RESUMEN
BACKGROUND: Irradiation has been shown to adversely affect both in vivo 24-hour recovery (recovery [%]) and in vitro properties of stored red cells (RBCs). There is uncertainty as to how these changes are related to the day of irradiation and the length of storage after irradiation. STUDY DESIGN AND METHODS: Four protocols used day of irradiation and storage time after irradiation as the independent variables. At the conclusion of the storage period, viability was measured with radiolabeled RBCs as the recovery and the long-term survival time for RBCs that were circulating beyond 24 hours. In addition, in vitro values including RBC ATP, hemolysis level, and supernatant potassium were measured. Each subject donated 2 units of whole blood (CPD) and received autologous irradiated and untreated control RBCs (AS-1) on two separate occasions. RESULTS: Reduced recovery in irradiated units was noted when compared to that in control units, and the reduction was most apparent with long periods of storage after irradiation, irrespective of the day of irradiation. With irradiation on Day 1 of storage and a total storage period of 28 days, mean +/- SD recovery (single label) was 84.2 +/- 5.1 percent for control RBCs and 78.6 +/- 5.9 percent for irradiated RBCs (n = 16; p<0.01). With irradiation on Day 14 and storage through Day 42, the recoveries were 76.3 +/- 7.0 percent for control RBCs and 69.5 +/- 8.6 percent for irradiated RBCs (n = 16; p<0.01). Less reduction in recovery was observed with shortening of the postirradiation storage time. When the total storage period was reduced to 28 days after Day 14 irradiation, the recoveries were not significantly different. With an additional 2-day storage period after irradiation on Day 26, the recoveries were also comparable. Long-term survival times for control and irradiated RBCs were not significantly different in any of the four protocols. RBC ATP levels and hemolysis were minimally, but significantly influenced by irradiation. Supernatant potassium levels, however, were substantially increased after irradiation in each of the four protocols. CONCLUSION: Irradiation has only a small effect on the properties of RBCs treated and stored according to the utilized protocols. Longer storage times after irradiation resulted in progressively reduced recovery while long-term survival remained unaffected.
Asunto(s)
Conservación de la Sangre , Envejecimiento Eritrocítico/efectos de la radiación , Rayos gamma , Transfusión Sanguínea , Estudios Cruzados , Hemólisis/fisiología , Humanos , Técnicas In Vitro , Potasio/sangreRESUMEN
BACKGROUND: Cellular blood components are irradiated to prevent graft-versus-host disease in transfusion recipients at risk for this syndrome. Because gamma radiation can result in the production of reactive oxygen species, the role of reactive oxygen species was investigated in radiation-induced red cell damage. STUDY DESIGN AND METHODS: Whole blood from normal donors was exposed to various doses of t-butyl hydroperoxide (0-1 mM) and/or to gamma-radiation (0-50 Gy). Oxidative damage was assessed by the extent of lipid peroxidation (measured by thiobarbituric acid-reactive substances [TBARS]) and hemoglobin oxidation. Fresh blood was divided into three parts-one initially irradiated and stored, another stored with portions irradiated weekly, and a third stored without irradiation. TBARS and hemoglobin oxidation were measured weekly. RESULTS: As expected, t-butyl hydroperoxide induced TBARS formation and hemoglobin oxidation in a dose-dependent fashion. The gamma-radiation not only increased hemoglobin oxidation and TBARS formation, but also enhanced the t-butyl hydroperoxide effect on red cells. Red cell storage increased TBARS generation and hemoglobin oxidation in a time-dependent fashion. When radiation was administered either initially or after weekly storage, TBARS production and hemoglobin oxidation were increased over that measured in unirradiated paired controls. CONCLUSION: Gamma radiation at clinically used doses increases lipid peroxidation and hemoglobin oxidation in human red cells. The effect of gamma-radiation is accentuated by blood storage and induces damage independent of time of storage.
