bluuebunny/crossref_metadata_2025 huggingface dataset:
Prevalence of hepatitis C virus and genotype distribution in an Australian volunteer blood donor population
Authors: Mison L.M. , Young I.F. , O'Donoghue M. , Cowley N. , Thorlton N. , Hyland C.A.
Published: March 2003

BACKGROUND: This study was undertaken to assess the prevalence of hepatitis C virus (HCV) antibody and RNA in first‐time blood donors and to examine the HCV genotype distribution.

STUDY DESIGN AND METHODS: A third‐generation enzyme‐linked immunosorbent assay (ELISA) was used to screen 34,725 donors for HCV antibodies. Donors who were repeatably reactive were tested in two immunoblot assays—a second‐generation and a third‐generation recombinant immunoblot assay—as well as by a polymerase chain reaction (PCR) assay. PCR‐positive donors were genotyped. All samples were screened for alanine aminotransferase levels.

RESULTS: The ELISA repeat reactivity rate was 0.55 percent. PCR testing showed that 69 (38%) of the 183 ELISA‐reactive samples contained HCV RNA. The third‐generation recombinant immunoblot assay identified all 69 viremic samples as antibody positive; however, only 63 tested positive on the second‐generation immunoblot. The remaining six PCR‐positive donors tested antibody‐indeterminate to the core peptide. All six of these donors had HCV subtype 3a infections. Genotype distribution among 58 samples showed that 34 were type 1, of which 22 could be further subtyped as 1a (16) and 1b (6); 2 were 2a; 5 were 2b; and 17 were subtyped as 3a. Donors infected with 2b and 3a had reduced antibody reactivity to the NS4 and NS3 peptides only on the second‐generation immunoblot.

CONCLUSION: The prevalence of confirmed anti‐HCV and viral RNA in new donors is 0.29 and 0.2 percent, respectively. The third‐generation recombinant immunoblot assay was more sensitive than the second‐generation immunoblot assay in detecting 2b and 3a HCV subtypes. The inclusion of the NS5 peptide in the third‐ generation recombinant immunoblot did not result in positive tests in any additional donors. Rather, the improvement was due to the increased detection of NS3 and, to a lesser extent, NS4 antibodies. Subtypes 1a and 3a were most prevalent in this population.
Improvement of platelet storage conditions by using new polyolefin containers
Authors: de Wildt‐Eggen J. , Schrijver J.G. , Bouter‐Valk H.J. , Fijnheer R. , Bins M. , van Prooijen H.C.
Published: March 2003

BACKGROUND: Storage of pooled platelet concentrates (PCs) with yields above 3.0 × 1011 platelets per unit in a 1‐L PL‐732 polyolefin container for 5 days often results in a drop in pH to below 6.0. Recently, new oxygen‐permeable platelet containers (1‐L PL‐2410, 1‐L and 1.5‐L Compoflex) have been developed. The maximal platelet storage capacities of the new containers and the PL‐732 were compared.

STUDY DESIGN AND METHODS: Large platelet pools (n = 27) with platelet concentrations between 1.2 and 1.4 × 1011 per L were made from 3 to 5 PCs prepared from buffy coats. The pools were divided in equal volumes among the PL‐732 and the three new platelet containers. Platelet counts in the PCs ranged from 1.0 to 5.0 × 1011 per unit. All PCs were stored on a flatbed shaker at 22 ± 2°C and evaluated on Days 1, 3, 5, and 7 by measuring platelet count, pH, pO2, pCO2, HCO3‐, glucose, lactate, platelet swirling, and soluble p‐selectin.

RESULTS: Day 7 storage of PCs (n = 6) with yields between 3.0 and 4.0 × 1011 platelets in PL‐732 showed mean ± SD pH values of 5.93 ± 0.05 and lactate values of 32.3 ± 7.9 mmol per L; in 4 of these 6 PCs, pH was below 6.0. In contrast, storage of these PCs in 1‐L PL‐2410 and 1.5‐L Compoflex containers and of 2 of these 6 PCs in 1‐L Compoflex containers showed pH values above 6.8. Lactate values were 15.5 ± 1.3, 15.3 ± 1.8, and 19.5 ± 4.7 mmol per L, respectively (p < 0.001 vs. PL‐732). The platelet storage capacity of the new containers with platelet yields between 4.0 and 5.0 × 1011 per unit (n = 6) was evaluated. Day 7 storage of these PCs in the 1.5‐L Compoflex showed an average pH value of 6.74 ± 0.20; in 2 of 6 PCs, pH was below 6.8. The average pH value in the PL‐2410 was 6.38 ± 0.31, and in all PCs, pH was below 6.8. Average lactate values were 17.8 ± 5.7 and 25.8 ± 5.6 mmol per L (p < 0.05), respectively. Soluble p‐selectin values on Day 7 of storage increased approximately twofold in all PCs.

CONCLUSION: The new oxygen‐permeable containers showed platelet quality comparable to that with the PL‐732 and for longer storage periods and at higher platelet counts.
Early changes in hemoglobin and hematocrit levels after packed red cell transfusion in patients with acute anemia
Authors: Elizalde J. Ignasi , Clemente Josefina , Marín José Luis , Panés Julian , Aragón Blanca , Mas Antoni , Piqué Josep M. , Terés Josep
Published: March 2003

