Methods and treatment of trauma

Abstract

To provide oxygen reduced stored blood for use in transfusion therapy for a hemorrhagic trauma subject in need of transfusion.SOLUTION: The present invention provides oxygen reduced stored blood having an initial oxygen saturation (SO2) level of 20% or less and maintained at an SO2 level of 20% or less during a storage period.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to U.S. Provisional Application No. 62 / 508,783, filed on May 19, 2017, the entire content of which is incorporated herein by reference.

[0002] Government Rights This invention was made with government support under R44HL132172, awarded by the National Heart, Lung, and Blood Institute. The government has certain rights in this invention.

[0003] Field of the Invention This disclosure relates to the treatment of trauma and hemorrhagic shock.

Background Art

[0004] There were 5.1 million deaths due to injury in 2010, which exceeded the combined number of deaths due to HIV, tuberculosis, and malaria (3.8 million). See Norton, et al., “Global Health Injuries” (The NEJM, 368:1723-30 (2013)) (hereinafter “Norton 2013”) (which is hereby incorporated by reference in its entirety). Injuries include unintentional injuries (e.g., traffic accidents, falls, and burns), as well as intentional injuries (e.g., self-harm, interpersonal violence, war, and conflict). See Norton 2013. The number of deaths due to injury increased by 24% worldwide between 1990 and 2010 and by 23% in the United States between 2000 and 2010. See Norton 2013. In addition, at least 20% of all trauma deaths are the result of injuries that could have been survivable, and thus such trauma deaths are preventable with optimal care. See Fox et al., “Earlier Endpoints are Required for Hemorrhagic Shock Trials Among Severely Injured Patients.” Shock, 47:567-73 (2017) (which is hereby incorporated by reference in its entirety). Given the percentage of preventable deaths, there is an urgent need to develop therapies for avoidable complications that lead to death.

[0005] Penetrating trauma (e.g., gunshot or stab wounds) and blunt trauma (e.g., trauma from falls or motor vehicle accidents) are the main causes of hemorrhagic trauma. The resulting shock is a state in which oxygen supply to tissues is insufficient due to massive bleeding, which causes oxygen debt, anaerobic metabolism, and an increase in plasma lactate levels. If shock cannot be reversed by restoring circulation and oxygen delivery, permanent tissue damage, multiple organ failure, and death can result.

[0006] Clinical sequelae of hemorrhagic trauma and shock include death due to blood loss within hours of trauma, and death after 24 hours from the pathological conditions caused by trauma and massive transfusion. Such pathological conditions include multiple organ failure (including acute traumatic coagulopathy or failure of the lungs, kidneys, and liver from inflammation), and infection / sepsis from transfusion-related immunomodulation. All pathological conditions are enhanced the lower the quality of the blood products transfused and the greater the amount of pRBCs transfused.

[0007] One approach to treating hemorrhagic shock is the use of crystalloids for resuscitation. However, the use of crystalloids causes an increase in pathological conditions and death by inducing trauma-induced coagulopathy. For at least this reason, early administration of blood components to reverse shock caused by hemorrhagic trauma has been advocated. Packed red blood cells (pRBCs) are transfused to hemorrhagic trauma patients for the recovery of the lost blood volume, the recovery of oxygen-carrying capacity in the patient, and the recovery of oxidative metabolism in tissues from anaerobic metabolism. However, the use of pRBCs is not without the risk of complications including antigen mismatching, pathogen transmission, circulatory overload, and degradation of pRBCs during ex vivo storage.

[0008] Stored blood undergoes progressive deterioration associated with various storage lesions (including in particular hemolysis, hemoglobin degradation, and reduction in ATP and 2,3-DPG concentrations) when stored conventionally. The effects of this progressive deterioration during storage appear, for example, as a reduction in the in vivo recovery rate at 24 hours when transfused to patients. A rapid decrease in hematocrit due to the reduction in the 24-hour recovery rate can, in severe cases, result in a delayed hemolytic transfusion reaction (DHTR). Other complications, such as the systemic inflammatory response syndrome (SIRS), transfusion-related acute lung injury (TRALI), and transfusion-related immunomodulation (TRIM), are associated with the transfusion of stored blood, but the underlying causes remain unclear.

[0009] Stored RBCs show lower quality (e.g., increased RBC fraction removed, decreased oxygen exchange capacity, reduced deformability) and increased toxicity, often manifested as clinical sequelae of transfusion therapy, even when transfused within the current 6-week limit. There are numerous papers in the literature supporting this view and the number is increasing. See: Zimring, “Established and theoretical factors to consider in assessing the red cell storage lesion,” Blood, 125:2185-90 (2015); Zhu et al., “Impaired adenosine-5’-triphosphate release from red blood cells promotes their adhesion to endothelial cells: a mechanism of hypoxemia after transfusion,” Critical care medicine, 39:2478-86 (2011); Weinberg et al., “Red blood cell age and potentiation of transfusion-related pathology in trauma patients,” Transfusion, 51:867-73 (2011); Spinella et al., “Does the storage duration of blood products affect outcomes in critically ill patients?” Transfusion 51:1644-50 (2011); Roback et al., “Insufficient nitric oxide bioavailability: a hypothesis to explain adverse effects of red blood cell transfusion,” Transfusion, 51:859-66 (2011); Reynolds et al.,"The transfusion problem: role of aberrant S-nitrosylation," Transfusion, 51:852 - 8 (2011); Kim-Shapiro et al., "Storage lesion: role of red blood cell breakdown," Transfusion, 51:844 - 51 (2011); Jy et al., "Microparticles in stored red blood cells as potential mediators of transfusion complications," Transfusion, 51:886 - 93 (2011); Hod et al., "Transfusion of human volunteers with older, stored red blood cells produces extravascular hemolysis and circulating non-transferrin-bound iron," Blood, 118:6675 - 82 (2011); Flegel et al., "Does prolonged storage of red blood cells cause harm?" British journal of haematology 165:3 - 16 (2014); Redlin et al., "Red blood cell storage duration is associated with various clinical outcomes in pediatric cardiac surgery," Transfusion medicine and hemotherapy: offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie 41:146 - 51 (2014); Rogers et al.,"Storage duration of red blood cell transfusion and Clostridium difficile infection: a within person comparison," PLoS One 9:e89332(2014); Spinella et al., "Properties of stored red blood cells: understanding immune and vascular reactivity," Transfusion 51:894 - 900(2011); Brown et al., "Length of red cell unit storage and risk for delirium after cardiac surgery," Anesth Analg,119:242 - 50(2014); Wang et al., "Transfusion of older stored blood worsens outcomes in canines depending on the presence and severity of pneumonia," Transfusion,54:1712 - 24(2014); Liu et al., "Mechanism of faster NO scavenging by older stored red blood cells," Redox biology,2:211 - 9(2014); Prestia et al., "Transfusion of stored blood impairs host defenses against Gram - negative pathogens in mice," Transfusion 54:2842 - 51(2014); D’Alessandro et al.,"An update on red blood cell storage lesions, as gleaned through biochemistry and omics technologies," Transfusion, 55:205-19 (2015) (these are all incorporated herein by reference). Extensive in vitro studies have clearly shown the breakdown of conventional RBCs during storage (storage lesions). A series of emerging metabolome studies have shown the occurrence of storage lesions at the molecular level. See the following: Roback et al., "Metabolomics of AS-1 RBCs Storage," Transfusion medicine reviews (2014); D'Alessandro et al., "Metabolomics of AS-5 RBCs supernatants following routine storage," Vox sanguinis (2014); D'Alessandro et al., "Routine storage of red blood cell (RBC) units in additive solution-3: a comprehensive investigation of the RBC metabolome," Transfusion 55:1155-68 (2015); D'Alessandro et al., "Red blood cell storage in additive solution-7 preserves energy and redox metabolism: a metabolomics approach," Transfusion (2015); Wither et al., "Hemoglobin oxidation at functional amino acid residues during routine storage of red blood cells," Transfusion (2015); D'Alessandro et al.,"Citrate metabolism in red blood cells stored in additive solution-3," Transfusion (2016); D'Alessandro et al., "Omics markers of the red cell storage lesion and metabolic linkage," Blood Transfus, 15:137-44 (2017) (the entireties of these are incorporated herein by reference). There is a need to reduce or prevent this degradation to increase the effectiveness of transfusion (delivering more O2 to peripheral tissues than immediately after transfusion) and to reduce death due to hemorrhagic trauma.

[0010] Acidification damage causes many RBC storage alterations and downstream events in conventional stored blood, so a method is needed to reduce the degree of oxidative stress and thus reduce RBC storage alterations. A number of approaches have been developed with the aim of minimizing storage alterations and improving transfusion outcomes. Approaches include additive solutions (e.g., U.S. Patent No. 4,769,318 to Hamasaki et al., and U.S. Patent No. 4,880,786 to Sasakawa et al., U.S. Patent No. 6,447,987 to Hess et al.), cryopreservation (see U.S. Patent No. 6,413,713 to Serebrennikov, Chaplin et al., “Blood Cells for Transfusion,” Blood, 59:1118-20 (1982), and Valeri et al., “The survival, function, and hemolysis of human RBCs stored at 4 degrees C in additive solution (AS-1, AS-3, or AS-5) for 42 days and then biochemically modified, frozen, thawed, washed, and stored at 4 degrees C in sodium chloride and glucose solution for 24 hours,” Transfusion, 40:1341-5 (2000)) (all of which are hereby incorporated by reference in their entirety).

[0011] One approach that has been found to successfully improve blood quality and expand its practicality is through storage under oxygen depletion and anaerobic conditions. Advantages of storing blood under oxygen depletion conditions include improved levels of ATP and 2,3-DPG, as well as reduced hemolysis. U.S. Patent No. 5,624,794 to Bitensky et al., U.S. Patent No. 6,162,396 to Bitensky et al., and U.S. Patent No. 5,476,764 to Bitensky (all of which are incorporated herein by reference in their entirety) are directed to the storage of red blood cells under oxygen depletion conditions. U.S. Patent No. 5,789,151 to Bitensky et al. is directed to blood storage additive solutions (which is incorporated herein by reference in its entirety). U.S. Patent No. 6,162,396 ('396 patent) to Bitensky et al. (which is incorporated herein by reference in its entirety) discloses an anaerobic storage bag for blood storage, having an oxygen-impermeable outer layer, a red blood cell (RBC)-compatible inner layer that is permeable to oxygen, and an oxygen scrubber disposed between the inner and outer layers.

