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Biomedical subjects

D C Cannon

Publications and source records attributed to D C Cannon.

8 recordsLinked to original sources

Radioimmunoassay for immunologlobulin G in serum and urine.

We describe a radioimmunoassay for immunoglobulin G (IgG) in serum and urine. Aliquots of diluted samples and 125I-labeled IgG were incubated in antibody-coated tubes at 37 degrees C for 24 h, the supernates were decanted, and the radioactivity in tubes containing the bound fraction was counted. The dose-response curve in the range of 0.4--500 mg/L of urine or 640--40 000 mg/L of serum was linear on logit-log transformation and iterative weighted regression. Assay sensitivity was 10 ng of IgG. Validation studies included testing for precision, accuracy, antibody specificity, and parallelism of the dose-response curves for standard and unknown. In a study of 14 apparently normal individuals, serum IgG = 4.0-10.9 gL, urine IgG = 1.1-4.8 mg/24 h, and IgG clearance = 0.2 X 10(-4) to 4.8 X 10(-4) mL/min. In 20 patients with renal allografts, serum IgG = 15.8-66 g/L, urine IgG - 9.6-626 mg/24 h, and IgG clearance = 9 X 10(-4) to 1.99 X 10(-1) mL/min. IgG values correlated well with severity of renal allograft rejection.

Graft Rejection

Decreased angiotensin-converting enzyme in the adult respiratory distress syndrome.

Using hippuryl-L-histidyl-L-leucine as substrate, serum angiotensin-converting enzyme was measured in 13 patients who had adult respiratory distress syndrome, eight patients with respiratory failure without adult respiratory distress syndrome, and two groups of controls: 24 healthy blood donors and 24 hospitalized patients with a variety of conditions but without respiratory failure or adult respiratory distress syndrome. Serum angiotensin-converting enzyme expressed in units/ml was 14.60 +/- 5.60 for adult respiratory distress syndrome compared with 28.92 +/- 6.60 for the blood donors, 20.76 +/- 5.87 for the patients with respiratory failure without adult respiratory distress syndrome and 20.20 +/- 5.94 in the hospitalized patients without respiratory failure or adult respiratory distress syndrome. These differences were significant, P less than .001 when adult respiratory distress syndrome was tested against the blood donors and P less than .01 against the other two groups. The significance of these findings is not clear, but the possibility is raised that the decrease of angiotensin-converting enzyme in adult respiratory distress syndrome results from a loss of pulmonary endothelial cells, which are known both to produce angiotensin-converting enzyme and to be damaged in adult respiratory distress syndrome.

Adolescent

Radioimmunoassay for urinary albumin.

We describe a rapid, sensitive, and precise radioimmunoassay for urinary albumin (Ualb). Aliquots of diluted urine were incubated at room temperature for 1 h with 125I-labeled albumin and a rabbit antiserum monospecific for human albumin. Phase separation was effected by the double-antibody technique. The dose-response curve as linear in the range of 15.6-10000 ng, equivalent to 4 to 3000 mg/liter of urine. The limit of sensitivity was 16 ng of albumin. The coefficient of assay variation was 4.8%, both at 44 mg/liter and at 1304 mg/liter. A displacement curve obtained with a serially diluted urine sample of high albumin concentration was completely superimposable with the curve for which human albumin was used as a standard. In 26 normal individuals the range for Ualb was 2.2--12.6 mg/24h, and for albumin clearance (Calb, 1.8 x 10(-5)-19.6 x 10(-5) ml/min. After renal homografts in 25 patients, Ualb ranged from 16.9 to 9928 mg/24 h, and Calb from 2.7 x 10(-4) to 1.7 x 10(-1) ml/min. Both increased Ualb and Calb correlated well with the severity of renal homograft rejection.

Albuminuria

Spectrofluorometry of free and protein-bound plasma tryptophan.

We report a rapid, simple, and sensitive spectrofluorometric procedure for total serum or plasma tryptophan. Plasma deproteinization, tryptophan extraction, and pH optimization are all carried out with a buffered cellulose/ethanol reagent. The fluorescence is measured at an emission wavelength of 350 nm, on excitation of 290 nm. Tryptophan fluorescence relative to concentration is linear up to 50 mg/liter. The high precision, evidenced by a between-run coefficient of variation of 3.1%, negates the need for duplicate analysis of samples. Analytical recovery of tryptophan from plasma up to 50 mg/liter is 90-100%. We observed no significant difference between values for tryptophan in serum and plasma from the same individual. Assay of up to 32 samples requires 60 min. Commonly used anti-depressants have no detectable effect on tryptophan determination by this procedure.

Humans

A quality control program for the radioimmunoassay laboratory.

The major problems in applying quality control to radioimmunoassay measurements are (a) nonlinearity of the dose-response curve, and (b) nonuniformity of the residual variance. A logit-log transformation of the dose-response variables combined with an iterative weighted regression analysis appears to overcome most of the difficulties. This technic is particularly helpful when applied to substandard runs where extraneous standard points tend to distort assay results. The authors describe a quality control program that involves recording control values on calendar and histogram formats, monitoring assay variables by charting, and comparing the computer-calculated slope with the graphic plot to reveal "outliers." This program is useful in guiding technologists to locate possible causes for "out-of-limits" runs.

Computers

Determination of ionized calcium in serum that has been exposed to air.

We examined changes in ionized calcium concentration in serum after its exposure to air. Samples with total protein concentrations ranging from 50 to 90 g/liter were equilibrated with CO2 in nitrogen (5/95, by vol) or CO2 alone, to produce pH values of 7.0 to 8.0. Ionized calcium was then measured with an Orion flow-through electrode system. Curves relating pH and ionized calcium concentration had statistically identical slopes regardless of protein concentration. A factor was derived, based on pH change, for correcting values for ionized calcium in serum exposed to air, and its validity was confirmed by comparing corrected values for samples allowed to stand at ambient temperature (23 degrees C) without anaerobic precautions with values initially obtained on anaerobic aliquots of the same samples.

Air

Improved method for euglobulin clot lysis time.

1. The use of Evans Blue dye to facilitate endpoint determination the elimination of 4 degrees, assay conditions are technical improvements in the euglobulin clot lysis test. 2. Plasma samples have limited stability at 30 degrees or 4 degrees, but are stable for prolonged periods at minus 20 degrees. Samples with accelerated clot lysis are much less stable than normal samples at 30 degrees. 3. The normal range is determined as greater than 70 minutes for citrated plasma and greater than 50 minutes for oxalated plasma. There is no sex difference in the normal range.

Blood Coagulation Tests