bluuebunny/crossref_metadata_2025 huggingface dataset:
Quality of platelet concentrates irradiated with UVB light: effect of UV dose and dose rate on glycocalicin release and correlation with other markers of the platelet storage lesion
Authors: Bessos H. , Murphy W. G. , Robertson A. , Vickers M. , Seghatchian M. J. , Tandy N. P. , Cutts M. , Pamphilon D. H.
Published: April 2008

Summary. The amount of membrane‐associated glycoprotein Ib in platelet concentrates (PCs) irradiated with a high dose of UVB light has been shown to be significantly reduced after 48 h storage. We recently corroborated this finding when we noted an increase in the supernatant levels of glycocalicin (GC, a major segment of glycoprotein Ib) in UVB‐treated PCs during storage. The aim of the present study was to determine whether GC release was related to both the UV dose and the rate of dose delivery. Plateletpheresis concentrates obtained from five donors were pooled and split into five equal parts. Four of these were treated with 7500 and 15000 mJ/cm2 UVB using two prototype UV sources with differing rates of dose delivery; namely, Baxter (BAT) and British Aerospace (BAC) cabinets, with the latter having the slower rate of delivery. On days 1 and 5 of storage, GC levels in the supernatants of PCs were determined by ELISA. Moreover, the following parameters were also assessed: platelet and WBC count; hypotonic shock response (HSR) and platelet aggregation response to ADP, ADP +collagen, ADP + arachidonic acid and ristocetin; pH; supernatant levels of lactate, glucose, von Willebrand factor (vWf) and β‐thrornboglobulin (βTG). The results revealed an association of GC release with UVB dose using both UV sources, although this was more apparent in the BAC system, in which glycocalicin release at day 5 of storage was as follows (μg/ml, mean ± SD): 4·8±0·3 and 9·5±3·6 at 7500 and 15000 mJ/cm2 respectively. Moreover, at 15000 mJ/cm2, PCs treated in the BAC system exhibited significantly higher levels of GC than those treated in the BAT system: 9·5±3·6 and 4·8± 3·6 respectively at day 5 of storage (P= 0·05). This differential GC increase in the BAC was coupled with a decrease in HSR and a significant increase in lactate and βTG levels compared with the BAT system. In contrast to the GC results, vWf supernatant levels in PCs treated with UVB were decreased relative to non‐treated PCs of the same origin. Moreover, GC release correlated significantly with various standard tests of platelet function indicating its importance as a quality indicator for the investigation of the platelet storage lesion. Our results show that UVB not only increases GC release in a dose/rate‐dependent manner but that it may also affect the quality of irradiated PCs and their shelf life.
Monoclonal antibodies against Kell glycoprotein: serology, immunochemistry and quantification of antigen sites
Authors: Parsons S. F. , Gardner B. , Anstee D. J.
Published: April 2008

Summary. Monoclonal antibodies BRIC 18, BRIC 68, BRIC 107 and BRIC 203 recognize high‐frequency epitopes absent from erythrocytes expressing the Kophenotype. BRIC 107 has anti‐k (K2)‐like specificity. BRIC 203 has a unique specificity denoted anti‐Kpbc. All four monoclonal antibodies identify an Mr, 95 600 erythrocyte membrane protein by immunoprecipitation from radio‐iodinated erythrocytes. In quantitative binding studies using IgG it is estimated that there are from 2000 (BRIC 18) to 4000 (BRIC 68) copies of the Kell glycoprotein per erythrocyte. Using Fab fragments the estimates are in the range 4000 (BRIC 18) to 18 000 (BRIC 68) copies. In competitive binding assays the four epitopes defined by the BRIC monoclonal antibodies fall into two non‐overlapping groups. The first group comprises BRIC 18, BRIC 68, BRIC 203 and an antibody (6–22) with anti‐K14 specificity. The second group contains BRIC 107 and two further anti‐k‐like monoclonal antibodies (BS45 and OSK5). The results suggest that the polymorphisms encoded at the K/k and Kpa/Kpb/Kpc loci may be located in two spatially distinct regions of the Kell glycoprotein(s).
Improved sensitivity in the monocyte‐mediated ADCC assay with red cell targets
Authors: Kirkwood J. , Trueman K. , Fletcher A.
Published: April 2008

Summary. Monocyte‐mediated antibody‐dependent cellular cytotoxicity (ADCC) assays are being increasingly employed in blood transfusion to predict the severity of haemolytic disease of the new‐born (HDN). In our hands the monocyte‐mediated ADCC assay resulted in very high spontaneous release of 51Cr, making the assay unreliable. The addition of bovine serum albumin (BSA) to the buffer used to wash red blood cells (RBC) was found to reduce the spontaneous release to acceptable levels, improving the sensitivity of the assay. The modified assay was used to test 15 monoclonal anti‐D antibodies showing that the assay results are reproducible.
Clinical efficacy of a highly purified SD‐treated factor IX concentrate prepared by conventional chromatography
Authors: Goudemand J. , Marey A. , Caron C. , Wibaut B. , Mizon P.
Published: April 2008

