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Biomedical subjects

B R Locke

Publications and source records attributed to B R Locke.

14 recordsLinked to original sources
PubMed2006

The roles of ozone and zeolite on reactive dye degradation in electrical discharge reactors.

In this study high voltage pulsed corona electrical discharge advanced oxidation processes (AOPs) were applied to bleach and degrade C.I. Reactive Green 8 and C.I. Reactive Red 45 organic dyes in water solutions. Two types of hybrid gas/liquid high voltage electrical discharge (corona) reactors, known as hybrid series and hybrid parallel were studied. The difference between these reactors relates to electrode configuration, which affects the amounts of ozone, hydrogen peroxide and hydroxyl radicals produced. Experiments were conducted using dye concentrations of 20 mgl(-1) and 75 mgl(-1), with and without NH4ZSM5 zeolite addition in order to determine possible effects of added solid particles to total process efficiency. The role of ozone in combination with zeolites was assessed through comparative direct ozonation experiments with ozone supplied by an ozone generator. UV/VIS spectrophotometric measurements and measurements of total organic carbon (TOC) were used for the determination of decolorization and mineralization rates.

Coloring Agents↗
PubMed2004

The removal of Direct Orange 39 by pulsed corona discharge from model wastewater.

Untreated wastewater from the dye industry and dyehouses cannot be directly discharged into the environment due to the high content of organic matter and intensive colouration, even with low concentrations of dye. In this paper, the application of a high voltage pulsed electrical discharge in the aqueous phase has been assessed for the dye degradation. Experiments were conducted in a batch reactor using model wastewater of the commercial water-soluble monoazo dye C.I. Direct Orange 39 (DO39). The effects of zeolite and ferrous sulphate in combination with the corona discharge were examined. Experiments were conducted for a range of process parameters including pH, conductivity, type and amount of zeolite, and ferrous sulphate concentration. A mathematical model to describe the kinetics of DO39 degradation in the corona reactor was developed. Aqueous phase pulsed streamer corona discharge as a method for coloured wastewater treatment showed very high effectiveness in the case of iron salt addition (Fenton's reaction). Low pH enhanced dye removal by corona in the absence of zeolite, thus implying that the acid properties of zeolites are important in dye degradation. Ecological parameters such as COD, TC, IC, TOC and IC50 measured before and after corona treatment showed that the treated wastewater can be discharged into the environment or reused as process water.

Azo Compounds↗
PubMed1999

Analysis of cell growth kinetics and substrate diffusion in a polymer scaffold.

The cultivation of cartilage cells (chondrocytes) in polymer scaffolds leads to implants that may potentially be used to repair damaged joint cartilage or for reconstructive surgery. For this technique to be medically applicable, the physical parameters that govern cell growth in a polymer scaffold must be understood. This understanding of cell behavior under in vitro conditions, where diffusion is the primary mode of transport of nutrients, may aid in the scale-up of the cartilage generation process. A mathematical model of chondrocyte generation and nutrient consumption is developed here to analyze the behavior of cell growth in a biodegradable polymer matrix for a series of different thickness polymers. Recent literature has implied that the diffusion of nutrients is a major factor that limits cell growth (Freed et al., 1994). In the present paper, a mathematical model is developed to directly relate the effects of increasing cell mass in the polymer matrix on the transport of nutrients. Reaction and diffusion of nutrients in the cell-polymer system are described using the fundamental species continuity equations and the volume averaging method. The volume averaging method is utilized to derive a single averaged nutrient continuity equation that includes the effective transport properties. This approach allows for the derivation of effective diffusion and rate coefficients as functions of the cell volume fraction. The cell volume fraction as a function of time is determined by solution of a material balance on cell mass. Growth functions including the Moser, a modified Contois, and an nth-order heterogeneous growth kinetic model are evaluated through a parameter analysis, and the results are compared to experimental data found in the literature. The results indicate that cellular functions in conjunction with mass transfer processes can account partially for the general trends in the cell growth behavior for various thickness polymers. The Contois growth function appeared to describe the data more accurately in terms of the lag period at early times and the long time limits. However, all kinetic growth functions required variations in the kinetic parameters to fully describe the effects of polymer thickness. This result implies that restricted diffusion of nutrients is not the sole factor limiting cell growth when the thickness of the polymer is changed. Therefore, further experimental data and model improvements are needed to accurately describe the cell growth process.

Animals↗
PubMed1999

Effects of spatial variation of cells and nutrient and product concentrations coupled with product inhibition on cell growth in a polymer scaffold.