BACKGROUND: Equilibration of hemoglobin concentration after transfusion has been estimated to take about 24 hours, but some studies have shown that earlier measurements reflect steady‐state values in persons who have not bled recently. This study was aimed at assessing the changes over time in hemoglobin concentration after transfusion in acutely anemic patients because of recent bleeding. STUDY DESIGN AND METHODS: Thirty‐two normovolemic patients recovering from an acute bleeding episode who were no longer thought to be bleeding and who received a 2‐ unit red cell transfusion were studied. At baseline and 15, 30, 60, and 120 minutes and 24 hours after transfusion, hemoglobin concentration and hematocrit values were measured. RESULTS: The administration of 2 units of packed red cells elicited a 24‐hour increase of 22.4 +/− 6.8 g per L in hemoglobin concentration. Hemoglobin values were not different at any of the defined posttransfusion times. Hematocrit levels experienced similar changes over time. Agreement between 15‐minute and 24‐hour values was excellent, as only 6 percent of patients exhibited a clinically significant difference (> 6 g/L) between the hemoglobin measurements. CONCLUSION: Hemoglobin and hematocrit values rapidly equilibrate after transfusion in normovolemic patients who are recovering from an acute bleeding episode. This fact would allow a rapid assessment of the effects of transfusion and of the recurrence of bleeding in patients remaining at risk.
Recombinant human erythropoietin as adjuvant treatment for autologous blood donation in elective surgery with large blood needs (> or=5 units): a randomized study
Authors: De Pree Christian , Mermillod Bernadette , Hoffmeyer Pierre , Beris Photis
Published: March 2003

BACKGROUND: Autologous blood transfusion presents no infectious or immunologic side effects. The aim of this randomized study was to determine the impact of recombinant human erythropoietin (rHuEPO) on the donation of 5 units of autologous blood by nonanemic patients who were candidates for elective surgery with transfusion requirements of > or=5 units. STUDY DESIGN AND METHODS: Starting on Day −35, 420 mL of blood was taken weekly. All patients received 200 mg of iron saccharose complex intravenously at each visit and six subcutaneous injections of rHuEPO (141 U/kg) or placebo between Days −21 and −7. RESULTS: Of 50 patients, 45 completed the study (placebo, 21; rHuEPO, 24). Total red cell production was higher in the rHuEPO group (p=0.001). Donation of 5 units was possible for 67 percent (placebo group) and 79 percent (rHuEPO group) of patients (p=0.5). The mean number of blood units donated was 4.6 (placebo group) and 4.7 (rHuEPO group). More patients in the placebo group received allogeneic blood (9/21 [43%] vs. 6/23 [26%]), although the difference did not reach significance (p=0.34). CONCLUSION: In nonanemic patients donating 5 units of blood, rHuEPO associated with intravenous iron increased total red cell production. However, no difference was found between the rHuEPO and placebo groups with regard to the number of units of autologous blood donated of the number of patients receiving allogeneic blood transfusion.
Peripheral blood CD34+ cell immunomagnetic selection in breast cancer patients: effect on hematopoietic progenitor content and hematologic recovery after high‐dose chemotherapy and autotransplantation
Authors: Cancelas J.A. , Querol S. , Canals C. , Picón M. , Azqueta C. , Solà C. , Montes A. , Amill B. , Griera E. , Inglés J.R. , López J.J. , Germà J.R. , García‐López J.
Published: March 2003

BACKGROUND: Tumor cells have been detected in mobilized peripheral blood of breast cancer patients, and they may contribute to tumor recurrence after the transplantation of peripheral blood progenitor cells. One of the most widespread technologies for tumor purging of the graft is immunomagnetic hematopoietic progenitor cell selection. STUDY DESIGN AND METHODS: The study assessed the effectiveness of a magnetic cell‐separation system in selecting functional subpopulations of hematopoietic progenitors from 14 blood‐derived harvests of 11 patients with high‐risk breast cancer after mobilization following cytotoxic chemotherapy supported by granulocyte–colony‐stimulating factor, as well as the feasibility of transplanting these selected subpopulations. RESULTS: CD34(+)‐enriched cell fractions had a median purity of 93.0 percent (72.7‐98.5%). The procedure yielded 52.6 percent of the CD34+ cell input (39.4‐116.8%). Median recoveries of colony‐forming units (CFUs) (36.87%) and cobblestone area‐forming cells (CAFCs) (152.5%) were, respectively, 0.70 and 2.87 times those of CD34+ cells (52.6%). Moreover, CAFC efficiency in the positive cell fraction was 2.57 times that in the starting cell fraction. Peripheral blood neutrophil counts of 0.5 × 10(9) per L and platelet counts of 20 × 10(9) per L were reached after median times of 9 and 11 days, respectively. The number of transfused CAFCs per kg, CD34+ cells per kg, and postthaw CFU‐ granulocyte‐macrophage per kg was correlated, respectively, with the speed of engraftment of neutrophils, platelets, or both. Tumor cells detected in one patient's peripheral blood were not found after CD34+ cell selection. CONCLUSION: Transplantation of immunomagnetically purified peripheral blood CD34+ cells does not increase transplantation‐ related morbidity. It induces a selective enrichment of more immature hematopoietic progenitors, which makes it suitable for use in cell expansion and gene therapy protocols.
Saponin, an inhibitory agent of carbon dioxide production by white cells: its use in the microbiologic examination of blood components in an automated bacterial culture system
Authors: van Doorne H. , van der Tuuk Adriani W.P.A. , van de Ven L.I. , Bosch E.H. , de Natris T. , Sibinga C.T. Smit
Published: March 2003

BACKGROUND: Blood components with a white cell count > 100 × 10(9) per L may cause false‐positive results when the BacT/Alert system is used for the microbiologic examination. The effects of different concentrations of saponin on bacterial growth and on carbon dioxide production by blood fractions with a high white cell count, in particular peripheral blood progenitor cells and buffy coats, were investigated. STUDY DESIGN AND METHODS: The effect of saponin on carbon dioxide production was studied by adding different fractions of white cell‐rich material (buffy coat or leukapheresis material) to BacT/Alert culture bottles with or without saponin and incubating these bottles. Five bacterial strains were used to inoculate the culture bottles at four levels ranging from about 1 colony‐forming unit per mL to about 10(3) colony‐forming units per mL. Aerobic and anaerobic bottles with and without saponin were used. RESULTS: It was demonstrated that the addition of 0.5 percent saponin to BacT/Alert culture bottles effectively inhibited carbon dioxide production, without affecting bacterial growth. CONCLUSION: Saponin at a concentration of 0.5 percent is a valuable additive to BacT/Alert culture media because it prevents false‐positive results in the examination of white cell‐rich blood components.
Transforming growth factor beta is increased in plasma of patients with hematologic malignancies after transfusion of platelet concentrates
Authors: Kunz D. , Luley C. , Heim M.U. , Bock M.
Published: March 2003

