bluuebunny/crossref_metadata_2025_split huggingface dataset:
Application of the Human Hepatitis B Virus Core Antigen from Transgenic Tobacco Plants for Serological Diagnosis
Authors: Tsuda Shinya , Yoshioka Keiko , Tanaka Takeshi , Iwata Akiko , Yoshikawa Akira , Watanabe Yuichiro , Okada Yoshimi 
Published: December 2004

Abstract Background and objectives: The aim was to produce HBcAg from plants more cheaply than can be done by other currently available means, and to apply such antigen to immunoassay procedures for pretransfusion testing of donor blood. Materials and methods: Transgenic Nicotiana tabacum cv. SR‐1 plants expressing the human hepatitis B virus (HBV) core antigen (HBcAg) gene were generated by Agrobacterium‐mediated transformation. The recombinant product, called tHBcAg, can assemble itself into a spherical particle with a diameter of 25 to 30 nm, and can maintain two antigenic determinants of HBcAg, namely HBc/α and HBc/β. Partly purified tHBcAg was used in the hemagglutinationinhibition (HI) test, as routinely used by the Japanese Blood Center, to test a panel of 524 blood units taken from HBV‐positive donors. Results: In the HI test, tHBcAg showed serologic properties comparable to that from Escherichia coli, the standard antigen used in the Japanese Blood Center. Conclusions: Transgenic plants can produce reagents for serologic testing and perhaps even such medical materials as oral vaccines.
Alternative Therapies for Neonatal Autoimmune Thrombocytopenia
Authors: Ovali Fahri , Samanci Nedim , Ermiş Bahri , Akdoǧan Zeliha , Daǧoǧlu Türkan 
Published: December 2004

Abstract Objective: This study was performed to test whether corticosteroids were superior to intravenous immunoglobulins (IVIG) in the treatment of neonatal autoimmune thrombocytopenia (NAT). Methods: All cases received IVIG, and unresponsive cases received corticosteroids. Results: Of 7 babies who received IVIG, only 1 responded. The 6 remaining cases received corticosteroids therafter, and all of them responded well to this therapy. Conclusions: Corticosteroids may be more effective than IVIG in NAT.
Severe Neonatal Alloimmune Thrombocytopenia due to Anti‐HPA‐3a
Authors: Boehlen F. , Kaplan C. , de Moerloose P. 
Published: December 2004

Abstract Background and Objectives: Neonatal alloimmune thrombocytopenia is usually attributable to HPA‐la antibodies. We report a case of parental platelet antigen incompatibility associated with a severe neonatal thrombocytopenia secondary to alloimmunization to HPA‐3a. Materials and Methods: Platelet antibodies were detected by the monoclonal antibody‐specific immobilization of platelet antigens, genotyping of the platelets by PCR, and HLA typing by serologic procedures and PCR. Results: Genotyping of maternal and paternal platelets confirmed the incompatibility in the HPA‐3a system. It is noteworthy that the mother is of the HLA type DRB3*0101, is ABO‐incompatible with her husband, and also has HLA class I antibodies. Conclusion: Severe neonatal thrombocytopenia associated with HPA‐3a alloimmunization is infrequent and all the factors mentioned above could have played a role in the severity of the disease.
Kinetics of Bradykinin Levels during and after Leucocyte Filtration of Platelet Concentrates
Authors: Hild M. , Söderström T. , Egberg N , Lundahl J. 
Published: December 2004

