William B. Castle

Studies on the Destruction of Red Blood Cells

B OORMAN, Dodd, and Loutitl have demonstrated that the immune serum prepared by Coombs, Mourant, and Race2 by the injection of human serum (not red cells) into rabbits may be used to distinguish c’ertain types of acquired hemolytic jaundice from congenital hemolytic jaundice: They observed that the washed red blood cells from five patients with acquired hemolytic jaundice were agglutinated by this serum. On the other hand, the washed red blood cells from seventeen patients with congenital hemolytic jaundice were not agglutinated. The test was originally developed by Coombs and associates3 for the detection of the incomplete Rh agglutinin on the supposition that such sensitized red blood cells might carry adsorbed antibody globulins. The red blood cells are agglutinated presumably as a physical manifestation of the union of the globulin OP other substance on the surface of the red cells with the antibody in the test serum. Acquired hemolytic jaundice has been reported as Fan independent acute or chronic entity*> 5 or in association with malaria, tuberculosis, cirrhosis of the liver, Hodgkin’s disease, leucemia, carcinomatosis, and cysts and tumors of the ovary.sp 7 Splenomegaly is often present in such conditions, and splenectomy in some instanbes, although not necessarily altering the cause of the disease, may be beneficial by decreasing red blood cell destruction. 1:n some cases the agglutination titer of the patient’s red blood cells in antihuman serum rabbit serum declines after splenectomy.* Such observations, also encountered in our own experience, have raised the question of whether the spleen, whkh contains both lymphocytes and reticula-endothelial cells as potential sources of antibodies, might be a source of the substance presumably adhering to the red blood cells of these pa.tients. The principal objective of the experiments reported here was to examine the spleens removed from patients with acquired hemolytic jaundiee and various other conditions for the presence of substances possessing an affinity for normal red blood cells.

Erythropoietic effect of cobalt in patients with or without anemia.

COBALT in the form of one of its salts, usually cobaltous chloride or cobaltous nitrate, has been shown to produce polycythemia in amphibia, birds and mammals.1 2 3 In mammals the increased concent...

OBSERVATIONS ON THE ETIOLOGIC RELATIONSHIP OF ACHYLIA GASTRICA TO PERNICIOUS ANEMIA.: IV. A BIOLOGIC ASSAY OF THE GASTRIC SECRETION OF PATIENTS WITH PERNICIOUS ANEMIA HAVING FREE HYDROCHLORIC ACID AND THAT OF PATIENTS WITHOUT ANEMIA OR WITH HYPOCHROMIC ANEMIA HAVING NO FREE HYDROCHLORIC ACID, AND OF THE RÔLE OF INTESTINAL IMPERMEABILITY TO HEMATO-POIETIC SUBSTANCES IN PERNICIOUS ANEMIA

Este artículo clásico hace referencia a la X observación, dentro de una serie de experimentos realizados por William Bosworth Castle (fig. 1). Castle, siendo joven estudiante de Medicina de la Universidad de Harvard, y posteriormente alumno interno del departamento de Medicina Interna del Boston City Hospital, elucida a través de sus observaciones y a lo largo de su trayectoria profesional, la patogénesis de la anemia perniciosa. Hasta entonces se sabía que la anemia que aparecía en el curso de una gastritis atrófica era conocida como anemia perniciosa, denominación que fue hecha por Addison en 1855 y posteriormente corroborada por Biermer en 1872. En aquella época esta enfermedad evidenciaba un mal pronóstico pero todavía no se conocía su causa. El concepto clásico descrito por Addison de anemia perniciosa era:

Oxidant injury of caucasian glucose-6-phosphate dehydrogenase—deficient red blood cells by phagocytosing leukocytes during infection

Patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency of red blood cells (RBC) may develop sudden hemolytic anemia during infection. Since phagocytizing polymorphonuclear leukocytes (PMN) are known to generate hydrogen peroxide, we explored the influence of this oxidant product of PMN on juxtaposed G6PD-deficient and normal RBC. The oxidant stress induced by phagocytosis depleted G6PD-deficient RBC of reduced glutathione (GSH) and this was associated with rapid removal of these cells from the circulation by the liver and spleen. No such effect was observed on normal RBC. Phagocytizing chronic granulomatous disease (CGD) PMN which lack hydrogen peroxide generation, failed to diminish GSH level in G6PD-deficient RBC. Thus, PMN can pose as a source of oxidant damage to G6PD-deficient RBC due to hydrogen peroxide generated during phagocytosis.

Observations on the etiology and treatment of anemia associated with hookworm infection in Puerto Rico.