BACKGROUND: Transforming growth factor beta 1 (TGF‐beta 1) acts as a potent inhibitor of bone marrow proliferation. High concentrations were found in human platelets, which release this cytokine during storage. STUDY DESIGN AND METHODS: TGF‐beta 1 levels during a storage period of 5 days were compared in the plasma of platelet concentrates prepared by apheresis or by the buffy coat method. In addition, TGF‐beta 1 plasma levels were monitored in patients with hematologic malignancies before and after transfusion. RESULTS: TGF‐beta 1 levels in the supernatant of platelet concentrates were found to be 55 times higher than those in the plasma of healthy volunteer donors. During storage, an additional increase was observed. Accordingly, the transfusion of platelet concentrates resulted in a significant increase of plasma TGF‐beta 1 levels in patients with hematologic malignancies (before transfusion: 2.2 +/− 0.5 ng/mL; after transfusion: 2.9 +/− 0.6 ng/mL), and these higher levels persisted for at least 4 hours. CONCLUSION: Because TGF‐ beta 1 reduces the clonogenic capacity of hematopoetic progenitor cells, a myelosuppressive effect of platelet transfusions is suggested.
All adverse events in autologous blood donors with cardiac disease are not necessarily caused by blood donation
Authors: Kasper S.M. , Ellering J. , Stachwitz P. , Lynch J. , Grunenberg R. , Buzello W.
Published: March 2003

BACKGROUND:Autologous blood donation before elective cardiac surgery has become a standard of care at many institutions. However, the safety of autologous blood donations by patients with cardiac disease is subject to controversy. CASE REPORTS: Two life‐threatening cardiac arrests and one fatal myocardial infarction that occurred in three patients who were scheduled to donate blood for autologous use in elective cardiac surgery are reported. All three patients met the institution's selection criteria for autologous blood donors, and all of them had given written informed consent for their participation in the autologous blood donation program. One of the two cardiac arrests and the myocardial infarction occurred in the patients prior to any blood donations, and the other cardiac arrest occurred 7 days after the patient donated blood uneventfully. CONCLUSION: Life‐threatening and fatal adverse events may occur during the donation period in autologous blood donors with cardiac disease. Not all adverse events are necessarily caused by blood donation.
Efficacy of oral iron supplementation is not enhanced by additional intravenous iron during autologous blood donation
Authors: Kasper S.M. , Lazansky H. , Stark C. , Klimek M. , Laubinger R. , Borner U.
Published: March 2003

BACKGROUND: The study compared the efficacy of oral iron combined with intravenous iron supplementation to that of oral iron supplementation alone in increasing the preoperative production of hemoglobin (Hb) in autologous blood donors with normal iron stores.

STUDY DESIGN AND METHODS:One hundred eight iron‐replete patients who were scheduled for donation of 3 units of autologous blood at weekly intervals were randomly assigned to receive, in a double‐blind fashion, no iron supplementation (placebo, Group 1), oral iron supplementation (285.6 mg of elemental iron/day, Group 2), or oral iron plus intravenous iron supplementation (285.6 mg of elemental iron/day orally plus 102.5 mg of elemental iron/week intravenously, Group 3). The amount of Hb produced during the 21‐day study period was determined by the total amount of Hb donated minus the change in the amount of circulating Hb between the first donation (Day 0) and the poststudy examination (Day 21).

RESULTS:Hb production did not differ significantly in the two iron‐supplemented groups (oral iron, 85 +/− 36 g; oral plus intravenous iron, 74 +/− 43 g). The patients in the oral iron group produced a significantly greater amount of Hb than those in the placebo group (85 +/− 36 g vs. 52 +/− 41 g, p < 0.01). CONCLUSION: Oral iron supplementation increased the production of Hb in autologous blood donors more than placebo did. Additional intravenous iron did not lead to a further increase in preoperative Hb production.
White cell reduction during plateletpheresis: a comparison of three blood cell separators
Authors: Moog R. , Müller N.
Published: March 2003

BACKGROUND: White cell (WBC)‐reduced single‐donor platelet concentrates (SDPs) can be collected by the newest generation of blood cell separators. Three WBC‐reduction techniques during plateletpheresis were investigated in the present study with respect to WBC content and platelet yield.

STUDY DESIGN AND METHODS: The Amicus device used the elutriation principle for WBC reduction, and separations with periodically alternating interface position (PAIP) were employed in the AS.TEC 204. WBC reduction by in‐line filtration was performed in the MCS+. Platelets were measured electronically and WBCs were determined manually (Nageotte chamber).

RESULTS: In‐line filtered SDPs showed significantly lower WBC content (0.088 ± 0.178 × 106) than SDPs that were WBC reduced by elutriation (0.31 ± 0.48 × 106) or PAIP technique (0.89 ± 1.57 × 106, p = 0.0001). Platelet yield (5.0 ± 0.46 × 1011) was significantly higher in components obtained with the Amicus device (p = 0.0001). The AS.TEC 204 and MCS+ gave similar results for platelet yields: 3.15 ± 0.63 and 3.28 ± 0.71 × 1011, respectively.

