bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical Evaluation of Transfusion of Prestorage‐Leukoreduced Apheresis Platelets
Authors: Ogawa Y. , Wakana M. , Tanaka K. , Oka K. , Aso H. , Hayashi M. , Seno T. , Ishida T. , Nomura S. , Fukuhara S.
Published: December 2004

With increased use of platelet concentrate (PC) in recent years, adverse reactions to PC transfusion have received much clinical attention. Most of these reactions stem from white blood cells (WBC) contaminating the transfused PC. Several are thought to be preventable by removing WBC before PC storage. Methods: We routinely filtered apheresis PC either during collection or immediately afterwards and monitored various indicators of platelet quality during storage. After transfusion to patients, transfusion efficacy and the type, severity, and frequency of posttransfusion side effects were compared with those of a control group in which PC was filtered at the bedside. Results: Prestorage‐filtered PC contained an average of 3.1±0.7×1011 platelets and 0.9±1.2×106 contaminating leukocytes. Messurement of platelet function and metabolic indicators revealed normal values throughout the storage period. CD62 measurement revealed no undue platelet activation after filtration or during the storage period. Cytokine, histamine, bradykinin, and complement levels showed no significant increase after filtration or during storage. Transfusion efficacy and overall side effect incidence rates were similar for the prestorage‐and bedside‐filtered groups, but reactions of the bedside‐filtered group included serious reactions such as breathing difficulties and shock. No serious reactions were noted in the prestorage‐filtered group. Conclusion: Filtering PC before storage has no adverse effect on PC quality and may reduce the severity of post transfusion side effects.
Prevention of Cytokine Accumulation in Platelets Obtained with the COBE Spectra Apheresis System
Authors: Palmer D. S. , Aye M. T. , Dumont L. , Dumont D. , McCombie N. , Giulivi A. , Rutherford B. , Trudel E. , Hashemi‐Tavoularis S.
Published: December 2004

Background and Objectives: Febrile nonhemolytic transfusion reactions frequently accompany platelet trasfusions and may be due to accumulation of cytokines mediating inflammation during storage of platelet concentrates (PCs). We wished to determine whether PCs collected using the COBE® spectra™ Apheresis System (Version4) were sufficiently leukocyte reduced (LR) to limit cytokine accumulation during storage. Materials and Methods: Cytokine accumulation‐interleukin (IL)‐1β, IL‐6, IL‐8, tumor necrosis factor‐α (TNF‐α)‐ and release of platelet α‐granule ‐ P‐selectin, transforming growth factor‐β1 (TGF‐β1), platelet‐derived growth factor AB (PDGF‐AB), von Willebrand factor (vWf)‐ or dense granule (serotonin) markers were investigated during a 7‐day storage period comparing apheresis‐collected, LR PCs (LR PCs) and random donor platelets prepared from whole blood (WB). Results: Leukocyte counts were reduced 99.95% comparing LR PCs (5.7×105/1) and WB PCs (1.09×109/1). Little or no accumulation of leukocyte‐derived cytoknies was observed in LR PCs during storage in contrast to WB PCs. A reduction in the release of platelet α‐granule proteins, such as P‐selectin, TGF‐β1 and PDGF‐AB, was observed on day 0 for LR PCs compared to WB PCs with little or no difference observed from day 3 to 7. Plasma vWf levels were higher in LR PCs compared to WB PCs on days 0–7. Conclusion: Leukocyte levels in PCs collected with the COBE Spectra Apheresis System are sufficiently low to limit cytokine production during 7 days of storage.
c‐fos and c‐jun mRNA Expression in Activated Cord and Adult Lymphocytes: An Analysis by Northern Hybridization
Authors: DePalma L. , Brown E. , Baker R.
Published: December 2004

Background and Objectives: To further analyze the neonatal immune response to an antigenic challenge such as blood transfusion, c‐fos and c‐jun mRNA expression were analyzed in twelve in‐vitro‐stimulated normal cord blood and ten in‐vitro‐stimulated normal adult peripheral blood lymphocyte samples. Materials and Methods: Lymphocyte samples were stimulated by either the mitogen phytohemagglutinin (PHA) or the monoclonal antibody αCD3. Proliferation rate and Northern blot hybridization were employed. Results: Cord lymphocytes revealed a greater proliferation rate with PHA and αCD3 than adult lymphocytes (p=0.0081 and 0.0023, respectively). In addition, Northern blot analysis of cord and adult samples revealed similar maximal increases in c‐fos (99±15 and 126±11%, p=0.0126) and c‐jun (123±9 and 185±38%, p=0.0291) mRNA expression, respectively, as early as 15 min postαCD3 stimulation. Adult lymphocytes showed an equivalent increase in mRNA expression of c‐fos and c‐jun (140±25 and 155±31%) at 30 min post‐PHA stimulation, while cord lymphocyte maximum c‐fos and c‐jun expression (82±6 and 142±12%) occurred at 15 min post‐PHA stimulation (c‐fos, p=0.0354; c‐jun, p=0.0112). Conclusion: Although cord lymphocyte proliferation rates were significantly greater than those of adult lymphocytes following stimulation, lymphocyte activation, as analyzed by c‐fos and c‐jun mRNA expression, appears similar in both cord and adult samples. We conclude that cord lymphocyte activation exhibits an adult‐type profile.
Intrapatient Comparison of an Intermittent and a Continous Flow Cell Separator for the Collection of Progenitor and Stem Cells from the Blood
Authors: Beretta Fabrizio , van den Bosch Sabine , Castelli Damiano , Cavalli Franco , Ghielmini Michele
Published: December 2004