Asunto(s)
Envejecimiento Eritrocítico/efectos de la radiación , Eritrocitos/efectos de la radiación , Traumatismos por Radiación , Antioxidantes/farmacología , Hidroxitolueno Butilado/farmacología , Rayos gamma , Hemoglobinas/metabolismo , Humanos , Peroxidación de Lípido/fisiología , Oxidación-Reducción , Traumatismos por Radiación/sangre , Especies Reactivas de Oxígeno/fisiologíaRESUMEN
BACKGROUND: Gamma irradiation of blood components is used to prevent transfusion-associated graft-versus-host disease. The demand for irradiated blood components is increasing because of the increase in directed donation by family members. Irradiated units currently have a recommended maximum storage life of 28 days. Since in vivo recovery is related to red cell ATP levels, rejuvenation of stored irradiated units using a pyruvate-inosine phosphate-adenine additive was explored. STUDY DESIGN AND METHODS: Units of AS-1 red cells from 16 volunteer donors were divided into two equal volumes and one split unit from each was irradiated with 25 Gy. Ten units were irradiated on Day 5, 6, or 7 of 4 degrees C storage and 6 units were irradiated on Day 1 of 4 degrees C storage. All units were rejuvenated for 1 hour at 37 degrees C using a pyruvate-inosine-phosphate-adenine additive on Day 42 of 4 degrees C storage. Units were assayed for ATP, 2, 3 DPG and supernatant sodium, potassium, and glucose. RESULTS: ATP and 2, 3 DPG levels were restored equally well in irradiated and non-irradiated units. The previously reported irradiation-induced red cell potassium-sodium shift was demonstrated. Supernatant potassium and sodium levels did not reverse 1 hour after rejuvenation was completed. There was no significant difference in results between units irradiated on Day 1 or Day 5, 6, or 7. CONCLUSION: Red cell ATP and 2, 3 DPG levels were restored in irradiated AS-1 units stored at 4 degrees C for 42 days using a pyruvate-inosine-phosphate-adenine rejuvenation additive.
Asunto(s)
Envejecimiento Eritrocítico/efectos de la radiación , 2,3-Difosfoglicerato , Adenosina Trifosfato/sangre , Glucemia/análisis , Ácidos Difosfoglicéricos/sangre , Eritrocitos/química , Rayos gamma , Humanos , Potasio/sangre , Sodio/sangre , Factores de TiempoRESUMEN
The oxidative modification of low density lipoprotein (LDL) has been implicated as an early step in the formation of atheromatous lesions. In vitro studies suggest it to be accelerated, or even initiated, by transition metals such as iron or copper in combination with a reducing agent. Even if such metals have been demonstrated in atheroma gruels, their origin and precise localisation within human atheroma are presently unknown. In the initial part of this study we applied Pearl's method, energy dispersive X-ray microanalysis, and a modified Timm sulphide silver method (SSM) to demonstrate the occurrence of iron in early atherosclerotic lesions from a number of consecutive autopsy cases with evident, general atheromatosis. With the very sensitive SSM, but not with the other techniques, we found foam cells to contain heavy metals with a mainly lysosomal localization. On the basis of the hypothesis that such a lysosomal accumulation of iron might be due to erythrophagocytosis by migrating tissue-bound macrophages that later develop into foam cells, we designed an in vitro model system where human monocyte-derived macrophages were exposed to artificially aged, UV-exposed erythrocytes. The macrophages were then exposed to LDL in serum-and iron-free RPMI medium, occasionally in the presence of the potent iron-chelator desferrioxamine. The capacity of macrophages to oxidise LDL was much enhanced following erythrophagocytosis, and the process was shown to involve secretion of iron. Consequently, LDL oxidation was greatly inhibited by desferrioxamine. We conclude that iron may be exocytosed by macrophages that previously had their lysosomal apparatus enriched with iron, e.g. due to erythrophagocytosis. Oxidation of LDL may result in ensuing foam cell-formation secondary to scavenger-receptor mediated endocytosis by macrophages.
