Biochemistry Commons^™

Methods For Biological, Chemical, And Physical Analyses In Reservoirs, Penelope E. Kellar, Sherell A. Paulson, Larry J. Paulson

Publications (WR)

This manual contains detailed descriptions of biological, chemical and physical sampling and analytical procedures used at the Lake Mead Limnological Research Center. The manual was prepared in response to requests made by numerous individuals involved in the Las Vegas Valley Water Quality Program. This manual represents what we consider to be a reasonably complete compilation of methods useful in studying both general and specific limnological questions. While each section is sufficiently detailed to be of use to investigators with little experience in limnological methodology, the methods can be used on a wide variety of lakes and reservoirs, with relatively little …

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 …

Functions Of Deadenylation Factors In Mrna Decay And Mrna Processing Body Formation, Dinghai Zheng

Dissertations & Theses (Open Access)

Most newly synthesized messenger RNAs possess a 5’ cap and a 3’ poly(A) tail. The process of poly(A) tail shortening, also termed deadenylation, is important for post-transcriptional gene regulation, because deadenylation not only leads to mRNA translational inhibition but also is the first step of major mRNA degradation. Translationally inhibited mRNAs can be stored and/or degraded in dynamic cytoplasmic foci termed mRNA processing bodies, or P bodies, which are conserved in eukaryotes.

To shed new light on the mechanisms of P body formation and P body functions, I focused on the link between deadenylation factors and P bodies. I found …

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 …

The Analysis Of S-Adenosylmethionine And S-Adenosylhomocysteine In Normal And Neoplastic Cells, Kathryn Elizabeth Godburn

Chemistry & Biochemistry Theses & Dissertations

Altered patterns of methylation have been shown in both ribosomal and transfer RNA isolated from cancer cells. In contrast to the elevated activity of the methyltransferases responsible for this modification, hypomethylation appears as a general characteristic. The endogenous levels of S-adenosylmethionine and S-adenosylhomocysteine are important in relationship to their role as substrate and product of the transmethylation reaction. The levels of S-adenosylmethionine and S-adenosylhomocysteine were determined using phosphocellulose cation exchange or high pressure liquid chromatography. High voltage paper electrophoresis was used in a modified procedure to directly resolve (³⁵s) labeled S-adenosylmethionine and S-adenosylhomocysteine in small quantities of cell …

Nutrient Interactions Among Reservoirs On The Colorado River, Larry J. Paulson, John R. Baker

Publications (WR)

Interactions among physical, chemical and biological processes in reservoirs can significantly alter the characteristics of the discharge (Neel 1963, Wright 1967, Hannan 1979) that, in turn, can influence the ecology of the river downstream .(Ward and Stanford 1979). Investigations of the Colorado River, system reveal that reservoir-induced changes in the river can also affect downstream reservoirs. The formation of Lake Powell, in 1963 was accompanied by reductions in suspended sediment and nutrient loading and changes in the seasonal temperature and discharge cycles of the Colorado River. In this paper, we evaluate how these changes have influenced the nutrient and trophic …

The Effect Of Glyoxylate On Photosynthesis And Photorespiration By Isolated Soybean Mesophyll Cells, David J. Oliver

David J. Oliver

Incubating isolated soybean leaf mesophyll cells with glyoxylate increased the rates of CO2 fixation by as much as 150%. In order to cause this stimulation, the glyoxylate must be presented to the cells before the NaHCO3. Significant stimulation was observed 15 seconds after beginning the glyoxylate treatment. The glyoxylate-dependent stimulation was increased by high O2 concentrations and decreased by high CO2 concentrations. Glyoxylate treatment resulted in a 71% inhibition in the rate of CO2 incorporation into glycolate and glycine. Glyoxylate may be stimulating net photosynthesis solely by decreasing photorespiration or it may be increasing the amount of CO2 fixed by …

Studies Of The Reorientational Relaxation Of Pyridine In Water By Depolarized Rayleigh Light Scattering, C. H. Wang, Scott L. Whittenburg, P. A. Lund, D. H. Christensen

Chemistry and Biochemistry Faculty Publications

The depolarized Rayleigh spectra of aqueous solutions of pyridine have been studied using a high‐finesse Fabry–Perot interferometer as a function of temperature and concentration. The Rayleigh relaxation times are found to have a complex concentration and viscosity dependence. The classical Stokes–Einstein–Debye equation for molecular reorientation breaks down in this system. The Rayleigh relaxation time of pyridine molecules is not determined by the macroscopic shear viscosity of the solution. The specific interaction due to the formation of hydrogen bonds between pyridine and water molecules plays a very important role in affecting the relaxation time. At a fixed temperature the plot of …

Structural Studies Of Preimplantation Rabbit Embryos Using Ftir Atr, Amy R. Moler-Booth

Chemistry & Biochemistry Theses & Dissertations

FTIR was used in conjunction with a surface selective technique, known as attenuated total reflectance (ATR), to study the infrared spectra of preimplantation rabbit embryos. The rabbit embryos ranged in aged from 3 to 6 days. Since an embryo is mostly water, deuteration of the embryo was necessary to eliminate the intense water background present in the mid-infrared region. A set of spectral standards were tested, in order to identify the bands seen in embryo spectra. By analyzing and comparing the spectra from 3, 4, 5 and 6 day embryos, differences could be noted. In particular, bands defined as Amide …

