bluuebunny/crossref_metadata_2025 huggingface dataset:
Platelet Storage Lesions in Second‐Generation Containers: Correlation with in vivo Behavior with Storage up to 14 Days
Authors: Holme S. , Heaton W.A.L. , Whitley P.
Published: March 2009

Abstract. The relationship between in vivo behavior and in vitro characteristics of 59 platelet concentrates (PC) stored for up to 14 days in a synthetic medium or in CPDA‐1 plasma was systematically investigated. 25 paired studies (1 study was incomplete) were performed comparing platelets suspended either in the synthetic medium or CPDA‐1 plasma with 5 days (n = 5); 7 days (n = 10); 10 days (n = 5); and 14 days (n = 5) of storage. In addition, 10 control studies were performed with freshly prepared PC (6–24 h) in CPDA‐1 plasma. Both percent recovery and survival estimations showed decreases with increasing storage duration, irrespective of storage medium used. In both media, with prolonged storage, the platelet survival curves not only became shorter, but also increasingly exponential, suggesting that in vitro storage caused progressive damage to the platelets present in circulation. Survival curves of platelets suspended in synthetic medium remained more linear, indicative of less random damage during storage. Mean population lifespan (MPL) of the stored PC was determined by the area below the survival curve divided by the mean percent recovery for the fresh PC, which was 55%. MPL decreased from 4.5 days (fresh PC) to 0.4 days after 14 days of storage in plasma, with a 50% reduction (t½) estimated at 7.2 days of storage. MPL t½ for PC stored in the synthetic medium was estimated to be 8.8 days. The decrease in in vivo viability with prolonged storage was paralleled with loss of energy‐dependent in vitro parameters such as hypotonic shock response, shape change with ADP, and ATP levels, and with increased lactate levels. Although the length of the storage period was shown to be the major factor responsible for the variability in MPL observed in this study, multiple linear‐regression analysis showed also that platelet discoid shape, as measured by the shape change, and lactate production were independent in vitro parameters with significant predictability of the in vivo viability. Combined with days of storage in the regression equation, these parameters were found to ‘explain’ 78% of the variability of the MPL found in this study.
Effectiveness of the Imugard E Leukocyte Removal Filter for Preparation of Leukocyte‐Poor Concentrated Red Cells
Authors: Takebe Michikazu , Tsubokura Michio , Amaki Ichita
Published: March 2009

Abstract. Concentrated red cells (CRC) were filtered through a new leukocyte removal filter, the Imugard E, which consists of a polyvinyl alcohol porous sheet. CRC were filtered through the Imugard E with neither priming before filtration nor rinsing after filtration. Leukocyte removal was 99.1 ± 0.6, 99.6 ± 0.3 and 99.6 ± 0.4% on the 1st, 5th and 10th day after blood collection, respectively. Platelet removal was 96 ± 2, 81 ± 6 and 85 ± 3% on the 1st, 5th and 10th day, respectively. Red cell recovery was 86 ± 2,86 ± 1 and 86 ± 1% on the 1st, 5th and 10th day, respectively. Filtration time was 3.9 ± 0.8, 5.8 ± 0.9 and 6.1 ± 0.8 min on the 1st, 5th and 10th day, respectively. Direct filtration of CRC through the Imugard E resulted in no significant changes in the ATP or 2,3‐DPG concentrations, and no hemolysis due to filtration was noticed. It may be concluded that the Imugard E is a good filter that is simple to use and effective in leukocyte removal.
Erythroid HLA Class I Expression in 300 Patients with Haematological, Renal and Rheumatological Disorders
Authors: Ranasinghe W.A.E.P. , Giles C.M. , Pusey C.D. , Hows J. , Ritter M.A. , Walport M.J.
Published: March 2009

Abstract. The expression of HLA class I was assessed on erythrocytes by haemagglutination with monoclonal antibodies to monomorphic epitopes on the heavy and light (β2‐microglobulin) chains. Previously, enhancement of HLA class I expression was observed on erythrocytes of many patients with systemic lupus erythematosus (SLE) and chronic lymphatic leukaemia (CLL), and we have now tested erythrocytes from patients (and 130 normal controls) with other auto‐immune diseases and renal and haematological disorders. The striking enhancement in patients with SLE and CLL was confirmed. A significant increase in expression was also observed in aplastic anaemia patients following bone marrow transplantation and in renal patients with primary glomerulonephritis who had received a transplant. No class I was expressed by erythrocytes from many patients with inherited haemoglobinopathies and high reticulocyte counts, which suggests that the enhancement in SLE patients cannot be accounted for by immature or young erythrocyte populations. The distribution of HLA‐A and ‐B types in the patients with enhanced class I expression did not relate to those antigens previously detected more frequently on erythrocytes, B7(Bga), B17(Bgb), A28(Bgc), B8 or A10, and the enhancement was not associated with any particular HLA types.
A Simple Plasma Anticoagulant‐Exchange Method to Increase the Recovery of Factor VIII in Therapeutic Concentrates
Authors: Cumming A. M. , Wensley R. T. , Winkelman L. , Lane R. S.
Published: March 2009

Abstract. Donor blood, primarily anticoagulated by acid citrate dextrose formula A (ACD‐A), was separated by means of the HemaScience Autopheresis CTM plasmapheresis device. The citrated plasma was collected directly into a solution of heparin and calcium chloride to achieve a final plasma‐ionised calcium concentration of approximately 2 mM, and a heparin concentration of 1.0 IU/ml. Heparin at this concentration provided adequate anticoagulation, and did not result in insoluble cryoprecipitates. Three pairs of donor‐matched 4‐kg plasma pools (anticoagulant‐exchanged variant and ACD‐A‐anticoagulated control) were constructed and subsequently fractionated to an intermediate stage. The mean recovery of factor VIII from 3 anticoagulant‐exchanged pools (394 IU/kg) was 23% greater than the mean recovery from the matched control pools (319 IU/kg). This increased recovery was not achieved at the expense of specific activity.
Moderate and Severe Reactions during Autologous Blood Donations Are No More Frequent than during Homologous Blood Donations1
Authors: McVay P. A. , Andrews A. , Hoag M. S. , Polan D. , Skettino S. , Stehling L. C. , Strauss R. G. , Toy P. T. C. Y.
Published: March 2009