Background and Objectives: Continuous‐flow and intermittent‐flow blood cell separators (CFCS and IFCS) are both used to collect stem cells from the blood to rescue patients undergoing myeloablative treatment for cancer. Materials and Methods: We designed a study to compare the collection efficiency of the two systems. The continous‐flow Cobe Spectra and the intermittent‐flow Haemonetics MCS‐3P were used to collect cells on consecutive days from 9 patients mobilised with G‐CSF with or without chemotherapy. Blood obtained before leukapheresis and the leukapheresis product were analysed for their content of red and white cells, platelets, CD34‐positive cells, GM‐CFC, CFC‐E, and BFU‐E. An extraction ratio was calculated. Results: We found that the CFCS extracted about 4 times more mononuclear cells per unit time, 3 times more CD34‐positive, and 4 times more clonogenic cells than the IFCS. The subject acceptability of the two systems was similar. Conclusion: The CFCS is a more efficient system for stem cell collection. IFCS requires a longer harvesting time for the same result.
Creutzfeldt‐Jakob Disease and the Risk from Blood or Blood Products
Authors: Will R. G. , Kimberlin R. H.
Published: December 2004

The occurrence of iatrogenic cases of Creutzfeldt‐Jakob disease (CJD) and the isolation of infectivity in some laboratory transmission studies in transmissible spongiform encephalopathies raises the possibility that CJD might be accidentally transmissible through blood or blood products. Epidemiological evidence, although not conclusive, does not suggest that classical CJD is transmitted through this route. However, new variant CJD (nvCJD) might pose greater risks of accidental transmission of infection and mechanisms to reduce the theoretical risk are under consideration. The theoretical risks from CJD and nvCJD must be balanced against the establised therapeutic benefits of blood and blood products.
Erythrocytes and Proinflammatory Mediators in Wound Drainage
Authors: Mottl‐Link S. , Russlies M. , Klinger M. , Seyfarth M. , Ascherl R. , Gradinger R.
Published: December 2004

Background and Objectives: Retransfusion of shed blood collected after operation has become popular, but recent reports of side effects led to a search for possible causes. Materials and Methods: In a randomized study of 28 patients undergoing total hip arthroplasty, shed blood was collected in Solcotrans, Orth Evac, and ordinary Redon drainage. Osmotic fragility was measured and electron‐microscopic pictures of erythrocytes from selective samples were taken. Serotonin, prostaglandin E2 (PGE2), and histamine were measured with enzymelinked immunosorbent assays. Results: Higher osmotic fragility of erythrocytes collected with Solcotrans appeared to be due to ACD which was used only with that system. Serotonin concentrations did not differ significantly. However, there was a great increase in histamine (Solcotrans 477.7, Orth‐Evac 344.0, Redon drainage 453.1 nmol/ml) and PGE2 (Solcotrans 1,908.3, Orth‐Evac 1,225.0, Redon drainage 2,666.7 μg/ml) in shed blood compared with venous blood (histamine 9.5 nmol/l, PGE2 4.2 μg/ml). Conclusion:Unwashed wound drainage blood collected after operation contains levels of proinflammatory mediators that can account for the reported side effects.
Evaluation of in vivo and in vitro Quality of Apheresis‐Collected RBC Stored for 42 Days
Authors: Holme S. , Dean Elfath M. , Whitley P.
Published: December 2004

Background and Objectives: New technological developments make it possible to collect red blood cells (RBCs) by apheresis, which allows for better product consistency and has the potential for improved RBC quality. The purpose of these studies was to evaluate the quality and consistency of units of RBCs collected by apheresis using the MCS+® machine (Haemonetics Corp., Braintree, Mass., USA). Materials and Methods: Two studies were performed. In study 1 (n = 10), using containers and CP2D/AS‐3 solutions from Medsep Corp. (Covina, Calif. USA), one‐unit apheresis RBCs were compared to manually collected RBCs in a random crossover design. In study 2 (n = 12), 6 subjects had one unit collected, while the remaining 6 subjects had two units of RBCs collected with comparison to previously manually collected RBCs from the same donors. Haemonetics containers and solutions were used in study 2. Results: Low RBC volume variability was found for the apheresis collections with a standard deviation of only 6 ml difference between actual and target volumes. Combining the data from the two studies (n = 21 pairs), at 42 days of storage, the apheresis units showed slightly lower hemolysis (0.44±0.26 vs. 0.61±0.50%), lower supernatant potassium levels (50±3 vs. 53±3 mEq/l), and improved tolerance to osmotic shock (47±3 vs. 49±3%) as compared to manual units (p < 0.05). There was no statistically significant difference in RBC ATP (3.0±0.6 vs. 2.9±0.5 μmol/g Hb) or in 24‐hour percent recoveries (81±6 for apheresis vs. 81±4% for apheresis red cells). Apheresis RBC quality was not affected by the manufacturer (Haemonetics vs. Medsep) of solutions and containers. Conclusions: RBC units collected by apheresis demonstrated low variability in volume of RBC mass collected, and showed similar RBC properties as compared to manually collected RBCs after processing and after 42 days of storage.
A Stable Reagent System for Screening and Identifying Red Blood Cell Irregular Antibodies: Application to Commercial Antibodies
Authors: Million L. , Pellerin C. , Marchand‐Arvier M. , Vigneron C.
Published: December 2004

Objective: Development of a new solid‐phase system for screening and identifying irregular red cell antibodies. Materials and Methods: Red blood cell membranes were prepared by a semi‐automated procedure in which the hemolysate solution was passed through a hollow‐fiber system. The membranes were fixed to the solid phase (microtiter plates) by centrifugation and incubated with 8% fat‐free milk. Antibodies added to the microtiter plate were detected by antihuman antibodies adsorbed onto yellow latex particles. Results: The system had good sensitivity (titer <1); 97% of anti‐D samples were detected. The detection system was stable for 6 months at 4°C. Conclusion: This stable‐antigen solid‐phase system readily detects and identifies red cell antibodies that are important in transfusion.
Detection of HIV‐1 RNA in Two Consecutive Blood Donations Screened Negative for HIV Antibodies
Authors: Burin des Roziers Nicolas , Coste Joliette , Courouce Anne‐Marie , Bibollet‐Ruche Frédéric , Guillard Alain , Nasr Olivier
Published: December 2004

