bluuebunny/crossref_metadata_2025 huggingface dataset:
Fine Characterization of a Series of New Monoclonal Antibodies Directed against Glycophorin A
Authors: Rasamoelisolo M. , Czerwinski M. , Bruneau V. , Lisowska E. , Blanchard D.
Published: December 2004

Objectives: Glycophorins A (GPA) and B (GPB) are the major sialoglycoproteins of the human erythrocyte (RBC) membrane. To prepare tools for the analysis of GPA and GPB, we produced a series of new monoclonal antibodies (mAbs) that identified epitopes of GPA. Methods: Seven murine monoclonal antibodies directed to glycophorin A (GPA) were fully characterized by agglutination of untreated and enzyme‐treated human erythrocytes, inhibition of agglutination using chemically modified glycophorins and peptides from GPA, immunoblotting, and binding to synthetic peptides on plastic pins. Results: The antibodies identify epitopes located on four different portions of GPA: (1) NaM13‐6D2 binds to the N‐terminal portion of GPA and GPB carrying the N blood group antigen; (2) NaM26‐3F4 recognizes the homologous portion of GPA and GPB corresponding to their amino acids 6‐26; (3) NaM10‐2H12, NaM16‐IB10 and NaM10‐6G4 are specific for the amino acid sequence 38‐45 of GPA; and (4) NaM37‐5F4 and NaM13‐4E4 bind to the amino acid residues 119‐124 located on the intracellular ponion of GPA. Conclusion: These antibodies represent precise tools to investigate GPA and related molecules in different cells and tissues.
Effectiveness of Leukocyte Filters in Reducing Tumor Cell Contamination after Intraoperative Blood Salvage in Lung Cancer Patients
Authors: Perseghin P. , Viganò M. , Rocco G. , Della Pona C. , Buscemi A. , Rizzi A.
Published: December 2004

Objectives: In an effort to reduce allogeneic blood transfusions in patients undergoing elective surgery for lung cancer, we investigated the effectiveness of a method of processing shed blood with an automated device for intra operative blood salvage (IOBS) and filtration with a 3rd‐generation polyester filter to remove tumor cells. Methods: Sixteen patients were operated on for different types of lung cancer. We searched for malignant cells in pre‐ and postprocessed shed blood employing density gradient centrifugation, staining of cytospins with hematoxylin‐eosin, and antibodies to human cytokeratins. Results: In 9 out of 16 cases (56%), neoplastic cells were detected in prefiltration samples, but none were found in postfiltration cytospins. Conclusion: IOBS combined with appropriate filtration could be a very useful and safe tool in reducing allogeneic blood transfusion in cancer patients.
Citrate‐Dependent Auto‐Antibody Causing Error in Blood Grouping
Authors: Joshi S. R.
Published: December 2004

Background and objectives: Errors in blood grouping are occasionally due to anomalous antibodies or extraneous materials in the reagents. We investigated the case of a volunteer blood donor who was originally grouped as AB, Rh‐positive, by slide grouping. However, her washed red cells showed group B, which agreed with her reverse serum grouping. Materials and methods: Standard serologic techniques were employed throughout. Results: Investigation revealed an unusual autoantibody in the donor's blood that preferentially agglutinated the red cells in the presence of citrate. Because the grouping reagents contained citrate, this led to a false grouping when whole blood was tested rather than washed red cells. The antibody reacted at 37 °C and at 4 °C, but failed to agglutinate red cells when 0.1 M solution of monosaccharides was added to the reaction mixture. The antibody agglutinated all red cells except 6 examples of the ‘Bombay’ phenotype included in the panel. The antibody was neutralized by H secretor saliva, and its activity was abolished by DTT reagent, indicating its possible IgM nature. Conclusion: These findings suggest that this anomalous antibody has anti‐H specificity.
Molecular Basis of Kell Blood Group Phenotypes
Authors: Lee Soohee
Published: December 2004

The molecular basis of different Kell blood group phenotypes is reviewed. Sequence analysis of the Kell gene (KEL) established that single base substitutions, resulting in amino acid changes, are responsible for the different phenotypes. Most of the amino acid substitutions, with the exception of the one responsible for expression of KEL6 (Jsa), occur at the amino‐terminal half of the protein, a domain that has least amino acid homology with a family of zinc endopeptidases, which include neutral endopeptidase 24.11 and endothelin‐converting enzyme‐1. Some of the genes were expressed in transfected cells and typed with alloantibodies to confirm that the identified mutations are responsible for antigen expression. Clinical applications of Kell blood group genotyping which include prenatal diagnosis to monitor hemolytic disease of the newborn are discussed.
Response of the Erythron and Erythropoietin to Autologous Blood Donations in Paediatric Subjects. Is Erythropoietin Supplement Necessary?
Authors: Han P. , Stacy D.
Published: December 2004

Objectives:This study was undertaken to evaluate the need for erythropoietin (Epo) therapy to augment autologous blood collection in adolescents undergoing spinal corrective surgery. Methods:We measured serum Epo and parameters of iron metabolism in 35 adolescents undergoing autologous blood collection for orthopaedic surgery. Ages of subjects ranged from 11 to 16 years (mean 15.5 years) with a female predominance. Generally, 10% of intravascular blood volume was collected once a week up to a total of three collections. Results:There was an average 2.5‐fold rise in serum Epo over the period of blood collection. Epo increased immediately after blood collection. There was a 1.4‐fold rise in reticulocyte count, consistent with the Epo response, and an average of 1.5 units of red blood cells (200 ml/unit) being produced over this period. Despite this there was an average fall of 2 g/dl (15%) in haemoglobin level. Serum ferritin and transferrin saturation also fell. Conclusions: Paediatric subjects are able to donate the required units of blood as they have a good Epo response to mild anaemia. The amount of blood donated did not exceed their total mobilisable iron and the iron supplement was adequate for red cell synthesis.
Markers for Transfusion‐Transmissible Infections in North Indian Voluntary and Replacement Blood Donors: Prevalence and Trends 1989–1996
Authors: Nanu Ambika , Sharma S.P. , Chatterjee Kabita , Jyoti P.
Published: December 2004

