A pilot study of the possibility and the feasibility of haemoglobin dosing with red blood cells transfusion

Background and Objectives Red blood cell concentrates (RBCs) are the major blood component transfused. Although the haemoglobin content is variable, the transfusion dose is prescribed as units of red cell concentrates. Thus, by chance, large volume patients may receive a low haemoglobin dose and low volume patients may be transfused with haemoglobin‐rich RBCs. The aim of this study was to evaluate whether the haemoglobin increment (grams per litre) in the patient can be predicted from the haemoglobin dose (in grams) transfused, with and without correction for estimated blood volume. If this is true, it may be possible to achieve the predicted transfusion outcome by selecting RBCs for each patient. Materials and Methods Haemodynamically stable patients scheduled for day treatment with transfusion of RBCs were recorded. A total of 52 transfusions episodes, 27 for women and 25 for men, were recorded. Blood volumes were estimated, haemoglobin content in the RBCs was measured before transfusion, and pre‐ and post‐transfusion haemoglobin concentrations were obtained. Results The haemoglobin content of the RBCs prepared for transfusion showed a wide range, varying from 38·7 g/unit to 69·0 g/unit. There were statistically significant correlations between haemoglobin concentration in the RBCs and haemoglobin increment in patients. Conclusion Post‐transfusion increment in circulating haemoglobin can be predicted from the haemoglobin content of transfused cells, but knowledge of the patient’s blood volume improves the accuracy of prediction. It may be feasible to select the high haemoglobin content RBC for patients with largest blood volume and vice versa.     

Immunochemical estimation of haemoglobin types in red blood cells by FACS analysis

Summary. A fixation and permeabilization procedure using formaldehyde and acetone has been developed which allows immunostaining of intracellular haemoglobin for fluorescence activated cell sorter (FACS) analysis of erythrocytes. The treatment preserves antigenicity and light-scattering properties. Validation of the method was given by the correlation of F cell number in adults determined by FACS analysis with that assessed by microscopic examination of cell smears, and by the direct relationship between β chain synthesis and intensity of β chain/Hb A immunofluorescence within fetal erythrocyte samples known to vary in their β chain/Hb A content. The procedure is rapid, non-subjective and sensitive, and makes analysis of haemoglobin content, type and distribution amongst red cell populations possible.     

Spurious counts and spurious results on haematology analysers: a review. Part II: white blood cells, red blood cells, haemoglobin, red cell indices and reticulocytes

Haematology analysers provide quick and accurate results in most situations. However, spurious results, related either to platelets (part I of this report) or to other parameters from the cell blood count (CBC) may be observed in several instances. Spuriously low white blood cell (WBC) counts may be observed because of agglutination in the presence of ethylenediamine tetra-acetic acid (EDTA). Cryoglobulins, lipids, insufficiently lysed red blood cells (RBC), erythroblasts and platelet aggregates are common situations increasing WBC counts. In most of these instances flagging and/or an abnormal WBC differential scattergram will alert the operator. Several situations lead to abnormal haemoglobin measurement or to abnormal RBC count, including lipids, agglutinins, cryoglobulins and elevated WBC counts. Mean (red) cell volume (MCV) may be also subject to spurious determination, because of agglutinins, excess of glucose or salts and technological considerations. In turn, abnormality related to one measured parameter will lead to abnormal calculated RBC indices: mean cell haemoglobin content (MCHC) is certainly the most important RBC indices to consider, as it is as important as flags generated by the haematology analysers (HA) in alerting the user to a spurious result. In many circumstances, several of the measured parameters from CBC may be altered, and the discovery of a spurious change on one parameter frequently means that the validity of other parameters should be considered. Sensitive flags now allow the identification of several spurious counts, but only the most sophisticated HA have optimal flagging and more simple HA, especially those without a WBC differential scattergram, do not possess the same sensitivity for detecting anomalous results. Reticulocytes are integrated now into the CBC in many HA, and several situations may lead to abnormal counts.     

Vacuoles in red cells from splenectomized subjects originate during cell life: Association with glycosylated haemoglobin?

