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Background Study Design and Methods Results Conclusions The ACP 215 automated cell processor is used to glycerolize and deglycerolize red cell concentrates (RCCs). Its primary advantage over the COBE 2991, previously used to cryopreserve RCCs, is that it maintains a closed system enabling extended post‐thaw expiry. However, it was observed that post‐deglycerolization hematocrits (Hct) of units processed with the LN236 kit are markedly lower than those processed using the COBE 2991. Therefore, we intended to determine whether a modified process using a smaller volume deglycerolization kit (LN235) could increase the final Hct with limited deleterious effects on product characteristics.Two proof‐of‐concept (POC) studies, conducted to determine the feasibility of using the LN235 processing kit for deglycerolization, identified the necessary modifications to the pre‐ and post‐deglycerolization process, after which a two‐part study characterized the modified protocol. The impact of pre‐cryopreservation storage duration (7–21 days), input red cell mass, and the type of CPD/SAGM RCC production method (red cell filtration and whole blood filtration) were investigated.Using the LN235 kit in conjunction with a volume reduction step for RCCs with a red cell mass exceeding 180 mL allowed for an ~8% increase in Hct. As expected, slightly lower recoveries were seen for large RCCs due to volume reduction; however, there were no other detrimental outcomes on product quality.Leveraging the LN235 kit, recommended by Haemonetics for units with a red cell mass of ≤180 mL, can be used to increase the post‐deglycerolization Hct of RCCs that exceed this volume. [ABSTRACT FROM AUTHOR]

BACKGROUND: Ideal blood inventory management involves guaranteeing maximal availability of blood while minimizing wastage. Benchmark for the guidance of O (Rh) D-negative red blood cells (ONEG RBCs) is not widely available. In this study, we aimed to identify the areas of improvement in blood center inventory of ONEG RBCs through a clinical audit. MATERIALS AND METHODS: During April 2017 to March 2018, patients who received ONEG RBCs units were studied for their demographics, primary reason for admission, location, and clinical condition. Data were collected from computerized blood center information system, online integrated laboratory data (Integrated Laboratory Management System), and patients’ medical record charts. Children at ≤18 years were included in the pediatric population as per our institutional criterion while a female between 15 and 49 years was considered as having childbearing potential according to previously published data. RESULTS: Overall, 807 units (2.8%) of ONEG RBCs were transfused during 577 transfusion events with a median (inter quartile range) of 2 (1–3) units per patient in each transfusion event. Recipients of ONEG RBCs were 221 unique patients including 91 females (41%) and 130 males (59%) and only 44 (20%) females had child-bearing potential. Overall, 72 of 807 red cell units (8.9%) were transfused to young females of O/non-O negative/unknown group and were classified as “obligatory.” Neonates, pediatric patients, chronically transfused, and bone marrow transplant recipients received 337 of 807 (42%) units and were marked as “acceptable.” Transfusion of 398/807 units (49%) to females of nonchildbearing potential and adult males could have been saved for those with a mandatory transfusion requirement of ONEG RBCs. CONCLUSIONS: This clinical audit showed that 409 of 807 of ONEG RBCs (51%) were transfused according to the guidelines while 398 of 807 of these (49%) could have been saved for other mandatory requirements. Appropriate policies, planning, education of physicians, and regular clinical audits are needed to bring the desired change in transfusion practices.

BACKGROUND: Ideal blood inventory management involves guaranteeing maximal availability of blood while minimizing wastage. Benchmark for the guidance of O (Rh) D-negative red blood cells (ONEG RBCs) is not widely available. In this study, we aimed to identify the areas of improvement in blood center inventory of ONEG RBCs through a clinical audit. MATERIALS AND METHODS: During April 2017 to March 2018, patients who received ONEG RBCs units were studied for their demographics, primary reason for admission, location, and clinical condition. Data were collected from computerized blood center information system, online integrated laboratory data (Integrated Laboratory Management System), and patients' medical record charts. Children at ≤18 years were included in the pediatric population as per our institutional criterion while a female between 15 and 49 years was considered as having childbearing potential according to previously published data. RESULTS: Overall, 807 units (2.8%) of ONEG RBCs were transfused during 577 transfusion events with a median (inter quartile range) of 2 (1–3) units per patient in each transfusion event. Recipients of ONEG RBCs were 221 unique patients including 91 females (41%) and 130 males (59%) and only 44 (20%) females had child-bearing potential. Overall, 72 of 807 red cell units (8.9%) were transfused to young females of O/non-O negative/unknown group and were classified as "obligatory." Neonates, pediatric patients, chronically transfused, and bone marrow transplant recipients received 337 of 807 (42%) units and were marked as "acceptable." Transfusion of 398/807 units (49%) to females of nonchildbearing potential and adult males could have been saved for those with a mandatory transfusion requirement of ONEG RBCs. CONCLUSIONS: This clinical audit showed that 409 of 807 of ONEG RBCs (51%) were transfused according to the guidelines while 398 of 807 of these (49%) could have been saved for other mandatory requirements. Appropriate policies, planning, education of physicians, and regular clinical audits are needed to bring the desired change in transfusion practices. [ABSTRACT FROM AUTHOR]

Background: There are scientific evidence confirmed specific changes in blood cell counts, reducing the efficacy and feasibly the safeness of blood transmission when storing blood at 4°C for 5 weeks or more. Objectives: The study aimed to investigate the effects of stored blood obtained from healthy subjects and patients with anemia due to iron deficiency and beta-thalassemia, on hematological indices. Materials and Methods: A total of 37 participants, consisting of 14 healthy subjects, 13 patients with iron-deficiency anemia, and 10 patients with beta-thalassemia minor, were recruited from Hiwa Hospital between November 2021 and July 2022. Blood samples were obtained from the participants and stored at 4°C for 5 weeks. Hematological indices, including red cell distribution, platelet distribution width, and mean platelet volume, were determined using a hematology analyzer at weekly intervals. Results: Blood storage caused significantly increased mean values of hematological indices among healthy subjects as well as among patients with iron-deficiency anemia and beta-thalassemia, although the pattern of changes was differed. Conclusions: The storage of whole blood significantly increased hematological indices, showing variations in both healthy subjects and patients with iron-deficiency anemia and beta-thalassemia. The pattern of raise in these hematological indices is specific to iron-deficiency anemia and thalassemia when compared with healthy subjects. [ABSTRACT FROM AUTHOR]

Xenon, an inert gas commonly used in medicine, has been considered as a potential option for prolonged preservation of donor packed red blood cells (pRBCs) under hypoxic conditions. This study aimed to investigate how xenon affects erythrocyte parameters under prolonged storage. In vitro model experiments were performed using two methods to create hypoxic conditions. In the first method, xenon was introduced into bags of pRBCs which were then stored for 42 days, while in the second method, xenon was added to samples in glass tubes. The results of our experiment showed that the presence of xenon resulted in notable alterations in erythrocyte morphology, similar to those observed under standard storage conditions. For pRBC bags, hemolysis during storage with xenon exceeded the acceptable limit by a factor of six, whereas the closed-glass-tube experiment showed minimal hemolysis in samples exposed to xenon. Notably, the production of deoxyhemoglobin was specific to xenon exposure in both cell suspension and hemolysate. However, this study did not provide evidence for the purported protective properties of xenon. [ABSTRACT FROM AUTHOR]

The recently officially approved and released health industry standard Standard for Blood Storage(WS 399-2023) by the National Health Commission is the latest revised version of Standard for Blood Storage(WS 399-2012), which mainly stipulates the storage requirements for whole blood, red blood cells, platelets, granulocytes, plasma and irradiated blood. It applies to blood collection and supply institutions and healthcare institutions, and is the guideline for the safe use of blood.