Summary. A highly purified (HP) FIX concentrate was used in 12 patients with haemophilia B in order to prevent bleeding following surgery. The HP FIX concentrate (CRTS, Lille, France) was prepared by ion‐exchange chromatograpy and was SD treated. The patients underwent orthopaedic surgery (3), neurosurgery (3) or multiple dental extraction (6). They were treated for 2–18 days and received 8500 to 102000 FIX units. The mean FIX:C recovery was 0·92 ± 0·20%/U/kg. Satisfactory haemostasis was provided in all patients. None of them experienced thrombotic manifestations. There was no change in FDP or soluble fibrin complexes. Prothrombin fragments one plus two (F1 + 2), thrombin‐antithrombin III complexes (TAT) and cross‐linked fibrin degradation products (D‐dimer) remained in the normal range in one patient treated for 2·5 days for dental surgery. In contrast, elevated levels of F1+2, TAT and D‐dimer were observed in one patient undergoing a bilateral hip arthroplasty treated for 16 consecutive days with the same lot of HP FIX concentrate as the previous patient. There was no seroconversion for HIV, HBV and HCV. These results indicate that the HP FIX concentrate provides satisfactory haemostasis in surgery while avoiding excessive thrombogenic risks. However, some clinical circumstances, such as orthopaedic surgery, are known to be associated with especially high thrombotic risks that must be taken into account in case of FIX replacement.
Serum ferritin, blood donation, iron stores and haemochromatosis
Authors: Worwood M. , Darke C.
Published: April 2008

Summary. Serum iron and ferritin concentrations were measured in 1,532 regular blood donors from South Wales who were undergoing HLA typing prior to registration on the British Bone Marrow and Platelet Donor Panel. Serum transferrin concentrations were determined for donors with serum iron concentrations > 24 µmol/1. There were 25 donors with transferrin saturations > 50% and 11 with transferrin saturations > 60%. There were five donors with serum ferritin concentrations > 200 µg/1 (women) or > 300 µg/1 (men). Two of the male donors had transferrin saturations > 50% and serum ferritin >300 µg/1 on repeat blood samples and are being treated by venesection. Donors with HLA‐A3 did not differ from those without A3 in serum iron or ferritin concentrations. Even in the group of donors who were apparently homozygous for A3 there were neither abnormal serum iron nor ferritin concentrations.

Although it is well established that measurements of transferrin saturation are required to detect homozygous haemochromatosis (HFE) in its earlier stages, the number of ‘false‐positive’ results is likely to be unacceptably high for screening blood donors. Serum ferritin assays should identify donors with HFE and iron overload before the onset of liver damage. With two million regular donors and 300,000 new donors each year, a significant proportion of the U.K. population will be screened within 10 years. The assay of serum ferritin identifies donors with low levels of storage iron who are at risk of developing iron‐deficiency anaemia. Furthermore, donation frequency may be increased for those donors with higher ferritin concentrations when blood supplies are low.
STEM, a new low‐frequency Rh antigen associated with the e‐variant phenotypes hrS––– (Rh: ––– 18, ––– 19) and hrB––– (Rh: ––– 31, ––– 34)
Authors: Marais I. , Moores P. , Smart E. , Martell R.
Published: April 2008

Summary: The new low‐frequency antigen STEM was identified when a Cape coloured woman with an unknown antibody in her plasma (which agglutinated the red cells of her husband and a minority of other individuals) gave birth to a baby suffering mildly from haemolytic disease of the new‐born. Most, but not all, examples of anti‐STEM distinguish different strengths of STEM antigen on the red cells of different people; the different strengths are inherited. Family studies established that STEM was inherited as a Mendelian dominant character. STEM subdivides hrs––– (Rh: ––– 18, ––– 19) and hrB––– (Rh: ––– 31, ––– 34) red cells into two types: STEM + and STEM –––. The manually calculated lod score for STEM being associated with the Rh system is 3·91 and LIPED calculated lod score 4·35. The International Society of Blood Transfusion has allocated STEM the Rh number 49.
Half‐strength citrate CPD and new additive solutions for improved blood preservation. I. Studies of six experimental solutions
Authors: Högman C. F. , Eriksson L. , Gong J. , Payrat J.‐M. , Debrauwere J.
Published: April 2008

Summary: Poor stability of plasma factor VIII in whole blood and loss of erythrocyte 2,3‐bisphosphoglycerate (BPG) during red cell storage are limitations with systems for blood component preparation in current use. This study presents attempts to improve post‐collection storage conditions in both these respects using half‐strength citrate CPD solution (0·5CPD) for blood collection, which has been shown by others to improve the stability of factor VIII, and some compositions of hypotonic additive solutions for red cell storage containing citrate, adenine, mannitol, and phosphate. Guanosine was also included in some of the media. The erythrocyte BPG concentration was maintained at a normal level for 3–4 weeks with the best of the tested compositions. Total adenine nucleotide concentration was maintained at the original level for 49 days and adenosine triphosphate for 28 days. Spontaneous storage haemolysis was low, 0·31% (mean) ± 0·08‐0·10% (SD) after 49 days in the two best compositions. The intracellular pH was 0·2‐0·3 pH units higher than the extracellular pH at the beginning of storage, but this difference gradually diminished and disappeared after 4–5 weeks. We suggest two likely explanations of the effects: the maintenance of intracellular pH at a level sufficiently high not to impair BPG synthesis until after several weeks of storage, and a sufficient supply of phosphate needed in the synthesis of organic phosphate compounds. The content of citrate was selected such that the total amount supplied to a patient in a massive transfusion, when using a combination of 0·5CPD plasma and red cell suspension, would be smaller than that provided by a transfusion of CPD whole blood.
Further complexities of the Rh antigen D disclosed by testing category DII cells with monoclonal anti‐D
Authors: Lomas C. , McColl K. , Tippett P.
Published: April 2008

Summary: The reaction pattern of monoclonal anti‐D with category DII cells differed from those of other category D cells. DII cells express epDl, epD2, epD3, epD5, epD6/7 and epD8 but lack epD4 and a new epitope epD9. The new epitope, epD9, is proposed to explain the failure of some monoclonal anti‐D (previously considered to be anti‐epD3) to react with DII cells.
Characterization of blood‐group‐active erythrocyte membrane glycoproteins with human antiseras*
Authors: Spring F. A.
Published: April 2008