Abstract Objective: To measure beadykinin levels in platelet concentrates during and after leucocyte filtration. Methods: Platelet concentrates were leukocyte depleted using three different leucocyte‐depletion filters selected to represent filters with negative, positive or neutral charge, respectively. Bradykinin levels were analysed before, during and up to 90 min post‐filtration. Results: Significant levels of bradykinin were generated when negatively charged leucocyte depletion filters were used. However, we found a high variation between platelet concentrates prepared from different donors as well as within the same concentrate when this was divided into three aliquots. Generated bradykinin was rapidly degraded in the collection bag and no bradykinin was detectable 60 min post‐filtration. Conclusion: Bradykinin generation is more pronounced when negatively charged leucocyte removal filters are used. Due to a rapid degradation of bradykinin in the storage bag, pre‐storage filtration should minimise the risk of bradykinin related adverse reactions during transfusion with some filters.
Use of Overlapping Synthetic Peptides to Characterize Samples from Blood Donors with Indeterminate Results to Hepatitis C Virus Core Antigen
Authors: León Pilar , López José A. , Elola Consuelo , Lee Stephen R. , Calmann Mark , Echevarría José M. 
Published: December 2004

Abstract Background and Objectives: Despite recent improvements in supplemental assays, isolated reactivity to the hepatitis C virus (HCV) core antigen continues as one of the main problems in the confirmation of anti‐HCV in blood donors. Reactivity against individual peptides from the c22‐3 HCV recombinant antigen has been described as a useful tool for anti‐HCV confirmation and donor counseling in such cases. Materials and Methods: We used a previously described set of overlapping peptides spanning the entire sequence of the c22‐3 antigen to study 87 single serum samples from blood donors with reactivity for c22‐3 antigen alone in second generation recombinant immunoblot assay (RIBA‐2). All of them had been previously studied by RIBA‐3, anti‐HCV E2 EIA and HCV PCR. Results: 66 of the 87 samples studied could be classified as positive or negative for anti‐HCV core using the multipeptide assay and such classification correlated well with the results obtained with RIBA‐3 and the anti‐E2 EIA. However, some discrepancies were found. The epitopes located along the N‐terminal half of the molecule were mainly responsible for the specific antibody recognition but those enclosed within amino acids 1–15 were frequently involved in nonspecific reactivity. Some 38% of samples were considered to have specific antibody to the c22‐3 antigen and a further 9% reacted for both anti‐E2 and single core peptides that were often involved in specific antibody recognision. Conclusion: Testing of blood donor samples indeterminate to the HCV c22‐3 antigen for reactivity against individual core peptides can confirm the presence of specific antibody and recognize nonspecific reactivity with certain cross‐reacting epitopes. Third generation supplemental tests have reduced such false reactivity, but confirmation of samples with anticore alone is still necessary. Single reactivity to the HCV core antigen is likely to reflect prior exposure to the virus, but rarely active infection, either acute or chronic.
Hepatitis E Virus: Relevance in Blood Donors and Other 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 an acute self‐limited disease. HEV is associated with feces‐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 of parenteral infections, namely, hemodialysis patients and children infected with hepatitis C virus (HCV) via blood transfusion. Materials and Methods: We investigated the prevalence of anti‐HEV in 863 blood donors, 63 hemodialysis patients, and 42 children infected with HCV posttransfusion. Results: The prevalence rates were 2.8, 6.3%, and 0 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 parenterally to children.
Prevention and Management of Platelet Transfusion Refractoriness
Authors: Novotny V.M.J. 
Published: December 2004