Background and Objectives: Twenty five to 30% of multiple‐transfusion recipients in India show evidence of infection with both HBV and non‐A non‐B hepatitis or HCV. To be licensed, blood banks must screen each donor unit for HBsAg, antibodies to HIV‐1 and ‐2, and VDRL. Materials and Methods: Between 1989 and 1996, 132,093 voluntary and replacement donors at this centre were screened for the above markers, using commercially available kits. Some 19,531 donors were screened for HCV antibodies in 1995 and 1996 with an inhouse EIA, using a synthetic peptide from core, NS3 and NS4 region proteins of all major HCV strains. Results: Data were tabulated annually. The proportion of voluntary donors increased from 15 to 32% during the eight years. HBsAg rates remained below 2.5%, antibodies to HIV increased from 0.04% to 0.55%, and VDRL reactivity increased from 0.23 to 0.52% between 1989 and 1995. Prevalence of all three infections showed a small but significant drop in 1996, and all three were significantly less frequent in voluntary donors. HCV antibodies were detected in 1.49% of donors tested. Donors with multiple infections were uncommon. Conclusions: A change to voluntary blood transfusion service and addition of anti‐HBc, anti‐HCV, and possibly HIV antigen to the mandatory list for screening donor blood would reduce posttransfusion infections. Indigenous production of these assays would mitigate the financial burden, as has been the experience with the in‐house HCV assay.
Evaluation of a Malaria Antibody ELISA and Its Value in Reducing Potential Wastage of Red Cell Donations from Blood Donors Exposed to Malaria, with a Note on a Case of Transfusion‐Transmitted Malaria
Authors: Chiodini P. L. , Hartley S. , Hewitt Patricia E. , Barbara J. A. , Lalloo K. , Bligh J. , Voller A.
Published: December 2004

Background and objectives: Blood donations are often wasted for lack of a satisfactory procedure to evaluate donors potentially exposed to malaria. Materials and methods: We evaluated a commercial ELISA for the detection of antibodies to malaria and compared it with an immunofluorescent antibody test (IFAT). Results: When 5,311 sera from routine non‐exposed donors were tested, 24 (0.45%) were positive by the ELISA, using a Plasmodium falciparum antigen. Seventeen were subjected to confirmatory testing but none were positive by IFAT. Of 1,000 donors potentially exposed in endemic areas 15 (1.5%) were repeatably reactive by ELISA. 10 of these were tested by IFAT and 2 were positive. When 150 patients attending the Hospital for Tropical Diseases in London with acute malaria were tested, 73% of those infected with P. falciparum were repeatably reactive for malarial antibodies by ELISA and 56% with Plasmodium vivax. Of 88 stored clinical sera tested by both IFAT and ELISA 56 were positive by IFAT and of these 52 (93°/0) were positive by ELISA. Conclusion: The ELISA is sufficiently sensitive and specific to screen at‐risk donors. Its use could safely retrieve 40,000 red cell units currently discarded each year in Great Britain.
Use of SERVQUAL to assess Clinicians' Satisfaction with the Blood Transfusion Service
Authors: Raspollini E. , Pappalettera M. , Riccardi D. , Parravicini A. , Sestili S. , Rebulla P. , Sirchia G.
Published: December 2004

Background and objectives: Limited information is available on the level of satisfaction of clinicians with services delivered by blood banks. The purpose of this study was to evaluate the satisfaction of clinicians with our blood transfusion service. Materials and methods:We prepared a questionnaire based on SERVQUAL, a method used to measure customers' appreciation of quality of service, by assessing the gap between perceived and expected quality. The questionnaire consisted of 14 items grouped according to five dimensions of quality of service: assurance, empathy, responsiveness, reliability, tangibles. Clinicians were asked to give two scores on a scale from 1 to 7 for each item, score (e) representing what they expected from an ‘excellent' service, score (r) how they graded the service received. We considered wide differences in scores of service expectation and receipt for a question to be indicative of either service above expected levels (r>e) or service below expectation (r<e); similar scores for both expected and received service (within 1 point on the grading scale) were taken to indicate that the service received was that which was expected. Results: A total of 184 questionnaires (49%) were returned. For the 14 items considered, the proportion of clinicians expressing levels of satisfaction similar to or above expectation ranged from 67 to 96%. Three critical areas, which clinicians considered important (expectation scores 6–7) were associated with satisfaction below expectation in more than 20% of responders. They were: clarity of procedures, clarity of blood request forms, and convenience of blood request and issuing times, which were rated as important by 77, 80 and 72% of clinicians, respectively. Conclusion: SERVQUAL was useful to gather information on the level of clinicians' satisfaction with our transfusion service.
Influence of Autologous Blood Transfusion on Natural Killer and Lymphokine‐Activated Killer Cell Activities in Cancer Surgery
Authors: Heiss M. M. , Fasol‐Merten K. , Allgayer H. , Ströhlein M. A. , Tarabichi A. , Wallner S. , Eissner H. I. , Jauch K.W. , Schildberg F.W.
Published: December 2004