Abstract. Because autologous donation is permitted for donors who do not meet homologous blood donation standards, referring physicians and blood center personnel may be concerned about autologous donor reactions. Small studies have determined that mild reactions do not occur more frequently, but the incidence of rarer, more serious, moderate and severe reactions is unknown. We therefore studied the frequency of reactions during 10,200 autologous and 219,307 concurrent homologous donations at four blood centers. No significant difference was seen for severe reactions: autologous 0.039% (4/10,200), homologous 0.037% (82/219,307) (p = 0.79); moderate reactions: autologous 0.19% (19/10,200), homologous 0.22% (473/219,307) (p = 0.60) or mild reactions: autologous 2.26% (231/10,200), homologous 2.26% (4946/219,307) (p = 0.98). We conclude moderate and severe donation reactions do not occur more frequently among autologous donors who are preselected by referring physicians and screened by blood center personnel.
Prevalence of Anti‐HCV Antibody in Blood Donors in the Tokyo Area
Authors: Watanabe Junnosuke , Minegishi Kiyoshi , Mitsumori Tomoko , Ishifuji Makiko , Oguchi Takashi , Ueda Masahiro , Tokunaga Eiichi , Tanaka Eiji , Kiyosawa Kendo , Furuta Seiichi , Katayama Tohru , Kuo George , Choo Qui‐Lim , Houghton Michael , Nishioka Kusuya
Published: March 2009

Abstract. Prospective studies of posttransfusion hepatitis carried out in the past decade showed that 18.1% of the blood transfusions resulted in non‐A non‐B hepatitis in Japan. As an approach to the prevention of posttransfusion non‐A non‐B hepatitis (PTNANB), anti‐hepatitis C virus (HCV) postitivity was measured in 2,970 blood donations in the Tokyo area, and in 200 children aged between 6 and 15 years. Thirty‐four cases were anti‐HCV‐positive, showing an overall positivity of 1.14%. None of the 200 children younger than 15 years old were positive. Correlation of anti‐HCV positivity with the serum ALT levels was observed, but by reducing the accepted ALT levels from 35 Karmen Units (KU) down to 25 KU, it is estimated that 62.5% of the observed PTNANB would still have occurred, and 5.1% of the donated blood could not be used for transfusion. On the other hand, it is estimated that the majority of PTNANB could be prevented, with the loss of 1.14% of donated blood units, using the anti‐HCV screening test.
Comparison of IgM and IgG Anti‐A and Anti‐B Levels in Asian, Caucasian and Negro Donors in the North West Thames Region
Authors: Redman Martin , Malde Ranjan , Contreras Marcela
Published: March 2009

Abstract. This study was undertaken to test the widely held belief that higher levels of immune anti‐A and anti‐B are characteristic of Negro and Asian populations with a corresponding increased risk factor for AB0 haemolytic disease of the newborn. Overall, 300 serum samples from male and female Asian, Caucasian and Negro blood donors in the North West Thames Region of groups A, B and 0 were collected. The sera were titrated in microplates against pooled group A1 and pooled group B red blood cells. Although the results show that the highest levels for IgG anti‐A and anti‐B were found in group 0 female Negro donors, statistically these levels are not significantly higher than those of the other group 0 donors tested. We suggest that the potent anti‐A and anti‐B reported by others in Negro and Asian populations may arise from environmental rather than genetic factors.
Acid Treatment of Platelets as a Simple Procedure for Distinguishing Platelet‐Specific Antibodies from Anti‐HLA Antibodies: Comparison with Chloroquine Treatment
Authors: Kurata Yoshiyuki , Oshida Machiko , Take Hironori , Furubayashi Takayasu , Mizutani Hajime , Tomiyama Yoshiaki , Yonezawa Takeshi , Tarui Seiichiro
Published: March 2009

Abstract. The identification of antibodies to platelet‐specific antigens is important for correctly diagnosing neonatal alloimmune thrombocytopenia, posttransfusion purpura and refractoriness due to platelet‐specific antibodies. However, the serologic identification of these platelet‐specific antibodies is complicated by the presence of anti‐HLA antibodies. We examined and compared the diagnostic usefulness of acid‐treated and chloroquine‐treated platelets for the discrimination of platelet‐specific antibodies from anti‐HLA antibodies. The viability of acid‐treated platelets is 83.4%, which is better than that of chloroquine‐treated platelets (52.6%). The antigenicity of HLA class I antigens of acid‐treated platelets was significantly reduced compared with that of PBS‐ or chloroquine‐treated platelets. On the other hand, platelet surface glycoprotein Ib and glycoprotein IIb/IIIa, and platelet‐specific antigens were stable following acid or chloroquine treatment. Chloroquine‐treated platelets were not suitable targets for analysis by immunofluorescence flow cytometry because of nonspecific fluorescence derived from platelet damage. We conclude that acid‐treated platelets are more suitable targets than chloroquine‐treated platelets for screening for platelet‐specific antibodies and also for analyses of the specificity of platelet‐specific antibodies.
Platelet Antibody Binding and Spontaneous Aggregation in 21 Lupus Anticoagulant Patients
Authors: Wiener Henry M. , Vardinon Nurit , Yust Israel
Published: March 2009