[0012] Moreover, the storage of blood under oxygen depletion conditions can also result in a reduction in microparticle levels, a reduction in loss of deformability, a reduction in oxidation of lipids and proteins, and an increase in post-transfusion survival rate, compared to blood stored under conventional conditions. See Yoshida et al., “The effects of additive solution pH and metabolic rejuvenation on anaerobic storage of red cells,” Transfusion 48:2096-2105(2008) and Yoshida, T., et al. “Reduction of microparticle generation during anaerobic storage of red blood cells. Transfusion”, 52, 83A(2012) (the entireties of which are incorporated herein by reference). Anaerobic stored RBCs further result in an increase in the in vivo recovery rate at 24 hours after autologous transfusion, an increase in 2,3-DPG and ATP levels, a decrease in hemolysis, and beneficial remodeling in the metabolic pathway. See Reisz et al. “Oxidative modifications of glyceraldehyde 3-phosphate dehydrogenase regulate metabolic reprogramming of stored red blood cells,” Blood, 128:e32-42(2016); and Yoshida et al., “Extended storage of red blood cells under anaerobic conditions,” Vox sanguinis 92:22-31(2007) (the entireties of which are incorporated herein by reference).

[0013] In the present disclosure, the inventors demonstrate that, as previously demonstrated using human blood, oxygen reduction (OR) or oxygen and carbon dioxide reduction (OCR) blood from rats results in improvements in ATP and 2,3-DPG during storage compared to conventional stored blood. Thus, OR or OCR rat RBCs are predicted to similarly reduce microparticles, improve deformability, reduce lipid and protein oxidation, and increase post-transfusion survival.

[0014] Herein, the inventors demonstrate for the first time in rats that OR and OCR blood provides surprising improvements in clinical outcomes when transfused to treat hemorrhagic trauma. The inventors used a rat hemorrhagic shock resuscitation model to show that OR or OCR RBCs provide a reduction in organ injury compared to conventional stored blood. In addition, OR or OCR RBCs provide the restoration of the shock state using a lower volume of pRBCs. Finally, OR or OCR RBCs more rapidly stabilized hemodynamics compared to conventional stored pRBCs of the same storage period when transfused to treat hemorrhagic shock.

[0015] OR and OCR RBCs provide an improved method for the treatment of trauma resulting in blood loss, reducing mortality and morbidity compared to conventional stored blood. OR and OCR RBCs provide a reduction in organ failure, including reduction in marker levels of lung and liver injury. OR and OCR RBCs further provide a reduction in the amount of blood required for the restoration and stabilization of hemodynamic function. Thus, OR and OCR RBCs may provide a reduction in the amount of RBCs required for transfusion therapy when treating hemorrhagic trauma. The quality improvements of OR and OCR also provide an unexpected reduction in trauma-related organ injury, morbidity, and mortality in addition to the previously demonstrated improvement in oxygen delivery capacity in stored RBCs. SUMMARY OF THE INVENTION

[0016] The present disclosure provides a method for treating low mean arterial pressure in a subject in need of treatment for low mean arterial pressure, the method including supplying oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, wherein after supplying the oxygen-reduced blood to the subject in need of treatment, the mean arterial pressure in the subject in need of treatment increases and the low mean arterial pressure is due to hemorrhagic trauma, and including this.

[0017] The present disclosure provides a method for reducing the amount of blood required for transfusion in a trauma patient in need of transfusion, the method including supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage, and including this.

[0018] The present disclosure provides a method for reducing hemorrhagic shock in a trauma patient in need of reduction of hemorrhagic shock, the method including supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage, wherein the trauma patient has a lactate level between 0.5 and 2.5 millimoles per liter (mmol / L) per liter before the supply and the hemorrhagic shock returns to normal, and including this.

[0019] The present disclosure provides a method for reducing liver injury in a trauma patient in need of transfusion therapy, the method including supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage, and including this.

[0020] The present disclosure provides a method for reducing renal failure in a hemorrhagic trauma patient in need of transfusion therapy, the method including supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage, and including this.

[0021] The present disclosure provides a method for reducing lung injury in a hemorrhagic trauma patient in need of transfusion therapy, the method including supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage, and including this.

[0022] The present disclosure is provided with reference to the accompanying drawings.

Brief Description of the Drawings

[0023]

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Mode for Carrying Out the Invention

[0024] The examples shown in this specification illustrate some embodiments in the present disclosure, but should not be construed as limiting the scope of the present disclosure in any way.

[0025] The method of the present disclosure provides, and includes, supplying a hemorrhagic trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. The method also provides supplying a hemorrhagic trauma patient with oxygen-reduced stored blood having an oxygen saturation between 15% and 20% before and during storage. The method also provides supplying a hemorrhagic trauma patient with oxygen-reduced stored blood having an oxygen saturation between 10% and 15% before and during storage. The method also provides supplying a hemorrhagic trauma patient with oxygen-reduced stored blood having an oxygen saturation between 5% and 10% before and during storage. The method also provides supplying a hemorrhagic trauma patient with oxygen-reduced stored blood having an oxygen saturation between 3% and 5% before and during storage.

[0026] The method also provides supplying a person with hemorrhagic shock with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during transfusion. The method also provides supplying a person with hemorrhagic trauma with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during transfusion. The method also includes a method that includes transfusing a patient with an increased risk of trauma due to surgery with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. The method of supplying oxygen-reduced stored blood having an initial oxygen saturation of 20% or less includes supplying oxygen-reduced stored blood having an initial oxygen saturation of 10% or less. The method of supplying oxygen-reduced stored blood having an initial oxygen saturation of 20% or less further includes supplying oxygen-reduced stored blood having an initial oxygen saturation of 5% or less. The method of supplying oxygen-reduced stored blood having an initial oxygen saturation of 20% or less further includes supplying oxygen-reduced stored blood having an initial oxygen saturation of 3% or less.

[0027] The method of the present disclosure provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation of 20% or less before and during storage for a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks, and includes this. The method also provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation of 15% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation of 10% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method further provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation of 5% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method further provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation of 3% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation between 3% and 5% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation between 5% and 10% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks.The method also provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation between 10% and 15% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides for supplying oxygen-reduced stored blood for the treatment of trauma, wherein the oxygen-reduced stored blood has an oxygen saturation between 15% and 20% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks.

[0028] The method of the present disclosure provides for supplying an oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to a trauma patient, and includes this. In one aspect, the trauma patient suffers from a head trauma, a penetrating wound, a blunt trauma, an injury due to a fall, or an injury due to a motor vehicle accident. In another aspect, the trauma patient is a hemorrhagic trauma patient. In yet another aspect, the trauma patient is bleeding due to surgery, a penetrating wound, a blunt trauma, an injury due to a fall, or an injury due to a motor vehicle accident.

[0029] In one aspect of the present disclosure, the trauma patient or the hemorrhagic trauma patient is a subject in need of OR and OCR stored blood. In an aspect of the present disclosure, the trauma patient is a hemorrhagic trauma patient in need of one or more units of blood as a transfusion therapy. In an aspect of the present disclosure, the trauma patient is a hemorrhagic trauma patient in need of two or more units of blood as a transfusion therapy. In an aspect of the present disclosure, the trauma patient is a hemorrhagic trauma patient in need of three or more units of blood as a transfusion therapy.

[0030] In one aspect of the present disclosure, a trauma patient is a patient in a hemorrhagic shock state. In one aspect, the trauma patient is in a hemorrhagic shock state due to head trauma, penetrating trauma, blunt trauma, injury from a fall, or injury from a motor vehicle accident. In an aspect of the present disclosure, the hemorrhagic trauma patient is a patient with Class I hemorrhage. In another aspect, the hemorrhagic trauma patient is a patient with Class II hemorrhage. In another aspect, the hemorrhagic trauma patient is a patient with Class III hemorrhage. In another aspect, the hemorrhagic trauma patient is a patient with Class IV hemorrhage. In one aspect of the present disclosure, the hemorrhagic trauma patient loses up to 15% of the blood volume. In another aspect, the hemorrhagic trauma patient loses between 15% and 30% of the blood volume. In another aspect, the hemorrhagic trauma patient loses between 30% and 40% of the blood volume. In another aspect, the hemorrhagic trauma patient loses more than 40% of the blood volume.

[0031] The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits one or more signs selected from the group consisting of a decrease in mean arterial pressure, a decrease in hematocrit, an increase in lactate, an increase in glucose, an increase in aspartate aminotransferase (AST), an increase in alanine aminotransferase (ALT), an increase in urinary neutrophil gelatinase-associated lipocalin (u-NGAL), an increase in serum creatinine, and an increase in blood urea nitrogen, and includes the same. In one aspect of the present disclosure, the patient in need of transfusion therapy using OR or OCR RBCs is a hemorrhagic trauma patient with a decreased mean arterial pressure. The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits an increase in aspartate aminotransferase (AST) and an increase in alanine aminotransferase (ALT), and includes the same. The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits a decrease in mean arterial pressure and an increase in lactate, and includes the same. The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits an increase in aspartate aminotransferase (AST), an increase in alanine aminotransferase (ALT), and an increase in blood urea nitrogen, and includes the same. The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits an increase in aspartate aminotransferase (AST), an increase in alanine aminotransferase (ALT), an increase in serum creatinine, and an increase in blood urea nitrogen, and includes the same. The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits an increase in lactate and an increase in glucose, and includes the same. The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits an increase in urinary neutrophil gelatinase-associated lipocalin (u-NGAL), an increase in serum creatinine, and an increase in blood urea nitrogen, and includes the same.

[0032] In another aspect, patients in need of transfusion therapy using OR or OCR RBCs are hemorrhagic trauma patients with decreased hematocrit. In another aspect, patients in need of transfusion therapy using OR or OCR RBCs are hemorrhagic trauma patients with increased lactate. In yet another aspect, patients in need of transfusion therapy using OR or OCR RBCs are hemorrhagic trauma patients with increased glucose. In a further aspect, hemorrhagic trauma patients with increased aspartate aminotransferase (AST). In another aspect, patients in need of transfusion therapy using OR or OCR RBCs are hemorrhagic trauma patients with increased alanine aminotransferase (ALT). In another aspect, patients in need of transfusion therapy using OR or OCR RBCs are hemorrhagic trauma patients with increased urinary neutrophil gelatinase-associated lipocalin (u-NGAL). In another aspect, patients in need of transfusion therapy using OR or OCR RBCs are hemorrhagic trauma patients with increased serum creatinine. In another aspect, patients in need of transfusion therapy using OR or OCR RBCs are hemorrhagic trauma patients with increased blood urea nitrogen.

