were synthesized from AMP and dicycloheaylammonium salts of n-alkylphosphates. The synthetic compounds were subjected to NMR spectroscopic studies, thin layer chromatography and determination of the ratio of adenosine moiety to phosphate groups. The results indicated that the synthetic compounds were n-alkyl-ADPs.

These synthetic compounds were used as coenzyme analogs in …

A Method For The Comparison Of Hdl Containing Apoprotein E And Hdl Cholesterol, Steven A. Knizner

Chemistry & Biochemistry Theses & Dissertations

Human serum high density lipoproteins (HDL) were isolated by preparative salt density gradient ultracentrifugation and analyzed further by heparin-sepharose affinity chromatography. Separation of the major classes of serum lipoproteins was achieved after a single ultracentrifugation on a salt density gradient at 272,000 x g for 36 hours in a swinging bucket rotor. After isolation, the total HDL was subdivided into HDL containing apoprotein E and HDL without this apoprotein by heparin-sepharose affinity chromatography. Relative amounts of these subclasses were then calculated.

HDL-cholesterol was measured after precipitation of LDL and VLDL by phosphotungstic acid and magnesium. The statistical relationship between HDL …

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 …

The Synthesis Of Various Substituted 3-Amino-7-Hydroxy-2,2-Dimethyltetralins And Their Opioid-Related Activities, David Alan Lippman

University of the Pacific Theses and Dissertations

A series of aminotetralones and aminotetralins were synthesized from the common intermediate F, 3-amino-2,2-dimethyl-7-methoxy- 1-tetralone. The final compounds derived from F were simple substituted and/or reduced analogues. The products would allow a progressive structure activity relationship to be drawn based on pharmacological testing.

The common intermediate F was synthesized utilizing a six step procedure starting with p-methoxyphenylacetic acid. The overall yield from the precursor to the F:HC1 was 25%. Compound F was either O-demethylated to form 3-amino-2,2-dimethyl-7-hydroxy-l-tetralone (I) or was dimethylated on the amine and subsequently O-demethylated to yield the 3-dimethylamino-7-hydroxy-2,2-dimethyl-l-tetralone (J). The last major modification was the reduction of …

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 …

Identification Of Four Complementary Rna Species In Akabane Virus-Infected Cells, Asit K. Pattnaik, G. Abraham

School of Veterinary and Biomedical Sciences: Faculty Publications

The analysis of RNA extracted from purified Akabane virus demonstrated the presence of three size classes of single-stranded RNAs with sedimentation coefficients of 31S (large, L), 26S (medium, M), and 13S (small, S). Molecular weights of these RNA species were estimated to be 2.15 x 10⁶,1.5 X 10⁶, and 0.48 x 10⁶ for the L, M, and S RNAs, respectively. Hybridization analysis involving viral genomic RNA and RNA from virus-infected cells resulted in the identification of four virus-specific cRNA species in infected cells. These cRNAs were found to be nonpolyadenylated by their inability to bind …

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 …