Biochemistry Commons^™

Gene Synthesis, Cloning, Expression, Purification And Biophysical Characterization Of The C2 Domain Of Human Tensin, Kiran Sukumar Gajula

Doctoral Dissertations

Tensin is a large "docking" protein found in the adhesive junctions of animal cells and recruited early in the development of cell-substrate contacts. There it binds to the cytoplasmic domain of integrin β1 and caps the barbed ends of filamentous actin. This forms a rational basis for its implication in a direct role in the mechanics of membrane-cytoskeleton interactions. Tensin provides a physical link between the actin cytoskeleton, integrins, and other proteins at the cell-substrate contacts. Its overall biochemical properties are a function of its domain composition and architecture, i.e., the domains that are present and their relative positions in …

Layer-By-Layer Self -Assembly For Enzyme And Dna Encapsulation And Delivery, Amish Patel

Doctoral Dissertations

Thin wall microcapsules were formed via Layer-by-Layer Self-Assembly of alternate adsorption of oppositely charged polyelectrolyte on microcores. After the core dissolution, empty polymeric shells with 20–25 nm thick walls were obtained. These microcapsules were loaded with Myoglobin, Hemoglobin and Glucose Oxidase by opening capsule pores at low pH and closing them at higher pH. The native structure of the enzyme was not affected due to different treatments. Biocompatible nanoshells were also prepared for encasing DNA. Using the same Layer-by-Layer Self-Assembly approach nanoparticle were constructed containing DNA as one of the layers. The nanoparticles of different architecture were used to deliver …

Stimulation Of Capacitative Calcium Entry In Hl-60 Cells By Nanosecond Pulsed Electric Fields, Jody A. White, Peter F. Blackmore, Karl H. Schoenbach, Stephen J. Beebe

Bioelectrics Publications

Nanosecond pulsed electric fields (nsPEFs) are hypothesized to affect intracellular structures in living cells providing a new means to modulate cell signal transduction mechanisms. The effects of nsPEFs on the release of internal calcium and activation of calcium influx in HL-60 cells were investigated by using real time fluorescent microscopy with Fluo-3 and fluorometry with Fura-2. nsPEFs induced an increase in intracellular calcium levels that was seen in all cells. With pulses of 60 ns duration and electric fields between 4 and 15 kV/cm, intracellular calcium increased 200-700 nM, respectively, above basal levels (similar to100 nM), while the uptake of …

Protocols For Disease Classification From Mass Spectrometry Data, Michael Wagner, Dayanand Naik, Alex Pothen

Mathematics & Statistics Faculty Publications

We report our results in classifying protein matrix-assisted laser desorption/ionizationtime of flight mass spectra obtained from serum samples into diseased and healthy groups. We discuss in detail five of the steps in preprocessing the mass spectral data for biomarker discovery, as well as our criterion for choosing a small set of peaks for classifying the samples. Cross-validation studies with four selected proteins yielded misclassification rates in the 10-15% range for all the classification methods. Three of these proteins or protein fragments are down-regulated and one up-regulated in lung cancer, the disease under consideration in this data set. When cross-validation studies …

Urea Hydrolysis Using Immobilized Urease, Anita Arullan

Student Works (2000-2009)

An inexpensive carrier matrix. vermiculite was used for immobilizing urease. A cost effective and simple method of coupling was developed using vermiculite. Immobilization of urease on Amberlite MB-land Nylon-6 tube were also studied. Activities of free and immobilized urease were studied and compared: Among all methods used for immobilization of urease. the method of coupling the enzyme to vermiculite gave the highest retained activity of 82% (Method I) and 89% (Method 2) at pH 6.0 and 5.5 respectively. Urease immobilized by Method I retained 69% and 30% of its original activity and Method 2 (using glutaraldehyde as coupling agent) retained …

Evaluation Of Toxicity, Bioavailability And Speciation Of Lead, Zinc And Cadmium In Mine/Mill Wastewaters, Mujde Erten-Unal, Bobby G. Wixson, Nord Gale, Jerry L. Pitt

Civil & Environmental Engineering Faculty Publications

The toxicity of common compounds of lead, cadmium and zinc was evaluated in waters similar to that found in the world's largest lead producing area in Missouri. Static, acute toxicity tests were performed using fathead minnows (Pimephales promelas) and water fleas (Daphnia magna) respectively. Test organisms were subjected to varying amounts of sulfide, carbonate, chloride and sulfate salts of lead, zinc and cadmium mixed in hard, alkaline waters typical to this region. Median lethal concentrations were calculated using nominal versus measured metal concentrations. Measured metal concentrations included four different metal fractionation (extraction/filtration) techniques at different pH …

Longitudinal Study Of Plasma And Erythrocyte Selenium Levels In Type I Diabetic Children During Development And Treatment Of Ketoacidosis, Pi-Hsia Fan