CONCLUSIONS: The plateletpheresis systems studied allow the collection of WBC‐reduced SDPs. In‐line filtration resulted in the best WBC reduction. Some SDPs collected with the devices studied had a WBC content >1 × 106 per unit. Platelet yield was significantly higher in SDPs from the Amicus device.
FcγRIIIB polymorphism: evidence that NA1/NA2 and SH are located in two closely linked loci and that the SH allele is linked to the NA1 allele in the Danish population
Authors: Steffensen R. , Gülen T. , Varming K. , Jersild C.
Published: March 2003

BACKGROUND: The neutrophil‐specific antigens NA1, NA2, and SH are well‐recognized allotypic forms of FcγRIIIB. Individuals carrying all three FcγRIIIB genes were recently described.

STUDY DESIGN AND METHODS: A Danish population (n = 200) was typed for NA1, NA2, and SH by polymerase chain reaction with sequence‐specific primers (PCR‐SSP). Twelve individuals with three FcγRIIIB genes were further genotyped by PCR‐based restriction fragment length polymorphism and by DNA sequencing. Family studies were performed on three individuals who carry three FcγRIIIB genes.

RESULTS: The gene frequencies for NA1, NA2, and SH were 0.365, 0.635, and 0.030, respectively. In eight individuals (4%), all three FcγRIIIB genes were identified. All 12 SH+ individuals were NA1+.

CONCLUSION: The NA1, NA2, and SH gene frequencies observed in Danes are similar to those in other white populations. The distribution of FcγRIIIB genotypes in the Danish population strongly indicates that the NA and the SH systems are located in two closely linked loci and that SH is closely linked to NA1. Finally, a new PCR‐SSP was developed to distinguish NA2 from SH.
The viability of autologous human red cells stored in additive solution 5 and exposed to 25°C for 24 hours
Authors: Reid T.J. , Babcock J.G. , Derse‐Anthony C.P. , Hill H.R. , Lippert L.E. , Hess J.R.
Published: March 2003

BACKGROUND: No data exist on the viability of red cells (RBCs) storsed in modern additive solution systems and allowed to warm above 10°C.

STUDY DESIGN AND METHODS: In a randomized crossover study, 3 units of blood were collected at least 8 weeks apart from 11 volunteer donors and stored in additive solution 5 (AS‐5). Of 3 units from each volunteer, 1 was stored for 6 weeks at 4°C, 1 for 5 weeks at 4°C except for 24 hours at 25°C on Day 14, and 1 for 5 weeks at 4°C except for 24 hours at 25°C on Day 28. Units were sampled periodically during storage; at the end of storage, viability was measured by the 99mTc/51Cr double‐label method.

RESULTS: RBC viability was not significantly different in the storage protocols. Less than 1 percent of stored cells hemolyzed. RBC ATP concentrations at the end of storage correlated with viability and were approximately equal in the warmed units after 30 days' storage and the conventionally stored units after 42 days.

CONCLUSIONS: The data suggest that RBCs stored in AS‐5 and allowed to warm to 25°C for 24 hours lose about 12 days of their shelf life.
The influence of citrate concentration on the quality of plasma obtained by automated plasmapheresis: a prospective study
Authors: Beeck H. , Becker T. , Kiessig S.T. , Kaeser R. , Wolter K. , Hellstern P.
Published: March 2003

BACKGROUND: There is a need for more comprehensive work dealing with the quality of plasma collected by automated plasmapheresis using different final concentrations of citrate anticoagulant. A prospective study was performed to examine the influence of three concentrations of sodium citrate on the levels of clotting factors and markers of activated hemostasis and fibrinolysis.

STUDY DESIGN AND METHODS: Fifty‐one experienced plasma donors were recruited for subsequent 750‐mL plasmapheresis procedures using 4‐percent (wt/vol) sodium citrate. Anticoagulant‐to‐blood ratios of 1:16.6, 1:14.2, and 1:12.5 were used, corresponding to sodium citrate concentrations of 6 percent, 7 percent, and 8 percent (vol/vol), respectively. Between two plasmapheresis procedures, there was a washout period of 7 days. Determinations were made of the plasma levels of fibrinogen and factors V, VII, VIII, and IX, as well as antithrombin, tissue‐type plasminogen activator, and several markers of activated hemostasis and fibrinolysis: activated factor VII, prothrombin splits products, d‐dimers, and β‐thromboglobulin.

RESULTS: The plasma samples anticoagulated with 6‐percent citrate contained significantly higher levels of factors V, VIII, and IX than the samples anticoagulated with 8‐percent citrate (p<0.0001, p≤0.0001 and p = 0.009, respectively). The citrate concentration had no influence on the levels of fibrinogen, factor VII, antithrombin, or tissue‐type plasminogen activator. There was no evidence that the plasma samples containing lower citrate concentrations were more prone to activation of hemostasis or fibrinolysis.

CONCLUSION: A reduction in the final citrate concentration of plasma collected by automated plasmapheresis results in higher yields of factors V, VIII, and IX without activation of hemostasis. More comprehensive studies should confirm previous work dealing with the establishment of the lowest citrate concentration acceptable in plasma used as therapeutic fresh‐frozen plasma or as starting material for the manufacture of plasma derivatives.
Lookback on donors who are repeatedly reactiveon first‐generation hepatitis C virus assays:justification and rational implementation
Authors: Tobler L.H. , Tegtmeier G. , Stramer S.L. , Quan S. , Dockter J. , Giachetti C. , Busch M.P.
Published: March 2003

BACKGROUND: The purpose of this study was to explore strategies to minimize the number of unwarranted consignee notifications resulting from hepatitis C virus (HCV) first‐generation (single‐antigen) enzyme immunoassay (EIA 1.0) targeted lookback.

