Life Sciences Commons^™

Spermatogenesis In The Mosquito Eretmapodites Quinquevittatus, Robert Daniel Hunter Jr.

Legacy ETDs

Various stages of spermatogenesis in pupal testes of the mosquito Eretmapodites quinquevittatus (Theobald) were determined by 2% 1acto-aceto-orcein and silver nitrate staining techniques. Temporal relationships of stages were demonstrated and behavior and appearance of meiotic chromosomes were described.

Pupal testes were most active meiotically for the first 12-14 hours after onset of pupation. The prepachytene stage of leptotene and a nucleolus were observed with the use of silver nitrate staining. Experimental observations demonstrated the first spermatids and visible mature sperm at 6-8 and 18 hours, respectively, after onset on pupation. E. quinquevittatus mean sperm size was reported (head = 50 …

A High Molecular Weight Sulfhydryl Dependent Proteinase From Bovine Spleen, Thomas J. Farrell

Master's Theses

No abstract provided.

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

Chemistry Faculty Publications

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

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

Chemistry Faculty Publications

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

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

Chemistry Faculty Publications

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

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

Department of Biochemistry: Faculty Publications

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

Short-Term Feedback Regulation Of Camp By Accelerated Degradation In Rat Tissues, Tom W. Gettys, Peter F. Blackmore, J. Bruce Redmon, Stephen J. Beebe, Jackie D. Corbin

Bioelectrics Publications

A recent study showed that cAMP analogs lowered cAMP levels in rat hepatocytes. The present work demonstrates that cAMP analogs also lowered cAMP in a rapid, concentration-dependent manner in heart and fat cells. In order to determine if the cAMP-dependent protein kinase mediated this effect, techniques were developed to assay the protein kinase activity ratio in hepatocytes treated with cAMP analogs. The activation of protein kinase and phosphorylase in hepatocytes by 8-pClΦS-cAMP (where 8-pClΦS- indicates 8-parachlorothiophenyl-) was concentration-dependent and occurred in parallel to proportionate decreases in cAMP. More than 20% of the cAMP binding sites on the protein kinase were …

Photoreactivation Of Lethal Damage Induced In Hamster X Xenopus Hybrid Cells And Their Parentals By Uv Light, David Bohlender, Stuart Williams, Emir Cruz, H. Gaston Griggs

Journal of the Arkansas Academy of Science

A85 Xenopus cells that exhibited a high level of photoreactivation (PR) and V79B2 hamster cells that exhibited little PR were fused to produce the V79B2 x A85 cell line — a hybrid line which possessed a relatively stable karyotype, with most cells containing the entire V79B2 and A85 genomes. UV and UV plus PR fluence-survival relations were then determined and compared for the hybrid and parental lines in a first attempt to elucidate interactions of the parental PR mechanisms in the hybrid. It was anticipated that the A85 genome in the hybrid would produce PR enzyme in sufficient concentration and …

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

Journal of the Arkansas Academy of Science

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

Spermatogenesis And Histology Of The Testes Of The Caecilian, Chthonerpeton Indistinctum, Rafael O. De Sá, Nibia Berois

Biology Faculty Publications

Macroscopically, the testes of Chthonerpeton indistinctum consist of a series of oval lobes. The number and size of lobes is variable in Chthonerpeton indistinctum. Histologically, the testes are divided into structural units, locules separated by septa of connective tissue. Inside of each locule spermatogenesis takes place in cysts of germinal cells that divide synchronically. Chthonerpeton indistinctum is divided chronologically into four stages of development based on the shape of the nuclei of the spermatids and their arrangement in the cyst: (1) round, (2) kidney shaped, (3) attenuate, and (4) bouquet arrangement.

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 …

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 …

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

Loma Linda University Electronic Theses, Dissertations & Projects

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

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 …

Ferroxidase Activity Of Mushroom Tyrosinase., Rodney Boyek, David Mascotti, Barbara Schori

David P. Mascotti

Mushroom tyrosinase catalyses the oxidation of Fe(II) to Fe(III). Both the newly-discovered ferroxidase and the well-characterized diphenol oxidase activities of tyrosinase exhibit inhibition by cyanide and both activities co-purify during two preparation steps. The characteristics of tyrosinase-catalysed Fe(II) oxidation are compared with those of other ferroxidases.