Blood transfusion, one of them, is a process of transferring blood components containing glucose, lactate, and potassium. During storage, blood cells undergo metabolic changes such as decreased blood pH, hemolysis, and increased potassium levels. One of the complications of transfusion that must be avoided is hyperkalemia due to the release of potassium. This study aimed to determine differences in stored blood potassium levels and PRC QC results during 11 days of storage. This was an analytic observational study with a cross sectional approach. A T-test was used to analyze the difference. In addition, PRC blood quality control (QC) was also carried out for stored blood according to standards. This study used 40 samples of fresh blood bags obtained from UTD PMI Yogyakarta City. The mean potassium level in fresh blood bags was 4.373 ± 0.417 mmol/L (3.70-5.50) and the average potassium level in blood bags stored for 6 days was 8.290 ± 0.275 mmol/L (7.80- 8.90). There was a significant difference in potassium levels between fresh blood bags and blood bags stored for 6 days (p = 0.000). The overall QC PRC result is 50% which indicates the need for improvement in terms of the blood component separation process. [ABSTRACT FROM AUTHOR]

BackgroundHemoglobin (Hb)-based oxygen carriers (HBOCs) have been proposed as alternatives to blood for decades. Previous studies demonstrated that large molecular diameter HBOCs based on polymerized bovine Hb (PolybHb) attenuate Hb side-effects and toxicity. The objective of this study was to test the safety and efficacy of tense state PolybHb after long-term storage.Methods and resultsPolybHb was subjected to diafiltration to remove low molecular weight (< 500 kDa) species and stored for 2 years. PolybHb was studied in parallel with blood, collected from rats and stored leukodepleted under blood bank conditions for 3 weeks. Rats were hemorrhaged and resuscitated to 90% of the blood pressure before the hemorrhage with fresh blood, stored blood, fresh PolybHb, or 2-year-stored PolybHb. Hemorrhagic shock impaired oxygen delivery and cardiac function. Resuscitation restored blood pressure and cardiac function, but stored blood required a significantly larger transfusion volume to recover from shock compared with fresh blood and PolybHb (fresh and stored). Stored blood transfusion elevated markers of organ damage compared with all other groups.ConclusionsThese studies indicate that large molecular diameter PolybHb is as efficacious as fresh blood in restoring cardiac function and confirm the lack of degradation of PolybHb's safety or efficacy during long-term storage.

BackgroundResuscitation from hemorrhagic shock (HS) by blood transfusion restores oxygen (O2) delivery and provides hemodynamic stability. Current regulations allow red blood cells (RBCs) to be stored and used for up to 42 days. During storage, RBCs undergo many structural and functional changes. These storage lesions have been associated with adverse events and increased mortality after transfusion, increasing the need for improved RBC storage protocols. This study evaluates the efficacy of anaerobically stored RBCs to resuscitate rats from severe HS compared with conventionally stored RBCs.Methods and resultsRat RBCs were stored under anaerobic, anaerobic/hypercapnic, or conventional conditions for a period of 3 weeks. Hemorrhage was induced by controlled bleeding, shock was maintained for 30 min, and RBCs were transfused to restore and maintain blood pressure near the prhemorrhage level. All storage conditions met current regulatory 24-h posttransfusion recovery requirements. Transfusion of anaerobically stored RBCs required significantly less RBC volume to restore and maintain hemodynamics. Anaerobic or anaerobic/hypercapnic RBCs restored hemodynamics better than conventionally stored RBCs. Resuscitation with conventionally stored RBCs impaired indices of left ventricular cardiac function, increased hypoxic tissue staining and inflammatory markers, and affected organ function compared with anaerobically stored RBCs.ConclusionsResuscitation from HS via transfusion of anaerobically stored RBCs recovered cardiac function, restored hemodynamic stability, and improved outcomes.

Blood samples are routinely collected from wild populations in remote locations with limited electricity, minimal diagnostic capabilities, and extreme environmental conditions. Under these conditions, serum samples may be stored for prolonged time under varied temperatures prior to processing, which could affect the ability to interpretation the results. This study’s objective was to evaluate the effects of delayed processing of serum samples and varied storage temperatures on biochemical values in African savanna elephants (Loxodonta africana). Blood samples were collected from six elephants managed by the North Carolina Zoo. For each elephant, seven red top tubes were collected. One serum sample for each elephant was analyzed on Day 0 (control group). The remaining samples were stored under different temperatures including room temperature (23 °C), refrigeration (2.2 °C), and incubation (32.2 °C), with samples from each temperature group being analyzed on Day 5 and Day 10. Many of analytes (10 out of 18) did not change significantly regardless of storage temperature or time. Refrigeration improved stability in an additional four analytes over prolonged storage. We conclude that if serum is properly separated shortly after collection, many serum biochemical analytes can be accurately measured even after suboptimal serum storage, but refrigeration and prompt evaluation are still required for some analytes.

Background and Objectives: Stored blood undergoes many metabolic, biochemical, and molecular changes known as storage lesions which are responsible for negative posttransfusion effects in recipients. This study aims to detect serial in vitro qualitative changes that occur during processing and blood storage including changes in plasma hemoglobin (Hb), plasma lactate dehydrogenase (LDH), and potassium levels and changes (fall) in pH of packed red blood cell at serial intervals under a standard condition of storage in the blood bank. This is a two year prospective study. Methods: Fifty donors were selected and blood units were collected and stored under blood bank conditions. Mean plasma Hb of stored blood was estimated by measuring the transmittance and absorbance of modified azide-methemoglobin and percentage hemolysis was calculated on days 0, 1, 7, 21, 28, 35, and 42 days. Similarly, plasma LDH and plasma potassium levels were also assessed during storage. Results: It was noted that free Hb level and percentage hemolysis progressively increased with storage along with the level of LDH and potassium. However, the extent of hemolysis did not exceed the permissible limit of up to 42 days of storage. Conclusion: It was concluded that quantitative estimation of Hb is superior to the visual method for the determination of hemolysis. Hemolysis can be further reduced by additive solution saline, adenine, glucose, and mannitol and using di-2-ethyl hexyl pHthalate as s plasticizer in blood bags for storage.

