Using redox potential as a feasible marker for banked blood quality and the state of oxidative stress in stored red blood cells

BackgroundStored red blood cells (RBCs) may undergo oxidative stress over time, with functional changes affecting oxygen delivery. Central to these changes are oxidation‐reduction (redox) reactions and redox potential (RP) that must be maintained for cell function. RP imbalance can lead to oxidative stress that may contribute to storage lesions. This study''s purpose was to identify changes in RP over time in banked RBCs, and among RBCs of similar age.MethodsMultiple random RBC segments from RBC units were tested (n = 32), ranging in age from 5 to 40 days, at 5‐day intervals. RP was recorded by measuring open circuit potential of RBCs using nanoporous gold electrodes with Ag/AgCl reference. RP measures were also performed on peripheral venous blood from 10 healthy volunteers. RP measures were compared between RBC groups, and with volunteer blood.ResultsStored RBCs show time‐dependent RP increases. There were significant differences in Day 5 RP compared to all other groups (p ≤ 0.005), Day 10–15 vs. ages ≥ Day 20 (p ≤ 0.025), Day 20–25 vs. Day 40 (p = 0.039), and all groups compared to healthy volunteers. RP became more positive over time suggesting ongoing oxidation as RBCs age; however, storage time alone was not predictive of RP measured in a particular unit/segment.ConclusionsThere are significant differences in RP between freshly stored RBCs and all others, with RP becoming more positive over time. However, storage time alone does not predict RP, indicating RP screening may be an important measure of RBC oxidative stress and serve as an RBC quality marker.     

Autologous red cells derived from cord blood: collection, preparation, storage and quality controls with optimal additive storage medium (Sag-mannitol)

To investigate whether packed red cells (PRCs) prepared from autologous cord blood-packed red cells (AC-PRCs) could be used as an alternative for homologous-packed red cells (H-PRCs), we developed a system to collect and prepare AC-PRCs and determined standard storage parameters during 35 days of storage in extended storage medium (Sag-mannitol). We collected and fractionated cord blood from 390 newborns. The amount and quality of the AC-PRCs were analysed. The bacterial contamination rate was 1.84%. Twelve AC-PRCs were stored for 35 days, and standard laboratory parameters were measured at day 1 and day 35. The initial laboratory parameters of the AC-PRCs were similar to the parameters of the H-PRCs. After 35 days, the AC-PRCs displayed an increased haemolysis rate compared to H-PRCs (1.1 versus 0.2%) and also a significant decreased adenosine triphosphate value (1.2 versus 2.3 micromol L(-1)). Haemoglobin, haematocrit and pH were comparable in both groups. AC-PRCs meet the quality criteria for H-PRCs after 35 days. Utilizing a closed collection system for cord blood and an extended storage medium will increase safety and quality and facilitate the routine transfusion of autologous red cells derived from cord blood.     

The preservative-exchange method using a sextuple-bag system for a 10-week storage period of red blood cells

Summary. Prolonged storage of red blood cells in a liquid state was achieved by replacing the preservative using a sextuple-bag system. The bag system consists of one primary bag containing citrate-phosphate-dextrose (CPD) solution, three satellite bags containing saline-adenine-glucose-phosphate-maltose (SAGP-maltose) solution, and two empty satellite bags to remove plasma and buffy coat. Preservative can be exchanged three times in this closed system. The system is able to supply nutrients, such as glucose, and to remove harmful metabolites, such as lactic acid, by exchanging the preservative during storage. As a result, red cells stored by this method showed much higher levels of total adenylate and morphological score after the second preservative exchange, when compared with red cells stored by the conventional method ( P < 0·01). Judging from these two in-vitro parameters, red cells may tolerate storage for at least 10 weeks in a liquid state. This method might be useful for 'predeposit autologous transfusion', as it is more convenient and more cost effective than the freeze-preservation method.     

The experience of flow cytometry for specific antibody against cisplatin-treated red blood cells: A case report

Background

Cisplatin-associated hemolysis is a rare but important adverse effect. Nonimmunological protein adsorption (NIPA) due to erythrocyte membrane modification has been reported as the leading cause of cisplatin-associated hemolysis. However, limited data exist on cisplatin-associated immunological hemolysis because of a lack of an established diagnostic method. Here, we used flow cytometry (FCM) to diagnose a patient with cisplatin-associated immunological hemolysis.

