Red Cell Recovery Research Articles

Captopril modulates hypoxia-inducible factors and erythropoietin responses in a murine model of total body irradiation

Our laboratory reported that the angiotensin converting enzyme inhibitor captopril improves erythroid recovery from total body irradiation (TBI) in mice when administered after irradiation. However, captopril administered before TBI attenuates erythroid recovery. Here we investigate captopril and radiation regulation of erythropoietin (EPO) and thrombopoietin (TPO), key effectors of erythroid progenitor proliferation and differentiation. C57BL/6 mice, nonirradiated or exposed to 7.5 Gy TBI ((60)Co, 0.6 Gy/min) were untreated or administered captopril. Plasma EPO and TPO levels were measured by enzyme-linked immunosorbent assay. Gene expression of EPO was determined by quantitative reverse transcription polymerase chain reaction. The hypoxia-inducible factors (HIF)-1α and -2α were measured by immunoblotting. In nonirradiated mice, continuous captopril administration in the water transiently reduced reticulocytes and red blood cells after 7 and 10 days, respectively. EPO plasma levels and gene expression were reduced below detectable limits after 2 days of captopril treatment, but recovered within 7 days. HIF-1α and HIF-2α were activated preceding reticulocyte and red blood cell recovery. TBI, which ablates early and late-stage erythroid progenitors, activated both HIFs and increased EPO and TPO. Captopril treatment postirradiation suppressed radiation-induced HIF activation and EPO expression. In contrast, captopril administration for 7 days before TBI resulted in earlier EPO induction and activation. Captopril treatment lowered TPO levels in nonirradiated mice, but had minimal effects on radiation-induced TPO. In nonirradiated mice, captopril biphasically regulates EPO via HIF activation. TBI ablates erythroid progenitors, resulting in hypoxia, HIF activation, and increased EPO expression that are modulated by captopril treatment. These data suggest that short-term suppression of radiation-induced EPO immediately after TBI is favorable for erythroid recovery.

Guidelines on the use of irradiated blood components prepared by the British Committee for Standards in Haematology blood transfusion task force

Search terms included: Transfusion-associated graft-versus-host disease, Transfusion-associated graft-versus-host, TA-GvHD. The last guideline covering this topic was published in 1996 (British Committee for Standards in Haematology (BCSH) Blood Transfusion Task Force, 1996. The writing group produced the new draft guideline, which was subsequently revised by consensus by members of the Haemato-oncology and Blood Transfusion Task Forces of the BCSH. The guideline was then reviewed by a sounding board of approximately 100 UK haematologists, the BCSH and the committee of the British Society for Haematology and amended, again by consensus. Criteria used to quote levels and grades of evidence are according to the GRADE system (Guyatt et al, 2006). Strong recommendations (grade 1, ‘recommended’) are made when there is confidence that the benefits either do or do not outweigh the harm and burden and costs of treatment. Where the magnitude of benefit or not is less certain a weaker grade 2 recommendation (‘suggested’) is made. Grade 1 recommendations can be applied uniformly to most patients whereas grade 2 recommendations require judicious application. The quality of evidence is graded as A (high quality randomized clinical trials), moderate (B) or low (C). This publication reports the key recommendations of the Writing Group. It is also accessible at http://www.bcshguide lines.com.

Management of acute anemia in a Jehovah's Witness patient with acute lymphoblastic leukemia with polymerized bovine hemoglobin‐based oxygen carrier: a case report and review of literature