The effects of spatial variation of cells and nutrient and product concentration, in combination with product inhibition in cell growth kinetics on chondrocyte generation in a polymer scaffold, are analyzed. Experimental studies reported previously have demonstrated spatial dependence in the cultivation of chondrocytes. In the present study, the cell-polymer system is assumed to consist of two distinct phases. The cells, fluid, polymer matrix, and extracellular matrix comprise one phase, and the other phase consists of a fluid and polymer matrix. The only two species in the fluid considered to affect cell growth are the nutrient and product. The multiphase transport process of these two species in the cell-polymer system is described by the species continuity equations and corresponding boundary conditions for each individual phase. A volume-averaging approach is utilized for this system to derive averaged species continuity equations for the nutrient and product concentrations. The volume-averaging approach allows for a single species in a two-phase system to be represented by a single averaged continuity equation. Competitive product inhibition, saturation kinetics of substrate, and cell population control are assumed to affect the cell growth kinetics. A modified Contois growth kinetic model is used to represent the three factors that affect cell growth. A parameter analysis is performed and the results are compared qualitatively with experimental data found in the literature.

Animals↗
PubMed1999

Diffusional anisotropy is induced by subcellular barriers in skeletal muscle.

The time- and orientational-dependence of phosphocreatine (PCr) diffusion was measured using pulsed-field gradient nuclear magnetic resonance (PFG-NMR) as a means of non-invasively probing the intracellular diffusive barriers of skeletal muscle. Red and white skeletal muscle from fish was used because fish muscle cells are very large, which facilitates the examination of diffusional barriers in the intracellular environment, and because they have regions of very homogeneous fiber type. Fish were cold-acclimated (5 degrees C) to amplify the contrast between red and white fibers. Apparent diffusion coefficients, D, were measured axially, D(axially) and radially, D(radially), in small muscle strips over a time course ranging from 12 to 700 ms. Radial diffusion was strongly time dependent in both fiber types, and D decreased with time until a steady-state value was reached at a diffusion time approximately 100 ms. Diffusion was also highly anisotropic, with D(axially) being higher than D(radially) for all time points. The time scale over which changes in D(radially) occurred indicated that the observed anisotropy was not a result of interactions with the thick and thin filament lattice of actin and myosin or restriction within the cylindrical sarcolemma, as has been previously suggested. Rather, the sarcoplasmic reticulum (SR) and mitochondria appear to be the principal intracellular structures that inhibit mobility in an orientation-dependent manner. This work is the first example of diffusional anisotropy induced by readily identifiable intracellular structures.

Animals↗
PubMed1999

When can the Ogston-Morris-Rodbard-Chrambach model be applied to gel electrophoresis?

The Ogston-Morris-Rodbard-Chrambach theory of gel electrophoresis is consistent with predictions from the volume averaging method with respect to the equivalence of the accessible volume fraction to the ratios of gel mobility to free solution mobility and the gel diffusion coefficient to free solution diffusion coefficient for the limiting case of small molecule electrophoresis with low electrical fields, low gel concentrations, and nonconductive gel fibers. When these conditions are not valid, more extensive calculations are required to determine the mobility and diffusion coefficient ratios as functions of the geometry and electrical field within the gel. The volume averaging theory shows that it is important to account for the electrical conductivity properties of the fibers that make up a gel electrophoresis medium, and this aspect is consistent with early theories of transport phenomena in gel electrophoresis.

Electrophoresis↗
PubMed1999

Transdermal water mobility in the presence of electrical fields using MR microscopy.

Magnetic resonance microscopy of skin from hairless rats under the influence of electrical fields was conducted for two cases: 1) low voltage constant electrical fields and 2) high-voltage short pulse, electrical fields. Under conditions of the low voltage and low current iontophoresis, i.e., 0 to 20 V, and 0 to 0.5 mA/cm2, it was found that the skin structure, as observed by magnetic resonance microscopy, did not significantly change until 20 Volts were applied across the 0.1 cm thick skin. Under these conditions, the viable epidermis appeared to swell, and this result corresponded to observations from scanning electron microscopy and other research from the literature. High voltage electrical fields, i.e., 220 V 1 ms pulses repeated once per second, appeared to hydrate the stratum corneum as is consistent with published literature on electroporation. In the case of iontophoresis, water self-diffusion coefficients in the epidermis and hair follicle regions at all voltages were affected by the electrical field. Statistical analysis at the 95% confidence level for the comparison of the average differences between diffusion coefficients with the electrical field on and with the electrical field off for pair matched pixels for the viable epidermis show that for 5 V (p = 0.00377), 10 V (p = 0.0108), 20 V (p = 0.0219) regimes there are statistically significant (p < or = 0.05) changes due to the applied electric field. The same analysis for the hair follicle region at 5 V (p = 6.89 x 10(-7)), 10 V (p = 1.42 x 10(-5)), 20 V (p = 3.23 x 10(-3)) also show statistically significant changes (p < or = 0.05). When the electroporation pulse was applied, the water diffusion coefficients increased by about 30% to 6.6 x 10(-6) cm2/s +/- 2.4 x 10(-7) cm2/s and 8.3 x 10(-6) cm2/s +/- 3.7 x 10(-7) cm2/s, for the epidermis and hair follicle regions, respectively. Significant differences were noted between diffusion coefficients in the viable epidermis and the hair follicles for all cases.