Asunto(s)
Arteriosclerosis/metabolismo , Eritrocitos/química , Células Espumosas/química , Hierro/análisis , Lipoproteínas LDL/metabolismo , Macrófagos/fisiología , Fagocitosis , Anciano , Anciano de 80 o más Años , Animales , Enfermedades de la Aorta/metabolismo , Enfermedades de la Aorta/patología , Arteriosclerosis/etiología , Línea Celular , Células Cultivadas , Enfermedad de la Arteria Coronaria/metabolismo , Enfermedad de la Arteria Coronaria/patología , Medio de Cultivo Libre de Suero , Envejecimiento Eritrocítico/efectos de la radiación , Eritrocitos/efectos de la radiación , Exocitosis , Femenino , Compuestos Ferrosos/farmacología , Humanos , Hierro/efectos adversos , Lisosomas/química , Macrófagos/efectos de los fármacos , Masculino , Ratones , Persona de Mediana Edad , Oxidación-Reducción , Especies Reactivas de OxígenoRESUMEN
Irradiation of blood components eliminates the risk of transfusion-associated graft-versus-host disease. Freezing directed or rare red cell units that are irradiated but not transfused would facilitate inventory management and would increase the transfusion options for the involved patients. However, no studies have been performed to evaluate whether prestorage irradiation damages subsequently frozen red cells. Ten normal volunteers donated a unit of whole blood on two separate occasions. One unit was irradiated with 15 Gy (1500 rad), stored at 4 degrees C for 6 days, and then frozen and stored at -75 degrees C for 56 days. The other unit (control) was similarly stored but was not irradiated. Aliquots of the units were tested on Day 0 and Day 6 and, after deglycerolization, on Day 62. Comparison of means and changes in means showed no significant differences in red cell ATP, 2,3 DPG, or supernatant hemoglobin and glucose in control and irradiated units. The difference in the change in supernatant potassium from Day 0 to Day 6 in control and irradiated units was significant (1.5 to 28.6 mmol/L vs. 1.5 to 48.5 mmol/L: p < 0.0001). Irradiation did not cause significant differences in postdeglycerolization red cell recovery (control, 84.5% vs. irradiated, 81.2%) or in 24-hour posttransfusion autologous red cell survival (control, 91.1% vs. irradiated, 90.9%). Red cells can be irradiated, stored at 4 degrees C for 6 days, and subsequently frozen with no increase in detectable damage as compared to controls that were not irradiated.
Asunto(s)
Conservación de la Sangre , Criopreservación , Envejecimiento Eritrocítico/efectos de la radiación , Eritrocitos , Radioisótopos de Cromo , Humanos , Marcaje Isotópico , Factores de TiempoRESUMEN
Transfusion-associated graft-versus-host disease (TA-GVHD) may occur whenever immunologically competent allogeneic lymphocytes are transfused to an immunocompromised recipient. Irradiation of blood components eliminates the risk of TA-GVHD but may damage the cellular elements in the transfused component, particularly if the cells are stored for prolonged periods in the irradiated state. To study the effect of irradiation on long-term storage of red cells, AS-1 red cells from eight normal subjects were prepared on two occasions. On one occasion, the units were stored as standard AS-1 red cells for 42 days at 4 degrees C; on the other, they were exposed to 3000 cGy radiation within 4 hours of collection and then were stored as AS-1 red cells for 42 days at 4 degrees C. The donations were at least 12 weeks apart. Irradiated units demonstrated significant elevations in poststorage plasma hemoglobin (Hb) (623 +/- 206 vs. 429 +/- 194 g/dL [6230 +/- 2060 vs. 4290 +/- 1940 g/L], p less than 0.02) and plasma potassium (78 +/- 4 vs. 43 +/- 9 mEq/L [78 +/- 4 vs. 43 +/- 9 mmol/L], p less than 0.01) and significant decreases in red cell ATP (1.9 +/- 0.2 vs. 2.1 +/- 0.3 microM/g Hb, p less than 0.04) and 24-hour posttransfusion red cell recovery (68.5 vs. 78.4%, p less than 0.02), as compared to nonirradiated units. It can be concluded that irradiation with 3000 cGy damages red cells and that long-term storage in the irradiated state may enhance this damage. Red cells should not be stored for 42 days after irradiation with 3000 cGy.