AbstractPlatelet transfusion refractoriness is a major complication of long‐term platelet supportive care. Refractoriness may lead to fatal bleeding complications in thrombocytopenic patients. Major factors involved are factors related to the clinical condition of the patient as well as HLA alloimmunisation. Non‐alloimmune factors may occur in up to 80% of the patients. However, platelet transfusion outcome is impaired in only 50% of the patients having these conditions. HLA alloimmunisation has been convincingly reduced by the use of leucocytedepleted transfusions. UV‐B irradiation of platelet transfusions may be alternatively used to reduce HLA alloimmunisation. Despite these measures, patients with a history of pregnancy or non‐leucocyte‐depleted transfusions form HLA antibodies in a high proportion (up to 50%). HPA antibodies play a minor but relatively important role in patients with HLA antibodies. ABO antibodies may play a role in refractoriness, which can be abolished by transfusion of ABO‐identical platelets. Screening for the presence of HLA and/or HPA antibodies is indicated in case of transfusion failure after ABO‐identical or HLA‐matched platelets. If no alloantibodies are detected, further analysis to define a role of drugrelated or autoantibodies is required. In case of HLA and/or HPA alloimmunisation associated with refractoriness, matched platelet transfusions are indicated. In case of non‐alloimmune factors associated with increased platelet consumption, increasing the transfusion frequency can be considered. Additional investigations are still necessary to define risk factors for secondary HLA alloimmunisation and refractoriness due to non‐immune factors to further decrease the incidence of refractoriness.
Use of Bovine Viral Diarrhoea Virus as an Internal Control for Amplification of Hepatitis C Virus
Authors: Cleland A. , Nettleton P. , Jarvis L.M. , Simmonds P. 
Published: December 2004

Abstract
Background and Objectives:
Screening for hepatitis C virus (HCV) by polymerase chain reaction (PCR) will be mandatory for screening blood and plasma donors in Europe and elsewhere. This study describes an internally controlled, highly sensitive PCR method designed for screening blood donations in pools. Material and Methods: RNA extracted from bovine viral diarrhoea virus (BVDV) was used as an internal control to monitor the efficiency of extraction, reverse transcription and amplification steps in HCV PCR. Results: Sensitivity of PCR for single molecules of HCV in the presence of 33 genome equivalents of BVDV RNA was achieved by reducing the efficiency of BVDV amplification. BVDV could be recovered at high efficiency from large volume pools (2–5 ml) by ultracentrifugation and by the NucliSens extraction method. Conclusion: Detection of BVDV validates the extraction, reverse transcription and amplification methods used for HCV detection in plasma pools and provides valuable quality assurance for negative results.
Quantitating Donor Behaviour to Model the Effect of Changes in Donor Management on Sufficiency in the Blood Service
Authors: Whyte Gordon 
Published: December 2004

Abstract
Background and Objective:
To describe donor behaviour quantitatively and apply the information on actual donations derived during observed changes in blood collection in Melbourne between July 1990 and June 1996 to construct a logical model to examine the effects of changes in donor management on the sufficiency of the blood supply. Method: Computerised donation data files were searched to determine time to next or previous donation for donors giving blood between 1990 and 1996 stratified by age, donation history and venue. Actual numbers of whole blood donation given by new and repeat donors, July 1990 to June 1996, and numbers of donors refused were used to construct and test a logical predictive model. Results: 558,682 donation intervals were analysed. Donor return rates at 2 years were shown to increase with donor donation history and donor age. The prediction model showed that a 25% decline in whole blood collections over 2 years could be explained by the cumulative effect of a decrease in donor return rates of 2–4%. Between 1990 and 1996 many donors moved from static city collection sites to suburban mobiles. New donor attendance correlated with repeat donor attendance. Donor complaints correlated with donor deferral numbers. Conclusions: The model showed that large shifts in nett blood collections can be explained by relatively small shifts in donor return rates at 2 years.
Effect of Heat on Stored Red Cells during Non‐Flow Conditions in a Blood‐Warming Device
Authors: Eastlund Ted , Van Duren Albert , Clay Mary E. 
Published: December 2004