Background: Citrate is the only anticoagulant currently Food and Drug Administration (FDA)‐approved for the long‐term storage of blood for transfusion. Citrate inhibits phosphofructokinase and may play a pro‐inflammatory role, suggesting that there may be an advantage to using alternative anticoagulants. Here, we examine the use of pyrophosphate as an anticoagulant. Study design and methods: Whole blood samples from healthy donors were anticoagulated either with citrate–phosphate–adenine–dextrose (CPDA‐1) or our novel anticoagulant mixture pyrophosphate‐phosphate–adenine–dextrose (PPDA‐1). Samples were assessed for coagulation capacity by thromboelastography immediately after anticoagulation (T0) with and without recalcification, as well as 5 hours after anticoagulation (T1) with recalcification. Complete blood counts were taken at both timepoints. Flow cytometry to evaluate platelet activation as well as blood smears to evaluate cellular morphology were performed at T1. Results: No clotting was detected in samples anticoagulated with either solution without recalcification. After recalcification, clotting function was restored in both groups. R‐Time in recalcified PPDA‐1 samples was shorter than in CPDA‐1 samples. A reduction in platelet count at T1 compared to T0 was observed in both groups. No significant platelet activation was observed in either group at T1. Blood smear indicated platelet clumping in PPDA‐1. Conclusion: We have shown initial proof of concept that pyrophosphate functions as an anticoagulant at the dose used in this study, though there is an associated loss of platelets over time that may limit its usefulness for blood storage. Further dose optimization of pyrophosphate may limit or reduce the loss of platelets. [ABSTRACT FROM AUTHOR]

Background and Objectives: Stored blood undergoes many metabolic, biochemical, and molecular changes known as storage lesions which are responsible for negative posttransfusion effects in recipients. This study aims to detect serial in vitro qualitative changes that occur during processing and blood storage including changes in plasma hemoglobin (Hb), plasma lactate dehydrogenase (LDH), and potassium levels and changes (fall) in pH of packed red blood cell at serial intervals under a standard condition of storage in the blood bank. This is a two year prospective study. Methods: Fifty donors were selected and blood units were collected and stored under blood bank conditions. Mean plasma Hb of stored blood was estimated by measuring the transmittance and absorbance of modified azide-methemoglobin and percentage hemolysis was calculated on days 0, 1, 7, 21, 28, 35, and 42 days. Similarly, plasma LDH and plasma potassium levels were also assessed during storage. Results: It was noted that free Hb level and percentage hemolysis progressively increased with storage along with the level of LDH and potassium. However, the extent of hemolysis did not exceed the permissible limit of up to 42 days of storage. Conclusion: It was concluded that quantitative estimation of Hb is superior to the visual method for the determination of hemolysis. Hemolysis can be further reduced by additive solution saline, adenine, glucose, and mannitol and using di-2-ethyl hexyl pHthalate as s plasticizer in blood bags for storage. [ABSTRACT FROM AUTHOR]

The hypothesis of the potential impact of the sex of red blood cell (RBC) concentrate (RCC) donors, as well as the sex of the recipients, on the clinical outcome, is still under evaluation. Here, we have evaluated the sex impact on RBC properties using in vitro transfusion models. Using a "flask model", RBCs from RCCs (representing the donor)—at different storage lengths—were incubated in a sex-matched and sex-mismatched manner with fresh frozen plasma pools (representing the recipient) at 37 °C, with 5% of CO2 up to 48 h. Standard blood parameters, hemolysis, intracellular ATP, extracellular glucose and lactate were quantified during incubation. Additionally, a "plate model", coupling hemolysis analysis and morphological study, was carried out in similar conditions in 96-well plates. In both models, RBCs from both sexes hemolyzed significantly less in female-derived plasma. No metabolic or morphological differences were observed between sex-matched and -mismatched conditions, even though ATP was higher in female-derived RBCs during incubations. Female plasma reduced hemolysis of female- as well as male-derived RBCs, which may be related to a sex-dependent plasma composition and/or sex-related intrinsic RBC properties. [ABSTRACT FROM AUTHOR]

Background: The risk of military and civilian radiation exposure is increasing, and determining the effects of exposure is a high priority. Irradiation of the nearby blood supply after a nuclear event may impede mobilization of blood products for resuscitation at a time of great need. RBCs are administered to patients with trauma and hemorrhage to transport and deliver oxygen and avoid tissue hypoxia. Here we determine the effects of ionizing radiation on the energy metabolome of RBCs isolated from cold stored whole blood to determine if their stability is compromised by radiation exposure. Study Design and Methods: Whole blood from healthy volunteers was subjected to 0, 25, or 75 Gy of X‐irradiation, and stored at 4°C. RBCs were isolated from stored WB at 0, 1, 7, 14, and 21 days of storage. The levels of extracted Krebs cycle intermediates, nicotinamide adenine dinucleotides, and phosphorylated derivatives of adenosine and guanosine were determined by tandem mass spectroscopy. Results: Irradiation at either 25Gy or 75Gy had no significant effect on any parameter measured compared to control (0Gy). However, there was a significant change over time in storage for ATP, GDP, and guanosine. Discussion: Irradiation at doses up to 75Gy had no effect on the energy metabolome of RBCs prepared from blood stored at 4°C for up to 21 days, suggesting that the RBC energy metabolome is not affected by radiation exposure and the blood can still be used for resuscitation in trauma patients. [ABSTRACT FROM AUTHOR]

Background: Pathogen reduction technology (PRT) may improve the safety of RBCs for transfusion. As the Czech Republic considers PRT, we asked what effects riboflavin and UV light PRT pre‐freezing has on the post‐thaw recovery and properties of cryopreserved RBCs (CRBCs) after deglycerolization and liquid storage. Study Design and Methods: 24 Group O whole blood (WB) units were leukoreduced and then treated with riboflavin and UV light PRT (Mirasol, Terumo BCT, USA) before cryopreservation (T‐CRBC); 20 similarly‐collected units were untreated controls (C‐CRBC). Units were processed to RBCs and then cryopreserved with 40% glycerol (wt/vol), frozen at −80°C, stored >118 days, reconstituted as deglycerolized RBC units in AS‐3, and stored at 4 ± 2°C for 21 days. One treated unit sustained massive hemolysis during the post‐thaw wash process and was removed from data analysis. The remaining units were assessed pre‐PRT, post‐PRT, and post‐thaw‐wash on days 0, 7, 14, and 21 for hematocrit, volume, hemoglobin per transfusion unit, pH, % hemolysis, hemoglobin in the supernatant, potassium, phosphorus, NH3, osmolality, ATP, and 2,3‐diphosphoglycerate. Results: PRT with leukoreduction caused a 5% loss of RBC followed by a 24% freeze–thaw‐wash related loss for a total 28% loss but treated units contained an average of 45 g of hemoglobin, meeting European Union guidelines for CRBC. T‐CRBCs displayed higher post‐wash hemolysis, potassium, and ammonia concentrations, and lower ATP at the end of storage. Conclusions: Cryopreserved RBCs from Riboflavin and UV light‐treated WB meet the criteria for clinical use for 7 days after thawing and provide additional protection against infectious threats. [ABSTRACT FROM AUTHOR]