[0033] In one aspect of the present disclosure, OR and OCR stored blood for use in transfusion therapy of trauma patients in need of transfusion therapy has an initial oxygen saturation of 20% or less. In another aspect, the OR and OCR stored blood has an initial oxygen saturation of 10% or less. In another aspect, the OR and OCR stored blood has an initial oxygen saturation of 5% or less. In another aspect, the OR and OCR stored blood has an initial oxygen saturation of 3% or less.

[0034] In one aspect of the present disclosure, the OCR stored blood for use in transfusion therapy of trauma patients in need of transfusion therapy has an initial pCO2 (at 37 °C) between 10 and 40 mmHg. In another aspect, the OCR stored blood has an initial pCO2 between 10 and 30 mmHg. In another aspect, the OCR stored blood has an initial pCO2 between 10 and 20 mmHg. In another aspect, the OCR stored blood has an initial pCO2 between 10 and 15 mmHg. In yet another aspect, the OCR stored blood has an initial pCO2 of less than 10 mmHg.

[0035] In one aspect of the present disclosure, the OR and OCR stored blood for use in transfusion therapy of trauma patients in need of transfusion therapy has an initial oxygen saturation of 20% or less and is stored for less than 2 days. In one aspect, the OR and OCR stored blood has an initial oxygen saturation of 20% or less and is stored for less than 7 days. In another aspect, the OR and OCR stored blood has an initial oxygen saturation of 20% or less and is stored for less than 14 days. In another aspect, the oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 21 days. In another aspect, the oxygen-reduced stored blood for use in transfusion of trauma patients in need of transfusion therapy has an initial oxygen saturation of 20% or less and is stored for less than 28 days. In another aspect, the oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 35 days. In another aspect, the oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 42 days. In another aspect, the oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 45 days. In one aspect of the present disclosure, the OR and OCR stored blood has an oxygen saturation of 20% or less during storage.

[0036] Suitable blood for use in a method for use in transfusion therapy of a trauma patient in need of transfusion therapy in the present disclosure includes oxygen-reduced stored blood having an anticoagulant. In one aspect of the present disclosure, the oxygen-reduced red blood cells are stored for up to 3 weeks to produce the oxygen-reduced stored blood. In another aspect, the oxygen-reduced stored blood typically further includes an additive solution. Suitable additive solutions in the present disclosure include AS-1, AS-3 (Nutricel®), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, OFAS3, and combinations thereof. In one aspect, the additive solution is added during separation of the components. In one aspect, the additive solution is AS-1. In another aspect, the additive solution is AS-3. In another aspect, the additive solution is SAGM.

[0037] The method of the present disclosure is to increase the mean arterial pressure (MAP) in a trauma patient in need of transfusion therapy, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and increasing the mean arterial pressure, including this. In one aspect, the mean arterial pressure increases between 20% and 60%. In another aspect, the mean arterial pressure increases between 30% and 60%. In another aspect, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood increases between 30% and 50%. In yet another aspect, the mean arterial pressure increases between 30% and 60%. In a further aspect, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood increases between 30% and 40%. In one aspect, the mean arterial pressure increases by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90% more than the mean arterial pressure of a patient transfused with conventional stored blood.

[0038] In one aspect of the present disclosure, the mean arterial pressure increases by at least 1.5-fold. In another aspect, the mean arterial pressure of trauma patients receiving OR or OCR blood transfusion therapy increases by at least 2-fold. In a further aspect, the mean arterial pressure increases between 1 and 2-fold. In one aspect of the present disclosure, the mean arterial pressure of trauma patients receiving OR or OCR blood transfusion therapy increases by at least 10 mmHg, at least 20 mmHg, at least 30 mmHg, at least 40 mmHg, at least 50 mmHg, or at least 60 mmHg. In another aspect, the mean arterial pressure of trauma patients receiving OR or OCR blood transfusion therapy increases between 20 and 50 mmHg. In a further aspect, the mean arterial pressure increases between 30 and 50 mmHg.

[0039] The method of the present disclosure is to increase the mean arterial pressure in trauma patients in need of blood transfusion therapy to between 70 and 110 mmHg, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and increasing the mean arterial pressure. In another aspect, the mean arterial pressure of trauma patients receiving OR or OCR blood transfusion therapy increases to at least 70 mmHg. In another aspect, the mean arterial pressure of trauma patients receiving OR or OCR blood transfusion therapy increases to at least 80 mmHg. In yet another aspect, the mean arterial pressure increases to at least 90 mmHg. In a further aspect, the mean arterial pressure increases to at least 100 mmHg. In one aspect of the present disclosure, the mean arterial pressure in a subject in need thereof is maintained between 70 and 110 mmHg for at least 1 hour after transfusion. In another aspect, the mean arterial pressure is maintained between 70 and 110 mmHg for at least 2 hours after transfusion. In yet another aspect, the mean arterial pressure is maintained between 70 and 105 mmHg for at least 3 hours after transfusion. In another aspect, the mean arterial pressure is maintained between 70 and 110 mmHg for at least 4 hours after transfusion. In another aspect, the mean arterial pressure is maintained between 70 and 110 mmHg for at least 5 hours after transfusion.

[0040] The method of the present disclosure provides for increasing the mean arterial pressure in trauma patients in need of blood transfusion therapy at a faster rate than the mean arterial pressure in patients transfused with conventional stored blood, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and includes this. In one aspect, the mean arterial pressure of patients transfused with OR or OCR blood recovers within normal physiological parameters in half the time compared to conventional stored blood.

[0041] The method of the present disclosure provides for reducing the amount of stored blood required for transfusion in trauma patients in need of blood transfusion therapy, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and includes this. In one aspect, the amount of OR stored blood required for transfusion is 10 to 90% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion is 10 to 30% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion is 20 to 50% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion is 20 to 80% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion is 30 to 80% less than the amount of conventional stored blood required. In yet another aspect, the amount of OR stored blood required for transfusion is 40 to 85% less than the amount of conventional stored blood required. In a further aspect, the amount of OR stored blood required for transfusion is 50 to 90% less than the amount of conventional stored blood required.

[0042] The method of the present disclosure provides for reducing, which includes reducing the amount of stored blood required for transfusion in trauma patients who require transfusion therapy by at least 10%, and supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the amount of OR stored blood required for transfusion is at least 20% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion is at least 30% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion is at least 40% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion is at least 50% less than the amount of conventional stored blood required. In yet another aspect, the amount of OR stored blood required for transfusion is at least 60% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion is at least 70% less than the amount of conventional stored blood required. In a further aspect, the amount of OR stored blood required for transfusion is between about 10 - 20%, about 20 - 30%, about 30 - 40%, about 40 - 50%, about 50 - 60%, about 60 - 70%, about 70 - 80%, about 80% - 90%, or about 90 - 95% less than the amount of conventional stored blood required. In another aspect, the amount of OR stored blood required for transfusion therapy in trauma patients who require transfusion therapy is between 10 - 20%, 20 - 30%, 30 - 40%, 40 - 50%, 50 - 60%, 60 - 70%, 70 - 80%, 80% - 90%, or 90 - 95% less than the amount of conventional stored blood required.

[0043] Lactate clearance is a biomarker for resuscitation from hemorrhagic shock. See: Hashmi et al., “Predictors of mortality in geriatric trauma patients: a systematic review and meta-analysis,” The journal of trauma and acute care surgery, 76:894-901 (2014); Regnier et al., “Prognostic significance of blood lactate and lactate clearance in trauma patients,” Anesthesiology, 117:1276-88 (2012); and Zhang et al., “Lactate clearance is a useful biomarker for the prediction of all-cause mortality in critically ill patients: a systematic review and meta-analysis,” Critical care medicine, 42:2118-25 (2014) (the “Zhang 2014”) (each of which is incorporated herein by reference in its entirety). The clinical value of lactate clearance is useful for predicting the outcome of patients with septic shock and patients in critical conditions without obvious circulatory shock. An increase in lactate is an indicator of adverse clinical outcomes, and rapid clearance of lactate is widely associated with improved outcomes in heterogeneous ICU or ED patient populations. See Zhang 2014. The decrease in lactate levels in animals resuscitated using OR-RBC compared to conventional RBC supports the idea that resuscitation with OR RBC may significantly improve the clinical outcome of patients. See FIGS. 7A and 7B.

[0044] The method of the present disclosure is to reduce the lactate level in trauma patients who require blood transfusion therapy, including supplying the trauma patients with oxygen-reduced (OR) stored blood having an oxygen saturation of 20% or less before and during storage, and includes this. In one aspect, the lactate level is reduced between 10% and 90%. In one aspect, transfusion with OR stored blood reduces the lactate level in trauma patients who require blood transfusion therapy between 10% and 50%. In another aspect, transfusion with OR stored blood reduces the lactate level in trauma patients who require blood transfusion therapy between 20% and 40%. In another aspect, transfusion with OR stored blood reduces the lactate level in trauma patients who require blood transfusion therapy between 50% and 90%. In yet another aspect, transfusion with OR stored blood reduces the lactate level in trauma patients who require blood transfusion therapy between 60% and 90%. In another aspect, transfusion with OR stored blood reduces the lactate level in trauma patients who require blood transfusion therapy between 10% and 20%, 20% and 30%, 30% and 40%, 40% and 50%, 50% and 60%, 60% and 70%, 70% and 80%, or 80% and 90%. In another aspect, transfusion with OR stored blood reduces the lactate level in trauma patients who require blood transfusion therapy by at least 10%. In another aspect, transfusion with OR stored blood reduces the lactate level in trauma patients who require blood transfusion therapy by at least 20%. In a further aspect, transfusion with OR stored blood reduces the lactate level in trauma patients who require blood transfusion therapy by at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, or at least 90%.

