bluuebunny/crossref_metadata_2025 huggingface dataset:
Processing of major ABO‐incompatible bone marrow for transplantation by using dextran sedimentation
Authors: Tsang K.S. , Li C.K. , Wong A.P. , Leung Y. , Lau T.T. , Li K. , Shing M.M.K. , Chik K.W. , Yuen P.M.P.
Published: March 2003

BACKGROUND: Various open and semi‐closed methods are used for red cell (RBC) depletion and hematopoietic progenitor cell (HPC) enrichment of bone marrow (BM) in vitro, but with variable efficacy. A simple, efficient, and safe method using dextran 110k was developed.

STUDY DESIGN AND METHODS: An equal volume of 4.5‐percent dextran was applied to major ABO‐incompatible BM in transfer bags and sedimentation was allowed for 30 minutes. RBCs, nucleated cells (NCs), and mononuclear cells (MNCs) from BM allografts before and after dextran sedimentation (DS) were counted. Flow cytometry, short‐term cultures, and long‐term cultures were performed to assay the respective recovery of CD34+ cells, colony‐forming units (CFUs), and long‐term culture‐initiating cells (LTC‐ICs).

RESULTS: Sixteen BM collections were processed. The mean volume was 666 mL (range, 189‐1355 mL). The mean ± 1 SD post‐DS NC, MNC, CD34+ cell, and CFU counts per kg of the recipient's body weight were 4.11 ± 1.74 × 108, 8.98 ± 3.68 × 107, 2.90 ± 1.95 × 106, and 2.03 ± 2.01 × 105, respectively, with the corresponding post‐DS recovery being 90.6 percent, 90 percent, 92.4 percent, and 100.8 percent. The numbers of LTC‐ICs in cultures (up to 12 weeks) of pre‐DS and post‐DS samples of five BM allografts were comparable (p = 0.91). Residual RBCs were 5.1 ± 4.6 (0.1‐14) mL with depletion of 96.5 ± 3.2 percent. There was no significant difference in the mean absolute RBC count in post‐DS BM allografts and in four ficoll‐treated BM allografts (8.09 × 1010 vs. 4.9 × 109; p = 0.206) and in eight major ABO‐incompatible peripheral blood HPC collections (8.09 × 1010 vs. 9.81 × 1010; p = 0.87). No posttransplant hemolysis was encountered. Engraftment occurred at 22 ± 7 days, which is similar to that of four transplants with ficoll‐treated BM allografts (22 ± 9; p = 0.611) and 54 unprocessed BM allografts (19 ± 6; p = 0.129).

CONCLUSION: DS is an efficient method of depleting RBCs in major ABO‐incompatible BM allografts without significant loss of HPCs.
Transfusion with blood from a donor with chronic myelogenous leukemia: persistence of the bcr/abl translocation in the recipient
Authors: Vargas Sara O. , Cannon Marie E. , Benjamin Richard J. , Longtine Janina A.
Published: March 2003

BACKGROUND: Transfusion of cells harvested from patients with chronic myelogenous leukemia (CML) as a therapeutic measure for patients with granulocytopenia was popular in the 1970s, when studies examining the persistence of transfused donor cells were limited by a lack of molecular techniques. Blood samples from a patient who recently received an inadvertent transfusion of CML cells were evaluated for the presence of the bcr/abl translocation characteristic of CML.

CASE REPORT: The patient, a 67‐year‐old man with a history of congestive heart failure, myocardial infarct, hypertension, diabetes mellitus, and chronic renal failure, was transfused for bleeding from colonic angiodysplasia. A volunteer blood donor reported that he had been diagnosed with CML 10 days after his donation. Three days after the donation, blood components from the donor with CML had been administered to the patient as nonirradiated red cells and platelets. Evaluation of donor blood by a reverse‐transcriptase polymerase chain reaction showed the b3a2 transcript, indicating a bcr/abl translocation. Periodic testing of the patient's peripheral blood by the same technique demonstrated the presence of the b3a2 transcript on Days 74 and 75 after transfusion. The patient died of congestive heart failure 8 months after the transfusion.

CONCLUSION: In this rare case of accidental transfusion of neoplastic cells, the findings document the persistence of the donor's neoplastic clone in the recipient for 75 days.
High deferral rate for maternal–neonatal donor pairs for an allogeneic umbilical cord blood bank
Authors: Jefferies Leigh C. , Albertus Mary , Morgan Mark A. , Moolten David
Published: March 2003

BACKGROUND: To fully evaluate the safety and efficacy of umbilical cord blood (UCB) as a source of hematopoietic progenitor cells, UCB banks are being established worldwide to provide a readily accessible resource of stem and progenitor cells for allogeneic transplantation. Guidelines for UCB donor criteria have been proposed to minimize the possibility of disease transmission to UCB recipients.

STUDY DESIGN AND METHODS: In preparation for the establishment of an allogeneic UCB bank, 285 maternal–neonatal donor pairs were evaluated in our tertiary‐care hospital, which was chosen because of the high percentage of African Americans among the maternal population. Maternal, neonatal, and family histories were obtained by confidential interview and medical record review to determine the number of eligible donor pairs on the basis of a conservative interpretation of proposed donor criteria.

RESULTS: Only 44 percent of donor pairs were considered eligible. The most common deferral factors were maternal, and they included fever at delivery, history of chronic disease, and history of sexually transmitted disease. In most cases, more than one deferral factor was identified.