Background and Objectives: An unusual serological pattern of HIV‐1 seroconversion in a blood donor is described. The seroconversion panel was used to investigate the sensitivity of existing screening assays. Materials and Methods: A volunteer blood donor who had given blood 79 times was diagnosed anti‐HIV‐1‐antibody‐positive. The heteroduplex mobility assay identified a subtype B HIV‐1 strain. The frozen plasmas from the last four blood donations had been kept at −30°C. They were thawed and aliquoted for subsequent testing. Results: The last two blood donations contained HIV‐1 RNA, 2,800 copies/ml (October 26) and 170 copies/ml (November 23). Weak anti‐p24 antibodies were detected by Western blot in the October 26 sample, and a clear p24 reactivity along with a faint gp 160 reactivity was oberved on November 23. HIV p24 antigen was undetectable in both samples. Out of 13 screening assays, only 6 gave positive results on the November sample and 7 negative results which were obtained by 1 competitive enzyme immunoassay (EIA) and 6 of the 9 sandwich EIAs. Conclusion: Most sandwich EIAs gave prolonged false‐negative results in the present case. p24 antigen testing was negative and would not have reduced the risk of HIV transmission.
Efficiency of Leukocyte Removal by Filters Made of Superfine Glass Fiber Membranes
Authors: Yuanguo Zou , Qijun Sun , Aihua Li , Fangfang Yao , Zhengfang Hu , Zhen Li , Wenrong Ma
Published: December 2004

Background and Objectives:
To demonstrate the application of leukocyte removal filters made of a new type of filter material – superfine glass fiber – for depleting leukocytes in SAGM red cell suspensions and preventing nonhemolytic transfusion reactions. Materials and Methods: The extent of leukocyte depletion and red cell recovery was based on cell counts. Trace leukocytes were counted in a 50‐μl Nageotte counting chamber or by using a flow cytometer. The chemical stability of the glass fiber membranes was studies by plasma emission spectrometer and by measuring the ion content and weighing nonvolatile matter in water extract. The structural stability of the glass fiber membranes was studied by a micropore‐filter membrane method. Results: Leukocyte removal filters made of superfine glass fiber membranes removed more than 99.0% of leukocytes in SAGM red cell suspensions prepared from 400 ml whole blood. Red cell recovery exceeded 90%, and the total number of residual leukocytes was less than 5 × 106. A water extract of the glass fiber membranes contained only traces of Si4+ and Ca2+ and less than 2 mg/100 ml of nonvolatile matter. No broken or loose fibers were found in the filters. Scanning electron microscopy showed that the web structure of the glass fiber membranes was instrumental in trapping and holding leukocytes. Conclusion: A filter made of glass fiber membranes is effective in leukocyte depletion.
A Very‐High‐Purity von Willebrand Factor Preparation Containing High‐Molecular‐Weight Multimers
Authors: Barington Karina A. , Kærsgaard Per
Published: December 2004

Background and Objectives:
We investigated largescale production of very‐high‐purity von Willebrand factor (vWf) containg high‐molecular‐weight vWf multimers, using chromatography alone. Materials and Methods: Factor VIII (FVIII)‐containing vWf was produced by intial separation of vWf from plasma by gel filtration followed by two ion exchange steps. two virus inactivation steps were incorporated in the process. Results: The process resulted in FVIII‐containing vWf preparations with a mean specific activity of 82 U vWf:collagen‐binding activity per milligram protein (excluding added albumin) and with almost intact vWf multimer distributions. Conclusion: Very‐high purity, double virus‐inactivated vWf preparation containing high molecular weight multimers were obtained on a production scale.
Search for the Sialic Acid‐Independent Receptor on Red Blood Cells for Invasion by Plasmodium falciparum
Authors: Soubes Sandrine C. , Reid Marion E. , Kaneko Osamu , Miller Louis H.
Published: December 2004

Background and Objectives:

Plasmodium falciparum uses multiple red blood cell (RBC) receptors and parasite ligands to invade RBCs. One pathway uses a sialic acid‐independent protein or carbohydrate for invasion. The present study searches for this RBC receptor. Materials and Methods: We determined whether antigennegative and null RBCs (including PNH cells that lack all glycosylphosphatidyl inositol‐linked proteins) could be invaded after neuraminidase treatment. We used two P. falcipa clones for the study: one that requires sialic acid for invasion and was an indication of removal of sialic acid and a second clone that can invade neuraminidase‐treated RBCs. Results: All neuraminidase‐treated variant RBCs in this study were invaded. Conclusion: This study indicates that some molecule other than those studied (e.g., a carbohydrate) is the receptor for the sialic acid‐independent pathway. This powerful tool for the identification of receptors for microorganisms should be used more extensively.
Prevalence of GBV C/HGV RNA and GBV C/HGV Antibodies in French Volunteer Blood Donors: Results of a Collaborative Study
Authors: Mercier B. , Barclais A. , Botte C. , Cantalube J.F. , Coste J. , Defer C. , Gautreau C. , Giannoli C. , Halfon P. , Lepot I. , Loiseau P. , Martial J. , Montcharmont P. , Merel P. , Ouzan D. , Ravera N. , Follana J. , Césaire R. , Janot C. , Lemaire J.M. , De Micco P. , Vezon G. , Férec C.
Published: December 2004