Background: Omics technologies represent a new analytical approach that allows a full cellular readout through the simultaneous analysis of thousands of molecules. The application of such technologies represents a flourishing field of research in human medicine, especially in transfusion medicine, while their application in veterinary medicine still needs to be developed. Summary: Omics technologies, especially proteomics, metabolomics, and lipidomics, are currently applied in several fields of human medicine. In transfusion medicine, the creation and integration of multiomics datasets have uncovered intricate molecular pathways occurring within blood bags during storage. In particular, the research has been directed toward the study of storage lesions (SLs), i.e., those biochemical and structural changes that red blood cells (RBCs) undergo during hypothermic storage, their causes, and the development of new strategies to prevent them. However, due to their challenges to perform and high costs, these technologies are hardly accessible to veterinary research, where their application dates back only to the last few years and thus a great deal of progress still needs to be made. As regards veterinary medicine, there are only a few studies that have focused mainly on fields such as oncology, nutrition, cardiology, and nephrology. Other studies have suggested omics datasets that provide important insights for future comparative investigations between human and nonhuman species. Regarding the study of storage lesions and, more generally, the veterinary transfusion field, there is a marked lack of available omics data and results with relevance for clinical practice. Key Messages: The use of omics technologies in human medicine is well established and has led to promising results in blood transfusion and related practices knowledge. Transfusion practice is a burgeoning field in veterinary medicine, but, to date, there are no species-specific procedures and techniques for the collection and storage of blood units and those validated in the human species are univocally pursued. Multiomics analysis of the species-specific RBCs' biological characteristics could provide promising results both from a comparative perspective, by increasing our understanding of species suitable to be used as animal models, and in a strictly veterinary view, by contributing to the development of animal-targeted procedures. [ABSTRACT FROM AUTHOR]

Introduction and aim. Stored erythrocytes develop lesions involving changes in their structure and function reducing their efficacy. Oxidative Stress (OS) being one of the main causes of storage lesion, can be attenuated by antioxidants as additives in the storage solution. This study aims to evaluate the effect of p-Coumaric acid (CA) on erythrocytes during whole blood storage. Material and methods. Blood collected from Male Wistar rats was stored at 4°C in CPDA-1 solution for 21 days. Blood samples were stored with and without 1mM CA (CA 1) and 10 mM CA (CA 10). The erythrocytes were isolated every week during storage and the biomarkers for OS and antioxidant status were analysed. Results. Superoxide dismutase and catalase elevated on day 14. Conjugate dienes decreased in CA 10 on day 14. Thiobarbituric acid reactive substances increased on day 7 and decreased on day 14 in CA groups. Protein sulfhydryls decreased in controls and CA 1 on day 14 whereas, it was maintained in CA 10. Conclusion. Coumaric acid upregulated the antioxidant enzymes and protected the cells from oxidative damage. Thus, coumaric acid can be employed as a potent additive during storage and opens new avenues of employing it in similar OS situations in erythrocytes. [ABSTRACT FROM AUTHOR]

Introduction: The impact of blood storage on red blood cell (RBC) alloimmunization remains controversial, with some studies suggesting enhancement of RBC‐induced alloantibody production and others failing to observe any impact of storage on alloantibody formation. Since evaluation of storage on RBC alloimmunization in patients has examined antibody formation against a broad range of alloantigens, it remains possible that different clinical outcomes reflect a variable impact of storage on alloimmunization to specific antigens. Methods: RBCs expressing two distinct model antigens, HEL‐OVA‐Duffy (HOD) and KEL, separately or together (HOD × KEL), were stored for 0, 8, or 14 days, followed by detection of antigen levels prior to transfusion. Transfused donor RBC survival was assessed within 24 h of transfusion, while IgM and IgG antibody production were assessed 5 and 14 days after transfusion. Results: Stored HOD or KEL RBCs retained similar HEL or KEL antigen levels, respectively, as fresh RBCs, but did exhibit enhanced RBC clearance with increased storage age. Storage enhanced IgG antibody formation against HOD, while the oppositive outcome occurred following transfusion of stored KEL RBCs. The distinct impact of storage on HOD or KEL alloimmunization did not appear to reflect intrinsic differences between HOD or KEL RBCs, as transfusion of stored HOD × KEL RBCs resulted in increased IgG anti‐HOD antibody development and reduced IgG anti‐KEL antibody formation. Conclusions: These data demonstrate a dichotomous impact of storage on immunization to distinct RBC antigens, offering a possible explanation for inconsistent clinical experience and the need for additional studies on the relationship between RBC storage and alloimmunization. [ABSTRACT FROM AUTHOR]

Blood samples are routinely collected from wild populations in remote locations with limited electricity, minimal diagnostic capabilities, and extreme environmental conditions. Under these conditions, serum samples may be stored for prolonged time under varied temperatures prior to processing, which could affect the ability to interpretation the results. This study's objective was to evaluate the effects of delayed processing of serum samples and varied storage temperatures on biochemical values in African savanna elephants (Loxodonta africana). Blood samples were collected from six elephants managed by the North Carolina Zoo. For each elephant, seven red top tubes were collected. One serum sample for each elephant was analyzed on Day 0 (control group). The remaining samples were stored under different temperatures including room temperature (23 °C), refrigeration (2.2 °C), and incubation (32.2 °C), with samples from each temperature group being analyzed on Day 5 and Day 10. Many of analytes (10 out of 18) did not change significantly regardless of storage temperature or time. Refrigeration improved stability in an additional four analytes over prolonged storage. We conclude that if serum is properly separated shortly after collection, many serum biochemical analytes can be accurately measured even after suboptimal serum storage, but refrigeration and prompt evaluation are still required for some analytes. [ABSTRACT FROM AUTHOR]

Abstract Blood transfusions are mainly given to intensive care patients; therefore, additional complications that could arise from storage lesions in preserved blood should be avoided. It has been shown that human stored red blood cells are subject to changes that are considered to be a number of interdependent processes involving metabolic disarrangement and oxidative stress. The aim of our study was to determine alterations in selected hematological and biochemical parameters and to assess whether and when oxidative stress is a significant phenomenon in stored dog CPDA-1 whole blood. Ten ½ unit bags of whole blood donated from dogs and preserved with CPDA-1 (anticoagulant containing citrate, phosphate, dextrose and adenine) were stored for 5 weeks. Each week, a 9 ml sample was drawn aseptically to measure hematological parameters, selected metabolites, free hemoglobin content, osmotic fragility, antioxidant enzyme activity, total antioxidant capacity, malondialdehyde concentration and protein carbonyl content. The results revealed an MCV decrease in the first week of storage and then a gradual increase; osmotic fragility decreased at that time and remained low throughout the study period. Leukodepletion became significant in the fourth week of storage. The free hemoglobin concentration continuously increased, with the greatest changes observed in the last two weeks of storage. The total antioxidant capacity changed in a reverse manner. Superoxide dismutase and glutathione peroxidase activities decreased from week 0 to week 3, and catalase activity tended to decrease over time. The highest malondialdehyde concentrations in blood supernatant were measured in the first week of storage, and the carbonyl concentration increased after 35 days. Hematological changes and oxidative stress are already present in the first week of storage, resulting in depletion of the antioxidant system and subsequent accumulation of oxidation products as well as erythrocyte hemolysis, which are most pronounced at the end of the storage period.