CONCLUSIONS: High‐risk maternal populations, which may provide access to ethnic minorities targeted for some UCB banks, may contain low percentages of eligible donors. Further refinement of donor criteria will be important in the evolution of UCB banking and transplantation.
Safety issues of plateletpheresis: comparison of the effects of two cell separators on the activation of coagulation, fibrinolysis, and neutrophils and on the formation of neutrophil‐platelet aggregates
Authors: Stohlawetz Petra , Kapiotis Stylianos , Seidl Dagmar , Hergovich Nicole , Zellner Maria , Eichler Hans‐Georg , Stiegler Gabriele , Leitner Gerda , Höcker Paul , Jilma Bernd
Published: March 2003

BACKGROUND: Although many donors undergo repeated plateletpheresis, data on the consequences of plateletpheresis for the donor's health remain scarce. Thus, the effect of plateletpheresis on the activation of coagulation, fibrinolysis, and neutrophils was investigated.

STUDY DESIGN AND METHODS: Part 1: Sixteen healthy men were randomly assigned to undergo plateletpheresis on a cell separator (AMICUS, Fenwal Baxter; or MCS 3p, Haemonetics). The effects of plateletpheresis on plasma levels of prothrombin fragment (F1+2), d‐dimer, plasmin‐plasmin inhibitor (PPI) complexes, and plasminogen activator inhibitor (PAI‐1); on the activation of neutrophils (% l‐selectin+); and on the frequency of platelet‐neutrophil aggregates (% CD41 + neutrophils) were compared. Part 2: Ten healthy men received infusions of ACD‐A and placebo without apheresis in a randomized, double‐blind crossover study to control for the pharmacologic effects of citrate.

RESULTS: Part 1: No change in F1+2 occurred (p>0.05), which indicated that plateletpheresis did not enhance coagulation. Levels of d‐dimer, PPI, and PAI‐1 decreased over time on the AMICUS (p<0.001). Plateletpheresis did not activate neutrophils (p>0.05), but it decreased the percentage of CD41+ neutrophils (p<0.003). An approximately 80‐percent drop in mononuclear cells was observed in the extracorporeal circulation of the AMICUS (p<0.001 vs. baseline and p = 0.005 vs. MCS 3p), and circulating lymphocyte and monocyte counts decreased concomitantly. Part 2: Infusion of ACD‐A slightly decreased d‐dimer levels (p<0.05), and both infusions decreased the circulating lymphocyte counts.

CONCLUSION: Plateletpheresis can be regarded as safe with respect to the activation of coagulation or neutrophils. The consequences for the donor's health of the decrease in d‐dimer, PPI, and PAI‐1 may deserve further investigation.
GVHD after transfusion of stored RBC concentrates in asolution of mannitol, adenine, phosphate, citrate, glucose,and NaCl following trauma
Authors: Kita T. , Nei J. , Matsu T. , Yoshiba H. , Matsuda T. , Yahagi H. , Uchida S. , Nakao S.
Published: March 2003

BACKGROUND: It has not previously been reported that WBC‐reduced RBC preparations can cause transfusion‐associated GVHD, even in an immunocompetent individual.

CASE REPORT: A 74‐year‐old man suffered a hemorrhage from the mesentery of the transverse colon after a traffic accident. During surgery, he received 10 units of RBCs from 10 donors in a solution containing mannitol, adenine, phosphate, citrate, glucose and NaCl (MAP). MAP RBCs had been stored for 7 to 8 days before use. On the 27th day after surgery, an erythematous, pruritic rash appeared over the face, neck, and trunk, which was associated with low‐grade fever and pancytopenia. Transfusion‐associated GVHD was strongly suspected and was confirmed by skin biopsy. To determine the origin of lymphocytes causing GVHD, several microsatellite loci were amplified from DNA of the patient's nails and blood and from blood samples of all 10 RBC donors by using PCR. Amplified alleles derived from the patient's blood were identical to those from one of the 10 samples.

CONCLUSION: These findings indicate that transfusions of MAP‐RBCs can cause transfusion‐associated GVHD in an elderly but immunocompetent host.
Enhanced HPC recruitment in children using LVL and a new automated apheresis system
Authors: Torrabadella M. , Olivé T. , Ortega J.J. , Massuet L.
Published: March 2003

BACKGROUND: A new automated apheresis system has recently been reported as useful in improving peripheral blood HPC collection in adults. The aim of this study has been to verify the utility of this system (AutoPBSC, COBE BCT) for standard leukapheresis and for LVL in the pediatric setting.

STUDY DESIGN AND METHODS: A prospective study was set up in 29 leukapheresis procedures carried out in 26 children with malignant diseases and body weight under 40 kg who had undergone mobilization with G–CSF or with G–CSF and chemotherapy. Leukapheresis procedures were performed under two protocols, depending on the total blood volume processed: standard leukapheresis (≤3) and LVL (>3). The need to prime the tubing set with blood was determined, and the inlet flow rate, collection time, recruitment of CD34+ cells, CD34+ cell collection efficiency, component volume, leukapher‐esis cell composition, and preapheresis and postapher‐esis peripheral blood counts were measured. Paired t test, Spearman's correlation coefficient, and the Mann‐Whitney U test were employed for statistical analysis.

RESULTS: Because of the low extracorporeal volume (167 mL) of the tubing set of the automated blood processor, priming was necessary in only 2 of 26 patients, both weighing under 10 kg. LVL showed better CD34+ cell yield (7.5 vs. 2.3 × 106/kg; p = 0.047), higher recruitment (2.1 vs. 0.9; p = 0.002), and greater collection efficiency (50% vs. 33%; p = 0.005) than standard leukapheresis. No significant differences were found between groups in collection time. In LVL procedures, CD34+ cell collection efficiency and recruitment were not significantly influenced by the inlet flow rate.

CONCLUSION: The AutoPBSC is a reliable system for peripheral blood HPC collection in children mainly when used in combination with LVL. The major advantage of this software is a reduced need for priming. LVL allows better CD34+ cell collection efficiency, enhanced recruitment, and improved CD34+ cell yield.
Fatigue during acute isovolemic anemiain healthy, resting humans
Authors: Toy P. , Feiner J. , Viele M.K. , Watson J. , Yeap H. , Weiskopf R.B.
Published: March 2003

BACKGROUND: Transfusion guidelines recommend that clinicians assess patients for signs and symptoms of anemia before the transfusion of RBCs. However, studies of signs and symptoms associated with acute isovolemic anemia are limited. The objective of this study was to determine whether acute reduction of Hb concentration to 5 g per dL would result in fatigue, tachycardia, or hypotension in resting, young, healthy, isovolemic humans, and whether changes were reversible with RBC transfusion.

