Moderate Hemolysis Research Articles

Iron toxicity screening

Fischer's method for rapid detection of acute iron toxicity is modified to suit pediatric cases. TPTZ (2,4,6-tripyridyl-s-triazine) is the chromogen of choice since in a small volume of serum slight to moderate hemolysis can cause a false positive result bathophenanthroline. Ordinary labware is amenable to this simplified procedure.

Autoimmune hemolytic anemia in ulcerative colitis

Three cases of autoimmune hemolytic anemia in association with idiopathic ulcerative colitis are reported; the literature is reviewed; and therapeutic modalities are considered. Successful treatment is shown to have resulted with steroids alone, steroids and immunosuppressives, splenectomy, and colectomy. It is concluded that moderate or severe hemolysis should be treated first with high-dose corticosteriods; if unsuccessful, immunosuppressive therapy may be added or a splenectomy performed. Finally, total colectomy should be reserved for fulminant colitis and its complications and is not indicated solely for hemolysis.

Biochemical changes in a 100 km run: Proteins in serum and urine

Eleven male subjects took part in a 100 km running competition. Alterations in the total plasma protein and in ten individual plasma protein concentrations in blood and urine were measured prior to the run, immediately after and after 1 day of recovery. Five individual proteins showed a 7-10%, and lysozyme a 40%, increase in the plasma after the run. On the contrary, the haptoglobin concentration fell to 40% of its pre-race level. None of these variations were correlated with the plasma volume change. The present data showed a moderate hemolysis, as evidenced by plasma lysozyme and hemoglobin-haptoglobin binding. The urinary excretion of plasma proteins was slightly increased, especially albumin and alpha1-acid-glycoprotein. The renal clearance of plasma proteins revealed that the 100 km run induced a moderate increase of glomerular permeability without any signficant change in the tubular reabsorption process.

Fragmentation and Myelin Formation in Hereditary Xerocytosis and Other Hemolytic Anemias

Erythrocytes from a heterogeneous group of hemolytic anemias have been found to release acetylcholinesterase-enriched fragments and show myelin forms during ATP depletion in vitro. The highest amount of fragmentation was found in hereditary spherocytosis and xerocytosis, two inherited membrane defects. Our data suggest ATP depletion plays a role in producing fragmentation or myelin forms. The addition of external CaCl2 1 mM had no effect on the degree of fragmentation. However, propranolol hydrochloride, a cationic anesthetic that does not prevent ATP depletion, inhibited fragmentation and the appearance of myelin forms in both hereditary spherocytes and xerocytes. A more detailed study of the xerocyte fragments showed that they had the same protein composition as those from normal red cells, primarily integral membrane proteins and glycoproteins. The red cells from patients with PNH and G6PD deficiency had the shortest survival in vivo (51Cr) and produced the smallest amount of fragmentation and myelin forms in vitro, whereas xerocytosis with only mild to moderate hemolysis in vivo was associated with the highest amount of myelin forms and membrane fragments in vitro.

Fragmentation and myelin formation in hereditary xerocytosis and other hemolytic anemias

Erythrocytes from a heterogeneous group of hemolytic anemias have been found to release acetylcholinesterase-enriched fragments and show myelin forms during ATP depletion in vitro. The highest amount of fragmentation was found in hereditary spherocytosis and xerocytosis, two inherited membrane defects. Our data suggest ATP depletion plays a role in producing fragmentation or myelin forms. The addition of external CaCl2 1 mM had no effect on the degree of fragmentation. However, propranolol hydrochloride, a cationic anesthetic that does not prevent ATP depletion, inhibited fragmentation and the appearance of myelin forms in both hereditary spherocytes and xerocytes. A more detailed study of the xerocyte fragments showed that they had the same protein composition as those from normal red cells, primarily integral membrane proteins and glycoproteins. The red cells from patients with PNH and G6PD deficiency had the shortest survival in vivo (51Cr) and produced the smallest amount of fragmentation and myelin forms in vitro, whereas xerocytosis with only mild to moderate hemolysis in vivo was associated with the highest amount of myelin forms and membrane fragments in vitro.