Management of severe symptomatic anemia is exceptionally challenging when blood transfusions cannot be administered. Use of hemoglobin (Hb)-based oxygen carriers as "bridging treatment" can be an option. This is a report of a 36-year-old Jehovah's Witness diagnosed with acute lymphoblastic leukemia (ALL) who developed severe symptomatic anemia (Hb as low as 3.1 g/dL) during induction chemotherapy and refused allogeneic blood transfusions. Fifteen units of Hemopure (Biopure Corp.), an investigational polymerized bovine Hb-derived artificial oxygen carrier, were administered over a 12-day period after informed consent was obtained. The patient's condition improved. Chart review was performed to identify relevant information about the patient's clinical status during Hemopure treatment. With the use of Hemopure the patient's total Hb level was maintained between 3.6 and 5.3 g/dL, over a 12-day period, without any evidence of ischemia or organ dysfunction indicating that Hemopure was providing adequate tissue oxygen supply until marrow red blood cell recovery. The patient completed the chemotherapy regimen and was discharged in stable condition and has been in remission for 28 months. This report documents the successful management of profound anemia in an acute leukemia patient who refused blood transfusion, with the use of Hemopure, to save the patient from the devastating effects of ischemia associated with severe anemia. Hemopure may represent a life-saving intervention in patients with severe symptomatic anemia when blood transfusion is not an option.

Comparative study of predeposit and bedside leucodepletion filters

Background: Removal of leucocytes from cellular blood components is associated with reduction of several transfusion associated adverse reactions. Methods: A total of 400 units of packed red blood cells (RBCs) were subjected to leucodepletion at room temperature and 4°C using different commercially available prestorage and bedside filters (Terumo Penpol Immugard III and Pall Medical BPF-4). Prefiltration and post-filtration parameters were compared to assess the efficacy of prestorage leucodepletion vis-à-vis bedside leucodepletion and the requirement of universal leucodepletion. Result: Mean post-filtration red cell recovery ranged from 88.49-93.49% with all bags showing more than 85% red cell recovery. Mean post-filtration residual leucocyte count ranged from 0.205 x 10 6-0.338 x 10 6/bag with all bags showing more than log 3 leucoreduction. Prestorage leucoreduction achieved by the polyurethane filter was better than that achieved by the polyester filter. Red cell recovery with the bedside filters at room temperature was significantly less than that with prestorage filters at either temperature. Conclusion: This study suggests that prestorage leucoreduction is preferable over bedside leucoreduction and that polyurethane filters are better than polyester filters since leucodepletion achieved with the former is higher. We recommend selective log 3 leucodepletion using polyurethane prestorage filters for patients with specific indications.

High‐dose cyclophosphamide treatment for refractory severe aplastic anemia in children

To determine if high-dose cyclophosphamide is an effective therapy for children with refractory severe aplastic anemia (SAA). SAA is an illness characterized by the depletion of hematopoietic precursors associated with life-threatening complications. Hematopoietic stem cell transplant (HSCT) is the treatment of choice if a human leukocyte antigen (HLA)-related donor is available. Immunosuppression with anti-thymocyte globulin (ATG) and cyclosporine A (CSA) is an option for patients who are not HSCT candidates. Unrelated donor HSCT has been used with limited success. High-dose cyclophosphamide has been used successfully in the treatment of adults with SAA, but experience in children is limited. Five pediatric patients who had failed previous immunosuppressive therapy for SAA were treated with high-dose cyclophosphamide (45 mg/kg/day x 4 days). After 12 months of treatment, two of five patients experienced a complete response with high-dose cyclophosphamide therapy. The two complete responders achieved red cell recovery with a hematocrit of >36% at days 212 and 112 and platelet recovery with a platelet count of >100 x 10(9)/L at days 126 and 324. Of the remaining patients, one patient failed to respond, and two patients expired from infectious complications. High-dose cyclophosphamide can lead to complete responses in children with SAA who have failed to respond to traditional immunosuppressive therapy.