Objective To explore the main development directions and research progress of blood transfusion in the U. S.military based on ClinicalTrials.gov. Methods The literature of clinical trials related to field transfusion medicine from 2002 to 2021 were retrieved based on www.clinicaltrials.gov with "United States Department of Defense" , "U.S.Army Medical Research and Development Command" and "United States Naval Medical Research Center" as the search terms, and the main characteristics were analyzed in terms of research progress, clinical stage, task allocation, regional distribution, research contents and results. Results A total of 931 clinical trials were retrieved, of which 16 were closely related to field blood transfusion medicine.Among them, pathogen reduction/inactivation of whole blood, platelet transfusion, plasma transfusion, whole blood transfusion and other types of researches accounted for 25% (4/16), 25% (4/16), 25% (4/16), 18.75 (3/16) and 6.25% (1/16), respectively.The U. S.military focused on the new technological breakthroughs in blood safety and blood storage, the evaluation of the countershock therapy, the application of whole blood early transfusion and the civil-military inosculation in research. Conclusion This study, based on ClinicalTrials.gov data, redounds to understand the research progress and current situation of the U. S.military field blood transfusion medicine.

The stability of gene transcripts associated with the presence of circulating tumor cells (CTCs) has been predominantly studied in cultured cancer cell lines added to blood samples under artificial conditions. In the present study the effect of storage on CTC-related transcripts was assessed in blood samples taken from patients with non-small lung cancer (n=58). The blood samples were split in two equal parts to compare the gene expression with and without storage for 24 h at ambient temperature without preservative added. After enrichment using the microfluidic Parsortix® technology, the expression levels of selected genes were assessed using quantitative PCR following a gene-specific pre-amplification. The prognostic relevance of each gene in fresh and stored blood samples was evaluated using the R-package Survminer. Some genes were either not affected (TWIST1, CDH5, CK19) or upregulated upon storage (NANOG, MET, UCHL1) but still associated with poor prognosis. In contrast, ERBB3, PTHLH, EpCAM, and TERT were no longer associated with the overall survival of the patients. The study demonstrates the surprising stability of CTC-related transcripts, which makes overnight shipping of native blood samples possible. Careful verification is required when using model systems – such as normal blood spiked with tumor cells – or other CTC-related markers, as individual transcripts may respond differently to storage. [ABSTRACT FROM AUTHOR]

Background and Objectives: In Japan, there are various opinions on the pros and cons of home transfusion because of safety concerns. We hence aimed to elucidate the safety and availability of home transfusion in Japan, which has not been clarified to date. Materials and Methods: Clinics throughout Japan that provide home care and have experience in performing blood transfusions were surveyed. The analysis period was February to December 2019. Basic information about the clinics, their collaboration system with core hospitals, storage method of red blood cells (RBCs) and the system for the management of patient information regarding transfusion reactions were investigated. Results: Detailed information was obtained regarding the implementation of home transfusions by 51 clinics. The proportion of home care clinics performing home transfusions was 17.6%, and they were more frequently performed in urban regions. Approximately half of the clinics collaborated with a core hospital for emergency responses to transfusion reactions. At 84% of the clinics, RBC units were stored in refrigerators that were not exclusively allocated to blood storage. Nurses and family members were involved as patient attendants in 83% and 77% of the home transfusions, respectively. No serious transfusion reactions were reported among the 150 patients in 2019, nor the 623 patients up to 2018. Conclusion: From data on its availability and safety, home transfusions are considered to be in the developing phase in Japan. Increased cooperation between hospitals and clinics is crucial towards improving the home transfusion system in Japan in the future. [ABSTRACT FROM AUTHOR]

Background: Differences in manufacturing conditions using the Haemonetics ACP 215 cell processor result in cryopreserved red cell concentrates (RCCs) of varying quality. This work studied the effect of processing method, additive solution, and storage duration on RCC quality to identify an optimal protocol for the manufacture of cryopreserved RCCs. Materials and methods RCCs were pooled‐and‐split and stored for 7, 14, or 21 days before cryopreservation. Units were glycerolized with the ACP 215 using a single or double centrifugation method. After thawing, the RCCs were deglycerolized, suspended in AS‐3, SAGM, ESOL, or SOLX/AS‐7, and stored for 0, 3, 7, 14, or 21 days before quality testing. Quality assessments included hemoglobin content, hematocrit, hemolysis, adenosine triphosphate (ATP), supernatant potassium, and mean cell volume. Results: Both glycerolization methods produced RCCs that met regulatory standards for blood quality. Dual centrifugation resulted in higher hemoglobin content, fewer processing alerts, and a shorter deglycerolization time than single centrifugation processing. Units processed with AS‐3 and ESOL met regulatory standards when stored for up to 21 days pre‐cryopreservation and 21 days post‐deglycerolization. However, ESOL demonstrated superior maintenance of ATP over RBCs in AS‐3. Some RCCs suspended in SAGM and SOLX exceeded acceptable hemolysis values after 7 days of post‐deglycerolization storage regardless of pre‐processing storage length. Conclusions: When manufacturing cryopreserved RCCs using the ACP 215, dual centrifugation processing with AS‐3 or ESOL additive solutions is preferred, with storage periods of up to 21 days both pre‐processing and post‐deglycerolization. [ABSTRACT FROM AUTHOR]

Oxidative damage by free radicals has a negative effect on blood quality during storage. Antioxidant nanoparticles can prevent oxidative stress. We use SOD‐CAT‐Alb‐PEG‐PLGA‐ nanoparticles to reduce the effects of oxidative stress in blood storage. Electrospray was employed to prepare nanoparticles. Nanoparticles entered the test bags and were kept for 35 days from the time of donation under standard conditions. On target days, experiments were performed on the samples taken. The examination included blood smear, red blood cells count, hemoglobin, hematocrit, K, Fe, glutathione peroxidase, glutathion reductase, glucose‐6‐phosphate dehydrogenase, prooxidant‐antioxidant balance, malondialdehyde, and flow cytometric assay for phosphatidylserine. The repeated measures analysis was performed on samples every week. Morphological changes were less in the test group compared to the control. The quantitative hemolysis profile test showed significant changes in the test and control groups (p < 0.05) in consecutive weeks except for K and Fe. Oxidative stress parameters too showed a significant change during the target days of the examination (p < 0.05). Also, the phosphatidylserine expression was increased in control groups more than test in consecutive weeks (p < 0.05). It seems that the use of antioxidant nanoparticles improves the quality of stored red blood cells and can prevent posttransfusion complications and blood loss by reducing oxidative stress. [ABSTRACT FROM AUTHOR]