STUDY DESIGN AND METHODS: Conscious, resting, healthy adults less than 35 years old (n = 8) underwent acute isovolemic hemodilution to Hb of 5 g per dL and self‐scored their energy level at various Hb concentrations. Heart rate and blood pressure were also measured. For controls, measurements of each subject were made during a comparable period of rest without hemodilution.

RESULTS: During acute isovolemic hemodilution, energy levels decreased progressively and were lower at Hb of 7, 6, and 5 g per dL than at baseline (p<0.01) or in control sessions (p<0.05). The energy level was lower at Hb 7 g per dL than at 14 ( p = 0.005), lower at Hb 6 g per dL than at 7 (p = 0.0l), and lower at Hb 5 g per dL than at 6 (p =0.01). Energy levels rose and were not different from baseline or control levels after transfusion of all autologous RBCs. Similarly, median heart rate increased with hemodilution to Hb of 7, 6, and 5 g per dL and decreased with transfusion of autologous RBCs. Supine blood pressure did not decrease with isovolemic hemodilution.

CONCLUSION: In resting, young, healthy humans, acute isovolemic anemia to Hb levels of 7, 6, and 5 g per dL results in decreased self‐scored energy levels and in an increase in heart rate but not in hypotension. Changes in energy and heart rate are reversible with the transfusion of autologous RBCs.
Evaluation of automated RNA‐extraction technology and a qualitative HCV assay for sensitivity and detection of HCV RNA in pool‐screening systems
Authors: Beld M. , Habibuw M.R. , Rebers S.P.H. , Boom R. , Reesink H.W.
Published: March 2003

BACKGROUND: The objective of this study was the evaluation of NAT technology for the detection of HCV RNA in plasma pools according to the recommendations of the Paul Ehrlich Institute (5000 IU/mL/donation) and the Committee for Proprietary Medical Products (100 IU/mL/manufacturing pool).

STUDY DESIGN AND METHODS: Serial dilutions of both the EUROHEP standard (3,800 genome equivalents [geq]/mL; HCV genotype 1) and the World Health Organization (WHO) international standard (100,000 IU/mL; HCV genotype 1) were made in S/D plasma (ESPEP plasma, OctaPharma), which was nonreactive in serologic tests. Serial dilutions of plasma (2 mL) were used for extraction of HCV RNA with an automated version of a nucleic acid isolation method (NucliSens Extractor, Organon Teknika). HCV RNA was co‐extracted from 2 mL of plasma, together with 84 copies of an in vitro‐synthesized single‐strand RNA serving as internal extraction control (IC) to monitor the efficiency of extraction and PCR. Amplification and detection of both HCV RNA and IC RNA were performed with an automated PCR system and a qualitative HCV assay (COBAS Amplicor 2.0 HCV, Roche Diagnostics).

RESULTS: A cutoff value of 16 geq per mL (10/10 runs [100% hit rate]) was found by using the EUROHEP standard, whereas the WHO international standard had a cutoff value of approximately 12 IU per mL (10/10 runs [100% hit rate]). The IC had a cutoff value of approximately 17.5 copies per mL (6/6 runs [100% hit rate]). Forty‐two copies per mL of IC RNA were found in 282 of 284 runs (99% hit rate). The negative controls (ESDEP plasma) were negative in all experiments. Experiments with pool sizes of 12, 24, 48, and 96 using serial dilutions of the WHO international standard revealed a cutoff value of 8 IU per mL (100% hit rate). The EUROHEP standard and the WHO international standard were detected with a 50 percent detection endpoint of 5.2 geq per mL and 1.5 IU per mL, respectively.

CONCLUSION: This test system (NucliSens Extractor, and the COBAS Amplicor 2.0 HCV assay) revealed a high sensitivity for HCV RNA; considering the proposed requirements for sensitivity of NAT assays for the detection of HCV RNA in donor plasma, pool sizes of about 400 donors are possible. These endpoint results indicated that 1 IU is equal to about 3.4 geq.
WBC subset analysis of WBC‐reduced platelet components
Authors: Triulzi D.J. , Meyer E.M. , Donnenberg A.D.
Published: March 2003

BACKGROUND: WBC‐reduced platelet components may be prepared by filtration or apheresis processing. Both methods have previously been shown to result in a residual total WBC content <5 × 106 per component. However, there may be differences in the efficacy of these techniques for removing certain WBC subsets.

STUDY DESIGN AND METHODS: Two multiparameter flow cytometric assays were developed and validated to perform WBC analysis on WBC‐reduced platelets collected with two apheresis instruments (Amicus and COBE Spectra) and on 6 units of filtered pooled random‐donor platelet concentrates.

RESULTS: All components contained <1 × 105 WBCs. The COBE Spectra and Amicus apheresis platelet components contained more WBCs than did filtered pooled platelets (p<0.05). Lymphocytes (T and B), monocytes, and granulocytes were identified in all components. Granulocyte content was lowest in the Amicus components and filtered pools. Monocytes were lowest in filtered pools. Amicus platelet components had fewer granulocytes and monocytes than the COBE Spectra platelets. Amicus and COBE Spectra components contained more lymphocytes than the filtered pools.