In order to ascertain the role of the antiphospolipid antibody in the pathogenesis of thrombotic disorders, a study of 21 lupus anticoagulant‐positive patients was done by a number of serologic and functional platelet tests. In immunofluorescent studies, we found that 80% showed a mitochondrial pattern on HEp cells and all patients gave an ubiquitously intense staining of donor platelets. By a microscopic spontaneous aggregation test, all lupus anticoagulant‐positive patients showed accelerated platelet aggregation which was calcium‐independent. The results show that the plasma of lupus anticoagulant‐positive patients contains a platelet‐binding antibody and causes spontaneous agglutination of platelets. These properties may play a role in the thrombogenesis common to the antiphospholipid syndrome.
Prospective Study on the Hepatitis Safety of Intravenous Immunoglobulin, pH 4.25
Authors: Rousell Ralph H. , Budinger Miriam D. , Pirofsky Bernard , Schiff Richard I.
Published: March 2009

Abstract. Recent reports of transmission by intravenous γ‐globulin preparations of non‐A non‐B hepatitis (NANBH), including several cases that progressed to severe liver damage and death, have raised concerns about the safety of intravenous γ‐globulin. However, the problem does not seem to be widespread. To assess this issue, we previously reported the results of liver function tests monitored in 41 patients with primary immunodeficiency treated with intravenous immunoglobulin (IGIV), pH 4.25 over periods ranging from 6 to 15 months. Eighteen of these patients at two of the three centers have now had serial serum glutamic pyruvic transaminase (SGPT) levels performed regularly at intervals of 1–5 weeks while continuing monthly intravenous infusions of nonmodified IGIV, pH 4.25 for an additional 14–26 months. The standard dosage was 400mg per kg body weight IGIV, pH4.25. Six lots of IGIV, pH 4.25 were used. Transient minor SGPT elevations were observed in 5 of the patients on a total of 8 occasions. None of the elevations was considered indicative of NANBH or of any chronic hepatic disease. All patients remained negative for hepatitis B surface antigen throughout the study.
Refrigerated Storage of Washed Red Cells1
Authors: Meryman H. T. , Hornblower M. , Keegan T. , Syring R. , Heaton A. , Mesbah‐Karimi N. , Bross J.
Published: March 2009

Abstract. Red cells washed and stored in a citrate‐phosphate‐glucose‐adenine solution at pH 7.4–7.6 demonstrate excellent maintenance of adenosine triphosphate, elevation of 2,3‐diphosphoglycerate well above normal levels for more than 6 weeks, reduced hemolysis and 24‐hour in vivo survival comparable to that of cells stored in ADSOL. These results can be attributed in part to a chloride shift in which the washout of intracellular chloride is associated with an influx of OH“, which increases intracellular pH and thereby increases the rate of glycolysis. The phosphate functions primarily as a buffer to maintain both extra‐ and intracellular pH. Reducing the effective osmolality of the storage solution reduces hemolysis and improves cell morphology.
Antibody‐Coated Erythrocytes Induce Secretion of Tumor Necrosis Factor by Human Monocytes: A Mechanism for the Production of Fever by Incompatible Transfusions
Authors: Hoffman Maureane
Published: March 2009

Abstract. The transfusion of incompatible red blood cells (RBC) rapidly results in the development of fever by an unknown mechanism. In this study, the human monocyte secretion of the endogenous pyrogens tumor necrosis factor (TNF) and interleukin‐1 (IL‐1) was measured. Exposure to human RBC sensitized with antibodies to D or Fya antigens induced the rapid secretion of TNF, but not IL‐1. TNF secretion persisted when monocytes remained in contact with sensitized RBC, but returned to baseline several hours after removal of RBC. Thus, TNF secreted by monocytes may be a mediator of fever during the immune‐mediated destruction of RBC in vivo.
HLA‐DR: Serology versus Restriction Fragment Length Polymorphism
Authors: Nagy M. , Mayr W.R. , Menzel G.R.
Published: March 2009

Abstract. The results of the serological typing and the patterns of restriction fragment length polymorphisms for the detection of HLA‐DR gene products are compared. The data shown demonstrate that the use of DNA typing gives a clearer definition of the HLA‐DR antigens and that the HLA‐DR polymorphism is greater than detected by serology.
Polyreactivity of Human Monoclonal Antibodies: Human Anti‐Rh Monoclonal Antibodies of IgM Isotype Are Frequently Polyreactive
Authors: Blancher A. , Roubinet F. , Oksman F. , Ternynck T. , Broly H. , Chevaleyre J. , Vezon G. , Ducos J.
Published: March 2009

Abstract. The specific aim of this study was to characterize human anti‐Rh monoclonal antibodies cross‐reacting with self‐antigens. We studied supernatants from man‐mouse hybridomas and from lymphoblastoid cell lines. Man‐mouse hybridomas were established by fusion of peripheral blood lymphocytes from healthy individuals recently immunized against Rh alloantigens, with mouse myeloma (or man‐mouse heteromyeloma) cell lines. Lymphoblastoid cell lines were produced by Epstein‐Barr virus induction of lymphocytes from identical sources. Of the 55 monoclonal alloantibodies studied, 11 also reacted with intracellular self‐antigens as demonstrated by immunofluorescence assay on cryostat sections of human tissues. This cross‐reactivity was mainly a property of monoclonal alloantibodies belonging to the IgM isotype (among the 11 cross‐reacting mAbs 10 were IgM). The cross‐reactivities of these monoclonal antibodies were ascertained by absorption of alloreacting antibodies with red blood cells. Similar results were obtained on a panel of purified cellular antigens by ELISA. The results confirm that during an immune response against a foreign antigen (alloantigen), B cells that produce polyreactive antibodies are not excluded from the pool of responding cells. Therefore, polyreactive autoantibodies present in sera from healthy individuals may be the result of an immune response against foreign antigens.
Assignment of the Auberger Red Cell Antigen Polymorphism to the Lutheran Blood Group System: Genetic Justification
Authors: Zelinski Teresa , Kaita Hiroko , Coghlan Gail , Philipps Sylvia
Published: March 2009

