bluuebunny/crossref_metadata_2025 huggingface dataset:
The MiIII phenotype among Chinese donors in Hong Kong: immunochemical and serological studies
Authors: Poole J. , King M.‐J. , Mak K. H. , Liew Y. W. , Leong S. , Chua K. M.
Published: April 2008

Summary. The incidence of the MiIII phenotype among Chinese blood donors in Hong Kong was found to be 6·28%. Eleven individuals apparently homozygous for the MiIII gene were detected by immunoblotting with monoclonal antibodies Rl.3 and R18. Rl.3 detects an identical epitope on both glycophorins A and B and Rl 8 detects a different epitope on glycophorin A. Immunoblotting with R1.3 showed an absence of bands corresponding to normal glycophorin B. Immunoblotting with R18 showed an absence of a 58 K band, which corresponds to a heterodimer of normal glycophorin B complexing with the MiIII component, found in MiIII heterozygotes. In two families with apparent MiIII homozygous individuals, both parents of the propositi had the MiIII phenotype which implies normal autosomal inheritance of the MiIII gene. In another family, only one parent had the MiIII phenotype and the presence of an Su gene is postulated to explain the immunochemical and serological findings.
The treatment of autoimmune thrombocytopaenic purpura with anti‐D immunoglobulin: similar platelet responses in homozygous and heterozygous Rh(D) positive patients
Authors: Smith N. A. , Boughton B. J.
Published: April 2008

Summary. Twenty‐one patients with autoimmune thrombocytopaenic purpura (AITP) were treated with anti‐D immunoglobulin. There was no significant difference with low and high dose anti‐D treatment in the platelet count response between homozygous and heterozygous Rh(D) positive patients. The heterogeneous responses seen in Rh(D) positive AITP patients treated with anti‐D immunoglobulin cannot therefore be explained by differences in Rh(D) phenotypes.
Platelet apheresis for extreme thrombocytosis in an 11‐year‐old girl with CML
Authors: McCarthy L. J. , Graves V. L. , Eigen H. , McGuire W. A.
Published: April 2008

Summary. This report describes the dramatic reduction in the platelet count and ablation of symptoms by consecutive platelet apheresis in a young girl with CML who presented with an extremely high platelet count of 10,320 × 109/1. Extreme thrombocytosis, although quite rare, can be symptomatic and life‐threatening in paediatric patients and should be considered an emergency. The mechanical removal of platelets by apheresis is effective therapy and should begin as soon as possible.
The effect of intravenous immunoglobulin on placental transfer of a platelet‐specific antibody: Anti‐P1A1
Authors: Morgan C. L. , Cannell G. R. , Addison R. S. , Minchinton R. M.
Published: April 2008

Summary. The isolated perfused lobule of human placenta was used as an in‐vitro model to study the effect of intravenous immunoglobulin (IVGG) on the placental transfer of a human platelet‐specific antibody (anti‐P1A1). Normal human IgG was shown to transfer from the maternal to the fetal circulation of the placental model after a lag period of 2–3 h. IVGG also transferred across the placenta but only after a longer lag period (3–4 h) than normal human IgG at the same concentration, which suggests that IVGG may contain a factor that inhibits the transfer of its own component IgG. The sensitive Western immunoblotting technique was used to demonstrate progressive transfer of anti‐P1A1 antibody to the fetal circulation after a 2–3 h lag period. When IVGG and anti‐P1A1antibody were added simultaneously to the maternal circulation, the transfer of platelet‐specific antibody was strongly inhibited by IVGG. The inhibitory effect of IVGG on anti‐P1A1 antibody transfer was consistent for three different batches of the same IVGG product (Sandoglobulin). These studies provide the first scientific data to support the use of IVGG to inhibit antiplatelet antibody transfer as part of the antenatal management of neonatal alloimmune thrombocytopenia.
Transfusion‐induced immunosuppression and red cell clearance
Authors: Donnelly P. K. , Proud G. , Shenton B. K. , Taylor R. M. R.
Published: April 2008

Summary. While blood transfusion is increasingly implicated in the aetiology of tumour recurrence, the mechanism of this effect is unclear. Cancer‐bearing patients are known to have factors in their sera which depress the function of normal lymphocytes. It is possible that blood transfusion accentuates this natural suppression. An animal model was therefore developed to study the effect of blood transfusion on humoral immunosuppressive activity and its possible relationship to red cell clearance. WAG rats given a transfusion of chromium‐labelled allogeneic but blood‐group compatible DA rat blood, developed significantly increased (P < 0·001) levels of lymphocyte suppressive factors in plasma (maximum at 7 days) which coincided with accelerated red cell clearance (t1/2= 7 days). A transfusion of syngeneic WAG blood caused only a small transient increase in plasma suppression and red cells were cleared at a normal rate (t1/2= 13 days) consistent with previous studies. However, when syngeneic WAG red cells were lysed and the red cell membranes infused there was a rapid increase in plasma suppression (P < 0·001), similar to but less prolonged than that achieved with allogeneic blood. The immunosuppressive effect of blood transfusion may result from accelerated clearance of allogeneic or damaged syngeneic red blood cells.
A murine monoclonal anti‐N, BIRMA‐N, suitable for blood grouping in an automated system
Authors: Thompson J. M. , McDonald D. F.
Published: April 2008