Summary: Antisera to high‐ and low‐incidence blood‐group antigens were used in immunoblotting and immune precipitation studies to identify novel erythrocyte membrane components and to assign antigens to known proteins. Several antibodies identified well‐characterized membrane proteins which are widely expressed on other cells and tissues. The Cromer system antigens were found to reside on the complement regulatory protein decay accelerating factor (DAF). Antigens of the Indian collection were located to the cell adhesion molecule, CD44. The Cartwright system antigens were assigned to acetylcholinesterase, the function of which, on erythrocytes, remains unclear. Two novel blood‐group‐active glycoproteins were identified. One carries the Scianna system antigens whilst the second carries the high‐incidence antigens, Gya, Hy and Joa. The low‐incidence antigens, Dha and Rd, were assigned to Glycophorin C and to the Scianna‐active glycoprotein, respectively. The existence of Cromer‐null, DAF‐deficient erythrocytes greatly facilitated the study of the function of DAF on erythrocytes. Location of the YT locus and hence of the AChE gene to chromosome 7q22 may be of significance in leukaemias and myelodysplasias since this region is a mutational ‘hot spot’ in these disorders. The novel proteins identified, for which no monoclonal antibodies are available, may also prove to be of functional significance on erythrocytes or on other cells and tissues. Several of the rare phenotype cells, such as the In(a ––– b –––) cells, Gy(a–––) cells and the Sc‐null cells, may prove to be of great value in defining the function of these molecules on erythrocytes, in the way that Inab cells have been for studying the function of DAF.
Auto Analyzer quantification, monocyte‐mediated cytotoxicity and chemiluminescence assays for predicting the severity of haemolytic disease of the newborn
Authors: Hadley A. G. , Garner S. F. , Taverner J. M.
Published: April 2008

Summary. The value of quantitative and functional assays in predicting the severity of haemolytic disease of the newborn (HDN) was assessed. Sera containing anti‐D were obtained from 27 pregnant women who subsequently delivered D‐positive babies. Antibody levels were quantified by Auto Analyzer (AA) and functional activity was determined by an antibody‐dependent cell‐mediated cytotoxicity (ADCC) and chemiluminescence test (CLT). The severity of haemolytic disease was graded according to cord Hb levels and transfusion requirements of the babies. In eight cases unaffected by HDN, six results by AA (over 5IU/ml), one by CLT and two by ADCC falsely predicted the occurrence of HDN. In nine cases of mild to moderate HDN requiring transfusion therapy, there was one false negative result by AA, three by CLT and two by ADCC. All assays correctly predicted severe HDN in the remaining 10 cases. The ADCC assay and CLT showed good correlation (t= 0·69, P < 0·001) and the tests were of comparable sensitivity. The data suggest that while all assays are capable of discriminating severe HDN, AA results may falsely predict HDN in unaffected babies. Although the CLT may not discriminate some mild cases of HDN, ADCC and CL tests are less susceptible to false positive results in unaffected cases.
Hepatitis B core antibody screening of voluntary blood donors: an extended pilot study using a modified passive haemagglutination assay
Authors: Martlew V. J. , Rogan P. D. , Shepherd A. J. N. , Firth S. A.
Published: April 2008

Summary. Screening for hepatitis B surface antigen (HBsAg), even by the most sensitive techniques, fails to detect all carriers of the hepatitis B virus (HBV). The presence of hepatitis B core antibody (anti‐HBc) in the absence of HBsAg is a common finding in donors implicated in cases of post‐transfusion hepatitis B (PTHB), and viral DNA may also be demonstrated in many of these individuals. An extended pilot study of routine screening of all donations for anti‐HBc in addition to HBsAg was introduced into the Mersey and North Wales Regional Transfusion Service in November 1991 to improve surveillance for carriers of HBV. In order to reduce costs a modified passive haemagglutination assay was evaluated and found to have a sensitivity of 99% and specificity of 98% compared with a conventional assay. In the first 6 months 60 530 donations were tested and 12 (0–02%) were found to have anti‐HBc in the absence of HBsAg or adequate antibodies to HBsAg (anti‐HBs). These sera will later be submitted for polymerase chain reaction (PCR) testing in order to determine their infectivity or otherwise by the detection HBV DNA sequences.
Post‐transfusion purpura: a survey of 12 Danish cases with special reference to immunoglobulin G subclasses of the platelet antibodies
Authors: Taaning E. , Svejgaard A.
Published: April 2008

SUMMARY.The clinical and immunological data of 12 Danish cases of post‐transfusion purpura (PTP) are summarized. All patients but one were women. All except one had thrombocytopenic purpura which occurred 4–11 days after transfusion, usually with a nadir of the platelet count below 10times1091‐1. The typical haemorrhagic symptoms were cutaneous bleedings, melena and haematuria lasting from 3 to 12 days. The time until normalization of platelet count varied from 3 to 50 days after onset. One patient had recurrence of PTP and one patient died due to intracranial haemorrhage.

Ten of the HPA‐la negative patients (83%) had platelet‐specific HPA‐la antibodies and two HPA‐lapositive individuals had anti‐HPA‐lbantibodies. In 10 patients, HLA antibodies were also detectable and in one patient a delayed haemolytic transfusion reaction due to anti‐E was seen.

