Biochemistry Commons^™

Analysis Of Phytoestrogens By High Performance Liquid Chromatography, Anne A. Grippo, Yan Xie, Benjamin L. Rougeau, William V. Wyatt

Journal of the Arkansas Academy of Science

Phytoestrogens are biochemicals synthesized in plants which mimic steroidal estrogen activity in mammals. Analysis of these compounds in the legumes which produce them and in body fluids is important to the study of their physiological effects. High pressure liquid chromatography (HPLC) has been found to be an efficient and sensitive method of identification and quantitation of isoflavonoids, one class of phytoestrogen. Here we report the separation of three isoflavonoids, biochanin A, genistein and daidzein using an HPLC system with a Cg reverse phase column and a linear gradient mobile phase containing acetonitrile and acetic acid/water (10/90, v/v) over 60 minutes. …

Two-Photon Excitation Of Rhenium Metal-Ligand Complexes, Joseph R. Lakowicz, Felix N. Castellano, Ignacy Gryczynski, Zygmunt Gryczynski, Jonathan D. Dattelbaum

Chemistry Faculty Publications

We describe the emission spectral properties of two rhenium metal-ligand complexes with one and two-photon excitation, Re(bpy)₂(CO)₃Cl and [Re(bpy)(CO)₃CH₃CN]⁺, where bpy is 2,2’-bipyridyl and CH₃CN is acetonitrile. Similar emission spectra and intensity decay times characteristic of the metal-to-ligand charge transfer state were observed for one- and two-photon excitation. The lifetime and quantum yield of the acetonitrile complex are approximately 14-fold higher than that of the chloride complex. Both complexes display high anisotropies near 0.33 in frozen solution with one-photon excitation. Two-photon excitation results in anisotropies about 40% larger, …

Investigating The Allosteric Behavior Of Malate Dehydrogenase From Escherichia Coli, Eman M. Ghanem

Masters Theses

Regulatory mechanisms of malate dehydrogenase from E.coli (eMDH) involving NADH as an allosteric effector were investigated. The reaction was studied in both directions: malate oxidation and oxaloacetate reduction. When malate was the variable substrate, a plot of rate against substrate concentration was sigmoidal in the presence of 0.065 mM NADH, which indicates the presence of an allosteric site for NADH on the enzyme. Binding of NADH at the allosteric site causes conformational changes in the active site and, thereby, changes the catalytic activity of the enzyme. An increase in K_m value, from 1.3 to 3.9 mM malate was observed, …

Phorbol 12-Myristate 13-Acetate Stimulates Lysophosphatidic Acid Secretion From Ovarian And Cervical Cancer Cells But Not From Breast Or Leukemia Cells, Zhongzhou Shen, Jerome Belinson, Richard E. Morton, Yan Xu

Chemistry Faculty Publications

Lysophosphatidic acid (LPA) is present in ascites from patients with ovarian cancer. It stimulates calcium release and growth of ovarian cancer cells bothin vitroandin vivo.Recently, we found that LPA levels were significantly elevated in plasma from patients with ovarian cancer and other gynecological cancers. In contrast, LPA levels were not elevated in patients with breast cancer and leukemias. In view of this, we investigated whether gynecological cancer cells could produce LPA. LPA was extracted from the supernatant of cells culturedin vitroand purified by thin layer chromatography. After hydrolysis and transmethylation, the fatty acid derivatives were analyzed by gas chromatography. We …

Principal Component Mapping Applied To Raman Microspectroscopy Of Fiber-Reinforced Polymer Composites, C. M. Stellman, K. S. Booksh, A. R. Muroski, M. P. Nelson, Michael L. Myrick

Faculty Publications

We describe the development and implementation of a spectroscopic Raman imaging system using Principal Component data analysis. Raman microspectroscopy followed by multivariate data analysis and principal component mapping has been used to investigate the spatial variations of chemical and physical properties at interfaces in glass-reinforced composites on the micrometer scale. A protocol for monitoring chemical and physical variances via multivariate statistics has been developed and a preliminary study of a "real world" glass-reinforced composite is presented. It is demonstrated that multivariate Raman imaging is more precisely interpretable than traditional univariate methods and that by appropriately choosing distinct spectral regions (such …

