From www.bloodjournal.org by guest on September 11, 2016. For personal use only.
7. Barbeau B, Mesnard J-M. Does the HBZ gene represent a new potential target for the treatment of adult T-cell leukemia? Int Rev Immunol. 2007;26(5-6):283-304.
(Tax-Tg) mouse model of adult T-cell leukemia/ lymphoma. Blood. 2009;114(13):2709-2720.
8. El Hajj H, El-Sabban M, Hasegawa H, et al. Therapyinduced selective loss of leukemia-initiating activity in murine adult T cell leukemia. J Exp Med. 2010;207(13):2785-2792.
10. Hishizawa M, Kanda J, Utsunomiya A, et al. Transplantation of allogeneic hematopoietic stem cells for adult T-cell leukemia: a nationwide retrospective study. Blood. 2010;116(8):1369-1376.
9. Yamazaki J, Mizukami T, Takizawa K, et al. Identification of cancer stem cells in a Tax-transgenic
© 2014 by The American Society of Hematology
l l l THROMBOSIS & HEMOSTASIS
Comment on Couturaud et al, page 2124
VTE risk and family history: provocative findings ----------------------------------------------------------------------------------------------------David A. Garcia
UNIVERSITY OF WASHINGTON
In this issue of Blood, Couturaud and colleagues take us one step closer to identifying a root cause for unprovoked venous thromboembolism (VTE) by comparing the VTE risk in first-degree relatives of patients who had experienced unprovoked venous thrombosis to the corresponding risk in first-degree relatives of patients whose thrombotic event was provoked by an environmental factor such as surgery.1
V
TE is a disease with many causes, and many environmental factors such as cancer, surgery, and estrogen use have been independently associated with an increased risk of venous thrombosis.2 Although the genetic contribution to VTE risk is illustrated by families deficient in endogenous anticoagulant proteins such as protein C, protein S, and antithrombin,3,4 our incomplete understanding of this disease is highlighted by the fact that many patients with apparently unprovoked VTE have no identifiable thrombophilia (inherited or acquired). It is not surprising that Couturaud et al1 found that first-degree relatives of VTE patients are themselves at increased risk to experience venous thrombosis. However, the observations that unprovoked (vs provoked) VTE and younger age at the time of first VTE are both associated with a higher risk of venous thrombosis in a first-degree relative have some important implications. One hypothesis that emerges from the findings of Couturaud et al is that patients with unprovoked VTE and a first-degree relative who had unprovoked VTE at a young age may be at higher risk for recurrent VTE than are otherwise similar patients. If confirmed in a prospective study, this sort of association could have implications for the recommended duration of anticoagulant therapy. Second, as the authors point out, the knowledge gained from this study may lead
clinicians to recommend that patients with a firstdegree relative who has experienced VTE avoid estrogen-containing contraceptive strategies and use aggressive VTE prophylaxis after surgery or hospitalization. Whether these common-sense suggestions should differ according to whether the first-degree relative’s clot was provoked or occurred at an older age is not clear, but the finding that a family history of VTE, even if provoked, increases risk is important. Since the only familial thrombophilia testing routinely performed in the Couturaud
study was for factor V Leiden and prothrombin 20210A gene variants, some of the increased risk observed among these first-degree relatives of VTE patients is likely attributable to known, inherited thrombophilias such as deficiency of protein C, protein S, or antithrombin. However, because these congenital deficiencies are quite rare, even within VTE patient cohorts, the associations described in the study by Couturaud et al strongly suggest that additional as yet unidentified genetic or epigenetic factors contribute to the development of VTE. Like many pathologic conditions, venous thrombosis is almost always the result of more than one contributing factor. The study by Couturaud and colleagues suggests that the search for additional inherited thrombophilic variations in the human genome should continue. Conflict-of-interest disclosure: The author declares no competing financial interests. n REFERENCES 1. Couturaud F, Leroyer C, Tromeur C, et al. Factors that predict thrombosis in relatives of patients with venous thromboembolism. Blood. 2014;124(13):2124-2130. 2. Rosendaal FR. Venous thrombosis: a multicausal disease. Lancet. 1999;353(9159):1167-1173. 3. Demers C, Ginsberg JS, Hirsh J, Henderson P, Blajchman MA. Thrombosis in antithrombin-III-deficient persons. Report of a large kindred and literature review. Ann Intern Med. 1992;116(9):754-761. 4. Lijfering WM, Brouwer JL, Veeger NJ, et al. Selective testing for thrombophilia in patients with first venous thrombosis: results from a retrospective family cohort study on absolute thrombotic risk for currently known thrombophilic defects in 2479 relatives. Blood. 2009;113(21):5314-5322. © 2014 by The American Society of Hematology
l l l VASCULAR BIOLOGY
Comment on Sikora et al, page 2150
Intravascular hemolysis: the sacrifice of few… ----------------------------------------------------------------------------------------------------Serge L. Y. Thomas
CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE; UNIVERSIT´E PIERRE ET MARIE CURIE
In this issue of Blood, Sikora et al demonstrate unequivocally that when erythrocytes are submitted to shear stress and hypoxia, only hemolysis contributes to the release of adenosine triphosphate (ATP), suggesting erythrocyte sacrifice as a primary mechanism for in vivo local purinergic signaling and blood flow regulation.1
L
ong ago, August Krogh arrived to the concept that “in the normal resting muscle a comparatively small number of capillaries…
BLOOD, 25 SEPTEMBER 2014 x VOLUME 124, NUMBER 13
should be open, so as to allow the passage of blood while muscular work should cause the opening up of a larger number.”2 A century
2011
From www.bloodjournal.org by guest on September 11, 2016. For personal use only.