STUDY DESIGN AND METHOD: The four blood centers participating in this study contributed data on 3753 HCV EIA 1.0‐repeatably reactive (RR) donations. The analysis focused on 1) statistical evaluation of HCV EIA 1.0 signal‐to‐cutoff (S/CO) ratios versus HCV second‐generation recombinant immunoblot assay (RIBA 2.0) interpretation from all participating blood centers and 2) RNA testing using transcription‐mediated amplification on all HCV EIA 1.0 RR/RIBA 2.0‐positive or ‐indeterminate specimens and a subset of RIBA 2.0‐negative donations for which specimens were available.

RESULTS: Analysis of HCV EIA 1.0 S/CO ratios versus RIBA 2.0 indicated that 1180 (89%) of 1326 RIBA 2.0‐positive specimens had an S/CO ratio >2.5, while 146 (11%) had a ratio ≤2.5. In contrast, of 2253 RIBA 2.0‐negative specimens, 299 (13%) had an S/CO ratio >2.5, while 1954 (87%) had a ratio ≤2.5. Of 248 HCV EIA 1.0‐RR/RIBA 2.0‐positive samples with stored specimens available for additional testing, 198 (80%) were HCV RNA positive; 15 (7.5%) of these specimens had an S/CO ratio ≤2.5, while 183 (92%) had a ratio >2.5. HCV RNA was detected in only 2 (1.5%) of 137 HCV EIA 1.0‐RR/RIBA 2.0‐negative specimens: 1 of these 2 specimens had an S/CO >2.5, while the other had an S/CO ≤2.5.

CONCLUSION: A highly significant (<0.0001) correlation was found between the S/CO ratio on HCV EIA 1.0‐ and RIBA 2.0‐positive or ‐negative results. An S/CO ratio >2.5 yielded an 89‐ percent sensitivity for RIBA 2.0‐positive specimens, and donations with an S/CO ratio >2.5 had a 75‐percent probability of being RIBA 2.0 positive. A policy recommendation to use the S/CO ratio to triage lookback would prevent unwarranted notification of 87 percent of recipients of blood from RIBA 2.0‐negative donors and would result in a failure to notify only 5 to 10 percent of recipients potentially exposed to infectious units.
Preparation and storage characteristics of white cell‐reducedhigh‐concentration platelet concentrates collected by anapheresis system for transfusions in utero
Authors: Dumont L.J. , Krailadsiri P. , Seghatchian J. , Taylor L.A. , Howell C.A. , Murphy M.F.
Published: March 2003

BACKGROUND: Important concerns with regard to in utero platelet transfusions are avoidance of volume overload and the immunomodulatory effects of residual white cells (WBCs). This study evaluated a modification of a leukocyte‐reduction system (LRS, Spectra, COBE BCT) for apheresis, which collects high‐concentration WBC‐reduced platelets (HCPs) for in utero transfusion.

STUDY DESIGN AND METHODS: The LRS procedure was modified by running the platelet collection pump at specified low flow rates (Qcol) for the first part of the procedure, collecting HCPs by gently purging them from the LRS chamber into a designated collection bag and then restoring the original LRS procedure settings to collect a second standard apheresis platelet concentrate (PC). Two centers carried out 32 procedures. Platelet yield, residual WBCs, and in vitro platelet function studies were evaluated.

RESULTS: Platelet concentrations in 60 mL of HCPs were predictable according to Qcol (r2 = 0.735). HCP yields varied from 0.9 to 3.2 × 1011, depending on the desired final platelet concentrations in 60 mL, with an overall average of 1.92 × 1011 (n = 32). Apheresis PCs had a mean platelet yield of 2.9 × 1011 (1.3‐4.4 × 1011, n = 20) and 3.9 × 1011 (2.2‐5.8 × 1011, n = 12) at concentrations of 1.3 × 1012 per L for single‐needle and dual‐ needle procedures, respectively. Median WBC counts were 5.6 × 103 for HCPs and 2.0 × 104 for apheresis PCs, with >99 percent expected to be less than 1 × 106. HCP in vitro characteristics were equivalent to those of apheresis PCs at 24 hours after collection. In vitro performance declined over storage as a function of HCP yield. HCP pH at 22oC was maintained at a level of >6.2 for more than 3 days for yields >1.6 × 1011, less than 2 days for yields 1.6 to 2.2 × 1011, and less than 24 hours for yields >2.2 × 1011. HCPs showed good in vitro characteristics and could be stored for 1 to 3 days, depending on the total number of platelets collected.

CONCLUSION: A standard apheresis PC and an HCP requiring no secondary processing can be collected with the Spectra LRS. The platelet concentration may be determined by clinical need. HCPs meet the requirements for components that are transfused in utero.
Apheresis‐induced platelet activation:comparison of three types of cell separators
Authors: Hagberg I.A. , Akkøk Ç.A. , Lyberg T. , Kjeldsen‐Kragh J.
Published: March 2003

BACKGROUND: Platelet‐harvesting technology differs in various cell separators. Alteration in shear stress and biocompatibility of surfaces may give variable platelet activation and thereby affect the quality of the component.

STUDY DESIGN AND METHODS: Four groups (n = 10) of single‐needle apheresis procedures using three cell separators, were compared: 1) Spectra LRS, 90‐minute harvesting time; 2) MCS+, 90‐minute harvest; 3) Amicus, 90‐minute; and 4) Amicus, 45‐minute. Whole‐blood samples were collected from the donors as were samples from the final components at intervals during the first 4 hours after cessation of the apheresis. Platelet activation status and platelet activation capacity after agonist stimulation were assessed by flow cytometry.

RESULTS: No activated platelets were found in preapheresis and postapheresis samples from the donors. The platelets in the components from the Amicus (90‐min) were significantly more activated than those in the other groups of components: that is, there was increased size of platelet aggregates, increased fraction of microparticles, increased degranulation, increased fibrinogen receptor activation, and decreased von Willebrand factor receptor expression. Moreover, the response of these platelets to agonist stimulation was reduced for all activation variables.

