Andras G. Foti

Radioimmunochemical Measurement of Bone Marrow Prostatic Acid Phosphatase

Human acid phosphatases are ubiquitous phosphohydrolases that are present in most respiring tissues and cells. Specifically, human prostatic acid phosphatase is a unique enzyme within a vast family of acid phosphatases concerned with catabolic processes in cellular metabolism. The majority of serum and bone marrow acid phosphatases are of non-prostatic origin and are present chiefly in erythrocytes, leukocytes, platelets and other maturing cells in the bone marrow. The specific concentration of prostatic acid phosphatase in serum and bone marrow is normally relatively low compared to non-prostatic acid phosphatases. Many falsely positive assays for total serum acid phosphatases and bone marrow acid phosphatases have been reported, particularly after traumatic marrow biopsy procedures and mishandling of blood samples in the clinical laboratory and in hematologic disease states. The disruption and lysis of whole blood and marrow cells can liberate non-specific acid phosphatases into the serum. Since standard enzymatic assays do not discriminate accurately prostatic acid phosphatase from non-prostatic acid phosphatase present in the serum spurious results can be realized. A preliminary experience with a promising radioimmunoassay for the specific measurement of prostatic acid phosphatase in bone marrow and serum is presented.

The effect of antibody on human prostatic acid phosphatase activity—I: Temperature and pH stabilization of acid phosphatase enzyme activity by rabbit antibody to acid phosphatase

Abstract Prostatic acid phosphatase was purified from prostatic fluid. Monospecific antisera to the purified acid phosphatase was then produced in rabbits. When antibody was coupled with acid phosphatase, the enzymatic activity was markedly stabilized against pH and temperature degradation. Both acid phosphatase and rabbit anti acid phosphatase were non specifically coupled to Sepharose-4B using cyanogen bromide. Under these circumstances slight stability occurred when antibody was bound to Sepharose, and then acid phosphatase added to the gel antibody complex. When acid phosphatase was complexed to Sepharose, no stabilization occurred.

Production of specific antibody to purified prostatic acid phosphatase

SummaryProstatic acid phosphatase may well be a prime antigenic protein in prostatic tissue and fluid. Extraction of the enzyme in highly purified form from prostatic fluid and benign hypertrophic prostatic tissue provides a unique antigen capable of inducing a prompt and specific antibody response in the goat and rabbit as manifested by immunodiffusion, immunoelectrophoresis, and immuno-fluoresence techniques. In prostatic cancer patients with elevated serum acid phosphatase levels it is possible to detect humoral circulating PAP antigen by standard immunoelectrophoretic methods and to confirm the existence of the enzyme by radioautography, L-tartrate inhibition, and the Gomori or Burstone staining procedures. Preliminary indirect prostatic immunofluorescence studies consistently demonstrated characteristic fluorescent foci in the paranuclear areas of benign prostatic epithelial cells, the presumed area of synthesis of prostatic acid phosphatase. Consideration has been given to the possibility of the development of a radioimmunoassay for prostatic acid phosphatase utilizing a heterologous antiserum to the enzyme extracted from human prostatic fluid.

The effect of antibody on human prostatic acid phosphatase. Substrate utilization by enzyme or enzyme-antibody complex.

Abstract Rabbit antibody to human prostatic acid phosphatase stabilizes the enzyme activity against thermal inactivation at antigen-antibody equivalents of two or greater. The pH optimum of the enzyme itself and the enzyme-antibody complex varies with different substrates. As the size of the substrate increases the activity of the enzyme alone decreases and antibody inhibition of enzymic activity is enhanced.

Central Noradrenergic Mechanisms in Hypertension and in Postural Hypotension

Recently, we described evidence implicating the central sympathetic nervous system in the pathogenesis of primary hypertension (1, 2). We have also reviewed reports of enhanced peripheral sympathetic tone in primary hypertension (3). Earlier, Ziegler and his colleagues (4) described low-plasma norepinephrine and a blunted norepinephrine response to postural stress in patients with orthostatic hypotension. When these patients assume an upright posture, they have a hemodynamic state that is, in some respects, opposite to that of some patients with primary hypertension (5). Herein, we report the concentrations of norepinephrine, in both plasma and spinal fluid while supine and in plasma while standing, and the magnitude of changes after standing, in patients with either primary hypertension or postural hypotension and compare them with respective values of normotensive patients.