Shortened Erythrocyte Lifespan Research Articles

The effect of endoscopic retrograde cholangiopancreatography on the serum IL-18 and erythrocytes antioxidative capacity in biliary obstructive jaundice

Introduction: Obstructive jaundice is an important clinical problem. It may cause transient hemolysis and shortened erythrocyte life span as well as cytokine induction. An increase in lipid peroxidation has been noted as evidence of oxidative damage in red cells due to cholestasis. The influence of endoscopic retrograde cholangiopancreatography (ERCP), mechanical lithotrepsy and stone extraction on the antioxidative capacity of the erythrocyte and immune response is still unclear. Methods: Superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) content of red blood cells (RBC), and serum interleukin (IL-18) were measured in 20 patients with calcular obstructive jaundice before and 4 weeks after ERCP intervention and compared with 10 matched healthy volunteers. Results: A significant decrease ( p<0.05) in SOD and CAT activities and glutathione concentration but a significant increase ( p<0.05) in serum IL-18 were observed in cholestatic patients compared with the healthy control and were significantly correlated with variable of hepatic dysfunction (alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, alkaline phosphatase (ALP) and gamma glutamyl transferase (GGT). After ERCP, serum IL-18 and antioxidant capacity of red blood cells were significantly improved and returned to normal concentration ( p<0.05). Conclusions: In biliary obstruction, serum IL-18 is increased and antioxidative capacity is decreased, and have a direct correlation with biochemical markers of liver injury. After ERCP intervention, the altered antioxidative capacity as well as serum IL-18 was completely restored to normal.

Can Glycohemoglobin Be Used to Assess Glycemic Control in Patients with Chronic Renal Failure?

Many factors can affect interpretation of glycohemoglobin (GHB/HbA1c) measurements in patients with chronic renal failure (CRF). Several reports have suggested that erythrocyte survival is substantially lowered in most patients with CRF; this would be expected to lower GHB results (1)(2)(3)(4)(5)(6). Several reports have also suggested that GHB methods, especially those based on charge separation (e.g., ion-exchange HPLC), may have interference by carbamylated hemoglobin that would be expected to falsely increase GHB results (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). Many of these reports evaluated older assay methods; newer ion-exchange methods may show improved separation of the HbA1c fraction from other hemoglobin adducts (15)(17). Because renal failure is common in patients with diabetes and GHB is widely used as an index of mean blood glucose in these patients, we examined GHB results in patients with CRF by several different GHB assay methods. We also investigated the impact of shortened erythrocyte lifespan by comparing the GHB results obtained for nondiabetic patients with and without CRF. Fifty-five patients with CRF (blood urea nitrogen >400 mg/L) were recruited for this study. Twenty-nine were not receiving dialysis and …

Involvement of phosphatidylserine exposure in the recognition and phagocytosis of uremic erythrocytes

Cell surface-exposed phosphatidylserine (PS) represents a signal for macrophage recognition and cell phagocytosis. This study examines PS exposure and susceptibility to erythrocyte phagocytosis in patients with chronic uremia in an attempt to assess the possible pathogenic mechanism behind cell removal in a condition associated with shortened erythrocyte life. Both PS-expressing erythrocytes and erythrophagocytosis (human monocyte-derived macrophages ingesting one or more erythrocytes) were significantly increased in uremic patients compared with healthy controls. Phagocytosed uremic erythrocytes appeared intact, suggesting they were identified before lysis through some surface change recognized by the macrophages. The degree of phagocytosis was markedly greater for PS-positive than PS-negative fluorescence-activated cell sorter (FACS)-sorted uremic erythrocytes. A significant correlation (r = 0.655) was found between the percentage of PS-expressing red blood cells (RBCs) and the percentage of phagocytosing macrophages in uremic patients. Reconstitution experiments showed the ability of uremic plasma to promote both PS exposure and erythrophagocytosis, the latter without direct interaction with the macrophage population. Phagocytosis of uremic erythrocytes was strongly inhibited when the macrophages were preincubated with glycerophosphorylserine (GPS), a structural derivative of PS, but this was not the case with the equivalent derivative of phosphatidylethanolamine, glycerophosphorylethanolamine. This inhibition appeared to be specific because GPS failed to inhibit the phagocytosis of opsonized uremic erythrocytes that occurs through an Fc receptor-mediated pathway. These findings suggest that a PS-recognition mechanism may promote the susceptibility of uremic RBCs to phagocytosis and thus be involved in the shortened erythrocyte life span of uremia.