CONCLUSIONS: After 90 minutes' processing time, platelets obtained with the Amicus cell separator were significantly more activated than platelets harvested with the Spectra and the MCS+.
A comparison of prestorage WBC‐reduced whole‐blood‐derived platelets and bedside‐filtered whole‐blood‐derived platelets in autologous progenitor cell transplant
Authors: Sweeney J.D. , Kouttab N.M. , Penn C.L. , McHugh K.E. , Nelson E.J. , Oblon D.J.
Published: March 2003

BACKGROUND : Prestorage WBC‐reduced platelet concentrates (PCs) can be manufactured from platelet‐rich plasma (PRP) by in‐line filtration of PRP. There are few published data on the clinical use of these products, as compared to bedside‐filtered pools of standard PCs (S‐PCs) manufactured from PRP.

STUDY DESIGN AND METHODS : A prospective, randomized trial was conducted in autologous progenitor cell transplant patients requiring platelet transfusions with each patient as his or her own control who was given a pool of 5 units of WBC‐reduced PCs and a pool of 6 units of S‐PCs within a 3‐hour period. The pools were characterized before transfusion for platelet and WBC content, P‐selectin expression, and IL‐8. The patients were monitored with platelet counts and vital signs and observed for reactions. Data were analyzed using Mann‐Whitney U tests.

RESULTS : Thirty‐three transfusions were administered to 13 patients. Median platelet content in the WBC‐reduced PC pools was lower than that in the S‐PC pools (3.3 vs. 4.0 × 1011, p<0.01). Median WBC content was 4 to 5 log less in the WBC‐reduced PC pools (2.5 × 104 vs. 4.6 × 108, p<0.01). Median IL‐8 levels (pg/mL) were lower in the WBC‐reduced PC pools (2 vs. 36, p<0.01). No differences were observed in CCI, but the median absolute increase after transfusion of the S‐PC pools was higher (25 vs. 19 × 109/L, p<0.01), which reflected the larger size of the S‐PC pools. No overall differences in vital signs were recorded. Two reactions were observed, both in temporal association with the transfusion of pools of S‐PCs.

CONCLUSIONS : A pool consisting of 5 units of WBC‐reduced PCs gave a median platelet increment of 19 × 109 per L in these thrombocytopenic patients and has a median WBC content 1 to 2 log below the accepted threshold for primary alloimmunization or CMV transmission.
The cost‐effectiveness of postoperative recovery of RBCs in preventing transfusion‐associated virus transmission after joint arthroplasty
Authors: Jackson Brian R. , Umlas Joel , AuBuchon James P.
Published: March 2003

BACKGROUND: The return of joint drainage after hip and knee arthroplasty is a widely used but expensive blood‐conservation technique.

STUDY DESIGN AND METHODS: A Markov decision analysis model was used to evaluate the cost‐effectiveness of postoperative RBC recovery in preventing viral complications of allogeneic transfusion.

RESULTS: In the baseline analysis, using an RBC‐recovery device saves 5 quality‐adjusted minutes of longevity at an average incremental cost of $53. This corresponds to $5.7 million per quality‐adjusted life year. This figure was most sensitive to the direct cost differences of allogeneic versus recovered RBCs and to the volume of RBCs recovered per device. Such devices would save health care resources if they cost less than $73 or if they were applied only in cases where joint drainage was between 600 and 1100 mL.

CONCLUSION: For most clinical situations, postarthroplasty RBC recovery does not appear to be as cost‐effective as most other medical interventions. Clinical attention should be directed toward developing protocols for the preferential use of postoperative RBC‐recovery devices in situations where they provide the greatest benefit.
Storage of G–CSF‐mobilized granulocyte concentrates
Authors: Lightfoot Thomas , Leitman Susan F. , Stroncek David F.
Published: March 2003

BACKGROUND: Current standards limit granulocyte storage to 24 hours. Since G–CSF inhibits granulocyte apoptosis, it may be possible to store G–CSF‐mobilized granulocytes for longer periods while maintaining cell viability and function. However, G–CSF mobilization increases the yield of granulocytes several times, and the resulting higher cell concentrations may diminish viability during storage and significant levels of pyrogenic cytokines may be produced.

STUDY DESIGN: Ten granulocyte donors were given dexamethasone (8 mg PO), G–CSF (5 μg/kg SQ), or both and on the next day granulocyte concentrates were collected using a blood cell separator. Component cell counts, cell viablilities, pH, and IL‐1β, IL‐6, IL‐8 and TNF levels were measured at 2 to 4 (2), 20 to 28 (24), and 44 to 52 hours (48 hours).

RESULTS: Significantly more granulocytes were collected when donors were given G–CSF (4.2 ± 2.3 × 1010) or G–CSF plus dexamethasone (6.4 ± 2.5 × 1010) compared with that collected with dexamethasone alone (2.2 ± 1.2 × 1010); p = 0.03 and p = 0.002, respectively. Storage had little effect on WBC count. Slight but significant increases in IL‐1β and IL‐8 occurred after 24 and 48 hours as compared to the levels at 2 hours' storage. Levels of IL‐6 and TNF did not change. The pH dropped significantly with time in granulocytes mobilized with each regimen. Granulocytes mobilized with G–CSF plus dexamethasone were acidic immediately after collection, and pH was below 6.0 after 24 hours. To assess the effect of cell concentrations on pH, serial dilutions were performed on 13 granulocyte concentrates in autologous plasma prior to storage. The pH remained above 7.0 only when dexamethasone‐mobilized granulocytes were diluted 1‐in‐8 and when the G–CSF plus dexamethasone‐mobilized granulocytes were diluted 1‐in‐16.

