scholarly journals Studies on Spontaneous In Vitro Autohemolysis in Hemolytic Disorders

Blood ◽  
1956 ◽  
Vol 11 (11) ◽  
pp. 977-997 ◽  
Author(s):  
LAWRENCE E. YOUNG ◽  
MARY JANE IZZO ◽  
KURT I. ALTMAN ◽  
SCOTT N. SWISHER
Keyword(s):  
Lactic Acid ◽  
Hereditary Spherocytosis ◽  
Substantial Reduction ◽  
Osmotic Fragility ◽  
Red Cells ◽  
Hemolytic Disease ◽  
Myeloid Metaplasia ◽  
Effect Of Glucose

Abstract Measurements of spontaneous lysis (autohemolysis) of red cells in sterile defibrinated blood after 48 or more hours of incubation at body temperature were found useful in the investigation of certain hemolytic states. Abnormally rapid autohemolysis was demonstrated most consistently in hereditary spherocytosis, but was also found in other types of spherocytosis and in several examples of non-spherocytic hemolytic anemia. Autohemolysis above the normal range can be regarded as strong presumptive evidence of a hemolytic disorder, but a normal rate of autohemolysis by no means excludes the possibility of a hemolytic process in any given case. Abnormalities of hereditary spherocytes causing increased autohemolysis were shown to be correlated with those responsible for spherocytosis as reflected in osmotic fragility tests. There appeared to be closer correlation with abnormalities causing increased osmotic fragility of the cells after 24 hours incubation. The autohemolysis test and the osmotic fragility test on incubated red cells were found to be equally sensitive in their capacity to detect spherocytosis of the hereditary type. Addition of adenosine, guanosine or inosine caused moderate to marked reduction in autohemolysis of nearly all types of red cells tested. Lysis of incubated spherocytes from active cases of autoimmune hemolytic disease was more markedly inhibited by a small amount of adenosine than was the lysis of hereditary spherocytes. Glucose regularly caused marked inhibition of autohemolysis of hereditary spherocytes and of red cells from patients with acute leukemia. The effect of glucose on HS red cells might have been due in part to a lowering of pH by formation of lactic acid since acidification of the blood with lactic, citric or hydrochloric acid also caused substantial reduction in autohemolysis. Glucose caused slight to marked increase in lysis of red cells in certain other cases of spherocytosis, notably in autoimmune hemolytic disease and in myeloid metaplasia. Addition of lactic acid to the blood from several of these patients had similar effect. In 2 atypical cases of chronic spherocytosis, with hematologically normal relatives, autohemolysis was accelerated by addition of glucose to the blood samples. Adenosine had little effect until after splenectomy when both glucose and adenosine inhibited hemolysis, much the same as in blood from typical cases of hereditary spherocytosis. It seems likely that when the abnormalities of carbohydrate metabolism of the red cells in certain hemolytic disorders are better understood, measurements of autohemolysis may be modified so as to enhance their usefulness in detecting and differentiating the various hemolytic states. It also seems likely that further studies on in vitro autohemolysis will help to elucidate some hemolytic mechanisms operating in vivo.

scholarly journals The Removal of in Vitro Damaged Erythrocytes from the Circulation of Normal and Splenectomized Rats

Blood ◽  
1965 ◽  
Vol 26 (1) ◽  
pp. 49-62 ◽  
Author(s):  
JOHN E. ULTMANN ◽  
CLARA S. GORDON
Keyword(s):  
Life Span ◽  
Osmotic Fragility ◽  
Reticuloendothelial System ◽  
Red Cells ◽  
In Vitro Tests ◽  
Red Cell ◽  
Cell Integrity ◽  
Prolonged Incubation

Abstract The in vitro alterations and in vivo fate of erythrocytes treated with N-ethylmaleimide or subjected to prolonged incubation were studied in normal and splenectomized rats. Minimal injury (20 µM NEM/ml. RBC) resulted in red cells with decreased osmotic fragility and increased plasticity; however, mechanical fragility was significantly increased. These cells were removed with a half-time of 78 minutes, mainly by splenic sequestration, and splenectomy prolonged their life span. Incubation at 37 C. for 21 hours produced erythrocytes with increased osmotic and mechanical fragility and decreased plasticity. Erythrocyte clearance was more rapid (T½ = 59 minutes), with spleen and liver removing approximately an equal number of cells and splenectomy having little effect on red cell life span. With severe injury (40 µM NEM/ml. RBC), all three in vitro measurements showed marked alterations, red cell removal was rapid (T½ = 35 minutes), mainly by hepatic sequestration, and clearance was unaffected by splenectomy. The present studies have shown that chemical injury or prolonged incubation lead to profound changes in in vitro tests of red cell integrity, the mechanical fragility predicting most closely the subsequent in vivo events. Although the entire reticuloendothelial system appears to participate in red cell removal, the spleen and liver are the major sites of sequestration in the rat. The splenic removal predominates with minimally injured cells, hepatic removal with moderately and severely altered cells. The type of injury appears to be of less significance than the degree of injury of the red cells.