Summary. BIRMA‐N, a murine monoclonal anti‐N antibody of the IgGl subclass, was assessed for suitability as a blood grouping reagent on the Olympus PK7100 automated blood grouping machine. At a selected dilution and over the pH range 5·0·8·0, the antibody performed accurately in this system as confirmed by parallel manual testing of donor blood samples with human anti‐N and commercial monoclonal anti‐N reagents. These findings show BIRMA‐N to be extremely suitable for N typing blood samples in an automated system providing a convenient, objective and cost‐effective method for large scale typing of the blood donor population.
Antibody binding to blood group antigens in relation to temperature: scanning electron microscopy of immunogold‐labelled erythrocytes
Authors: Heier H. E. , Namork E. , Kolberg J. , Falleth E.
Published: April 2008

SUMMARY. Binding patterns of mouse monoclonal antibodies (mAb) to P1, Pk, N, I, H, Y or A antigens were visualized in the backscatter electron imaging mode of a scanning electron microscope by indirect immunogold labelling. Experiments were performed at room temperature (RT) and at 4°C. In experiments with anti‐P1 and anti‐Pk, clusters of immunolabelling particles dominated the immunolabelling pattern much more at RT than at 4°C. By contrast, no clustering was seen with anti‐N, even at RT. Clustering was also observed at RT with anti‐I, anti‐H and anti‐Y, and on some Ax and A3 cells with anti‐A, but was much reduced at 4°C. Immunolabelling was stronger at 4°C than at RT with all mAb except anti‐N and anti‐A. The results indicate that glycolipid blood group antigens are more mobile in the membrane of intact erythrocytes at RT than at 4°C, and that the cells bind more antibodies to such antigens at 4°C than at RT. We suggest that antigen immobilization in the cold will reduce cross‐linking of antigens and hence increase the number of antibody molecules needed for epitope saturation, leading to increased binding of antibody in the cold. This may be the main reason for cold‐enhanced agglutination with human blood group antibodies.
The effect of glycosylation trimming enzyme inhibitors on monoclonal antibody recognition of α‐sialoglycoprotein epitopes
Authors: Strickland A. , Wilkins B. S. , Jones D. B.
Published: April 2008

SUMMARY. The human erythrocyte membrane contains four sialoglycoproteins, denoted α, β, γ and δ (also known as glycophorins A, C, D and B respectively), of which α‐sialoglycoprotein (α‐SGP) is the most predominant species. The extracellular portion of α‐SGP is heavily glycosylated with approximately 15 O‐linked carbohydrate side‐chains and a single N‐linked group.

We have used inhibitors of carbohydrate trimming enzymes to investigate the contribution of this single N‐glycan moiety towards the recognition of a range of antibody binding sites on α‐SGP. Two erythromyeloid cell lines, K562 and HEL, were cultured in the presence of these inhibitors and altered binding of antibodies to epitopes adjacent to the N‐glycan was observed. Digoxigenin‐coupled lectins were used to stain cytocentrifuge preparations and Western blots of cell lysates in order to confirm that modification of N‐linked carbohydrate side‐chains had been achieved. We suggest that the N‐glycan side chain of α‐SGP has a role in conferring conformational stability upon epitopes which lie in its vicinity.
Proteolytic activity during storage of platelets in plasma
Authors: Wallvik J. , Suontaka A. M. , Blombäck M.
Published: April 2008

SUMMARY. Proteolytic activity was studied in platelet concentrates (PC) stored in plasma at 22°C. In experiment 1, two PC with a higher (A) and a lower (B) white cell concentration were prepared from each of nine donors by centrifugation. Aliquots of the cell‐free plasma, PPP, were stored as a control. Samples for the assay of fibrinopeptide A (FPA), elastase, spontaneous proteolytic activity (SPA), kallikrein‐inhibiting activity, thrombin‐antithrombin complexes (TAT) and D‐dimers were collected initially and on days 1,3,5 and 7 of storage. Consumption of glucose, pH and concentrations of lactate dehydrogenase (LDH) and ATP were determined to investigate the metabolic status of the PC. The decrease in pH correlated to the leucocyte count, r= ‐0.74, P<0.001 and to the increase in LDH, r= ‐0.74, P<0.01. The levels of elastase and the SPA were consistently low in the PPP bags. In the PC elastase had increased by day 5 and the SPA by day 3; the levels in PC A were significantly higher than in PC B, P<0.01. The leucocyte count correlated with the elastase activity, r= 0.71, P<0.01, and with the SPA, r= 0.65, P<0.01. A minor increase in FPA was demonstrated while no TAT and D‐dimers could be detected. The cause of the formation of FPA was studied in experiment 2; three bags of PC and four of PPP were prepared from each of 16 donors. To the PC and three of the PPP bags either hirudin, aprotinin or no enzyme inhibitor (control) was added. To the fourth PPP bag extra CPD was added to evaluate the effect of pH. Hirudin decreased the formation of FPA (P<0.001) while aprotinin had no inhibitory effect; thus the formation was due in part, to thrombin activation. The levels of FPA were higher in the PPP controls, correlating with an increase in the pH.