Blood transfusions are mainly given to intensive care patients; therefore, additional complications that could arise from storage lesions in preserved blood should be avoided. It has been shown that human stored red blood cells are subject to changes that are considered to be a number of interdependent processes involving metabolic disarrangement and oxidative stress. The aim of our study was to determine alterations in selected hematological and biochemical parameters and to assess whether and when oxidative stress is a significant phenomenon in stored dog CPDA-1 whole blood. Ten ½ unit bags of whole blood donated from dogs and preserved with CPDA-1 (anticoagulant containing citrate, phosphate, dextrose and adenine) were stored for 5 weeks. Each week, a 9 ml sample was drawn aseptically to measure hematological parameters, selected metabolites, free hemoglobin content, osmotic fragility, antioxidant enzyme activity, total antioxidant capacity, malondialdehyde concentration and protein carbonyl content. The results revealed an MCV decrease in the first week of storage and then a gradual increase; osmotic fragility decreased at that time and remained low throughout the study period. Leukodepletion became significant in the fourth week of storage. The free hemoglobin concentration continuously increased, with the greatest changes observed in the last two weeks of storage. The total antioxidant capacity changed in a reverse manner. Superoxide dismutase and glutathione peroxidase activities decreased from week 0 to week 3, and catalase activity tended to decrease over time. The highest malondialdehyde concentrations in blood supernatant were measured in the first week of storage, and the carbonyl concentration increased after 35 days. Hematological changes and oxidative stress are already present in the first week of storage, resulting in depletion of the antioxidant system and subsequent accumulation of oxidation products as well as erythrocyte hemolysis, which are most pronounced at the end of the storage period. [ABSTRACT FROM AUTHOR]

Background and Objectives: Di(2‐ethylhexyl) phthalate (DEHP) must be removed from blood bag sets in Europe by 27 May 2025. DEHP is known to interact with the red blood cell (RBC) membrane, resulting in reduced haemolysis and thus prolonging shelf‐life. Current non‐DEHP alternatives result in increased haemolysis requiring reconsideration of the RBC shelf‐life. Although the immediate impact of eliminating DEHP is to the European community, the non‐DEHP movement could affect blood bag set availability globally. The purpose of this survey is to understand blood centre readiness regarding the transition to non‐DEHP blood collection and storage systems. Materials and Methods: A 24‐question on‐line survey was completed by members of the Biomedical Excellence for Safer Transfusion Collaborative research network. Results: Responses were obtained from 16 blood collection or processing institutions. A majority of respondents (12/16) indicated that both shelf‐life and haemolysis were equally important in selecting non‐DEHP blood bag sets. Six respondents would accept a lower RBC product shelf‐life compared to current practice. Respondents were not clear on the best non‐DEHP vinyl material or RBC storage solution. Three European blood centres indicated they have developed non‐DEHP transition plans. One challenge identified regarding the transition to non‐DEHP is the extensive validation testing that will be required. Conclusion: Blood centres in Europe are concerned with meeting the sunset date for DEHP, considering that limited non‐DEHP blood bag and RBC storage solutions are currently available. Banning DEHP in Europe, which may have global ramifications, represents a major challenge not yet fully understood by the transfusion medicine community. [ABSTRACT FROM AUTHOR]

BackgroundSmall rodent models are routinely used to evaluate the safety and efficacy of blood transfusions. Limited comprehensive literature exists about effect of different storage solutions in rat red blood cells (RBCs) characteristics. RBCs undergo time dependent biochemical and biophysical changes during storage known as hypothermic storage lesions (HSLs).ObjectiveThis study evaluates the effects of RBC additive solutions (AS) during storage of rat RBCs.MethodsBlood was leukoreduced and stored as per manufacturer instructions at 4°C up to 42-days. Three solutions, CPDA-1; AS-1; and AS-7 (SOLX), were evaluated. Biochemical parameters measured included extracellular K+, pH, hemolysis, 2,3-diphosphoglycerate (2,3-DPG), oxygen affinity, ATP, and lactate. Mechanical properties measured included RBC deformability, elongation index (EI), RBC membrane shear elastic modulus (SEM), mean corpuscular volume (MCV), viscosity, and aggregability.ResultsThere were no differences in biochemical or mechanical parameters at baseline or after one week of storage. However, after two weeks, AS-7 preserved biochemical and mechanical properties as compared to CPDA-1 and AS-1. Changes were observed to be significant after 14-days of storage. AS-7 prevented extracellular K+ increase, reduced acidosis, showed lower hemolysis, preserved ATP and 2,3-DPG levels (consequently oxygen affinity), and reduced lactate. AS-7, when compared to CPDA-1 and AS-1, prevented the reduction in RBC deformability and was found to preserve the EI at multiple shear stresses, the membrane SEM, the aggregability and viscosity.DiscussionRat RBCs stored with AS-7 presented reduced changes in biochemical and mechanical parameters, when compared with rat RBCs stored in CPDA-1 and AS-1, after as early as two weeks of storage.

INTRODUCTION: Gamma Irradiation of blood products accentuates biochemical changes in the blood stored at 4°C. This study tried to compare the changes in potassium, sodium, glucose, lactate, and lactate dehydrogenase (LDH) levels in packed red blood cell (PRBC) units irradiated at various time points and then stored versus those stored for a particular period then irradiated.METHODOLOGY: One hundred and eighty units of RBCs were randomly assigned equally to be irradiated or not. Eighteen units each were irradiated by gamma irradiator using cobalt 60 (BI 2000) on day 1, 7, 14, 21, and 28 of their storage, respectively, in the irradiation group. All the units were assessed for their plasma levels of potassium, sodium, glucose, LDH, and lactate by clinical chemistry auto analyzer Beckman coulter AU680 weekly. The values were documented and analyzed by SPSS.RESULTS: Baseline values on day 1 for studied biochemical parameters were comparable between irradiated and nonirradiated groups. Mean values of potassium, lactate and LDH were higher in irradiated than nonirradiated PRBC bags. In contrast, Sodium and Glucose mean values were lower than baseline values. Maximum cumulative mean values were noted in day-21 irradiated bags when the parameters were measured on day-28 for potassium and lactate levels. This was followed by day 14 irradiated bags, followed by day 7 irradiated bags.CONCLUSION: The study indicates that irradiation of red cells later in their storage period had comparatively more detrimental changes in relation to potassium and lactate than irradiation in earlier days. Consideration of irradiation to be performed as close to the issue as possible to reduce a lesser number of days of storage postirradiation is to be explored.