Glutathione Regeneration in Mammalian Erythrocytes

Glutathione (GSH) regeneration is a process in which cells reduce oxidised glutathione (GSSG) to GSH after exposure of cells to oxidants in the presence of suitable energy source such as glucose. This reaction consists of (1) membrane glucose transport, (2) phosphorylation of glucose by hexokinase (HK) utilising adenosine 5′-triphosphate (ATP), (3) reduction of nicotinamide–adenine dinucleotide phosphate (NADP) to its reduced form (NADPH) by glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phosphogluconate dehydrogenase (6-PGD) and (4) reduction of GSSG to GSH by glutathione reductase (GR) using NADPH. The rate of GSH regeneration is thus dependent on the enzymatic activity and concentration of substrate. G-6-PD deficiency and enzyme inhibitory chemicals reduce GSH regeneration and concentration of substrate greatly influences the rate of GSH regeneration. Not only glucose but also mannose, fructose and galactose may also be used as energy source. Interspecies and intraspecies differences occur in the rate of GSH regeneration. These differences cannot be explained by variations in enzymatic activities. Although physiological relevance of GSH regeneration is still unclear, it is known that erythrocytes from G-6-PD-deficient patients have lowered erythrocyte deformability and shortened erythrocyte life-span. In-vitro experiments show lowered GSH regeneration enforces loss of deformability. These findings suggest that GSH regeneration plays an important role in erythrocyte biology.

Hereditary elliptocytosis with protein band 4.1 deficiency in the dog

A dog with persistent elliptocytosis was studied. The dog had membrane protein band 4.1 deficiency, microcytosis, shortened erythrocyte lifespan, increased osmotic sensitivity, and a mild glutathione deficiency. Erythrocyte deformability and membrane stability were adversely effected. The dog's parents had decreased band 4.1, decreased stability, and some elliptocytosis. This disorder in dogs closely resembles human patients with band 4.1 deficiency and should provide a valuable animal model to study the erythrocyte membrane cytoskeleton.

Hereditary Elliptocytosis With Protein Band 4.1 Deficiency in the Dog

A dog with persistent elliptocytosis was studied. The dog had membrane protein band 4.1 deficiency, microcytosis, shortened erythrocyte lifespan, increased osmotic sensitivity, and a mild glutathione deficiency. Erythrocyte deformability and membrane stability were adversely effected. The dog's parents had decreased band 4.1, decreased stability, and some elliptocytosis. This disorder in dogs closely resembles human patients with band 4.1 deficiency and should provide a valuable animal model to study the erythrocyte membrane cytoskeleton.

Further evidence for oxidative damage to hemoglobin and red cell membrane in leukemia

The presence of significant methemoglobinemia in a large number (79%) of cases of leukemia prompted us to look into the evidence for oxidative injury to the other red cell constituents, e.g. the cell membrane. Forty-five per cent of cases showed increased malonyldialdehyde (MDA) levels indicating in vivo peroxidation of membrane lipids in leukemia patients. Red cell superoxide dismutase (SOD) activity was reduced in 25% of cases while plasma α-tocopherol levels were markedly low in 68% of patients. The presence of an inverse correlation between MDA levels and SOD and tocopherol levels in a large number of cases suggested that the red cells in leukemia lack the ability to counter increased oxidative stress. Low hemoglobin content of the red cells in these patients seemed to contribute to the oxidative injury to the membrane by rendering the latter more accessible to the oxidants. While methemoglobinemia may enhance tissue hypoxia, membrane lipid peroxidation could explain the shortened erythrocyte life-span and anemia in leukemia.

Oxidative haemolysis in protein malnutrition

A study of the haemolytic anaemia observed in protein-energy malnutrition (PEM) in Kivu disclosed the following results. The in vitro resistance to oxidative aggressions of PEM patients' erythrocytes was decreased: when incubated with acetylphenylhydrazine, a higher percentage of the cells showed Heinz bodies, as compared with erythrocytes of local controls. Normal or increased activities were found for certain erythrocyte enzymes involved in the detoxification of activated oxygen: glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase. The level of reduced glutathione was not decreased. Reduced activities were observed for two enzymes containing trace elements: glutathione peroxidase and Superoxide dismutase. It is suggested that the shortened erythrocyte lifespan observed in PEM patients corresponds to an oxidative process which results from the decrease of both enzyme activities. The hypothesis that depletion of trace elements could be responsible for the decreased activity of those enzymes is discussed.

The erythrocyte membrane disturbances in protein-energy malnutrition: nature and mechanisms.