[0045] The method of the present disclosure is to reduce elevated lactate levels in trauma patients in need of blood transfusion therapy to between about 0.5 and about 2.5 mmol / L, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and reducing, including this. In one aspect, the lactate level in a trauma patient in need of blood transfusion therapy is reduced to between about 0.9 and about 2 mmol / L. In one aspect, the lactate level in a trauma patient in need of blood transfusion therapy is reduced to between about 0.9 and about 1.7 mmol / L. In another aspect, the lactate level in a patient in need of blood transfusion therapy is reduced to between about 1.4 and about 2.4 mmol / L. In another aspect, the lactate level in a trauma patient in need of blood transfusion therapy is reduced to between about 1.7 and about 2.5 mmol / L. In yet another aspect, the lactate level in a trauma patient in need of blood transfusion therapy is reduced to less than about 2.5 mmol / L. In a further aspect, the lactate level in a trauma patient in need of blood transfusion therapy is reduced to less than about 2.0 mmol / L. In another aspect, the lactate level in a trauma patient in need of blood transfusion therapy is reduced to less than about 1.5 mmol / L. In another aspect, the lactate level in a trauma patient in need of blood transfusion therapy is reduced to less than about 1.0 mmol / L. In yet another aspect, the lactate level in a trauma patient in need of blood transfusion therapy is reduced to between about 0.5 and about 1.0 mmol / L.

[0046] The method of the present disclosure is to reduce elevated lactate levels in trauma patients requiring blood transfusion therapy to between 0.5 and 2.5 mmol / L, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and reducing, including this. In one aspect, the lactate level in a patient requiring blood transfusion therapy is reduced to between 0.9 and 2 mmol / L. In one aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to between 0.9 and 1.7 mmol / L. In another aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to between 1.4 and 2.4 mmol / L. In another aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to between 1.7 and 2.5 mmol / L. In another aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to between 0.5 and 1 mmol / L.

[0047] The method of the present disclosure is to reduce elevated lactate levels in hemorrhagic trauma patients requiring blood transfusion therapy to less than 4 mmol / L, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and reducing, including this. In one aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to less than 3 mmol / L. In yet another aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to less than 2.5 mmol / L. In another aspect, the lactate level in a patient is reduced to less than 2.3 mmol / L. In another aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to less than 2 mmol / L. In another aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to less than 2 mmol / L. In another aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to less than 1.5 mmol / L. In another aspect, the lactate level in a trauma patient requiring blood transfusion therapy is reduced to less than 1 mmol / L.

[0048] Blood glucose levels are also known to be predictors of outcomes in several disease patterns, particularly in trauma patients. Trauma patients are more likely to have poor outcomes than patients with other critical conditions due to hyperglycemia. See Kreutziger et al., “Admission blood glucose predicted hemorrhagic shock in multiple trauma patients,” Injury, 46:15-20 (2015) (which is hereby incorporated by reference in its entirety). In studies evaluating the relationship between early hyperglycemia and trauma patients, early hyperglycemia was verified at three possible cutoffs: glucose >110 mg / dL, glucose >150 mg / dL, and glucose >200 mg / dL. See Laird et al., “Relationship of early hyperglycemia to mortality in trauma patients,” J Trauma, 56:1058-62 (2004) (which are hereby incorporated by reference in their entirety). Glucose levels >200 mg / dL are associated with significantly higher infection rates and mortality in trauma patients, regardless of injury characteristics, which was not the case at the 110 mg / dL or 150 mg / dL cutoffs. The decrease in glucose levels in animals resuscitated using OR and OR-RBC compared to those using conventional RBC supports the idea that resuscitation with OR-RBC may significantly improve the clinical outcomes of patients. See FIGS. 8A and 8B.

[0049] The method of the present disclosure is to reduce glucose in trauma patients who require blood transfusion therapy, including supplying oxygen-reduced (OR) stored blood having an oxygen saturation of 20% or less before and during storage to the trauma patients, and includes reducing. In one aspect, the glucose is reduced between 10% and 90% compared to transfusion of blood stored under conventional conditions. In one aspect, transfusion with OR stored blood reduces glucose between 10% and 50% compared to transfusion of blood stored under conventional conditions. In another aspect, transfusion with OR stored blood reduces glucose between 20% and 40% compared to transfusion of blood stored under conventional conditions. In another aspect, transfusion with OR stored blood reduces glucose between 50% and 90% compared to transfusion of blood stored under conventional conditions. In yet another aspect, transfusion with OR stored blood reduces glucose between 60% and 90%. In another aspect, transfusion with OR stored blood reduces glucose between 10% and 20%, 20% and 30%, 30% and 40%, 40% and 50%, 50% and 60%, 60% and 70%, 70% and 80%, or 80% and 90% compared to transfusion of blood stored under conventional conditions. In another aspect, transfusion with OR stored blood reduces glucose by at least 10% compared to transfusion of blood stored under conventional conditions. In another aspect, transfusion with OR stored blood reduces glucose by at least 20%. In a further aspect, transfusion with OR stored blood reduces glucose by at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, or at least 90%.

[0050] The method of the present disclosure is to reduce the glucose level in trauma patients in need of blood transfusion therapy to between about 70 and about 120 mg / dL, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and to provide and include reducing. In one aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is between about 70 and about 110 mg / dL. In another aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is between about 70 and about 100 mg / dL. In another aspect, the glucose in a trauma patient after blood transfusion therapy using OR or OCR blood is between about 90 and about 120 mg / dL. In another aspect, the glucose in a trauma patient after blood transfusion therapy using OR or OCR blood is between about 90 and about 100 mg / dL.

[0051] The method of the present disclosure is to reduce the glucose level in trauma patients in need of blood transfusion therapy to between 70 and 120 mg / dL, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and to provide and include reducing. In one aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is between 70 and 110 mg / dL. In another aspect, the glucose in the patient is between 70 and 100 mg / dL. In another aspect, the glucose in the patient is between 90 and 120 mg / dL. In another aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is between 90 and 100 mg / dL.

[0052] The method of the present disclosure provides for reducing the glucose level in a trauma patient in need of blood transfusion therapy to less than 120 mg / dL, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In a further aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is less than 110 mmol / L. In another aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is less than 100 mg / dL. In another aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is less than 200 mg / dL. In another aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is less than 90 mg / dL. In another aspect, the glucose in a patient after blood transfusion therapy using OR or OCR blood is less than 80 mg / dL.

[0053] In one aspect of the present disclosure, trauma patients have an increased risk of complications from transfusion therapy based on an existing or baseline condition. In one aspect, a trauma patient has an existing or baseline condition selected from the group consisting of diabetes, ischemic heart disease, systemic inflammatory response syndrome caused by trauma or infection, multiple organ failure caused by trauma or infection, smoking, chronic obstructive pulmonary disease, such as systemic inflammation due to infection, coagulopathy, and autoimmune disease. In another aspect, a trauma patient has one or more existing or baseline conditions selected from the group consisting of diabetes, ischemic heart disease, systemic inflammatory response syndrome caused by trauma or infection, multiple organ failure caused by trauma or infection, smoking, and chronic obstructive pulmonary disease, such as systemic inflammation due to infection, coagulopathy, and autoimmune disease. In another aspect, a trauma patient has two or more existing or baseline conditions selected from the group consisting of diabetes, ischemic heart disease, systemic inflammatory response syndrome caused by trauma or infection, multiple organ failure caused by trauma or infection, smoking, chronic obstructive pulmonary disease, such as systemic inflammation due to infection, coagulopathy, and autoimmune disease. In another aspect, a trauma patient has three or more existing or baseline conditions selected from the group consisting of diabetes, ischemic heart disease, systemic inflammatory response syndrome caused by trauma or infection, multiple organ failure caused by trauma or infection, smoking, chronic obstructive pulmonary disease, such as systemic inflammation due to infection, coagulopathy, and autoimmune disease.

[0054] Patients experience adverse events, including liver injury or liver failure, kidney injury or kidney failure, lung injury or lung failure, or combinations thereof, during hemorrhagic shock. The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits one or more adverse events selected from the group consisting of liver injury or liver failure, kidney injury or kidney failure, or lung injury or lung failure, and includes this. The present disclosure provides that a patient in need of transfusion therapy using OR or OCR RBCs exhibits two or more adverse events selected from the group consisting of liver injury or liver failure, kidney injury or kidney failure, or lung injury or lung failure, and includes this.

[0055] The method of the present disclosure is to reduce adverse events in trauma patients, including supplying trauma patients in need of blood transfusion therapy with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and includes reducing. In one aspect, the adverse events after transfusion using OR or OCR blood are reduced by at least 5%. In another aspect, the adverse events after transfusion using OR or OCR blood are reduced by at least 10%. In another aspect, the adverse events after transfusion using OR or OCR blood are reduced by at least 20%. In another aspect, the adverse events after transfusion using OR or OCR blood are reduced by at least 30%. In another aspect, the adverse events after transfusion using OR or OCR blood are reduced by at least 40%. In another aspect, the adverse events after transfusion using OR or OCR blood are reduced by at least 50%. In another aspect, the adverse events after transfusion using OR or OCR blood are reduced by at least 60%. In another aspect, the adverse events are reduced by at least 70%. In another aspect, the adverse events after transfusion using OR or OCR blood are reduced by at least 80%. In another aspect, the adverse events are reduced by at least 90%. In a further aspect, the adverse events after blood transfusion therapy using OR or OCR blood are reduced between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95%. In one aspect, the adverse events after transfusion using OR or OCR blood are liver injury or liver damage. In another aspect, the adverse events are lung injury or lung damage. In yet another aspect, the adverse events are kidney injury or kidney damage. In a further aspect, the adverse events are liver injury, lung injury, kidney injury, or a combination thereof.

[0056] An increase in the levels of liver enzymes (including, but not limited to, aspartate aminotransferase (AST) and alanine aminotransferase (ALT)) means some form of liver damage, shock, or injury. The method of the present disclosure provides for reducing elevated levels of liver enzymes in trauma patients, including by supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage.