Rejuvenation of Aged Human Red Cells Improves Their in Vivo Recovery in a Mouse Model

Evaluation of novel storage or processing technology for human red blood cells (RBCs) involves in vitro tests on the red cells to determine biochemical changes and in vivo studies in healthy human volunteers with radiolabeled red cells to determine in vivo recovery 24 hours post infusion. In vivo studies are needed because our understanding of red cell storage lesions is not sufficient to identify an in vitro test(s) that would adequately predict red cell performance in vivo. The clinical studies with radiolabeled cells are used as the gold standard for evaluation prior to approval of a novel technology by the FDA. However, in vivo studies require time and funds and can be a significant hurdle in the development of new products. An animal model that could predict performance of human red cells in vivo would be useful in the development process. We previously reported that severe combined immunodeficient (SCID) mice could be used as a model to identify damaged human platelets (Transfusion. 47(8):1540–9, 2007). In the current study, we investigated if this murine model could be used to distinguish between the recovery of fresh and aged human RBCs, non-rejuvenated and rejuvenated aged RBCs, gamma-irradiated (25 Gy) fresh RBCs and irradiated fresh RBCs and stored for 28 days. “Fresh” RBCs were processed from whole blood within 24 hrs of collection and the “aged” RBCs were either RBC products stored for 42 or 100 days in an additive solution at 4°C. For in vivo recovery, approximately 1x109 human RBCs were injected into the tail vein of SCID mice (n=5 or 7 per condition) and serial blood samples were collected. Human RBCs were detected in mouse whole blood by flow cytometry using an anti-human glycophorin A mAb (clone CLB-ery-1). Recovery was defined as percent of human RBCs in the mouse circulation at 2 hours post infusion. Rejuvenation of cells was accomplished by incubating RBCs for 1 hour with Rejuvesol solution (Table 1). 2,3-DPG Levels (mM/L) Pre- and Post-Rejuvenation Fresh RBCs Aged for 42 Days Aged for 100 Days Control 3.25±0.40 0.17±0.04 0.38 ±0.06 Rejuvenated 8.58±0.82 4.56±0.17 2.31±0.13 Fresh red cells exhibited recovery of 58.4±4.4 % of total cells injected. Aged RBCs showed a reduced in vivo recovery of 35.7±7.3 % and 5.7±1.6 % of total cells injected for 42 and 100 day old RBC, respectively. Gamma-irradiated fresh RBCs and irradiated fresh RBCs stored for 28 days showed a recovery of 66.7±8.6 % and 55±13.2 % respectively, whereas the recovery of control fresh RBCs and control fresh RBCs stored for 28 days showed a recovery of 58.4±4.4 % and 49.1±7.0 % (p=0.44) respectively (Table 2). In VivoRecovery Fresh RBCs Stored for 28 days Aged for 42 Days Aged for 100 Days nd - not determined Control 58.4±4.5 49.1±7.0 35.7±7.3 5.17±1.6 Rejuvenated 52.5±11.5 nd 55.4±7.1 21.3±5.0 Irradiated (25Gy) 66.7±8.6 55±13.2 nd nd Our data indicate that the SCID mouse model can distinguish between fresh and aged red cells and that rejuvenation of the red cells increases intracellular 2,3-DPG levels and in vivo recovery of aged red cells. The SCID mouse model could be used to develop or improve existing methods of red cell storage and processing. The findings and conclusions in this abstract have not been formally disseminated by the Food and Drug Administration and should not be construed to represent any Agency determination or policy.

A comparison of hemolysis and red cell mechanical fragility in blood collected with different cell salvage suction devices

Suction pressure is one variable that can affect the efficiency of red blood cell (RBC) recovery for intraoperative autotransfusion. This study compared a constant-suction-pressure system with a device that is expected to minimize hemolysis by automatically varying the suction pressure. Twenty-two 50-mL reconstituted whole-blood samples were collected from a flat surface with either a constant-pressure suction device (BRAT 2 autologous blood recovery system, COBE Cardiovascular) set to a pressure of -200 mmHg or a variable-pressure suction device (SmartSuction Harmony surgical suction pump, Haemonetics, Inc.). Time of blood collection was recorded and plasma free hemoglobin (PFHb) was spectroscopically measured in the aspirated blood. Postcollection blood was subjected to RBC mechanical fragility test to characterize potential sublethal blood trauma. Relative hemolysis and the mean fragility index (MFI) were calculated. Hemolysis was significantly reduced with the variable-pressure suction compared to the constant-pressure suction (2.17% vs. 3.20%, respectively; p < 0.001). There was no significant difference between both suction devices in either the MFI of RBCs (0.632 vs. 0.673, respectively; p > 0.05) or the collection time at -200 mmHg (54.2 seconds vs. 52.8 seconds, respectively; p > 0.05). Although the variable-pressure device produced a significant reduction in hemolysis during one-pass blood collection, the clinical significance of this reduction is not clear. In relative terms, the variable-pressure device would recover an extra 10 mL of RBCs for every liter of salvaged RBCs, which is negligible compared to the blood loss in major surgery.