All of seven patients investigated had anti‐HPA antibodies of both IgGl and IgG3 subclasses during the thrombocytopenic period while all of 10 patients had anti‐HPA of only the IgGl subclass after recovery from PTP Thus, the destruction of autologous platelets in PTP seems to be associated with the presence of anti‐HPA of the IgG3 subclass which may be of importance in the pathogenesis of PTP.
Human platelet alloantigen typing: PCR analysis is not a substitute for serological methods
Authors: Morel‐Kopp M.‐C. , Clemenceau S. , Aurousseau M.‐H. , Schlegel N. , Kaplan C.
Published: April 2008

SUMMARY. Currently, five platelet alloantigen (alloAg) systems have been established (HPA‐1, ‐2, ‐3, ‐4, ‐5). Three of these are expressed on the glycoprotein (GP) IIb‐IIIa complex, HPA‐1, HPA‐3 and HPA‐4, inherited in an autosomal codominant mode. Recent investigations of the molecular basis of these platelet alloantigen systems have shown that only one nucleic acid base substitution in the genes encoding for GP IIb and GP IIIa is responsible for the polymorphism. This substitution is reflected in a difference in restriction enzyme recognition allowing platelet alloantigen typing by restriction fragment length polymorphism (RFLP) analysis of DNA amplified by the polymerase chain reaction (PCR). To validate the PCR technology for platelet typing, we have compared PCR‐RFLP with monoclonal‐antibody‐specific immobilization of platelet antigens (MAIPA). For this purpose, we have studied different Glanzmann thrombasthenic families and particularly heterozygous individuals, who are not lacking GP IIb‐IIIa, as a model to detect the occurence of discrepancies between these two technologies. In two families, we have found differences between molecular biology and serological methods with the lack of expression of one antigen on the platelet membrane surface. In the first family, the abnormality is related to the HPA‐1 alloantigen system with three informative members; in the second, the HPA‐3 alloantigen system is concerned with two informative members. Considering these results, there may not always be a perfect correlation between molecular biology and serological methods, as an unknown molecular defect could interfere with the PCR results and lead to false platelet typing. In cases of suspected post‐transfusion purpura (PTP) or fetuses at risk for neonatal alloimmune thrombocytopenia (NAITP), great care must be taken in the interpretation of the results if only PCR‐RFLP techniques are used.
Human platelet antigen‐1 (Zw) typing of fetuses by analysis of polymerase chain reaction‐amplified genomic DNA from amniocytes
Authors: Simsek S. , Christiaens G. C. L. M. , Kanhai H. H. H. , Beekhuis J. R. , Bleeker P. M. M. , Vlekke A. B. J. , Goldschmeding R. , Kr. von dem Borne A. E. G.
Published: April 2008

SUMMARY. Prenatal typing for the human platelet antigens‐1 (HPA) permits identification of a fetus at risk for neonatal alloimmune thrombocytopenia (NAITP) in cases of HPA‐1 incompatibility in which the father is heterozygous for the HPA‐la antigen. Diagnostic cordocentesis and phenotyping of the fetal platelets are used for this purpose. We applied allele‐specific restriction enzyme analysis on polymerase chain reaction (PCR)‐amplified DNA purified from amniocytes. This assay allows early second trimester typing for HPA‐1 alleles. We were able to determine the genotype of three fetuses at risk. Iatrogenic fetal loss is lower with amniocentesis than with cordocentesis. Therefore, this technique is a welcome addition to the antenatal management of NAITP.
A comparative study of plateletpheresis using Baxter Autopheresis C and Haemonetics PCS Plus
Authors: Walton J. D. , Caffrey E. A. , Allain J. P.
Published: April 2008

SUMMARY. This study compared plateletpheresis on the Haemonetics PCS Plus (PCS Plus) and the Baxter Autopheresis C (Auto C) using the same 100 selected donors. The number of packs meeting UK BTS/NIBSC specification (>2.2 times 1011 platelets per pack) was achieved by 99% of PCS Plus and 82% of Auto C procedures. The positive correlation found between donor precount and final platelet yield was better for the PCS Plus. Both machines met U.K. specification for white‐cell contamination but this was significantly greater for the Auto C. Plasma yields were similar.

As a result of this study we chose to use the PCS Plus for routine plateletpheresis in our unit. This has enabled us not only to comply with UK BTS/NIBSC specifications for apheresis platelets easily and cost effectively but also to meet our own higher specification (2.75 times 1011 platelets per pack) using existing staff and without extending the working day.
Prevalence and epidemiological characteristics of hepatitis C in Scottish blood donors
Authors: Crawford R. J. , Gillon J. , Yap P. L. , Brookes E. , McOmish F. , Simmonds P. , Dow B. C. , Follett E. A. C.
Published: April 2008

SUMMARY. All blood donors in Scotland who were found to be infected with hepatitis C virus (HCV) in the first 6 months of routine testing of all donations for anti‐HCV were contacted. Those who attended were counselled, a history of exposure to risk was sought, and blood was taken for alanine aminotransferase (ALT) level as a measure of liver function. The epidemiological features were then correlated with the virological findings and ALT.

In the period under study between September 1991 and February 1992, 180658 blood donors attended. The prevalence of HCV infection was 0.088%. Of the 151 donors who attended for counselling, 101 (68%) were male. Intravenous drug use was the most common risk activity (39%), followed by previous blood transfusion (15.2%), other parenteral exposure (11.2%) and heterosexual contact with a parenterally infected partner (8.6%); 29.1% of donors gave no history of possible exposure.

Elevated ALT levels were found in 59%. ALT levels were higher in donors with HCV types 1 and 3 than in HCV type 2 or non‐viraemic donors.