Whole blood from splenectomized subjects (n = 8) contained a substantial percentage of vacuolated ('pitted') red cells (34.7 +/- 8.3%), while blood from controls revealed none. The percentage of haemoglobin A1 (HbA1) had increased significantly compared with controls (p less than 0.01). Fractionation on cell density revealed that the number of 'pitted' cells and the HbA1 percentage were associated with increased cell density. Fractionation on cell volume showed that 'pitted' cells are equally distributed in fractions with varying mean cell volume (MCV) and that decreasing MCV was associated with a linear rise of HbA1. It appeared that, shortly after splenectomy (n = 4), 'pits' develop early in cell life and that also older cells, after previous splenic contact, are capable of pit formation. A positive correlation found between the number of 'pitted' cells and the percentage of HbA1 might point to an impaired release of HbA1, manifest in the presence of 'pits'. The increased percentage of HbA1 in whole blood from splenectomized subjects may thus be explained.     

Preferential localisation of red cell vacuoles to pathologically shaped human red blood cells

The percentage of pitted erythrocytes and Howell-Jolly bodies in peripheral blood samples of 51 individuals following posttraumatic splenectomy and 20 patients splenectomized because of various haematological diseases differed significantly from each other (p less than 0.001) and from that of healthy controls (p less than 0.001). The percentage of pitted erythrocytes was significantly higher in pathologically shaped red blood cells (RBCs) (acanthocytes, schizocytes, elliptocytes) than in normal discoid shaped RBCs (p less than 0.001). As the number of pits per RBC showed great individual variations, a scoring system for the evaluation of pitted RBCs is proposed.     

Sickle cell disease: relation between procoagulant activity of red blood cells from different phenotypes and in vivo blood coagulation activation

In the present study we examined if, among other mechanisms, the abnormal exposure of phosphatidylserine at the surface of sickle red blood cells (RBCs) contributes to the hypercoagulability which characterizes homozygous sickle cell disease (SCD). The question was addressed by comparison of the procoagulant properties of RBCs from subjects with various phenotypes (SS, SC and AS) that differ in clinical presentation. As previously reported, SS-RBCs accelerated the prothrombin activation by factor Xa, by decreasing the K _m of the reaction compared to normal RBCs. SC-RBCs and AS-RBCs also promoted prothrombin activation although their procoagulant properties were milder compared to SS-RBCs. A significant increase of the thrombin–antithrombin complexes was observed in SS subjects. Prothrombin fragment 1+2 (F1+2) was elevated in half of the SS subjects, but the difference with controls did not reach significance. Elevated levels of thrombin–antithrombin complexes were observed in a number of SC (4/11) and AS (3/12) subjects, but the difference with controls was not significant. A significant correlation was observed between the plasma levels of thrombin–antithrombin complexes in the subjects with SS, AS and AA phenotypes, and the procoagulant properties of RBCs. Our results strongly suggest that the procoagulant properties which characterize SS-RBCs also affect SC-RBCs and AS-RBCs, and that exposure of phosphatidylserine by RBCs contributes to the hypercoagulable state observed in SCD.     

Evaluation of the HemoCue plasma haemoglobin analyser for assessing haemolysis in red cell concentrates during storage

BACKGROUND AND OBJECTIVES: To evaluate the HemoCue plasma haemoglobin (Hb) analyser for determining supernatant Hb in red cell concentrates (RCC) during storage and compare this to a spectrophotometric method. MATERIALS AND METHODS: Samples from RCC (n = 20) at days 0, 7, 21, 35 and 42 of storage were tested by both methods for supernatant Hb. RESULTS: Results are given as median with range. There was a significant correlation (r = 0.97, P < 0.0001), but statistically significant difference between the two methods (0.72 (0.36-2.77) spectrophotometric vs. 0.70 (0.3-2.6) g/l HemoCue). However, the mean bias was negligible at 0.06 g/l. The HemoCue method gave a interassay coefficient of variation of 5.6% at 1.2 g/l and was linear to at least 20 g/l. CONCLUSION: The HemoCue plasma haemoglobin analyser is a simple, rapid, reliable and accurate method for the determination of supernatant Hb in RCC at the end of storage and compares well to established methods for supernatant Hb measurement.     