Dynamics Of Biomolecules: Assignment Of Local Motions By Fluorescence Anisotropy Decay, Carl N. Bialik, Barnabas Wolf, Edward L. Rachofsky, J. B. A. Ross, William Laws

Chemistry and Biochemistry Faculty Publications

Many biological systems have multiple fluorophores that experience multiple depolarizing motions, requiring multiple lifetimes and correlation times to define the fluorescence intensity and anisotropy decays, respectively. To simplify analyses, an assumption often made is that all fluorophores experience all depolarizing motions. However, this assumption usually is invalid, because each lifetime is not necessarily associated with each correlation time. To help establish the correct associations and recover accurate kinetic parameters, a general kinetic scheme that can examine all possible associations is presented. Using synthetic data sets, the ability of the scheme to discriminate among all nine association models possible for two …

Monte Carlo Simulation Of An Ion-Dipole Mixture As A Model Of An Electrical Double Layer, Douglas Henderson, Dezso Boda, Kwong-Yu Chan

Faculty Publications

Canonical Monte Carlo simulations were performed for a nonprimitive model of an electrical double layer. The ions and the solvent molecules are modeled as charged and dipolar hard spheres, respectively, while the electrode as a hard, impenetrable wall carrying uniform surface charge. We found that the ion-dipole model gives a reasonable description of the double layer for partially charged ions with small to moderate dipole moments, or equivalently for an "effective" dielectric constant. Density, polarization and mean electrostatic potential profiles are reported. Strong layering structure, and at higher charges, charge inversion in the second layer were found. With appropriate choices …

Interagency Lake Mead And Las Vegas Wash Monitoring Program: Standard Operating Procedures Manual, Bureau Of Reclamation, City Of Henderson Water Reclamation Facility, City Of Las Vegas Water Pollution Control Facility, Clark County Sanitation District, Nevada, Southern Nevada Water Authority

Publications (WR)

A number of agencies sample Lake Mead and the Las Vegas Wash on a routine basis at several locations. In order to share and properly interpret the data, the Bureau of Reclamation, Southern Nevada Water Authority and the three Wastewater Treatment Facilities (City of Las Vegas, Clark County Sanitation District and City of Henderson) formed a committee to examine sampling and analytical protocols and to share information with the goal of maximizing the data quality. The group first met in April 1997. It was agreed that an effort should be made to discuss and compare specific sampling and analytical techniques …

Simultaneous Formation Of Functional Leading And Lagging Strand Holoenzyme Complexes On A Small, Defined Dna Substrate, Anthony J. Berdis, Stephen J. Benkovic

Chemistry Faculty Publications

The biochemical characterization of leading and lagging strand DNA synthesis by bacteriophage T4 replication proteins has been addressed utilizing a small, defined primer/template. The ATP hydrolysis activity of 44/62, the clamp loading complex responsible for holoenzyme assembly, was monitored during assembly of both the leading and lagging strand holoenzyme complex. The ATPase activity of 44/62 diminishes once a functional holoenzyme is assembled on both the leading and lagging strand. The assembly of the lagging strand holoenzyme is facilitated by several factors including biotinylated streptavidin blocks at the end of the fork strands, preassembly of the leading strand holoenzyme, and by …

Localization Of Sodium, Potassium-Adenosine Triphosphatase In The Cerebral Cortex Of Developing Rats, Maki Ujiie

Loma Linda University Electronic Theses, Dissertations & Projects

Active transport via Na⁺, K⁺-ATPase plays an important role in establishing and maintaining proper ionic gradient across all mammalian cell membranes. A strict regulation of Na⁺, and K⁺ ion levels is especially important in excitable cells such as neurons and myocytes. Inhibition of the enzyme is associated with several pathological conditions and has serious detrimental effects on cerebral cortical cells. Yet the enzyme appears to be limited in amount and distribution in newborn rats. In this study, Na_⁺, K⁺- ATPase expression and distribution in early postnatal rats were studied …