CONCLUSION: Multiparameter flow cytometry can be used to quantify and characterize WBCs in WBC‐reduced platelet components. WBC reduction by filtration or apheresis was highly effective. WBCs from each subset were identified in all components. Although filtered pools had the lowest numbers of WBCs, the very low numbers observed in all components suggests that the absolute quantitative differences in WBC subset content are of questionable clinical significance.
Complications of plasma exchange in 71 consecutive patients treated for clinically suspected thrombotic thrombocytopenic purpura‐hemolytic‐uremic syndrome
Authors: Rizvi M.A. , Vesely S.K. , George J.N. , Chandler L. , Duvall D. , Smith J.W. , Gilcher R.O.
Published: March 2003

BACKGROUND: With the increased frequency of diagnosis and improved survival of thrombotic thrombocytopenic purpura‐hemolytic‐uremic syndrome (TTP‐HUS), the morbidity of plasma exchange (PE) treatment has become more important.

STUDY DESIGN AND METHODS: Data were prospectively collected on 71 consecutive patients referred to the Oklahoma Blood Institute (OBI) for PE treatment for clinically suspected TTP‐HUS from mid‐1996 to mid‐1999. Complications were defined as major or minor, and distinguished between those related to central venous catheter access or to the plasma.

RESULTS: Twenty‐one patients (30%) had 27 major complications, which caused two deaths. The major complications included 2 episodes of hemorrhage after subclavian line insertion (1 death), 1 pneumothorax requiring a chest tube, 12 systemic infections (1 death), 7 episodes of catheter thrombosis requiring removal of the central venous catheter, 2 episodes of venous thrombosis requiring anticoagulant treatment, 2 episodes of hypoxemia and hypotension, and 1 episode of serum sickness. Minor complications occurred in 22 additional patients (31%). Twenty‐eight patients (39%) had no complications.

CONCLUSIONS: The morbidity and mortality of catheter placement and PE are important considerations when PE treatment for clinically suspected TTP‐HUS is anticipated.
Vaccination‐associated immune hemolytic anemia in two children
Authors: Seltsam A. , Shukry‐Schulz S. , Salama A.
Published: March 2003

BACKGROUND: Two children in whom acute autoimmune hemolytic anemia (AIHA) developed after vaccination were studied.

CASE REPORTS: The children were a 20‐month‐old girl and a 21‐month‐old boy. The diagnosis of AIHA was made in accordance with established criteria (hemolysis, positive DAT, and lack of other reasons for the hemolysis). Serologic tests were performed according to standard technique.

RESULTS: The girl experienced two attacks of hemolysis. The first episode occurred 2 weeks after oral polio vaccination, and the second episode was observed 7 months later, when she received a simultaneous vaccination against mumps, rubella, and measles. The DAT was strongly positive with anti‐C3d. No autoantibodies were detectable in either episode. The boy experienced acute hemolysis a few days after a simultaneous revaccination against diphtheria‐pertussis‐tetanus, Haemophilus influenzae, hepatitis B, and polio. The DAT using anti‐IgG was strongly positive, and the DAT performed with anti‐C3d was weakly positive.

CONCLUSION: Vaccination‐induced AIHA resembles those forms of AIHA related to infectious diseases, and it may occur more frequently than has been reported.
Thrombopoietin and the TPO receptorduring platelet storage
Authors: Xia Y. , Li J. , Bertino A. , Kuter D.J.
Published: March 2003

BACKGROUND: For most cells, the addition of a specific growth factor has improved cellular viability by preventing programmed cell death (apoptosis). To determine whether the platelet‐specific hematopoietic growth factor thrombopoietin (TPO) might improve platelet viability, endogenous TPO and the platelet TPO receptor were analyzed during storage, and the effect of recombinant TPO on platelet viability was assessed.

STUDY DESIGN AND METHODS: During platelet storage, TPO stability was assessed by SDS‐PAGE, TPO receptor function was measured, and the platelet TPO receptor was characterized by a 125I‐rHuTPO competitive‐binding assay. A recombinant TPO, pegylated recombinant human megakaryocyte growth and development factor (PEG‐rHuMGDF), was added to platelet concentrates during storage, and its effect on pH, LDH, and metabolic activity was determined.

RESULTS: During storage, the molecular weight and concentration of endogenous TPO (125 ± 19 pg/mL) and exogenous TPO (5720 ± 140 pg/mL) were constant for 12 days; the number (33 ± 4), binding affinity (149 ± 33 pM), and function of the platelet TPO receptors were constant for 7 days. Metabolic activity measured with the MTT and MTS assays closely correlated with changes in the pH and LDH. The addition of PEG‐rHuMGDF did not alter the pH, LDH, or metabolic activity of platelets during storage, but it did increase by 65 percent the uptake of 35S‐methionine into platelets. Finally, platelet concentrates obtained from donors treated with PEG‐rHuMGDF retained normal metabolic activity for 12 days, as compared with 5 to 6 days for normal platelet concentrates.

CONCLUSIONS: TPO and its platelet receptor are present in normal amounts and have normal function during platelet storage. The addition of recombinant TPO increased platelet methionine transport but did not alter platelet viability during storage. Other means to prevent apoptosis during platelet storage should be considered, and the measurement of platelet metabolic activity by MTT and MTS assays may assist this effort.
An evaluation of the process and costs associated with targeted lookbacks for HCV and general notification of transfusion recipients
Authors: Callum Jeannie L. , Pinkerton Peter H. , Coovadia Ahmed S. , Thomson Anne E. , Dewsbury Frankie
Published: March 2003

BACKGROUND: The Commission of Inquiry on the Blood System in Canada recommended that hospitals notify patients who received blood between 1978 and May 1990 of the risks of contracting HIV (up to the end of 1985 only) and HCV infection. The commission also recommended that patients should be informed of any transfusion received.

STUDY DESIGN AND METHOD: General notifications for HIV and HCV for this period were begun in mid‐1994. Notification after discharge of transfusions received after May 1990 was begun in 1997. Targeted HCV lookback was performed from 1995 to 1999.