Abstract
Background and Objectives:
While blood is flowing within a transfusion‐warming device, the blood temperature is usually less than that applied externally. If the flow is temporarily stopped, the temperature can rise above 37°C in some warming devices. We sought to determine whether temperatures near 45°C achieved during prolonged non‐flow conditions in a blood warmer are harmful to red cell integrity. Materials and Methods: After 42 days of storage at 4°C, red cells were exposed to 44.7°C for 30 min while stationary in a blood warming device (Augustine Medical, Inc., 241 Fluid Warming Set) and examined for cell counts, hemolysis and osmotic fragility. Results: Red cell, white cell and platelet counts, hemoglobin, PCV and potassium were unchanged following heat treatment. Plasma hemoglobin was 508±132 mg/dl following heat treatment compared to 396±188 for the control (p>0.05). In the osmotic fragility test, hemolysis remained within normal limits when tested at 0.60 and 0.65% sodium chloride (NaCl) and was unchanged at the 0.5% NaCl level. At the 0.75% NaCl level, there was 16±5.1% hemolysis of heated 42‐day‐old red cells compared to 11±3.4% for the control (both of which being above the 9% upper limit for fresh control red cells). Conclusions: We conclude that elevated temperatures achieved during temporary cessation of flow in the Augustine Medical, Inc., 241 Fluid Warming Set for as long as 30 min do not cause notable hemolysis or other damage to red cells.
Reduction of Platelet Transfusion Associated Sepsis by Short‐Term Bacterial Culture
Authors: Liu Hing‐wing , Yuen Kwok‐yung , Cheng Tammy Shui‐ying , Lee Kwan‐bun , Chua Elizabeth Kin‐ming , Ho Pak‐leung , Lin Che‐kit 
Published: December 2004

Abstract
Background and Objectives:
There is as yet no suitable routine laboratory test for a blood transfusion service to detect bacterial contamination in platelets. This study evaluates the effectiveness and the applicability of short‐term bacterial culture for such a purpose. Materials and Methods: Samples from 5‐unit platelet pools were inoculated into an aerobic culture bottle, then monitored for 48 h at 35°C in an automated monitoring and detection system. Results: 26,210 whole‐blood‐derived platelet components were tested, of which 14 (0.053%) platelet units were found to be contaminated. In addition, nine of the associated red cell units and 4 fresh‐frozen plasma units grew the same organisms on culture. Conclusion: Short‐duration bacterial culture by an automated system is effective and suitable for routine screening in a regional transfusion center.
Transfusion‐Associated Hepatitis in a Tertiary Referral Hospital in India
Authors: Saxena R. , Thakur V. , Sood B. , Guptan R.C. , Gururaja S. , Sarin S.K. 
Published: December 2004

Abstract
Background and Objectives:
In Indian blood banks, screening for hepatitis B virus (HBV) is currently done by the EIA method, but no routine screening is done for hepatitis C virus (HCV). Materials and Methods: To determine the incidence of transfusion‐associated HCV hepatitis, and of any residual transfusion‐associated hepatitis (TAH) after HBsAg screening, we prospectively studied 182 patients who underwent surgery and received blood transfusion. These recipients had normal alanine aminotransferase (ALT) and were negative for HBsAg (monoclonal EIA), and anti‐HCV (third‐generation EIA) before receiving transfusion. Results: Of the 818 blood units transfused after routine screening (average 4.49±3.3 U/patient, range 1–14), 14 (1.7% of units) were found to be infected. Of the 182 recipients, 14 (7.69%) developed TAH during a follow‐up of 6 months, 3 (21.4%) from HBV, 10 (71.5%) from HCV, and 1 (1.7%) from a coinfection of HBV and HCV. All patients with TAH due to HCV were asymptomatic. One patient with TAH due to HBV (33%) and 5 with TAH due to HCV (50%) developed chronic infection with persistently elevated ALT at 6 months. Conclusions: With the current screening practices, the incidence of TAH remains high in India and is mainly due to HCV infection. Furthermore, the screening methods for HBV also need to be improved.
HPA Genotyping by PCR‐SSP: Report of 4 Exercises
Authors: Metcalfe P. , Cavanagh G. , Hurd C. , Ouwehand W.H. 
Published: December 2004