Abstract. Family studies have provided the final piece of evidence for the assignment of Auberger to the Lutheran blood group system. Lods derived from combined paternal and maternal meioses (ẑ = 10.83 at ô=0.00) strongly support the contention that a single gene controls the expression of both Au and LU antigens. Consequently, the International Society of Blood Transfusion Working Party on Terminology has designated Aua and Aub as LU18 and LU19, respectively.
High‐Temperature Short‐Time Heat Inactivation of HIV and Other Viruses in Human Blood Plasma
Authors: Charm Stanley E. , Landau Steven , Williams Bolanle , Horowitz Bernard , Prince Alfred M. , Pascual Donna
Published: March 2009

An ultra‐short‐time heating system was used to process blood plasma spiked with various viruses (HIV, vesicular stomatitis virus, encephalomyocarditis virus). Virus reduction and recovery of plasma proteins were measured at various temperatures from 65 to 85 °C. Processing at 77 °C and 0.006 s resulted in a high level of virus kill, including ≥ 4.4 log10 HIV, while maintaining protein structure and activity essentially intact.
Miltenberger Subsystem of the MNSs Blood Group System: Review and Outlook
Authors: Dahr Wolfgang
Published: March 2009

The Miltenberger (Mi) classes represent a group of phenotypes for red cells that carry low frequency antigens associated with the MNSs blood group system. The antigens of this system are known to be located on two sialoglycoproteins denoted as glycophorin A (GP A) and GP B. The structural alterations of seven (classes I, II, III, V, VI, VII, VIII) Mi variants and a related variant (J.L.) have been elucidated. Based on these data and yet incomplete studies of the Mi antigens, the approximate structural alterations in class IV and IX may be predicted. In addition, knowledge of the various structures and partial characterization of the Mi antigens allows one to propose detailed hypotheses concerning the epitopes recognized by the various antibodies that define the Mi subsystem. The understanding of the Mi subsystem at the molecular level paves the way for future studies aimed at a more detailed elucidation of epitopes of Mi‐related antibodies, the characterization of novel Mi variants and a search for hypothetical, hitherto unknown Mi‐related antibodies.
Posttransfusion Hepatitis in Japan
Authors: Takano Susumu , Omata Masuo , Ohio Masao , Satomura Yoichi
Published: March 2009

The incidence of posttransfusion hepatitis and the rate of chronicity were investigated in a program devised at our hospital in December, 1982. Out of 2,596 blood recipients between January, 1982, and December, 1987, 451 (22.7%) developed posttransfusion hepatitis. Seventy‐seven patients out of 217 (35.3%) whose course was closely followed progressed to chronicity. The incidence of posttransfusion hepatitis increased with the volume of transfused blood without any evident limitation. Recipients of elevated‐ALT donor blood (> 26 Karmen units) were found to be more susceptible to posttransfusion hepatitis than those who had received only normal‐ALT donor blood. Packed red blood cells, whole blood and fresh whole blood were high‐risk components, and fresh frozen plasma a low‐risk component of blood. The carrier rate of non‐A, non‐B hepatitis agents in Japanese healthy blood donors was determined to be 1.2% using the Frost‐Reed model of infectious diseases. Anti‐hepatitis C virus was detected in 62% of the cases of posttransfusion hepatitis 1 year after transfusion.
Serological and Chemical Specificities of Twelve Monoclonal Anti‐Lea and Anti‐Leb Antibodies
Authors: Good A. H. , Yau O. , Lamontagne L. R. , Oriol R.
Published: March 2009

The serological specificities of twelve hybridomas were compared as to their chemical reactivity as determined using direct binding to synthetic carbohydrate structures. All anti‐Lea cross‐react with type‐1‐precursor structures and three different variants of anti‐Lea could be defined by their binding to type‐3‐precursor chains, sialylated compounds and the monosaccharide D‐galactose. Three major reactivity patterns were also identified among anti‐Leb reagents. Anti‐LebL cross‐react with Lea and do not significantly bind to H‐related structures. Anti‐LebH L had both anti‐LebL‐like activity (cross‐reaction with Lea) and anti‐LebH‐like activity (cross‐reaction with H). Finally, anti‐LebH cross‐reacts strongly with H compounds and do not bind to Lea. The binding pattern of anti‐LebL suggests that these antibodies have lower affinity for ALeb and BLeb pentasaccharides than anti‐LebH. All these specificities are not absolute, but rather are expressed as members of a quantitative progressive varying series, sugesting the existence of a whole range of antibody specificities gradually changing from Lea → Lea, b → LebL → LebH, L → LebH. The results suggest that anti‐LebL will always cross‐react with Lea and that anti‐LebH will always cross‐react with H related structures. However, under certain well‐defined conditions these cross‐reactions may not be apparent and antibodies might behave as specific anti‐Lea or anti‐Leb in certain tests.
Antiglobulin Testing for CR1‐Related (Knops/McCoy/Swain‐Langley/York) Blood Group Antigens: Negative and Weak Reactions Are Caused by Variable Expression of CR1
Authors: Moulds J. M. , Moulds J. J. , Brown M. , Atkinson J. P.
Published: March 2009