Background and Objectives:
Posttransfusion hepatitis still occurs at an incidence of about 1 in 118,000 for HBV and 1 in 220,000 for HCV. This collaborative study aimed to determine the prevalence of a novel flavivirus, GBVC/HGV, even though its role in transfusion‐associated hepatitis is uncertain. Materials and Methods: GBVC/HGV RNA was detected by PCR using either the Boehringer detection kit or by primers previously described. HGV antibodies were detected by a serological assay from Boehringer. Results: The observed GBVC/HGV RNA frequency was 3.4%. HGV antibodies occurred in 9.5% of donors. Conclusion: In our study, 12.9% of the donors had been in contact with the GBVC/HGV virus.
Decreased Hemolysis and Lipid Peroxidation in Blood during Storage in the Presence of Nicotinic Acid
Authors: Arun P. , Padmakumaran Nair K.G. , Manojkumar V. , Deepadevi K.V. , Lakshmi L.R. , Kurup P.A.
Published: December 2004

Background and Objectives:
There is increase in lipid peroxidation with consequent increase in hemolysis when blood is stored in di‐(2‐ethyl hexyl)phthalate (DEHP) plasticized bags. Studies carried out by us and others have indicated the ability of red cells to synthesize NAD+ from added nicotinic acid. Apart from the role of NAD+ in glycolysis, NADPH is required for reduction of oxidized glutathione to its reduced form by glutathione reductase. Reduced glutathione is an important antioxidant, which protects cell membrane from oxidative damage. Reduced glutathione is also involved in the regeneration of vitamin E, another important membrane antioxidant. In view of these, a study was undertaken to find out the effect of addition of nicotinic acid to the citrate‐phosphate‐dextrose‐adenine (CPDA) solution on lipid peroxidation and integrity of red cells when whole blood is stored in DEHP plasticized bags. Materials and Methods: Blood was collected in Penpol blood storage bags (which is a DEHP plasticized bag) in CPDA solution in the presence and absence of nicotinic acid. Various parameters of lipid peroxidation and membrane stability – level of malondialdehyde (MDA), conjugated dienes, vitamin E, reduced glutathione, plasma Hb and K+, levels of adenosine triphosphate (ATP) and 2,3 diphosphoglycerate (2,3‐DPG) were studied in the blood samples after various periods. Results: Plasma Hb and K+ concentrations were significantly lower in the presence of added nicotinic acid both after 28 and 42 days. Concentration of MDA and conjugated dienes was lower and the levels of reduced glutathione and vitamin E higher in the presence of nicotinic acid. ATP levels were not significantly different, but 2,3‐DPG levels were higher. pH of the blood was nearer to 7.0 in the presence of nicotinic acid, while leaching out of DEHP into the blood was significantly lower. Conclusion: Inclusion of nicotinic acid in the CPDA solution has a beneficial effect in that (1) it reduces plasma Hb and K+; (2) reduces lipid peroxidation and increases antioxidant protection; (3) maintains pH nearer to 7.0, and (4) decreases the leaching out of DEHP into the blood.
Preparation of Leukodepleted Platelet Concentrates from Pooled Buffy Coats: Prestorage Filtration with Autostop™BC
Authors: Pietersz R.N.I. , van der Meer P.F. , Steneker I. , Hinloopen B. , Dekker W.J.A. , van Zanten A.P. , Reesink H.W.
Published: December 2004

Background and Objectives:
Our requirements for leukocyte‐depleted platelet concentrates (LD‐PC) for an adult patient are: platelets >240×109, leukocytes <5×106, volume of 150–400 ml; and at the end of storage a pH between 6.8 and 7.4 and presence of the swirling effect. Our aim was to develop a standardized, semiautomated method for the production of LD‐PC, by pooling of buffy coats (BC), and prestorage leukoreduction by filtration. Materials and Methods: Whole blood was collected in Top and Bottom systems, and separated automatically with the Compomat™ G3 equipment into a red cell concentrate, a plasma and a BC. Subsequently, a pool of 5 BC was made, and 200 g plasma from one of the donors was added. Then, after soft spin centrifugation, the platelet rich plasma was leukocyte depleted by filtration using the Autostop™ BC filter, and stored in a 1,000 ml polyolefin platelet storage bag. Results: BC (n = 60) had a volume of 51±2 ml (mean ± SD) with a hematocrit of 0.44±0.03 I/I and contained 80±5% of the platelets and 74±12% of the leukocytes of the whole blood. Routinely prepared LD‐PC (n = 15,037) contained a median of 341×109 platelets (range 49‐599×109), with only 104/15,037 (0.7%) containing fewer than 240×109 platelets; the median volume was 263 ml (range 134‐373 ml). In 118/917 (13%) LD‐PC leukocytes were observed in the Nageotte hemocytometer, but only twice exceeding 1×106 leukocytes per unit, and none exceeding 5×106 (median <0.6×106; range <0.6‐1.41×106). Storage experiments of the LD‐PC (n = 12) revealed adequate oxygenation and maintenance of pH and swirling effect up to 9 days. Conclusions: This method warrants with 99% confidence that LD‐PC contain more than 240×109 platelets; with 97.5% confidence that 100% of the LD‐PC contain <5×106 leukocytes, and with 95% confidence that more than 99% of the LD‐PC contain fewer than 1×106 leukocytes; these LD‐PC can be stored satisfactorily for up to 9 days.
Processing of Human Cord Blood by Three Different Procedures for Red Blood Cell Depletion and Mononuclear Cell Recovery
Authors: Perutelli Paolo , Catellani Silvia , Scarso Lucia , Cornaglia‐Ferraris Paolo , Dini Giorgio
Published: December 2004