[0057] The method of the present disclosure provides for reducing the AST level in trauma patients in need of blood transfusion therapy, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the AST level is reduced by at least 5% compared to the AST level of patients transfused with conventional stored blood. In another aspect, the AST level is reduced by at least 10% compared to the AST level of patients transfused with conventional stored blood. In another aspect, the AST level is reduced by at least 20% compared to the AST level of patients transfused with conventional stored blood. In another aspect, the AST level is reduced by at least 30% compared to the AST level of patients transfused with conventional stored blood. In another aspect, the AST level is reduced by at least 40%. In another aspect, the AST level is reduced by at least 50% compared to the AST level of patients transfused with conventional stored blood. In another aspect, the AST level is reduced by at least 60%. In another aspect, the AST level is reduced by at least 70% compared to the AST level of patients transfused with conventional stored blood. In yet another aspect, the AST level is reduced by at least 80%. In a further aspect, the AST level is reduced by at least 90% compared to the AST level of patients transfused with conventional stored blood. In a further aspect, the AST level is reduced between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the AST level of patients transfused with conventional stored blood.

[0058] The method of the present disclosure is to reduce the AST level in trauma patients who require blood transfusion therapy between 1.5 to 10 times, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and providing and including this reduction. In one aspect, the AST level is reduced between 2 to 3 times in comparison with the AST level of patients transfused with conventional stored blood. In another aspect, the AST level is reduced between 3 to 4 times. In another aspect, the AST level is reduced between 4 to 10 times. In another aspect, the AST level is reduced between 6 to 9 times in comparison with the AST level of patients transfused with conventional stored blood. In a further aspect, the AST level is reduced between 2 to 5 times. In another aspect, the AST level is reduced between 10 to 50 times in comparison with the AST level of patients transfused with conventional stored blood.

[0059] The method of the present disclosure provides for reducing, in a trauma patient in need of blood transfusion therapy, the AST level by at least 1.5-fold, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the AST level is reduced by at least 2-fold as compared to the AST level of a patient transfused with conventionally stored blood. In another aspect, the AST level is reduced by at least 3-fold as compared to the AST level of a patient transfused with conventionally stored blood. In another aspect, the AST level is reduced by at least 4-fold as compared to the AST level of a patient transfused with conventionally stored blood. In another aspect, the AST level is reduced by at least 5-fold as compared to the AST level of a patient transfused with conventionally stored blood. In a further aspect, the AST level is reduced by at least 6-fold. In another aspect, the AST level is reduced by at least 7-fold as compared to the AST level of a patient transfused with conventionally stored blood. In another aspect, the AST level is reduced by at least 8-fold. In another aspect, the AST level is reduced by at least 9-fold as compared to the AST level of a patient transfused with conventionally stored blood. In another aspect, the AST level is reduced by at least 10-fold as compared to the AST level of a patient transfused with conventionally stored blood. In a further aspect, the AST level is reduced by at least 50-fold as compared to the AST level of a patient transfused with conventionally stored blood.

[0060] The method of the present disclosure is to reduce the ALT level in trauma patients who require blood transfusion therapy, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and includes reducing. In one aspect, the ALT level is reduced by at least 5% compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced by at least 10% compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced by at least 20% compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced by at least 30% compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced by at least 40% compared to the AS ALT T level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced by at least 50% compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced by at least 60% compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced by at least 70% compared to the ALT level of patients transfused with conventional stored blood. In yet another aspect, the ALT level is reduced by at least 80% compared to the ALT level of patients transfused with conventional stored blood. In a further aspect, the ALT level is reduced by at least 90%. In a further aspect, the ALT level is reduced between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the ALT level of patients transfused with conventional stored blood.

[0061] The method of the present disclosure is to reduce the ALT level in trauma patients in need of blood transfusion therapy between 1.5 and 10 times, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and providing for the reduction, including this. In one aspect, the ALT level is reduced between 2 and 3 times compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced between 3 and 4 times compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced between 4 and 10 times compared to the ALT level of patients transfused with conventional stored blood. In another aspect, the ALT level is reduced between 6 and 9 times compared to the ALT level of patients transfused with conventional stored blood. In a further aspect, the ALT level is reduced between 2 and 5 times. In another aspect, the ALT level is reduced between 10 and 50 times compared to the ALT level of patients transfused with conventional stored blood.

[0062] The method of the present disclosure provides for reducing, in a trauma patient in need of blood transfusion therapy, the ALT level by at least 1.5-fold, including supplying to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the ALT level is reduced by at least 2-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In another aspect, the ALT level is reduced by at least 3-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In another aspect, the ALT level is reduced by at least 4-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In another aspect, the ALT level is reduced by at least 5-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In a further aspect, the ALT level is reduced by at least 6-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In another aspect, the ALT level is reduced by at least 7-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In another aspect, the ALT level is reduced by at least 8-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In another aspect, the ALT level is reduced by at least 9-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In another aspect, the ALT level is reduced by at least 10-fold as compared to the ALT level of a patient transfused with conventionally stored blood. In a further aspect, the ALT level is reduced by at least 50-fold as compared to the ALT level of a patient transfused with conventionally stored blood.

[0063] Markers of kidney function during and after hemorrhagic trauma include urinary neutrophil gelatinase-associated lipocalin (u-NGAL), serum creatinine, and blood urea nitrogen (BUN). See Treeprasertsuk et al., “Urine neutrophil gelatinase-associated lipocalin: a diagnostic and prognostic marker for acute kidney injury (AKI) in hospitalized cirrhotic patients with AKI-prone conditions,” BMC Gastroenterol 15:140 (2015) (which is hereby incorporated by reference in its entirety). Gene expression analysis reported in over 150 different studies conducted in AKI models from several species from rodents to humans has consistently revealed that the NGAL gene is one of the most dramatically upregulated genes in the kidney immediately after the onset of ischemia or nephrotoxicity. See Ciccia et al., “Pediatric acute kidney injury: prevalence, impact and management challenges,” Int J Nephrol Renovasc Dis, 10:77-84 (2017) (which is hereby incorporated by reference in its entirety). Similarly, serum creatinine levels can vary depending on age, race, and body size, but elevated creatinine levels indicate kidney injury. Creatinine levels above 1.2 in women and above 1.4 in men may be early signs of kidney injury. An increase in blood urea nitrogen (BUN) is seen with kidney disease or renal failure, as well as with congestive heart failure, shock, and bleeding within the gastrointestinal tract. A BUN level higher than 100 mg / dL indicates severe kidney injury. A decrease in BUN level is also a concern, as it may indicate fluid overload, trauma, surgery, opioids, malnutrition, and the use of anabolic steroids.See Pagana, “Mosby’s Manual of Diagnostic and Laboratory Tests,” St. Louis Mosby, Inc., (1998); and Gowda, et al., “Markers of renal function tests,” NAm JMed Sci. 2(4):170-173 (2010) (the entireties of which are incorporated herein by reference). The u-NGAL decrease (Figures 16A and 16B), serum creatinine (Figures 11A and 11B), and BUN (Figures 12A and 12B) levels in animals resuscitated with OR- and OCR-RBC compared to conventional RBCs, provided by the present disclosure, indicate that resuscitation with OR-RBC can significantly improve the clinical outcome of patients.

[0064] The method of the present disclosure is to reduce the urinary neutrophil gelatinase-associated lipocalin (u-NGAL) level in trauma patients who require blood transfusion therapy, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and reducing the u-NGAL level. In one aspect, the u-NGAL level is reduced by at least 5% compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 10% compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 20% compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 30%. In another aspect, the u-NGAL level is reduced by at least 40% compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 50% compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 60% compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 70% compared to the u-NGAL level of patients transfused with conventional stored blood. In yet another aspect, the u-NGAL level is reduced by at least 80% compared to the u-NGAL level of patients transfused with conventional stored blood. In a further aspect, the u-NGAL level is reduced by at least 90% compared to the u-NGAL level of patients transfused with conventional stored blood. In a further aspect, the u-NGAL level is reduced between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the u-NGAL level of patients transfused with conventional stored blood.

[0065] The method of the present disclosure is to reduce the urinary neutrophil gelatinase-associated lipocalin (u-NGAL) level in trauma patients who require blood transfusion therapy, including supplying oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to the trauma patients at 1.5 to 10 times, and providing a reduction, including this. In one aspect, the u-NGAL level is reduced between 2 and 3 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced between 3 and 4 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced between 4 and 10 times. In another aspect, the u-NGAL level is reduced between 6 and 9 times compared to the u-NGAL level of patients transfused with conventional stored blood. In a further aspect, the u-NGAL level is reduced between 2 and 5 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced between 10 and 50 times compared to the u-NGAL level of patients transfused with conventional stored blood.

[0066] The method of the present disclosure is to reduce the urinary neutrophil gelatinase-associated lipocalin (u-NGAL) level in trauma patients who require blood transfusion therapy, including supplying oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to the trauma patients at least 1.5 times, and providing and including reducing. In one aspect, the u-NGAL level is reduced by at least 2 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 3 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 4 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 5 times compared to the u-NGAL level of patients transfused with conventional stored blood. In a further aspect, the u-NGAL level is reduced by at least 6 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 7 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 8 times compared to the u-NGAL level of patients transfused with conventional stored blood. In another aspect, the u-NGAL level is reduced by at least 9 times. In another aspect, the u-NGAL level is reduced by at least 10 times. In a further aspect, the u-NGAL level is reduced by at least 50 times.

[0067] The method of the present disclosure is to reduce serum creatinine levels in trauma patients who require blood transfusion therapy, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and includes reducing. In one aspect, the serum creatinine level is reduced by at least 5% compared to the serum creatinine level of patients transfused with conventional stored blood. In another aspect, the serum creatinine level is reduced by at least 10% compared to the serum creatinine level of patients transfused with conventional stored blood. In another aspect, the serum creatinine level is reduced by at least 20% compared to the serum creatinine level of patients transfused with conventional stored blood. In another aspect, the serum creatinine level is reduced by at least 30% compared to the serum creatinine level of patients transfused with conventional stored blood. In another aspect, the serum creatinine level is reduced by at least 40% compared to the serum creatinine level of patients transfused with conventional stored blood. In another aspect, the serum creatinine level is reduced by at least 50% compared to the serum creatinine level of patients transfused with conventional stored blood. In another aspect, the serum creatinine level is reduced by at least 60% compared to the serum creatinine level of patients transfused with conventional stored blood. In another aspect, the serum creatinine level is reduced by at least 70% compared to the serum creatinine level of patients transfused with conventional stored blood. In yet another aspect, the serum creatinine level is reduced by at least 80% compared to the serum creatinine level of patients transfused with conventional stored blood. In a further aspect, the serum creatinine level is reduced by at least 90% compared to the serum creatinine level of patients transfused with conventional stored blood. In a further aspect, the serum creatinine level is reduced between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the serum creatinine level of patients transfused with conventional stored blood.