The prevalence of HCV in Scottish blood donors is thus relatively low. This may relate to the effectiveness of donor selection procedures, but donors with risk activities which should debar them continue to donate. The combination of ALT and PCR appears to be useful in counselling and assessing infected donors.
Infectious disease markers in autologous and directed donations
Authors: Pink J. , Thomson A. , Wylie B.
Published: April 2008

SUMMARY. Autologous collections are strongly advocated by the New South Wales Red Cross Blood Transfusion Service (BTS) and have increased more than sevenfold since 1988. Directed donations, although not promoted, have also increased during this time. The prevalence of infectious disease markers (HIV, hepatitis C, hepatitis B and syphilis) in donations collected by the BTS from different donor groups including overall volunteer homologous, first‐time volunteer homologous, autologous and directed were evaluated over a 42‐month period. Donations from first‐time volunteer homologous donors had the highest prevalence of hepatitis B and C. Autologous donations had a significantly higher prevalence of hepatitis B, hepatitis C and syphilis compared with overall volunteer homologous donations. The percentage of directed donations testing positive for either hepatitis B or C was higher than overall volunteer homologous donations, but not statistically significant. This study demonstrates that donations from first‐time donors are the least safe, that the crossover of autologous blood into the volunteer homologous pool decreases the safety of that pool and suggests that directed donations may not be as safe as volunteer homologous donations and cannot be generally advocated at this time.
Blood group antigens defined by the amino acid sequences of red cell surface proteins
Authors: Anstee D. J.
Published: April 2008

SUMMARY. The antigens of 18 blood group systems are expressed on proteins that are intrinsic to the red cell. The proteins which carry the antigens of these systems have been identified and primary sequence information is available for all but two (SC, DO). Several different functional groups are evident. Antigens of the DI, CO, RH, XK and JK systems are located on proteins which have the structure of membrane transport proteins. The FY antigens mark a cytokine receptor. The IN, LW, XG antigens are associated with molecules which have adhesion functions and the LU glycoprotein also has a structure which suggests a role in adhesion. YT and KEL antigens are located on cell surface enzymes and the CR and KN antigens on molecules involved in complement regulation. Finally, the MN and GE antigens are located on sialic acid‐rich glycoproteins (glycophorins A, B and C/D respectively), a group of molecules which do not, as yet, have a clearly defined function. The molecular basis of antigens in several blood group systems have been defined and shown to depend upon the amino acid sequence.
Multicentre trial of antepartum low‐dose anti‐D immunoglobulin
Authors: Lee D. , Rawlinson V. I.
Published: April 2008

SUMMARY. Routine antenatal Rh immunopro‐phylaxis would substantially increase the use of anti‐D Ig in the U.K. As availability of anti‐D Ig is one factor influencing a decision to introduce routine antenatal prophylaxis, a trial was undertaken to test the efficacy of a lower dose of anti‐D Ig than that used in earlier studies. RhD‐negative primigravidae were randomized as controls or recipients of two doses of 250 iu of anti‐D Ig given at 28 and 34 weeks gestation. Blood samples were tested at delivery and at 6 months postpartum for the presence of immune anti‐D, and again later if results were equivocal. Nine (1.5%) out of 595 control patients had immune anti‐D at follow‐up at 6 months and later; 4 (0.78%) of 513 treated women were immunized. It was concluded that, while two doses of 250 iu of anti‐D Ig may reduce allo‐immunization, they are not as effective as two doses of 500 iu in a previous trial.
A case of unexplained mild Rh (D) haemolytic disease in utero
Authors: Bromilow I. M. , Downing I. , Walkinshaw S. A. , Welch C. R. , Duguid J. K. M.
Published: April 2008

SUMMARY. This report describes the case of a patient with a history of HDN complicated by fetal losses, in which the alloantibody in this particular pregnancy did not appear to cause HDN in utero. No protective HLA‐DR antibodies could be demonstrated, and transport of IgG across the placenta appeared to be normal. The infant's red cells possessed a normal D antigen and his mononuclear phagocyte system appeared unimpaired. However, the number of molecules of IgG bound in vivo per fetal red cell was below the level usually associated with significant haemolysis and HDN.
Quantification of anti‐c in haemolytic disease of the newborn
Authors: Kozlowski C. L. , Lee D. , Shwe K. H. , Love E. M.
Published: April 2008

SUMMARY. Anti‐c is an important Rh antibody that causes haemolytic disease of the newborn (HDN). We have carried out a retrospective analysis of the clinical outcome of pregnancy in 120 mothers with anti‐c. Of these, 100 gave birth to c‐positive infants, of whom 14 had severe HDN requiring exchange transfusion. In all of these, the maternal level was 9.5iu/ml or greater. Of the 29 women with anti‐c levels of 9.5iu/ml or above, in addition to the 14 with severely affected infants, 15 had infants requiring only phototherapy or no treatment. Our observations suggest that when the anti‐c level is below 7.5iu/ml, the fetus is unlikely to be seriously affected and invasive obstetric intervention is unnecessary.

Of the 120 women studied, 50% had had a blood transfusion, in most cases for obstetric complications in a previous pregnancy. Although it was not possible to attribute alloimmunization to blood transfusion rather than previous pregnancy in any individual case, this observation points to the value of routine c typing as part of antenatal screening, so that c‐negative blood can be selected.
Use of standardized protease enzymes for antibody screening of blood donor samples with the microplate system AutoAnalyzer
Authors: Mazda T. , Makino K. , Yabe R. , Nakata K. , Fujisawa K. , Ohshima H.
Published: April 2008