later, regulation of blood flow through capillaries and the role played by erythrocytes in this process are still in debate. The elegant approach of experimental physiology presented by Sikora et al constitutes a decisive breakthrough in the field. It has been well documented that mechanical deformation, decreased oxygen partial pressure, and reduced pH, such as occur in microvessels of exercising muscle, induce ATP release and that this release activates purinergic receptors of vascular endothelial cells, resulting in synthesis of nitric oxide and metabolites of arachidonic acid known as powerful relaxation factors of smooth muscle vasculature contributing to match oxygen delivery with local need.3 However, the actual mechanism underlying ATP release remained in debate and was frequently attributed to complex signaling pathways opening up cystic fibrosis transmembrane conductance regulator (CFTR) channels.3 Unfortunately, no evidence of CFTR protein was found,4 and no CFTR single channel electrical activity was recorded in the erythrocyte membrane. The major contribution of Sikora et al is to provide an alternative and more simple mechanism not involving channels of intact cells: ATP release results from intravascular hemolysis! This result is not so surprising if we consider that blood is not a homogenous population of cells but rather is composed of a wide range of cells displaying very different homeostatic states, among them the senescent cells that are in a prolytic5,6 state before removal from circulation and constitute an easily accessible pool of ATP for local release. The role played by hemolysis is not novel because hemolysis has always been considered a potential factor contributing to stimulated ATP release in most previous investigations, but because of inappropriate evaluation of its actual involvement, due to a lack of paired measurement of free hemoglobin and ATP, its contribution to ATP release has been underestimated or simply overlooked. The novelty is that for the first time, Sikora et al were able to show that all regions displaying ATP release perfectly matched sites where cells had lysed. The clever use of ATP luminometry and luminescence ATP imaging demonstrated very nicely that brief spikes of luminescence due to ATP release from lysing cells was the sole mechanism of ATP release in these experiments.
2012
The authors show that in response to stimuli that elevate cyclic adenosine monophosphate (cAMP), the ATP releases were indistinguishable from those evoked by an equivalent volume of the solvent dimethyl sulfoxide (DMSO) alone, which rules out a possible contribution of cAMP agonists in ATP release but interestingly shows an effect of DMSO on ATP release, which illustrates the deleterious effects of DMSO on cell homeostasis. What does all this mean? Obviously, as clearly stated by Sikora et al, because hemolysis also occurs in vivo in physiological situations, such ATP release mechanisms are powerful physiological tools of blood flow regulation. It means that every factor influencing erythrocyte membrane fragility will also contribute to modulation of ATP release via hemolysis, but it also means that in absence of regulation these factors could be the origin of pathologies. Altered erythrocyte fragility and hemolysis are observed in many situations such as in vitro experimental conditions, blood storage, senescence, and pathologies. In light of the present work, it appears that there is an urgent need to better understand the mechanisms underlying hemolysis, but first, more caution should be taken when processing with blood samples. As the authors state, from the moment of blood collection to final assessment, erythrocytes are exposed to various physical perturbations that unavoidably cause rupture and hemolysis of many cells. For instance, the use of filters for removing white cells before experiments causes massive hemolysis and deprives the investigator of invaluable information on prolytic red cells and on stimuli intended to trigger regulated ATP release. It is a matter of fact that we know very little about conditions prevailing in cells toward the final days in the circulation. Lew et al7 have shown that the progressively increased intracellular calcium together with erythrocyte dehydration would activate a nonselective cationic channel,5,6 eventually leading to an unexpected light-density terminal state. Renewed attention should be paid to this phenomenon, which was also observed in sickle cells,8 to evaluate the role played by cationic channels as a possible triggering factor of hemolysis. The findings of Sikora et al also shed new light on the crucial problems of blood storage and transfusion. Today, the blood