Background and objectives: Immunosuppression associated with blood transfusion may influence postoperative infection rates. It may also affect the prognosis of patients treated surgically for colorectal cancer. To control this effect, study protocols have applied autologous blood donation programs, which are thought to be immunologically neutral. However, evidence has emerged that blood donation itself might have suppressive effects on natural killer (NK) cell activities. At present, there are no data available on the effects of autologous blood transfusion on NK or lymphokine‐activated killer (LAK) cells. This might be of interest as LAK cells may be active in tumor control. Materials and methods: 26 patients who underwent surgical resection for colorectal cancer, were assigned at random into two groups: (1) autologous blood donation and transfusion, or (2) allogeneic blood transfusion. NK and LAK activities were determined before blood donation, at surgery, and on the 3rd and 8th postoperative day. Results: Blood donation induced a small decrease in NK and LAK activities. The postoperative courses of the two groups differed. In the allogeneic group, NK activity (−50%, p = 0.018) and LAK activity decreased (−60.7%, p = 0.043), whereas in the autologous group the decline in LAK was less pronounced (−33.7%, p = 0.091), and their NK activity even increased (+17.4%, p = 0.315). NK activity was modulated differently in the two study groups (0.0036). Differences in LAK activities were found between the 3rd and 8th day postoperatively (p = 0.354). Conclusions: In patients receiving autologous blood transfusion, postoperative suppressed NK and LAK activities were modulated. This implies that autologous blood transfusion is not immunologically neutral, but has an intrinsic immunomodulatory potential.
How to Stabilize the Level of lonized Calcium and Citrate during Plateletpheresis
Authors: Farrokhi P. , Farahmand H. , Bismuth A. , Suarez C. , Ducot B. , Gillon M. C. , Debat P. , Laskowski P. , Pinon F.
Published: December 2004

Background and objectives: Citrate reactions are uncomfortable and potentially dangerous to apheresis donors. Reduction of citrate increases comfort, but may lead to platelet clumping. Materials and methods: We describe a protocol for stepwise reduction of the volume of ACD‐A injected during plateletpheresis. This protocol has been carried out in 45 healthy donors with the Cobe 2997 (Cobe) cell separator, and in 35 with the Fenwal‐CS 3000 (CS). Results: Using this protocol, during the first hour of platelet collection, ionised calcium decreased on average by 18% for CS and by 18.4% for Cobe. During the second hour, Ca2+ and citrate ion concentration did not change with either Cobe or CS (about 65% of citrate ion load is eliminated). We observed mild signs of neuromuscular hyperexcitability in only 22% and 28% of donors with Cobe and CS, respectively. We also found a significant reduction of phosphate ions (p < 0.0001) at the end of the procedures. Conclusions: With this stepdown citrate reduction protocol, we obtained a significant reduction of injected citrate without the complication of platelet clumps.
DNA Recommendations 1997 of the International Society for Forensic Genetics
Authors: Olaisen B. , Bär W. , Brinkmann B. , Budowle B. , Carracedo A. , Gill P. , Lincoln P. , Mayr W.R. , Rand S.
Published: December 2004

The DNA commission of the International Society for Forensic Haemogenetics (ISFH) has over the years published a series of documents providing guidelines and recommendations concerning the application of DNA polymorphisms to problems of identification. This latest report provides recommendations relating to the nomenclature of short tandem repeat system (STR) typing systems which are at the forefront of systems used at present by forensic scientists and are likely to remain so for the immediate future.
Improved Blood Preservation with 0.5CPD Erythro‐Sol. Coagulation Factor VIII Activity and Erythrocyte Quality after Delayed Separation of Blood
Authors: Solheim B. G. , Bergerud U. E. , Kjeldsen‐Kragh J. , Brosstad F. , Mollnes T. E. , Högman C. F. , Eriksson L. , Schutz R.
Published: December 2004

Background and objectives: Delay between blood collection and the separation of its components may result in lowered yield of factor VIII (FVIII) and loss of 2,3‐biphosphoglycerate (2,3‐BPG). This study was to see whether the use of 0.5 CPD resulted in better preservation of FVIII and maintenance of 2,3‐BPG. Materials and methods: 55 units of blood were collected in 0.5CPD and 48 in CPD SAG‐M. Ten of the collections were paired, so that the same donors were bled a single session partly in an 0.5 CPD system and partly in CPD SAG‐M. After collection, the blood was promptly cooled to 20°C and stored at that temperature for up to 24 h. Results: Preservation of FVIII activity was significantly better in 0.5CPD compared with CPD. The content of von Willebrand factor was stable in the anticoagulant solutions for 24 h at that temperature. Plasma separated from both media had low levels of prothrombin fragment 1 + 2 and complement activation. Paired collections substantiated previous reports that red cell storage is significantly improved in 0.5CPD compared with CPD SAG‐M with respect to 2,3‐BPG and haemolysis. Conclusions: Red cell metabolism and oxygen‐releasing capacity are kept at acceptable levels in 0.5CPD blood for 24 h at 20°C before component separation. The concentration of red cell 2,3‐BPG remained at normal or slightly subnormal levels during further storage in 0.5CPD at 4°C for 2–4 weeks before gradual decay to an average of 39% at 48 days.
Immunomodulating Effect of Blood Transfusion: Is Storage Time Important?
Authors: Mynster Tommie , Dybkjœr Esben , Kronborg Gitte , Nielsen Hans Jørgen
Published: December 2004