Hemoglobin (Hb) inside and outside the red blood cells (RBCs) undergoes constant transformation to an oxidized form in a process known as autoxidation. The ferrous heme iron (Fe2+) of the prosthetic group is spontaneously transformed into an oxidized ferric (Fe3+) form, but under oxidative stress conditions a higher oxidation ferryl heme (Fe4+) is also formed. Although Fe3+ is a non-functional form of Hb, the Fe4+ is also extremely reactive towards other biological molecules due to its high redox potential. The RBC contains an effective reductive machinery that maintains Hb in the functional form with little oxidation during its life span. The redox transformation of Hb occurs to a lesser extent in young RBCs; it may, however, have detrimental effects on the integrity of these cells during ex vivo storage or when RBCs are subjected to pathogen reduction processes. In this review, Hb oxidation reactions ("oxidative lesion") will be described, including details of how these reactions might impact the clinical use of stored or processed blood for therapeutic purposes. [ABSTRACT FROM AUTHOR]

Introduction: Erythrocytes have flexible, non-nucleated bi-concave shape with lipid bilayer cytoskeleton. Any alterations of erythrocyte shape make it susceptible to hemolysis. Blood for transfusion purpose is routinely stored in Citrate Phosphate Dextrose Adenine (CPDA-1) containing blood bags. During storage, blood undergoes an array of different morphological changes termed as “storage lesions” which makes it more fragile. This study was aimed to determine the structural and functional modifications in erythrocytes in CPDA-1 blood stored in local blood bank of KPK. Material & Methods: Blood from twenty healthy volunteer donors was taken and kept in CPDA-1 containing blood bags at Institute of Basic Medical Sciences (IBMS), Khyber Medical University (KMU). Hb-levels and Erythrocyte, Reticulocyte counts, Mechanical Fragility Index (MFI) and immunofluorescence staining for ankiyrin1 protein were performed on fresh blood samples. Samples for reticulocyte count was taken for 5 consecutive days while for the remaining parameters, blood was taken at 5 days interval till day 20th. The light and fluorescence micrographs were obtained accordingly and osmotic fragility tests were performed. Results: A significant mean reduction in erythrocyte counts and Hb-level was observed from day 0 to 20 (p=0.001), while MFI increased from day 0 to day 20 (12.88%±7.58 p=0.001). Reticulocyte count also decreased from day 0 up to day 5 (p=0.001). A weak association was observed between changes in MFI and erythrocyte morphology from day 0 to 20 (r=0.349), while the intensity and pattern of ankyrin1 protein expression appeared to change from day 10. Conclusion: Blood stored for a week has same properties as fresh blood, however, important structural alterations start to appear after the first week of storage and worsen with time. Therefore, to gain better transfusion results, blood stored for up to one week can safely be transfused.

Background: Red cell concentrate (RCC) cryopreservation allows for long-term storage of RCCs with rare phenotypes. Currently, tubing segments are not produced for these frozen units. Pre-transfusion compatibility testing therefore requires thawing and deglycerolization of the whole unit. A study was conducted to demonstrate the feasibility of using segments for compatibility testing, including circumstances where segments would require shipment to a reference laboratory. Study Design and Methods: RCCs produced using the red cell filtration method from citrate-phosphate-dextrose whole blood collections were glycerolized (40%) at day 21 post-collection and segments were generated prior to freezing. Room temperature (RT, 18°C-20°C) or water bath (WB, 37°C) thawing of segments was performed prior to storage at RT or at refrigerated temperatures (cold, 1°C -6°C) for 0, 24, 48, or 72 hours followed by deglycerolization and hemolysis testing. Additional segments were thawed and shipped in temperature-controlled containers at either RT or 1°C -10°C for antibody screening. Results: Hemolysis and RBC recovery results did not show significant differences over the storage period or between thawing and storage conditions. RBC recovery ranged from 46% to 64%. Hemoglobin (Hb) recovery ranged from 56% to 96%; for RT-thawed segments, recovery was significantly higher at 24 hours and lower at 72 hours for RT storage compared with cold storage. WB-thawed, cold-stored segments had higher Hb recoveries at 48 hours. Phenotype assessment was successful for all segments regardless of thawing method or shipping condition. Discussion: The shipment of thawed segments containing glycerolized red cells is feasible for the purpose of conducting pretransfusion phenotype evaluations or pretransfusion compatibility checks.

Transfusion of donor blood is sometimes unavoidable during the surgical episode. However, it is associated with adverse outcomes. Interventions that improve preoperative haemoglobin and reduce the need for donor transfusion a have positive impact upon outcome. The principle of reducing the need for transfusion is known as patient blood management (PBM) and incorporates three pillars (anaemia management, limitation of blood loss and use of alternatives to donor blood products, and restrictive transfusion triggers) when used in combination appear the most effective way to limit the impact of surgical blood loss. [ABSTRACT FROM AUTHOR]

The antioxidant function of the phospholipid hydroperoxide glutathione peroxidase (GPx4) is vital for the homeostasis of many cell types, from neoplastic cells to normal erythroid precursors. However, some functional proteins in erythroid precursors are lost during the development of red blood cells (RBCs); whether GPx4 is maintained as an active enzyme in mature RBCs has remained unclear. Our meta-analyses of existing RBC proteomics and metabolomics studies revealed the abundance of GPx4 to be correlated with lipid-anchored proteins. In addition, GPx4 anti-correlated with lyso-phospholipids and complement system proteins, further supporting the presence of active GPx4 in mature RBCs. To test the potential biological relevance of GPx4 in mature RBCs, we correlated the rate of hemolysis of human RBCs during storage with the abundance of GPx4 and other heritable RBC proteins. Of the molecules that anti-correlated with the rate of hemolysis of RBCs, proteins that mediate the cellular response to hydroperoxides, including GPx4, have the greatest enrichment. Western blotting further confirmed the presence of GPx4 antigenic protein in RBCs. Using an assay optimized to measure the activity of GPx4 in RBCs, we found GPx4 to be an active enzyme in mature RBCs, suggesting that GPx4 protects RBCs from hemolysis during blood bank storage.

Purpose of the study — to evaluate biomechanical regularities of deep deformation of native erythrocytes’ membranes during long-term (up to 32 days) storage of erythrocyte suspension.Materials and methods. The method for addressing the said problem was atomic-force spectroscopy. The measured value was hHz comprizing the depth to which the probe immersion process was described by interaction with a homogeneous medium. Empirical and theoretical dependence of the interaction force F (nN) on the probe immersion depth h (nm) — F (h) were obtained. Bar charts of relative frequency density of Young’s modulus E were built.Results. Modulus E changed from 9.3±3.2 kPa — for 3 days of storage, to 22.7±8.7 kPa — for 32 days. Coefficients of skewness were 0.52±0.04 (for day 3) and 0.82±0.09 (for day 32 d), hHz value remaining constant.Conclusion. Progressively as erythrocyte suspension was stored, erythrocyte membranes to the depth of 700 nm deflected homogeneously in spite of 2.4-fold increase of Young’s modulus.