Background and Objectives:
Human cord blood (CB) is an important source of stem cells which may be used for hematopoietic reconstitution as an alternative to bone marrow transplantation. Banking of CB would be accomplished by removing red blood cells (RBC) and plasma from CB collections. Our aim was to compare three different procedures for CB processing. Materials and Methods: Poligeline, hydroxyethyl starch gel (HES) and gelatin were used as separation media in processing 79 CB units for RBC depletion and mononuclear cell (MNC) recovery. Results: The best MNC recoveries were obtained performing the HES‐ and the gelatin‐based procedures (80.9 and 84.7%, respectively), but the gelatin procedure allowed us to obtain the highest RBC depletion (96.4%); CD34+ cell recovery was higher using HES or gelatin as separation media (85.6 and 85.9%, respectively). Conclusion: The best results, as far as RBC removal and MNC recovery are concerned, were obtained by using gelatin as RBC sedimentation medium. Gelatin is a low‐cost, animal‐derived reagent, which has been successfully used for CB transplantation; the procedure is simple to perform and appears to be suitable for large‐scale banking in view of CB transplantation.
Isoantibodies in Immunoglobulin for Intravenous Use May Cause Erythrocyte Sequestration
Authors: Shoham‐Kessary H. , Gershon H.
Published: December 2004

Background and Objectives:
IVIg, while generally a safe therapy for a variety of immunological disorders, can have detrimental effects on erythrocyte (RBC) homeostasis. We studied the mediation of erythrophagocytosis by isoantibodies in IVIg. Materials and Methods: RBC were exposed to IVIg and binding of IgG determined by flow cytometry. In vitro phagocytosis of these RBC was assayed. Results: Anti‐A and anti‐B in IVIg mediate Fc‐dependent erythrophagocytosis even in the presence of excess IVIg. Removal of these isoantibodies from IVIg prevents IgG binding and erythrophagocytosis. Complement enhances IVIg‐mediated RBC sequestration. Conclusion: It may be desirable to remove isoantibodies from IVIg especially for anemic patients or those who respond to the IVIg with a hemolytic episode.
Anti‐Granulocyte Antibody Screening with Extracted Granulocyte Antigens by a Micro‐Mixed Passive Hemagglutination Method
Authors: Araki Nobuo , Nose Yoshisuke , Kohsaki Masatoshi , Mito Hisashi , Ito Kazuhiko
Published: December 2004

Background and Objectives:
Serologic tests for granulocyte antibodies, i.e., the granulocyte agglutination test and the granulocyte immunofluorescence test, require panels of typed granulocytes that cannot be preserved for more than a few hours. We have developed a new method in which granulocyte antigens, extrcated into saline containing 3% sucrose, are coated onto U‐type Terasaki plates. With this new method, we evaluated the micro‐mixed passive hemagglutination test (EG‐MPHA) for screening for granulocyte antibodies. Materials and Methods: We tested the ability of the EG‐MPHA to detect granulocyte antigens using 5 human antibodies specific for NA1, NA2, NB1, 5b, and Sara, and 8 different monoclonal antibodies for NA1, CD11a, CD11b, CD13, CD16, CD18 and HLA class I. Sera from 94 alloimmunized patients were screened by the chloroquine‐treated EG‐MPHA method. Results: Na1, NA2, NB1, 5b, Sara, CD11a, CD11b, CD13, CD16, CD18 and HLA class I antigens were present in the extracted granulocyte antigen preparation. CD11b and HLA class I antigens were removed when the extracted granulocyte antigens were treated with chloroquine. Granulocyte antibody screening of sera from alloimmunized patients showed that approximately 30% of the anti‐HLA‐positive for granulocyte antibody by the chloroquine‐treated EG‐MPHA. The extracted granulocyte antigen panels could be stored frozen for at least 1 year at −80°C. Conclusion: This new method is preferable for screening for granulocyte antibodies. In addition, it has the advantage of requiring only 5 μl of serum for each test.
Press Review
Authors: Sunagawa T. , Uezo Y. , Kadena K.
Published: December 2004

Successful Treatment of a Non‐Haemophilic Patient with Inhibitor to Factor VIII by Double‐Filtration Plasmapheresis
Press Review
Authors: Goodnough L.T. , Brecher M.E. , Kanter M.H.
Published: December 2004

Transfusion Medicine, part 1

Transfusion Medicine, part 2
Oversight and Monitoring of Blood Safety in the United States
Authors: Busch Michael , Chamberland Mary , Epstein Jay , Kleinman Steven , Khabbaz Rima , Nemo George
Published: December 2004

The US blood safety vigilance system is composed of a network of interwoven programs, now organized under a formal structure, with the Assistant Secretary of Health and DHHS Blood Safety Committee bearing overall responsibility. It takes advantage of the breadth of expertise and close collaborative relationship of transfusion medicine and infectious disease scientists within and outside of the government. Core elements include an array of ongoing surveillance programs for monitoring established as well as new and emerging infectious agents that may pose a risk to blood safety, and the existence of historical and contemporary repositories of donor and recipient specimens that enable rapid investigation of putative new risks. This report summarizes the historical events that shaped the US blood safety oversight system, reviews the current organization and decision‐making processes related to blood safety issues, and highlights key surveillance systems and research programs which monitor the US and global blood supplies for known and potential emerging risks.
The French Haemovigilance System
Authors: Debeir Joëlle , Noel Luc , Aullen Jean‐Patrice , Frette Catherine , Sari Fatih , Vo Mai Mai Phuong , Cosson Alain
Published: December 2004

Haemovigilance is a national system of surveillance and alarm, from blood collection to the follow‐up of the recipients, gathering and analysing all untoward effects of blood transfusion in order to correct their cause and prevent recurrence. In France haemovigilance was created by law and notification of transfusion incidents is a legal obligation. The haemovigilance network associates local correspondents in each hospital and blood centre with regional co‐ordinators and is centralised by the Agence Francaise du Sang. After 4 years the incident reporting rate is 2.3 per 1000 allogeneic blood components transfused, justifying for example increased efforts in the prevention of bacteria‐associated transfusion reactions, haemolytic transfusion reactions or vascular overload. However, haemovigilance still has to be strenghtened by improved information management or further progress in standardisation from one region to the other. The most important factor of success is collaboration between blood centres and hospitals. Haemovigilance clearly is the ultimate quality indicator of a transfusion service.
Differences in Residual White Blood Cell Subset Counts in Buffy Coat‐Depleted Red Cell Concentrates Prepared with Bottom and Top or Quadruple‐Bag Systems
Authors: de Wildt‐Eggen J. , Schrijver J.G. , Kuiper‐Kramer P.A. , Bins M. , van Prooijen H.C.
Published: December 2004