Objectives: TNF‐α and IL‐2 are important cytokines in macrophage and T‐lymphocyte activity against infection and dissemination of malignant cells. We studied the influence of supernatants from stored whole blood and buffy‐coat‐depleted SAGM (saline, adenine, glucose and mannitol) blood in stimulating TNF‐α and IL‐2 release in an ex vivo assay. Methods: Supernatants of 10 units of whole blood and 10 units of SAGM blood were collected after 1, 21 and 35 days of standard blood bank storage. Heparinized blood from 20 healthy volunteers (as ‘recipients’), corresponding in ABO and Rh type to the stored blood, were used in a culture system with LPS and PHA as stimulators of TNF‐α and IL‐2 release. The effect of added supernatants, from either stored whole blood or SAGM blood, on cytokine release was evaluated compared to saline as control. TNF‐α concentration was analyzed by ELISA after culture for 24 h and IL‐2 after 72 h, respectively. Results: Supernatants from both stored whole blood and SAGM blood showed a significant decrease in both LPS‐ and PHA‐stimulated TNF‐α release that was dependent on storage time. IL‐2 was not detected in response to LPS stimulation. PHA‐stimulated IL‐2 release was significantly reduced and related to storage time of both whole blood and SAGM blood. Conclusions: Recipient cytokine release induced by blood transfusion seems to be dependent on storage time. This may have implications in transfusion‐induced immune modulation.
Neonatal Alloimmune Thrombocytopenia due to Anti‐HPA‐1b (PLA2) (Zwb)
Authors: Winters J.L. , Jennings C.D. , Desai N.S. , Dickson L.G. , Ford R.F.
Published: December 2004

Background: Neonatal alloimmune thrombocytopenia is a rare condition due to passively acquired maternal antibodies directed against paternal platelet antigens inherited by the infant. Only 5 cases have been reported due to antibodies against HPA‐lb (PLA2) (Zwb). Case Report: We report a case of neonatal alloimmune thrombocytopenia due to anti‐HPA‐lb in the second pregnancy of a 26‐year‐old Caucasian female. The male infant was treated with a 5‐day course of intravenous immunoglobulin without complications. We report the HLA phenotype of the infant's mother and summarize the previous case reports due to anti‐HPA‐lb. Conclusion: Based on this case and a review of the literature, intravenous immunoglobulin as well as random donor exchange transfusion and random donor platelet transfusions are effective in the treatment of neonatal alloimmune thrombocytopenia due to anti‐HPA‐lb. Obvious associations between HLA alleles and sensitization to HPA‐lb have not been elucidated.
Human Parvovirus B19: Prevalence of Viral DNA in Volunteer Blood Donors and Clinical Outcomes of Transfusion Recipients
Authors: Jordan J. , Tiangco B. , Kiss J. , Koch W.
Published: December 2004

Background and Objectives: Blood donor units are not screened for human parvovirus B19 (B19) even though it can be acquired via blood products. We estimated the prevalence of B19 in a US volunteer blood donor population and determined the clinical outcomes of transfusion recipients. Materials and Methods: Donor units were screened for B19 DNA by PCR, and positive units analyzed by EIA for B19 Ig. Unit usage was determined and recipient chart review conducted. Results: B19 DNA was detected in 11/9, 568 allogeneic units (0.1%), of which 3 had no measurable B19 Ig. One individual developed anemia consistent with B19 infection after receiving a DNA+ unit lacking B19 Ig. Conclusions: The apparent low incidence of disease in patients transfused with B19 DNA+ components may be due to coexistence of neutralizing antibodies in donors and/or recipients.
Does Hyperconcentration Result in Platelet Activation?
Authors: Gollehon T.J. , King D.E. , Craig F.E.
Published: December 2004

Background and Objectives: Hyperconcentration of platelets may lead to platelet activation and loss of platelet function. Materials and Methods: Platelet activation following hyperconcentration was assessed using flow‐cytometric detection of platelet P‐selectin expression and platelet swirling. Results: Platelet hyperconcentration led to a minimal increase in P‐selectin expression and no differnce in platelet swirling. Conclusion: Hyperconcentration was not associated with a clinically significant change in platelet activation and had no significant effect on platelet quality as detected by pH and platelet yield.
A Flow‐Cytometric Equivalent of the Kleihauer Test
Authors: Nelson M. , Zarkos K. , Popp H. , Gibson J.
Published: December 2004

Background and Objectives: The Kleihauer slide test is in general use to screen obstetric patients for possible fetomaternal haemorrhage. Since 1993, Rh(D)‐negative patients have been tested in our laboratory by a flow‐cytometric method detecting Rh(D)‐positive fetal cells, a method which offers improved sensitivity and accuracy. We report another flow‐cytometric method of broader application which quantitates cells according to haemoglobin F (HbF) content. Materials and Methods: The red cells are fixed with glutaraldehyde and permeabilized by exposure to Triton X‐100. A polyclonal sheep antibody to HbF is incubated with the cells followed by a fluorescein‐labelled anti‐sheep antibody. Results: Quantitation of the percentage of fetal cells following a FMH can be achieved irrespective of the blood groups of either mother or infant, and the presence of maternal F cells need not interfere since the intensity of staining is usually less than that of fetal cells. Two of 19 transfusion‐dependent patients with β‐thalassaemia have been found to have red cells indistinguishable from fetal cells on the basis of HbF content, but these patients also have been found to give positive results by the Kleihauer test. Conclusion The flow‐cytometric method may serve to replace the traditional Kleihauer test since it appears to offer improved accuracy and objectivity.
A Method for the Quantitative Assessment of Platelet‐Induced Clot Retraction and Clot Strength in Fresh and Stored Platelets
Authors: Reid T. J. , Snider R. , Hartman K. , Greilich P. E. , Carr M. E. , Alving B. M.
Published: December 2004