CONCLUSIONS: To optimize storage pH, mobilized granulocyte concentrates require a 1‐in‐8 to 1‐in‐16 dilution, which is operationally impractical. Clinical‐grade granulocyte preservative solutions are needed to maintain pH during storage.
Apparent rejuvenation of transfused donor blood in the fetus is due to accelerated removal of the older RBCs
Authors: Huisman Margriet A. , Egberts Johannes
Published: March 2003

BACKGROUND: In severely anemic fetuses of women alloimmunized to RBC antigens, transfused donor RBCs disappear faster than in adults. This may result from an accelerated linear or nonlinear decline with time. It was investigated whether changes in donor RBC age characteristics after circulation in the fetus may reflect the main type of cellular decline.

STUDY DESIGN AND METHODS: Donor RBC age characteristics (density, mean cell volume [MCV], and mean cell Hb content [MCHC]) were determined before intrauterine transfusions. Density gradient centrifugation was used to obtain RBCs of different ages. The results from gradient centrifugation were used to calculate mean values for the density, MCV, and MCHC to be expected after the transfusion interval, assuming a linear decline in RBCs of 1 percent per day. Donor and fetal RBCs, taken just before the second transfusion, were separated by agglutination with IgM D MoAb. For these donor cells, the observed mean values for density, MCV, and MCHC were compared with the calculated, expected values (n = 12).

RESULTS: The mean ± SD transfusion interval was 17.9 ± 3.6 days. The Hb declined by 1.75 ± 0.62 percent per day (n = 9). After the transfusion interval and contrary to the expected changes, cell density and MCHC decreased and MCV increased significantly (0.001<p<0.02). This difference between actual and calculated values decreased with increasing intervals; for MCV, it was also associated with a greater decline in Hb per day (p<0.05).

CONCLUSION: All donor cells age during circulation in the fetus. However, after the transfusion interval, the donor RBC population remaining is apparently younger than the RBC population before transfusion. This results from a preferential disappearance of older donor RBCs and not from a linear loss of cells with time. The removal of older RBCs before the transfusion may increase the time between transfusions and thereby reduce the total number of transfusions required.
An in vitro method for predicting the efficacy of WBC separation using different starch preparations and anticoagulant ratios
Authors: Rock Gail , Berger Rachel , Romans Robert A. , Russell Norma M. , Owens Wendy A.
Published: March 2003

BACKGROUND: Separation of blood components depends on a number of factors, including the viscosity of the plasma and the number and size of the various cellular elements. To enhance granulocyte collection, it is common practice to alter the plasma environment by the addition of sedimenting agents such as hydroxyethyl starch. Recently, because of its prolonged persistence in the circulation, the higher‐molecular‐weight form of hydroxyethyl starch, Hespan (HP), has been replaced by the lower‐molecular‐weight form, pentastarch (PS). However, the yield appears to be lower. A rapid in vitro approach was used to permit comparison of the efficiency of separation of WBCs by the use of PS and HP and different ratios of anticoagulants that also alter the sedimenting characteristics of blood.

STUDY DESIGN AND METHOD: Blood from individual persons was collected into sodium citrate at ratios of 1:8, 1:12, and 1:16. Samples were evaluated either before or after the addition of PS or HP and after centrifugation.

RESULTS: The addition of HP increased the sedimentation rate to at least four times that of plasma (10.9 vs. 47.9 mm); PS approximately doubled the rate. Viscosity was altered by the introduction of either starch. These changes (ranging from a rate of 4.2 in HP with a 1:16 anticoagulant to 3.6 in PS with a 1:8 ratio of anticoagulant) reflected the anticipated effects of anticoagulant dilution and carbohydrate addition. Granulocyte recovery was highest, with a 1:12 anticoagulant ratio in all tests with HP producing the greatest yield (HP, 101%; PS, 89%; control, 78%).

CONCLUSION: HP is far more effective than its lower‐molecular‐weight substitute PS in the generation of granulocytes in the buffy coat of whole blood. This method provides a simple, rapid, in vitro approach to evaluating the separating efficiency of solutions.
Isolation and flow cytometric analysis of T‐cell‐depleted CD34+ PBPCs
Authors: Debelak Joan , Shlomchik Mark J. , Snyder Edward L. , Cooper Dennis , Seropian Stuart , McGuirk Joel , Smith Brian , Krause Diane S.
Published: March 2003

BACKGROUND: To extend allogeneic HPC transplantation to a greater range of patients, the use of partially matched related donors is under development. Because of the inherently higher degree of histoincom‐patibility in such transplants, there is increased risk of GVHD as well as of graft failure. Ex vivo depletion of donor‐derived T‐lymphocytes from PBPCs is one of the most effective methods of preventing GVHD. Thus far, individual centers have used custom‐developed procedures to deplete the graft of T cells that are responsible for alloreactivity, often employing relatively impure, nonstandardized reagents such as soybean agglutinin and complement. In addition, with improved methods of T‐cell depletion, it has been difficult to accurately assess the number of T cells remaining. Because different centers have used different protocols to assay T cells, it has been difficult to reproduce and validate the results between institutions, and this has limited direct comparison of data between centers.

STUDY DESIGN AND METHODS: A standardized approach for T‐cell depletion was developed by using a Good Manufacturing Practice‐manufactured magnetic cell separator (Isolex 300i, Nexell Therapeutics) and commercially available OKT3 antibody. T‐cell depletion was performed on PBPCs from six haploidentical donors.

RESULTS: CD34+ cell recovery was 47 percent (range, 31‐63%) with a median purity of 94 percent (range, 75‐99%) and median T‐cell log depletion of 4.72 (range, 3.90‐5.83). Because this high degree of depletion makes it challenging to accurately quantitate the remaining T cells, two highly sensitive flow cytometric protocols were developed, each of which accurately detects T cells with a sensitivity of 2 per 10,000 (0.02%). The purified CD34+ cells administered to the patients (dose range, 6.13‐13.50 × 106/kg) provided rapid neutrophil and platelet engraftment.