RESULTS: Of 21,016 transfusion recipients from January 1978 to May 1990 identified in the general look‐back process and believed still alive, 13,549 (64%) were presumed contacted, by registered mail. The overall contact rate for the ongoing notifications (transfusions after May 1990) cannot be accurately determined, as registered mail was not used and a reply not requested. The total cost for these two processes was CAN$373,481, or $13 per patient believed contacted. Most (56%) of this cost was for the conversion to elec‐tronic form of paper transfusion records for the period 1978 through early 1984. In the targeted HCV lookback program 1995 through 1999, 94 percent of 256 recipients of specific components identified as likely to have transmitted HCV either were contacted or had died. Of 84 living recipients, 47 (56%) are HCV positive. The last documented potential serocon‐version occurred after a transfusion in November 1991, during the period of first‐generation EIA testing. If the targeted HCV lookback had been restricted to transfusions after 1987, as the FDA recommended, we would have failed to identify 39 living patients, of whom 21 are HCV positive. The cost per HCV‐positive patient notified in the targeted HCV lookback was CAN $4,174.

CONCLUSION: The cost of compliance with the com‐mission's recommendations was CAN$569,636. Over 28,000 of 36,773 transfusion recipients were notified or presumed notified, and 272 targeted HCV lookbacks to 256 recipients were performed. Performance of this task required the existence of transfusion records back to 1978, conversion of paper records to electronic form, and adequate secretarial and financial support.
A model of the health and economic impact of posttransfusion hepatitis C: application to cost‐effectiveness analysis of further expansion of HCV screening protocols
Authors: Pereira Arturo , Sanz Cristina
Published: March 2003

BACKGROUND: Cost‐effectiveness analyses are needed to decide the value of further expansion of the screening protocols for HCV in blood donors. However, such analyses are hampered by imperfect knowledge of the health and economic repercussions of posttransfu‐sion hepatitis C (PTHC).

STUDY DESIGN AND METHODS: A Monte Carlo simulation of a Markov model representing the outcomes of patients transfused with HCV‐infective blood was used to estimate the health and economic impact of PTHC and to calculate the cost‐effectiveness ratio of various HCV screening methods.

RESULTS: Median survival for hypothetical patients with PTHC and for controls without hepatitis was 11.25 and 11.75 years, respectively. Overall, 12.3 percent of patients receiving HCV‐infective blood will develop chronic hepatitis, 9.3 percent will progress to liver failure, and 9.25 percent will eventually die of liver disease after a median time of 20.75 years (range, 6‐70). Ninety‐one percent of the infected blood recipients had no reduction in life expectancy due to PTHC, and the average loss per patient was 0.754 years. The present value of the lifetime health costs incurred by patients with PTHC is $6330 per case. HCV antibody testing increases the patients' life expectancy by 20.4 hours per blood collection tested, and it results in net savings by decreasing the number of patients that will require treatment for liver disease in the future. Adding HCV NAT increases the patients' life expectancy by 0.08 hours per blood collection tested, at a cost‐effectiveness ratio of $1,829,611 per QALY gained.

CONCLUSION: PTHC has low health benefits because of the advanced age of many blood recipients. Testing donors for HCV antibodies results in net savings for the health care system, despite low health benefits. Adding HCV NAT would produce little additional gain at a very high cost.
Immunomagnetic tumor cell selection—implications for the detection of disseminated cancer cells
Authors: Krüger William , Datta Chonda , Badbaran Anita , Tögel Florian , Gutensohn Kai , Carrero Isabel , Kröger Nicolaus , Jänicke Fritz , Zander Axel R.
Published: March 2003

BACKGROUND: The optimal method for the detection of disseminated epithelial cancer cells has not yet been found. The standard method, using immunocytochemistry, offers a sensitivity of up to 10−6. Molecular methods such as cytokeratin‐19 RT‐PCR are about 10 times as sensitive, but they are hampered by interference such as illegitimate gene expression.

STUDY DESIGN AND METHODS: Immunomagnetic bead selection of epithelial cancer cells using conjugates directed against the human epithelial antigen (HEA) followed by immunocytochemistry testing was investigated in this trial.

RESULTS: No cytokeratin‐positive cells could be enriched from 56 control samples. In 104 clinical samples of bone marrow aspirations, PBPC collections, and venous blood obtained from breast cancer patients, the cytokeratin‐positive rate increased significantly, from 29.9 percent before selection to 54.8 percent after enrichment. Even the yield of detected cancer cells was significantly higher after selection. Up to 2.5 × 108 MNCs were easily processed. However, the mean cancer cell recovery after HEA enrichment was only 24.4 percent. Subsequently, selected epithelial cells were successfully immunophenotyped by use of a double‐stain technique detecting cytokeratin‐positive cells and the urokinase‐like plasminogen activator receptor.

CONCLUSION: HEA bead selection in combination with the standard immunocytochemistry method is a powerful and specific tool for the detection of disseminated cancer cells without false‐positive results. Furthermore, it delivers enough cells for subsequent investigations such as characterization studies.
Receptors involved in the phagocytosis of senescent and diamide‐oxidized human RBCs
Authors: Tartakover‐Matalon Shelly , Shoham‐Kessary Hadar , Foltyn Vera , Gershon Harriet
Published: March 2003

BACKGROUND: Senescent RBCs bear IgG and C3 opsonins that are three to four times less than required for similar phagocytosis of experimentally opsonized RBCs.

STUDY DESIGN AND METHODS: Studies were performed to determine the phagocyte receptors involved in phagocytosis in vitro. The effect of clustering of opsonins and oxidative damage in the sequestration of RBCs was studied by exposing RBCs to BS3 (bis[sulfosuccinimidyl]‐suberate) and diamide (azodicarboxylic acid bis[dimethyl‐amide]).