Study Design and Methods

A 55-year-old woman with uterocervical cancer was treated with weekly cisplatin monotherapy (40 mg/m²). She had no previous transfusion and medication history, nor any significant family history. On the 26th day after cisplatin administration, severe hemolysis was noted. Her red blood cells (RBCs) and sera were evaluated by direct antiglobulin test (DAT) and indirect antiglobulin test (IAT), respectively. To explore immunological reactions for cisplatin-treated RBCs, we attempted FCM using cisplatin-treated and -untreated RBCs. After incubating conditioned RBCs with the patient's serum or healthy donor serum, we evaluated their fluorescent intensity by fluorescein isothiocyanate (FITC)-conjugated anti-human immunoglobulin (Ig) G antibodies.

Results

The patient's DAT was positive, and an IAT using her plasma was positive for cisplatin-treated RBCs. FCM using cisplatin-treated RBCs revealed that the patient's serum had higher FITC intensity than the donor's serum, indicating the existence of cisplatin-treated RBC-specific IgGs in patient's serum.

Conclusion

Here, we report a rare case of a patient with hemolysis diagnosed using FCM to identify specific antibodies against cisplatin-treated RBCs. NIPA and immunological mechanisms may contribute to hemolysis onset during cisplatin treatment.     

Prolonged maintenance of 2,3-diphosphoglycerate acid and adenosine triphosphate in red blood cells during storage

BACKGROUND: Current additive solutions (ASs) for red cells (RBCs) do not maintain a constant level of critical metabolites such as adenosine triphosphate (ATP) and 2,3-diphosphoglycerate acid (2,3-DPG) during cold storage. From the literature it is known that the intracellular pH is an important determinant of RBC metabolism. Therefore, a new, alkaline, AS was developed with the aim to allow cold storage of RBCs with stable product characteristics. STUDY DESIGN AND METHODS: Whole blood-derived RBCs (leukoreduced) were resuspended in experimental medium phosphate-adenine-guanosine-glucose-gluconate-mannitol (PAGGG-M; pH 8.2) with and without washing in the same medium. During cold storage several in vitro variables, such as intracellular pH, 2,3-DPG, ATP, and hemolysis, were analyzed. RESULTS: During cold storage, RBCs resuspended in PAGGG-M showed a constant ATP level (approx. 6 mumol/g Hb) and a very limited hemolysis (<0.2%). The 2,3-DPG content showed an increase until Day 21 (150% of initial level), followed by a slow decrease, with at Day 35 still 100 percent of the initial level. RBCs washed in PAGGG-M even showed a continuous increase of 2,3-DPG during 35 days, with a maximum level of 200 percent of the initial value. The effect of PAGGG-M appears to be related to long-lasting effects of the initial intracellular pH shortly after production. CONCLUSION: Resuspension of RBCs in our alkaline medium PAGGG-M resulted in a RBC unit of high quality during storage for up to at least 35 days, with 2,3-DPG levels of higher than 10 mumol per g Hb, hemolysis of less than 0.2 percent, and ATP levels of higher than 5 mumol per g Hb.     

福州地区小儿小规格红细胞临床使用情况分析

目的统计分析福州地区小儿小规格红细胞临床使用情况,更好地为临床小儿输血提供服务。方法回顾性分析福州地区2011—2018年0.5 U红细胞的临床用血情况,采用统计学方法分析"健康中国"战略、"全面二孩"政策颁布前(2011—2014年)后(2015—2018年)的临床用血量是否存在差异,并对使用量最大的3所医院进行分析。结果福州地区2015—2018年0.5 U红细胞供应量较2011—2014年增长54.70%(P<0.05),其中市区医院增长54.64%,郊县医院增长87.50%。市区医院0.5 U红细胞供应量占0.5 U红细胞总供应量99.8%。供应量最多的3所医院占0.5 U红细胞总供应量89.60%,这3所医院2015—2018年的供应量较2011—2014年分别增长41.85%、49.44%、128.13%(P<0.05)。结论国家颁布"健康中国"、"全面二孩"政策后福州地区0.5 U红细胞需求量较前增长。加强血液管理信息系统建设、与医院增强输血相关信息共享、尝试开展其他小规格的红细胞分装制备是今后小儿血液供应工作可探寻的方向。     

创伤患者输血后发生细菌感染的危险因素探讨

目的探讨创伤患者在输注红细胞或血小板后发生细菌感染的危险因素。方法对2007年1月至2012年1月在我院输注红细胞或血小板后发生细菌感染的56例创伤患者(感染组)与输血后未发生感染的140例创伤患者(未感染组),采用多元回归方法对感染危险因素进行分析。结果创伤患者输血后感染与输注保存14天以上的红细胞密切相关[OR1.038(95%CI:1.01～1.07),P=0.036],而与输注红细胞的数量或血小板的数量无关。结论输注保存时间超过14天的红细胞是创伤患者输血后感染的一个重要危险因素。     

Use of red cells preserved in extended storage media for exchange transfusion in anti-k haemolytic disease of the newborn