Background and Objectives: The changes that occur in platelets as they undergo storage have been documented by aggregometry as well as by flow cytometry. However, one of the most essential platelet functions, the induction of clot retraction, has not been quantitatively assessed in stored platelets. We describe two potentially useful methods, platelet‐induced clot retraction and clot strength, to assess effect of storage of platelets in blood banks or of platelet preparations subjected to freezing or freeze‐drying. These methods have previously been developed for bedside monitoring of patients receiving c7E3 (Reopro®). Materials and Methods: Platelet‐induced clot retraction (PICR) and clot strength were measured with the Hemodyne ™ and Thromboelastograph®, respectively. Paired Study: Fresh platelet concentrates (n = 3) were obtained from leukapheresis donors and divided into two equal units; one unit was tested within 4 h of collection and the other stored for 5 days at 22°C in a platelet incubator and tested. Unpaired Study: Fresh platelet concentrates (n = 15) were obtained from leukapheresis donors and tested within 4 h of collection and compared to outdated platelets (n = 30; random or single donor) that had been stored for 5 days at 22°C in a platelet incubator. Alternative Presevation Methods: Lyophilized platelets, platelets chilled to 4°C, platelets frozen at −70°C in 5% dimethyl sulfoxide (DMSO) or in the absence of a cryoprotectant Results: Paired Study: Stored platelets demonstrated an increase in PICR; the difference was not significant (p = 0.55). There was no difference in clot strength between fresh and outdated platelets (p = 0.90). Unpaired Study: When compared to fresh platelets, stored platelets demonstrated a 2‐fold higher PICR (p = 0.0011). On the other hand, there was no difference in the time to onset of PICR (p = 0.08) and there was no difference in clot strength between fresh and outdated platelets (p = 0.14) Alternate Preservation Methods: In contrast, PICR and clot strength were reduced in platelets frozen at −70°C in 5% DMSO and absent in lyophilized platelets, in platelets frozen at −70°C in the absence of cryoprotectants or stored at 4°C. Conclusion: The data indicate that the ability of platelets to indcue clot retraction and to enhance clot strength is not altered by storage, despite functional abnormalities in aggregation and agglutination. These data suggest that quantitative measurements of PICR and clot strength may be simple, useful tools for assessing the function of stored platelet concentrates, platelets that have undergone freezing or exposure to alternative buffers and for evaluating platelet functions relevant to PICR.
Anti‐Rh D Activity of Commercial Intravenous Immunoglobulin Preparations
Authors: Turner C.E. , Thorpe S.J. , Brasher M.D.R. , Thorpe R.
Published: December 2004

Background and Objectives:
The presence of antibodies against Rh D in intravenous immunoglobulin (IVIG) products has been proposed as causing adverse reactions and blood grouping problems, although data on the incidence and titre of such antibodies is sketchy. This study was conducted to investigate methodology for carrying out haemagglutination tests on IVIG products, and to examine in retrospect 200 IVIG batches for the presence of anti‐D and other haemagglutinins. Materials and Methods: Batches of commercial IVIG products were tested using the low‐ionic‐strength indirect antiglobulin test (LIAT). Positive batches were futher investigated using a typed red cell panel and direct haemagglutination tests. Results: 44 batches of IVIG gave positive LIAT. Of these, 12 contained specific anti‐D. 11 batches had LIAT anti‐D titres of 2–8; a further batch contained anti‐D with a LIAT titre of 64. The remaining 32 batches agglutinated all erythrocyte phenotypes tested with LIAT titres ≤4; 14 of these batches showed positive direct haemagglutination and the presence of rouleaux. Conclusion: 6% of commercial IVIG batches contained specific anti‐D, including one batch (0.5%) showing unusually high titre. 16% of batches contained haemagglutinins of unknown/non‐specificity.
Transfusion‐Associated Graft‐versus‐Host Disease in an Immunocompetent Patient following Cardiac Surgery
Authors: Rososhansky Sarah , Badonnel Marie‐Claude H. , Hiestand Lauren L. , Popovsky Mark A. , Szymanski Irma O.
Published: December 2004

Objectives:
We determined which of the 22 blood components obtained from unrelated donors and transfused to an apparently immunocompetent patient following open heart surgery caused transfusion‐associated graftversus‐host disease (TA‐GVHD). Methods: Serologic and molecular methods were used to type the donors, the patient's family members, and the patient's postmortem tissues for HLA and a genetic marker on chromosome 17. Results: Two donors were homozygous for the HLA class I antigens A1 B8, for which the patient was heterozygous. Both donors were heterozygous, not homozygous as expected, for the class II alleles. One of them had the same class II alleles as the patient (DRB1*0301, DRB3*0101/DRB1*0404, DRB4*0103). The patient's tissues were chimeric for restriction fragments at 17 p 13 of this donor. Conclusion: One‐way HLA match leading to TA‐GVHD can be caused by donor blood that is homozygous for class I and heterozygous for class II alleles. Two blood components given to our patient had such one‐way HLA match. Class II alleles of the lymphocytes in one component were identical with those of the recipient and caused TA‐GVHD. Class II alleles of the lymphocytes in the other component differed from those of the recipient and were eliminated either by the immune system of the patient or the lymphocytes that caused the TA‐GVHD (graft versus graft).
Hepatitis E Virus: Relevance in Blood Donors and Risk Groups
Authors: Mateos María Luisa , Camarero Cristina , Lasa Elena , Teruel José Luis , Mir Nuria , Baquero Fernando
Published: December 2004