preservation standards for transfusion require that $75% of the cells are still in the circulation 24 hours after transfusion and that hemolysis remains ,1% at the end of the storage period, but the key mechanisms responsible for hemolysis and cell disappearance remain unknown, and we can suspect the subsequent massive release of ATP to play a major role in posttransfusion harmful clinical outcomes. The work of Sikora et al raises many fascinating basic questions such as the role played by calcium in modulation of osmotic fragility and the link between altered membrane function in erythrocyte disorders9 (such as hereditary spherocytosis, elliptocytosis, ovalocytosis, stomatocytosis, sickle cell disease, and thalassemias) and the constant observation of cation leaks in these diseases. The possible collateral effects of the other molecules released together with ATP (ie, carbonic anhydrase) and the possible role played by microvesicles10 in this hemolytic process also deserve further investigation. Conflict-of-interest disclosure: The author declares no competing financial interests. n REFERENCES 1. Sikora J, Orlov SN, Furuya K, Grygorczyk R. Hemolysis is a primary ATP-release mechanism in human erythrocytes. Blood. 2014;124(13):2150-2157. 2. Krogh A. The supply of oxygen to the tissues and the regulation of the capillary circulation. J Physiol. 1919;52(6): 457-474. 3. Sprague RS, Stephenson AH, Ellsworth ML. Red not dead: signaling in and from erythrocytes. Trends Endocrinol Metab. 2007;18(9):350-355. 4. Hoffman JF, Dodson A, Wickrema A, Dib-Hajj SD. Tetrodotoxin-sensitive Na1 channels and muscarinic and purinergic receptors identified in human erythroid progenitor cells and red blood cell ghosts. Proc Natl Acad Sci USA. 2004;101(33):12370-12374. 5. Ponder E. The prolytic loss of K from human red cells. J Gen Physiol. 1947;30(3):235-246. 6. Crespo LM, Novak TS, Freedman JC. Calcium, cell shrinkage, and prolytic state of human red blood cells. Am J Physiol. 1987;252(2 Pt 1):C138-C152. 7. Lew VL, Daw N, Etzion Z, et al. Effects of age-dependent membrane transport changes on the homeostasis of senescent human red blood cells. Blood. 2007;110(4):1334-1342. 8. Bookchin RM, Etzion Z, Sorette M, Mohandas N, Skepper JN, Lew VL. Identification and characterization of a newly recognized population of high-Na1, low-K1, low-density sickle and normal red cells. Proc Natl Acad Sci USA. 2000;97(14):8045-8050. 9. Mohandas N, Gallagher PG. Red cell membrane: past, present, and future. Blood. 2008;112(10):3939-3948. 10. Alaarg A, Schiffelers RM, van Solinge WW, van Wijk R. Red blood cell vesiculation in hereditary hemolytic anemia. Front Physiol. 2013;4:article 365. doi:10.3389/fphys.2013.00365. © 2014 by The American Society of Hematology
BLOOD, 25 SEPTEMBER 2014 x VOLUME 124, NUMBER 13
From www.bloodjournal.org by guest on September 11, 2016. For personal use only.
2014 124: 2011-2012 doi:10.1182/blood-2014-08-595447
Intravascular hemolysis: the sacrifice of few… Serge L. Y. Thomas
Updated information and services can be found at: http://www.bloodjournal.org/content/124/13/2011.2.full.html Articles on similar topics can be found in the following Blood collections Free Research Articles (4041 articles) Information about reproducing this article in parts or in its entirety may be found online at: http://www.bloodjournal.org/site/misc/rights.xhtml#repub_requests Information about ordering reprints may be found online at: http://www.bloodjournal.org/site/misc/rights.xhtml#reprints Information about subscriptions and ASH membership may be found online at: http://www.bloodjournal.org/site/subscriptions/index.xhtml
Blood (print ISSN 0006-4971, online ISSN 1528-0020), is published weekly by the American Society of Hematology, 2021 L St, NW, Suite 900, Washington DC 20036. Copyright 2011 by The American Society of Hematology; all rights reserved.