scholarly journals HEREDITARY NONSPHEROCYTIC HEMOLYTIC ANEMIA

Blood ◽  
1950 ◽  
Vol 5 (3) ◽  
pp. 233-253 ◽  
Author(s):  
WILLIAM H. CROSBY
Keyword(s):  
Hemolytic Anemia ◽  
Hereditary Spherocytosis ◽  
Red Cells ◽  
Cellular Survival ◽  
The Family ◽  
Group A ◽  
Relationship Of ◽  
Hemoglobin Metabolism

Abstract 1. An hereditary hemolytic anemia is described which is normocytic and normochromic, transmitted as a mendelian dominant and not cured by splenectomy. In the family studied, the anemia was associated but not genetically linked with brachyphalangia. Acute idiopathic porphyria may also have been associated. All anemic members were of blood group A. 2. Neither the degree of anemia nor the rate of hemolysis was favorably influenced by splenectomy. 3. Various studies of erythrocyte fragility and hemoglobin metabolism are presented. 4. Although the red cells in this anemia were more resistant to fragility tests in vitro than the red cells of hereditary spherocytosis, they were more rapidly destroyed in vivo. 5. The disease is differentiated from other hereditary and hemolytic anemias. Of the hereditary anemias, this disease seems most closely to resemble hereditary spherocytosis. Yet the differences of cellular survival in vivo and in vitro and the failure of splenectomy in hereditary nonspherocytic hemolytic anemia suggest a difference in the hemolytic mechanism. 6. The demonstration of porphobilinogen in this patient suggests a possible relationship of this hereditary hemolytic anemia to hereditary porphyria.

scholarly journals Fragmentation and myelin formation in hereditary xerocytosis and other hemolytic anemias

Blood ◽  
1978 ◽  
Vol 52 (4) ◽  
pp. 750-761
Author(s):  
LM Snyder ◽  
HU Lutz ◽  
N Sauberman ◽  
J Jacobs ◽  
NL Fortier
Keyword(s):  
Hereditary Spherocytosis ◽  
Protein Composition ◽  
Atp Depletion ◽  
Red Cells ◽  
Integral Membrane Proteins ◽  
Propranolol Hydrochloride ◽  
Membrane Defects ◽  
Degree Of Fragmentation

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.

scholarly journals Physical Properties of Red Cells as Related to Effects in Vivo. III. Effect of Thermal Treatment on Survival of Red Cells in the Dog. Role of the Spleen

Blood ◽  
1968 ◽  
Vol 32 (6) ◽  
pp. 872-883 ◽  
Author(s):  
DENNIS J. CARLSON ◽  
THOMAS HALE HAM
Keyword(s):  
Sickle Cell Disease ◽  
Thermal Treatment ◽  
Peripheral Blood ◽  
Hereditary Spherocytosis ◽  
Osmotic Fragility ◽  
Red Cells ◽  
Cell Disease ◽  
Homozygous Sickle Cell Disease

Abstract Dog red cells heated at 49 C. for 15 minutes with no change in osmotic fragility, showed a decreased rate of survival in vivo, and increased sequestration by the spleen, and an increase in the osmotic fragility when recovered from the spleen. The peripheral blood showed normal osmotic fragility at all times. These changes were comparable to those seen in the spleen in homozygous sickle-cell disease and hereditary spherocytosis. There was no hemoglobinemia. Splenectomy decreased the rate of destruction of such heated red cells in vivo. In these studies the rigidity of the red cells with increased viscosity, but with normal osmotic fragility, may have resulted in the removal of the heated red cells by the spleen and subsequent conditioning of these cells in the spleen. In contrast to the 15 minutes heating, dog cells heated at 49 C. for 60 minutes had increased osmotic fragility and were rapidly removed from the circulation. There was sequestration by the spleen or by the liver with hemoglobinemia. The red cells with the greatest increase in osmotic fragility were not preferentially removed in vivo.

scholarly journals Fragmentation and myelin formation in hereditary xerocytosis and other hemolytic anemias