We conclude that there is an activation of proteolytic enzymes in plasma during storage. The duration of storage, stability of pH and the concentration of white cells are important factors. The data give evidence for the benefit of a removal of leucocytes already in the preparation procedure.
Development, optimization and use of an enzyme linked immunosorbent assay (ELISA) to measure factor VIII antigen utilizing monoclonal antibodies
Authors: Hornsey V. S. , Drummond O. , Eaglesfield P. , Pepper D. S. , Prowse C. V.
Published: April 2008

SUMMARY. An enzyme linked immunosorbent assay (ELISA) has been developed to measure VIII:Ag in plasma and concentrates. The assay utilizes two commercially available monoclonal antibodies to VIII:Ag and provides an alternative to the established immunoradiometric assay (IRMA). It has the advantage of not requiring the use of radioactive material and human antibodies. The assay sensitivity is 0.006 u/ml and the interassay coefficient of variation is 6.3%. Forty‐eight samples with VIII:Ag levels ranging from 0.006 to 1.5 u/ml were assayed by both ELISA and IRMA. The coefficient of correlation between the two assays was 0.89. In addition to measuring human VIII:Ag, it is also possible to detect antigen in several animal plasma and sera.
Audit of the use of packed red blood cells in association with seven common surgical procedures
Authors: Pinkerton P. H. , Coovadia A. S. , Seigel C.
Published: April 2008

SUMMARY. The risks and costs associated with the transfusion of blood and its components have led to increasing demands for evidence of the appropriate use of blood components. We have examined the use of packed red blood cells (PRBC) in association with seven common, surgical procedures performed in 1987 and 1988 to establish patterns of use The information has formed the basis for surgical blood order schedules and autologous donation targets for these procedures. It has also been used to determine appropriate audit ‘triggers’ and the results of an audit using these ‘triggers’ are reported.
Molecular biology for platelet alloantigen typing
Authors: Williamson L. M. , Bruce D. , Lubenko A. , Chana H. J. , Ouwehand W. H.
Published: April 2008

SUMMARY. Hitherto, full investigation of patients with alloimmunization to platelet‐specific antigens has been difficult due to the limited availability of both typing reagents and panels of typed platelets. Following recent advances in the understanding of the molecular and genetic basis of platelet alloantigens, it is now possible to genotype individuals for the alleles coding for the epitopes of four platelet antigen systems (HPA‐1–4). This is based on the finding that the two alleles differ by oniy a single base pair substitution, resulting in one amino acid difference in the relevant platelet glycoprotein. The technique involves amplification of the relevant segments of genomic DNA from any nucleated cell by the polymerase chain reaction, followed by restriction fragment length polymorphism analysis. The technique allows investigation of thrombocytopenic individuals and fetuses/neonates, and can be readily applied to large‐scale typing of platelet donors.
Location of the granulocyte‐specific antigen LAN on the Fc‐receptor III
Authors: Metcalfe P. , Waters A. H.
Published: April 2008

SUMMARY. This report investigates the location of the binding site of a recently described granulocyte‐specific alloantibody, designated LAN, which causes alloimmune neonatal neutropenia. The LAN antigen is shown to be situated on the Fc‐receptor III (FcRIII).
U.K. multicentre study on blood donors for surrogate markers of non‐A non‐B hepatitis. Part I: Alanine transferase and anti‐HBc testing
Authors: Anderson N. A. B. , Raafat A. , Shwe K. H. , Barbara J. , Contreras M. , Fraser I. D. , Gunson H. H. , Martlew V. , Mijovic V. , Goldie D. J. , Seneviratne C. J. , Bendle A. , Briggs M. , Paver K.
Published: April 2008

SUMMARY. Blood samples from 9,215 blood donors in three U.K. centres (North London, Bristol and Manchester) were tested for their alanine aminotransferase (ALT) level and the presence of anti‐HBc and anti‐HCV. This paper presents the results of the ALT and anti‐HBc tests. The prevalence of ALT >45 IU/1 was 3.1% overall (North London 3.06%, Bristol 4.56% and Manchester 1.97%). Manchester results were skewed by the methodology used for ALT measurement, highlighting the need for standard test methods. Anti‐HBc was detected using the Wellcome enzymeimmunosorbent assay (EIA) and confirmatory testing was performed using a radioimmunoassay (RIA) and the Corecell haemagglutination assay. Repeat reactive rates were 0.9, 0.79 and 0.94% for North London, Bristol and Manchester, respectively, with an overall rate of 0.9%. The confirmed positive rate was 0.73, 0.53 and 0.65% for the three centres with an overall rate of 0.63%. Donors with an ALT >45 IU/1, or with confirmed anti‐HBc, were interviewed with a medical questionnaire for risk factors. The major contributing factors in donors with a raised ALT were alcohol consumption and obesity.
Detection of anti‐Fya, anti‐S and anti‐Jka in relation to the genotypes of the panel red cells: report of a U.K. NEQAS survey
Authors: Phillips P. K. , Whitton C. M.
Published: April 2008

Summary. Three antibody‐containing samples (anti‐Fya anti‐S and anti‐Jka), each at two dilutions, were distributed to U.K. hospitals and transfusion centres together with an antibody screening panel comprising red cells homozygous for the corresponding antigens. Participants in the study subjected the samples to their routine antibody screening procedures using both their own antibody screening panels and the screening panel provided.

A within‐group comparison of those participants using their own screening panels having a heterozygous expression of the antigens, with the same participants when using the screening panel provided, showed for five of the samples a greater detection rate in routine antibody screening procedures when using the panel provided, having homozygous expression of the corresponding antigens. The sixth sample, the most potent, was detected equally using both panels. The difference in overall detection rate is statistically significant (chi‐square test, 2p0.001).