Solute-Solvent Pair Distribution Functions In Highly Asymmetric Additive Hard Sphere Mixtures, Douglas Henderson, Kwong-Yu Chan

Faculty Publications

Contact values for the solute-solvent pair distribution function in an additive hard sphere mixture, as computed from the Henderson-Chan (HC) formulas, are compared with the recent Monte Carlo (MC) data and formula of Matyushov and Ladanyi (ML) [J. Chem. Phys. 107, 5815 (1997)]. The agreement is found to be excellent. The negative finding of ML is due to a misprint in one of the HC publications. We find the HC formula to be superior to the ML formula when compared to our MC data [Mol. Phys. 91, 1137 (1997)] for the case where the ratio of diameters is large and …

Influences Of Acid Mine Drainage On Water Chemistry Of Blackwater River Watershed, Carla C. Morato-Lara

Theses, Dissertations and Capstones

Acidic mine drainage flow from active and abandoned coal mines has been a major source of water pollution in the Appalachian Mountain region of the eastern USA. Water samples were collected and analyzed from Blackwater River watershed during 2 years (September 1996 - May 1998). The water quality measurements determined that the Beaver Creek and North Fork tributaries were affected by acid mine drainage but not Yellow Creek. Beaver Creek and North Fork stations had high sulfate and manganese markers of acid mine drainage, whereas Yellow Creek was slightly acidic with low sulfate and manganese. North Fork was the primary …

Vibrational Sum Frequency Spectroscopy Of Surfactants And Phospholipids Monolayers At Liquid-Liquid Interfaces, B. L. Smiley, R. A. Walker, D. E. Gragson, T. E. Hannon, G. L. Richmond

Chemistry and Biochemistry

Work from our laboratory on vibrational sum frequency spectroscopic investigations of molecular ordering at the carbon tetrachloride-water interface is reviewed. Simple charged surfactants adsorbed at the liquid-liquid interface are seen to induce alignment of interfacial water molecules to a degree which is dependent on the induced surface potential. Saturation of water molecule alignment occurs at a surfactant surface concentration corresponding to a calculated surface potential of approximately 160 mV. In complementary studies, the relative degree of hydrocarbon chain ordering within monolayers of symmetric phosphatidylcholines of different chain lengths is inferred by the relative signal contributions of the methyl and methylene …

Probing The Intermolecular Hydrogen Bonding Of Water Molecules At The Ccl4/Water Interface In The Presence Of Charges Soluble Surfactant, D. E. Gragson, G. L. Richmond

Chemistry and Biochemistry

The molecular structure and hydrogen bonding of water molecules at the CCl₄/water interface in the presence of a charged soluble surfactant has been explored in this study using vibrational sum frequency generation. By examining OH stretching modes that are highly sensitive to the local hydrogen bonding environment it is found that water molecules at the CCl₄/water interface both in the presence and absence of a charged soluble surfactant are predominantly in a tetrahedral arrangement much like the structure ofice. Isotopic dilution studies have been employed to further characterize this icelike interfacial structure. Ablueshift ofapproximately120 cm^{-1 …}

Liquid-Liquid Immiscibility In Lipid Monolayers, John P. Hagen, Harden M. Mcconnell

Chemistry and Biochemistry

Some binary lipid mixtures form coexisting liquid phases when spread at the air/water interface. This work describes the pressure–composition phase diagrams of binary mixtures of four unsaturated phosphatidylcholines with dihydrocholesterol. These four binary mixtures have critical compositions of approximately fifty mole percent, and average critical exponents of 0.25±0.07. The data can also be approximated by a regular solution thermodynamic model, yielding parameters for the non-ideality of these mixtures.