Background: For preparation of buffy coat‐depleted red cell concentrates (RCCs) in additive solution whole blood is currently collected in The Netherlands both in quadruple‐bag and bottom and top bag systems. By using the quadruple‐bag system both plasma and buffy coat cells are transferred into integrated satellite bags while the red cells remain in the collection bag. When bottom and top bags are used, the buffy coat remains in the collection bag while both red cells and plasma are transferred into satellite bags. The difference in processing prompted us to perform quantitative analysis of residual WBC subsets in buffy coat‐depleted RCCs. Differences in removal of specific cells with the buffy coat could improve the outcome of additional filtration procedures aiming at complete removal of specific WBC subsets. Study Design and Methods: The buffy coat was removed in semiautomated procedures (Optipress I; Compomat G4) from units of whole blood collected in both bag systems. Paired samples were taken before and after removal of the buffy coat for counting and analyzing WBC subsets by flow cytometry using subset specific monoclonal antibodies. Results: All RCCs met the criteria from the guidelines of the Council of Europe. The percentage of residual total WBCs was lower (p < 0.001) in RCCs processed in bottom and top bag systems (26% Compomat and 18% Optipress) as compared to RCCs processed in quadruple‐bag systems (43% compomat). WBC subset analysis in RCCs processed in quadruple‐bag systems showed approximately 25% of residual T cells, B cells and monocytes and 60% of residual granulocytes. In contrast, WBC subset analysis in RCCs processed in bottom and top bag systems showed approximately 2% residual T cells, B cells, and monocytes and 35% residual granulocytes; in about 45% of units, lymphocytes and monocytes were even below the detection limit of flow cytometry analysis. Conclusion:Buffy coat‐depleted RCCs are currently processed in bottom and top bag or quadruple‐bag systems, the former being superior to the latter due to selective depletion of lymphocytes and monocytes by 98% (2 logs). The bottom and top procedure is an evident contribution to leukodepletion in blood transfusion, both with and without additional filtration.
An International Trial Demonstrates Suitability of a Newly Developed Whole‐Blood ELISA Kit for Multicentre Platelet HPA‐1 Phenotyping
Authors: Bessos Hagop , Hofner Maureen , Salamat Ahmed , Wilson David , Urbaniak Stan , Turner Marc L.
Published: December 2004

Background:
Neonatal alloimmune thrombocytopenia (NAIT) is in most cases due to pregnant women with HPA‐1b platelet phenotype producing antibodies to HPA‐1a platelets of the fetus, which may lead to intracranial haemorrhage with subsequent death or lifelong morbidity. The availability of sensitive, reliable, straightforward, and inexpensive assays would enable large‐scale screening in pregnancy and may help avoid NAIT. Methods: A recently developed enzyme‐linked immunosorbent assay (ELISA) was produced in kit form incorporating modified reagents and enabling distribution to 21 international Platelet Immunology and Blood Centres (see Acknowledgments). The kits were assessed using anticoagulated whole‐blood samples stored up to 10 weeks at 4°C. Each centre tested its own blood samples most of which had been previously typed by established assays. Results: Of the 152 samples that were tested, all 31 HPA‐1b phenotypes were correctly identified by the kit. The respective mean ± standard deviation of the specific absorbances of HPA‐1b and HPA‐1a samples were: 0.52±0.15 and 0.12±0.06 (p<0.0005). In addition, the modified ELISA showed 100% concordance with PCR‐SSP in the phenotyping of 93 donors. Finally, a comparison between freshly prepared and stored kits showed that the kit was stable for at least 2 years at 4°C. Conclusions: The international trial showed that the modified whole‐blood ELISA kit is very well suited for wide‐scale screening in pregnancy. Moreover, the ELISA kit could be used for large‐scale phenotyping of blood donors, with HPA‐1b‐typed individuals getting invited to become apheresis platelet donors for patients with NAIT.
Microsatellite Variation within the Human RHCE Gene
Authors: Kemp Tim J. , Poulter Mark , Carritt Ben
Published: December 2004

Background and Objectives:
This study provides a unique method for identifying individuals carrying the Rh haplotype cDe, and supports a model for the evolution of Rh haplotypes in which cDe is the progenitor. Materials and Methods: DNA from 212 unrelated donors of known Rh serological phenotype was PCR amplified. The resulting products were analysed by denaturing polyacrylamide gel electrophoresis, denaturing gradient gel electrophoresis and DNA sequencing. Results: Two adjacent microsatellite repeat elements of the form (AC)n (GCAC)n were found within the human Rh blood group genes. These display copy number variation which was non‐randomly distributed with respect to Rh serological phenotype, and was restricted to alleles of RHCE expressing the c antigen. Conclusion: The predominantly Black African allele cDe displayed a unique set of microsatellite alleles, providing a method of identifying individuals carrying this haplotype.
Press Review
Authors: Engelfriet C.P. , Munizzo M. , Nance S. , Keashen‐Schnell M.A.
Published: December 2004

Provision of HPA‐1a‐Negative Platelets for Neonatal Alloimmune Thrombocytopenia: Screening, Testing and Transfusion Protocol
Evaluation of the Safety of Rh Immunoglobulin by Monitoring Viral Markers among Rh‐Negative Female Blood Donors
Authors: Watanabe Kevin K. , Busch Michael P. , Schreiber George B. , Zuck Thomas F.
Published: December 2004

