bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Flow‐Cytometric Equivalent of the Kleihauer Test
Authors: Nelson M. , Zarkos K. , Popp H. , Gibson J. 
Published: December 2004

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

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

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

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

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

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

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

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

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

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

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

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

AbstractCreutzfeldt‐Jakob disease (CJD) has been transmitted in the laboratory and also by iatrogenic accident. However, research has failed to find evidence that its most common form (sporadic CJD) is a natural infection and, in particular, that there is a causal link with scrapie. Bovine spongiform encephalopathy (BSE) probably resulted from scrapie infection in cattle food. In the wake of the BSE epidemic, a novel clinico‐pathological form of CJD has been recognized: new variant CJD (nvCJD). This paper reviews the relationship between nvCJD and BSE and presents the accumulated evidence supporting the view that nvCJD resulted from BSE contamination of human food.