Auger Analysis Of Si–H Bonding And Hydrogen Concentration In Hydrogenated Amorphous Silicon, Nancy Burnham, Aj Nelson, Ab Schwartzlander, Se Asher, Ll Kazmerski

Nancy A. Burnham

Auger electron spectroscopy line‐shape analysis of the Si‐L 2 3 V V peak has been performed on hydrogenated amorphous silicon (a‐Si:H). Both a‐Si:H produced by hydrogen implantation of siliconsingle crystals (for analytical standards) and thin films (fabricated for solar cell applications) were examined in these studies. Hydrogen concentrations were confirmed by secondary ion mass spectrometry, and samples having hydrogen content over the range 101 6–102 2 cm− 3 were evaluated. Correlations between the area under the deconvoluted L 2 3 V V transition peak and the known hydrogen content have resulted in a semiquantitative method of determining hydrogen concentration using …

Yeast Mannans: Use As A Transport Vehicle For Peptides, Jacqueline Cesario Lee

Masters Theses

Yeast mannans are phosphorylated glycopeptides found in various species of yeast. Their similarity to the mannose-phosphate signal for intracellular lysosomal uptake led mannans to be considered as a possible vehicle for increasing cellular affinity for peptides. Insulin was used as a model peptide and was conjugated to mannans by two coupling methods. The conjugated mannan-insulins were compared to insulin on the basis of cellular binding and ability to induce typical insulin biological effects.

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 …

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 …

Purification And Characterization Of A Soluble And A Particulate Glutamate Dehydrogenase From Rat Brain / Arlene D. Colon, Andreas Plaitakis, Antonis Perakis, Soll Berl, And Donald D. Clarke Department Of Neurology, Mount Sinai School Of Medicine, New York, And Department Of Chemistry, Fordham University, Bronx, New York, U.S.A., Arlene D. Colon, Andreas Plaitakis, Antonis Perakis, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Glutamate dehydrogenase (GDH) activity was determined in high-speed fractions (1 00,000 g for 60 min) obtained from whole rat brain homogenates after removal of a low-speed pellet (480 g for 10 min). Approximately 60% of the high-speed GDH activity was particulate (associated with membrane) and the remaining was soluble (probably of mitochondrial matrix origin). Most of the particulate GDH activity resisted extraction by several commonly used detergents, high concentration of salt, and sonication; however, it was largely extractable with the cationic detergent cetyltrimethylammonium bromide (CTAB) in hypotonic buffer solution. The two GDH activities were purified using a combination of hydrophobic …

The Subcellular Localization Of Glutamate Dehydrogenase (Gdh): Is Gdh A Marker For Mitochondria In Brain? / James C. K. Lai, Kwan-Fu Rex Sheu, Young Tai Kim, Donald D. Clarke, And John P. Blass Department Of Neurology, Cornell University Medical College And Altschul Laboratory For Dementia Research Burke Rehabilitation Center White Plains, Ny 10605 And Department Of Medicine Cornell University Medical College New York, Ny 10021, James C. K. Lai, Kwan-Fu Rex Sheu, Young Tai Kim, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Glutamate dehydrogenase (GDH, EC 1.4.1.2) has long been used as a marker for mitochondria in brain and other tissues, despite reports indicating that GDH is also present in nuclei of liver and dorsal root ganglia. To examine whether GDH can be used as a marker to differentiate between mitochondria and nuclei in the brain, we have measured GDH by enzymatic activity and on immunoblots in rat brain mitochondria and nuclei which were highly enriched by density-gradient centrifugation methods. The activity of GDH was enriched in the nuclear fraction as well as in the mitochondrial fraction, while the activities of other …

Quantitation And Identification Of Organic N-Chloramines Formed In Stomach Fluid On Ingestion Of Aqueous Hypochlorite, Frank E. Scully Jr., Katherine Mazina, Daniel E. Sonenshine, Frederick Kopfler

Biological Sciences Faculty Publications

The chemical reactions that hypochlorite undergoes in the body when chlorinated water is ingested have received very little attention. Because amino nitrogen compounds are important components of the average diet, the reactions of hypochlorite with amino compounds in the stomach were investigated. Stomach fluid was recovered from Sprague-Dawley rats that had been fasted for 48 hr and administered 4 mL deionized water. The chlorine demand of the stomach fluid was determined. An average volume-independent demand of 2.7 mg chlorine was measured. At doses below 40 mg/L chlorine reducing reactions appeared to account for reduction of all oxidizing species within 15 …

Investigation Of Nickle As A Matrix Modifier In The Atomic Absorption Analysis Of Tin, Charles William Saunders

Chemistry & Biochemistry Theses & Dissertations

Nickel has been widely used as a matrix modifier in the flameless atomic absorption analysis of arsenic and selenium. Its utility as a modifier for tin has been investigated using Polarized Zeeman effect Graphite Furnace Atomic Absorption Spectroscopy. Peak area, peak shape, and appearance times were compared in modified and nonmodified standards using the system computer's internal integration and a detector/readout system with 18 msec resolution.

The addition of nickel at 600 mg/L increased the tin absorbance signal five-fold. The improved signal to noise ratio resulted in a two-fold increase in the sensitivity and a lower limit of detection. The …

Localization Of A Camp-Dependent Protein Kinase In Ovarian Mitochondria, William Jameson Roberts

Dissertations, Theses, and Masters Projects

No abstract provided.