Background and Objectives:
Hepatitis E virus (HEV) infection usually causes acute self‐limited disease. HEV is associated with faecal‐contaminated drinking water, but other Vectors, such as blood, are possible. The aim of this study was to investigate the prevalence of HEV in blood donors and in two groups at high risk for parenteral infections, namely, haemodialysis patients, and children infected with HCV via blood transfusion. Materials and Methods: We investigated the prevalence of anti‐HEV in 863 blood donors, 63 haemodialysis patients, and 42 children infected post transfusion with HCV. Results: The prevalence rates were 2.8, 6.3%, and zero, respectively. Conclusions: (1) The incidence of HEV in Spain is similar to that in other Western European countries, and (2) HEV is probably not transmitted parenteally to children.
GB Virus Type C Viremia and Envelope Antibodies among Population Subsets in The Netherlands
Authors: Jongerius J.M. , Boland G.J. , van der Poel C.L. , Rasch M.C. , Italiaander E. , van der Reijden J.J. , Friedman P.N. , Cockerill J.J. , van Leeuwen E.F. , van Hattum J.
Published: December 2004

Background and Objectives:
Two new flaviviruses, hepatitis G virus and GB virus type C (GBV‐C), are possibele causative agents for non‐A‐E hepatitis. In this study we established the prevalence of GBV‐C markers in various population subsets in The Netherlands by assays for GBV‐C antibodies and GBV‐C nucleic acid. Materials and Methods: We tested specimens from groups of patients with hepatitis of various causes, intravenous drug users (IVDUs), and blood donors for GBV‐C BNA (LCx® GBV‐C assay, Abbott Laboratories), and for antibodies to the GBV‐C envelope E2 protein (GBV‐C anti‐E2) with an enzyme immunoassay (Abbott Laboratories). Patients and donors were represented in one group only. Results: GBV‐C RNA and GBV‐C anti‐E2 prevalence were, respectively, 2/34 (6%) and 3/34 (9%) among patients with non‐A‐E hepatitis, 2/10 (20%) and 0/10 (0%) among hepatitis B virus patients, 10/40 (25%) and 19/40 (48%) among hepatitis C virus (HCV) patients, 1/8 (13%) and 0/8 (0%) among patients with autoimmune hepatitis (AIH), 24/102 (24%) and 72/102 (71%) among IVDUs, 1/34 (3%) and 2/34 (6%) among blood donors with indeterminate anti‐HCV recombinant immunoblot assay reactivity, and 3/250 (1.2%) and 8/250 (3.2%) among first‐time blood donors. the profile of simultaneous GBV‐C RNA positivity plus GBV‐C anti‐E2 positivity was found in 2/40 (5%) HCV patients, 4/102 (4%) IVDUs, and 1/250 (0.4%) first time blood donors. Conclusion: GBV‐C infection appears not to be a major cause of non‐A‐E hepatitis and AIH, but is associated with parenteral risk. The prevalence of GBV‐C viremia in first time blood donors is higher than that of HCV (1.2 vs. 0.04%), but, GBV‐C viremia in IVDUs is lower than HCV (24 vs. 59%). Most IVDUs have probably previously been exposed to GBV‐C given the very high prevalence of GBV‐C anti‐E2 (71%). Most persons with GBV‐C markers are GBV‐C marker are GBV‐C RNA‐negative and anti‐E2 confirmed positive, suggesting that GBV‐C infection is transient.
Automated Separation of Whole Blood in Top and Bottom Bags into Components Using the Compomat G4
Authors: van der Meer P.F. , Pietersz R.N.I. , Hinloopen B. , Dekker W.J.A. , Reesink H.W.
Published: December 2004

Background and Objectives:
Whole blood can be separated by hard spin centrifugation into layers of blood components according to their specific gravity. The aim was to develop a program for an automatic separator to subsequently express the various components into their respective satellite bags in top and bottom systems with the following requirements; a red cell concentrate with a low leukocyte and platelet contamination, a ‘cell‐free’ plasma, and a buffy coat with a volume of about 50 ml with an acceptable loss of red cells. Materials and Methods: The Compomat G4 possesses an independently moving upper and lower press, to automatically express plasma or red cells to satellite bags of top and bottom systems. The influence of the extension of the lower press was studied by pooling and dividing two units of whole blood, and separating these units after centrifugation (2,960 g, 10 min) either with a program where the lower press was completely extended (program C), or with a program that left approximately 1 mm between the door and the lower press (program D). Results: The program (program d), where the lower press was not completely extended, yieded a buffy coat with a volume of 52±1 ml (mean ± SD, n = 36), which contained >75% leukocytes and >90% platelets of the original whole blood unit, with a red cell loss in the buffy coat of 21±1 ml (10.8±0.8% of the original volume). The leukocyte content of the red cell concentrates was 775±379 × 106 per unit, whereas the plasma contained 3±3×106 leukocytes and 4±3 × 109 platelets per unit. The pooling experiment indicated that complete extension of the lower press (program C) resulted in a significantly higher leukocyte contamination of the red cell concentrate (788±431 × 106 vs. 658±419 × 106; n = 12; p = 0.03), while there was no difference in the yield of red cells or plasma. The buffy coat produced with program D contained significantly more leukocytes (2,242±396 × 106 vs. 2,065±327 × 106, p × 0.005) and more platelets (96±14 × 109 vs. 92±17 × 109, p 0.02) per unit than with program C, probably because buffy coat cells sticking to the container wall are not expressed to the red cell concentrate, and thus remain in the buffy coat bag. Therefore, program D met our specifications for blood products. Conclusions: The Compomat G4 warrants reproducible separation of whole blood in top and bottom bags into red cells, buffy coat and plasma meeting our specifications.
Cytokine Generation in Whole Blood, Leukocyte‐Depleted and Temporarily Warmed Red Blood Cell Concentrates
Authors: Weisbach V. , Wanke C. , Zingsem J. , Zimmermann R. , Eckstein R.
Published: December 2004