Blood ◽  
1978 ◽  
Vol 52 (4) ◽  
pp. 750-761 ◽  
Author(s):  
LM Snyder ◽  
HU Lutz ◽  
N Sauberman ◽  
J Jacobs ◽  
NL Fortier
Keyword(s):  
Hereditary Spherocytosis ◽  
Protein Composition ◽  
Atp Depletion ◽  
Red Cells ◽  
Integral Membrane Proteins ◽  
Propranolol Hydrochloride ◽  
Membrane Defects ◽  
Degree Of Fragmentation

Abstract 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.

scholarly journals Hemolytic Disease in Newborn Dogs

Blood ◽  
1951 ◽  
Vol 6 (4) ◽  
pp. 291-313 ◽  
Author(s):  
LAWRENCE E. YOUNG ◽  
RICHARD M. CHRISTIAN ◽  
DONALD M. ERVIN ◽  
R. WENDELL DAVIS ◽  
WILLIAM A. O’BRIEN ◽  
...  
Keyword(s):  
Osmotic Fragility ◽  
Red Cells ◽  
Rabbit Serum ◽  
Hemolytic Disease ◽  
Human Infants ◽  
Basal Nuclei ◽  
Intravenous Injections ◽  
Erythroid Hyperplasia ◽  
Dog Liver

Abstract 1. Bitches with erythrocytes lacking the canine A factor were immunized with intravenous injections of A-positive dog cells and mated with A-positive sires. ll A-positive pups born to such dams developed hemolytic disease provided they suckled the immunized dam during the first day of life. There was no evidence of transplacental isoimmunization or of transfer of antibody across the placenta from mother to pup. Anti-A could not be demonstrated in the blood of pups at birth and was not acquired by pups that first suckled an immunized bitch after the first day of life. A-positive pups born to a non-immunized bitch developed hemolytic disease after suckling an immunized foster dam on the day of birth. 2. Of 24 affected A-positive pups, 3 were sacrificed, 9 died within three days of birth and 12 recovered, 2 with the aid of transfusions of A-negative blood. Autopsies revealed varying degrees of hepatomegaly, splenomegaly, erythroid hyperplasia of the bone marrow, extra-medullary erythropoiesis, and questionable evidence of specific injury of the nerve cells of the basal nuclei of the brain. 3. The degree of anemia in the A-positive pups varied widely, the minimum hematocrit for the group being 10 per cent at forty-eight hours after birth. Erythroblastosis, reticulocytosis and spherocytosis were noted in most of the severely affected pups and osmotic fragility of the red cells was substantially increased in all of the A-positive pups exposed to anti-A. 4. The concentration of bilirubin in the serum of most of the affected pups was only slightly increased. The relatively small increases in serum bilirubin, compared with those in hemolytic disease of human infants, are presumably attributable to the unusual capacity of the dog liver for excreting bilirubin. 5. The red cells of all affected A-positive pups gave antiglobulin (Coombs) reactions and in surviving pups the cells were agglutinable in antiglobulin rabbit serum for as long as twenty-two days in A pups and sixty-five days in A’ pups. The degree of reactivity with antiglobulin serum was not correlated with the severity of the hemolytic process. Erythrocytes of the very mildly affected A’ pups were strongly agglutinated by antiglobulin serum but showed no definite sphering and only slight increase in osmotic fragility. 6. Serum of the 20 A-negative litter mates examined in this study contained anti-A for periods as long as thirty-two days while the red cells of these pups remained normal. A few of the A-positive pups also had anti-A in the serum. The in vitro behavior of anti-A in the pups’ sera was identical with that of anti-A in the maternal sera. 7. Twelve C-positive pups in 2 litters born to bitches immunized against the canine C factor showed no evidence of hemolytic disease. 8. Hemolytic disease of newborn dogs is compared with that of human infants and the need for further investigation of certain aspects of the disorder in both species is stressed.

scholarly journals Atypical Autohemolysis in Hereditary Spherocytosis as a Reflection of Two Cell Populations: Relationship of Cell Lipids to Conditioning by the Spleen

Blood ◽  
1968 ◽  
Vol 32 (4) ◽  
pp. 569-585 ◽  
Author(s):  
G. R. LANGLEY ◽  
C. H. FELDERHOF
Keyword(s):  
Hereditary Spherocytosis ◽  
Large Population ◽  
Osmotic Fragility ◽  
Cell Populations ◽  
Osmotic Resistance ◽  
Considerable Decrease ◽  
Cell Lipid ◽  
Relationship Of