SUMMARY. Papain, bromelin and ficin can be standardized by the casein method for use in red blood cell antibody screening tests. The minimal and optimal enzyme activity for detecting blood group antibodies in donors, using the new Automated Pre‐Transfusion Blood Testing System Olympus PK7200 by the two‐stage method for all three enzymes, is one casein unit. One casein unit of proteinase activity changed the red blood cell surface charge to a low plateau value as measured by electrophoretic mobility and sialic acid content, and removed sterically inhibiting structures of surface protein as detected by SDS‐PAGE.
Transfusion practice in urological surgery: modifications in association with the introduction of an autologous blood donor programme
Authors: Pinkerton P. H. , Klotz L. H.
Published: April 2008

SUMMARY. The transfusion records of 122 urological patients who had deposited one or more units of autologous blood were analysed. The 95 patients undergoing radical prostatectomy were arbitrarily divided into two cohorts (the first 47 patients enrolled, and the last 48 enrolled) to assess changes in use of blood. The avoidance of allogeneic transfusion in radical prostatectomy was 64% in the first cohort studied, and increased to 81% in the second. There was a significant decline in the mean absolute amounts of blood used from 4.47 to 2.54 units per patient from the first to second cohorts. Introduction of an autologous blood donor programme has been associated with a reduction in total blood use for radical prostatectomy, and a decrease in the number of patients exposed to allogeneic blood. The validity of an autologous blood‐order schedule of 3 units for radical prostatectomy is supported.
Isolation of IgA1 from human serum by affinity chromatography using an immobilized extract of the albumin gland of Helix pomatia
Authors: Booth J. R. , Munks R. , Sokol R. J.
Published: April 2008

SUMMARY. An extract of the albumin gland of Helix pomatia was linked to Sepharose‐4B and used to prepare IgA from group O human serum; immunoelectrophoresis showed that the preparation was free of IgG and IgM. From studies with specific IgA subclass antisera and by comparison with the activity of jacalin‐produced material the Helix pomatia extract was found to be IgA1 specific. The preparation had red cell anti‐A,B specificity and was suitable for standardizing and controlling anti‐human IgA reagents. Preparations using six different carbohydrates as eluants inhibited the agglutination reaction between anti‐human IgA and IgA‐coated red cells to varying degrees. The pattern of reactions suggested that N‐acetyl glucosamine was the IgA binding site for Helix pomatia; this differed from its blood group A determinant (N‐acetyl galactosamine) which was the same as that for the IgA1 reactive component of jacalin.
Use of autologous blood in support of orthopaedic surgery using a hospital‐based autologous donor programme
Authors: Pinkerton P. H.
Published: April 2008

SUMMARY. The use of autologous blood in support of orthopaedic surgery has been assessed for 296 patients; 150 underwent hip arthroplasty, 69 underwent knee arthroplasty, 37 underwent spinal procedures and 40 underwent miscellaneous operations. Overall, 87% of patients received no allogeneic blood and 23% of the autologous blood collected was not used. For hip and knee arthroplasty, there appears to be an increased willingness to transfuse patients when autologous blood is available, and a decreased proportion of patients receiving more than 3 units for hip arthroplasty and 2 units for knee surgery, when compared with an audit of blood use

when almost all blood used was allogeneic. Wastage of autologous blood in support of spinal surgery was 66%, prompting a review of ordering practices. Assessment of avoidance of allogeneic transfusion by the standard schedule of 3 units for hip arthroplasty and 2 units for knee arthroplasty appears justified by the calculation that collection of an additional unit in each case would avoid allogeneic transfusion in 11 (5%) more patients with the unnecessary collection of 208 units.
Blood transfusion in beta thalassaemia major
Authors: Rebulla P.
Published: April 2008

SUMMARY. Conventional treatment of beta thalassaemia major is based on regular blood transfusion from early childhood. Maximum effectiveness of transfusion therapy depends on the following. (1) Availability of safe blood. Donation programmes should aim at retaining repeat donors, who carry decreased risk of transmitting blood‐borne infections. Donors should be screened with laboratory tests performed to the highest possible standard of quality. Selection of safe donors can be improved by the adoption of questionnaires containing direct questions on risk factors for transfusion transmissible infections. (2) Use of good quality red blood cells, which should be leucodepleted, preferably by filtration, that can be carried out at the bedside. (3) Regular evaluation of blood transfusion indices, including mean level of haemoglobin maintained, annual blood requirement, daily haemoglobin fall, mean transfusion interval, transfusion reaction rate. This can be assisted by the use of a computerized patient record. (4) Maintenance of a permanent record of the patient's blood group genotype (including at least Rh, Kell, Kidd and Duffy systems) and any red cell antibodies that develop. This is mandatory to ensure optimal survival of transfused red cells. (5) Continuous monitoring of transfusion transmissible infections. (6) Vaccination against hepatitis B of all suitable patients. (7) Intensive iron chelation. This should be done by regular subcutaneous administration of desferoxamine B. Oral chelators, which are currently under laboratory and clinical evaluation, are not yet available for general use.
AIDS awareness in blood donors in North India
Authors: Choudhury N. , Singh P. , Chandra H.
Published: April 2008

SUMMARY. Prevention of AIDS can only be achieved by a successful public awareness programme. This study was carried out to establish the level of awareness of AIDS and HIV infection through blood transfusion among Indian blood donors of various socioeconomic groups. A questionnaire consisting of 20 questions pertaining to various aspects of AIDS and HIV infection was circulated to 1012 voluntary blood donors. The responses were categorized as good, average and poor knowledge according to the number of correct responses. The majority of the donors were males (93.5%), married (69.7%) and belonged to the Hindu community. Of the donors, 44.4% were educated, 49.9% were highly educated and 5.8% were illiterate. Overall, only 205 (20.3%) showed good awareness of AIDS and the majority of these 168 (16.6%) were highly educated. Approximately 80% of our population did not have sufficient knowledge about AIDS or the danger of contracting as well as disseminating this disease in the community.

