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R F DeGrella

Publications and source records attributed to R F DeGrella.

8 recordsLinked to original sources
PubMed1988

Critical evaluation of the TDx Turbo Nephelometry System.

We critically evaluated the Nephelometry System (TDx Turbo) for the TDx Analyzer (Turbo), as currently used for immunoglobulins (IgA, IgG, and IgM) and transferrin assays. We describe hardware, software, reagent, and procedural changes made since the original publication (Clin Chem 1985;31:1474) and the mathematical treatment of data. The unit-dose format of the reagents allows for batch or panel testing of the analytes, with a throughput of 40 tests per hour. Measurements are made at pseudo-equilibrium, with sample-blank correction. The formulation of reagents and the assay scheme are such as to eliminate the need to check these assays for antigen excess. Assay sensitivities are 3% of the standard curve ranges. CVs are 8% for all assays. The system performance, when compared with rate nephelometry and radial immunodiffusion, yielded correlation coefficients of 0.97 or better for the analytes.

Humans↗
PubMed1985

Uptake of fatty acids by isolated heart myocytes.

An adult rat heart myocyte preparation was used to study the uptake and metabolism of the 1-14C-labelled fatty acids decanoate, laurate, myristate, and palmitate. Metabolism was saturable, but free fatty acid accumulation was not. Use of metabolic inhibitors indicated that metabolism and not uptake was rate limiting at 37 degrees C and that uptake was energy dependent. Competition was demonstrated for the metabolic pathways but not for free fatty acid accumulation. The activation energy for uptake decreased with increasing fatty acid chain length. A diffusion mechanism for long chain fatty acid uptake is proposed.

Animals↗
PubMed1985

A nephelometry system for the Abbott TDx analyzer.

A self-contained light-scattering accessory carousel has been developed for the Abbott TDx fluorescence polarization analyzer, extending the instrument's capabilities to nephelometric methods of analysis, including assays for specific proteins by immunoprecipitation: IgG, IgA, IgM, and transferrin. The scattered light from a green-light-emitting diode (peak wavelength 565 nm) is measured at an angle of 37.5 degrees by the existing optical detection system of the TDx analyzer without modification of the instrument. Measurements are made at quasi-equilibrium, with sample blank correction. No sample pretreatment is required, and antigen-excess is checked automatically. CVs range from 3 to 6% (within-run) and 7 to 9%. (total). Calibration curves may be stored for at least two weeks. A nephelometric method for monitoring chromogenic reactions in the green wavelength region is also described. This method--Scattered Energy Attenuation (SEA)--has been used in preliminary experiments to measure calcium, total protein, iron, and bilirubin.

Autoanalysis↗
PubMed1982

Intracellular transport of cholesterol to the plasma membrane.

We have modified a plasma membrane isolation procedure which utilizes DEAE-Sephadex beads (Gotlib, L. J., and Searls, D. B. (1980) Biochim. Biophys. Acta 602, 207-212) to rapidly measure intracellular transport of cholesterol from the site of synthesis in the endoplasmic reticulum to the plasma membrane. This transport process is rapid, with a half-time of about 10 min, has different kinetics from that of intracellular glycoprotein transport, and appears to be energy-dependent.

Acid Phosphatase↗
PubMed1980

Uptake and metabolism of fatty acids by dispersed adult rat heart myocytes. I. Kinetics of homologous fatty acids.

An adult rat heart myocyte preparation was used to study the uptake and metabolism of the 1-(14)C-labeled free fatty acids decanoate, laurate, myristate, palmitate, and oleate at 37 degrees C in the absence of serum albumin. The rate of total uptake consisted of both a nonsaturable and a saturable component. The nonsaturable component corresponded to the fatty acid accumulating in the free fatty acid fraction, and the rate of this accumulation increased logarithmically as a function of chain length. The saturable component corresponded to that portion of fatty acid converted to the only detectable metabolic products: CO2, triglyceride, and polar lipid. The Km for this process was about 1 microM and was independent of chain length. The Vmax for the saturable component varied only slightly with chain length, from 20 +/- 1 nmol/h-mg of cell protein for decanoate to 47 +/- 18 nmol/h-mg of cell protein for palmitate. The relative product distribution did vary with chain length, however, ranging from primarily carbon dioxide for decanoate to approximately equal quantities of carbon dioxide, triglyceride, and polar lipid for palmitate. Two internal pools of free fatty acid are postulated: a minor pool that equilibrates rapidly with external fatty acid and serves as the precursor for fatty acid activation, and a major pool containing most of the accumulated free acid. These two pools are interconvertible. The data support a simple diffusion or membrane-partitioning process for the accumulation of fatty acid in the second pool. The data presented in this paper are not sufficient to distinguish between a simple diffusion or a carrier-mediated process for uptake into the first pool. The saturation kinetics observed appear to represent a metabolic step such as fatty acid activation, rather than a transport carrier. Evidence of toxicity at a higher concentration of the longer chain fatty acids limits the concentration range that can be studied in the absence of albumin. Decanoate did not appear to be toxic at concentrations up to 300 microM, but laurate at 10 microM and myristate at 5 microM appeared to uncouple respiratory control.

Animals↗