RESULTS: Sequestration of senescent RBCs was inhibited by the treatment of lymphokine‐activated monocytes with N‐acetyl‐D‐galactoseamine (GalNAc), arginine‐glycine‐aspartic acid (RGD), or antibodies to CR3, FcγRI, FcγRII, leukocyte response integrin (LRI), and integrin‐associated protein (IAP). Exposure to BS3 alone did not enhance phagocytosis. The addition of serum resulted in opsonin binding. The level of opsonization required for sequestration was higher than on senescent RBCs and was only marginally inhibited by blocking CR3, FcγRI and FcγRII. Diamide treatment alone did not lead to sequestration. Diamide‐treated RBCs exposed to serum bound opsonin much as did senescent RBCs, and sequestration was inhibited by GalNAc, RGD, and antibodies to CR3, FcγRI, FcγRII, LRI, and IAP.

CONCLUSION: Membrane alterations resulting in the binding of opsonins and the sequestration of senescent RBCs may be similar to those that occur on diamide‐oxidized RBCs. They suggest the need for cooperative events among oxidation, clustering and cross‐linking, and serum opsonization.
In vitro and in vivo measurements of human RBCs frozen with glycerol and subjected to various storage temperatures before deglycerolization and storage at 4°C for 3 days
Authors: Valeri C. Robert , Pivacek Linda E. , Cassidy George P. , Ragno Gina
Published: March 2003

BACKGROUND: This study was designed to assess the effects of changes in storage temperature of frozen RBCs such as might occur during a malfunction of the −80°C mechanical freezer or during shipment.

STUDY DESIGN AND METHODS: Fifteen participants donated blood for autologous transfusion of RBCs; all RBCs were frozen with 40‐percent (wt/vol) glycerol. Five subjects received RBCs that were stored at −80°C alone before transfusion. Five subjects received RBCs that were stored initially at −80°C, then at −40°C for 4 weeks, and finally at −80°C before transfusion. Five subjects received RBCs that were stored at −80°C, then at −20°C for 2 weeks, and finally at −80°C before transfusion. After deglycerolization, the RBCs were stored at 4°C in a sodium chloride‐glucose solution for 3 days before transfusion.

RESULTS: No significant differences were observed in freeze‐thaw recovery, freeze‐thaw‐wash recovery, 24‐hour posttransfusion survival, index of therapeutic effectiveness, or RBC ATP levels. Greater hemolysis and reduced RBC K+ levels were observed in the units stored at −80°C/−40°C/−80°C and in those stored at −80°C/ −20°C/−80°C compared with the units stored at −80°C alone, but these differences did not affect the 24‐hour posttransfusion survival.

CONCLUSIONS: The results of this study indicated that RBCs frozen with 40‐percent (wt/vol) glycerol can be stored at −40°C for 4 weeks or at −20°C for 2 weeks between periods of frozen storage at −80°C with satisfactory results.
Inactivation of parvovirus B19 in coagulation factor concentrates by UVC radiation: assessment by an in vitro infectivity assay using CFU–E derived from peripheral blood CD34+ cells
Authors: Sugawara Hiroyuki , Motokawa Reiko , Abe Hideki , Yamaguchi Miki , Yamada‐Ohnishi Yoshiko , Hirayama Junichi , Sakata Hidekatsu , Sato Shinichiro , Kamo Naoki , Ikebuchi Kenji , Ikeda Hisami
Published: March 2003

BACKGROUND: Nonenveloped and thermostable viruses such as parvovirus B19 (B19) can be transmitted to patients who are receiving plasma‐derived coagulation factor concentrates treated by the S/D method for inactivating enveloped viruses. Therefore, it is important to develop and validate new methods for the inactivation of nonenveloped viruses.

STUDY DESIGN AND METHODS: Suspensions of B19 in coagulation factor concentrates (FVIII) were irradiated with UVC light. B19 infectivity was determined by an indirect immunofluorescence assay using CFU–E, as a host cell, derived from peripheral blood CD34+ cells. The effects of catechins on B19 infectivity and on FVIII activity after UVC illumination were also examined.

RESULTS: The indirect immunofluorescence assay estimated the B19 infectivity of samples containing virus copies of 105 to 1011 per 10 μL to be a median tissue culture‐infectious dose of 100.3 to 105.4 per 10 μL. B19 was inactivated by 3 log at 750 J per m2 of UVC radiation and was undetectable after 1000 or 2000 J per m2 of irradiation. However, FVIII activity decreased to 55 to 60 percent of pretreatment activity after 2000 J per m2 of UVC radiation. This was inhibited in the presence of rutin or catechins. Epigallocatechin gallate could maintain FVIII activity at almost 100 percent of pretreatment activity after 2000 J per m2 of UVC radiation, while B19 infectivity was decreased to undetectable levels, which resulted in >3.9 log inactivation.

CONCLUSION: UVC radiation in the presence of catechins, especially epigallocatechin gallate, appears to be an effective method of increasing the viral safety of FVIII concentrates without the loss of coagulation activity.
Monitoring for underutilization of RBC components and platelets
Authors: Saxena Sunita , Wehrli Gail , Makarewicz Kathleen , Sartorelli Jennifer , Shulman Ira A.
Published: March 2003

BACKGROUND: Monitoring blood transfusion for overutilization is standard practice at most institutions.

STUDY DESIGN AND METHODS: This study monitored for underutilization of blood transfusion over a 14‐month period, by evaluating patients who had Hb levels that were reported to be <5 g per dL or platelet counts <10 × 109 per L and who did not receive an RBC or platelet transfusion within 24 hours of the reported results.