7. Barbeau B, Mesnard J-M. Does the HBZ gene represent a new potential target for the treatment of adult T-cell leukemia? Int Rev Immunol. 2007;26(5-6):283-304.
(Tax-Tg) mouse model of adult T-cell leukemia/ lymphoma. Blood. 2009;114(13):2709-2720.
8. El Hajj H, El-Sabban M, Hasegawa H, et al. Therapyinduced selective loss of leukemia-initiating activity in murine adult T cell leukemia. J Exp Med. 2010;207(13):2785-2792.
10. Hishizawa M, Kanda J, Utsunomiya A, et al. Transplantation of allogeneic hematopoietic stem cells for adult T-cell leukemia: a nationwide retrospective study. Blood. 2010;116(8):1369-1376.
9. Yamazaki J, Mizukami T, Takizawa K, et al. Identification of cancer stem cells in a Tax-transgenic
© 2014 by The American Society of Hematology
l l l THROMBOSIS & HEMOSTASIS
Comment on Couturaud et al, page 2124
VTE risk and family history: provocative findings ----------------------------------------------------------------------------------------------------David A. Garcia
UNIVERSITY OF WASHINGTON
In this issue of Blood, Couturaud and colleagues take us one step closer to identifying a root cause for unprovoked venous thromboembolism (VTE) by comparing the VTE risk in first-degree relatives of patients who had experienced unprovoked venous thrombosis to the corresponding risk in first-degree relatives of patients whose thrombotic event was provoked by an environmental factor such as surgery.1
V
TE is a disease with many causes, and many environmental factors such as cancer, surgery, and estrogen use have been independently associated with an increased risk of venous thrombosis.2 Although the genetic contribution to VTE risk is illustrated by families deficient in endogenous anticoagulant proteins such as protein C, protein S, and antithrombin,3,4 our incomplete understanding of this disease is highlighted by the fact that many patients with apparently unprovoked VTE have no identifiable thrombophilia (inherited or acquired). It is not surprising that Couturaud et al1 found that first-degree relatives of VTE patients are themselves at increased risk to experience venous thrombosis. However, the observations that unprovoked (vs provoked) VTE and younger age at the time of first VTE are both associated with a higher risk of venous thrombosis in a first-degree relative have some important implications. One hypothesis that emerges from the findings of Couturaud et al is that patients with unprovoked VTE and a first-degree relative who had unprovoked VTE at a young age may be at higher risk for recurrent VTE than are otherwise similar patients. If confirmed in a prospective study, this sort of association could have implications for the recommended duration of anticoagulant therapy. Second, as the authors point out, the knowledge gained from this study may lead
clinicians to recommend that patients with a firstdegree relative who has experienced VTE avoid estrogen-containing contraceptive strategies and use aggressive VTE prophylaxis after surgery or hospitalization. Whether these common-sense suggestions should differ according to whether the first-degree relative’s clot was provoked or occurred at an older age is not clear, but the finding that a family history of VTE, even if provoked, increases risk is important. Since the only familial thrombophilia testing routinely performed in the Couturaud
study was for factor V Leiden and prothrombin 20210A gene variants, some of the increased risk observed among these first-degree relatives of VTE patients is likely attributable to known, inherited thrombophilias such as deficiency of protein C, protein S, or antithrombin. However, because these congenital deficiencies are quite rare, even within VTE patient cohorts, the associations described in the study by Couturaud et al strongly suggest that additional as yet unidentified genetic or epigenetic factors contribute to the development of VTE. Like many pathologic conditions, venous thrombosis is almost always the result of more than one contributing factor. The study by Couturaud and colleagues suggests that the search for additional inherited thrombophilic variations in the human genome should continue. Conflict-of-interest disclosure: The author declares no competing financial interests. n REFERENCES 1. Couturaud F, Leroyer C, Tromeur C, et al. Factors that predict thrombosis in relatives of patients with venous thromboembolism. Blood. 2014;124(13):2124-2130. 2. Rosendaal FR. Venous thrombosis: a multicausal disease. Lancet. 1999;353(9159):1167-1173. 3. Demers C, Ginsberg JS, Hirsh J, Henderson P, Blajchman MA. Thrombosis in antithrombin-III-deficient persons. Report of a large kindred and literature review. Ann Intern Med. 1992;116(9):754-761. 4. Lijfering WM, Brouwer JL, Veeger NJ, et al. Selective testing for thrombophilia in patients with first venous thrombosis: results from a retrospective family cohort study on absolute thrombotic risk for currently known thrombophilic defects in 2479 relatives. Blood. 2009;113(21):5314-5322. © 2014 by The American Society of Hematology
l l l VASCULAR BIOLOGY
Comment on Sikora et al, page 2150
Intravascular hemolysis: the sacrifice of few… ----------------------------------------------------------------------------------------------------Serge L. Y. Thomas
CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE; UNIVERSIT´E PIERRE ET MARIE CURIE
In this issue of Blood, Sikora et al demonstrate unequivocally that when erythrocytes are submitted to shear stress and hypoxia, only hemolysis contributes to the release of adenosine triphosphate (ATP), suggesting erythrocyte sacrifice as a primary mechanism for in vivo local purinergic signaling and blood flow regulation.1
L
ong ago, August Krogh arrived to the concept that “in the normal resting muscle a comparatively small number of capillaries…
BLOOD, 25 SEPTEMBER 2014 x VOLUME 124, NUMBER 13
should be open, so as to allow the passage of blood while muscular work should cause the opening up of a larger number.”2 A century
2011
From www.bloodjournal.org by guest on September 11, 2016. For personal use only.