[0068] The method of the present disclosure is to reduce serum creatinine levels in trauma patients who require blood transfusion therapy between 1.5 and 10 times, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and reducing the levels. In one aspect, the serum creatinine level is reduced between 2 and 3 times compared to the serum creatinine level of patients transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced between 3 and 4 times compared to the serum creatinine level of patients transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced between 4 and 10 times compared to the serum creatinine level of patients transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced between 6 and 9 times compared to the serum creatinine level of patients transfused with conventionally stored blood. In a further aspect, the serum creatinine level is reduced between 2 and 5 times compared to the serum creatinine level of patients transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced between 10 and 50 times compared to the serum creatinine level of patients transfused with conventionally stored blood.

[0069] The method of the present disclosure provides for reducing, in a trauma patient in need of blood transfusion therapy, serum creatinine levels by at least 1.5-fold, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the serum creatinine level is reduced by at least 2-fold as compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 3-fold as compared to the AST level of a patient transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 4-fold as compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 5-fold. In a further aspect, the serum creatinine level is reduced by at least 6-fold as compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 7-fold as compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 8-fold as compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 9-fold as compared to the serum creatinine level of a patient transfused with conventionally stored blood. In another aspect, the serum creatinine level is reduced by at least 10-fold as compared to the serum creatinine level of a patient transfused with conventionally stored blood.

[0070] The method of the present disclosure is to reduce the serum creatinine level in trauma patients who require blood transfusion therapy to between 0.5 and 1.5 mg / dL, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and reducing the serum creatinine level. In one aspect, the serum creatinine level is reduced to between 0.5 and 1 mg / dL in comparison with the serum creatinine level of patients transfused with conventional stored blood. In one aspect, the serum creatinine level is reduced to between 0.8 and 1 mg / dL in comparison with the serum creatinine level of patients transfused with conventional stored blood. In another aspect, the serum creatinine level is reduced to between 0.7 and 1.5 mg / dL in comparison with the serum creatinine level of patients transfused with conventional stored blood.

[0071] The method of the present disclosure is to reduce the serum creatinine level in trauma patients who require blood transfusion therapy to less than 1.5 mg / dL, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and reducing the serum creatinine level. In one aspect, the serum creatinine level is reduced to less than 1.4 mg / dL in comparison with the serum creatinine level of patients transfused with conventional stored blood. In one aspect, the serum creatinine level is reduced to less than 1 mg / dL. In another aspect, the serum creatinine level is reduced to less than 0.8 mg / dL in comparison with the serum creatinine level of patients transfused with conventional stored blood.

[0072] The method of the present disclosure is to reduce the BUN level in trauma patients, including supplying oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to trauma patients who require blood transfusion therapy, and to provide a reduction. In one aspect, the BUN level is reduced by at least 5% compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced by at least 10% compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced by at least 20% compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced by at least 30% compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced by at least 40% compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced by at least 50% compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced by at least 60% compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced by at least 70% compared to the BUN level of patients transfused with conventional stored blood. In yet another aspect, the BUN level is reduced by at least 80% compared to the BUN level of patients transfused with conventional stored blood. In a further aspect, the BUN level is reduced by at least 90% compared to the BUN level of patients transfused with conventional stored blood. In a further aspect, the BUN level is reduced between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the BUN level of patients transfused with conventional stored blood.

[0073] The method of the present disclosure provides for reducing the BUN level in trauma patients by between 1.5 and 10 times, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the BUN level is reduced between 2 and 3 times as compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced between 3 and 4 times. In another aspect, the BUN level is reduced between 4 and 10 times as compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced between 6 and 9 times as compared to the BUN level of patients transfused with conventional stored blood. In a further aspect, the BUN level is reduced between 2 and 5 times as compared to the BUN level of patients transfused with conventional stored blood. In another aspect, the BUN level is reduced between 10 and 100 times as compared to the BUN level of patients transfused with conventional stored blood.

[0074] The method of the present disclosure provides for reducing, which includes reducing the BUN level in trauma patients by at least 1.5-fold, and supplying trauma patients in need of blood transfusion therapy with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the BUN level is reduced by at least 2-fold as compared to the BUN level of patients transfused with conventionally stored blood. In another aspect, the BUN level is reduced by at least 3-fold as compared to the BUN level of patients transfused with conventionally stored blood. In another aspect, the BUN level is reduced by at least 4-fold as compared to the BUN level of patients transfused with conventionally stored blood. In another aspect, the BUN level is reduced by at least 5-fold as compared to the BUN level of patients transfused with conventionally stored blood. In a further aspect, the BUN level is reduced by at least 6-fold. In another aspect, the BUN level is reduced by at least 7-fold as compared to the BUN level of patients transfused with conventionally stored blood. In another aspect, the BUN level is reduced by at least 8-fold as compared to the BUN level of patients transfused with conventionally stored blood. In another aspect, the BUN level is reduced by at least 9-fold as compared to the BUN level of patients transfused with conventionally stored blood. In another aspect, the BUN level is reduced by at least 10-fold as compared to the BUN level of patients transfused with conventionally stored blood.

[0075] The method of the present disclosure provides for reducing the percentage of CD45+ neutrophils in trauma patients, which includes supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the percentage of CD45+ neutrophils is reduced by at least 5% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 10% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 20% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 30% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 40% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 50% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 60% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 70% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In yet another aspect, the percentage of CD45+ neutrophils is reduced by at least 80% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In a further aspect, the percentage of CD45+ neutrophils is reduced by at least 90% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood.In a further aspect, the percentage of CD45+ neutrophils is reduced between 1 - 10%, 10 - 20%, 20 - 30%, 30 - 40%, 40 - 50%, 50 - 60%, 60 - 70%, 70 - 80%, 80 - 90%, or 90 - 95% compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood.

[0076] The method of the present disclosure provides for reducing the percentage of CD45+ neutrophils in trauma patients in need of blood transfusion therapy between 1.5 - 10 fold, including supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the percentage of CD45+ neutrophils is reduced between 2 - 3 fold compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced between 3 - 4 fold compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced between 4 - 10 fold compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced between 6 - 9 fold compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In a further aspect, the percentage of CD45+ neutrophils is reduced between 2 - 5 fold compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced between 10 - 50 fold compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood.

[0077] The method of the present disclosure provides for reducing, in a trauma patient in need of blood transfusion therapy, the percentage of CD45+ neutrophils by at least 1.5-fold, including supplying to the trauma patient oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the percentage of CD45+ neutrophils is reduced by at least 2-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 3-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 4-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 5-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In a further aspect, the percentage of CD45+ neutrophils is reduced by at least 6-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 7-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 8-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 9-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood. In another aspect, the percentage of CD45+ neutrophils is reduced by at least 10-fold as compared to the CD45+ neutrophil levels in patients transfused with conventional stored blood.

[0078] The method of the present disclosure is to reduce CXCL1 levels in trauma patients in need of blood transfusion therapy, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and includes reducing. In one aspect, the CXCL1 level is reduced by at least 5% compared to the CXCL1 level of patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced by at least 10% compared to the CXCL1 level of patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced by at least 20% compared to the CXCL1 level of patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced by at least 30% compared to the CXCL1 level of patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced by at least 40% compared to the CXCL1 level of patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced by at least 50% compared to the CXCL1 level of patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced by at least 60% compared to the CXCL1 level of patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced by at least 70% compared to the CXCL1 level of patients transfused with conventional stored blood. In yet another aspect, the CXCL1 level is reduced by at least 80% compared to the CXCL1 level of patients transfused with conventional stored blood. In a further aspect, the CXCL1 level is reduced by at least 90% compared to the CXCL1 level of patients transfused with conventional stored blood. In a further aspect, the CXCL1 level is reduced between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the CXCL1 level of patients transfused with conventional stored blood.

[0079] The method of the present disclosure is to reduce CXCL1 levels in trauma patients between 1.5 and 10-fold, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, to provide for reducing, and including this. In one aspect, the CXCL1 level is reduced between 2 and 3-fold compared to the CXCL1 level in patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced between 3 and 4-fold compared to the CXCL1 level in patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced between 4 and 10-fold compared to the CXCL1 level in patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced between 6 and 9-fold compared to the CXCL1 level in patients transfused with conventional stored blood. In a further aspect, the CXCL1 level is reduced between 2 and 5-fold compared to the CXCL1 level in patients transfused with conventional stored blood. In another aspect, the CXCL1 level is reduced between 10 and 100-fold compared to the CXCL1 level in patients transfused with conventional stored blood.

[0080] The method of the present disclosure provides for reducing CXCL1 levels in trauma patients by at least 1.5-fold, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the CXCL1 level is reduced by at least 2-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 3-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 4-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 5-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood. In a further aspect, the CXCL1 level is reduced by at least 6-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 7-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 8-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 9-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood. In another aspect, the CXCL1 level is reduced by at least 10-fold as compared to the CXCL1 level in patients transfused with conventionally stored blood.

[0081] The method of the present disclosure is to reduce IL-6 levels in trauma patients who require blood transfusion therapy, including supplying the trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, and includes reducing. In one aspect, the IL-6 level is reduced by at least 5% compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced by at least 10% compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced by at least 20%. In another aspect, the IL-6 level is reduced by at least 30% compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced by at least 40% compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced by at least 50% compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced by at least 60% compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced by at least 70% compared to the IL-6 level of patients transfused with conventional stored blood. In yet another aspect, the IL-6 level is reduced by at least 80% compared to the IL-6 level of patients transfused with conventional stored blood. In a further aspect, the IL-6 level is reduced by at least 90% compared to the IL-6 level of patients transfused with conventional stored blood. In a further aspect, the IL-6 level is reduced between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the IL-6 level of patients transfused with conventional stored blood.