A Fluid In Contact With A Semipermeable Surface: Second-Order Integral Equation Approach, Douglas Henderson, Pawel Bryk, Stefan Sokolowski

Faculty Publications

An integral equation approach for a binary hard-sphere mixture interacting with a planar semipermeable wall (membrane) is formulated by using the second-order nonuniform or pair Ornstein–Zernike equation as well as the usual singlet Ornstein–Zernike equation. The results of the pair theory are compared with those obtained from the singlet theory and with computer simulation data. The pair approach is more accurate than the singlet theory.

Emissions From Smoldering Combustion Of Biomass Measured By Open-Path Fourier Transform Infrared Spectroscopy, Robert J. Yokelson, Ronald A. Susott, Darold E. Ward, James J. Reardon, David W. T. Griffith

Chemistry and Biochemistry Faculty Publications

Biomass samples from a diverse range of ecosystems were burned in the Intermountain Fire Sciences Laboratory open combustion facility. Midinfrared spectra of the nascent emissions were acquired at several heights above the fires with a Fourier transform infrared spectrometer (FTIR) coupled to an open multipass cell. In this report, the results from smoldering combustion during 24 fires are presented including production of carbon dioxide, carbon monoxide, methane, ethene, ethyne, propene, formaldehyde, 2-hydroxyethanal, methanol, phenol, acetic acid, formic acid, ammonia, hydrogen cyanide, and carbonyl sulfide. These were the dominant products observed, and many have significant influence on atmospheric chemistry at the …

Comparison Of The Dichlorination Of Organic N-Chloramines By Caroate And By Bisulfite: A Kinetic Study, Said H. Lubbad

Chemistry & Biochemistry Theses & Dissertations

Bisulfite is commonly used to dechlorinate chlorinated municipal wastewater. However, several studies suggest that bisulfite is not effective in dechlorinating N-chlorinated amino components of wastewater. The purpose of this study was to investigate the effectiveness of an alternate dechlorinating agent, caroate (peroxysulfate), and to compare its rate of dechlorination of three different types of chloramines with that of bisulfite.

Four N-chlorinated amino compounds were investigated: N-chlorodiethylamine, N­ chloroalanylphenylalanine, N-chloroglycylphenylalanine, and N-[2-(N'-chloroimino)­ propanoyl]phenylalanine. These compounds were selected from different families of N­ chloramines to study the effect of structure on dechlorination by both caroate and bisulfite. At pH …

Speciation Of Selenium In Serum, Alice Tsapayi-Musapatike

Chemistry & Biochemistry Theses & Dissertations

We investigated the separation of selenoprotein P and extracellular (serum) glutathione peroxidase using a concanavalin A affinity column. We were able to show that the method does not adequately separate serum glutathione peroxidase from selenoprotein P because human glutathione peroxidase binds strongly to the column. We also found that albumin can interfere with measurement serum of glutathione peroxidase activity when hydrogen peroxide is used as the substrate. When t-butyl hydroperoxide was used as the substrate, less than 10% of the measured activity was due to the presence of albumin. We also found that the pH of the elution buffer plays …

Grand Canonical Monte Carlo And Modified Singlet Integral Equations For The Density Profile Of A Yukawa Fluid Near A Planar Wall, Douglas Henderson, Wilmer Olivares-Rivas, Leo Degreve, Jacqueline Quintana

Faculty Publications

Results for the density profile for Yukawa molecules near a hard wall and an exponential attractive wall are presented for Grand Canonical Monte Carlo (GCMC) simulations, for the singlet hypernetted chain (HNC) integral equation and for a modified version of the Lovett–Mou–Buff–Wertheim (LMBW-1) which uses the exact contact value theorem. The results of the standard singlet HNC are quite poor. If the LMBW equation is modified (but still using the bulk direct correlation function) the results at high temperature become reasonable. However, the results at low temperatures, close to the bulk coexistence curve, are only a partial improvement. The contact …

Laser-Initiated Decomposition Products Of Indocyanine Green (Icg) And Carbon Black Sensitized Biological Tissues, John M. Kokosa, Andrzej Przyjazny, Kenneth Eugene Bartels, Massoud Motamedi, Donald J. Hayes, David B. Wallace, Christopher J. Frederickson