Evaluation of proposed FDA criteria for the evaluation of radiolabeled red cell recovery trials

FDA requirements for recovery of radiolabeled red blood cells (RBCs) 24 hours after autologous reinfusion in normal subjects have evolved over time. This study defined the ability of currently available RBC systems to satisfy the most recently proposed criteria. STUDY DESIGN METHODS: RBC recoveries were collected from US laboratories participating in clinical trials for RBC systems that have received FDA approval. Data were stratified for analysis into liquid-stored, gamma-irradiated, and frozen components. With statistical software, 24 individual samples were randomly selected with replacement from each stratum, repeating this for 5000 sample groups per stratum, to simulate experimental outcomes for each population. The percentage of sample groups that passed each and all of the proposed FDA criteria was determined. This procedure was repeated for recovery success thresholds of 75, 70, and 67 percent. A total of 941 RBC recoveries were obtained from 11 laboratories and 34 studies performed between 1990 and 2006 for 12 sponsors. While the criterion for the mean was almost always satisfied, the standard deviation (SD) criterion was more problematic. Causing most failures was the success threshold definition. Changing the success threshold from 75 to 70 percent or 67 percent increased the likelihood of meeting the requirement for all RBC types. The probability of passing the FDA-proposed criteria for currently FDA-approved products was poor. Changing the success threshold for an individual RBC recovery from 75 to 67 percent resulted in improved ability to meet this criterion for all three RBC types. This change had no affect on the pass rates based on the mean and SD criteria.

Neutrophil Gelatinase-Associated Lipocalin Levels in Patients with Thalassemia Intermedia: Lack of Association with Erythropoiesis, Iron Metabolism, Renal Dysfunction and Inflammation.

Neutrophil Gelatinase-Associated Lipocalin (NGAL, 24p3, SIP24, lipocalin-2 or siderocalin) is a novel iron-carrier protein exerting pleiotropic properties. The physiological role of NGAL is not fully elucidated though a few pivotal functions, such as regulation of apoptosis of leukocytes have been described recently. It has also been demonstrated that NGAL is abundantly expressed in erythroid progenitor cells inducing apoptosis and inhibiting their differentiation. NGAL is reduced in the erythroid cells of mice with acute anemia and injection of recombinant lipocalin-2 delayed the recovery of red blood cells suggesting a negative effect of NGAL in accelerating erythropoiesis. The recently reported functions of NGAL on erythroid progenitor cells and its relation to iron transport prompted us to study the pattern of NGAL levels in Thalassemia Intermedia (TI), which is characterized by erythroid progenitor cells hyperactivity and irregularities in iron metabolism (especially iron transport). To this end we measured NGAL levels in 30 patients with TI and in 20 apparently healthy controls matched for age. We also evaluated other parameters that could be possibly related to NGAL's functions as: soluble transferrin receptor (sTfR) for eythroid marrow activity; NTBI concentration for non-transferrin-bound mediate iron delivery pathway; creatinine, cystatin C and β2-microglobulin for renal dysfunction and CRP and IL-6 for low grade inflammation. The main results of the showed that: a) NGAL levels were significantly higher in patients with TI compared to controls (139.1±86.1 vs 51.2±11.8 μg/L, p<0.0001), while 4 patients presented with NGAL levels within normal range, b) in patients with TI no correlation was found between NGAL and the erythropoiesis' parameters Hb, Hb F and sTfR, (p>0.66, p>0.67 and p>0.81 respectively), the parameters of iron metabolism, ferritin and NTBI (p>0.90 and p>0.95 respectively) the markers of renal dysfunction and/or those of low-grade inflammation (p>0.50). The increased NGAL levels observed in TI patients in this study are in agreement with the elevated expression of NGAL observed in experimental models of thalassemia. We postulate that the induction of NGAL in these patients may represent either a response facilitating the survival of the less damaged thalassemic erythroid precursors, or a consequence due to the abnormal iron regulation in patients with TI.