Late Clinical Problems with Beall Model 103 and 104 Mitral Valve Prostheses: Hemolysis and Valve Wear

The purpose of this study was to determine the influence on hemolysis of the spatial orientation of the struts in the Beall mitral valve prosthesis, Models 103 and 104. Thirteen pairs of patients were selected to match struts oriented parallel and perpendicular to the left ventricular outflow tract axis. The average time after operation was 3.73 years. Complete blood counts and relative serum chemistry values were determined. The results demonstrated that strut orientation made no significant contribution to the mild to moderate hemolysis of normally functioning, non-worn Beall mitral valves. However, 10 of the 26 patients had marked elevations of lactic dehydrogenase (≥975 mIU/ml) and were anemic in spite of iron therapy and normal serum iron and total iron-binding capacity, in contrast to the other 16 patients. Three of these 10 patients have had their prosthetic valve removed, and all 3 valves showed moderate to severe wear of the disc and Teflon-coated struts. Six additional valves have been recovered; 3 showed moderate to severe wear and 3 had minimal wear. A marked degree of anemia, elevation of the LDH (≥975 mIU/ml), and symptoms of mitral stenosis or insufficiency in patients with the Beall prosthesis are highly suggestive of valve wear, and early operation for Beall mitral valve replacement is advocated.

Simple solid-phase radioimmunoassays for total tri-iodothyronine and thyroxine in serum, and their clinical evaluation

1. 1. Simple solid-phase radioimmunoassay methods for total tri-iodothyronine and thyroxine in serum or plasma are described. By using antibodies that are covalently coupled to microcrystalline cellulose, virtually identical assay procedures can be used for the determination of both hormones. An alkaline sodium glycinate buffer provides better assay conditions than the buffers often recommended for thyroid hormone radioimmunoassay. 2. 2. Assay results are unaffected by moderate sample haemolysis. Plasma samples stored at room temperature for more than nine days often show an apparent increase in concentration of both thyroid hormones. 3. 3. Serum tri-iodothyronine and thyroxine concentrations in healthy euthyroid subjects, and in euthyroid pregnant women are reported. In a series of 100 consecutive patients referred to a thyroid clinic the tri-iodothyronine assay discriminated better than the thyroxine assay between hyperthyroid and euthyroid patients. The thyroxine assay was much better than the tri-iodothyronine assay in discriminating between hypothyroid and euthyroid patients.

Mechanical fragility of erythrocytes in normals and in patients with heart valve prostheses.

Abstract. Hemolysis following insertion of heart ball valves shows great individual differences. Besides the mechanical properties of the prosthetic valves, individual variations in the mechanical fragility (MF) of the erythrocytes might be important in this respect. The MF has been studied with the hemoresistometer of Fleisch and Fleisch, which exposes the cells to a standardized trauma by rotation of the blood in a cylindrical chamber. The MF is defined as the amount of heme liberated. A normal material is presented, and the error of the method and the normal variation of the MF are estimated, using two different rotors. The hemoresistometer seems to be a valuable tool for evaluation of the MF, offering several advantages compared to other methods. Reference has been made to a material of nine consecutive patients with heart valve disease examined before and after prosthetic ball valve implantation. All had a normal MF before the operation, and the mean MF remained normal in spite of moderate intravascular hemolysis developing in all but one patient postoperatively. In four patients with valve prostheses readmitted for hemolytic anemia the plasma heme concentrations were markedly increased, but the MF was normal. The question whether a lowered MF may be the cause of the severe intravascular hemolysis developed by some patients remains unsettled, but the present study suggests that individual differences in MF are of minor importance for the development of severe hemolytic anemia in these patients.

The hemolytic process of viral hepatitis inchildren with normal or deficient glucose-6-phosphate dehydrogenase activity

The incidence and degree of the hemolytic process associated with viral hepatitis wasstudied in 125 children; 16 were deficient in erythrocyte G-6-PD and 5 were heterozygotes for β-thalassemia. Hemolysis was observed in 24 (23 per cent) of the nondeficient subjects, 14 (87 per cent) of those with G-6-PD deficiency, and in 4 (80 per cent) of the heterozygotes. In the nondeficient patients hemolysis was unusual and mild. Moderate and severe hemolysis occurred only in 6 children with G-6-PD deficiency and in 3 with thalassemia trait. Bilirubinemia was generally more pronounced in the patients with the erythrocyte defects; levels higher than 25 mg. per cent were noted in 4 of them. These findings suggest that either G-6-PD deficiency or thalassemia trait may modify the clinical course of viral hepatitis by favoring the induction of hemolysis and hyperbilirubinemia.