Chemistry & Biochemistry Publications

Organic dyes have found increasing use a s sensitizers in laser surgical procedures, due to their high optical absorbances. Little is known, however, about the nature of the degradation products formed when these dyes are irradiated with a laser. Previous work in our laboratories has shown that irradiation of polymeric and biological tissues with CO2 and Nd:YAG lasers produces a host of volatile and semivolatile by-products, some of which are known to be potential carcinogens. This work focuses on the identification of the chemical by-products formed by diode laser and Nd:YAG laser irradiation of indocyanine green (ICG) and carbon black …

Binding Of Cobalt (Iii) To Nucleic Acids Via Reaction With [Co(Nh3)5(Oh2)]3+, David M. Calderone

Seton Hall University Dissertations and Theses (ETDs)

No abstract provided.

Synthesis And Characterization Of Modified Silica Gel As An Intermediate In The Generation Of Gaseous Standard Mixtures, M. Prokopowicz, K. Lewandowska, A. Skwierawska, Andrzej Przyjazny, Jan F. Biernat, J. Namiesnik

Chemistry & Biochemistry Publications

A possibility of extending analytical applications of chemically modified silica gels is described. This involves their utilization for the generation of gaseous standard mixtures consisting of methyl chloride as the analyte and nitrogen as a carrier gas to be used for the calibration of the GC-FID system. N-methylmorpholine was chemically bonded to the propylsilylated surface of silica gel forming chloride of an appropriate immobilized compound which, under certain conditions, undergoes thermal decomposition yielding a single, volatile component (methyl chloride). Such a method of generating specific amounts of a standard substance can be used both for a single point calibration and …

Identification Of The Adp-L-Glycero-D-Manno-Heptose-6-Epimerase (Rfad) And Heptosyltransferase Ii (Rfaf) Biosynthesis Genes From Nontypeable Haemophilus Influenzae 2019, W A. Nichols, B W. Gibson, William Melaugh, N G. Lee, M Sunshine, M A. Apicella

Chemistry Faculty Publications

Haemophilus influenzae is an important human pathogen. The lipooligosaccharide (LOS) of H. influenzae has been implicated as a virulence determinant. To better understand the assembly of LOS in nontypeable H. influenzae (NtHi), we have cloned and characterized the rfaD and rfaF genes of NtHi 2019, which encode the ADP-L-glycero-D-manno-heptose-6-epimerase and heptosyltransferase II enzymes, respectively. This cloning was accomplished by the complementation of Salmonella typhimurium lipopolysaccharide (LPS) biosynthesis gene mutants. These deep rough mutants are novobiocin susceptible until complemented with the appropriate gene. In this manner, we are able to use novobiocin resistance to select for specific NtHi LOS inner core …

Characterization Of A Transposon Tn916-Generated Mutant Of Haemophilus Ducreyi 35000 Defective In Lipooligosaccharide Biosynthesis, B W. Gibson, A A. Campagnari, William Melaugh, M A. Apicella, S Grass, Jing Wang, Katherine L. Palmer, R S. Munson

Chemistry Faculty Publications

To define the role of the surface lipooligosaccharide (LOS) of Haemophilus ducreyi in the pathogenesis of chancroid, Tn916 mutants of H. ducreyi 35000 defective in expression of the murine monoclonal antibody (MAb) 3F11 epitope on H. ducreyi LOS were identified by immunologic screening. One mutant, designated 1381, has an LOS which lacks the MAb 3F11 epitope and migrates with an increased mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gene disrupted by the Tn916 element in strain 1381 was identified by cloning the sequences flanking the Tn916 element. The sequences were then used to probe a lambda DASHII genomic library. …

Complexes Between Nascent Polypeptides And Their Molecular Chaperones In The Cytosol Of Mammalian Cells, Daryl K. Eggers, W. J. Welch, W. J. Hansen