The Mobilization Effects of G-CSF, GM-CSF and Darbepoetin-α for Allogeneic Peripheral Blood Stem Cell Transplantation.

Background: Granulocyte colony-stimulating-factor (G-CSF) is widely used for the mobilization of hematopoietic stem cells from normal donors. It has been reported that the addition of granulocyte-macrophage CSF (GM-CSF) to G-CSF mobilized more primitive CD34+ subsets than did G-CSF alone. We investigated the effects of combination regimen (GM-CSF + G-CSF + darbepoetin-α) for mobilization in normal healthy donors and compared its efficacy in mobilizing stem cells with G-CSF alone.Methods: Thirty donors were randomly assigned to one of the following regimens for mobilization: G-CSF group (G-CSF 5ug/kg/day for 5 to 7 days) and combination group (GM-CSF 10ug/kg/day on 1st and 2nd days, G-CSF 5ug/kg/day for 5 to 7 days and 40mg of dalbepoetin-α on day 1)Results: The counts of MNC and CD34+ cells/kg were not significantly different between two groups (p=0.129 and p=0.651). The CD3+ cell counts per kilogram were 3.48±1.45×108/kg and 2.54±0.94×108/kg in a G-CSF group and a combination group, respectively (p=0.04). Red blood cell transfusion requirement was not different between two groups (p=0.469). The median time to neutrophil and platelet engraftment was 12.5 vs. 12.0 days (p=0.443) and 11.5 vs. 11.0 days (p=0.752). Red blood cell recovery was 7.5 days and 7 days in G-CSF group and combination group, respectively (p=0.231). Grade II–IV acute graft-versus-host disease (GVHD) was observed in 20% in G-CSF group and 13% in combination group (p=0.50).Conclusion: The combination regimen was not superior to G-CSF alone in terms of acute GVHD and CD34+ cell dose. But CD3+ cell count was significantly lower in combination regimen group. The role of lower CD3+ cells in the graft need to be elucidated in the future.

Automated collection of double red blood cell units with a variable‐volume separation chamber

Automated collection of blood components offers multiple advantages and has prompted development of portable devices. This study sought to document the biochemical and hematologic properties and in vivo recovery of red cells (RBCs) collected via a new device that employed a variable-volume centrifugal separation chamber. Normal subjects (n = 153) donated 2 units of RBCs via an automated blood collection system (Cymbal, Haemonetics). Procedures were conducted with wall outlet power (n = 49) or the device's battery source (n = 104). Units were collected with or without leukoreduction filtration and were stored in AS-3 for 42 days. The units were assessed via standard biochemical and hematologic tests before and after storage, and 24 leukoreduced (LR) and 24 non-LR RBCs were radiolabeled on Day 42 with Na(2)(51)CrO(4) for autologous return to determine recovery at 24 hours with concomitant determination of RBC volume via infusion of (99m)Tc-labeled fresh RBCs. Two standard RBC units (targeted to contain 180 mL of RBCs plus 100 mL of AS-3) could be collected in 35.7 +/- 2.0 minutes (n = 30) or 40.3 +/- 2.7 minutes for LR RBCs (n = 92). An additional 31 collections were conducted successfully with intentional filter bypassing. RBC units contained 104 +/- 4.1 percent of their targeted volumes (170-204 mL of RBCs), and LR RBCs contained 92 percent of non-LR RBCs' hemoglobin. All LR RBCs contained less than 1 x 10(6) white blood cells. Mean hemolysis was below 0.8 percent (Day 42) for all configurations. Adenosine triphosphate was well preserved. Mean recovery was 82 +/- 4.9 percent for RBCs and 84 +/- 7.0 percent for LR RBCs. The Cymbal device provided quick and efficient collection of 2 RBC units with properties meeting regulatory requirements and consistent with good clinical utility.

