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Biomedical subjects

M D Morris

Publications and source records attributed to M D Morris.

At least 19 recordsLinked to original sources

Acrylamide polymerization kinetics in gel electrophoresis capillaries. A Raman microprobe study.

The formation of 3.5% T, 3.3% C cross-linked polyacrylamide is monitored in 75-microns-i.d. electrophoresis capillaries by Raman microprobe spectroscopy. The disappearance of the acrylamide 1292-cm-1 band is followed with 60-s time resolution for 30 min, and 2-4 min resolution for up to 10 h. Polymerization is 98% complete in 1.5 h and greater than 99% complete after 2 h. In the 900-1700-cm-1 region no bands attributable to cross-linking are observable. Reaction in the capillary follows second-order kinetics. The reaction is faster in the bulk system because heat dissipation is not sufficient to maintain a constant temperature.

Acrylamide

Noninvasive imaging of nanogram quantities of DNA in agarose electrophoresis gels.

Image processing techniques which separate the DNA Kerr effect and induced electrokinetic distortion contributions to electric birefringence images of agarose nucleic acid electrophoresis gels are described. Under standard electrophoresis conditions, detection limits of 10 ng of DNA per well are obtained in hydroxyethylated agarose without signal averaging or 7.5 ng with averaging of four measurements. Maintaining constant gel temperature is shown to improve the quality of the images. Monochromatic light (589 nm) is shown to give a small increase in sensitivity compared to polychromatic (650 +/- 20 nm) light.

DNA

Effects of analyte velocity modulation on the electroosmotic flow in capillary electrophoresis.

Modulation of the electroosmotic flow in capillary zone electrophoresis by modulation of the driving voltage gives rise to a flow profile that changes between laminar and flat profiles. The changing flow profile induces a radial movement of sample species to and from the capillary surface. The induced sample concentration gradient can be monitored by carefully probing the capillary surface. The resulting signal is a derivative of the normal-shaped peak. Derivative-shaped peaks can be observed with cations, but not with anions. Anions are unable to approach the double-layer region and therefore are unaffected by the modulation process.

Acetonitriles

Electrokinetic and Kerr effect contributions to electric birefringence images of agarose electrophoresis gels.

Multilinear regression analysis is used to test a model (Lanan et al., 1991) for electric birefringence imaging (EBI) of DNA fragment bands in agarose electrophoresis gels. EBI signals attributed to localized electro-kinetic gel distortion and to intrinsic DNA birefringence are studied by fitting ethidium bromide fluorescence profiles to EBI results. Fluorescence polarization imaging (FPI) is used to assess the influence of localized gel distortion on nucleic acid orientation across a fragment band. It is shown that DNA aligns parallel, on average, with an applied electric field independent of its location within a band. DNA orientation varies as the 1.2 power in electric field strength under the conditions tested.

Birefringence

On-line multichannel Raman spectroscopic detection system for capillary zone electrophoresis.

An on-line multichannel Raman spectroscopic detection system for capillary zone electrophoresis using a charge-coupled device as the detector is described. Resonant, near-resonant and non-resonant excitation Raman spectroscopies are employed. The 400 cm-1 spectral window provides adequate information to identify resolved and unresolved compounds. The use of analyte velocity reduction to allow increased data acquisition times is described. With near-resonant enhancement, the technique is shown to allow detection of 500 attomoles of methyl orange from a 5-nl injection volume, corresponding to 1 x 10(-7) M.

Electrophoresis

Resonance Raman spectroscopy of bilirubins: band assignments and application to bilirubin/lipid complexation.

Resonance Raman spectra of bilirubins IX alpha, III alpha, and XIII alpha and mesobilirubin XIII alpha in alkaline aqueous and chloroform solutions are reported. Partial band assignments of bilirubin IX alpha are proposed. The model compounds confirm assignments of bands of the Raman spectrum of bilirubin IX alpha to each of the two different pyrromethenones. Resonance Raman spectra of mesobilirubin IV alpha, vinylneoxanthobilirubinic acid, and vinylisoneoxanthobilirubinic acid in alkaline aqueous solution and of the tetra-n-butylammonium salt of bilirubin IX alpha are used to define markers for the presence or absence of internal hydrogen bonds. Interaction of bilirubin dianion and sphingomyelin liposomes is studied. The Raman evidence suggests that in the bilirubin dianion/liposome complex the intramolecular hydrogen bonds between the propionate groups and the lactam NH/CO are ruptured. It is proposed that in the complex the bilirubin propionates form ion pairs with the quaternary ammonium ion of the choline moiety of sphingomyelin.

Bilirubin

Electric birefringence imaging of electrokinetic agarose orientation.

Time-resolved and steady-state electric birefringence imaging with a slow-scan video camera is used to study orientation during DNA agarose gel electrophoresis. The hydrodynamically induced gel distortion is shown to be the major source of birefringence under electrophoresis running conditions and to generate a birefringence image that approximates the image of the DNA concentration gradient in the electric field direction. A fluid kinematic model is presented to describe the spatial distribution of steady-state birefringence and is verified with fluorescence measurements of DNA distribution. The stress-optic coefficient of 1% agarose gel is measured by mechanical compression and used to evaluate the magnitude of the induced strain on the gel during electrophoresis.

Birefringence

A mathematical model for radiation-induced myelopoiesis.