Anti-k is a Kell-related antibody. There is little correlation between the maternal antibody titre and the severity of haemolytic disease of the foetus and newborn, and anaemia is usually associated with low bilirubin levels. Severe erythroblastosis has been reported with a low titre anti-k (IAT 8-16). We report a case of severe haemolytic disease of the newborn (HDN) due to anti-k. HDN was associated with a normal bilirubin level and reticulocytopenia. The foetus was monitored by ultrasound, and delivery by elective caesarean section (CS) was planned. The mother was admitted 1 week before the expected date of delivery, and the infant was delivered by urgent CS. The infant required exchange transfusion. As suitable plasma-reduced (k antigen(-)) red cell units were not readily available, k- SAGM red cell units (preserved in extended storage media: SAGM sodium chloride, adenine, glucose and mannitol) were provided. The post-transfusion Hb remained stable, and the infant did not require further transfusion support. Our findings (reticulocytopenia and normal bilirubin levels) support the hypothesis that the pathogenesis of anaemia and haemolysis in anti-k HDN may be similar to that in anti-K (suppression of erythropoesis and immune destruction of K+ erythroid progenitor cells by macrophages in the foetal liver). The ideal product for exchange transfusion is plasma-reduced RBC, less than 5-days old. We provided a 4-day-old SAGM red cell unit for exchange transfusion in a term infant, and this was uneventful. Caution should be taken, however, and renal function and electrolyte levels should be monitored closely. More information is required regarding the safety of SAGM units for exchange transfusion.     

去白细胞悬浮红细胞制剂储存过程中活性氧族及丙二醛含量变化

目的 探讨去白细胞悬浮红细胞制剂储存过程中活性氧族(ROS)含量及丙二醛(MDA)含量的变化.方法 从2013年5～6月于河北省血液中心制备的去白细胞悬浮红细胞制剂中,采用简单随机抽样方法抽取6份去白细胞悬浮红细胞制剂标本作为研究对象.于储存后第0,7,14,28,42天,采用荧光探针2',7'-二氧荧光黄双乙酸盐(DCFH-DA)孵育的方法观察去白细胞悬浮红细胞荧光强度,并对去白细胞悬浮红细胞制剂内ROS含量与MDA含量进行检测.对储存后去白细胞悬浮红细胞制剂内ROS含量、MDA含量分别与储存时间进行相关性分析;并对储存后去白细胞悬浮红细胞制剂内ROS含量与MDA含量进行相关性分析.结果 ①随着储存时间的延长,经荧光探针DCFH-DA孵育的去白细胞悬浮红细胞的荧光强度显著增强;②储存后第0,7,14,28,42天去白细胞悬浮红细胞制剂内ROS含量呈显著增高趋势,第42天去白细胞悬浮红细胞内ROS含量显著高于第0,7,14,28天ROS含量,并且差异均有统计学意义(P＜0.01,0.05,0.05,0.05);且去白细胞悬浮红细胞制剂内ROS含量与储存时间呈正相关关系(r=0.898,P＜0.01);③储存后第0,7,14,28,42天去白细胞悬浮红细胞内MDA含量呈显著增高趋势,第42天去白细胞悬浮红细胞内MDA含量显著高于第0,7,14,28天MDA含量,并且差异均有统计学意义(P＜0.01,0.01,0.01,0.05);且去白细胞悬浮红细胞内MDA含量与储存时间呈正相关关系(r=0.930,P＜0.01);④储存后第0,7,14,28,42天,去白细胞悬浮红细胞制剂内ROS含量均分别与对应时间点MDA含量呈正相关关系(r=0.877,0.872,0.823,0.786,0.907;P＜0.05).去白细胞悬浮红细胞制剂储存后第0,7,14,28,42天,各时间点ROS含量均值与MDA含量均值亦呈正相关性关系(r=0.981,P＜0.01).结论 去白细胞悬浮红细胞制剂储存过程中ROS含量与MDA含量均显著增高,红细胞内氧化应激反应增强,其为导致去白细胞悬浮红细胞制剂储存损伤的重要原因.     