Asunto(s)
Conservación de la Sangre , Transfusión Sanguínea , Eritrocitos/efectos de la radiación , Envejecimiento Eritrocítico/efectos de la radiación , Rayos gamma , Humanos , Factores de TiempoRESUMEN
Dogs given total-body irradiation and marrow transplants from DLA-identical littermates exhibit prompt and sustained hematopoietic engraftment. However, animals given three preceding blood transfusions from the marrow donor before transplant become sensitized and reject the marrow graft. Rejection is due to exposure to polymorphic minor non-DLA histocompatibility antigens expressed on blood mononuclear cells. We sought to determine whether heat treatment would prevent blood from sensitizing recipients in this model since heating blood to 45 degrees C for 45 min abrogates the ability of blood mononuclear cells to stimulate in mixed lymphocyte culture. Three of 4 evaluable dogs given heat-treated blood before transplant rejected their marrow grafts. To prevent possible reexpression/reacquisition of mononuclear cell functional activity in vivo after transfusion, subsequent dogs were given heated blood that was additionally exposed to 2000 cGy gamma irradiation. Eight of 10 evaluable dogs given blood treated in this fashion engrafted. Unexpectedly, 9 out of 10 evaluable dogs transfused with blood treated only with gamma irradiation also engrafted. These results demonstrate that treatment of blood with gamma irradiation alone or in combination with heat prevents transfusion-induced sensitization to minor histocompatibility antigens. Results from this canine model suggest that blood products be gamma irradiated before transfusion in patients who are transplant candidates in order to prevent sensitization to minor histocompatibility antigens and reduce the risk of marrow graft rejection.
Asunto(s)
Transfusión Sanguínea , Trasplante de Médula Ósea/inmunología , Médula Ósea/efectos de la radiación , Rayos gamma , Antígenos de Histocompatibilidad/análisis , Animales , Sangre/efectos de la radiación , Perros , Envejecimiento Eritrocítico/efectos de la radiación , Calor , Leucocitos Mononucleares/inmunología , Prueba de Cultivo Mixto de LinfocitosRESUMEN
Irradiation of stored red blood cells (RBC) is increasingly utilized for patients who are immunosuppressed or on chemotherapeutic regimens. With the growing demand for irradiated cellular blood products, there has been an increasing need for transfusion services to store previously irradiated blood until needed for transfusion. The effect of irradiation on aging stored RBC has not been studied to date. Five units each of group A, RBC collected in CPD-Adsol (AS-1) with a prior shelf-life of 10, 20, 30, and 40 days, respectively, were divided equally utilizing a sterile docking device and stored at 1 to 6 degrees C. Baseline samples from each bag were obtained for the measurement of extracellular potassium (K+), plasma free hemoglobin (PFH), total lactate dehydrogenase (LD), and erythrocyte 2,3-DPG activity. One of each pair received 2,000 rads of gamma irradiation. Samples were obtained at 3 and 7 days post-irradiation, and multiples of 7 days until expiration. All irradiated units reached a state of K+ equilibrium at 60 to 70 mmol per L irrespective of the length of previous storage with an inverse relationship of RBC age at irradiation and the time required to reach the state of equilibrium. Increased K+ leakage from irradiated aging RBC suggests the need for including in vivo studies of cell survival to establish a post-irradiation storage life. Length of storage prior to irradiation had no effect on PFH, LD activity, and 2,3-diphosphoglycerate (2,3-DPG) activity compared to paired controls.