One hundred and ninetyone (18.9%) donors were in the higher income group and only 13 of them showed good awareness. Most of the donors, 384 (38.0%), were government white collar workers and belonged to the middle income group. In this group 154 had good awareness.

The majority of donors with good awareness (176 out of 205) preferred to receive HIVtested banked blood or blood from their own relatives during emergencies. None of the blood donors had any knowledge of autologous blood donations.

This study showed that awareness of AIDS was not satisfactory in Indian society. More intensive public awareness campaigns by the Government with the help of Non‐Governmental Organizations is required in India to prevent an explosive AIDS situation in the near future.
Relationship of serum alanine aminotransferase (ALT) to body mass index (BMI) in blood donors: the need to correct ALT for BMI in blood donor screening
Authors: Ramesh V. , Saraswat S. , Choudhury N. , Gupta R. K.
Published: April 2008

SUMMARY. A study was carried out on 1,028 voluntary blood donors to see how body mass index (BMI) correlated with the serum alanine amino transferase (ALT) activity. The mean ALT (U/1) values were 19.35, 27.63, 40.79 and 54.41 in the four BMI categories of ≤20, 20.1–25, 25.1–30 and >30, respectively. This study showed that the mean serum ALT level of obese subjects (BMI > 30kg/m2), compared with the two categories of normal subjects (i.e. BMI ≤ 20 and BMI = 20.1–25 kg/m2), was increased by 2.8 and 1.96 times, respectively. Compared with the BMI group ≤ 20, there was a gradual per cent increase in the mean serum ALT in the three different BMI groups: 20.1–25 (+133%), 25.1–30 (+196%) and >30 kg/m2 (+280%). This indicates the need to correct ALT values for BMI for blood donor screening, instead of using actual ALT values.
Screening for IgA deficiency on the Olympus PK7100 by haemagglutination inhibition
Authors: Mohabir L. A. , Rees T. J.
Published: April 2008

SUMMARY.Severe or life‐threatening anaphylactic reactions during blood transfusion to patients with anti‐IgA, though infrequent, can be avoided by transfusing blood products prepared from IgA deficient donors. This report describes the development and use a passive haemagglutination assay adapted for use on the Olympus PK7100 blood group analyser. Donor plasma samples identified as being presumptive IgA deficient were further screened by a manual passive haemagglutination inhibition technique and finally confirmed by means of a quantitative enzymelinked immunosorbent assay. Of 5723 donors tested simultaneously for IgA deficiency, blood grouping and red cell antibody screening, six (1 in 954) were confirmed as IgA deficient (i.e. < 100ng/ml of IgA). The test is simple and cost effective and can be used to establish a database of IgA‐deficient donors.
Human platelet antigen‐2 and ‐3 genotyping by PCR‐SSP
Authors: Metcalfe P. , Bleeker P. M. M. , von dem Borne A. E. G. Kr. , Ouwehand W. H.
Published: April 2008

SUMMARY. Allele‐specific PCR using sequence specific primers (PCR‐SSP) is a simple and reliable technique to detect point mutations in genes. We have developed a PCR‐SSP to enable the detection of a C‐T mutation at position 482 of the GPIb gene and a T‐G mutation at position 13962 in exon 26 of the GPIIb gene. These point mutations are at the basis of the HPA alloantigens 2a, 2b and 3a, 3b respectively. One primer of each primer set has a 3′ nucleotide complementary to the DNA sequence coding for one allele. PCR product is only produced when the corresponding DNA is present and thus the genotype is determined by the presence or absence of a band in agarose electrophoresis of PCR products. A second set of primers in the same reaction yields a product regardless of the HPA genotype to control the efficiency of the PCR amplification. The HPA‐2 and ‐3 genotypes determined in this way were in strict concordance with those established by conventional genotyping using PCR followed by restriction enzyme digestion (PCR‐ASRA). PCR‐SSP is a rapid and reliable technique that can be used for the determination of alleles which code for platelet alloantigens.
In‐line filtration of platelet concentrates obtained with the OmnixTM blood cell separator
Authors: Moog R. , Müller N. , Nieper A.
Published: April 2008

SUMMARY. The quality of platelet concentrates (PC) obtained with the blood cell separator OmnixTM was investigated before and after in‐line filtration. PC were filtered 2h (protocol A) and 4h (protocol B) after the termination of apheresis. Platelet (PLT) yield after filtration was similar in both protocols (median 3.7 vs. 3.4 times 1011). Median white blood cell (WBC) contamination after leucocyte depletion was 0.07 times 106 (range 0.02–3.27 times 106) in protocol A and 0.06 times 106 (range 0.02–2.1 times 106) in protocol B. Glucose, lactate, lactate dehydrogenase, morphology score and pH value were not statistically different before and after filtration in both protocols. We conclude that in‐line filtration results in sufficient leucocyte depletion of the PC. The prefiltration storage time did not influence the studied parameters of product quality.
Monoclonal anti‐D specificity and Rh D structure: criteria for selection of monoclonal anti‐D reagents for routine typing of patients and donors
Authors: Jones J. , Scott M. L. , Voak D.
Published: April 2008