The study shows that the use of red cells presumptively homozygous for Fya, S and Jka improved the detection of the corresponding antibodies.
ERIK, a low‐frequency red cell antigen of the MNS blood group system associated with Sta
Authors: Daniels G. L. , Green C. A. , Poole J. , Jerne D. , Smart E. , Wilcox D. , Young S.
Published: April 2008

Summary. A new low‐frequency red cell antigen, ERIK (MNS37), is associated with the Sta antigen of the MNS system. Four ERIK + propositi have been identified: all are St(a+). Thirteen other St(a+) samples, including one of the Mz type, and over 200 St(a‐) samples were ERIK‐. In two of the propositi ERIK is associated with an abnormal trypsin‐resistant M antigen; in the others it is associated with N, which is shown to be expressed weakly in one family. Immunoblotting with an antibody to an epitope common to glycophorin A (GPA) and glycophorin B (GPB) gave identical results with St(a+) EiRIK + and St(a+) ERIK‐cells, revealing GPA, GPB, an abnormal structure GPSta aggregates of these components. In the propositi with trypsin‐resistant M, GPStacarries the unusual M antigen, whereas in the M ‐ N + ERIK+ individual analysed by immunoblotting GPSta carries N. Immunoblotting with anti‐Sta anti‐ERIK showed that Sta is located on GPSta but that ERIK is located on GPA, presumably the GPA molecule encoded by the GYPA gene contiguous to the gene encoding GPSta.
Follow‐up study of anti‐hepatitis C virus antibodies in blood donors implicated in post‐transfusion non‐A, non‐B hepatitis
Authors: Mazda T. , Nakata K. , Bannai M. , Miyamura T. , Chiba J. , Ohba H. , Kaminuma Y. , Katayama T.
Published: April 2008

Summary. We retrospectively examined the antibodies to p22, a hepatitis C virus (HCV) nucleocapsid protein, and to c100‐3, a HCV nonstructural protein, in donors whose blood was transfused to patients who later developed post‐transfusion non‐A, non‐B hepatitis. Of 13 such blood donors, three seroconverted and three seroreverted with the anti‐c100‐3 test. In contrast, 12 of the 13 blood donors showed the same results at transfusion and follow‐up, and one donor showed seroconversion with the anti‐p22 assay. The follow‐up study shows that the anti‐p22 antibody test provides consistent results and is far more suitable for screening blood than the anti‐c100‐3 test.
Sensitivity of column agglutination technology in detecting unexpected red cell antibodies
Authors: Pinkerton P. H. , Chan R. , Ward J. , Coovadia A. S.
Published: April 2008

Summary. This study was conducted to compare a new microtube column agglutination technology (CAT) with a previously described tube saline‐indirect antiglobulin test (SIAT) with carefully defined performance in the detection of unexpected antibodies. On testing 117 sera from fresh and frozen stock containing antibodies detectable by SIAT, CAT failed to detect two examples of weak anti‐K. All other discrepancies between the two techniques involved antibodies generally regarded as clinically insignificant. Titration studies with anti‐D (concentration approximately 10 ng/ml) and 23 other antibodies, and studies with 10 weak antibodies of various blood group systems, showed the two techniques to be of similar sensitivity. Equivocal CAT results requiring repeated testing were found in 4·1% of specimens tested. We conclude that the CAT method adequately meets our requirements in sensitivity of detection of unexpected antibodies in pre‐transfusion testing and offers opportunities for savings in technical staff time.
Post‐transfusion purpura and delayed haemolytic transfusion reaction
Authors: Maślanka K. , Zupańska B.
Published: April 2008

Summary. Two women with post‐transfusion purpura (PTP) are presented, one with anti‐HPA1a and the other with anti‐HPA3a antibodies. Platelet‐specific antibodies were identified using the platelet immunofluorescence test (PIFT) and the monoclonal antibody immobilization of platelet antigens (MAIPA) assay. Lymphocytotoxic and red cell antibodies were also detected in both patients, the latter being responsible for a delayed haemolytic transfusion reaction (DHTR). In the patient with anti‐HPA1a antibody, red cell anti‐c alloantibody was found in the serum and in the eluate from red cells; it was active in the monocyte monolayer assay (MMA). The patient with anti‐HPA3a antibody had the red cell alloantibodies anti‐D, ‐M and ‐S detected in the serum, the last being responsible for DHTR.
Anti‐HAV prevalence in a U.K. urban blood donor population and the effect on human normal Ig provision
Authors: Howell D. R. , Thompson C. J. , Barbara J. A. J.
Published: April 2008

Summary. U.K. blood donors (873) were tested for anti‐HAV and 21% were found to be positive, with no obvious difference between the sexes. The positivity increased with age; 21% of donors between the ages of 30 and 50 years were positive compared with 3% of donors less than 25 years of age; in 1977, the rates were 47 and 10%, respectively. Older donors generally had lower titres of anti‐HAV which suggests that such donors were infected at an early age. Subsequent improvements in hygiene may have resulted in decreased anti‐HAV prevalence in young people. Titres of anti‐HAV were determined for the four donors who gave a history of jaundice and were seropositive: all four had high anti‐HAV levels. Plasma from donors with a history of jaundice may therefore provide a cost‐effective source for enhancing the potency of human normal Ig.
A blood grouping and antibody screening system using image analysis
Authors: Hewison C. , Hedley G. P. , Brittain M. , Doughty R. W.
Published: April 2008

Summary: An effective, modular automated blood grouping system has been developed which performs more effectively than the AG16C autogroupers and which uses readily obtainable reagents and disposables. Operating and maintenance costs are low by comparison with currently available equipment of similar capacity. The 99·2% success rate for blood grouping using this image analysis‐based system compares well with the 98·3% success rate on the AG16C autogroupers. Staffing requirements are reduced and the flexible modular approach allows this Newcastle system to be used for a range of serological functions, including grouping, antibody screening and screening for antigen donors. The ability to operate with small sample numbers without long start‐up times and the relatively low capital cost makes the Newcastle system an option that could be considered by the larger hospitals as well as transfusion centres. The sensitivity of the system could be developed to include compatibility testing as a feature of the system.
Immunochemical characterization of the anti‐B response in an ABO‐incompatible blood transfusion: presence of antibodies recognizing glucosylceramide
Authors: Rydberg L.
Published: April 2008