Asunto(s)
Envejecimiento Eritrocítico/efectos de la radiación , Eritrocitos/efectos de la radiación , 2,3-Difosfoglicerato , Conservación de la Sangre , Ácidos Difosfoglicéricos/sangre , Eritrocitos/química , Eritrocitos/enzimología , Rayos gamma , Hemoglobinas/metabolismo , Humanos , L-Lactato Deshidrogenasa/sangre , Potasio/sangre , Factores de TiempoRESUMEN
Red blood cells (pRBC) collected in citrate, phosphate, dextrose, adenine-formula 1 (CPDA-1) and citrate, phosphate, dextrose-adenine, manitol saline solution (CPD-ADSOL [AS-1]) anticoagulants are increasingly being stored for variable periods in transfusion service inventories following irradiation. While anecdotal reports of increased K+ following irradiation and storage have recently appeared in the literature, concomitant in vitro biochemical changes resulting from differences in anticoagulants have not been reported. Utilizing two venipunctures, two units each of 225 mL of blood from five volunteers were collected in anticoagulant-adjusted CPDA-1 and AS-1 bags. Within two hours of collection, each unit was equally divided. One of each pair was irradiated (2000 rads). Samples were analyzed on Days 0, 1, 3, 7, and every seven days to expiration. Irradiation resulted in a 2.3 fold increase in K+ during the first seven days of storage for both anticoagulants, although significantly greater K+ levels were observed in the CPDA-1 pairs compared to the AS-1 pairs. Comparison of glucose utilization, plasma free hemoglobin, 2,3-diphosphoglycerate (2,3-DPG) and lactate dehydrogenase between control and irradiated CPDA-1 and AS-1 pairs and between anticoagulants were documented.
Asunto(s)
Adenina , Anticoagulantes , Conservación de la Sangre , Citratos , Eritrocitos/efectos de la radiación , Glucosa , Manitol , Fosfatos , Cloruro de Sodio , 2,3-Difosfoglicerato , Glucemia/efectos de los fármacos , Glucemia/efectos de la radiación , Ácidos Difosfoglicéricos/sangre , Envejecimiento Eritrocítico/efectos de los fármacos , Envejecimiento Eritrocítico/efectos de la radiación , Eritrocitos/efectos de los fármacos , Eritrocitos/enzimología , Rayos gamma , Hemoglobinas/efectos de los fármacos , Hemoglobinas/efectos de la radiación , Humanos , L-Lactato Deshidrogenasa/sangre , MasculinoAsunto(s)
Eritrocitos/efectos de la radiación , Microondas , Oxígeno/sangre , Oxihemoglobinas/efectos de la radiación , Adaptación Fisiológica/efectos de la radiación , Animales , Transporte Biológico/fisiología , Transporte Biológico/efectos de la radiación , Envejecimiento Eritrocítico/fisiología , Envejecimiento Eritrocítico/efectos de la radiación , Eritrocitos/fisiología , Hemólisis/fisiología , Hemólisis/efectos de la radiación , Masculino , Oxígeno/farmacocinética , Oxígeno/efectos de la radiación , Oxihemoglobinas/metabolismo , Ratas , Factores de TiempoRESUMEN
Irradiated hamsters were infected with Leptospira interrogans serovar ballum in order to study the effects of impairment of the reticulo-macrophage system on the progression of the disease and the development of haemoglobinaemia. Infected and irradiated hamsters were compared with infected controls, irradiated controls and untreated controls. Red blood cells (RBCs) from untreated controls were biconcave disks while those from irradiated controls were echinocytes. Red blood cells from animals of both infected groups which were severely affected and moribund were in the form of pitted spherocytes and were associated with increased icteric indices and haemoglobinaemia. Red blood cells from less severely affected animals were echinocytic. Pitted spherocytes, however, were seen in some irradiated-infected hamsters before increased icteric indices and haemoglobinaemia were present. Both irradiated-infected and irradiated hamsters showed severe depletion of haemopoietic cells and lymphoid cells, with regeneration occurring in those animals which survived for longer. Red cell sequestration and erythrophagocytosis were present in infected-only hamsters. In irradiated-infected animals, severe anaemia was caused by both persistence of fixed macrophages, RBC sequestration and erythrophagocytosis, and haemorrhage due to vascular damage and renal papillary necrosis. A severe leucopaenia affected the irradiated hamsters' ability to suppress leptospiral multiplication resulting in the presence of larger numbers of organisms.