later, regulation of blood flow through capillaries and the role played by erythrocytes in this process are still in debate. The elegant approach of experimental physiology presented by Sikora et al constitutes a decisive breakthrough in the field. It has been well documented that mechanical deformation, decreased oxygen partial pressure, and reduced pH, such as occur in microvessels of exercising muscle, induce ATP release and that this release activates purinergic receptors of vascular endothelial cells, resulting in synthesis of nitric oxide and metabolites of arachidonic acid known as powerful relaxation factors of smooth muscle vasculature contributing to match oxygen delivery with local need.3 However, the actual mechanism underlying ATP release remained in debate and was frequently attributed to complex signaling pathways opening up cystic fibrosis transmembrane conductance regulator (CFTR) channels.3 Unfortunately, no evidence of CFTR protein was found,4 and no CFTR single channel electrical activity was recorded in the erythrocyte membrane. The major contribution of Sikora et al is to provide an alternative and more simple mechanism not involving channels of intact cells: ATP release results from intravascular hemolysis! This result is not so surprising if we consider that blood is not a homogenous population of cells but rather is composed of a wide range of cells displaying very different homeostatic states, among them the senescent cells that are in a prolytic5,6 state before removal from circulation and constitute an easily accessible pool of ATP for local release. The role played by hemolysis is not novel because hemolysis has always been considered a potential factor contributing to stimulated ATP release in most previous investigations, but because of inappropriate evaluation of its actual involvement, due to a lack of paired measurement of free hemoglobin and ATP, its contribution to ATP release has been underestimated or simply overlooked. The novelty is that for the first time, Sikora et al were able to show that all regions displaying ATP release perfectly matched sites where cells had lysed. The clever use of ATP luminometry and luminescence ATP imaging demonstrated very nicely that brief spikes of luminescence due to ATP release from lysing cells was the sole mechanism of ATP release in these experiments.
2012
The authors show that in response to stimuli that elevate cyclic adenosine monophosphate (cAMP), the ATP releases were indistinguishable from those evoked by an equivalent volume of the solvent dimethyl sulfoxide (DMSO) alone, which rules out a possible contribution of cAMP agonists in ATP release but interestingly shows an effect of DMSO on ATP release, which illustrates the deleterious effects of DMSO on cell homeostasis. What does all this mean? Obviously, as clearly stated by Sikora et al, because hemolysis also occurs in vivo in physiological situations, such ATP release mechanisms are powerful physiological tools of blood flow regulation. It means that every factor influencing erythrocyte membrane fragility will also contribute to modulation of ATP release via hemolysis, but it also means that in absence of regulation these factors could be the origin of pathologies. Altered erythrocyte fragility and hemolysis are observed in many situations such as in vitro experimental conditions, blood storage, senescence, and pathologies. In light of the present work, it appears that there is an urgent need to better understand the mechanisms underlying hemolysis, but first, more caution should be taken when processing with blood samples. As the authors state, from the moment of blood collection to final assessment, erythrocytes are exposed to various physical perturbations that unavoidably cause rupture and hemolysis of many cells. For instance, the use of filters for removing white cells before experiments causes massive hemolysis and deprives the investigator of invaluable information on prolytic red cells and on stimuli intended to trigger regulated ATP release. It is a matter of fact that we know very little about conditions prevailing in cells toward the final days in the circulation. Lew et al7 have shown that the progressively increased intracellular calcium together with erythrocyte dehydration would activate a nonselective cationic channel,5,6 eventually leading to an unexpected light-density terminal state. Renewed attention should be paid to this phenomenon, which was also observed in sickle cells,8 to evaluate the role played by cationic channels as a possible triggering factor of hemolysis. The findings of Sikora et al also shed new light on the crucial problems of blood storage and transfusion. Today, the blood