Granulocyte and erythropoietic stimulating proteins after high-dose chemotherapy for myeloma

High-dose chemotherapy is an established treatment for patients with myeloma. In randomized trials it has been shown to prolong disease-free survival by around 1 year compared to patients receiving chemotherapy alone. Physically and psychologically high-dose therapy takes its toll on the patient who may be in hospital for around 3 weeks and take some weeks or months to convalesce after discharge. Granulocyte colony stimulating factors and erythropoietic stimulating agents will speed neutrophil and red cell recovery, respectively, when used at an appropriate time after the high-dose chemotherapy. The clinical value of these laboratory findings is uncertain and the role of these agents after high-dose chemotherapy remains a subject for debate.

In vivorecovery and survival of red cells after photodynamic treatment with thiopyrylium and red light using a canine model

Recently, we have shown that thiopyrylium has robust inactivation capabilities against a broad spectrum of pathogens in the human red blood cell (RBC) suspensions while retaining key RBC in vitro properties. The 24-h recovery and survival of canine red cells were measured upon autologus reinfusion of control and phototreated units. The 24-h recovery of control and phototreated RBCs was 75.7 +/- 6.4% and 87.5 +/- 8.5%, respectively. The time for 50% survival of labelled control RBCs was similar to that of phototreated group (206 +/- 58 h vs. 255 +/- 63 h, respectively). Results suggest that thiopyrylium phototreatment does not negatively affect canine RBC in vivo recovery.

The Post-Transfusion Recovery and Survival of Red Blood Cells in Mice Is Affected by the Expression of Cellular Prion Protein.

Three documented transfusion cases of vCJD underline the need of better insight in blood prion protein biology. Cellular prion protein (PrPc) plays key role in the pathophysiology of prion diseases. Its expression by cells is necessary for amplification of infectious prions and the disease process itself. Physiological function of PrPc remains obscure. Its clarification may provide important clues for the development of urgently needed blood test and effective disease treatment. PrPc is expressed on CD34+ hematopoietic stem cells and its expression is regulated during blood cell differentiation. Recently the importance of PrPc for self-renewal of long-term repopulating hematopoietic stem cells was suggested and other studies reported the protective function of PrPc against oxidative stress and apoptosis in various cell cultures. We previously demonstrated that human as well as mouse red blood cells (RBC) express approximately 200 PrPc molecules / cell (Holada et al., BJH 2000, 110, 472–80). To test if the PrPc expression plays a role in the post-transfusion recovery and survival of RBC we carried out transfusion study in mice. RBC isolated from blood of wild type (WT) and PrP knockout (KO) FVB mice were labeled “in vitro” by different levels of NHS-biotin. The labeling was optimized to allow simultaneous detection of both populations of RBC in mouse blood using flow cytometry. To exclude the influence of different level of cell biotinylation on the experiment outcome two mixtures of RBC were prepared. The first contained KO RBC labeled with high and WT RBC with low level of biotin and the second mixture contained cells labeled “vice versa”. Each mixture was injected via tail vein in a group of WT mice (n=5) and the survival of RBCs was followed. Samples were analyzed on day 1, 2, 3, 6, 9, 15, 21 and 29. The count of biotinylated RBC was measured in comparison to 100 000 nonlabeled recipient RBC. Simultaneously the expression of PrPc on RBC was monitored using flow cytometry with MAb 6H4. KO RBC displayed significantly higher first day post-transfusion recovery compared to WT RBC in both groups of mice (81 ± 3 % vs. 74 ± 3 %, P<0.005 and 90 ± 4 % vs. 80 ± 4 %, P<0.005). The slope of the RBC survival curve in all individual mice during the initial 15 days was steeper for KO RBC (mavg = − 3.44) than for WT RBC (mavg = − 2.37) suggesting the protective role of PrPc in circulating RBC. The difference in the slope diminished during the 15 to 29 day period which was accompanied by a 50% decrease of PrPc surface expression on transfused WT RBC. To confirm our data the identical experiment was carried out in a group of KO mice (n=5) transfused with a mixture containing KO RBC labeled with low and WT RBC with high level of biotin. Again the first day post-transfusion recovery was higher for KO RBC (80 ± 6 % vs. 75 ± 6 %, P<0.05) and the initial slope of the KO RBC survival curve was steeper in all mice in the group. Our data suggest that PrPc expression plays role in the post-transfusion recovery and survival of RBC. The observation that WT RBC disappear from the circulation at lower rate than KO RBC until their level of surface PrPc reaches 50% is compatible with the protective role of PrPc expression on cells. Taken together our study demonstrates that physiological role of PrPc expression on RBC may lay in facilitating their longer survival in circulation. (GACR 310/04/0419, MSMT 0021620806).