Background and Objectives:
It has been suggested that inflammatory cytokines such as Interleukin (IL)‐1β, IL‐6 tumor necrosis factor‐α (TNF‐α) and IL‐8 might be responsible for a large number of non‐antibody‐mediated adverse reactions to the transfusion of blood components, especially of platelet concentrates (PCs). The aim of this study was to compare the levels of proinflammatory cytokines in different blood components containing red cells such as buffy‐coat‐free packed red cells (RBCs), filtered RBCs and whole blood (WB) during storage under several conditions. Materials and Methods: WB (CPD‐A1, n = 16) was stored for 35 days at 2–6°C; sameles were taken on days 0, 21 and 35. Buffy‐coat‐poor RBCs in additive solution PAGGS‐M (n = 16) were divided into halves, one half was leukocyte (WBC)‐depleted by filtration on day 0, both halves were stored for 49 days at 2–6°C (samples: days 0, 21, 40). furthermore, buffy‐coat‐poor, unfiltered SAG‐M RBCs (n = 16) were halved immediately after production and stored at 2–6°C until day 42 (samples: days 0, 21, 42). One half remained at room temperature for 24 h on day 3. Cytokine levels were determined with commerical enzyme‐linked immunosorbent assays. Results: Levels of IL‐1β and TNF‐α rose during storage of WB and RBCs. IL‐6 could be detected markedly above the detection threshold in WB only. At the end of storage, we detected IL‐8 in 1 of 16 units of WB tested, in 10 to 16 standard PAGGS‐M RBCs and in 15 of 16 temporarily warmed SAG‐M RBCs. Prestorage filtration of RBCs prevented the accumulation of IL‐1β and TNF‐α. Temporarily warming of RBCs for 24 h did not cause any substantial increase in cytokine levels other than IL‐8. RBCs stored in different additive solutions (PAGGS‐M versus SAGM) showed similar cytokine concentration during storage. The cytokine content of WB was very similar to that of buffy‐coat‐poor RBCs. Conclusion: Cytokine levels measured in WB and buffy‐coat‐poor RBCs result in levels which are unlikely to cause febrile reactions even in the case of massive transfusion. We conclude that, according to present knowledge, there is no reason for prestorage filtration of buffy‐coat‐poor RBCs or WB to avoid febrile transfusion reactions due to cytokine accumulation during storage.
A Newly Identified Nonsecretor Allele of the Human Histo‐Blood Group α(1,2) Fucosyltransferase Gene (FUT2)
Authors: Yu Lung‐Chin , Lee Hui‐Lin , Chu Chen‐Chung , Broadberry Richard E. , Lin Marie
Published: December 2004

Background and Objectives:
The human Secreα(1,2) fucosyltransferase gene determines the ABH secretor status and influences the Lewis phenotype of an individual. Studies were carried out on the Lewis 〈a+b‐〉 nonsecretors of different groups indigenous to Taiwan to demonstrate their se genotypes. Meth: The Lewis phenotype of the blood samples was determined by a microplate method. The se genotypes of the individuals with the Lewis 〈a+b‐〉 phenotype were analyzed by a polymerase chain reaction restriction fragment length polymorphism (PCR‐RFLP) method designed for the se alleles reported previously. PCR and cloning techniques were used to determine the coding sequence of the novel se gene. Results: A new se allele, se685, with a three‐nucleotide deletion of GTGGT to GT in the coding region of nucleotides 685 through 689 was identified in a Le 〈a+b‐〉 nonsecretor from the Ami tribe indigenous to Taiwan. The deletion predicts the loss of the amino acid Val230 in the corresponding secretor enzyme's C‐terminal segment. The distribution of the se685 allele in the Ami tribe was further verified by PCR‐RFLP analysis. Conclusion: The se gene exhibits heterogeneity with some se alleles being common but others displaying a unique distribution in different ethnic populations. The newly identified se685 allele seems to exist only in the Ami tribe indigenous to Taiwan.
Pan‐Reactive Platelet Antibodies in Post‐Transfusion Purpura
Authors: Taaning Ellen , Tønnesen Finn
Published: December 2004

Background and Objectives:
The mechanism for the destruction of a patient's own platelets in post‐transfusion purpura (PTP) is unknown. In order to test the hypothesis that the destruction of autologous platelets in PTP is related to the presence of platelet antibodies with panreactivity, we investigated sera from patients with PTP. Materials and Methods:: Sera from 12 patients with PTP were investigated for platelet antibodies by platelet‐ELISA and monoclonal antibody‐specific immobilization of platelet antigen assay. Results:: During the thrombocytopenic phase, antibodies of IgG and lgM classes with pan‐specificity against platelet GPIIb‐IIIa, GPIb‐IX and GPIa‐IIa were found together with HPA alloantibodies. After recovery, the pan‐specific antibodies disappeared or the extent of reactivity diminished, whereas the IgG HPA alloantibodies persisted. Conclusion: These findings provide evidence that transient panreactive antibodies may be responsible for the autologous platelet destruction in PTP.
Transmission Rates of Hepatitis C Virus by Different Batches of a Contaminated Anti‐D Immunoglobulin Preparation
Authors: Lawlor E. , Power J. , Garson J. A. , Yap P. L. , Davidson F. , Columb G. , Smith D. , Pomeroy L. , O'Riordan J. , Simmonds P. , Tedder R. S.
Published: December 2004