Effects of helicopter transport on red blood cell components

Background

There are no reported studies on whether a helicopter flight affects the quality and shelf-life of red blood cells stored in mannitol-adenine-phosphate.

Materials and methods

Seven days after donation, five aliquots of red blood cells from five donors were packed into an SS-BOX-110 container which can maintain the temperature inside the container between 2 °C and 6 °C with two frozen coolants. The temperature of an included dummy blood bag was monitored. After the box had been transported in a helicopter for 4 hours, the red blood cells were stored again and their quality evaluated at day 7 (just after the flight), 14, 21 and 42 after donation. Red blood cell quality was evaluated by measuring adenosine triphosphate, 2,3-diphosphoglycerate, and supernatant potassium, as well as haematocrit, intracellular pH, glucose, supernatant haemoglobin, and haemolysis rate at the various time points.

Results

During the experiment the recorded temperature remained between 2 and 6 °C. All data from the red blood cells that had undergone helicopter transportation were the same as those from a control group of red blood cell samples 7 (just after the flight), 14, 21, and 42 days after the donation. Only supernatant Hb and haemolysis rate 42 days after the donation were slightly increased in the helicopter-transported group of red blood cell samples. All other parameters at 42 days after donation were the same in the two groups of red blood cells.

Discussion

These results suggest that red blood cells stored in mannitol-adenine-phosphate are not significantly affected by helicopter transportation. The differences in haemolysis by the end of storage were small and probably not of clinical significance.     

手工分少白细胞红细胞和悬浮红细胞输注疗效观察

目的:了解并比较手工分少白细胞红细胞和悬浮红细胞的治疗效果。方法:将2 582例患者分为输注悬浮红细胞组(1 450例)和输注手工分少白细胞红细胞组(1 132例),输注前测定Hb值,复查患者及献血者ABO和RH(D)血型并作凝聚胺法交叉配血试验,输血后给每例患者进行血液学分析,检查Hb值,然后再进行输血不良反应发生率统计。结果:输注手工分少白细胞红细胞效果好于普通悬浮红细胞,并且输注手工分少白细胞红细胞的输血不良反应发生率明显减少。结论:输注手工分少白细胞红细胞可以使输血后非溶血性输血反应发生率降低。     

Net haemoglobin increase from reinfusion of refrigerated vs. frozen red blood cells after autologous blood transfusions

BACKGROUND AND OBJECTIVES Two main blood storage procedures can be used for storing red blood cells: refrigeration and freezing. Nevertheless, the efficiency of these procedures measured as the increase in haemoglobin after reinfusion compared with baseline has never been examined. The main objective was to examine which storage procedure yielded the largest increase in circulating haemoglobin after reinfusion compared to baseline. MATERIALS AND METHODS Equal volumes of blood from 15 men were withdrawn and stored either frozen or refrigerated as packed red blood cells. Serial measures of circulating haemoglobin by carbon monoxide rebreathing provided an opportunity to monitor recovery from anaemia, as well as the net increase in circulating haemoglobin after transfusion. RESULTS The post-thaw yield of haemoglobin in the bags was 72% after refrigerated storage compared with only 52% after freezing. Nevertheless, frozen storage allowed haemoglobin to fully recover before reinfusion, while the haemoglobin was 10% lower in the refrigerated group compared with baseline. After reinfusion, the haemoglobin levels were 11·5% higher than the baseline values in the group reinfused with frozen blood, while for the refrigerated group, haemoglobin levels were only 5·2% higher than baseline. CONCLUSION The relatively larger recovery from anaemia in the frozen group during storage more than compensated for the larger loss of haemoglobin during freezing and resulted in a larger net gain in haemoglobin. Based on the average 23 g per week recovery of haemoglobin, extending refrigerated storage to 7-8 weeks may yield sufficient time for patients to fully replenish harvested haemoglobin from three bags of blood without reliance on frozen storage of RBC.     