The Knops, McCoy, Swain‐Langley and York antigens have recently been identified as being on complement receptor type 1 (CR1, CD35, C3b/C4b receptor). We examined the relationship between CR1 expression and the reactivity of the CR1‐related blood group antigens with their specific antibodies. RBC from donors of selected phenotypes were tested by hemagglutination using two monoclonal antibodies to CR1, as well as anti‐Kna, ‐McCa, ‐S1a, ‐‘Kn/McC’ and ‐Yka. Monoclonal antibodies 3D9 and Ell required ˜250 and ˜400 CR1/RBC to obtain a positive reaction. Agglutination of antigen‐positive cells by human polyclonal antisera was related to the CR1/RBC: thus, cells expressing 20–100 CR1/RBC were negative and included the previously designated null phenotypes for this collection, 100–150 were weak or negative, and > 200 were usually positive. One RBC sample carried Yka on the 190,000 dalton (A or F allele), but not the 220,000 dalton (B or S allele) variant of CR1, and gave inconsistent reactions with Yka antisera. These data provide an explanation for certain of the serologic characteristics of the CRl‐related blood group antigen system.
Studies in Red Blood Cell Preservation
Authors: Dumaswala U.J. , Petrosky T.L. , Greenwalt T.J.
Published: March 2009

The purpose of this study was to examine whether vesiculation of RBC plays a significant role in their rejuvenation. Outdated units of Adsol® blood, were divided into two aliquots and incubated with equal volumes of a solution of 100 mM pyruvate and inosine, 103 mM phosphate and 5 mM adenine (PIPA) or 0.9% saline. Following 1 h incubation, vesicles were isolated from the supernatants and quantitated for hemoglobin content. Restoration of RBC ATP, 2,3‐DPG, morphology, and osmotic fragility after rejuvenation was satisfactory. The postrejuvenation mean corpuscular volumes (88.2+6.9 fl) were significantly lower (p<0.001) than the prerejuvenation (94.6+6.8 fl) and control (104.0+7.3 fl) volumes. The hemoglobin shed in vesicles during rejuvenation was significantly greater than in the saline controls (0.44+0.31 vs. 0.18+0.10 mg/dl RBCs; p = 0.026). These data suggest that the decreased MCV following rejuvenation is in part due to membrane loss in exocytic vesiculation.
Severe Haemolytic Disease in an Infant Born to an Rhnull Proposita
Authors: Lubenko A. , Contreras M. , Portugal C.L. , Cervi P. , Forsi L. , Garner S.F. , Pinho M.O. , Barbosa M. , Mettrau E.C. , de Azevedo J.G.
Published: March 2009

A 35‐year‐old Brazilian woman (gravida 4, para 2) was delivered of a severely anaemic child whose cord red blood cells had a strongly positive direct antiglobulin test and who required two exchange transfusions within 24 h of birth. Because of the emergency of the situation and the lack of a local immunohaematology reference laboratory, the phenotype of the mother and the specificity of the relevant antibody could not be determined. Hence, compatible blood was not immediately available and the infant had to be given repeated exchange transfusions with incompatible group 0 Rh‐negative blood. The infant is now healthy and thriving.

The mother's red cells were subsequently found to lack all the antigens of the Rh system, and her serum reacted with all red cell samples except those of two unrelated Rhnull individuals. Her serum gave high titres (i.e. 1,024–4,096) by the indirect antiglobulin test against red cells of normal Rh phenotype, as well as against cells with partially deleted Rh phenotypes (titres = 128–512 with –D–·/–D– and ·D·/·D· samples, respectively), and was extremely active in antibody‐dependent cell‐mediated cytotoxicity and monocyte monolayer assays against red cells of normal Rh phenotypes.
An Enzyme‐Linked Antiglobulin Test to Detect Red Cell Globulins after Glutaraldehyde Fixation
Authors: Greenwalt T.J. , Dumaswala U.J. , Siongco A. , Domino M.M.
Published: March 2009

The purpose of this study was to develop an enzyme‐linked antiglobulin test (ELAT) for IgG on RBC without the hemolysis caused by the high pH of the alkaline phosphatase reaction. This was achieved by fixing the RBC with 0.05% glutaraldehyde after attachment of the antibodies. Assays using anti‐D reference standards demonstrated the sensitivity to be 1–2 ng of antibody as compared to 7.5 ng for the manual indirect antiglobulin test. The coefficients of variation of these assays ranged from 10.4 to 20.1%. The mean background absorbance at 405 nm of 105 normal RBC samples was 0.08+0.03 SD. There was an increase in sensitivity of the test after the fixed RBC were stored. Dilute glutaraldehyde stabilizes the RBC membrane and the antigen‐antibody linkage resulting in a more sensitive ELAT.
MINY: A Novel MNS‐Related Blood Group Antigen
Authors: Reid M. E. , Poole J. , Green C. , Neill G. , Banks J.
Published: March 2009

We report an antibody (anti‐MINY) that recognises a novel low‐incidence MNS‐related blood group antigen. Anti‐MINY agglutinates all Hil‐positive red cells tested (Mi.III, Mi.V, Mi.VI, GP.Kipp, GP.Mor and AG) and Hil‐negative, TSEN‐positive red cells (Mi.IV, JR, JL, Oca. and Rag.). All MINY‐positive red cells possess glycophorin A‐B hybrid molecules. The MINY antigen occurs at the unique amino acid sequence which results from the junction of glycophorin A58 to glycophorin B27 regardless of whether the glycophorin B gene encodes methionine or threonine at amino acid residue 29 of normal glycophorin B. The MINY antigen has been provisionally assigned the MNS blood group system number 002.034 on behalf of the ISBT Working Party on Terminology for Red Cell Surface Antigens.
Determination of Total Protein in Highly Purified Factor IX Concentrates
Authors: Löf Anna‐Lena , Gustafsson Georg , Novak Valentina , Engman Lena , Mikaelsson Marianne
Published: March 2009