Studies were made of 83 patients in Puerto Rico who had both anemia and hookworm infection. These patients were selected primarily because of the severity of their anemia and the absence of complicating infection. The general study plan was to observe the effect of removal of the parasites of the administration of a high protein diet of various extracts of liver and of iron salts. In attempting to ascertain the etiology of anemia there were morphological considerations (blood findings bone marrow) and physiological considerations (relation of hookworm to anemia relation of other factors to anemia). Other factors examined were dietary defects and the relation of gastrointestinal pathology to the anemia. There is focus on various means of treating the anemia. 1 method of treatment involves removal of the hookworms. While other studies showed this method to be quite effective the present observations showed that the effect of removal without other therapy was minimal. Rapid improvement however was observable with the administration of iron. In fact the daily administration of 6 gm ferric ammonium citrate without parasite removal brought rapid relief to the anemia. Thus the removal of the hookworm burden has no special effect on the hematopoiesis whereas iron salt administration brings rapid hemoglobin regeneration. Various extracts of liver were found to be relatively effective. Improvement of diet accomplished by adding 1500 cc of milk and 300 gm of lean beef had no significant effect on reticulocyte production. Various study results confirmed that even under controlled hospital conditions removal of the parasite alone resulted in little improvement. Improvement in diet was unable to significantly influence the rate of hemoglobin formation. It is clear that by direct treatment of the anemia (i.e. administration of iron salts) that clinical improvement can be greatly expedited.

Mechanism of Intrinsic Factor Action in the Gastrectomized Rat.

Summary and Conclusion 1. In experiments with gastrectomized rats: (a) The probable species specificity of rat intrinsic factor was demonstrated by the failure of a highly purified preparation of hog intrinsic factor to increase the absorption of Co60-B12. (b) In consonance with previous observations in pernicious anemia, acidification (pH 1.85) of a mixture of Co60-B12 and homogenized rat stomach inhibited the absorption of Co60-B12. (c) In contrast to previous observations in pernicious anemia, separate serial administration of Co60-B12 and rat gastric juice failed to show that rat intrinsic factor was more effective in promoting the absorption of Co60-B12 when it preceded than when it followed the Co60-B12. Indeed, no certain effect of rat intrinsic factor was detected whether it preceded or followed the Co60-B12 by as short an interval as 30 minutes. 2. In experiments with paired isolated loops of small bowel with intact blood supply created in previously gastrectomized rats: (a) The radioactivity of the wall of the loop perfused with Co60-B12 and rat gastric juice was greater than that of the paired loop perfused with Co60-B12 and rat gastric juice previously heated to destroy its intrinsic factor activity. No consistent difference was observed with heated or unheated human gastric juice. (b) The absorption of Co60-B12 was greater when labelled rather than unlabelled vit. B12 in equal amounts had saturated the binding capacity of rat gastric juice before admixture with an equal amount of the other form of vit. B12 just before the intestinal perfusion. 3. In conclusion, binding of vit. B12 is an important, perhaps indispensable, aspect of the function of intrinsic factor.

Oxygen saturation of sternal marrow blood with special reference to pathogenesis of polycythemia vera.

Summary An attempt was made to measure the stimulus to erythropoiesis in the human bone marrow by estimating the percentage oxygen saturation of blood removed from the sternal marrow cavity through a needle inserted through the cortex as in the performance of a sternal needle biopsy. By this means, blood samples were collected without effective contact with air and their oxygen content, capacity and in some instances pH were determined. In general no significant differences were demonstrated between normals, convalescent controls, anemic patients, and patients with polycythemia vera. In patients with secondary polycythemia, the percentage oxygen saturation of the blood removed from the bone marrow was relatively reduced, probably entirely as a result of the manifest unsaturation of the arterial blood. In some patients with leukemia and with myeloid metaplasia and in some patients with polycythemia vera with evidence of excessive myeloid activity, the data suggest an increased local oxygen utilization relative to the blood flow in the bone marrow. However, it was concluded that the technics used were not adequate to demonstrate an anoxic stimulus to increased erythropoiesis even in the marrow of patients with chronic anemia, possibly because of the difficulty of obtaining blood samples satisfactorily representative of the undisturbed environment of the erythropoietic cells.

The Site of Vitamin B12-Intrinsic Factor Complex “Releasing Factor” Activity in the Rat Small Intestine

Summary Vitamin B12 bound to rat intrinsic factor concentrate was made dialyzable by the supernate of the homogenate of only the proximal 10% of the rat small intestine. Since the major site of absorption of vit. B12 in the rat is the midileum, the present finding throws doubt upon, but does not exclude, the possible physiologic role of such “releasing factor” in absorption of vit. B12 by the rat.

Osmotic and mechanical fragilities of erythrocytes of monkeys infected with P. knowlesi malaria.

Summary Experiments with the blood of control and of Macaca mulatta monkeys infected with Plasmodium knowlesi indicate that the parasitized red blood cells develop increased osmotic fragility and in vitro are selectively destroyed by mechanical trauma. Consequently, it is suggested that because of this increased mechanical fragility of the red blood cells the motion of the circulation in vivo plays a part in the production of the hemolytic anemia in malaria of monkeys and, by analogy, of man.

Nutritional Macrocytic Anemia: Response to a Substance Other Than the Anti-Pernicious Anemia Principle.

Recent publications1-7 indicate a difference between the basic nutritional deficiency in Addisonian pernicious anemia and in certain macrocytic anemias usually associated with pregnancy and encountered in both the tropics and the temperate zone.