CONCLUSION: With the Isolex 300i and a MoAb directed against T cells, a high degree of T‐cell depletion is obtained. Sensitive, accurate, and reproducible assays have now been developed for T‐cell enumeration in these highly purified cell populations.
Release of WBC‐derived IL‐1 receptor antagonist into supernatants of RBCs: influence of storage time and filtration
Authors: Büttnerova Ivana , Bäumler Hans , Kern Florian , Radtke Hartmut , Volk Hans‐Dieter , Kiesewetter Holger , Döcke Wolf‐Dietrich
Published: March 2003

BACKGROUND: Transfusion‐associated immunodepression may be related to the transfer of immunoinhibitory cytokines with blood components.

STUDY DESIGN AND METHODS: After evidence of increasing concentrations of IL‐1 receptor antagonist (IL‐1RA) but not of IL‐10 was obtained in supernatants of stored RBC units that were WBC‐reduced by centrifugation (C‐RBCs) in a pilot study, IL‐1RA concentrations were determined weekly in supernatants of C‐RBCs and in units that underwent prestorage WBC reduction by in‐line filtration (F‐RBCs) over a 49‐day storage. For assessing total IL‐1RA content, complete cell lysis by repeated freezing and thawing was done. The results were related to the changes in WBC count during storage. The dependency of IL‐1RA content on preparation procedures was assessed.

RESULTS: The prestorage IL‐1RA concentration in C‐RBCs (859 ± 218 pg/mL) was significantly higher than in F‐RBC (75 ± 13 pg/mL). Whereas no changes were seen in F‐RBCs during storage, IL‐1RA levels in C‐RBC supernatants drastically increased to levels about 50 times those in normal plasma (16,327 ± 2,686 pg/mL on Day 49). Follow‐up analysis revealed stringent correlation between IL‐1RA release into supernatants and the current loss of WBCs (r = 0.79, n = 42; p<0.001). The total IL‐1RA content did not change during storage and was directly dependent on prestorage WBC count. Preparation procedures altered the IL‐1RA content only by WBC reduction.

CONCLUSION: The immunosuppressive cytokine IL‐1RA is transmitted by RBCs in relation to WBC content and storage time.
Statistical process monitoring of WBC‐reduced blood components assessed by two types of software
Authors: Seghatchian Jerard , Krailadsiri Pranee , McCall Michael
Published: March 2003

BACKGROUND: Statistical process control is required for monitoring of the WBC‐reduction process. This study focused on some factors that may influence the outcomes of statistical process monitoring, such as WBC‐reduction technologies, the anticoagulant used, and WBC‐counting technologies, by using two types of software.

STUDY DESIGN AND METHODS: Data were collected from January to September 1999, before the implementation of universal WBC reduction. The effects of three major factors were investigated: methods of preparation, the addition of EDTA to the sample, and the WBC‐counting technologies used (microvolume fluorimetry, flow cytometry, and Nageotte chamber). The WBC‐reduction process capability was assessed by two types of software, EZQC (Gambro BCT) and NWA (Northwest Analytical). In addition, the differences between various sets of results were compared by the t test or ANOVA.

RESULTS: There was no statistical difference (at the 0.05 level of significance) in WBC content when the three types of platelets in citrate samples were compared with EDTA samples. In general, the Nageotte chamber appeared to count the lowest, and microvolume fluorimetry appeared to count lower than flow cytometry. There were minor but significant methodologic differences between the software packages. However, these differences had negligible effects on the percentage of conforming components at both <1 × 106 and <5 × 106 WBCs per unit.

CONCLUSION: Only the counting technologies were sufficiently different to warrant consideration. This difference may make unacceptable the interchange of results obtained from various counting methods.
Seroprevalence of known and putative hepatitis markers inUnited States blood donors with ALT levels at least 120 IU per L
Authors: Notari Edward P. , Orton Sharyn L. , Cable Ritchard G. , Grindon Alfred J. , Lenes Bruce A. , Williams Alan E. , McMillan Kimberly M. , Trouern‐Trend Jonathan J. , Wolf‐Nugent Jennifer S. , Xu Yi‐Ling , Dodd Roger Y. ,   for the ARCNET Program
Published: March 2003

BACKGROUND: ALT testing of blood donors was initiated as a surrogate marker for non‐A, non‐B hepatitis. Increased sensitivity of subsequent HBV and HCV tests used for standard donor screening make any residual value of ALT testing questionable.

STUDY DESIGN AND METHODS: A prospective study was conducted in 166 of 645 eligible blood donors from three American Red Cross regions whose ALT was ≥120 IU per L and whose standard donor screening tests were negative. Of these enrolled donors, 124 (75%) completed follow‐up. Samples obtained from the index donation, at enrollment (1 month), and at follow‐up (6 months) underwent the standard donor screening tests, as well as those for HCV RNA and HGV RNA (RT‐PCR), antibodies to the virus envelope E2 protein of GB virus type C (GBV‐C E2 antibody), and IgM antibody for CMV, parvovirus B19, EBV VCA, and HAV. Participants completed a brief demographic and exposure history questionnaire at follow‐up.

RESULTS: All study samples were negative in standard donor‐screening tests. ALT levels were variable at return visits, with 80 to 86 percent <120 IU per L. No participants were positive for HCV RNA; 4 percent were positive for HGV RNA, and 10 percent were positive for GBV‐C E2 antibody. Results of CMV, parvovirus B19, EBV VCA, and HAV testing were similar to published background rates. No demographic or exposure history variables had significant correlation with ALT or other testing results.

CONCLUSION: These data suggest that an ALT ≥120 IU per L in blood donors with negative standard screening tests has questionable value as a surrogate marker for seronegative HBV or HCV infection. Continued ALT testing may contribute little, if anything, to the safety of blood components or plasma for further manufacture.