In protein-energy malnutrition (PEM), as observed in Kivu, the RBC have an increased ratio of surface area to volume which is demonstrated by the presence of target cells on light microscopy and cup cells with scanning electron microscopy. The osmotic fragility is decreased. These abnormalities can be attributed to the accumulation of cholesterol and phosphatidylcholine (PC) in the RBC membrane. The molar ratio of cholesterol to phospholipids is moderately increased. Several findings suggest that the cholesterol and PC build-up results from disturbed exchanges in these lipids between the RBC and the plasma lipoproteins. Firstly, the osmotic fragility of a patient's RBC gradually becomes normal when the cells are transfused into a healthy recipient. Secondly, the cholesterol flux between the RBC and the plasma LDL seems to be low. Thirdly, the increase in RBC PC cannot be explained by a diminished fatty acids transport between the deep RBC PC pool and the RBC phosphatidylethanolamine (PE) pool. Finally complex disturbances of the plasma lipoproteins are obvious. It is improbable that the cholesterol and PC build-up accounts for the premature RBC destruction which has been described in Kivu PEM. However, the observation of an increased fatty acid turnover in RBC PC and PE, as well as other data previously obtained in Kivu PEM, lead to the conclusion that membrane peroxidation may be a major cause of the shortened erythrocyte life-span in this syndrome.

Red cell mass: Its precursors and its perturbations.

There are only two fundamental causes of anemia: diminished erythrocyte production and shortened erythrocyte life span. Genetic, environmental, pathologic, and iatrogenic factors that may reduce red cell mass are put in clinical perspective.

The effects of polymeric plutonium on erythrocyte survival in mice. II. Changes in the RES function and blood volume.

The function of reticuloendothelial system (RES) to sequestrate 51Cr labelled heat damaged erythrocytes was investigated in male CF#1 mice after intravenous injection of polymeric plutonium at the dose levels of 15 μCi/kg, 10μCi/kg and 5μCi/kg. Effect of splenectomy on erythrocyte life span was also examined. Total blood volume, red cell volume and plasma volume were also determined so as to study the possible cause of anemia induced by polymeric plutonium. It was observed that polymeric plutonium depressed RES function of the spleen to sequestrate 51Cr labelled heat damaged erythrocytes, but disappearance of 51Cr labelled heatdamaged erythrocytes from the circulation of plutonium-exposed mice was within normal range. Erythrocyte life span was unchanged after splenectomy. Normal disappearance rate of 51Cr labelled heat-damaged erythrocytes from the circulation with depressed RES function of the spleen by polymeric plutonium was considered to be the result of increased function of the other parts of RES. In higher dose groups, red cell volume was decreased but no change in plasma volume was observed in any group studied. From these results, the possible cause of the shortened erythrocyte life span and anemia induced by extracorpuscular effect of polymeric plutonium are discussed. It is concluded at this time that decreased RES function of the spleen may not play an important role in shortened erythrocyte life span.

The biosynthesis of human hemoglobin A1c. Slow glycosylation of hemoglobin in vivo.

Hemoglobin A1c, the most abundant minor hemoglobin component in human erythrocytes, is formed by the condensation of glucose with the N-terminal amino groups of the beta-chains of Hb A. The biosynthesis of this glycosylated hemoglobin was studied in vitro by incubating suspensions of reticulocytes and bone marrow cells with [3H]leucine or 59Fe-bound transferrin. In all experiments, the specific activity of Hb A1c was significantly lower than that of Hb A, suggesting that the formation of Hb A1c is a posttranslational modification. The formation of Hb A1c in vivo was determined in two individuals who were given an infusion of 59Fe-labeled transferrin. As expected, the specific activity of Hb A rose promptly to a maximum during the 1st week and remained nearly constant thereafter. In contrast, the specific activity of Hb A1c and also of Hbs A1a and A1b rose slowly, reaching that of Hb A by about day 60. These results indicate that Hb A1c is slowly formed during the 120-day life-span of the erythrocyte, probably by a nonenzymatic process. Patients with shortened erythrocyte life-span due to hemolysis had markedly decreased levels of Hb A1c.