Summary. A male patient of blood group A was accidentally given two units of group B blood when operated upon because of injury. This resulted in an immediate haemolytic transfusion reaction. The patient was treated with exchange transfusions in order to remove the incompatible red cells and thus prevent renal damage. Two weeks after the transfusion, an anti‐B humoral immune response appeared. The antibodies were mainly IgG and IgA types, of IgG3 and IgA1 subclasses. The anti‐B antibodies seemed specific for the B trisaccharide [Galα1–3(Fucα1–2)Gal] as tested by radioimmunoassay and chromatogram binding assay. No antibodies recognizing part of the core saccharide chain could be detected, i.e. antibodies capable of differentiating between B type 1 and B type 2 chains did not occur. An unsuspected finding was that the patient had IgG antibodies recognizing glucosylceramide (and weakly also galactosylceramide) before and after the transfusion. These antibodies were still present 9 years after the ABO‐incompatible blood transfusion.
The production and evaluation of two human monoclonal anti‐D antibodies
Authors: McCann‐Carter M. C. , Bruce M. , Shaw E. M. , Thorpe S. J. , Sweeney G. M. , Armstrong S. S. , James K.
Published: April 2008

Summary. The passive administration of anti‐D immunoglobulin is effective in preventing Rh alloimmunization and therefore preventing haemolytic disease of the newborn. However, diminishing availability and increased demand render an alternative source of anti‐D desirable. We report the production of two human monoclonal IgGl antibodies specific for Rhesus antigen D. Data are presented on the serological reactivity of these antibodies and on their ability to promote in vitro biological activity. Antibody derived from both lymphoblastoid cell lines and the hetero‐hybrid cell lines were compared for both ESD‐1 and ESD‐4. The ability of the antibodies to detect rhesus related antigens in animal tissue was also assessed. We present some evidence for altered reactivity of the ESD‐4 antibody following fusion of the lymphoblastoid parent line to a murine plasmacytoma.
Haemolytic anaemia caused by anti‐Pra following rubella infection
Authors: Kadota Y. , Fujinami S. , Tagawa Y. , Sato M. , Miyazaki H. , Shiozaki Y. , Inoue K. , Ishida T. , Miyamoto A. , Okubo S. , Yasunaga K.
Published: April 2008

Summary. A 24‐year‐old male was admitted to Kansai Medical University Hospital, complaining of fever, skin exanthema, jaundice, brown urine and lymphadenopathy.

The patient was diagnosed as having haemolytic anaemia caused by a cold agglutinin following rubella infection.

The cold agglutinin of the patient reacted strongly with group OI red blood cells (RBC), Oi cord RBC, Oi adult RBC and neuraminidase‐treated RBC, and much weaker with protease (papain, ficin, bromelin)‐treated RBC; it was identified as anti‐Pra.

Cold agglutinins of anti‐Pra specificity following rubella infection in adults have rarely been reported.
Half‐strength citrate CPD and new additive solutions for improved blood preservation. 2. The effect of storage at ambient temperature before component preparation and different means of supplying glucose to the red cells
Authors: Högman C. F. , Eriksson L. , Gong J. , Vikholm K. , Debrauwere J. , Payrat J. M.
Published: April 2008

Summary. In current anticoagulant‐additive systems used for the preparation of blood components an early loss of erythrocyte 2,3‐bisphosphoglycerate (BPG) during storage is unavoidable. This increases reversibly the oxygen affinity of the haemoglobin of red blood cells (RBC). Using half‐strength citrate CPD solution (0·5CPD) as anticoagulant and a red‐cell additive solution (RAS 2) containing citrate, adenine, phosphate, mannitol and glucose, it was possible effectively to counteract this undesirable effect without loss of adenine nucleotides. The initial mean BPG level was maintained for 4 weeks and the total adenine nucleotide concentration for 7 weeks. The initial BPG level decreased upon prolonged hold of the whole blood at ambient temperature to 85–90% of normal after 4 h, 70% after 12 h and 50% after 22 h. Both preparation and storage haemolysis were lower in 0·5CPD blood processed into RAS‐2‐suspended RBC than in CPD blood processed into SAGM‐suspended cells.

The possibility of adding glucose to the red cells via the primary anticoagulant instead of the additive solution was tested. The glucose concentration had to be increased to three times that in normal CPD to supply the red cells with sufficient glucose for 6–7 weeks of storage. However, this caused increased haemolysis, particularly after prolonged hold of the whole blood at ambient temperature before component preparation. The results therefore indicate that the glucose supply should be in the additive solution. Less citric acid in the 0·5CPD anticoagulant, an increase in intracellular pH on addition of the new additive solution and supply of phosphate seem to be major factors in explaining the effects. The new combination of anticoagulant and red‐cell additive solution is a clear improvement for blood component preparation and storage.
Print‐on‐demand blood‐group labels
Authors: Houston S. R. , Muircroft J. H. , Barr A. , Mitchell R.
Published: April 2008

Summary. The requirement within the U.K. and Europe to use a standardized black‐and‐white label format from 1 April 1992 in conjunction with an increasing awareness of quality and information technology (IT) pointed us towards the use of print‐on‐demand label systems. The introduction of this system has enabled a greater level of control over the issue, usage and quality of the labels attached to blood and blood products. The system is versatile, secure and responsive to future alterations in blood‐group labelling protocols.
Discharge haematocrit as clinical indicator for blood transfusion audit in surgery patients
Authors: Goodnough L. T. , Vizmeg K. , Riddell J. , Soegiarso R. Wida
Published: April 2008