Background and Objectives:
Although immunoglobulin (lg) preparations including Rhlg have been noted for their record of safety, recent reports of hepatitis C virus (HCV) transmission by some lg preparations have raised concern. This analysis examined the safety of Rhlg manufactured in the US by comparing the prevalence and incidence of viral markers in Rh‐negative and Rh‐positive female blood donors. Materials and Methods: Demographic and viral marker data were analyzed for allogeneic donations collected from female donors of childbearing age (17–49 years of age) from April 1992 to May 1996. Prevalence and incidence rates were calculated for HCV, human immunodeficiency virus (HIV), and hepatitis B surface antigen (HBsAg). Results: Of the 624,939 female donors included in the study, 96,355 (15.4%) were Rh‐negative and 528,584 (84.6%) Rh‐positive. There were no significant differences in the prevalence of HCV and HIV between Rh‐negative and Rh‐positive female donors. HBsAg prevalence was significantly higher among non‐white compared to white donors. Following implementation of the more sensitive EIA 2.0 screening test for HCV in April 1992, prevalence and incidence rates declined over time at similar rates for Rh‐negative and Rh‐positive female donors. Conclusions: Rh‐negative female donors had similar prevalence and incidence rates for most viral markers compared to Rh‐positive female donors. This analysis supports the historical safety of Rhlg.
Leukocyte Depletion of Red Cell Components Prevents Exposure of Transfusion Recipients to Neutrophil Elastase
Authors: Willy C. , Reithmeier W. , Kuhlmann W.D. , Gerngross H. , Flegel W.A.
Published: December 2004

Background:
Polymorphonuclear leukocytes contain a large number of enzymes and bactericidal proteins stored in granules. Neutrophil activation induces degranulation and immediate release of these bioactive substances, including human neutrophil elastase (HNE) also know as elastase‐2 (ELA2), which may contaminate whole blood units and blood components. Materials and Methods: The HNE concentration was determined in the supernatants of blood components with a commercial enzyme‐linked immunosorbent assay (ELISA). The effect of leukocyte deletion and storage was evaluated by testing whole blood, buffy‐coat‐reduced, and leukocyte‐depleted red cell units. Buffy‐coat‐derived platelets and plasma were also tested. Results: HNE concentrations at day 1 were about 50 μg/l in all types of red cell components with the exception of leukocyte‐depleted red cells (<0.26 μg/l). In leukocyte‐depleted red cells, platelets and plasma, no significant increase was observed during storage. In whole‐blood units and buffy‐coat‐reduced red cells, the HNE concentrations increased steadily and often exceeded 1,000 μg/l when the units expired. Conclusion: Leukocyte depletion may limit the inadvertent infusion of bioactive substances derived from polymorphonuclear leukocytes, of which HNE is but one example. The accumulation of HNE in buffy‐coat‐reduced red cells may be greater than that of whole blood units. HNE accumulates during storage and its quantity may have pathophysiologic significance. Platelets and plasma derived from buffy coats contain some HNE, but leukocyte‐depleted red cells virtually none. However, we consider the accumulation of HNE in these components not to be clinically important. The quantities, kinetics, and occurrence in various blood components of HNE contamination differ from those observed with cytokines.
Errors in Interpreting the Pretransfusion Bedside Compatibility Test
Authors: Dujardin P.‐P. , Salmi L.R. , Ingrand P.
Published: December 2004

Background and Objective:
Analysis of reports of incidents, involving ABO incompatibility suggests that the main problem is poor interpretation of the pretransfusion bedside compatibility test (PBCT). We studied sources of error as experienced by nurses as to the blood groups of donor blood and of the recipient. Materials and Methods: According to their seniority in the profession and on the ward, 48 nurses were randomly selected from four transfusion sectors of the University Hospital of Grenoble, France. Each nurse interpreted 24 photos of PBCTs, including some with procedural irregularities, and was asked to assess the compatibility of the blood types of the donor and the recipient. At random, half the nurses were provided with a diagram to facilitate interpretation. Results: The overall frequency of errors was 39.8%. Errors were fewer when the tests were interpreted as compatible (7.3%) or incompatible (6.3%), and when the nurse had been in the profession between 3 and 5 years and in the ward less than 3 years (25.5%), or worked in hematology (34.7%) or anesthesia (36.5%). Use of the diagram limited the number of errors, provided the test was interpretable (22.2%). Conclusion: PBCTs cannot be considered a valid safety procedure. We need other, more effective methods to reduce the risk of incompatibility accidents.
Assessment of Liver Disease and Biochemical and Immunological Markers in Swedish Blood Donors with Isolated GB Virus C/Hepatitis G Virus Viremia
Authors: Björkman Per , Sundström Gunnar , Veress Béla , Widell Anders
Published: December 2004

Objective:
To investigate signs of liver disease, and biochemical and immunological markers in blood donors with isolated GBV‐C/HGV viremia. Methods: Eighteen donors with isolated GBV‐C/HGV viremia were followed up 3–5 years after initial identification. Testing for GBV‐C/HGV RNA, GBV‐C/HGV‐E2 antibodies and a range of biochemical and immunological tests was performed. Thirteen donors consented to liver biopsy. Results: Twelve donors remained GBV‐C/HGV viremic at follow‐up. Five donors had developed E2 antibodies. Liver biopsies revealed mild portal inflammatory lesions in 6/11 individuals with persistent viremia, and steatosis in 10/13 biopsied donors. Conclusion: Steatosis and mild portal inflammatory lesions were found in liver biopsies from several blood donors with isolated GBV‐C/HGV viremia.
Continuous Monitoring of Arterial Blood Gases and pH during Intraoperative Rapid Blood Administration Using a Paratrend Sensor
Authors: Vretzakis G. , Papaziogas B. , Matsaridou E. , Vasiliadou G. , Papadopoulos G. , Patsialas C. , Kostopoulou F.
Published: December 2004