Protein determination by the methods of Kjeldahl, Biuret, Bradford and UV absorbance at 280 nm have been studied in regard to accuracy, precision and simplicity. A reference preparation of a highly purified factor IX concentrate, Nanotiv, reconstituted to 1/5 of ordinary volume was used in the study in order to make a comparison between the different procedures. The Kjeldahl method resulted in a protein concentration of 3.7 mg/ml, whereas the Biuret, Bradford (BSA) and UV absorbance at 280 nm resulted in protein concentrations of 3.6, 2.5 and 2.8 mg/ml, respectively. The corresponding values for specific activity were 136, 140, 200 and 179 IU/mg, respectively. These results demonstrate a great variation in the response obtained by different methods for determination of total protein.
Acidified Chloroquine Treatment for the Removal of Class I HLA Antigens
Authors: Srivastav Alok , Pearson Helen , Bryant Jennifer , Favaloro Emmanuel , Coulits Nelly , Jindra Jan , Wylie Brenton
Published: March 2009

With the aim of reducing the damage to platelets while effectively removing class I HLA antigens from their surfaces, we developed a new method using acidified chloroquine diphosphate. Platelets were treated with a 0.2 M solution of chloroquine diphosphate (pH 4.0). More than 90% of the platelets remained viable after treatment. While a marked reduction in reactions of acidified chloroquine‐treated platelets with multispecific HLA antisera was noted in comparison with phosphate‐buffered‐saline‐(PBS)‐treated platelets, reactions with platelet‐specific antibodies were preserved. This was demonstrated by immunofluorescence tests and solid‐phase and monoclonal antibody immobilization of platelet antigen assays. Aggregation responses, though reduced in comparison with PBS‐treated platelets, were still preserved after acidified chloroquine treatment. Ultrastructural analysis did not show any significant difference from PBS‐treated platelets. We conclude that treatment of platelets with acidified chloroquine diphosphate is a simple and effective method for removing class I HLA antigens from their surfaces with minimal damage to their structure and function.
Plasma‐Depleted Platelet Concentrates Prepared with a New Washing Solution
Authors: Shimizu Tetsuo , Shibata Kouji , Kora Shin‐ichi
Published: March 2009

In certain clinical situations, complete removal of the plasma proteins from the platelet concentrates (PCs) is necessary by washing prior to transfusion. A simple electrolyte solution with a pH of 6.5 was developed for washing PCs. The platelet‐rich plasma collected with acid‐citrate‐dextrose solution by apheresis in a 0.6‐liter polyolefin bag was centrifuged. After removal of the supernatant plasma from pelleted platelet buttons, 200 ml of a washing solution consisting of 90 mM NaCl, 5 mM KCl, 3 mM MgCl2, 17 mM NaH2PO4, 8 mM Na2HPO4, 23 mM Na acetate, 17 mM Na3 citrate, 23.5 mM glucose, 2 mM adenine, 0.1% dextran, and 28.8 mM maltose (pH 6.5) was added to the pelleted platelet button. Steam sterilization of the solution was carried out under nitrogen to avoid caramelization of glucose. After resuspension of the pelleted platelet button with a washing solution and a second centrifugation, Seto additive solution (Seto sol, pH 7.4) was introduced into the bag to resuspend the platelet buttons for storage for 3 days at 22 °C. All of these procedures were completed within 3 h using a sterile docking device. In washed PCs, 99.1% of the plasma was removed and platelet recovery was 96%. The washed PCs were compared for 3 days with plasma‐poor PCs consisting of 11% plasma and 89% Seto solution. There were no significant differences in percent hypotonic shock response, aggregation, energy metabolism, and morphology of platelets between the two groups during 3 days, except for significant swelling of 3‐day‐old platelets in washed PCs. We conclude that washing PCs with a newly developed medium and resuspension by Seto sol (pH 7.4) had minimum detrimenal effects on further storage for 3 days. It is indicated that the procedure described here is relatively simple and can be finished within 3 h after the collection of platelets, and since plasma removal was sufficiently high, a new plasma‐depleted PC product may be a valuable therapeutic tool for hemostasis therapy with reduced side effects from plasma proteins.
Blood Donations Indeterminate in HIV‐1 Western Blot Analysed by IgM Immunoblot and Polymerase Chain Reaction
Authors: Hopf H. Knüver , Heinze H. , Lambrecht B. , Mohr H. , Beyer J. , Schmitt H.
Published: March 2009

The presence of IgM antibodies to human immunodeficiency virus 1 (HIV‐1) was investigated in blood donor sera which were indeterminate in anti‐HIV‐1 IgG Western blot testing. In 7 of 173 instances out of approximately 1,000,000 blood donation sera with an isolated anti‐p24 IgG produced an anti‐gp41‐45 IgM immunoblot reaction. Applying polymerase chain reaction (PCR) to 29 indeterminate samples out of approximately 125,000 blood donations it was found that 2 of them were IgM‐positive and also contained HIV‐1‐specific DNA sequences. Eleven months later 1 of these 2 donors was retested and found IgM and PCR negative.
Multicenter Evaluation of Methods for Counting Residual White Cells in Leukocyte‐Depleted Red Blood Cells
Authors: Rebulla Paolo , Dzik Walter H. ,   For the Biomedical Excellence for Safer Transfusion (BEST) Working Party of the International Society of Blood Transfusion
Published: March 2009