SUMMARY. Three cohorts of elective surgical patients were reviewed in order to develop a method in which the discharge haematocrit can serve as a clinical indicator for a subsequent study of the use of blood transfusion therapy. Three different levels of discharge haematocrit were evaluated: 36, 33, and 30% for ‘generous’, ‘intermediate’, and ‘strict’ criteria, respectively. Discharge haematocrits (%, mean ± SD) for patients not transfused were 29.6±4.6, 33.7 ± 5.0, and 33.6 ± 3.4 for three different surgical groups (cardiac, orthopaedic, and urological surgical patients). When discharge haematocrits greater than 33% (‘intermediate’) were considered excessive due to previous transfusion, the prevalence of patients identified was 9, 6.5 and 13%, respectively. We found no relationship between the length of stay in hospital and the number of blood units transfused or patient discharge haematocrit levels. When the length of stay of patients identified by exceeding the clinical indicator was compared to that of patients not identified, orthopaedic and urological surgical patients showed no difference; however, cardiac surgical patients who exceeded the clinical indicator had shorter hospital stays compared to patients who were not so identified. We conclude the following:

1 The discharge haematocrit can be used as a clinical indicator for a subsequent review of use in order to evaluate the appropriateness of blood transfusion therapy.

2 The prevalence of patients identified who exceeded the clinical indicator, among three elective surgical patient groups, suggests that this indicator is applicable across elective surgical categories in order to target transfusion medicine education programmes and clinical outcome studies.

3 Additional factors important to the ‘transfusion trigger’, such as blood lost during hospitalization, may need to be included with the discharge haematocrit as clinical indicators in order to evaluate blood transfusion therapy in this setting.
Informed consent for blood transfusion as a transfusion medicine educational intervention
Authors: Goodnough L. T. , Hull A. L. , Kleinhenz M. E.
Published: April 2008

SUMMARY. The aim here was to determine the effectiveness of a transfusion medicine educational intervention in a medicine core clerkship program.

Third‐year medical students enrolled in their medicine core clerkship rotations at tertiary care hospitals affiliated with our institution underwent a two‐part educational intervention that incorporated a transfusion medicine curriculum within the context of the medicolegal, ethical and educational elements of informed consent. Part one was a 1‐h didactic session on standards of practice for red blood cell transfusion. Part two was a 90‐min multidisciplinary workshop on informed consent. The effectiveness of the educational intervention was analysed by an objective structured clinical evaluation.

The student group receiving the educational intervention scored significantly higher than in the comparison group (65.8 ± 9.2 vs. 54.1 ± 10.56, P <0.001). When student scores were used to determine changes in student response patterns over time, the largest change occurred in identifying possible other options to allogeneic blood transfusion.

These results suggest that a transfusion medicine curriculum using an informed consent model can be used effectively as an educational intervention in a medicine core clerkship programme.
A study of anti‐hepatitis C positive blood donors: the first year of screening
Authors: MacLennan S. , Moore M. C. , Hewitt P. E. , Nicholas S. , Barbara J. A. J.
Published: April 2008

SUMMARY. In the U.K., blood donations have been routinely screened for anti‐HCV since September 1991. In order to get the most epidemiological benefit from these extensive screening data, the histories obtained at counselling from donors confirmed to be anti‐HCV positive, ‘indeterminate’ and falsely positive have been analysed in detail. In addition, the associations with potential risk factors have been investigated by comparing these groups of donors with a control group of 771 routine donors bled on one day during the study, at North London Blood Transfusion Centre. This paper documents the prevalence and demography of HCV infection in asymptomatic blood donors, to assess varicus possible sources of infection and the association between liver function test results and alcohol consumption in donors. One in 1400 previously untested donors was confirmed positive for anti‐HCV. Age (the group 30–49 years being highest), tattooing and intravenous drug use in both sexes, ear‐piercing in males and blood transfusion in females were all significantly associated with an increased risk of HCV infection. Intravenous drug use proved to be the factor most strongly associated with risk. Liver function tests (alanine aminotransferase) were elevated in a significant number of donors confirmed to be anti‐HCV positive but no clear correlation between alanine aminotransferase level and either time since infection or alcohol consumption was found. Alcohol consumption was significantly higher in donors confirmed to be anti‐HCV positive and was particularly marked in those admitting to previous intravenous drug use. Although donors confirmed to be anti‐HCV positive had a 5–10 times greater chance of non‐Caucasian ethnic origin compared with controls, the association with ethnic origin was not as marked as it was for HBsAg positive donors.
Sensitivity of the platelet immunofluorescence test (PIFT) and the MAIPA assay for the detection of platelet‐reactive alloantibodies: a report on two U.K. National Platelet Workshop Exercises1
Authors: Allen D. L. , Chapman J. , Phillips P. K. , Ouwehand W. H.
Published: April 2008

SUMMARY. This report presents the results of two National Workshop exercises which were designed to evaluate interlaboratory and interassay variation in sensitivity of techniques used for the detection of platelet‐reactive alloantibodies. Most workshop participants used the platelet immunofluorescence test. Sensitivity of this assay was improved when fluorescence was measured by flow cytometer rather than by microscope. The MAIPA assay was found to be highly sensitive but required considerable technical expertise, and the choice of antiglycoprotein II/IIIa (CD61/41) monoclonal antibody was found to have a significant effect on its ability to detect anti‐HPA‐la.
Risk of hepatitis C in patients who received blood from donors subsequently shown to be carriers of hepatitis C virus
Authors: Ayob Y. , Davidson J. I. , Baxter A. , Jordan A. , Yap P. L. , Gillon J.
Published: April 2008

SUMMARY. A retrospective study was undertaken to identify recipients of blood from donors subsequently shown to be positive for hepatitis C virus using second‐generation tests and polymerase chain reaction. The main aims were to determine the numbers of such recipients who were still alive and traceable, and to determine the risk of infection in this group. The feasibility and workload of this procedure, which is currently not practised in the U.K. or U.S.A., was also assessed.

