Owen, Charles A.

SUMMARY Normative values were established for results of tests of blood coagulation and platelet function in swine. In swine that have von Willebrand's disease, bleeding time was prolonged, platelet retention was decreased glass bead column), and factor VIII concentration was low.

SUMMARY Factor VIII concentrations in plasma, platelet retention, and ear bleeding times were determined on 35 normal pigs, 40 obligate carriers of porcine von Willebrand's disease, and 25 bleeders. The bleeders could be distinguished from the normal pigs by each of the 3 tests. At most, only one-third of the carriers could be distinguished from the normal pigs by any one test. A hemostatic score, derived from the results of the 3 tests, increased the discrimination to slightly more than half. The degrees of abnormality on the 3 diagnostic tests seemed to vary from family to family and might have been related to the sire of the bleeder.

SUMMARY Copper (Cu) was assayed in serial liver biopsy specimens of 5 pups resulting from a mating of 2 Bedlington Terrier carriers of inherited Cu toxicosis; the latter were sibling offspring of an affected Bedlington Terrier and a normal dog. Between 5 and 7 months of age, 1 of the pups had acceptable hepatic Cu values in each of 6 specimens. The hepatic concentration of Cu in another pup increased steadily from 801 to 3,874 μg/g dry weight. The other 3 pups may be heterozygotes (carriers); in 1—the hepatic Cu peaked at 1,043 μg/g at 9 months, in the 2nd—at 636 μg/g at 7 months, and in the 3rd—at 492 μg/g at 7 months. Acceptable concentrations were present in these 3 dogs at 9 to 14 months. Results in the present study indicate that heterozygotes may have 1 normal and 1 abnormal controller gene for regulating their hepatic Cu metabolism, thus accounting for the increased hepatic Cu concentration only in the early months of life. The affected Bedlington Terrier with 2 abnormal genes was unable to suppress the progressive hepatic accumulation of Cu. Paired liver biopsy specimens obtained at 5 to 7 months and at 14 or 15 months of age might distinguish the heterozygote from the normal and the affected Bedlington Terrier pups.

A case is described in which oxalosis occurred in an 11-year-old boy. The patient had experienced the formation of numerous calculi and multiple operations for removal of the calculi. The clinical, pathologic and histochemical findings in the case are presented. The unusual features of the case are extensive deposits of calcium oxalate crystals in the kidneys, bone marrow, spleen, liver, blood vessels, and adrenal cortex. Calcium oxalate, a relatively inert substance, appears to effect renal damage by virtue of its deposition in the renal tissue. While many aspects of oxalate metabolism remain obscure, it is generally presumed that oxalosis represents an as yet undefined metabolic defect.

Abstract In 12 normal subjects, DNA and RNA synthesis by individual marrow cells was studied in vitro by autoradiography after short-term incubation with tritiated nucleosides. The DNA labeling index and RNA synthesis rate were highest in normal immature cells. The DNA index was zero in all mature cells, including small lymphocytes and plasma cells. In the most actively proliferating normal immature cells, the RNA synthesis rate was about 5 times higher than in normal plasma cells. Since both normal immature cells and plasma cells are characterized by a high content of RNA and active protein synthesis, the results of the present study suggest that a high RNA turnover may not be necessary for protein synthesis to proceed in nonproliferating, protein-producing marrow cells. In 10 patients with myeloma and in two patients with primary macroglobulinemia, the abnormal plasma cells were found to be labeled with H3thymidine in each instance. The RNA synthesis rate was about 3 times greater in plasmocytoid myeloma cells than in mature plasma cells of normal marrows, and also about 3 times greater in lymphocytoid myeloma cells than in mature lymphocytes of normal marrows. The abnormal cells in the lymphoplasmocytic disorders apparently differ from their counterparts in normal marrows not only by a higher proliferative potential, but also by a much greater rate of RNA synthesis. In actively dividing normal bone marrow cells, high values for both DNA and RNA synthesis were found. By contrast, in the neoplastic lymphoplasma cells studied, a discrepancy was noted between relatively low values of DNA synthesis and high values of RNA synthesis. Production of abnormal proteins within long-lived neoplastic plasmocytic cells may account for this finding.