Background and Objectives:
The aim of this study was to determine the hepatitis C virus (HCV) infection rate of recipients of different batches of anti‐D immunoglobulin associated with an outbreak of HCV infection which occurred in 1977 and its relationship to the polymerase chain reaction (PCR) status of the implicated batches. This study was undertaken to determine the predictive value of HCV genome detection and quantification for subsequent infection in recipients of an HCV‐contaminated anti‐D immunoglobulin product for intravenous use. Materials and Methods: Sera from recipients of anti‐D were tested by HCV enzyme immunoassay and if found positive were subsequently tested by recombinant immunoblot assay and HCV PCR in a national HCV anti‐D screening programme set up in 1994. The HCV status of 1,342 known recipients of infectious or potentially infectious batches has been compared to the amount of HCV RNA in the anti‐D batch they received so as to determine the value of PCR in the prediction of infectivity in immunoglobulin preparations. Results: It has been demonstrated that HCV‐infected plasma derived from batches of anti‐D showing levels of viral genome in excess of 104 genomes per millilitre led to infection of up to 60% of recipients. In contrast, batches with undetectable levels of HCV genome very rarely transmitted infection. Conclusions: The presence of HCV RNA in intravenous immunoglobulin preparations which have not undergone a specific viral inactivation step is a predictor of HCV infection in recipients.
Establishment of the First International Standard for Nucleic Acid Amplification Technology (NAT) Assays for HCV RNA
Authors: Saldanha J. , Lelie N. , Heath A. ,   WHO Collaborative Study Group
Published: December 2004

Background and Objectives:
The aims of this study were the establishment of a WHO International standard for HCV RNA for nucleic acid amplification technology (NAT) assays and the determination of the HCV RNA content of the candidate standard. Materials and Methods: Twenty‐two laboratories evaluated three candidate materials (two lyophilised, AA and BB, which were derived from the same source and one a liquid preparation, CC). All samples were HCV genotype 1 with a concentration of approximately 105 genome equivalents/ml. The methods used included the Roche Amplicor assay (version 1), Chiron Quantiplex (bDNA) assay, Organon Teknika NASBA assay, Transcription Mediated assay and various in‐house assays, using single or nested primers. Results: There was reasonable agreement between the overall mean NAT detectable units/ml obtained by the different assays except for some of the in‐house assays using single primers which gave substantially lower estimates. These titres were 5.0 log10 for samples AA and BB and 4.6 log10 for sample CC. Conclusions: Sample AA was accepted as the candidate standard and assigned a titre of 105 international units (IU)/ml. The International Standard consists of a batch of vials each containing 50,000 IU/vial. Preliminary studies indicated that the material is stable at +4°C and +20°C for up to 200 days.
The Relationship between New Variant Creutzfeldt‐Jakob Disease and Bovine Spongiform Encephalopathy
Authors: Knight Richard
Published: December 2004

Creutzfeldt‐Jakob disease (CJD) has been transmitted in the laboratory and also by iatrogenic accident. However, research has failed to find evidence that its most common form (sporadic CJD) is a natural infection and, in particular, that there is a causal link with scrapie. Bovine spongiform encephalopathy (BSE) probably resulted from scrapie infection in cattle food. In the wake of the BSE epidemic, a novel clinico‐pathological form of CJD has been recognized: new variant CJD (nvCJD). This paper reviews the relationship between nvCJD and BSE and presents the accumulated evidence supporting the view that nvCJD resulted from BSE contamination of human food.
Evaluation of the Use of a Platelet‐Counting Tool in Plateletpheresis
Authors: Ho T.‐F. , Yang B.‐S. , Huang Y.‐T. , Hung C.‐T.
Published: December 2004

Objective:
For years, blood transfusion centers in Taiwan have used the Quantitative Buffy Coat (QBC®) Hematology System for platelet counts on capillary blood samples in the laboratory screening of apheresis donors. The system has not been evaluated for the prediction of yields in plateletpheresis. Methods: The QBC instruments was evaluated for reproducibility of platelet counts and compared with five electronic cell counters. We also collected both capillary and venous blood from voluntary donors before donation and counted platelets, comparing the QBC system and an electronic blood cell counter (Sysmex K1000). The correlation between donors' predonation platelet counts and plateletpheresis yields was analyzed. Results: The R values for platelet counts between the QBC Hematology System and other electronic counters are lower (0.759‐0.890) than among the electronic counters (0.929‐0.973). The mean capillary platelet count and the mean venous platelet count were 241.9±50.3±103/μl and 233.2±47.9×103/μl by the QBC system, and 244.9±54.1×103/μl and 218.9±46.5×103/μl by the Sysmex K1000, respectively. Linear regression analysis showed that platelet yields correlated well with donors' predonation platelet counts using the Sysmex K1000 counter (R = 0.7770.890, p<0.001), but not with the QBC system (R = 0.326∼.755, p<0.05). Conclusion: The QBC Hematology System is not accurate enough to determine predonation platelet counts that are to be used for calculating the number of processing cycles for plateletpheresis.