[0082] The method of the present disclosure is to reduce the IL-6 level in trauma patients between 1.5 and 10 times, including supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one aspect, the IL-6 level is reduced between 2 and 3 times compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced between 3 and 4 times compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced between 4 and 10 times. In another aspect, the IL-6 level is reduced between 6 and 9 times compared to the IL-6 level of patients transfused with conventional stored blood. In a further aspect, the IL-6 level is reduced between 2 and 5 times compared to the IL-6 level of patients transfused with conventional stored blood. In another aspect, the IL-6 level is reduced between 10 and 100 times compared to the IL-6 level of patients transfused with conventional stored blood.

[0083] The method of the present disclosure provides for reducing, which includes reducing the IL-6 level in trauma patients by at least 1.5-fold, by supplying oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to trauma patients who require blood transfusion therapy. In one aspect, the IL-6 level is reduced by at least 2-fold as compared to the IL-6 level of patients transfused with conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 3-fold as compared to the IL-6 level of patients transfused with conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 4-fold as compared to the IL-6 level of patients transfused with conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 5-fold. In a further aspect, the IL-6 level is reduced by at least 6-fold as compared to the IL-6 level of patients transfused with conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 7-fold as compared to the IL-6 level of patients transfused with conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 8-fold as compared to the IL-6 level of patients transfused with conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 9-fold as compared to the IL-6 level of patients transfused with conventionally stored blood. In another aspect, the IL-6 level is reduced by at least 10-fold as compared to the IL-6 level of patients transfused with conventionally stored blood.

[0084] As used herein, the terms "higher", "greater", or "increased" mean that the measured value of blood that has undergone oxygen reduction and anaerobic storage is at least 1 standard deviation greater for each measurement condition being compared, with a sample size of two or more, when compared to the measured value of conventionally stored blood that has been equivalently processed by another method.

[0085] As used herein, the terms "reduce", "reduced", "lower", "lowered", or "less" mean that a measured value of blood that has undergone oxygen reduction and anaerobic storage is at least 1 standard deviation lower for each measurement condition being compared, with a sample size of five or more, when compared to the measured values of conventional normoxic or hyperoxic stored blood RBCs that have been equivalently processed by another method.

[0086] As used herein, the term "about" refers to ±10%.

[0087] As used herein, "less than" refers to a smaller amount and an amount greater than zero.

[0088] The terms "comprises", "comprising", "includes", "including", "having" and their conjugations mean including, but not limited to, what follows.

[0089] The term "consisting of" means including and limiting what follows.

[0090] The term "consisting essentially of" means that a composition, method, or structure may include additional ingredients, steps, and / or parts, provided that the additional ingredients, steps, and / or parts do not substantially modify the basic and novel characteristics of the claimed composition, method, or structure.

[0091] As used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. For example, "compound" or "at least one compound" can include a plurality of compounds and mixtures thereof.

[0092] As used herein, the term "blood" refers to whole blood, leukoreduced RBC, platelet-reduced RBC, and leukocyte- and platelet-reduced RBC. The term blood further includes concentrated red blood cells, platelet-reduced concentrated red blood cells, leukoreduced concentrated red blood cells, and leukocyte- and platelet-reduced concentrated red blood cells. The temperature of blood can vary depending on the stage of the collection process, starting at 37°C, normal body temperature at the time of collection, but rapidly decreasing to about 30°C when it exits the patient's body, and then further decreasing to room temperature in about 6 hours if no further processing is done, and ultimately being refrigerated between about 4°C and 6°C. In vivo human red blood cells are in a dynamic state. Red blood cells contain hemoglobin, an iron-containing protein that transports oxygen throughout the body and gives red blood its color. The percentage of the blood volume composed of red blood cells is called the hematocrit. As used herein, unless otherwise limited, RBC also includes packed red blood cells (pRBC). Packed red blood cells are prepared from whole blood using centrifugation techniques commonly known in the art. As used herein, unless otherwise indicated, the hematocrit of pRBC is about 70%. As used herein, oxygen-reduced stored RBC may include oxygen- and carbon dioxide-reduced stored RBC. As used herein, oxygen-reduced (OR) blood may include oxygen- and carbon dioxide-reduced (OCR) blood.

[0093] As used herein, the terms "patient" and "subject" are used interchangeably and mean a human or animal in need of a transfusion.

[0094] As used herein, the term "trauma" includes blood loss, hemorrhagic trauma.

[0095] As used herein, the term "hemorrhagic shock" is a shock caused by the loss of circulating blood volume and / or oxygen-carrying capacity. Hemorrhagic shock is caused by any condition associated with blood loss, internal bleeding (e.g., gastrointestinal bleeding) or external bleeding, and in particular trauma (e.g., penetrating trauma or blunt trauma).

[0096] As used herein, the term "adverse event" includes events due to hemorrhagic shock in patients with hemorrhagic trauma.

[0097] As used herein, the terms "injury", "damage", and "failure" refer to organs that are not functioning properly or not functioning as expected in a human or animal without disease or injury.

[0098] As used herein, a "unit" of blood is approximately 450 - 500 ml including anticoagulant. Suitable anticoagulants include CPD, CPDA1, ACD, and ACD - A.

[0099] Throughout this application, various aspects of the present disclosure may be presented in range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the present disclosure. Thus, a description of a range should be considered to specifically disclose all the sub - ranges that could be contemplated, as well as the individual numerical values within that range. For example, a range description such as "from 1 to 6" should be considered to specifically disclose sub - ranges such as "from 1 to 3", "from 1 to 4", "from 1 to 5", "from 2 to 4", "from 2 to 6", "from 3 to 6", as well as the individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0100] When a numerical range is indicated herein, it always means including any number (whether fraction or integer) recited within the indicated range. The expressions "ranging between a first indicated number and a second indicated number" and "ranging from a first indicated number to a second indicated number" are used interchangeably herein and are intended to include the first and second indicated numbers, as well as all fractions and integers therebetween.

[0101] As used herein, the term "method" refers to a manner, means, technique, and procedure for accomplishing a given task, including, but not limited to, supplying a human patient in need of a transfusion with oxygen-depleted stored blood having an initial oxygen saturation of 20% or less and stored for at least two days.

[0102] Although the disclosure has been described with reference to specific embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be used in place of elements of the disclosure without departing from the scope of the disclosure. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the disclosure without departing from its scope.

[0103] Therefore, the disclosure is not intended to be limited to the specific embodiments disclosed as the best mode contemplated for carrying out the disclosure, but rather the disclosure is intended to cover all embodiments falling within the spirit and scope of the appended claims.

Examples

[0104] Example 1: Blood collection and sample preparation Red blood cells from each pool are collected from a total of 12 - 14 rats into CP2D anticoagulant. The pooled blood is leukoreduced using a neonatal leukocyte reduction filter, the components are separated, and the RBCs are stored in AS-3 additive solution. A total of two pools of RBCs are collected. Each pool is divided into four groups: untreated control (C), sham control (SC), oxygen reduction (OR), and oxygen and carbon dioxide reduction (OCR). For the C, SC, OR, and OCR units, the RBC sub-units are processed by transferring them into 80 mL PVC transfusion bags, and the final RBC products are prepared by gas exchange treatment. The RBC bags other than C are filled with 100% N2 (for OR), or 95% N2 / 5% CO2 (for OCR), or air (for SC) through a sterile filter and gently rotated at 2 - 3 RPM on their long sides (for those other than C). For the OR and OCR units, after 10 minutes, the gas is removed through the filter and fresh gas is introduced for subsequent gas exchange treatment. This process is repeated 5 - 8 times until the target SO2% of 5 - 10% is achieved as measured by an ABL-90 CO oximeter (Radiometer Copenhagen). The SC units are rotated without any gas exchange for 60 minutes. The OR and OCR units are stored anaerobically in a canister filled with N2, while the C and SC units are stored in ambient air. All units are stored at 4°C for 3 weeks and samples are taken on days 0, 1, 7, 14, 21, and 28. Two pools are prepared and stored.

[0105] On days 0, 1, 7, 14, 21, and 28, analysis of ATP, 2,3-DPG, and hemolysis is performed. As shown in Figure 1, the ATP levels are higher in the OR blood on day 21, and in the OCR blood on days 7, 14, 21, and 28 compared to the conventional stored blood (control). Also, the OR blood has higher levels of 2,3-DPG on days 2, 7, and 14 compared to the control. Additionally, the OCR blood shows higher levels of 2,3-DPG on days 2, 7, and 14 compared to the control. Refer to Figure 2.

[0106] Example 2: Recovery Rate of Oxygen-Reduced Blood Label the control, OR, and OCR blood samples (less than 200 μL) stored for 3 weeks with technetium 99m. Transfuse the labeled RBCs (200 μL) into the animals and measure the circulating radioactivity periodically up to 24 hours after transfusion to estimate the percentage of transfused RBCs that are still viable 24 hours after transfusion. As shown in Figure 3, when RBCs are stored for 3 weeks, significantly more OR- and OCR-RBCs are recovered compared to control RBCs.

[0107] Example 3: Rat Model of Hemorrhagic Shock Resuscitation Blood Collection and Sample Preparation: Collect the red blood cells from a total of 12 - 14 rats into CP2D anticoagulant for each pool. Pooled blood is leukoreduced using a neonatal leukoreduction filter, the components are separated, and the RBCs are stored in AS-3 additive solution. A total of 6 pools of RBCs are collected. Two pools are prepared for conventional storage (control). Two pools are depleted of oxygen (oxygen reduction; OR), and the remaining two blood pools are depleted of both oxygen and carbon dioxide (oxygen and carbon dioxide reduction; OCR). Each of the 4 pools to be reduced is processed by transferring the RBCs into a 600 mL PVC transfusion bag and creating the final RBC product by gas exchange treatment. Fill the RBC bag with 100% N2 (for OR) or 95% N2 / 5% CO2 (for OCR) through a sterile filter and gently rotate at 60 - 90 RPM on its long side. After 10 minutes, remove the gas through the filter and introduce fresh gas for subsequent gas exchange treatment. Repeat this process 5 - 8 times until the target SO2% of 5 - 10% is achieved as measured by an ABL-90 CO oximeter (Radiometer Copenhagen). The OR and OCR blood is stored anaerobically in a N2-filled canister.

