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J Harter

Publications and source records attributed to J Harter.

8 recordsLinked to original sources
PubMed2006

Fully integrated whole blood testing by real-time absorption measurement on a centrifugal platform.

We present a novel microfluidic concept to enable a fast colorimetric alcohol assay from a single droplet of whole blood. The reduced turn-around time of 150 seconds is, on the one hand, achieved by a full process integration including metering, mixing with reagents, and sedimentation of cellular constituents. On the other hand, our novel total internal reflection (TIR) scheme allows to monitor the increase of the absorbance values in real-time. Thus, the saturation values can be predicted accurately based on an extrapolation of real-time measurements acquired during a 100 second initial period of rotation. Additionally, we present a metering structure to define nanolitre sample volumes at a coefficient of variation (CV) below 5%.

Blood Chemical Analysis↗
PubMed1989

Effect of L-lysine on cytosolic calcium homeostasis in cultured human normal fibroblasts.

L-lysine, a cationic essential amino acid, has been reported to affect calcium transport in both intestine and kidney. In order to investigate whether this effect is associated with changes in cytosolic calcium homeostasis, we studied the effect of L-lysine deprivation on intracellular calcium concentration ([Ca2+]i), as well as 45Ca efflux and accumulation in normal human fibroblasts. Steady state [Ca2+]i, measured using fura-2 fluorescence in cells cultured for 18 hours in a L-lysine-free medium, was significantly higher than in cells grown in the presence of as little as 4 microM L-lysine. L-lysine deprivation also led to a significant decrease of 45Ca fractional efflux compared with cells grown in complete medium. This effect was paralleled by a significant decrease in 45Ca accumulation. Lack of L-arginine from the growth medium for the same time period had no effect on either [Ca2+]i, 45Ca efflux, or 45Ca accumulation rate. Presumably, the lack of L-lysine for a significant amount of time impairs the active mechanisms of calcium extrusion, which is only partially compensated by a reduction of calcium accumulation rate. This leads to an increased steady state [Ca2+]i. It is concluded that L-lysine is an important modulator of cytosolic calcium homeostasis.

Calcium↗
PubMed1989

Enzyme product blot for nondestructive assay of protein catalytic function in polyacrylamide gels.

A new method that permits rapid, sensitive, and specific enzymatic assay of proteins in polyacrylamide gels is described. The enzyme product blot described in this report involves percolation of the reaction mixture through a gel containing native enzyme which converts the labeled substrate to a labeled product with differing chemical properties. A permeable membrane with specific ligand-binding properties overlies the gel and binds the enzyme product, but not the substrate, as reaction mixture is blotted vertically. This membrane is washed free of substrate and the location of the product is identified by autoradiography. The autoradiogram is compared with the stained gel in order to recover the enzyme for amino acid sequence analysis. The enzyme product blot is demonstrated using glycerol kinase and hexokinase.

Autoradiography↗
PubMed1989

Synthesis and characterization of a bovine hexokinase 1 cDNA probe by mixed oligonucleotide primed amplification of cDNA using high complexity primer mixtures.

Hexokinase (EC 2.7.1.1) catalyzes the first step in glucose metabolism, using ATP for the phosphorylation of glucose to glucose 6-phosphate. A portion of the HK1 gene was cloned by mixed oligonucleotide primer amplification of cDNA using primers of high complexity. The amino acid sequence for a partial fragment of bovine cardiac muscle HK was determined and used to create primer mixtures of 256- and 1024-fold complexity. Two products were generated from bovine cardiac muscle cDNA which show 82% nucleotide and 93% amino acid identity with a region of rat brain HK1 and cDNA. This work demonstrates that extension and amplification of cDNA probes may be successful even when amino acid sequence data indicate substantial codon degeneracy.

Amino Acid Sequence↗
PubMedDate not supplied

Higher corticosterone values at a fixed single timepoint in serum from mice "trained" by prior handling.

Results of "training" rodents for bioassay of synthetic corticosteroids are not consistent with expectations presumably based upon earlier implied claims stemming, perhaps, from over behavioral patterns displayed by handled as compared to nonhandled rodents (or the observer's wishful thinking?). At the timepoints investigated, "habituation" of mature mice and rats by exposure to handling, injection and killing procedures results in statistically significantly increased corticosterone concentrations. For younger animals the elevation in corticosterone concentration may also be applicable; although, it is not known to what extent we are dealing with an effect of "habituation" and/or age. Lowest hormone concentration occurred in the groups of mice housed undisturbed in an animal chamber separate from the room housing mice subjected to habituation. The results of this study suggest that for certain rodents kept in L from 0600 to 1800 "habituation" for 1 week, at least at the circadian stage corresponding to 0800, is not only ineffectual (in eliminating the rise in serum corticosterone associated with handling) but detrimental to the evaluation of synthetic corticosteroid potency by bioassay (to the extent that such a rise in exaggerated rather than reduced). In these acute experiments, methylprednisolone was more suppressive than dexamethasone and the water soluble ester-salts less suppressive than the 21-hydroxy bases. These results suggest the possibility of important differences in the distributive stage and/or the metabolism of these compounds--factors affecting the speed of onset of the desired as well as undesired effects.

Adrenal Cortex Hormones↗