Prestorage leucoreduction improves several in vitro red cell storage parameters following gamma irradiation

Gamma irradiation (GI) can prevent transfusion-associated graft-vs.-host disease, result in a small decrement in 24-h in vivo red blood cell (RBC) recovery and in a minor loss of maintenance of several in vitro parameters during routine storage. Results from studies by Davey et al. (Transfusion 1995, 35, 55S) demonstrated that the decrement in 24-h recovery was not observed if units were first leucoreduced (LR), suggesting that RBC damage on leucocytes from GI was secondary to the effects of irradiation . In this study, we further investigated how leucoreduction affected eight in vitro storage parameters of gamma-irradiated units by using a matched sample study design. Leucoreduction significantly reduced (P < 0.05) the deleterious effect of GI on adenosine triphosphate glucose levels, haemolysis and mean corpuscular volume (MCV) for AS-3 units and significantly decreased (P < 0.05) the deleterious effect of GI on glucose levels, haemolysis and MCV for AS-1 units. Leucoreduction did not influence (P >> 0.5) the enhanced potassium release associated with GI. Some, but not all, the observed RBC damage caused by GI appears to be secondary to the effect of irradiation on leucocytes. Based on the in vivo survival studies of Davey et al. and the results of this study, there is justification to consider performing additional survival studies to support extended storage of LR gamma-irradiated RBC.

Practice Guidelines for Perioperative Blood Transfusion and Adjuvant Therapies

Practice Guidelines for Perioperative Blood Transfusion and Adjuvant Therapies

Adhesion receptor expression by CD34 + cells from peripheral blood or bone marrow grafts: Correlation with time to engraftment

This study was undertaken to define the pattern of cell adhesion receptor expression by the CD34+ progenitor cells from mobilized peripheral blood and bone marrow from normal and autologous donors, and to correlate the adhesion receptor profile with time to blood cell recovery for patients undergoing autologous transplant. Blood cell recovery was determined by absolute neutrophil count (> 500/microL), time to last red cell transfusion, and platelet count (> 50,000/microL and > 100,000/microL). The analysis for expression of adhesion receptors alphaL (CD11a), alpha2 (CD49b), alpha3 (CD49c), alpha4 (CD49d), alpha5 (CD49e), alpha6 (CD49f), beta1 (CD29), L-selectin (CD62L), ICAM-1 (CD54), PECAM-1 (CD31), HCAM (CD44), and P-selectin (CD62P) was performed by two-color flow cytometry. The Kruskal-Wallis test and Spearman's rank correlation were used for statistical analysis. Statistical analysis of adhesion expression by the CD34+ population for patients undergoing autologous transplant demonstrated that the higher percent expression of PECAM-1 correlated with longer time to platelet recovery (> 100,000/microL, p = 0.049). In contrast, the higher the percent expression of alpha6 (p = 0.013) and the increased density of expression of alpha2 (p = 0.035), alpha3 (p = 0.023), alpha4 (p = 0.044), beta1 (p = 0.027), and ICAM-1 (p = 0.010) correlated with shorter time to platelet recovery. Neutrophil recovery time decreased with increased density of expression of alphaL (p = 0.014) and P-selectin (p = 0.007) receptors. Increased density of expression of CD44 (HCAM) was associated with longer time to red blood cell recovery (p = 0.05). These data suggest that upregulation of specific adhesion receptors or selection of certain cell populations could result in earlier blood cell recovery after transplant.