Faculty Publications, Chemistry

Folding of newly synthesized proteins in vivo is believed to be facilitated by the cooperative interaction of a defined group of proteins known as molecular chaperones. We investigated the direct interaction of chaperones with nascent polypeptides in the cytosol of mammalian cells by multiple methods. A new approach using a polyclonal antibody to puromycin allowed us to tag and capture a population of truncated nascent polypeptides with no bias as to the identity of the bound chaperones. In addition, antibodies that recognize the cytosolic chaperones hsp70, CCT (TRiC), hsp40, p48 (Hip), and hsp90 were compared on the basis of their …

Sodium Nitrite Alone Protects The Brain O _I_ Microsomal Ca -Atpase Against Potassium Cyanide-Induced Neurotoxicity In Rats, Odutayo O. Odunuge, G.. A. Adenuga

Faculty Publications

The effect of a short-term oral administration of potassium cyanide (KCN) (200 ppm in diet) with or without sodium nitrite (NaNO2) pretreatment on rat brain microsomal Ca2* ATPase was investigated. The specific activity value of the enzyme significantly decreased (p<0.05) by 50% compared with control and by 63% for KCN-treated rats compared with KCN-treated rats pretreated with NaNO;,. There was no significant difference at the h = 0.05 level between the values obtained for the control and KCN-treated rats pretreated with NaNO,. These results show both that feeding lowers brain microsomal Ca2f-ATPase activity and that NaNO, has a protective role (antidote function) in that respect.

Molecular Engineering Of Side-Chain Liquid Crystalline Polymers By Living Polymerizations, Coleen Pugh, Alan L. Kiste

Chemistry and Biochemistry

“Living” anionic, cationic, metalloporphyrin and ring-opening metathesis polymerizations have been used to prepare well-defined side-chain liquid crystalline homopolymers, block and graft copolymers and statistical copolymers. This paper analyzes their successes and failures by reviewing the mechanistic aspects and experimental conditions of each type of polymerization, and identifies other classes of mesogenic monomers that could be polymerized in a controlled manner in the future. The emerging structure/property relationships are then identified using well-defined SCLCPs in which only one structural feature is varied while all others remain constant.

The thermal transitions of liquid crystalline polymethacrylates, polynorbomenes and poly(viny1 ether)s reach their limiting …

Protein-Protein And Protein-Dna Interactions At The Bacteriophage T4 Dna Replication Fork. Characterization Of A Fluorescently Labeled Dna Polymerase Sliding Clamp, Daniel J. Sexton, Theodore E. Carver, Anthony J. Berdis, Stephen J. Benkovic

Chemistry Faculty Publications

The T4 DNA polymerase holoenzyme is composed of the polymerase enzyme complexed to the sliding clamp (the 45 protein), which is loaded onto DNA by an ATP-dependent clamp loader (the 44/62 complex). This paper describes a new method to directly investigate the mechanism of holoenzyme assembly using a fluorescently labeled cysteine mutant of the 45 protein. This protein possessed unaltered function yet produced substantial changes in probe fluorescence intensity upon interacting with other components of the holoenzyme. These fluorescence changes provide insight into the role of ATP hydrolysis in holoenzyme assembly. Using either ATP or the non-hydrolyzable ATP analog, adenosine …

The Carboxyl Terminus Of The Bacteriophage T4 Dna Polymerase Is Required For Holoenzyme Complex Formation, Anthony J. Berdis, Patrice Soumillion, Stephen J. Benkovic

Chemistry Faculty Publications

To further elucidate the mechanism and dynamics of bacteriophage T4 holoenzyme formation, a mutant polymerase in which the last six carboxyl-terminal amino acids are deleted, was constructed, overexpressed, and purified to homogeneity. The mutant polymerase, designated ΔC6 exo−, is identical to wild-type exo− polymerase with respect to kcat, kpol, and dissociation constants for nucleotide and DNA substrate. However, unlike wild-type exo− polymerase, the ΔC6 exo− polymerase is unable to interact with the 45 protein to form the stable holoenzyme. A synthetic polypeptide corresponding to the carboxyl terminus of the wild-type exo− polymerase was tested as an in vitro inhibitor of …