PubMed HealthSearch

Biomedical subjects

K O Ash

Publications and source records attributed to K O Ash.

14 recordsLinked to original sources

A simple ICP-MS procedure for the determination of total mercury in whole blood and urine.

A simple and sensitive procedure for total mercury in whole blood and urine using inductively coupled plasma-mass spectrometry (ICP-MS) is described. Specimens are prepared by precipitation-extraction with 50% v/v hydrochloric acid containing EDTA and cysteine, centrifuged, and filtered through fritended screening column; the filtrates are directly analyzed by ICP-MS. The method is linear between 2 and 200 micrograms/L in the specimen with an absolute sensitivity of 0.2 microgram/L in the final supernatant. The assay variability at various concentrations (microgram/L) of mercury are as follows: intra-assay whole blood (n = 20)-4.6 +/- 0.6 (c.v. 12.3%), 18.3 +/- 1.1 (c.v. 6.1%), 56.4 +/- 2.8 (c.v. 5.0%); inter-assay whole blood (n = 15)-5.7 +/- 1.0 (c.v. 16.8%), 19.7 +/- 2.7 (c.v. 13.5%), and 50.1 +/- 6.9 (c.v. 13.7%); urine (n = 20)-9.3 +/- 1.2 (c.v. 12.9%), 29.6 +/- 2.2 (c.v. 7.4%). Recovery of organic and inorganic mercury from blood samples ranges from 91.6% to 110.2%. The method is suitable for analysis of total mercury, both organic and inorganic, in whole blood and urine.

Argon

Using 16O35Cl to correct for chloride interference improves accuracy of urine arsenic determinations by inductively coupled plasma mass spectrometry.

We have observed inaccurate urine arsenic values with the method of isobaric fractionation, which was designed to correct for the 40Ar35Cl interference with 75As quantitation by inductively coupled plasma mass spectrometry. Isobaric fractionation, which is based on ion intensities at m/z 77 and 82, consistently underestimates the 40Ar35Cl interference and overestimates urine arsenic. We present an improved method for identifying the argon-chloride interference. We observed that signal intensities for the species 16O35Cl and 40Ar35Cl are proportional (I75 = 0.0295 x I51 - 14.7, r2 = 0.998; where Ix is the normalized ion intensity at m/z X) in water and urine, over a broad range of chloride concentrations (0-800 mmol/L). The proportionality constant is remarkably stable within a run (mean and SD, 0.0295 +/- 0.0023, based on 10 replicates of five chloride calibrators, 0, 100, 200, 400, and 800 mmol/L). Increased sensitivity (50-fold) for detecting the 40Ar35Cl interference provides improved accuracy for urine arsenic quantitation as demonstrated by a split-sample comparison with graphite-furnace atomic absorption spectrophotometry.

Arsenic

Changes in sodium-lithium countertransport correlate with changes in triglyceride levels and body mass index over 2 1/2 years of follow-up in Utah.

We have previously reported from a cross-sectional study that plasma total cholesterol, triglycerides, and HDL-C were significantly and independently correlated with Na(+)-Li+ countertransport. These original participants were rescreened 2 1/2 years later (range of 20-58 months), with lipid, blood pressure, and Na(+)-Li+ countertransport measurements from both visits on 906 normotensive adults. The correlation found between age- and sex-adjusted triglyceride levels and Na(+)-Li+ countertransport at visit 1 (r = 0.34, p less than 0.0001) was reconfirmed at visit 2 (r = 0.32, p less than 0.0001). The Na(+)-Li+ countertransport correlations with HDL-C (r = -0.11, p less than 0.01) and body mass index (r = 0.24, p less than 0.0001) also remained at visit 2. After 30 months, there were significant increases of triglyceride, body mass index, blood pressure, and Na(+)-Li+ countertransport levels, and significant decreases of HDL-C and total cholesterol levels, after adjusting the changes in these variables between visit 2 and visit 1 for age, sex, time between the two visits, and the visit 1 level of the variable. Increases in triglycerides, cholesterol, and body mass index were significantly correlated with increases in Na(+)-Li+ countertransport (r = 0.23, r = 0.19, and r = 0.21, respectively). The correlations of the lipid and lipoprotein changes with Na(+)-Li+ countertransport changes were independent of body mass index and blood pressure changes. We conclude that increasing plasma triglyceride levels and body mass index are associated with increasing Na(+)-Li+ countertransport levels in both cross-sectional and longitudinal data.

Adolescent

Population-based frequency of dyslipidemia syndromes in coronary-prone families in Utah.