Two wet workshops were conducted involving 20 laboratories to determine optimum methods for counting residual white blood cells (WBC) in leukocyte‐depleted red blood cells (RBC). In both studies, calibration samples containing known concentrations of WBC were prepared by diluting 1‐day‐old EDTA blood in RBC virtually free of WBC. Study No. 1 was aimed at determining the lower limit of detection and at examining advantages and disadvantages of two methods based on flow cytometry (FC) and on the Nageotte chamber (NC), respectively. This study showed that the lower detection limits for FC and NC were 0.1 and 1WBC/μl, respectively. Methods based on FC showed less variability at 0.1–1 WBC/μl but were equal in performance to counting procedures based on the NC above 1WBC/μl. We then designed study No. 2 to evaluate three different protocols using the NC. In protocol 1, samples were diluted 1:10 with Plaxan and with Türk's solution; in protocol 2 a mixture of Türk's solution and Zap‐oglobin, a reagent used in manual hemoglobin determinations, was used to dilute the samples 1:5; and protocol 3 involved initial 1:10 dilution of a 1‐ml sample with Plaxan followed by WBC concentration into the original 1‐ml volume by centrifugation. In each participating laboratory, two technologists independently processed the samples and read the results with an NC. Plaxan and Türk's solution gave comparable results. Interobserver variability was not critical to the results. All three NC methods were valid for counting at concentrations of 1 WBC/μl. Protocol 3 showed the greatest precision, allowing the measurement of WBC at concentrations of 0.1–1/μl (30,000–300,000 WBC in a 300‐ml RBC unit) with a coefficient of variation of approximately 50% or less. Based on these results the Biomedical Excellence for Safer Transfusion (BEST) Working Party considers all three NC protocols adequate for the routine evaluation of current leukocyte removal filters, i.e. at levels ≥1 WBC/μl. The evaluation of new and more effective filters and validation of filter performance may require techniques with a greater precision at counts below 1 WBC/μl, such as protocol 3. For the latter purposes FC is also applicable. Because the counting techniques were not validated using stored blood, these conclusions only apply to samples obtained from blood stored for 1 day.
Detection of HIV‐1 DNA in Leukocytes Using a Commercially Available Assay
Authors: Zaaijer H. L. , Cuypers H.T.M. , Reesink H.W. , ten Veen J. H. , Schoon‐Visser S. C. E. , Lelie P. N.
Published: March 2009

Recently, an assay for detection of proviral HIV‐1 DNA in leukocytes became commercially available. This assay (Amplicor HIV‐1 test, Roche Diagnostic Systems) multiplies HIV‐1 DNA up to a detectable level, using the polymerase chain reaction. We studied performance of this assay on 74 samples from HIV‐1‐infected patients and on 41 samples from healthy blood donors. Twice a negative control sample appeared to be erroneously reactive. However, sensitivity and specificity on the patient and donor samples both were 100%. To avoid false‐positive results, we advise to repeat initially reactive samples if no other data confirm HIV‐infection.
Bedside Transfusion Errors
Authors: Baele P.L. , De Bruyere M. , Deneys V. , Dupont E. , Flament J. , Lambermont M. , Latinne D. , Steensens L. , van Camp B. , Waterloos H.
Published: March 2009

The true incidence of bedside transfusion errors, i.e. those happening when blood products have left the blood bank, is underestimated because published figures rely on reporting of clinically relevant events or on indirect methods. The SAnGUIS project assessing blood practice in a prospective and randomized fashion for 6 elective surgical procedures gave the opportunity to trace all transfused units and to identify steps at risk during blood delivery in surgery. We considered transfusion of a wrong unit as a major error and poor execution or documentation as a recording error.

Over 15 months, 808 patients out of 1,448 were transfused with 3,485 units. A total of 165 errors were found after blood products had left the blood banks. Seven were misidentifications (0.74% of patients, 0.2% of units). Eight other major errors occurred in 4 (0.5%) patients. Major errors occurred during nonemergency situations, in wards or intensive care units. The remaining (‘recording’) 150 errors consisted of misrecordings (61), mislabellings (6), or failures to document transfusions in the medical records (83).

All errors were uneventful except one misidentification which induced a transient, yet unreported, reaction.

The ‘descending’ inquiry method used for this study showed that most errors pass unnoticed and are therefore not reported. Measurement of error rates may constitute an important quality indicator.

Retrospective information of this survey to the concerned staff people provided an impetus to take adequate measures to reduce these bedside errors.
Evaluation of Third‐Generation Screening and Confirmatory Assays for HCV Antibodies
Authors: Uyttendaele S. , Claeys H. , Mertens W. , Verhaert H. , Vermylen C.
Published: March 2009

A third‐generation (gen.) screening and immunoblot assay (Ortho EIA‐3.0; Chiron RIBA‐3 prototype), using antigens derived from the capsid and different nonstructural regions (NS3, NS4 and NS5) of the hepatitis C virus viral genome, were evaluated in comparison with the corresponding second‐gen. assays (Ortho EIA‐2.0; revised Ortho EIA‐2.5; Chiron RIBA‐2). In 203 depository sera of blood donors, positive in EIA‐2.0, specificity of the screening assays was improved as shown by an increase in positive predictive value for viral carrier state from 0.23 (EIA‐2.0) to 0.37 (EIA‐2.5) and 0.52 (EIA‐3.0). Comparing the confirmation patterns on RIBA‐2 and RIBA‐3, this amelioration was mainly due to the specific elimination of false‐positive c22‐3 and c100‐3 reactions. Antibody response to the newly added NS5 antigen was not as prevalent as to the other antigens and had only a minor influence in sample allocation. In contrast, screening of 1,560 volunteer blood donors and 47 hemodialysis patients revealed 3 additional positive sera, only reacting with the NS5 antigen. However none of these isolated NS5 reactions could be confirmed on synthetic peptides [INNO‐LIA: NS5(p)] and none was PCR positive. A documented seroconversion, detected earlier with EIA‐3.0, was related to a better immunological response to the NS3 antigen and not to the additional NS5. From this pilot study third‐gen. assays appeared extremely useful in the reevaluation of HCV‐seropositive depository sera. However the additional value of the NS5 antigen in blood donor screening is still hypothetical and remains to be established in larger screening studies.
Presence of Anti‐Yta Antibody in a Yt(a+) Patient
Authors: Mazzi Gustavo , Raineri Antonino , Santarossa Liliana , De Roia Dina , Orazi Bianca Maria
Published: March 2009