Abstract When oxalated or resin-decalcified human plasma was adsorbed with barium sulfate, equilibrated with 0.0175M phosphate buffer (pH 6.3), and chromatographed through diethylaminoethyl (DEAE) cellulose, most of the plasma’s fibrinogen emerged from the column and factor XII was detected. No factor VIII was found. A second buffer phase, in which 0.04M phosphate buffer (pH 5.9) was used, removed more protein but not fibrinogen or factor VIII. A third phase, 0.10M phosphate buffer (pH 5.6), produced additional fibrinogen and, unlike the first fibrinogen fraction, factor XIII; but again, no factor VIII. A fourth and final phase, 0.40 to 0.50M phosphate buffer (pH 5.3), removed factors VIII and V, and the eluate contained no fibrinogen. Although separation of fibrinogen into two components required the four-buffer system, maximal yields of factor VIII were achieved by compressing the first three buffer phases into one. Specific activities of about 50-fold, from large volumes of plasma, and 200-fold, from small volumes, were obtained. Refined factor VIII was unstable unless citrate was added to a concentration of 0.4 per cent.

Abstract Rapid formation of C14-bilirubin from glycine-2-C14 or δ-aminolevulinic acid-4-C14 has been demonstrated in isolated, perfused rat liver. The kinetics of labeled bilirubin production in vitro were similar to those of the "early labeling" pigment fraction observed in intact rats. The magnitude of bile pigment formation from C14-ALA by isolated liver was comparable to that in intact animals. Derivation of the C14-bilirubin from hepatic degradation of labeled hemoglobin was excluded by perfusion of the liver with blood poor in reticulocytes, or with plasma. The findings identify the liver as an important source of the "early-labeling" pigment fraction.

Abstract A simple test is proposed for evaluating the gastrointestinal absorption of iron. It begins with an oral dose of 50 µg. of stable iron labeled with 1 µc. of Fe59, to which about 300 mg. of ascorbic acid are added. Feces are assayed for Fe59 until any single stool contains less than 1 per cent of the dose. Absorption is determined on the assumption that all unrecovered Fe59 has been absorbed. The test was designed to minimize handling of stools by assaying whole specimens, and this process has the added virtue of increasing statistical accuracy of the measurement of radioactivity. The main advantage of the procedure using a 50 µg. dose is that with it one obtains a normal value for absorption which is about one-half of the dose. This permits measurement of either increased or decreased iron absorption. With our standard test method, 19 normal women absorbed an average of 62 per cent of the dose and 22 normal men absorbed 40 per cent. In addition to this sex difference, not explained by the difference in their blood hemoglobin levels, there seemed to be a tendency toward reduced absorption with greater age. Preliminary results of this test in patients with hypochromic anemia and steatorrhea are reported. Increased absorption of iron followed the administration of ascorbic acid whether the iron was in the ferric or the ferrous form. The concept that ferrous iron is better absorbed by man requires re-evaluation.

Abstract Thirty-six patients with active rheumatoid arthritis in whom anemia was slight were investigated, in part with radioiron and radiochromium, for abnormalities which might lead to anemia. The primary defect appeared to be a pronounced reduction of ability to absorb iron from the intestinal tract. There was an equally marked reduction in the serum concentration of iron which could not be attributed to the slight reduction in the iron-binding capacity of the serum. Despite the reduction level of iron in the serum, the apparent turnover of iron was so rapid that the total clearance per day was normal or moderately elevated. However, the fraction of the iron cleared from the blood and appearing in the circulation in newly synthesized erythrocytes was slighfly depressed. Erythrocyte survival was normal or very slightly shortened; hemolysis seems to play no important role. The fecal excretion of radioiron, not associated with erythrocytes, was greatly increased in two patients in whom there was no evidence of intestinal bleeding. A moderate increase in plasma vohime was common.