Background and Objectives:
The aim of this study was to determine the effects of rapid transfusion of packed red cells on the arterial blood gases and acid‐base status of the recipient. Materials and Methods: We studied 16 patients (mean age 66.3±9.9 years) who received rapid transfusion of 632.8±287.2 g of packed red cells in CPDA‐1, stored before use for a period of 15.2±4.4 days. During transfusion, monitoring of pH, PCO2 and PO2 was continuous using an intra‐arterial multiparameter sensor (Paratrend 7, Biomedical Sensors, UK). Results: The rate of the transfusion was 73.1×9.6 g/min and the duration of observation was 35.8±12.8 min. Arterial pH decreased from 7.446±0.023 to 7.385±0.034 (p<0.001) and PCO2 increased from 32.31±1.35 to 36.41±1.86 mmHg (p<0.001). ΔpH and ΔPCO2 showed significant correlation to the weight and the age of the transfused blood (p<0.001 for both dependent variables). The rate of pH change was positively but insignificantly correlated to the rate of the transfusion. Base excess was significantly decreased and end‐tidal CO2 (PetCO2) was increased from 25.8±2.0 to 28.1±2.3 mmHg (p<0.05), significantly correlating to the amount and age of the administered component (p<0.05). PetCO2 was not elevated when PCO2 changes were minimal. Alterations in PO2 were not specific and our clinical impression was that they were related to unmeasured parameters.Conclusion: Our findings suggest that the fall in pH and the elevation in PCO2 which occur during rapid transfusion of packed red cells may go undetected or be misinterpreted if the acid‐base status of the recipient is not monitored continously. These alterations are mainly of metabolic character and depend on the amount and age of the transfused component. Our data suggest that arterial sampling is essential during massive transfusions.
Storage of Platelets in Additive Solutions: Effects of Phosphate
Authors: Gulliksson H. , Larsson S. , Kumlien G. , Shanwell A.
Published: December 2004

Background and Objectives:
In a previous study, low adenine nucleotide levels and a reduced rate of glycolysis were found in platelet concentrates (PCs) prepared by apheresis and stored in a platelet additive solution (PAS). Our objective was to investigate whether the use of PAS with or without phosphate can influence platelet metabolism in a similar way. Materials and Methods: The in vitro effects of storage in either plasma or a PAS (T‐Sol or PAS‐III, both containing citrate, acetate and sodium chloride, PAS‐III containing also phosphate) of buffy‐coat‐de rived pooled platelet concentrates (BC PCs) and apheresis platelets were investigated. The use of PAS implies inclusion of some plasma (20 or 35%). Paired studies over 7 days included investigation of cell counts, pH, PO2, PCO2, bicarbonate, glucose, lactate, adenine nucleotides, and extracellular adenylate kinase activity as a marker for disintegration of platelets. The expected concentration of phosphate in T‐Sol is 0.6–1.8 mmol/I (with CPD plasma) and 0.2–0.6 mmol/l (with ACD plasma), and in PAS‐III, 15–25 mmol/I (calculated values). Results: BC‐PCs were compared during storage in 35% CPD plasma and 65% PAS (T‐Sol or PAS‐III) (experiment 1), or alternatively 20% CPD plasma and 80% PAS (T‐Sol or PAS‐III) (experiment 3). In both studies, PAS‐III shows similar and significantly higher rates of glycolysis in terms of consumption of glucose (0.06 vs. 0.04 mmol/day/1011 platelets) and production of lactate (0.11 vs. 0.07 mmol/day/1011 platelets) compared with T Sol. Levels of pH and adenine nucleotides were similar when 35% plasma was used. With only 20% plasma, significantly higher levels of adenine nucleotides were found with PAS‐III compared to T‐Sol. The storage of apheresis platelets in 35% ACD plasma and 65% PAS (either, T‐Sol or PAS‐III (experiment 5) gave significantly higher values for PAS‐III compared to T‐Sol with regard to consumption of glucose (0.08 vs. 0.06 mmol/day/1011 platelets), production of lactate (0.14 vs. 0.11 mmol/day/1011 platelets) and adenine nucleotide levels. Conclusion: With respect to apheresis PCs stored in media containing ACD plasma, our results suggest that the differences found are related to the concentration of phosphate. The results for BC‐PCs stored in media containing CPD plasma suggest that PAS‐III is preferable to T‐Sol as the PAS at plasma concentrations below 35%. The mechanism behind the phenomena observed with BC‐PCs is not known.
Increased Withdrawal Volume per Deposit for Pre‐Operative Autologous Blood Donation in Adolescents
Authors: Erb T. , Möller R. , Christen P. , Signer E. , Frei F.J.
Published: December 2004

Objectives:
This study was undertaken to assess the feasibility, tolerance, haemodynamic and haematologic effects of an aggressive phlebotomy schedule for autologous blood donation (ABD) in adolescents undergoing major orthopaedic surgery. Methods: Twenty adolescents were studied prospectively; 10 patients in group A donated 20% of the circulating blood volume on 2 occasions, whereas 10 patients in group B donated 10% on 4 occasions. Results: The amount of blood donated, subjective tolerance, cardiovascular changes during the procedure and pre‐operative haemoglobin level did not differ between the study groups (group A 111±16 vs. group B 106±10 g/l). The increase in erythropoietin was greater and occurred sooner in group A than in group B. Conclusion: In adolescents ABD is feasible with a reduced number of appointments as they demonstrate tolerance to phlebotomies with a volume which is double the standard per deposit.
Press Review
Authors: Crisa L. , Circulli V. , Smith K.A.
Published: December 2004

Human Cord Blood Progenitors Sustain Thymic T‐Cell Development and a Novel Form of Angionensi