川芎嗪对回收红细胞和机体凝血功能影响的研究

目的观察川芎嗪对回收自体血中红细胞及回输血后机体凝血功能的影响，评价提高红细胞回收率的价值。方法40例择期脊柱手术患者，随机分成两组，试验组于收集血液前静脉滴注川芎嗪，并在肝素盐水和洗涤盐水内加入川芎嗪。检测回输血液样本的红细胞（RBC）、血红蛋白（Hb）、红细胞压积（HCT），计算红细胞回收率；测定术前、回输血后即刻、术后24h的血常规、凝血酶原时间（PT）、部分凝血活酶时间（APTT）、纤维蛋白原（FiB）；描记术前、回输血后即刻血栓弹性描记图（TEG）。对照组不予静脉滴注川芎嗪，肝素盐水和洗涤盐水内不加川芎嗪。结果试验组红细胞回收率高于对照组（75．3％±8．3％vs66．5％±5．3％，P〈0．01）；与术前比较，试验组回输血后Hb、HCT、PLT计数明显降低（P〈0．01），PT、APTT、反应时间（R）显著延长（P〈0．01），FiB和最大宽幅（MA）降低（P〈0．01），但两组变化趋势一致。结论川芎嗪能提高红细胞回收率，并且未对凝血功能产生明显的影响。     

Clinical implications of using non-invasive haemoglobin monitoring for red blood cell transfusion decision in hip arthroplasty

IntroductionThe decision to transfuse red blood cells requires accurate haemoglobin concentration values. In this study, we evaluated if continuous non-invasive haemoglobin (SpHb) measurement could substitute laboratory determined haemoglobin (LabHb) in patients undergoing elective hip replacement. As secondary objective, we analyzed the trend of the difference between techniques.Materials/methodsLabHb measurements were done using an automated analyser and SpHb measurements were acquired using Radical-7®. In randomly selected patients undergoing hip replacement, whenever blood was collected for LabHb, concomitant SpHb was recorded. Correlation, bias and accuracy of SpHb were calculated in comparison with LabHb.Results108 paired measurements were obtained from 43 patients. The Pearson R of the correlation between SpHb and LabHb was 0.7 (p < 0.001). Bland-Altman test revealed a bias of 1 ± 1.4 g dL⁻¹, meaning Lab Hb was recurrently higher than SpHb. Limits of agreement were [−1.7; 3.8]. Considering RBC transfusion threshold of 8 g dL⁻¹, we found that in two situations transfusion decision would differ based on the measurement considered. Trending ability of SpHb study showed a significant difference between preoperative and postoperative LabHb-SpHb.DiscussionThere was a good correlation between SpHb and LabHb, while bias and limits of agreement were higher than those in literature. There was a limited trending ability of SpHb during the perioperative period. Despite this, using SpHb instead of LabHb for decision making regarding transfusion would only change the decision in 1.9 % of our cases. Our findings suggest that this device could be used as a reference but cannot replace venous puncture as gold standard.     

Whole blood versus red cells and plasma for exchange transfusion in ABO haemolytic disease

Records of 381 neonates who underwent exchange transfusion (ET) due to ABO haemolytic disease at the Division of Neonatology of Hacettepe University, Ankara, Turkey, between January 1977 and December 2003 were reviewed. Records were kept for the type of blood used in ET, the number of ETs for each infant, adverse event attributable to ET and bilirubin levels before, and 4 and 8 h after each ET. Of 381 infants, 300 were transfused with whole blood, whereas 81 infants were transfused with O red cells suspended in A or B plasma. The re-exchange rate was higher in the whole blood group, compared with the erythrocyte and plasma group. Use of erythrocyte and plasma provided 30% reduction in the number of ETs per patient. Eight hours after the first ET, mean bilirubin levels were 84% of the pre-exchange values in the whole blood group and 73% of the pre-exchange values in the erythrocyte and plasma group (P = 0.001). As the use of O group red cells re-suspended in AB plasma decreased the re-exchange risk compared with O group whole blood, we suggest the use of O red cells re-suspended in AB plasma for the ET in cases of ABO haemolytic disease.     

Multifaceted role of glycosylation in transfusion medicine,platelets, and red blood cells

Glycosylation is highly prevalent, and also one of the most complex and varied posttranslational modifications. This large glycan diversity results in a wide range of biological functions. Functional diversity includes protein degradation, protein clearance, cell trafficking, cell signaling, host‐pathogen interactions, and immune defense, including both innate and acquired immunity. Glycan‐based ABO(H) antigens are critical in providing compatible products in the setting of transfusion and organ transplantation. However, evidence also suggests that ABO expression may influence cardiovascular disease, thrombosis, and hemostasis disorders, including alterations in platelet function and von Willebrand factor blood levels. Glycans also regulate immune and hemostasis function beyond ABO(H) antigens. Mutations in glycogenes (PIGA, COSMC) lead to serious blood disorders, including Tn syndrome associated with hyperagglutination, hemolysis, and thrombocytopenia. Alterations in genes responsible for sialic acids (Sia) synthesis (GNE) and UDP‐galactose (GALE) and lactosamine (LacNAc) (B4GALT1) profoundly affect circulating platelet counts. Desialylation (removal of Sia) is affected by human and pathogenic neuraminidases. This review addresses the role of glycans in transfusion medicine, hemostasis and thrombosis, and red blood cell and platelet survival.