In the first six months of routine testing 42,697 donors were tested. Of 20 confirmed to be HCV‐positive, 15 were regular donors. Eighty‐three components were prepared from 63 anti‐HCV positive previous donations from these donors. In all, nine recipients were found to be alive. All were positive for anti‐HCV. We conclude that although this retrospective procedure is time‐consuming and difficult, substantial numbers of infected recipients can be identified. The availability of treatment for chronic hepatitis C for such patients should encourage transfusion services to reassess current policies on the hepatitis C retrospective.
HCV antibodies in Hungarian blood donations. Experiences collected by ELISA tests, immunoblot assays and polymerase chain reaction and protocols for donor management
Authors: Héjjas M. , Medgyesi G. A. , Falus A. , Hajnal A. , Forster T. , Szabó J.
Published: April 2008

SUMMARY. Routine screening of Hungarian blood donors for anti‐HCV commenced in the second half of 1992. Before this, five available anti‐HCV ELISA kits were compared in pilot studies. In the first series, 831 random donor samples were tested by one of the tests and the 12 (1.4%) reactives found were retested by the other four. Six of the reactives were positive in all ELISA. In the second series, 325 samples from donors with elevated transaminase levels were tested by all five kits. Forty‐four were found to be reactive by one or more of the tests and 32 (10%) were positive in all five assays. Samples concordantly reactive in the ELISA were positive in second or third generation recombinant immunoblot assay (RIBA 2 or RIBA 3); those that gave discordant results were indeterminate or negative. Eleven concordantly reactive samples from the second series were HCV RNA positive by polymerase chain reaction (PCR). In the first period of screening with Abbott ELISA 2 a repeat‐reactivity rate of 0.98% was observed in 171,106 samples tested. Reactives were retested for supplementary testing by Wellcozyme anti‐HCV. Donors reactive in both tests and strongly reactive (ELISA ratio (ER) = optical density/cut off 2.5) in either of them were permanently deferred. Those negative in the supplementary ELISA or weakly reactive (1.0<2.5) in both tests were subjected to RIBA 2. On the basis of RIBA, positive donors were permanently deferred, indeterminates were excluded for 1 year and negatives were readmitted. In 1992, 1,347 supplementary tests were completed; 824 (61%) of the respective donors were permanently deferred, 218 (16%) were deferred for 1 year and 305 (23%) were readmitted.
Three examples of Rh haemolytic disease of the newborn with a negative direct antiglobulin test
Authors: Heddle N. M. , Wentworth P. , Anderson D. R. , Emmerson D. , Kelton J. G. , Blajchman M. A.
Published: April 2008

SUMMARY. Typically the serological diagnosis of alloimmune haemolytic disease of the newborn (HDN) includes a positive direct antiglobulin test on the infant's red cells, and the presence of an IgG red cell alloantibody in both maternal and cord sera. HDN with a negative direct antiglobulin test has been reported with anti‐A and anti‐B, but not with other red‐cell alloantibodies.

In this report we describe four examples of HDN in infants whose red cells had a negative direct antiglobulin test. The first case was diagnosed retrospectively when the infant was admitted to hospital aged 3 weeks with severe anaemia and cardiac failure, and subsequently died. Maternal and infant sera were both shown to contain anti‐C: however, the direct antiglobulin test on the infant's red cells was negative. Approximately 1 year later the mother of this infant gave birth to triplets: soon after birth one of the triplets required an exchange transfusion, one had hyperbili‐rubinaemia, and the third was unaffected. Anti‐C and anti‐e were detectable in the maternal serum at this time. The most probable Rh genotypes of the two affected infants were R1R2 (CDe/cDE), while the Rh genotype of the unaffected infant was R2R2 (cDE/cDE). Anti‐c was implicated as causing HDN in a fourth infant (from a different family) who was a hydropic stillborn. The direct antiglobulin test on fetal blood was negative and other causes of non‐immune hydrops were excluded. These four infants provide evidence that the direct antiglobulin test may be negative in some severely affected and even fatal cases of HDN.
Post‐transfusion purpura due to HPA‐1a immunization in a male patient: response to subsequent multiple HPA‐1a‐incompatible red‐cell transfusions
Authors: Gabriel A. , Laßnigg A. , Kurz M. , Panzer S.
Published: April 2008

SUMMARY. We report a male patient who developed post‐transfusion purpura. Thrombocytopenia did not recur when intravenous IgG was given before the transfusion of platelet‐depleted washed red‐cell concentrates.
Monocyte monolayer assay (MMA) reactivity of alloantibodies reacting by the manual polybrene technique but not by an antiglobulin test
Authors: Lown J. , Willis J.
Published: April 2008

SUMMARY. The manual polybrene technique (MP) has been shown to be useful in pretransfusion testing as a rapid method capable of detecting a wide range of red cell antibodies. The clinical significance of alloantibodies reacting by the MP but not by an antiglobulin test (MP +/AGT‐) has not been determined.