Abstract Two patients with hereditary spherocytosis had increased autohemolysis not corrected by supplementary glucose, while an affected son of the second patient had a typical autohemolytic response with glucose. Almost one half of the circulating red cells in both patients had markedly increased osmotic fragility. When these were eliminated in vitro, by selective osmotic hemolysis, or in vivo by splenectomy, the autohemolysis response was corrected by glucose. The loss of this large population of extremely fragile cells after splenectomy suggested they were the product of conditioning by the spleen. In addition to having decreased osmotic resistance, an autohemolysis not corrected by glucose, and a short in vivo survival, these conditioned cells were smaller and had a considerable decrease in cell lipid. After splenectomy, with the loss of the most fragile cell population, cell size and lipid content increased. However in spite of no decrease in cell lipid after splenectomy, osmotic fragility remained abnormal. Increased osmotic fragility may be one of the earliest manifestations of a membrane abnormality in hereditary spherocytosis.

LACK OF EFFECT OF CORTICOSTEROIDS ON THE IN VIVO DISAPPEARANCE OF SULFHYDRYL-INHIBITED AND ANTIBODY-COATED ERYTHROCYTES

1964 ◽  
Vol 47 (3_Suppl) ◽  
pp. S28-S36
Author(s):  
Kailash N. Agarwal
Keyword(s):  
Red Cells ◽  
List Type ◽  
Red Cell

ABSTRACT Red cells were incubated in vitro with sulfhydryl inhibitors and Rhantibody with and without prior incubation with prednisolone-hemisuccinate. These erythrocytes were labelled with Cr51 and P32 and their disappearance in vivo after autotransfusion was measured. Prior incubation with prednisolone-hemisuccinate had no effect on the rate of red cell disappearance. The disappearance of the cells was shown to take place without appreciable intravascular destruction.

scholarly journals In Vitro and In Vivo Biosafety Analysis of Resorbable Polyglycolic Acid-Polylactic Acid Block Copolymer Composites for Spinal Fixation

Polymers ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 29
Author(s):  
Seung Kyun Yoon ◽  
Jin Ho Yang ◽  
Hyun Tae Lim ◽  
Young-Wook Chang ◽  
Muhammad Ayyoob ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Animal Experiments ◽  
Polymeric Materials ◽  
Polyglycolic Acid ◽  
Poly Lactic Acid ◽  
Skin Sensitization ◽  
Spinal Fixation ◽  
In Vivo Degradation

Herein, spinal fixation implants were constructed using degradable polymeric materials such as PGA–PLA block copolymers (poly(glycolic acid-b-lactic acid)). These materials were reinforced by blending with HA-g-PLA (hydroxyapatite-graft-poly lactic acid) and PGA fiber before being tested to confirm its biocompatibility via in vitro (MTT assay) and in vivo animal experiments (i.e., skin sensitization, intradermal intracutaneous reaction, and in vivo degradation tests). Every specimen exhibited suitable biocompatibility and biodegradability for use as resorbable spinal fixation materials.

scholarly journals The impact of indole-3-lactic acid on immature intestinal innate immunity and development: a transcriptomic analysis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Wuyang Huang ◽  
Ky Young Cho ◽  
Di Meng ◽  
W. Allan Walker
Keyword(s):  
Lactic Acid ◽  
Mechanistic Model ◽  
Transcriptomic Analysis ◽  
Dependent Manner ◽  
Protective Effects ◽  
Very Preterm Infants ◽  
Anti Inflammatory ◽  
The Impact

AbstractAn excessive intestinal inflammatory response may have a role in the pathogenesis of necrotizing enterocolitis (NEC) in very preterm infants. Indole-3-lactic acid (ILA) of breastmilk tryptophan was identified as the anti-inflammatory metabolite involved in probiotic conditioned media from Bifidobacteria longum subsp infantis. This study aimed to explore the molecular endocytic pathways involved in the protective ILA effect against inflammation. H4 cells, Caco-2 cells, C57BL/6 pup and adult mice were used to compare the anti-inflammatory mechanisms between immature and mature enterocytes in vitro and in vivo. The results show that ILA has pleiotropic protective effects on immature enterocytes including anti-inflammatory, anti-viral, and developmental regulatory potentials in a region-dependent and an age-dependent manner. Quantitative transcriptomic analysis revealed a new mechanistic model in which STAT1 pathways play an important role in IL-1β-induced inflammation and ILA has a regulatory effect on STAT1 pathways. These studies were validated by real-time RT-qPCR and STAT1 inhibitor experiments. Different protective reactions of ILA between immature and mature enterocytes indicated that ILA’s effects are developmentally regulated. These findings may be helpful in preventing NEC for premature infants.

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