The storage lesions and the irradiation of blood cellular components for medical procedures in blood banks are events that may induce nanochanges in the membrane of red blood cells (RBCs). Alterations, such as the formation of pores and vesicles, reduce flexibility and compromise the overall erythrocyte integrity. This review discusses the alterations on erythrocytic lipid membrane bilayer through their characterization by confocal scanning microscopy, Raman, scanning electron microscopy, and atomic force microscopy techniques. The interrelated experimental results may address and shed light on the correlation of biomechanical and biochemical transformations induced in the membrane and cytoskeleton of stored and gamma-irradiated RBC. To highlight the main advantages of combining these experimental techniques simultaneously or sequentially, we discuss how those outcomes observed at micro- and nanoscale cell levels are useful as biomarkers of cell aging and storage damage.

INTRODUCTION: Gamma Irradiation of blood products accentuates biochemical changes in the blood stored at 4°C. This study tried to compare the changes in potassium, sodium, glucose, lactate, and lactate dehydrogenase (LDH) levels in packed red blood cell (PRBC) units irradiated at various time points and then stored versus those stored for a particular period then irradiated. METHODOLOGY: One hundred and eighty units of RBCs were randomly assigned equally to be irradiated or not. Eighteen units each were irradiated by gamma irradiator using cobalt 60 (BI 2000) on day 1, 7, 14, 21, and 28 of their storage, respectively, in the irradiation group. All the units were assessed for their plasma levels of potassium, sodium, glucose, LDH, and lactate by clinical chemistry auto analyzer Beckman coulter AU680 weekly. The values were documented and analyzed by SPSS. RESULTS: Baseline values on day 1 for studied biochemical parameters were comparable between irradiated and nonirradiated groups. Mean values of potassium, lactate and LDH were higher in irradiated than nonirradiated PRBC bags. In contrast, Sodium and Glucose mean values were lower than baseline values. Maximum cumulative mean values were noted in day-21 irradiated bags when the parameters were measured on day-28 for potassium and lactate levels. This was followed by day 14 irradiated bags, followed by day 7 irradiated bags. CONCLUSION: The study indicates that irradiation of red cells later in their storage period had comparatively more detrimental changes in relation to potassium and lactate than irradiation in earlier days. Consideration of irradiation to be performed as close to the issue as possible to reduce a lesser number of days of storage postirradiation is to be explored. [ABSTRACT FROM AUTHOR]

The storage lesions and the irradiation of blood cellular components for medical procedures in blood banks are events that may induce nanochanges in the membrane of red blood cells (RBCs). Alterations, such as the formation of pores and vesicles, reduce flexibility and compromise the overall erythrocyte integrity. This review discusses the alterations on erythrocytic lipid membrane bilayer through their characterization by confocal scanning microscopy, Raman, scanning electron microscopy, and atomic force microscopy techniques. The interrelated experimental results may address and shed light on the correlation of biomechanical and biochemical transformations induced in the membrane and cytoskeleton of stored and gamma-irradiated RBC. To highlight the main advantages of combining these experimental techniques simultaneously or sequentially, we discuss how those outcomes observed at micro- and nanoscale cell levels are useful as biomarkers of cell aging and storage damage. [ABSTRACT FROM AUTHOR]

As part of the ZOOMICS project, we set out to investigate common and diverging metabolic traits in the blood metabolome across various species by taking advantage of recent developments in high-throughput metabolomics. Here we provide the first comparative metabolomics analysis of fresh and stored human (n = 21, 10 males, 11 females), olive baboon (n = 20), and rhesus macaque (n = 20) red blood cells at baseline and upon 42 days of storage under blood bank conditions. The results indicated similarities and differences across species, which ultimately resulted in a differential propensity to undergo morphological alterations and lyse as a function of the duration of refrigerated storage. Focusing on purine oxidation, carboxylic acid, fatty acid, and arginine metabolism further highlighted species-specific metabolic wiring. For example, through a combination of steady state measurements and 13C615N4-arginine tracing experiments, we report an increase in arginine catabolism into ornithine in humans, suggestive of species-specific arginase 1 activity and nitric oxide synthesis—an observation that may impact the translatability of cardiovascular disease studies carried out in non-human primates (NHPs). Finally, we correlated metabolic measurements to storage-induced morphological alterations via scanning electron microscopy and hemolysis, which were significantly lower in human red cells compared to both NHPs.

White particulate matter has been an enigmatic entity inside blood bags. Frequent encounter with this uncommon phenomenon is described world over as a sporadic report or small series. Over the years, as its incidence keeps increasing so is the need to understand this strange entity that may provide valuable insights into the fascinating world of blood physiology and storage. An attempt to comprehend this strange phenomenon has been made in this brief review based on the literature available on this subject.

Hemoglobin (Hb) inside and outside the red blood cells (RBCs) undergoes constant transformation to an oxidized form in a process known as autoxidation. The ferrous heme iron (Fe2+) of the prosthetic group is spontaneously transformed into an oxidized ferric (Fe3+) form, but under oxidative stress conditions a higher oxidation ferryl heme (Fe4+) is also formed. Although Fe3+ is a non-functional form of Hb, the Fe4+ is also extremely reactive towards other biological molecules due to its high redox potential. The RBC contains an effective reductive machinery that maintains Hb in the functional form with little oxidation during its life span. The redox transformation of Hb occurs to a lesser extent in young RBCs; it may, however, have detrimental effects on the integrity of these cells during ex vivo storage or when RBCs are subjected to pathogen reduction processes. In this review, Hb oxidation reactions (“oxidative lesion”) will be described, including details of how these reactions might impact the clinical use of stored or processed blood for therapeutic purposes.

Background/Aims: Although red blood cells (RBCs) transfusions can be lifesaving, they are not without risk. RBCs storage is associated with the abnormal metabolism of glutathione (GSH), which may increase the risk of the oxidative damage of RBCs after transfusion. The responsible mechanisms remain unknown. Methods: We determined the L-cysteine efflux and influx by evaluating the changes of free -SH concentrations in stored RBCs. The glutamate cysteine ligase (GCL) activities and protein content in stored RBCs was determined by fluorescence assay and western blotting. In addition, the glucose metabolism enzyme activity of RBCs was measured by spectrophotometric assay under in vitro incubation conditions. Results: We found that both L-cysteine transport and GCL activity significantly declined, thereby inducing the dysfunction of GSH synthesis during blood storage, which could be attenuated by ATP supplement and DTT treatment. In addition, the glycometabolic enzyme (G6PDH, HK, PK and LDH) activity significantly decreased after 6 weeks storage. Oxidant stress-induced dysfunction in glucose metabolism was the driving force for decreased GSH synthesis during storage. Conclusion: These experimental findings reflect an underlying molecular mechanism that oxidant stress induced glucose metabolism dysfunction contribute to decreased GSH synthesis in stored RBCs.