The monocyte monolayer assay (MMA) is an established in vitro technique for assessing the likely clinical significance of red cell antibodies. This was used to examine 21 MP +/AGT‐ alloantibodies and 41 alloantibodies reacting by a LISS antiglobulin method (LISS‐AGT).

Two of the 21 MP +/AGT‐ antibodies (9.5%) produced positive results by the MMA compared with 26/41 (63.4%) of LISS‐AGT‐reactive antibodies. The incidence of positive reactivity of the latter antibodies in the MMA was similar between those that were reactive and nonreactive by the MP.

These results suggest that antibodies reacting by MP and not by an antiglobulin test are less likely to be clinically significant.
A pharmacokinetic study of an ion‐exchange solvent—detergent‐treated high‐purity factor VIII concentrate
Authors: Ludlam C. A. , Lowe G. D. O. , Mayne E. E.
Published: April 2008

SUMMARY. An ion‐exchange chromatography purified factor VIII concentrate (Liberate) that had undergone a solvent—detergent viral inactivation treatment was compared with an intermediate‐purity concentrate (Z8), which was terminally heated at 80°C for 72 h, in 15 haemophilia A patients in a blinded crossover pharmacokinetic study. Both products achieved a peak level close to that predicted (100IU/dl for Liberate and 103IU/dl for Z8) and there were no significant differences in the recoveries achieved nor of any of the other pharmacokinetic parameters. We conclude that the pharmacokinetic properties of factor VIII: C, following solvent—detergent treatment and ion‐exchange chromatography, are equivalent to those of the lower purity terminally heat‐treated product (Z8), and it is therefore likely to be a clinically efficacious concentrate.
Anti‐D quantification by flow cytometry: an alternative to the Auto Analyser?
Authors: Austin E. B. , McIntosh Y. , Hodson C. , Lee D.
Published: April 2008

SUMMARY. A flow cytometry method for quantifying levels of anti‐D is described. The method is based on the indirect antiglobulin test and uses an FITC Fab anti‐human IgG reagent for detection of bound anti‐D. A standard curve is generated from the fluorescence measured by flow cytometry using the British Standard anti‐D preparation 73/515. The fluorescence obtained from test samples by flow cytometry is converted into IU/mL anti‐D, using the standard curve. In a limited study with 42 serum samples the assay showed good correlation with results obtained using the AutoAnalyser for sera with anti‐D levels less than 100 IU/mL. The method is simple to perform and has potential as a replacement for the AutoAnalyser.
Analysis of donor‐deferral pattern in a voluntary blood donor population
Authors: Chaudhary R. K. , Gupta D. , Gupta R. K.
Published: April 2008

SUMMARY. A reterospective analysis of records of the deferred donors from 1 October 1992 to 31 December 1993 was performed. Of 14,269 prospective blood donors (13,030 males and 1,239 females), 2,431 (16.4%) donors were disqualified for various reasons: 8‐1% of the donors were deferred for non‐pathological causes while 91‐9% were deferred for medical reasons. The most common cause for non‐pathological deferral was volunteers attending below the minimum acceptable age (5‐2%). Abnormal findings on physical examination accounted for 57‐2% of the deferrals in which low body weight was the most common finding (32‐3%) followed by low Hb (18‐6%). A past history of jaundice was the leading cause for deferral on medical interview.

Numerous prospective donors are currently being deferred based on empirically derived criteria. By developing strategies to identify and rationalize donor selection criteria, the blood transfusion services should be able to decrease unnecessary deferrals.
A multicentre assessment of the specificity of ten anti‐HBc screening tests
Authors: Hughes W. , Barr A. , Dow B. C. , Follett E. A. C. , Barbara J. A. J.
Published: April 2008

SUMMARY. Samples from 1828 donations were screened for anti‐HBc at seven sites in the UK using kits supplied by 10 manufacturers. Only 10 (0.55%) donations were considered to have true anti‐HBc reactivity and these were detected by all 10 kits. Additional markers of HBV infection were found in nine of these 10 donations. Additional reactives were found by all kits, the number ranging from 1 to 43.

In the four more specific kits, the 10 true reactives were clearly distinguished from the ‘false reactives’ by the strength of the reaction. It is concluded that the reliance on a single ELISA test for anti‐HBc diagnosis is unwise. The use of a second test known to be more specific than the screening ELISA is recommended.
Blood product costing: relationship to price and clinical efficacy
Authors: Trenchard P. M. , Dixon R.
Published: April 2008

SUMMARY. Detailed information is provided about primary product costing and price issues as they affect transfusion manufacturing practice and clinical transfusion practice. Product price is shown to have a crucial influence upon clinical practice and associated research. By focusing particularly upon cost‐benefit analysis of blood product transfusion therapy a substantive conclusion is drawn that price should equal the associated manufacturing cost. Clinical outcome studies relate clinical efficacy to the manufacturing specification of the product, which should therefore determine the product cost. Thus, the true manufacturing cost is the sum of all the process activity costs that create the final product specification, e.g. red cell number + volume reduction + leucocyte reduction + microbiological safety, for processed red cells. Sometimes different product specifications may compete for a single activity cost, e.g. one‐spin processing achieves volume reduction and leucocyte reduction for processed red cells but also plasma removal for protein fractionation. A method for understanding the relative clinical importance of different products is described, which guides the cost allocation process. Furthermore, for some products there is uncertainty about the clinical benefits of some components of the specification, e.g. leucocyte load and immunomodulation, and a method is described for ranking this quality‐uncertainty level objectively. The optimal costing model must ensure that the product with the highest uncertainty ranking is assured a high degree of cost stability. These concepts prepare the way for a Quality Associated Costing model for blood products that correlates with clinical efficacy.