Erythrocyte Life Span and Hemoglobin Levels in Mouse Trichuriasis

The erythrocyte life span, hemoglobin levels, and blood loss in the urine and feces of mice were determined during infection with Trichuris muris. Hemoglobin levels indicated only a minor transitory erythropoietic response at the height of patency. The mean apparent half-times of circulating erythrocytes in 2 groups of normal mice, and in those with estimated mean worm burdens of 82 and 92 were, respectively, 18.5, 19.3, 18.8, and 15.8 days. Through day 21 normal mice excreted 28.9% of the total administered 'Cr, 26.5% in urine, 2.4% in feces. Combined excreta of infected mice gave similar values. 'Cr radioactivity statistically greater than that of the background count could not be detected in a suspension of 562 worms obtained 48 hr after the hosts had received the labeled erythrocytes. It is concluded that hemorrhage or marked blood loss was not a factor in the observed slightly shortened erythrocyte life span in mice with a mean worm burden of 92 ? 23 worms. Trichuris trichiura infection in man is frequently accompanied by bloody diarrhea as is T. vulpis in dogs (Miller, 1939); T. ovis in sheep (Powers, 1961); and T. suis in swine (Powers et al., 1960). In a study of human trichuriasis Layrisse et al. (1967) found that patients lose about 0.8 ml of blood per million eggs per day. Other reports have suggested that T. trichiura may, or may not, be responsible for the concomitant anemia frequently observed in this parasitosis: Guiart (1908), Christoffersen (1914), Swartzwelder (1939), Jaffe (1945), Hartz (1953), Guillon (1953), Gajardo and Atias (1956), Briscoe (1957), Winship and Hennessy (1959), Boon and Hoh (1961), Silverston (1962), Ramsey (1962), Martinez-Marafi6n et al. (1968). Burrows and Lillis (1964) obtained a positive benzidine reaction with T. vulpis body fluid released onto paper and suggested that this reaction along with their finding of unmistakable evidence of having blood cells in the esophageal lumen indicated that whipworms are . . . avid blood suckers. Hemoglobins, present in several nematodes, have been shown to differ spectroscopically from those of the host (Rogers, 1962; Smith and Lee, 1963). Received for publication 23 July 1970. * This study was supported in part by the U. S. Public Health Service Training Grant 2E-55 (C2 and C3). t Present address: Department of Microbiology, New York Medical College, 5th Avenue and 106th Street, New York, N. Y. 10029. Efremov and Shikhobalova (1939) stated that T. muris does not suck blood; it secretes a proteolytic ferment which digests the surrounding tissues. The present studies were designed to investiga e the erythrocyte life span, blood loss, and hemoglobin levels of mice infected with T. muris. MATERIALS AND METHODS 1) Culture and infection. The strain of T. muris, preparation of egg cultures, and method of inoculation were as previously described (Pike, 1969). 2) Experimental animals. The DBA/2 weanling mice were raised by the author through brothersister matings of Jackson Laboratory stock DBA/2J mice. Initially 112 male and 48 female mice were used. They were randomly divided into 4 groups: Group 1. controls, 20 males Group2. each given 150 T. muris eggs, 30 males, 14 females Group 3. each given 300 T. muris eggs, 32 males, 14 females Group 4. blood donors, 30 males, 20 females. Inoculated mice which did not develop patent infections were subsequently discarded. All patent female mice of groups 2 and 3 and males which at the end of the 2nd week of patency had 24-hr fecal egg counts of 30,000 or less were used as a source of worms to be subjected to Cr counting. The experimental groups were ultimately composed of: Group 1. controls, 20 males Group 2. 150 eggs, 17 males Group 3. 300 eggs, 17 males. 3) Labeling of red blood cells. Na25CrO4 (E. R. Squibb and Sons) with a specific activity of 92.3 mc/mg of metallic chromium was employed. The solution was added to 25 ml of pooled heparinized blood from group 4 mice to yield 5 ,uc/0.25 ml, the

ANEMIA IN UREMIA

Abstract. One hundred and thirty‐eight patients with renal insufficiency of varying degree have been studied with respect to anemia and signs of hemolysis. A control material is included in the study. 1) The patient group displayed a normochromic anemia with only a slightly subnormal mean corpuscular hemoglobin concentration. 2) A moderately elevated reticulocyte count together with a tendency to elevated endogenous carbon monoxide production indicated a certain degree of hemolysis in the uremic patient group compared with the controls. Patients with chronic pyelonephritis and active urinary infection displayed an elevated reticulocyte count (p &lt; 0.001) and lower haptoglobin values (0.05 &gt; p &gt; 0.01) than non‐infected uremic patients. Thus we consider these infections to be a hemolysis‐promoting factor in patients with renal insufficiency. 3) In men there was a positive correlation between the concentrations of serum creatinine versus serum iron concentration and the saturation percentage of the total iron binding capacity. In the light of cited iron kinetic data, we consider this correlation an indicator of depressed erythropoiesis. 4) A selected group of severely ill uremic patients were studied with respect to their erythrocyte life span by the bilirubin turnover method. As a group, they displayed a shortened erythrocyte life span in comparison with an earlier published control group.