Partial Bowls Using the Haemonetics Cell Saver 5: Does It Produce a Quality Product?

Controversy still exists on the validity of processing a partial bowl during the collection of shed blood lost through surgery during cell salvaging. The purpose of this study was to assess the quality of red blood cells produced from a partial bowl of autologous suctioned blood using the Haemonetics Cell Saver 5. Suctioned blood was collected from 17 patients undergoing cardiac surgery. A partially filled cell saver bowl was washed with 1500 mL of NaCl. Reservoir and processed blood samples were examined for potassium, leukocytes, hematocrit, platelets, and plasma-free hemoglobin and then compared with 22 previously studied full bowls. Results are summarized in the table below:In conclusion, the Haemonetics Cell Saver 5 can produce a quality product from washing a partial bowl with a better washout of white blood cells compared with a full bowl. However, there is a reduction in red blood cell recovery.

Does washing swabs increase the efficiency of red cell recovery by cell salvage in aortic surgery?

We investigated the contribution of swab washing to the efficiency of red cell recovery by intraoperative cell salvage (ICS) in 10 patients undergoing elective aortic aneurysm repair. Volumes and haemoglobin (Hb) concentrations were recorded in the blood recovered by direct suction and from washed swabs, both before and after processing with a Haemonetics Cell Saver 5. The mean +/- standard deviation (SD) estimated blood loss was 991 +/- 403 ml, resulting in a mean +/- SD salvaged RBC volume of 380 +/- 124 ml. The median [interquartile (IQR) range] Hb collected from suction was 84.9 (61.8-131.4) g, of which 50.1 (45-71.5) g was returned to the patient after processing, a median yield of 68 (49-77)%. The swab wash produced a median (IQR) Hb of 39.4 (28.4-64.9) g, of which 26.2 (16.8-31) g was reinfused, a 67 (33-98)% yield. Swab wash thus contributed with a median (IQR) of 31 (24-39)% of the total RBC recovery. Washing swabs improves the efficiency of red cell recovery by ICS.

Effect of different anesthesia techniques on red blood cell endogenous recovery in hip arthroplasty

Study Objective To compare the magnitude of postoperative red blood cell (RBC) recovery with 3 different anesthetic techniques, general anesthesia (GA), epidural anesthesia (EA) alone, and the combination of these 2 techniques (CA), in patients undergoing total hip arthroplasty. Design Randomized, controlled study. Setting Seven university or hospital departments of anesthesia. Patients Two hundred ten patients with American Society of Anesthesiologists physical status I to III were randomly selected to receive EA, GA, or CA. Intervention Patients undergoing total hip replacement were randomly assigned to 3 statistically comparable groups based on the type of anesthesia received: GA, EA, and CA groups. Measurements and Main Results Intra- and postoperative blood loss was evaluated as either compensated or noncompensated blood loss by using Nadler's formula. The intra- and postoperative bleeding, referred to as compensated blood loss, was similar among groups. The circulating RBC mass, noncompensated blood loss, dropped on the first postoperative day to a similar extent among the groups. The endogenous recovery of the RBC is carried out on the fifth day after surgery in patients who underwent EA, whereas no RBC recovery was observed in those who had received GA alone or GA combined with EA. Conclusions Patients who had received EA had a faster recovery of the circulating erythrocyte mass than those who had had GA or CA. The presence of nitrous oxide in the anesthetic gas mixture might inhibit erythropoiesis by altering vitamin B 12 functions.