Since 1956 when Cartwright (Yt) was recognized as a new erythrocyte antigenic system, numerous studies about anti‐Yta and anti‐Ytb antibodies have been published. A number of these studies described the laboratory techniques utilized in antibody identification, while others investigated the clinical importance of anti‐Yta, giving variable results. However, most authors agreed upon the homogeneity of expression in this antigenic system. In our study we have described a case regarding 1 Yt(a+) patient with anti‐Yta antibody. Family studies indicated inheritance of a variant/variant expression of the Yta antigen from the father.
Adsorption of Anaphylatoxins and Platelet‐Specific Proteins by Filtration of Platelet Concentrates with a Polyester Leukocyte Reduction Filter
Authors: Shimizu T. , Uchigiri C. , Mizuno S. , Kamiya T. , Kokubo Y.
Published: March 2009

Anaphylatoxins generated during storage of platelet concentrates (PCs) may potentially have side effects on platelet transfusion. We evaluated the anaphylatoxin‐scavenging abilities of white blood cell reduction filters. Among the commercially available filters for PCs, one made with polyester fiber (PL50) dramatically adsorbed C3a and C4a anaphylatoxins to the respective mean level of 1,721–208 ng/ml and 1,240–141 ng/ml in 3‐day‐old PCs. C3a and C4a were measured as the native and des Arg form of each complement by radioimmunoassay. C3a and C4a anaphylatoxins in the supernatant plasma fraction from 3‐day‐old PC again decreased from 1,136 to 114 ng/ml and from 1,086 to 65 ng/ml, respectively. The filter also adsorbed 85% of platelet factor 4 (PF4) and 31% of β‐thromboglobulin (β‐TG), which had been released from platelets into the plasma during storage. The plasma levels of adhesive proteins such as fibronectin, fibrinogen, and von Willebrand factor, and plasma lactate dehydrogenase activity did not decrease after filtration. Another polyester filter (PL5A), on the other hand, significantly increased C3a and C4a levels with filtration. In addition, there was no PF4 adsorption ability during the filtration. The filters for red cells (RC50, BPF4, and R500A) had no anaphylatoxin adsorption capabilities. The observed specific adsorption of anaphylatoxins might be attributed to the electrostatic force between the positively charged anaphylatoxins with high p1 and the possibly negatively charged filter membranes. Since PF4 and β‐TG have positively charged moieties in the C‐terminal position, the same adsorption mechanism might operate. We have obtained a useful scavenging filter for the evaluation of side effects of anaphylatoxins on patients who received old PCs.
Relative Importance of Immune and Non‐Immune Causes of Platelet Refractoriness
Authors: Doughty H. A. , Murphy M. F. , Metcalfe P. , Rohatiner A. Z. S. , Lister T. A. , Waters A. H.
Published: March 2009

In this prospective study, 26 consecutive patients being treated for haematological malignancies receiving standard (i.e. non‐leucocyte‐depleted) blood components were observed for the development of refractoriness to platelet transfusions. One hundred and sixteen of the 266 (44%) platelet transfusions failed to produce a satisfactory response. In 102/116 (88%), the poor response was in the presence of non‐immune factors known to be associated with platelet refractoriness. Non‐immune factors were present alone in 78/116 (67%), and in combination with immune factors in a further 24/116 (21%). Immune factors (HLA and platelet‐specific antibodies) were present during 29/116 (25%) of unsuccessful platelet transfusions. Statistical analysis confirmed that platelet refractoriness was significantly associated with the presence of nonimmune factors. The non‐immune factors associated with refractoriness were often multiple, most frequently a combination of fever, infection and antibiotic therapy. This study provides evidence that immune mechanisms were not the predominant cause of platelet refractoriness in the patient population studied. It also suggests that measures for the prevention of HLA alloimmunisation, such as leucocyte depletion, may have a limited impact in reducing the incidence of refractoriness to platelet transfusions.
Application of the MAIEA Assay to the Kell Blood Group System
Authors: Petty A. C. , Daniels G. L. , Tippett P.
Published: March 2009

The monoclonal antibody‐specific immobilisation of erythrocyte antigens (MAIEA) test has been employed to investigate the Kell system using five monoclonal antibodies which recognise high frequency epitopes on the 93,000‐molecular weight Kell glycoprotein: BRIC 18, BRIC 68, BRIC 107, BRIC 203 and 6–22. BRIC 107, which has anti‐k‐like (KEL2) specificity, identifies a distinct epitope, whilst competitive binding assays suggested that BRIC 203 (anti‐Kpbc), BRIC 18, BRIC 68 and 6–22 (anti K14) comprise an overlapping set of epitopes. The MAIEA assay has been very successful in confirming the assignment of most of the Kell and para‐Kell antigens to the Kell protein. Due to the competitive nature of the assay and the fact that the monoclonal antibodies bind to different regions, the results also suggest the relative positions of some of the Kell antigens on the Kell protein; these appear to be located in at least five spatially distinct regions.
Inactivation and Elimination of Viruses during Preparation of Human Intravenous Immunoglobulin
Authors: Uemura Yahiro , Yang Y.H. Joy , Heldebrant Charles M. , Takechi Kazuo , Yokoyama Kazumasa
Published: March 2009

We report here the results of our evaluation of virus inactivation during the manufacturing steps of two intravenous immunoglobulin (IGIV) preparations. Virus inactivation and/or removal by processing steps, such as ethanol fractionation and polyethylene glycol precipitation, and deliberate virucidal steps, such as solvent/detergent treatment and pasteurization, were tested on a variety of human pathogenic and experimental model viruses, including human immunodeficiency, Hepatitis C, Mumps, Vaccinia, Chikungunya, Vesicular Stomatitis, Sindbis, and ECHO viruses. All viruses were successfully inactivated and/or eliminated by the processing steps studied. In some cases, however, multiple steps were required. We conclude that the incorporation of steps deliberately designed to inactivate or remove viruses during the production of IGIV provides an extra measure of viral safety.