[0108] Experiments are performed on Sprague-Dawley rats (Charles River Laboratories, Boston, MA) weighing 150 - 200 grams (g). Briefly, the animals are anesthetized by intraperitoneal administration of 40 mg / kg pentobarbital sodium. The animals are placed supine on a heating pad to maintain a deep body temperature of 37°C. For the animals, prepare (i) catheterization of the left jugular vein and the left femoral artery, (ii) tracheotomy (polyethylene-90 tube), and (iii) introduction of a left ventricle (LV) conductance catheter via the right carotid artery. For the animals, mechanical artificial ventilation using room air (TOPO ventilator (Kent Scientific, Torrington, CT)) is performed at a respiratory rate of 50 - 70 breaths per minute and 10 - 15 cmH2O. After measurement, volatile anesthesia (1.5% / vol isoflurane (Dragerwerk AG, Laubeck, Germany)) is administered using a vaporizer connected to the ventilator. The depth of anesthesia is continuously verified by toe pinch, and isoflurane is increased by 0.1% / vol as needed to prevent discomfort in the animals.

[0109] Anesthetized animals are bled by removing 50% of the animal's blood volume within 10 minutes via a femoral artery catheter, placing the animals in a state of hypovolemic shock. The hypovolemic shock state is maintained for 30 minutes. Pre-stored RBCs are infused at a rate of 300 microliters per minute (μL / min) via the femoral artery to perform resuscitation until the mean arterial pressure (MAP) stabilizes at 90% of the baseline during a 60-minute resuscitation period. At 10, 20, 30, 45, and 60 minutes during this period, MAP and heart rate (HR) are obtained from the femoral artery catheter (PowerLab (AD Instruments, Colorado Springs, CO)). After 60 minutes, hematocrit (Hct) is measured via centrifugation of heparinized capillaries. Hemoglobin (Hb), lactate, glucose, K+, Na+, pH, and arterial blood gases are quantified by an ABL90 CO-oximeter (Radiometer, Copenhagen). Cardiac function indices and systemic values (MAP, HR, Hct, Hb, and blood gases) are monitored at baseline (BL), during shock, and at 10 minutes (early R), 20 minutes, 30 minutes, 45 minutes, and 60 minutes (late R) after resuscitation. Animals are euthanized at the end of the experiment.

[0110] Example 4: Hematocrit Analysis in a Rat Model of Hemorrhagic Shock Hematocrit (Hct) is reduced by approximately 30 - 40% after induction of hypovolemic shock. Hematocrit can be restored to normal levels by supplying conventionally stored, OR, or OCR blood stored for 1 week. See Figure 4A. However, as shown in Figure 4B, OR blood stored for 1 week shows an increase in hematocrit percentage compared to control and OCR blood 10 minutes after resuscitation (early R). The hematocrit percentage of OR blood is maintained at an improved level compared to the control 60 minutes after resuscitation (late R).

[0111] Example 5: Changes in Mean Arterial Pressure with Oxygen-Reduced Blood Mean arterial pressure (MAP) was obtained from the femoral artery catheter (PowerLab, AD Instruments, Colorado Springs, CO). As shown in Figure 5A, the baseline MAP was between 80 and 110 mmHg. MAP was reduced to between 20 and 60 mmHg during hemorrhagic shock. By resuscitating the animals with OR and OCR blood stored for 1 week, MAP increased to approximately 80 and 90 mmHg, respectively. As shown in Figure 5B, resuscitation with OR blood was able to restore MAP to the normal range 10 minutes after resuscitation compared to the control. Control and OCR stored blood were able to restore MAP to the normal range 60 minutes after resuscitation. The amount of blood required for resuscitation and maintenance of hemodynamics using conventional stored RBCs (control) was greater than the required OR and OCR RBCs. See Figures 6A and 6B.

[0112] Example 6: Metabolic response to hemorrhagic shock Hemorrhagic shock in animals increases lactate levels from about 2 mmol / L to about 8 - 14 mmol / L. Resuscitation using OR and OCR RBCs stored for 1 week reduces lactate to near normal levels just 10 minutes after resuscitation. See Figure 7A. The lactate levels of animals resuscitated with control blood are similar to those of animals under hemorrhagic shock. Animals treated for 60 minutes with control, OR, and OCR RBCs show similar lactate levels. As shown in Figure 7B, OR RBCs stored for 3 weeks can also reduce lactate levels compared to the control 10 minutes after resuscitation. However, 80 minutes after resuscitation, OCR RBCs restored lactate levels to the normal range. Control and OR RBCs were able to reduce lactate levels but not to the normal range of 1 - 3 mmol / L. Analysis of glucose levels shows that in animals under hemorrhagic shock, glucose in the normal range of about 160 mg / dL to about 240 mg / dL increases to the range of about 320 to about 510 mg / dL. See Figures 8A and 8B. Both OR and OCR RBCs stored for 1 week reduce glucose levels compared to the control 10 minutes after resuscitation. All three samples restored glucose levels to the normal range 60 minutes after resuscitation. As shown in Figure 8B, OR and OR RBCs stored for 3 weeks can also reduce glucose levels compared to the control 10 minutes after resuscitation. Unlike RBCs stored for 1 week, only OR and OCR RBCs were able to restore glucose to within the normal range. Thus, both lactate and glucose levels are reduced more rapidly in OR and OCR RBCs compared to control RBCs.

[0113] Example 7: Injury and Inflammation of Vital Organs Analyze the organ injury and inflammation in animals after experiencing hemorrhagic shock and resuscitation. An increase in the level of liver enzymes indicates some form of liver damage or injury. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were analyzed to quantify liver injury. Resuscitation using OR and OCR RBCs stored for 1 week (Figures 9A and 10A) and OR and OCR RBCs stored for 3 weeks (Figures 9B and 10B) reduced AST and ALT levels compared to control RBCs stored for the same time period. Serum creatinine and blood urea nitrogen (BUN) levels were analyzed to quantify kidney function. OR and OCR RBCs stored for 1 week reduced serum creatinine levels by more than 30% compared to control RBCs (Figure 11A). After 3 weeks of storage, the serum creatinine levels of animals treated with control, OR, and OCR RBCs overlapped (Figure 11B). BUN levels were more than 30% lower in animals treated with OCR RBCs stored for 1 week compared to the control (Figure 12A). Also, BUN levels were more than 30% lower in animals treated with OR RBCs stored for 3 weeks compared to the control (Figure 12B). Overall, the function of vital organs was maintained in the case of OR and OCR RBCs compared to control RBCs.

[0114] At the end of the in vivo test, the liver, spleen, and lungs were excised from the animals, and various inflammatory factors including CXCL1, urinary neutrophil gelatinase-associated lipocalin (u-NGAL), IL-6, and neutrophils were analyzed. CXCL1 was reduced in animals treated with OR and OCR RBCs stored for 1 or 3 weeks compared to controls stored for the same time period (Figures 13A, B, 14A, B, and 15A, B). As shown in Figures 16A and B, u-NGAL was reduced in the kidneys of animals treated with OR and OCR RBCs stored for 1 or 3 weeks compared to control RBCs stored for an equivalent amount of time (Figure 16). As shown in Figures 17 and 18, the percentages regarding the levels of CD45+ neutrophils and IL-6 in the lungs excised from the animals were significantly decreased in OR and OCR RBCs compared to control RBCs stored for the same time period. These results indicate that organ damage and inflammation are reduced in animals treated with OR and OCR RBCs compared to those treated with control RBCs.

Claims

**Claim 1** A method for treating low mean arterial pressure in a subject in need of treatment for low mean arterial pressure, the method comprising supplying oxygen-reduced stored blood to a subject having low mean arterial pressure due to hemorrhagic trauma, wherein the oxygen-reduced blood has an initial oxygen saturation of 20% or less and is maintained at an oxygen saturation of 20% or less during the storage period; said method. **Claim 2** The method according to claim 1, wherein the mean arterial pressure in the subject in need of treatment increases in comparison with a patient who has received conventional stored blood after said supply. **Claim 3** The method according to claim 1, wherein the hemorrhagic trauma is selected from the group consisting of surgery, penetrating trauma, blunt trauma, injury from a fall, and injury from a motor vehicle accident. **Claim 4** The method according to claim 1, wherein the rate of increase of the mean arterial pressure in the subject in need of treatment is faster than the rate of increase of the mean arterial pressure in a subject not receiving the oxygen-reduced stored blood. **Claim 5** The method according to claim 1, wherein the mean arterial pressure in the subject in need of treatment increases by at least 20% in comparison with the mean arterial pressure of a patient who has received a transfusion of conventional stored blood after said supply. **Claim 6** The method according to claim 5, wherein the mean arterial pressure in the subject maintains at least a 20% increase for at least 1 hour after said supply. **Claim 7** A method for reducing the amount of blood required for transfusion in a trauma patient in need of transfusion, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage; said method. **Claim 8** A method for reducing hemorrhagic shock in a trauma patient in need of reduction of hemorrhagic shock, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage, wherein the trauma patient has a lactate level between 0.5 and 2.5 mmol / L before said supply and the hemorrhagic shock returns to normal; said method. **Claim 9** A method for reducing liver injury in a trauma patient in need of blood transfusion therapy, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage; said method. **Claim 10** The method according to claim 9, wherein the trauma patient has an improvement in AST level, ALT level, or a combination thereof after said supply. **Claim 11** A method for reducing renal insufficiency in trauma patients requiring blood transfusion therapy, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

12. The method according to claim 11, wherein the trauma patient has an improvement in a level selected from neutrophil gelatinase-associated lipocalin (NGAL), serum creatinine, blood urea nitrogen (BUN), or a combination thereof.

13. A method for reducing lung injury in trauma patients requiring blood transfusion therapy, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

14. A method for reducing lactic acid in trauma patients requiring reduction of lactic acid, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

15. A method for reducing AST in trauma patients requiring reduction of AST, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

16. A method for reducing ALT in trauma patients requiring reduction of ALT, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

17. A method for reducing BUN in trauma patients requiring reduction of BUN, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

18. A method for reducing neutrophil gelatinase-associated lipocalin (NGAL) in trauma patients requiring reduction of neutrophil gelatinase-associated lipocalin, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

19. A method for reducing serum creatinine in trauma patients requiring reduction of serum creatinine, the method comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

20. Use of donor blood for producing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage for the therapeutic use of low mean arterial pressure.

21. Use of donor blood for producing oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage for therapeutic use in reducing adverse events in patients with hemorrhagic trauma, wherein the adverse events are selected from the group consisting of liver injury, lung failure, kidney failure, and heart failure.

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