Moderate Intravascular Hemolysis and Experimental Venous Thrombosis in Rats

Summary and ConclusionsModerate intravascular hemolysis was produced in rats by intraperitoneal injections of hemolysate prepared from rats.1. An accelerated clotting time of recalcified plasma and an increased platelet adhesiveness were observed during the first ten hours after the injection.2. An increase of the fibrinogen and proaccelerin level of plasma were observed 24—48 hours after the injection.3. No increased antifibrinolytic activity was found.4. No significant increase in the incidence of thrombosis was observed during the first 24 hours after the injection whereas a high incidence of thrombosis indicating a hyperthrombotic state was observed 24—48 hours after the injection, at a time when the hemoglobinemia had disappeared. In a group of animals given a second injection of hemolysate 24 hours after the first injection a hyperthrombotic state was still present 25—48 hours after the first injection with an incidence of thrombosis equal to that noted 24—48 hours after a single injecion only.5. The present study may indicate that the hyperthrombotic state present during moderate intravascular hemolysis is associated with an increased formation of fibrinogen, factor V (proaccelerin) and probably factor VIII (AHG).

In vitro hemolysis of erythrocytes from vitamin E deficient rats and chicks.

Summary.When red blood cells of rats deficient in vitamin E are suspended in isotonic (0.9%) NaCl solution and allowed to stand overnight at room temperature, extensive spontaenous hemolysis occurs. This oxidative process is accelerated by incubating the suspensions at higher temperatures, hemolysis being nearly complete within one hour at 47° C.Hemolysis did not occur when purified N2 had been bubbled through the erythrocyte suspension in saline to remove free O2.Although dialuric acid increases the rate of hemolysis, the presence of this substance, hydrogen peroxide, or other added autoxidizable compound or oxidizing agent is not essential to the process.The incorporation of small amounts of methylene blue in the diet of vitamin E deficient rats affords marked but incomplete protection against in vitro hemolysis.Erythrocytes from tocopherol‐depleted chicks (on an encephalomalacia producing diet) manifesting severe deficiency symptoms are much more resistant to hemolysis than are those from rats. Spontaneous hemolysis of chick red cells in saline solution was not noted within 24 hours even when the cell suspensions were incubated at temperatures up to 47° C for 7 hours. The addition of dialuric acid caused faint to moderate hemolysis within 4 hours in about half of the birds studied.It is concluded that whereas the in vitro hemolysis of red cells may be used for assessing vitamin E deficiency in the rat, it is not a reliable criterion for determining a deficiency of this vitamin in the chick.

Studies of Rickettsial Toxins

Studies of Rickettsial Toxins

Characteristics of the Mammary Tumor Milk Agent in Serial Dilution and Blood Studies.

The mammary tumor milk agent has been shown to be essential for the regular development of mammary cancer in mice. It is usually transferred by nursing and is present in various tissues3-6 as well as the milk. The infectious agent has the characteristics of a virus. This report gives data on the activity of the milk agent for mammary cancer in blood plasma, blood-cell suspensions, and serial dilution of extracts made from mammary tissue. Blood was obtained from the hearts of mice known to harbor the agent. Either females of the A stock, 75 to 105 days of age, or females (A and C3H stocks and their hybrids) that had developed spontaneous mammary cancer were used. The blood was citrated to prevent coagulation but moderate hemolysis occurred. Separation of the cells and the plasma was by centrifugation (approximately 4,000 r.p.m. for 10 minutes) and the cells were washed with saline several times before they were used. The cells were suspended in saline and the plasma was injected either undiluted or diluted in saline (see table for details). As test animals mice were used that were susceptible for mammary cancer and lacking the milk agent. They were injected intra-peritoneally when from 3 to 5 weeks of age. The incidence in control mice maintained as breeding females was approximately 1%. The results for the blood studies are presented in Table I. Many of the mice in one series are living at 15 months of age. The donors of the lactating mammary tissue were AZF1 females that had had from 5 to 7 litters. The glands were macerated by means of a press, and serial dilutions were made with triple distilled water. Each animal received 1 cc of the suspension intraperitoneally and the surviving animals are 15 months of age. The preliminary results are tabulated in Table II.