Animals↗
PubMed1998

Pluronic copolymer liquid crystals: unique, replaceable media for capillary gel electrophoresis.

Liquid crystalline solutions of Pluronic copolymers are versatile alternatives to solutions of entangled, random coil polymers as replaceable media for capillary gel electrophoresis (CGE). Pluronic copolymers are tri-block polymers of poly(ethylene oxide) [(EO)x] and poly(propylene oxide) [(PO)y] with the general formula (EO)x(PO)y(EO)x. Large micelles form in aqueous solutions in which central, hydrophobic cores of (PO)y segments are surrounded by "brushes" of hydrated (EO)x tails. Solutions of Pluronic F127 (BASF Performance Chemicals) in a concentration range of about 18-30% are liquids at refrigerator temperatures (< or = 5 degrees C) and are easily introduced into capillaries. A self-supporting, gel-like liquid crystalline phase is formed as the temperature is raised to > or = 20 degrees C. This liquid crystalline phase consists of spherical micelles with diameters of 17-18 nm which pack with local cubic symmetry. CGE in Pluronic F127 liquid crystals separates species within several chemical classes as varied as nucleoside monophosphates and organic dyes, oligonucleotides of 4-60 nucleotides, DNA fragments of 50-3000 base pairs (bp), and supercoiled plasmid DNAs of 2000-10,000 bp. Mechanisms of molecular sieving in polymer liquid crystals must differ in fundamental ways from separations in random polymer gels because molecules move around uncrosslinked obstacles that are larger than the smallest dimensions of typical analytes. Molecular sieving in Pluronic liquid crystals is envisioned to occur as molecules squeeze between hydrated (EO)x strands of micelle brushes, or through brushtips and interstitial spaces between micelles. Small molecules such as nucleotides appear to separate by a different mechanism involving partitioning between hydrophilic and hydrophobic environments. This process is termed "hydrophobic interaction electrophoresis". The unique structures of Pluronic copolymers and their liquid crystalline phases provide new challenges and opportunities in separations science.

Crystallography↗
PubMed1998

Electrophoresis in lyotropic polymer liquid crystals.

Excellent electrophoretic separations of a variety of biological molecules can be accomplished by using uncharged, triblock copolymers as the "gel" media. These copolymers form uncrosslinked, lyotropic liquid crystalline phases of large micelles between which molecules must travel. Unlike crosslinked hydrogels in common use, these alternative media have highly ordered internal structures. Pluronic F127, representative of the copolymer class, contains poly(ethylene oxide) (EO) and poly(propylene oxide) (PO) units with an approximate molecular formula (EO)106(PO)70(EO)106. Concentrated (18-30%) solutions of Pluronic F127 are freely flowing liquids at low temperature (0-5 degrees C) but form gel-like, cubic liquid crystals of large, spherical micelles when warmed. The utility of these media is illustrated by separations of linear, double-stranded DNA up to 3,000 bp long by conventional electrophoresis, and of single-stranded DNAs from 4 to 60 nt long by capillary electrophoresis. Extraordinary separations of supercoiled DNAs were also obtained by capillary electrophoresis. The versatility, availability, and ease of use of Pluronic polymers offer major advantages over conventional media for preparative and high performance analytical separations of nucleic acids and other biomolecules. Mechanisms of molecular transport and separation operating in polymer liquid crystals must differ in fundamental ways from those in crosslinked gels. Lyotropic polymer liquid crystals are unique systems for elucidating mechanisms of macromolecule migration in ordered, dense media, and provide opportunities in separations science.

Chemical Phenomena↗
PubMed1997

Spatial resolution of transdermal water mobility using NMR microscopy.

High resolution images were obtained using high-field nuclear magnetic resonance microscopy of in vitro preparations of hydrated hairless rat skin. The major anatomical features observed were comparable to those seen by electron microscopy and include the stratum corneum, the viable epidermis, sebaceous glands, the cell layers surrounding hair follicles (the outer and inner root sheaths), and regions of subcutaneous fatty deposits. Calculated diffusion maps demonstrated that signal intensity is sufficient to obtain quantitative water mobility data from the viable epidermis and the hair follicle/sebaceous gland regions. Images from skin immersed in D2O clearly distinguish signal contributions that arise from fat from those which arise from water, and indicate that the calculated diffusion maps include only proton mobility from water in skin. These results may lead to further applications for using quantitative nuclear magnetic resonance microscopy for examining transdermal transport processes of in vitro skin preparations.

Animals↗
PubMed1997

Reaction-diffusion analysis of the effects of temperature on high-energy phosphate dynamics in goldfish skeletal muscle.