RESULTS: During the study period, 24,004 units of RBCs and 3,967 units of apheresis platelets were transfused. There were 148 patients who had a Hb level that was reported to be <5 g per dL or a platelet count reported to be <10 × 109 per L and who did not receive a transfusion during the 24 hours after the reporting of these results. In 5 cases, the patients died before the reporting of the low Hb or platelet counts, which precluded the low Hb or low platelet count reports from triggering transfusion therapy. In 8 cases, an underutilization review investigation could not be done, because of the unavailability of patient charts. Of the remaining 135 cases, investigation revealed justifiable reasons for withholding transfusion in 133. In 2 cases, the withholding of transfusion was deemed by peer review to be inappropriate, as the patients should have received a transfusion. Overall, there was one documented underutilization of RBC transfusion therapy during a period when 24,004 units were transfused and one underutilization of platelet transfusion therapy during a period when 3,967 units of apheresis platelets were transfused.

CONCLUSION: Monitoring for underutilization of transfusion therapy fulfills the requirements of the Joint Commission on the Accreditation of Healthcare Organizations. While the underutilization of transfusion therapy did not appear to be a significant problem at this medical center, determining the reasons for withholding transfusions shed light on important patient care‐related issues, including preexisting causes of falsely low platelet counts and Hb levels, delays in investigating critical laboratory values, and the need for policies for the treatment of patients who refuse transfusion for personal or religious reasons.
Effects of colloidal resuscitation fluids on the neutrophil respiratory burst
Authors: Jaeger Karsten , Heine Joern , Ruschulte Heiner , Jüttner Björn , Scheinichen Dirk , Kuse Ernst R. , Piepenbrock Siegfried
Published: March 2003

BACKGROUND: Experimental studies have revealed that gelatin and HES produce increased neutrophil respiratory burst activity. It was investigated whether 3‐percent gelatin (MW 35,000) and three types of 6‐percent HES (MW 70,000; degree of substitution, 0.5; 200,000/0.5; 450,000/0.7) preparations can influence superoxide anion production during respiratory burst under clinical conditions.

STUDY DESIGN AND METHODS: Blood samples were obtained from 40 patients before and 1 hour after the infusion, before anesthesia and surgical treatment. After stimulation with bacteria (Escherichia coli), the respiratory burst was measured by oxidation of nonfluorescent dihydrorhodamine 123 to the fluorescent rhodamine 123 by the use of flow cytometry.

RESULTS: Respiratory burst activity decreased significantly (p = 0.004) from the baseline (60.0 ± 6.5%) to 1 hour after the administration of the low‐molecular‐weight HES preparation (55.0 ± 6.8%). No significant differences in respiratory burst activity could be found after the administration of gelatin or medium‐molecular‐weight or high‐ molecular‐weight HES solution.

CONCLUSION: The investigated administration of gelatin and medium‐ and high‐molecular‐weight HES preparations did not influence respiratory burst activity under clinical conditions. However, the neutrophil respiratory burst was impaired after the administration of low‐molecular‐weight HES. Neutrophil respiratory burst activity may vary according to the type of colloidal plasma substitutes administered.
Duck hepatitis B photoinactivation bydimethylmethylene blue in RBC suspensions
Authors: Wagner Stephen J. , Skripchenko Andrey , Pugh John C. , Suchmann Donna B. , Ijaz M. Khalid
Published: March 2003

BACKGROUND: Dimethylmethylene blue (DMMB) has been used to photoinactivate a number of model viruses, including VSV, in RBC suspensions under conditions that preserve in vitro RBC properties during storage.23 The relative sensitivity of duck HBV (DHBV) and VSV to photoinactivation by DMMB was investigated by performing an indirect immunofluorescence assay (IFA) using primary duck hepatocyte (PDH) cultures or a standard plaque assay for the respective viruses.

STUDY DESIGN AND METHODS: DMMB was added to 45‐percent Hct, WBC‐reduced, oxygenated AS‐3 RBCs at 10‐, 1‐, and 0.1‐μM concentrations. Samples (1‐mm thick) were illuminated with 5.4‐mW per cm2 of red light for 2 or 9 seconds. Unilluminated samples without DMMB or with 10 μM DMMB served as control.

RESULTS: DHBV and VSV were rapidly photoinactivated by DMMB in a concentration and light‐dose‐dependent fashion. Neither virus was substantially inactivated by incubation with DMMB in the dark. For a given light exposure, DHBV required a concentration of DMMB one‐one hundredth that of VSV to achieve approximately the same level of inactivation.

CONCLUSION: DHBV appears to be considerably more sensitive than VSV to DMMB photoinactivation. Photoinactivation in 45‐percent Hct RBCs can be achieved in seconds by using micromolar quantities of dye.
UV‐visible spectrophotometric approach to blood typing II: phenotyping of subtype A2 and weak D and whole blood analysis
Authors: Narayanan Smita , Galloway Lamar , Nonoyama Akihisa , Leparc German F. , Garcia‐Rubio Luis H. , Potter Robert L.
Published: March 2003

BACKGROUND: A recently introduced quantitative blood typing approach uses antibody‐induced changes in the UV‐visible spectra of blood. Changes in the blood spectra's slope, caused by RBC agglutination, are translated into a numerical agglutination index (AI). Comparing the AI value against an established threshold yields a “yes and/or no” output from which to determine the phenotype. The efficacy and flexibility of this approach with whole blood use and the ability to analyze weak D, A2, and A2B were examined.

STUDY DESIGN AND METHODS: Two hundred randomly selected blood bank donor samples were coded and forward typed directly from whole blood by using the spectrophotometric analysis. Reverse grouping on plasma from each sample was carried out with a new modified procedure by using higher ratios of plasma to RBCs. Results were compared to typing by an FDA‐cleared automated typing system. Twenty‐seven weak D samples, 15 A2 and 12 A2B, were similarly analyzed from whole blood. PEG improved detection of weak D, A2 and A2B subtypes.

RESULTS: All two hundred coded samples were accurately typed, yielding identical results to the blood bank analysis in both forward and reverse grouping. All the weak D samples and A2 and A2B samples were clearly identified, having AIs above the type threshold indicator value.