Anemia in macroglobulinemia

Erythrokinetic and red cell survival studies were performed in ten patients with macroglobulinemia. Inadequate red cell synthesis, decreased erythrocyte survival and iron deficiency, alone or in combination, were responsible for anemia in eight of these patients. In six patients there was a relative failure of red cell production with inadequate compensation for excessive red cell destruction. In two patients there was an absolute failure of erythropoiesis with functional aplasia. No simple correlation between rate of erythropoiesis and serum gamma macroglobulin concentration was evident. Shortened erythrocyte life span was found in eight of the ten patients and was most prominent in those with active disease. Serial studies before and after plasmapheresis in two patients showed that reduction of serum viscosity and macroglobulin level was associated with increased red cell survival. Causes of shortened erythrocyte survival included bleeding disorders, Coombs' positive hemolytic anemia and erythrocyte sequestration by the spleen. Two patients had advanced iron deficiency and four had evidence of early iron deficiency. Depletion of body iron stores usually could be related to external blood loss. The typical features of iron depletion were present when iron deficiency was advanced. Therapy of anemia in these patients consisted of plasmapheresis with or without chemotherapy for disease manifestations associated with high serum viscosity, corticosteroids when immune hemolysis was demonstrable, and iron for iron deficiency.

Studies of the production, volume and destruction of erythrocytes in patients with sarcoidosis.

Previous studies in patients with diseases characterized by chronic inflammation have shown abnormalities in both iron metabolism and erythrocyte life span. Patients with acute rheumatic fever may have a shortened erythrocyte life span (I). Patients with chronic infection or rheumatoid arthritis have been shown to have anemia (2-9), hypoferremia (8-11), and a normal (5-7) or shortened (3, 4, 8, 12, 13) erythrocyte life span. It is the purpose of this report to present an investigation of the iron metabolism and apparent erythrocyte life span in patients with sarcoidosis.

Acetylcholinesterase and the life-span of the erythrocyte.

THE function of the relatively large amount of the enzyme acetylcholinesterase which is present in erythrocytes is unknown. The enzyme was thought to be essential for the maintenance of normal permeability of the erythrocyte1, but experiments on cells deficient in acetylcholinesterase2 have failed to demonstrate this function. Interest in red-cell acetylcholinesterase has been stimulated by the recent finding that in some patients with paroxysmal nocturnal haemoglobinuria (P.N.H.) the enzyme is deficient3. The enzyme activity in patients with P.N.H. varies from 6–134 per cent of average normal activity4, and there is a correlation between the degree of enzyme deficiency and some other features of P.N.H.4. It is thus possible that the shortened erythrocyte life-span in P.N.H. may be related to the enzyme deficiency, and that normal amounts of acetylcholinesterase are necessary for normal survival of erythrocytos.

HEREDITARY ELLIPTOCYTOSIS ASSOCIATED WITH INCREASED HEMOLYSIS

Three cases of hereditary elliptocytosis have been presented. The criteria for diagnosis have been (1) the presence of a high percentage of elongated cells in stained smears and wet preparations, and (2) the finding of elongated cells in other members of the family including one parent in each case. All 3 children had a marked anemia when first seen. Case 1 had an outspoken hemolytic anemia at 2 months of age. His hemoglobin is now practically normal, but he still has a slight but persistent reticulocyte increase. Cases 2 and 3 were first thought to be due to iron deficiency anemia. While iron deficiency may have been responsible for the degree of anemia that brought the cases under observation, a slight but persistent elevation in both reticulocyte count and stool urobilin concentration indicated the presence of increased hemolysis. The findings of a shortened erythrocyte life span in Case 3 supports this conclusion. A review of the literature makes it clear that hereditary elliptocytosis is a potentially hemolytic disease. However, in the great majority of the cases the elliptocytosis appears as a harmless anomaly without pathological significance. In a certain number, to which considerable attention is now being paid, there may be a hemolytic anemia sometimes of a severity sufficient to require splenectomy. Between these 2 extremes there lies an indeterminate group in which there is evidence of hemolysis if one looks for it, in which there may or may not be anemia, and in which it is not always easy to know if the patient's symptoms are part of the disease. It is to this group that our patients belong.