In Vitro Phosphorylation Of Maize Leaf Phosphoenolpyruvate Carboxylase, Raymond J. A. Budde, Raymond Chollet

Department of Biochemistry: Faculty Publications

Autoradiography of total soluble maize (Zea mays) leaf proteins incubated with ³²P-labeled adenylates and separated by denaturing electrophoresis revealed that many polypeptides were phosphorylated in vitro by endogenous protein kinase(s). The most intense band was at 94 to 100 kilodaltons and was observed when using either [γ -32P]ATP or [β-³²P]ADP as the phosphate donor. This band was comprised of the subunits of both pyruvate, Pi dikinase (PPDK) and phosphoenolpyruvate carboxylase (PEPCase). PPDK activity was previously shown to be dark/light-regulated via a novel ADP-dependent phosphorylation/Pi-dependent dephosphorylation of a threonyl residue. The identity of the acidstable 94 …

Isolation Of Nuclei From Physarum Flavicomum: Demonstration Of Nuclear Cyclic Acid Amp Phosphodiesterase, Judith A. Bean, Maurice G. Kleve, Thomas J. Lynch

Journal of the Arkansas Academy of Science

Cyclic AMP phosphodiesterase activity in the nucleus of the myxomycete Physarum flavicomum was demonstrated by cytochemical staining utilizing electron microscopy and by enzymatic assays with tritiated cyclic AMP as the substrate. Cytochemical staining showed Physarum's plasmodial phosphodiesterase activity to be located in the nucleus, along the plasma membrane, in vesicles, and free in the cytoplasm. Nuclear phosphodiesterase, which may be cell cycle dependent, was primarily located in the nucleolus. Nuclei from three to five day old microplasmodial cultures were isolated by the method of Henney and Yee. Whole cells were collected through centrifugation and washed. Pellets were homogenized in a …

Concanavalin A-Nonbinding Enzymes Of Crotalus Scutulatus Scutulatus Venom, C. K. Childs, Michael W. Hinson, Dewey H. Sifford, Bob D. Johnson

Journal of the Arkansas Academy of Science

Crotalus scutulatus scutulatus crude venom was separated into two fractions by Concanavalin A Sepharose 4B affinity chromatography. The Concanavalin A-nonbinding fraction (F-l) exhibited phosphomonoesterase (orthophosphoric monoester phosphohydrolase EC 3.1 .3.2), phosphodiesterase, 5 '-nucleotidase (5 '-ribonucleotide phosphohydrolase EC 3.1.3.5), phospholipase A (phosphatidate 2-acylhydrolase EC 3.1.1.4), hyaluronidase (hyaluronate glycanohydrolase EC 3.2.1.d), N-benzoyl-Larginine ethyl esterase, p-toluenesulfonyl-L-arginine methyl esterase, L-amino acid oxidase (L-amino acid: O2 oxidoreductase [deaminating] EC 1.4.3.2), and caseinolytic activities. Thrombin-like and NAD nucleosidase (5 '-ribonudeotide phosphohydrolase EC 3.1.3.5) activities were not observed. DEAE Sephadex A-50 ion exchange chromatography by two stage elution of F-l yielded several fractions having proteinase activities. Proteinase …

The Effects Of Various Adenosine And Xanthine Analogues On Mammalian Sperm Motility, Daniel Joseph Cushing

Masters Theses

Caffeine, theophylline, and a number of other xanthine analogues have been shown to stimulate the motility of mammalian spermatozoa. The mechanism by which these compounds act was assumed to be cAMP phosphodiesterase inhibition. However, it has recently been shown that many of the responses elicited by alkylxanthines and their analogues in various tissues are not the result of cAMP phosphodiesterase inhibition but by antagonism of endogenous adenosine. Using the recently described transmembrane migration method this study observed the effect of a number of xanthine and adenosine analogues on mammalian sperm motility. The hypothesis to be tested was that the increase …

Optimal Conditions For Kinetic Study Of Succinate Dehydrogenase In Rat Liver, Collie B. Shaw, Tara L. Chronister, John D. Peck

Journal of the Arkansas Academy of Science

Succinate dehydrogenase (SDH) commonly is assayed as a marker enzyme for mitochondrial activity. The literature presents numerous conditions for conducting this assay due to the fact that, it has been difficult to get sufficient reduction of the acceptor dye, 2,3,5-triphenyl-2H-tetrazolium chloride (TTC). This study was undertaken to optimize the SDH-catalyzed reduction of TTC dye by evaluation of a greater range of molor ratios of TTC to succinate and by further evaluation of additives reported as beneficial. Improvement in enzyme specific activity was achieved by liver perfusion via the left cardiac ventricle with homogenizing solution. Increase in TTC from 1 to …