Chemistry & Biochemistry Theses & Dissertations

Most studies on the biological activity of selenium have focused on its role at the active site in the ubiquitous enzyme, glutathione peroxidase (GSH-Px) which catalyzes the removal of hydrogen peroxide and organic hydroperoxides (1). A connection between selenium deficiency and a wide range of diseases in animals and humans has been reviewed (2). Development of various forms of human cancer (3), failure of the immune system (4), as well as some cardiovascular diseases (5) have been reported or suspected to correlate with selenium deficiency. However, the physiological function of selenium in these different diseases is not fully understood, and …

Structural Characterization Of Fucoidan And Its Role In The Hemizona Assay, Manish S. Patankar

Chemistry & Biochemistry Theses & Dissertations

Fucoidan is a polysaccharide commonly found in brown algae. It is a sulfated polymer of the monosaccharide L-fucose. Fucoidan is thought to function in water retention by algae. Commercially available fucoidan is extracted from a brown algae called Eurus vesiculosus.

This polysaccharide has immense potential as a biologically active agent: (i) it exhibits anticoagulant action, (ii) it has also been reported to block retroviral replication and (iii) it is capable of blocking sperm-egg binding in sea urchin, mouse and rabbit. Recently, fucoidan has also been shown to block the interaction of human sperm and egg in the hemizona assay …

Purification And Characterization Of The Prostate Cancer-Associated Antigen: 7e11-C5, Qi Feng

Theses and Dissertations in Biomedical Sciences

A murine monoclonal antibody (MAb), 7E11-C5, recognizes a prostate organ-specific antigen. Under reducing conditions, the 7E11-C5 antigen exhibited major 100-Kd and minor 70-Kd bands, suggesting that the antigen comprises two peptide chains. The antigen was detected in pooled normal human seminal plasma, xenograft tissue extract of a prostate adenocarcinoma cell line, LNCaP, and tissue extracts of benign prostatic hyperplasia (BPH) and prostate carcinomas, but was not identified in any of the non-prostate tissue extracts tested. Antibodies of the three major prostate-associated antigens (prostatic acid phosphatase (PAP), prostate specific antigen (PSA) and prostate secretory protein (PSP)) were unable to block the …

Metabolism Of Arachidonate-Containing Phospholipid Molecular Species In The Murine Macrophage-Like Cell Line, P388d1, Crystal R. Waites

Electronic Theses and Dissertations

Glycerophospholipids of mammalian cells exist as chemically diverse structures with various fatty acids at the sn-1 and sn-2 positions. Arachidonic acid, a polyunsaturated fatty acid, which may be converted to biologically active eicosanoids such as prostaglandins, thromboxanes, and leukotrienes, is found predominantly in the sn-2 position of glycerophospholipids. The purpose of this study was to examine, at the level of the individual molecular species, the incorporation of arachidonate into phospholipids and its release from phospholipids during stimulation. In this way, the specificity of the enzymes controlling arachidonate metabolism could be examined in order to clarify the processes that control the …

Development Of An On-Line Computer System For Enzyme Kinetics Studies, Jacqueline R. Thweatt

Chemistry & Biochemistry Theses & Dissertations

The PDP-11/10 minicomputer was interfaced to the Beckman 25 ultraviolet/visible spectrophotometer and this on-line computer system was applied to enzyme kinetics studies. This interface involved hardware equipment to locate, amplify, and transmit the analog signal to an analog-to-digital converter, and software to acquire and store the digital data. Data processing programs were used to calculate kinetic parameters and statistical information from the acquired data.

The Varian Cary 219 ultraviolet/visible spectrophotometer can be readily interfaced to the Apple II Plus microcomputer by hardware equipment which can be purchased from the manufacturer. Various software may also be purchased for use in enzyme …

Report On Wastewater Disposal To Board Of County Commissioners, Clark County Nevada, Clair N. Sawyer

Publications (WR)

This report is concerned with the management of waste-waters and subsurface drainage from the City of Las Vegas and its environs and the BMI complex, all in Clark County, Nevada. The prime objective is to recommend a plan of action which will utilize the natural resources of the area in the least costly manner and still protect Lake Mead for recreational purposes and use as a public water supply, At this writing, conditions in the upper Las Vegas arm of Boulder Basin are quite unsatisfactory for some recreational purposes due to the extensive blooms of algae which develop. Experience at …

Comprehensive Survey Of Sedimentation In Lake Mead, 1948-49, W. O. Smith, C. P. Vetter, G. B. Cummings, U.S. Bureau Of Reclamation

Publications (WR)

Reservoirs are becoming an increasingly prominent feature of the American landscape. Built for flood mitigation and to change a fluctuating river into a dependable source of water for irrigation, power, and other purposes, they are predestined, like natural lakes, to be destroyed sometime following their creation. Sedimentation sooner or later robs most lakes and reservoirs of their capacity to store water. The significance of sedimentation in the life of Lake Mead, the largest artificial reservoir in the world, was realized when the plan for the reservoir was conceived, and an aerial survey of the floor was made in 1935 before …