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 …

Adaptative Alteration In Phospholipid Composition Of Plasma Membranes From A Somatic Cell Mutant Defective In The Regulation Of Cholesterol Biosynthesis, Michael Sinensky

Faculty Publications, Biological Sciences

A somatic cell mutant (CR1) of a Chinese hamster ovary cell (CHO-K1) which has previously been shown to be defective in the regulation of cholesterol biosynthesis accumulates more cholesterol than the parental cell line in plasma membranes. Although such an increase in membrane cholesterol should lead to an increase in the order parameter of these membranes, as measured with an electron spin resonance spin probe, the order parameters of mutant and wild-type plasma membranes are identical--apparently because of an adaptive alteration in membrane phospholipid composition. The phospholipid compositions of mutant and wild-type cell plasma membranes are compared and the mutant …

Oxidative Metabolism Of 4-Aminobutyrate By Rat Brain Mitochondria: Inhibition By Branched-Chain Fatty Acid / J. Cunningham, D. D. Clarke, And W.J. Nicklas, J. Cunningham, Donald Dudley Clarke Phd, William J. Nicklas

Chemistry Faculty Publications

The oxidation of 4-aminobutyric acid (GABA) by nonsynaptosomal mitochondria isolated from rat forebrain and the inhibition of this metabolism by the branched-chain fatty acids 2-methyl-2-ethyl caproate (MEC) and 2,2- dimethyl valerate (DMV) were studied. The rate of GABA oxidation, as measured by 02 uptake, was determined in medium containing either 5 or 100 mM-[K+]. The apparent Km for GABA was 1.16 ± 0.19 mM and the Vmax in state 3 was 23.8 + 5.5 ng-atoms O2 ·min-1.mg protein-1 in 5 mM-[K+]. In a medium with 100 mM-[K+] the apparent K111 was 1.11 + 0.17 mM and V max was 47.4 …

Aspects Of Larval Ecology Of Squilla Empusa (Crustacea, Stomatopoda) In Chesapeake Bay, Steven G. Morgan

OES Faculty Publications

Larvae of Squilla empusa were collected from the plankton and were laboratory-reared in 16 combinations of temperature and salinity to determine their tolerances. Larvae survived longer and molted more frequently when reared at 25%, and 20° or 25° C, which corresponds to the natural conditions of Chesapeake Bay when the larvae were collected.

A 2 year planktonic survey conducted in the lower region of the bay by the Virginia Institute of Marine Sciences was compared with a survey made at the bay mouth in 1976. The seasonal occurrence of Squilla empusa larvae extended from the last week of July until …

Anaerobic Metabolism Of The Ribbed Mussel, Geukensia Demissa, Ming-Shan Ho

Dissertations, Theses, and Masters Projects

The anaerobic metabolism of the intertidal ribbed mussel, Geukensia demissa (= Modiolus demissus), is reported in this dissertation. This mollusc was selected because anaerobic metabolism is believed to be of vital significance to its survival during the air exposure period of the tidal cycle.;Ribbed mussels, Geukensia demissa, were maintained in either aerated or hypoxic water for up to four days. Major metabolites and polysaccharide of each mussel were then analyzed and compared. Succinate, propionate and alanine were found to accumulate hypoxically, but not malate, (alpha)-keto-glutarate and pyruvate. The polysaccharide content of each mussel was quite different and did not show …

Multiple Methods Of Analysis Of 2'-O-Methylation In 5.8s Rrna, Sharon H. Ryan

Chemistry & Biochemistry Theses & Dissertations

5.8S rRNA is a low molecular weight ribosomal RNA which is found hydrogen bonded to 28S rRNA in the eucaryotic cell. There are two nucleotides which have 2'-0-methylations; a guanine residue at position 77 is fully methylated and a uridine residue at position 14 which is partially methylated. This partial 2'-0-methylation of the uridine residue has been found to vary with the tissue source, with the highest level in normal tissue and the lowest level in neoplastic tissue. In order to study the significance of this site-specific methylation, a reliable and convenient method of analysis was needed.

Early studies on …

Adaptative Alteration In Phospholipid Composition Of Plasma Membranes From A Somatic Cell Mutant Defective In The Regulation Of Cholesterol Biosynthesis, Michael Sinensky

Michael Sinensky

A somatic cell mutant (CR1) of a Chinese hamster ovary cell (CHO-K1) which has previously been shown to be defective in the regulation of cholesterol biosynthesis accumulates more cholesterol than the parental cell line in plasma membranes. Although such an increase in membrane cholesterol should lead to an increase in the order parameter of these membranes, as measured with an electron spin resonance spin probe, the order parameters of mutant and wild-type plasma membranes are identical--apparently because of an adaptive alteration in membrane phospholipid composition. The phospholipid compositions of mutant and wild-type cell plasma membranes are compared and the mutant …

The Association Of 5.8 S With 28s Ribosomal Rna, Nandita Banerjee

Chemistry & Biochemistry Theses & Dissertations

5.8S rRNA is a low molecular weight ribosomal RNA which is noncovalently bonded to the larger ribosomal subunit 28S rRNA; through its 3' end and through its 5' end. This interaction is an integral part of the ribosome, and plays an important role in the ribosome structure and function.

There is a high degree of homology between the 5.8S rRNA primary structures of rat, turtle and chicken. The base sequence of rat 5.8S rRNA differs only in one position from that of turtle and in three positions fr.om that of cl1.icken. Tl1ere is a single purine substitution at the 5' …