冰冻解冻去甘油红细胞研究进展

<正>冰冻解冻去甘油红细胞主要用于稀有血型血液及自体血液的长期保存,可减少输血反应,避免酸中毒、高钾血症及输血传播疾病,可杜绝因缺血引发的死亡事件,冰冻解冻去甘油红细胞制剂的作用是其他制剂所不能替代的,本文介绍冰冻解冻去甘油红细胞的研究进展。1冰冻解冻去甘油红细胞的应用范围目前冰冻解冻去甘油红细胞主要用途是:稀有血型患者的临床输血,Rh(D)阴性患者输血,其他稀有血型患者输血;自身输血,ABO亚型、孟买血     

The mean red cell volume in diabetes mellitus

Summary The Coulter mean red cell volume was found to be significantly elevated in a sample of 100 diabetic patients compared with 200 normal subjects (p < 0.01). There was no correlation between the mean red cell volume level and the type of diabetes, its method of treatment or degree of control as measured by random blood glucose and glycosylated haemoglobin levels.     

单个胎儿有核红细胞的DNA分析

目的：获取孕妇外周血中的极少量胎儿有核红细胞（ＮＲＢＣ）,并进行单个ＮＲＢＣ的基因分析,探讨其在产前基因诊断中的价值。方法：将孕妇外周血进行不连续密度梯度离心,将分离后的细胞进行制片,光学显微镜下识别ＮＲＢＣ,然后用显微操作仪获取单个ＮＲＢＣ进行引物延伸预扩增（ＰＥＰ）及Ｙ染色体特异性ＤＹＺ１基因的聚合酶链反应（ＰＣＲ）,以确定其来源于胎儿。结果：单个ＮＲＢＣ的ＰＥＰ－ＰＣＲ法检测结果,６０％检出     

Some characteristics of human red blood cells separated according to their size: a comparison with density-fractionated red blood cells

During their in vivo ageing, red blood cells (RBC) increase in density and become smaller. Age-defined RBC subpopulations are usually collected by centrifugation. A fractionation according to RBC volume has been proposed as an improved alternative to such age separation. Because a few data reported in the literature indicate some discrepancies between the two methods, blood samples were separated either by centrifugation or by counterflow centrifugation, and some characteristics of the RBC thus fractionated were studied. The enzyme activities decrease either when the density rises or when the volume (MCV) decreases. However, the comparison of other RBC characteristics strongly suggests that these two procedures do not lead to the collection of the same RBC subpopulations: for instance, the hemoglobin content increases when the MCV rises, whereas it remains constant whatever the RBC density is. With radiolabelled cells, it is shown 1) that the most dense RBC are recovered in all the size-separated RBC subpopulations, even though they tend to concentrate in the fractions with the largest MCV, and 2) that the smallest RBC are almost fairly distributed in all the RBC subpopulations, whatever their density, whereas the largest RBC are mainly, but not exclusively, present in the high-density fractions. Thus, fractionation according to size does not match separation according to density. Taken together with results from in vivo experiments carried out in mice and with the fact that reticulocytes are present in all the size-separated fractions, these data suggest that counterflow centrifugation may be a very questionable procedure to achieve a RBC fractionation according to age and therefore that RBC volume might not be a reliable criterion of RBC age.     

Red blood cell substitutes

Soluble polymerized haemoglobin (polyhaemoglobin) is now in a phase III clinical trials. Patients have received up to 20 units (10 litres) in trauma surgery and other surgery. Polyhaemoglobin can be stored for more than 1 year. Haemoglobin solutions have no blood group antigen and can be used as a 'universal donor' oxygen carrier. They can also be sterilized. With a circulation half-life of 24 hours they are undergoing trials for peri-operative use. For conditions with potential for ischaemia-reperfusion injuries, a new polyhaemoglobin-superoxide dismutase-catalase, which can reduce oxygen radicals, is being developed. Recombinant human haemoglobin has been tested in clinical trials, and a new type of recombinant human haemoglobin that has low affinity for nitric oxide is being developed for clinical trials. To increase the circulation time, artificial red blood cells have been prepared with a bilayer lipid membrane (haemoglobin liposomes) or with a biodegradable polymer membrane-like polylactide (haemoglobin nanocapsules). Synthetic chemicals such as perfluorochemicals are also being developed and tested in clinical trials as red blood cell substitutes.