preservation standards for transfusion require that $75% of the cells are still in the circulation 24 hours after transfusion and that hemolysis remains ,1% at the end of the storage period, but the key mechanisms responsible for hemolysis and cell disappearance remain unknown, and we can suspect the subsequent massive release of ATP to play a major role in posttransfusion harmful clinical outcomes. The work of Sikora et al raises many fascinating basic questions such as the role played by calcium in modulation of osmotic fragility and the link between altered membrane function in erythrocyte disorders9 (such as hereditary spherocytosis, elliptocytosis, ovalocytosis, stomatocytosis, sickle cell disease, and thalassemias) and the constant observation of cation leaks in these diseases. The possible collateral effects of the other molecules released together with ATP (ie, carbonic anhydrase) and the possible role played by microvesicles10 in this hemolytic process also deserve further investigation. Conflict-of-interest disclosure: The author declares no competing financial interests. n REFERENCES 1. Sikora J, Orlov SN, Furuya K, Grygorczyk R. Hemolysis is a primary ATP-release mechanism in human erythrocytes. Blood. 2014;124(13):2150-2157. 2. Krogh A. The supply of oxygen to the tissues and the regulation of the capillary circulation. J Physiol. 1919;52(6): 457-474. 3. Sprague RS, Stephenson AH, Ellsworth ML. Red not dead: signaling in and from erythrocytes. Trends Endocrinol Metab. 2007;18(9):350-355. 4. Hoffman JF, Dodson A, Wickrema A, Dib-Hajj SD. Tetrodotoxin-sensitive Na1 channels and muscarinic and purinergic receptors identified in human erythroid progenitor cells and red blood cell ghosts. Proc Natl Acad Sci USA. 2004;101(33):12370-12374. 5. Ponder E. The prolytic loss of K from human red cells. J Gen Physiol. 1947;30(3):235-246. 6. Crespo LM, Novak TS, Freedman JC. Calcium, cell shrinkage, and prolytic state of human red blood cells. Am J Physiol. 1987;252(2 Pt 1):C138-C152. 7. Lew VL, Daw N, Etzion Z, et al. Effects of age-dependent membrane transport changes on the homeostasis of senescent human red blood cells. Blood. 2007;110(4):1334-1342. 8. Bookchin RM, Etzion Z, Sorette M, Mohandas N, Skepper JN, Lew VL. Identification and characterization of a newly recognized population of high-Na1, low-K1, low-density sickle and normal red cells. Proc Natl Acad Sci USA. 2000;97(14):8045-8050. 9. Mohandas N, Gallagher PG. Red cell membrane: past, present, and future. Blood. 2008;112(10):3939-3948. 10. Alaarg A, Schiffelers RM, van Solinge WW, van Wijk R. Red blood cell vesiculation in hereditary hemolytic anemia. Front Physiol. 2013;4:article 365. doi:10.3389/fphys.2013.00365. © 2014 by The American Society of Hematology
BLOOD, 25 SEPTEMBER 2014 x VOLUME 124, NUMBER 13
From www.bloodjournal.org by guest on September 11, 2016. For personal use only.
2014 124: 2011-2012 doi:10.1182/blood-2014-08-595447
Intravascular hemolysis: the sacrifice of few… Serge L. Y. Thomas
Updated information and services can be found at: http://www.bloodjournal.org/content/124/13/2011.2.full.html Articles on similar topics can be found in the following Blood collections Free Research Articles (4041 articles) Information about reproducing this article in parts or in its entirety may be found online at: http://www.bloodjournal.org/site/misc/rights.xhtml#repub_requests Information about ordering reprints may be found online at: http://www.bloodjournal.org/site/misc/rights.xhtml#reprints Information about subscriptions and ASH membership may be found online at: http://www.bloodjournal.org/site/subscriptions/index.xhtml
Blood (print ISSN 0006-4971, online ISSN 1528-0020), is published weekly by the American Society of Hematology, 2021 L St, NW, Suite 900, Washington DC 20036. Copyright 2011 by The American Society of Hematology; all rights reserved.