Thermal acclimation results in dramatic changes in the fractional volume of mitochondria within skeletal muscle of teleost fish. We investigated the hypothesis that changes in mitochondrial volume represent a compensatory response to temperature-induced changes in intracellular diffusion coefficients (D) of the high-energy phosphate compounds ATP and creatine phosphate (PCr). Using 31P nuclear magnetic resonance spectroscopy, we determined DPCr and DATP in goldfish (Carassius auratus) skeletal muscle at 25 degrees C and 5 degrees C: DPCr was 3.28 +/- 0.18 x 10(-6) cm2s-1 at 25 degrees C and 2.00 +/- 0.90 x 10(-6) cm2s-1 at 5 degrees C: DATP was 2.13 +/- 0.16 x 10(-6) cm2s-1 at 25 degrees C and was estimated to be 1.30 x 10(-6) cm2s-1 at 5 degrees C. There was no evidence for an effect of acclimation temperature or fiber type on DATP or DPCr. A mathematical reaction-diffusion model was used to calculate profiles of [ATP], [PCr] and the free energy of ATP hydrolysis (delta GATP) in activated goldfish muscle fibers at 5 degrees C and 25 degrees C. The results showed spatial and temporal constancy of [ATP], [PCr] and delta GATP in red fibers at both temperatures, regardless of changes in acclimation temperature or mitochondrial density. The model also showed spatial and temporal constancy of [ATP] in white fibers at 5 degrees C and 25 degrees C, but gradients in [PCr] and delta GATP developed in white fibers under all conditions of temperature and acclimation temperature. These gradients were attenuated in cold-acclimated animals by cold-induced increases in mitochondrial density. However, the model shows that the proximal stimulus for temperature-induced changes in mitochondrial volume density in muscle is not a disruption in intracellular diffusion of high-energy phosphates.

Acclimatization↗
PubMed1996

The effects of temperature, pH, and magnesium on the diffusion coefficient of ATP in solutions of physiological ionic strength.

Intracellular diffusive transport of adenosine triphosphate (ATP) is critical to cellular metabolism. Physical models predict that diffusion coefficients (D) of small molecules are functions of temperature and viscosity of the diffusive environment. Therefore, changes in body temperature, commonly experienced by poikilotherms, are expected to result in changes in the rate of intracellular ATP transport. However, it has been postulated that changes in the electrical charge of ATP may influence the interaction between ATP and the cytosol and that the temperature sensitivity of DATP may deviate from the predicted relationship. To investigate the effects of changes in electrical charge on the temperature sensitivity of DATP, we measured DATP under various conditions of temperature, pH, and pMg2+. Changes in pH and pMg2+ were used to alter the net charge of ATP, and DATP was measured in solutions of physiological ionic strength. Results showed a positive correlation between DATP and temperature; DATP = 1.75 +/- 0.09, 3.68 +/- 0.14, and 4.64 +/- 0.13 (mean +/- S.E.M.) x 10(-6) cm2/s at 5 degrees C, 25 degrees C, and 40 degrees C, respectively. Changes in pH and pMg2+ did not significantly influence DATP, and the change in DATP with respect to temperature was similar to that predicted on the basis of changes in temperature and viscosity of the aqueous medium.

Adenosine Triphosphate↗
PubMed1996

Protein electrophoresis in polyacrylamide gels with templated pores.

An approach is described for the synthesis of nanostructured hydrogels with defined size channels or pores for separations of biological macromolecules. Polyacrylamide gels (15-20% acrylamide) were cast in the presence of high concentrations (5%-28%) of hydrophilic, macromolecular cosolutes either semirigid, rod-like polyelectrolytes (short fragments of DNA or xanthan) or spherical micelles of sodium dodecyl sulfate (SDS). Polyanionic cosolutes were then removed by a combination of diffusion and electrophoresis. These processes are expected to yield gels with 'templated' channels having dimensions near those of the double helical polymers (diameters approximately 2-3 nm, lengths of 50-200 nm) or pores near the size of the spherical SDS micelles (approximately 4-5 nm). This hypothesis was tested by comparing the relative electrophoretic mobilities of proteins (3400 to 43,000 Da) complexed with SDS on templated and conventional gels. Differences in electrophoretic mobilities were observed between all templated gels and control normal gels, demonstrating that the templating process altered the polyacrylamide network. No evidence was found for phase separation of cosolutes from the polyacrylamide network during polymerization. Templated pores are expected to enhance the mobilities of molecules in a particular size range relative to smaller and larger molecules. Gels templated with DNA or xanthan (8-10% final concentration) exhibited little size selectivity, but selectivity was observed for gels templated with SDS varied with polyacrylamide concentration in a manner consistent with the creation of templated pores approximately the size of SDS micelles and larger than the average pore size in a surrounding polyacrylamide network.

Acrylamide↗