CONCLUSION: Spectrophotometric blood typing successfully phenotyped ABO and D in 200 whole blood samples. Reverse grouping of plasma was equally successful. The same method can identify weak D and A2 and A2B subtypes.
A randomized controlled trial comparing the frequency of acute reactions to plasma‐removed platelets and prestorage WBC‐reduced platelets
Authors: Heddle Nancy M. , Blajchman Morris A. , Meyer Ralph M. , Lipton Jeff H. , Walker Irwin R. , Sher Graham D. , Constantini Lorrie A. , Patterson Bruce , Roberts Robin S. , Thorpe Kevin E. , Levine Mark N.
Published: March 2003

BACKGROUND: Removal of the plasma supernatant from platelets before transfusion is effective in preventing acute reactions to platelets caused by cytokines. Prestorage WBC reduction of platelets may be even more effective at preventing reactions as the WBCs are removed and WBC‐derived cytokines do not accumulate in this component. This study evaluates the effectiveness of plasma removal and two methods of prestorage WBC reduction for preventing acute reactions to platelets.

STUDY DESIGN AND METHODS: Platelets given to adults with hematologic malignancies were randomly allocated to one of three types: plasma supernatant removed and a platelet storage solution added, whole blood‐derived platelets that are prestorage WBC reduced by filtration before storage, and prestorage WBC‐reduced apheresis platelets. Patients were monitored before, during, and after transfusion, and the severity of reactions was graded on a Likert scale.

RESULTS: A total of 129 patients from four centers were given 1190 platelet transfusions. The overall frequency of reactions was 13.6 percent (162 of 1190), 21.3 percent (36 of 169) for the plasma‐removed platelets, 11.4 percent (59 of 517) for random donor WBC‐reduced platelets, and 13.3 percent (67 of 504) for apheresis WBC‐reduced platelets (p = 0.384). The overall frequency of severe reactions was 4.1 percent with plasma‐removed platelets, 1.7 percent for whole blood‐derived, prestorage WBC‐reduced platelets, and 1.4 percent for prestorage WBC‐reduced apheresis platelets.

CONCLUSION: The frequency of reactions to plasma‐removed platelets and prestorage WBC‐reduced platelets was not significantly different; however, the power of the study for this comparison was low. There was no difference in the frequency of reactions to the two types of prestorage WBC‐reduced platelets. The frequency of severe reactions to prestorage WBC‐reduced platelets is low, occurring in only 1 to 2 percent of transfusions.
Strategies for diagnosis of HTLV‐I and ‐II
Authors: Thorstensson Rigmor , Albert Jon , Andersson Sören
Published: March 2003

BACKGROUND : No gold standard exists for diagnosis of HTLV infection. The aim of thus study was to compare the accuracy of a combination of two sensitive ELISAs with Western blot (WB), a line immunoassay, and PCR for diagnosis of HTLV infection.

STUDY DESIGN AND METHODS : Nine hundred eighty‐five specimens were tested for the presence of HTLV antibodies by HTLV‐I and/or HTLV‐II EIAs (Murex and Ortho), WB (Diagnostic Biotechnology), line immunoassay (INNO‐LIA, Innogenetics), and/or presence of HTLV DNA by PCR. The results were compared with the probable HTLV infection status of each subject, as determined by detailed review of all available laboratory, clinical, and epidemiologic data.

RESULTS : The sensitivity for diagnosis of HTLV‐I infection was high for all assays evaluated, but both PCR and WB had a lower sensitivity rate (approx., 80%) for confirmation of HTLV‐II. INNO‐LIA detected 94 percent of the HTLV‐II‐positive samples. However, Murex EIA in combination with Ortho EIA was 100‐percent sensitive for the detection of both HTLV‐I and HTLV‐II antibodies. Furthermore, the number of samples giving indeterminate results in the ELISA combination was much lower as compared with WB (2.5% vs. 50%).

CONCLUSION : Based on these findings, a new, more sensitive and specific test strategy for HTLV diagnosis than the current algorithm, which includes WB, is proposed. Thereby, both the direct and indirect costs can be substantially reduced.
Inactivation of parvovirus B19 during pasteurization of human serum albumin
Authors: Blümel Johannes , Schmidt Ivo , Willkommen Hannelore , Löwer Johannes
Published: March 2003

BACKGROUND : It has been shown that HSA may be contaminated with parvovirus B19 (B19) DNA. However, the presence of B19 DNA does not necessarily indicate infectious virus. HSA is pasteurized at 60°C for 10 hours and it remains unclear whether this procedure inactivates B19. Studies with animal parvoviruses indicate considerable heat resistance at 60°C. However, due to the lack of a suitable cell culture system, the pasteurization process has not been investigated in the past.

STUDY DESIGN AND METHODS : The recently described cell clone KU812Ep6 was used to establish a system for investigation of B19 inactivation during pasteurization. Virus‐infected cells were detected by immunofluorescent staining of viral capsid antigen and by RT‐PCR assay of virus‐specific capsid mRNA.

RESULTS
: B19 was inactivated after 10 minutes at 60°C for ≥4 log
. In contrast, porcine parvovirus was widely resistant at 60°C. Inactivation of B19 was independent of the analyzed albumin products (5, 20, and 25% albumin from three manufacturers) and from the specific virus source used for the inactivation experiments. Degradation of B19 DNA by deoxyribonuclease I treatment after pasteurization indicated that the virus capsid is destroyed during heat treatment.

CONCLUSION : Heat resistance of B19 markedly differs from heat resistance of animal parvoviruses. While animal parvoviruses widely withstand pasteurization of albumin, B19 was rapidly inactivated. These results confirm the safety of pasteurized albumin and are in line with its good clinical safety record with respect to B19 infection. However, conclusions regarding the safety of other blood‐derived medicinal products should not be derived from B19 inactivation in albumin, because different processes or different composition of product intermediates may significantly influence B19 stability during heat treatment.