A model for damage, repair, killing, and repopulation of myelopoietic marrow is presented. Evaluation produces time and dose-rate profiles during and following any complex irradiation. Equations model variable dose rates, multiple exposures, different sources, and arbitrary intervals between treatments. If factors which dominate the control of biological processes can be demonstrated, an option is to set biological rate constants to experimentally determined values. Previously, knowledge did not permit identification of dominating biological processes and their temporal rates. But a unique feature of this study is that unspecified lesions for killing and injury of cells are evaluated from mortality data on the animal species of choice. "Unspecified" is used to indicate a condition of assumption-free modeling of molecular processes, whereby rate constants for cellular effects are simply computed directly from animal mortality data. Coefficients (estimated by maximum-likelihood methods for nonspecific processes) are compared with experimental values for specific processes. The model has many uses, including modeling of the myelopoietic potential as a function of time. Another option is to calculate the whole-body survival curve for cells that control myelopoiesis as a result of the treatment schedule. Also through simple extensions of the model, an extremely complex protocol can be identified with an equivalent prompt dose value--even for partial-body, fractionated exposures.

Animals

Estimation of coefficients in a model of radiation-induced myelopoiesis from mortality data for mice following X-ray exposure.

The rate coefficients in the model of cell kinetics and mortality introduced by Jones et al. (Radiat. Res. 128, 258-266 (1991)) are estimated using mortality data from several mouse experiments. The evaluated model fits data from a large variety of prompt, protracted, and fractionated irradiations with 250-k Vp X rays with good fidelity. Although the maximum-likelihood estimates are not unique, all estimates lead to greater cell survival than that observed in in vitro experiments on nonterminally differentiated reproducing cells from the marrow.

Animals

Hadamard transform photothermal deflection densitometry of electrophoretically blotted proteins.

Hadamard transform spatial multiplexing techniques are applied to laser densitometry in order to prevent the photo-induced degradation of sensitive materials. Photochemical and thermal degradations can often occur in point focused scanning laser densitometry. In spatial multiplexing, the excitation source is defocused and efficiently distributed throughout the sample, reducing local power density. In this paper, we describe the application of Hadamard transform spatial multiplexing to transverse photothermal deflection spectroscopy (PDS). Proteins western blotted on nitrocellulose membrane are line imaged using the Hadamard transform PDS densitometer. For comparison, the blots are imaged with a high-dynamic-range video densitometer.

Alkaline Phosphatase

Surface-enhanced Raman scattering of dopamine at polymer-coated silver electrodes.

Attenuation of dopamine surface-enhanced Raman scattering (SERS) spectra at silver electrodes by protein adsorption is described. Polymer-modified electrodes eliminate protein adsorption effects. Partially hydrolyzed cellulose acetate coatings prevent protein adsorption and allow transport of dopamine to and from the electrode surface. Dopamine SERS spectra from these electrodes are similar to those obtained at uncoated electrodes. Perfluorosulfonate (Nafion) coatings also prevent protein adsorption. However, dopamine transported to the electrode remains trapped inside the coating and cannot be removed.

Animals

Statistical approach to combining the results of similar experiments, with application to the hematologic effects of extremely-low-frequency electric field exposures.

A large proportion of scientific effort in investigating the possible biological effects of exposure to extremely-low-frequency (ELF) fields consists of laboratory studies on experimental animals. Most experiments in which hematologic properties are measured show no statistically significant effect due to exposure. However, some studies show significant effects which, in general, are not clearly reproducible. A difficult question must then be addressed: Are these relatively few indications of ELF effects statistical artifacts due to the increased risk of a type I error in multiple studies, or is there a real biological effect that is undetected in most studies due to the relatively small sample sizes commonly used? A statistical approach for examining the accumulated results of multiple experiments which results in a single test for treatment effect is presented. The technique requires very mild assumptions, and is valid for experiments that vary widely in specific characteristics such as exposure level, duration, and laboratory. The method is applied to the results of a collection of hematologic and serum chemistry experiments, and the combined results indicate the existence of experimental effects on some end points.

Animals

Hematopoietic death of unprotected man from photon irradiations: statistical modeling from animal experiments.

Most estimates of the radiation dose lethal to 50 per cent of a human population are based on historical data taken from well-known experiences reflecting inadequately known physical and biological conditions, or from medical procedures where individual patients received advantages of modern clinical care. It has been debated as to whether the experience of unprotected man would more closely reflect that of hospital patients or of the radiobiological studies with large animals. The issue at question is whether the apparent two-fold or more increased susceptibility of large animals to death from bone marrow damage (compared with the majority of estimates for man) is due more to true interspecies differences or the lack of medical support. This study is an attempt to assess the radiosensitivity of unprotected man in terms of the composite animal experiments. Based upon an extensive data base containing 121 separate animals studies using 13 different species, an estimate of the mortality dose-response relationship due to a uniform, continuous field of photon radiation is predicted for 70 kg unprotected man. Man is assumed to have a level of radiation sensitivity similar to that of the species represented in the data base, after adjustment for body weight. The mathematical model used includes fixed terms to account for effects of body weight and dose rate, and random terms reflecting inter- and intra-species variation and experimental error. Point predictions and 95 per cent prediction intervals are given for the LD05, LD10, LD25, LD50, LD75, LD90, and LD95, for dose rates ranging from 0.01 to 0.5 Gy/min, and treatment times ranging from about 2 min to about 24 h. At 1 cGy/min our point prediction of the LD50 is 299 cGy with an associated 95 per cent prediction interval of (168 cGy, 535 cGy). The analogous values at 50 cGy/min are 183 cGy and (103 cGy, 326 cGy).

Animals