The frequency of familial dyslipidemia syndromes was determined from blood tests in 33 objectively ascertained families with early coronary heart disease (CHD) (two or more siblings with CHD by the age of 55 years). Three fourths of persons with early CHD in these families had 90th percentile lipid abnormalities (cholesterol level at or above the 90th percentile, triglyceride level at or above the 90th percentile, and/or high-density lipoprotein cholesterol (HDL-C) level at or less than the 10th percentile). The HDL-C and triglyceride abnormalities were twice as common as low-density lipoprotein-cholesterol abnormalities. The most common syndromes found were familial combined hyperlipidemia (36% to 48% of families with CHD), familial dyslipidemic hypertension (21% to 54% of families with CHD), and isolated low levels of HDL-C (15%), with overlapping familial dyslipidemic hypertension with familial combined hyperlipidemia and low-level HDL-C. Well-defined monogenic syndromes were uncommon: familial hypercholesterolemia being 3% and familial type III hyperlipidemia, 3%. Another 15% of families with CHD had no lipid abnormalities at the 90th percentile. Physicians should learn to recognize and treat these common familial syndromes before the onset of CHD by evaluating family history and all three standard blood lipid determinations. Failure to recognize and treat them leaves affected family members at high risk of premature CHD.

Cholesterol, HDL

Predictive value of a short dietary questionnaire for changes in serum lipids in high-risk Utah families.

Dietary questionnaires and serum cholesterol, triglycerides, and high-density lipoprotein (HDL) cholesterol determinations were completed for 1239 subjects aged greater than or equal to 20 at each of two separate screenings. The mean time between screenings was 2.5 y. After correcting for potential confounding variables, reduction of a measure of dietary cholesterol and saturated fatty acids assessed by two simple questions was a significant independent predictor of reduction in total cholesterol in serum (p less than 0.005). Initial body mass index (BMI) and change in BMI were highly significant predictors of initial values and changes in total cholesterol, triglycerides, and HDL cholesterol in serum. Reduction of dietary saturated fatty acid and cholesterol was significantly correlated with initial serum cholesterol levels, which suggest that serum cholesterol screening may be an important motivating factor for dietary change. Important public health and research implications of these findings are discussed.

Adult

Mechanism of false-negative urine cannabinoid immunoassay screens by Visine eyedrops.

To drug-free urine specimens, we added the following drugs of abuse to give concentrations twice the cutoff value for positive test results: 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (9-carboxy-THC), oxazepam, secobarbital, morphine, benzoylecgonine, amphetamine, or phencyclidine (PCP). Visine was then added. Although measured concentrations of several drugs were decreased in the presence of Visine, false-negative results were obtained only for 9-carboxy-THC for the EMIT-d.a.u. and TDx urine cannabinoid assays. Visine also decreased 9-carboxy-THC as measured by the Abuscreen assay. At low concentrations of Visine, false-negative cannabinoid results were attributable to the benzalkonium chloride ingredient of Visine. The added Visine was not detectable by routine urine analysis and had no effect on the activity of the glucose-6-phosphate dehydrogenase-drug conjugate used in the EMIT-d.a.u. assays. Moreover, analysis by gas chromatography/mass spectrometry showed no chemical modification or loss of 9-carboxy-THC in the Visine-adulterated urine specimens. However, Visine did increase the adhesion of 9-carboxy-THC to the borosilicate glass specimen containers. Results of ultrafiltration studies with Visine suggest that 9-carboxy-THC partitions between the aqueous solvent and the hydrophobic interior of benzalkonium chloride micelles, thereby reducing the availability of 9-carboxy-THC in antibody-based assays.

Amphetamine

Erythrocyte cation transport activities as a function of cell age.

Erythrocyte cation transport systems were evaluated on cell fractions from 17 subjects. Density centrifugation was used to separate washed red cells into fractions enriched with younger and older cells; the cell age differences in these fractions were verified by reticulocyte counts (means are 3.5% for younger cell fractions and 0.7% for older cell fractions). Red cell age has a pronounced effect on several cation transport activities. The older cell fractions had increases in lithium-potassium cotransport (p less than 0.001), the rate constant for the lithium-potassium cotransport (p less than 0.001) and cellular cation permeability. The older cells had decreases in the number of ouabain binding sites (p less than 0.001), the rate constant for sodium efflux via the sodium-potassium adenosine triphosphatase pumps (p less than 0.001) and the sodium-lithium countertransport (p less than 0.025). In subjects with markedly different cell ages, these effects should be considered when evaluating red cell cation transport activities.

Adolescent

Manual fluorometry of phenylalanine from blood specimens collected on filter paper: a modified procedure.

In this manual fluorometric method, blood samples are used that have been impregnated on the filter paper, a convenient collection technique that is widely used to screen newborns for phenylketonuria. The modified procedure, based on the method of McCaman and Robins [J. Lab Clin. Med. 59, 885 (1962)], includes elution of phenylalanine from specimens on filter paper, removal of proteins by precipitation with trichloroacetic acid at 0 degrees C, and then reaction with ninhydrin-peptide reagent for color development. The standard curve is linear to at least 200 mg/L and the CV is 6.3% for a phenylalanine concentration of 27 mg/L. The modified procedure is suitable both for early screening for phenylketonuria and for monitoring blood phenylalanine of phenylketonurics during dietary therapy.