Asunto(s)
Envejecimiento Eritrocítico/efectos de la radiación , Hematopoyesis/efectos de la radiación , Enfermedad de Weil/sangre , Animales , Médula Ósea/patología , Cricetinae , Eritrocitos/ultraestructura , Rayos gamma , Hemoglobinas/análisis , Riñón/patología , Hígado/patología , Mesocricetus , Microscopía Electrónica de Rastreo , Bazo/patología , Enfermedad de Weil/patologíaAsunto(s)
Acetilcolinesterasa/efectos de la radiación , Membrana Eritrocítica/efectos de la radiación , Eritrocitos/efectos de la radiación , Animales , Tampones (Química) , Activación Enzimática/efectos de la radiación , Envejecimiento Eritrocítico/efectos de la radiación , Membrana Eritrocítica/enzimología , Rayos gamma , Técnicas In Vitro , Concentración Osmolar , RatasAsunto(s)
Envejecimiento Eritrocítico/efectos de la radiación , Deficiencia de Glucosafosfato Deshidrogenasa/sangre , Eritrocitos/efectos de la radiación , Deficiencia de Glucosafosfato Deshidrogenasa/radioterapia , Glutatión/sangre , Cuerpos de Heinz/metabolismo , Hemólisis , Humanos , Masculino , Metahemoglobina/metabolismo , Oxígeno , Radioterapia/efectos adversos , Rayos XRESUMEN
The immunosuppressive effect of extracorporeal irradiation of the blood (ECIB) in renal allotransplantation (RAT) has been investigated in a controlled clinical trial. A series of 101 uremic patients undergoing intermittent hemodialysis in preparation for RAT were allocated randomly to two groups; a treated group +ECIB (n=55), and a control group 0-ECIB (n= 46). ECIB treatment was given in the form of a fixed dose, with a transit dose of 400 rad and a cumulative dose of approximately 50 000 rad, both before and after RAT. A selective long-term lymphocytopenia was usually obtained, with no effect on immunoglobulins, complements or plasma proteins. Erythrocyte count was reduced in the majority of cases, but this had no limiting effect on the implementation of treatment. The shortest survival times and most deaths occurred in the +ECIB group. The first rejection episode occurred later in the +ECIB group, but there was no difference between the groups in the number of rejections during the first two months or during the whole observation period. There were more cases with two or more rejection episodes in the +ECIB than in the 0-ECIB group. There was no difference in graft survival, but shortest patient survival and most deaths after RAT occurred in the +ECIB group. It must therefore be concluded that the immunosuppressive effect of ECIB has no practical clinical relevance in RAT, and that ECIB given in the manner described involves the possibility of side-effects so serious that the method cannot be recommended.
Asunto(s)
Terapia de Inmunosupresión , Trasplante de Riñón , Efectos de la Radiación , Adolescente , Adulto , Anciano , Ensayos Clínicos como Asunto , Envejecimiento Eritrocítico/efectos de la radiación , Femenino , Rechazo de Injerto , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Trasplante HomólogoRESUMEN
Exposure to low levels of x-irradiation (50R) followed by phlebotomy (50% blood volume) one month post-irradiation, resulted in identifiable alterations of the erythropoietic status of the rat. The red cell indices revealed a decrease in mean corpuscular volume and an increase in mean corpuscular hemoglobin concentration. The half-life of 51Cr-labeled erythrocytes was 16.7 days versus 14.4 days for the untreated controls. These subjects also demonstrated hyperplastic marrows with an approximate 60% mean increase in marrow cellularity. A reproducible mortality of 25% was seen at 17 weeks post-irradiation; the one group of animals followed for an extended period exhibited an 86% cumulative mortality at 23 weeks. The observations supported the concept that x-irradiated rats exposed to a relatively low dose of X-rays (less than 10% LD/50) maintain a latent or residual injury of the bone marrow. These animals, when subsequently challenged by phlebotomy, are placed at greater risk with respect to their ability to survive.