SUMMARY. The Rh blood group system is the next most important to the ABO system in terms of its clinical significance in blood transfusion. It is vital to the safe, efficient practice of transfusion medicine that Rh D phenotyping tests are selected, executed and interpreted correctly. However, the Rh D blood group antigen has been shown to be subject to many phenotypic variations, and different reagents and typing techniques vary in their ability to detect these variants. The range of D‐positive phenotypes are reviewed in terms of their reactivity with monoclonal antibody reagents and their clinical significance. In view of the available evidence, it is suggested that patient typing can be safely achieved by the duplicate use of one high‐avidity or two very similar IgM monoclonal anti‐D reagents that detect most variants except category DVI in simple tube or microplate saline tests. Antiglobulin testing for weak D should not be carried out on patient samples. Donor typing can be safely achieved by the use of the same monoclonal, used in parallel with a polyclonal anti‐D reagent that detects DVI on sensitive automated equipment.
Platelet alloantigens HPA‐1, ‐2, ‐3, ‐5 and ‐6b in Finns
Authors: Kekomäki S. , Partanen J. , Kekomäki R.
Published: April 2008

SUMMARY. The human platelet alloantigens HPA‐1, ‐2, ‐3, ‐5 and ‐6b in the Finnish population were determined using allele specific restriction analysis (PCR‐ASRA) for HPA‐1, ‐2, ‐3 and ‐5 and monoclonal antibody immobilized platelet antigen (MAIPA) assay for HPA‐1, ‐3a, ‐5b and ‐6b. No discrepancies were observed between the results obtained with the PCR‐method and those obtained serologically. The gene frequencies obtained from 200 unrelated Finns were 0.86 and 0.14 for HPA‐1a and ‐1b, 0.91 and 0.09 for HPA‐2a and ‐2b, 0.59 and 0.41 for HPA.3a and ‐3b and 0.95 and 0.05 for HPA‐5a and ‐5b. The frequency of the HPA‐5b allele (10%) is lower in Finns than in Central‐ or South‐European populations (20–30%). The HPA‐1, ‐2 and ‐3 frequencies did not deviate from those observed in other European populations. The rare HPA‐6b antigen was observed in three of 127 individuals from south‐eastern Finland (2.4%), which suggests that the frequency of this allele in Finland is higher than previously thought.
Red cell antibodies in pregnancy: there is no ‘critical titre’
Authors: van Dijk B. A. , Dooren M. C. , Overbeeke M. A. M.
Published: April 2008

SUMMARY. The purpose of this study was to determine the predictive value and reliability of using a ‘critical titre’ when assessing the ability of red cell alloantibodies to cause haemolytic disease of the newborn. Titration studies and clinical follow‐up of 418 antenatal cases where the mothers had red cell antibodies were studied retrospectively. The antibody specificities were anti‐D (n= 359), anti‐c (n= 34), anti‐E (n= 19) and anti‐K (n= 6). Depending on the titre being lower or higher than 16 in the indirect antiglobulin test, the severity of disease was established on the given therapy.

Anti‐D antibodies with a titre 16 were present in 20% of all cases associated with transfusion need of the child; for anti‐c, ‐E and ‐K the figure was 4%. Titres ≥ 16 resulted in both groups in 50% of the cases in phototherapy only, or no therapy at all. Titres are therefore not reliable indicators for predicting the severity of haemolytic disease of the newborn. Neither should they be used as a guide to whether or not antenatal intervention is indicated. Alternative quantitative or functional assays that measure cytotoxic lysis or phagocytosis or a combination of both should be performed instead.
Frequency and characteristics of post‐transfusion hepatitis in Greece with emphasis on hepatitis C: comparing second‐ and third‐generation assays
Authors: Makris K. , Kouvelis V. , Drakopoulos I. , Oikonomou E. , Maniatis A.
Published: April 2008

SUMMARY. In order to evaluate the role of hepatitis C virus (HCV) in post‐transfusion hepatitis (PTH) in Greece we prospectively followed 143 transfusion recipients, receiving 790 units of blood and/or products from 789 donors, between October 1989 and December 1991. The mean number of units transfused per patient was 5–52. PTH was observed in 18 patients (12–59%). One patient (0–70%) developed hepatitis B, in four (2–80%) hepatitis could be attributed to CMV infection, 10 (6–99%) developed hepatitis C and three (2–10%) showed only raised alanine aminotransferase (ALT) levels. The risk of PTH per 1000 units transfused was 22–8. The patient who developed hepatitis B (PTH‐B) was transfused with four units, one of which was positive for anti‐HBc and anti‐HBe. Seven of the 10 patients (70%) who developed hepatitis C (PTH‐C) were transfused with at least one unit seropositive in the anti‐HCV screening with 2nd‐generation tests (ELISA‐2 and RIBA‐2), whereas 9/10 of PTH‐C cases (90%) were transfused with at least one unit positive in 3rd‐generation assays. Of the three patients who showed only ALT elevation, none was transfused with anti‐HCV seropositive blood, although one of them was transfused with at least one unit with elevated ALT levels. We conclude that: (1) the incidence of PTH in Greece remains high, (2)screening of all donations for anti‐HCV with an ELISA‐2 does not exclude transmission of HCV and (3) ELISA‐3 and RIBA‐3 seem to be more sensitive in blood donor screening and in detecting seroconversions than ELISA‐2 and RIBA‐2.
A matter for debate: the risk of bovine spongiform encephalopathy to humans posed by blood transfusion in the UK
Authors: Dealler S.
Published: April 2008

Summary. If human infection with bovine spongiform encephalopathy (BSE) were to occur, donated peripheral blood from humans that might have become infected from eating adequate quantities of food containing BSE should, until evidence is available to the contrary, be assumed to contain the human form of the disease. The chance of disease transfer to a blood recipient in 1995, which might in turn cause clinical disease with an incubation period of 20 years, is calculated. Transfusion is calculated to be a potential cause of a maximum of only 0.2% of clinical cases of Creutzfeldt‐Jakob disease (CJD) in the UK population if the BSE epidemic were to spread to humans. Prospective epidemiological techniques would be unlikely to demonstrate any such minor contribution that blood transfusion might make to CJD incidence.