Tamm-Horsfall protein coating of free cells in urine.

The origin of free cells in urine is difficult to determine in the absence of cellular casts. Using fluorescein-conjugated antibody to human Tamm-Horsfall protein (THP), it was demonstrated that some free cells in urine are coated with THP. To evaluate the usefulness of this observation in the differentiation of upper from lower urinary tract disease, the presence of THP coating of cells was correlated with the clinical diagnosis in 141 subjects. The percentage of THP-coated cells for each group (mean +/- SD) was Healthy volunteers (n = 10), 4% +/- 2%; hospitalized controls (n = 20), 3% +/- 3%; glomerulonephritis (n = 21), 61% +/- 6%; chronic interstitial nephritis (n = 26), 56% +/- 5%; other renal parenchymal diseases (n = 14), 50% +/- 8%; bladder disease (n = 14), 8% +/- 2%; and hypertension (n = 36), 24% +/- 33%. Based on the results from the bladder disease group, 12% coating was set as the 95% confidence limit for lower urinary tract disease. The results in this group were not different from control subjects. By analysis of variance and chi 2 analysis, subjects with renal parenchymal disease could be distinguished from those with hypertension and bladder disease (P less than .001 and P less than .0001, respectively). The presence of cellular coating by THP in renal parenchymal disease and its absence in bladder disease suggests that this simple test may be of use in determining the origin of free cells observed in the routine microscopic urinalysis.

"The Yellow IRIS" urinalysis workstation--the first commercial application of "automated intelligent microscopy".

Since 1982, "automated intelligence microscopy" (AIM) has been refined and adapted to perform the portion of the urinalysis profile traditionally done by a microscope. AIM and analytical subsystems measuring relative density and performing dipstick chemistry compose the main elements of "The Yellow IRIS" urinalysis workstation, an attended system for automation and standardization of routine urinalysis. Performance data gathered at three laboratory test sites show AIM to be analytically consistent over the required range of particulate enumeration, and show that it detects 20% more abnormalities than by conventional microscopy, and with greater precision (CVs 5 to 20%). Complete urinalysis, including the microscopic examination, requires little more than 1 min for normal specimens, 3 min for most abnormal specimens. Actual throughput rate varies with the particulate composition of specimens; typically, it averages greater than 30 specimens per hour, a 300% improvement in urinalysis productivity by CAP standards and an almost 500% improvement when typical emergency-use demands are taken into account.

Automated intelligent microscopy (AIM) and its potential application in the clinical laboratory.

The microscope is the most ubiquitous instrument in the clinical laboratory. We discuss improvements in its use, in terms of "front-end" automation of specimen handling and "back-end" automation of image analysis: automated intelligent microscopy. Examples of spatial and spectral differentiation illustrate the potential of this automated version of microscopy as a useful tool with very powerful analytical capabilities for the clinical laboratory.

Combined blood cell counting and classification with fluorochrome stains and flow instrumentation.

A multiparameter flow cytophotometer was used to count and classify fixed human blood cells fluorochromed with a mixture of ethidium bromide (EB), brilliant sulfaflavine and a blue fluorescent stilbene disulfonic acid derivative (LN). The system measures light scattered by the cells and absorption at 420 nm for all cells. In addition, nuclear EB fluorescence (540 leads to 610 nm) and cytoplasmic fluorescence from LN (366 leads to 470 nm), brilliant sulfaflavine (420 leads to 520 nm) and EB exicted by energy transfer from LN (366 leads to 610 nm) are measured for all nucleated cells. This information is sufficient to perform red and white blood cell counts and to classify leukocytes as lymphocytes, monocytes, basophils, eosinophils or neutrophils. Light scattering and/or nuclear and cytoplasmic fluorescence values may be further analyzed to obtain the ratio of immature to mature neutrophils. Counts produced by the system are in reasonable agreement with those obtained by electronic cells counting and examination of Wright's-stained blood smears; some discrepancies appear to be due to systematic errors in the manual counting method.

Norepinephrine turnover in the heart and spleen of the cardiomyopathic Syrian hamster.

Although a reduction in myocardial norepinephrine stores in cardiac hypertrophy and congestive failure is well documented, norepinephrine turnover has been inadequately studied in such hearts. We compared norepinephrine turnover in control and cardiomyopathic hamsters by following the decline in specific activity of myocardial norepinephrine after labelling with an intraperitoneal tracer dose of 3H-norepinephrine. Adult myocardial norepinephrine concentrations were not attained until 4 weeks of age in both strains. There was no difference in the rate of constant (K) for myocardial norepinephrine turnover (0.107+/-0.004 hours-1 vs. 0.100+/-0.005 hours-1) in the two strains of hamsters during the neonatal period. In young control hamsters, K fell to 0.064+/-0.004 hours-1, but that for age-matched hamsters with mild cardiac hypertrophy was 0.102+/-0.001 hours-1 (P less than 0.001). There was little change in K as control hamsters aged. With the development of more severe hypertrophy in cardiomyopathic hamsters, cardiac norepinephrine decreased and resting K rapidly increased to approach the value obtained when hamsters were subjected to immobilization stress (0.302+/-0.013 hours-1). The maximum achievable K remained the same for both control and dystrophic hamsters even during terminal disease. Prolonged immobilization led to a reduction in cardiac norepinephrine in both strains. Ganglionic blockade of failing hamsters completely restored the levels of both cardiac norepinephrine and K to control values. Splenic noradrenergic nerves showed no change in K, norepinephrine content, or maximum K during cardiac decompensation. We conclude that, in the late stages of hamster cardiomyopathy, there is a progressive and possibly specific increase in cardiac sympathetic tone which leads to a concomitant decrease in cardiac norepinephrine. With the loss of sympathetic reserve, congestive failure supervenes.

Electrocardiographic recordings for assessing survival of cardiac allografts in the hamster cheek pouch.

Techniques for the recording of electrical events associated with contraction of allografted myocardium in the cheek pouch of inbred hamsters have been developed. The results of such recordings have been useful in assessing contractility and viability of the graft. The degree of tolerance to cardiac allografts in cheek pouches of inbred hamsters appears to be a function of histocompatibility and not "immunological privilege" of the cheek pouch.