Blood Specimen Collection

High-density lipoproteins estimated by an enzymatic cholesterol procedure, with a centrifugal analyzer.

We describe an assay for high-density lipoprotein cholesterol, adapted to a centrifugal analyzer, the GEMSAEC System 3, which includes use of an increased Mn2+concentration (91 mmol/liter) [J. Lipid Res. 19, 65 (1978)] and ethylenediaminetetraacetate [Clin. Chem. 22, 98 (1976)]. Modifications to the GEMSAEC system include reducing the mixing burst and preconditioning the sample tip. Accuracy of this procedure, as assessed by analysis of a control pool from the Center for Disease Control, was 99.2%. Day-to-day precision for two control pools was 320 +/- 13 and 506 +/- 17 mg/liter. Serum sample volume was decreased to 0.5 ml. In blanks with heparin/Mn2+ present, the pseudocholesterol concentrations resulting from a reaction of the enzymatic cholesterol reagent and the heparin/Mn2+ precipitating reagent depend on the source of the enzymatic reagent and appear to be enhanced slightly by the use of ethylenediaminetetraacetate. Pseudocholesterol concentrations reach a maximum at heparin/Mn2+ concentrations well below those needed to completely precipitate the low-density and very-low-density lipoprotein fractions. Population reference values were obtained from analyses done on 224 local physicians (mean: male, 500 mg/liter; female, 620 mg/liter) and 156 ambulatory patients (mean: male, 463 mg/liter; female, 553 mg/liter).

Autoanalysis

A protocol for the adoption of analytical methods in the clinical chemistry laboratory.

The demand on the clinical chemistry service laboratory to continually adopt more precise and accurate methodologies makes it mandatory for the service laboratory to have a systematic protocol for evaluating and adopting new assay methods. A protocol is presented for adopting clinical chemistry methods in the service laboratory, with emphasis on spectrophotometric determinations. The recommended protocol may be used to evaluate alternate procedures to those currently in use as well as procedures for additional analytes. We present those critical points to be considered in method adoption and instruction for the resolution of each point.

Chemistry Techniques, Analytical

Effect of evaporation from sample cups on urea nitrogen assay.

The effects of evaporation from sample cups for urea nitrogen assays were determined. Thirty replicate analyses were performed on each of five groups of pooled human serum subjected to controlled evaporation in our laboratory. We found a mean increase in urea nitrogen concentration of one percent per hour due to evaporation. We urge laboratory personnel to take the action necessary to minimize errors due to evaporation. Specimens should always be capped while awaiting analysis in the laboratory.

Blood Urea Nitrogen

Gas-chromatographic method for plasma acetate analysis in acetate-intolerance studies.

We describe a modified gas-chromatographic method for acetate in serum or plasma, intended for use in the investigation of acetate intolerance in hemodialysis patients. The assay may be adapted for use with a single-column gas chromatograph equipped with a flame ionization detector. The analysis, made isothermally, requires only 0.5 ml of plasma or serum. Only one deproteinizing step is required to prepare the sample for analysis. Additionally, we present preliminary findings of an ongoing acetate-intolerance study.

Acetates

Bilirubin-protein interactions monitored by difference spectroscopy.

Difference spectroscopy is used to monitor bilirubin-protein interactions, to assess the residual binding capacity of proteins for bilirubin. A change in the difference spectra monitored at 482 nm is directly proportional to bound bilirubin up to a molar ratio of bilirubin to albumin of approximately 1; increasing bilirubin beyond the 1:1 molar ratio does not further change the difference spectra. After excess free bilirubin is added, the change in the difference spectrum is proportional to the residual binding capacity of the serum for bilirubin. The risk of kernicterus among neonates may be assessed by monitoring the residual bilirubin binding capacity of serum. This report summarizes our research effort leading to an assay method which requires only 40 microliter of serum and can be completed in less than 10 min.

Adult

Modified micro-scale enzymatic method for plasma ammonia in newborn and pediatric patients; comparison with a modified cation-exchange procedure.

We modified two "kit" procedures for plasma ammonia to make them suitable for use with newborn and pediatric patients. Available methods either require too large a specimen, are insufficiently sensitive in the normal range, or are too cumbersome. One was an enzymatic (Sigma Chemical Co., St. Louis, Mo. 63178), the other a cation-exchange (Hyland, Costa Mesa, Calif. 92626) method. The modified micro-scale enzymatic method requires 100 microliter of plasma (CV at 38 micronmol/liter, 8%). Reagent costs per patient assay, including controls and standards, are currently $0.53. An assay run can be completed in 30 min. The modified cation-exchange method requires 200 microliter of plasma; the within-run CV is 12% for a concentration of 34.2 micronmol/liter. Reagents currently cost $0.74 per assay and each run requires less than 2 h to complete. The correlation coefficient for the two methods was 0.98.

Ammonia