Abstract
Background and Objectives:
Polymerase chain reaction with sequence specific primers (PCR‐SSP) is widely used for the determination of the alleles encoding the human platelet antigens (HPA)‐1 to 5. In order to evaluate and improve performance with this technique, four exercises were organised during 1996‐1998. Materials and Methods: Coded DNA samples were distributed from the National Institute for Biological Standards and Control (NIBSC) as follows: exercise one, 18 samples; two, 12 samples; three, 6 samples, and four, 4 samples. Results: Performance improved over the four exercises following the adoption of a consensus protocol and the re‐design of one primer. The percentage of incorrect results in each exercise was as follows: exercise one, 9%; two 3.2%, three, 0.8%, and four, 0.3% Conclusion: The modified PCR‐SSP protocol is a reliable method for genotyping HPA‐1 to 5 in reference laboratories. DNA‐based HPA genotyping has an important role in platelet immunology and further exercises will be included in the bi‐annual platelet immunology exercises organised by NIBSC.
Application of a Time‐Resolved Fluoroimmunoassay for the Analysis of Normal Prion Protein in Human Blood and Its Components
Authors: MacGregor Ian , Hope James , Barnard Geoff , Kirby Louise , Drummond Olive , Pepper Duncan , Hornsey Valerie , Barclay Robin , Bessos Hagop , Turner Marc , Prowse Christopher 
Published: December 2004

Abstract
Background and Objectives:
To quantify the cellular isoform of prion protein (PrPc) in human blood using a new time‐resolved dissociation‐enhanced fluoroimmunoas‐say (DELFIA®). Materials and Methods: The DELFIA was optimised for human blood samples and applied to isolated cell and plasma fractions from blood donations. The physicochemical properties of PrPc were analysed. Results: 26.5% of blood PrPc was associated with the platelet fraction, 0.8% with polymorphonuclear leucocytes, 2.4% with mononuclear leucocytes, 1.8% with red cells and 68.5% with plasma (mean values from 4 processed donations). Conclusion: The majority of blood PrPc is found in the platelet and plasma compartments.
A New Method for Phenotyping Red Blood Cells Using Microplates
Authors: Llopis F. , Carbonell‐Uberos F. , Montero M.C. , Bonanad S. , Planelles M.D. , Plasencia I. , Riol C. , Planells T. , Carrillo C. , De Miguel A. 
Published: December 2004

Abstract
Background and Objectives:
The supply of phenotyped red blood cells (RBC) for patients with several RBC antibodies presents a difficult task to hospital blood banks and regional blood centers. The aim of this study was to establish a low‐cost typing system to allow extensive phenotyping of regular blood donors for clinically significant RBC antigens. Materials and Methods: We developed a new buffer that greatly intensifies the antigen‐antibody reaction and thus reduces the quantity of serum needed for phenotyping. The procedure was carried out on microplates. Results: A total of 20,435 regular blood donors have been typed to date. For 752 units required for transfusion, 3,584 phenotyping tests were performed, validating the results by tube or gel typing methods; agreement was achieved in all cases. Conclusion: This technique seems adequate for phenotyping a large number of RBC units at very low cost, thus facilitating the availability of phenotyped blood.
The Direct Antiglobulin Test: Still a Place for the Tube Technique?
Authors: Tissot Jean‐Daniel , Kiener Cèline , Burnand Bernard , Schneider Philippe 
Published: December 2004

Abstract
Objectives:
Antibodies of different immunoglobulin isotypes, or complement, may coat red blood cells in vivo. They are detected by the direct antiglobulin test (DAT), usually performed by the conventional tube technique. The purpose of this study was to compare the latter technique with the gel test. Methods: Three hundred and ninety‐eight consecutive samples, sent to our laboratory for direct antiglobulin testing, were analyzed with the tube technique and the gel test, using reagents from different manufacturers. Eighty‐seven samples had been collected from newborns and 23 from fetuses. Results were expressed as positive or negative. Results: In 162 out of 398 cases, the DAT was negative with both methods, whereas in 178 out of 398 cases, the DAT was positive with both techniques using polyspecific antibodies (observed agreement: 84.5%; x = 0.71). Discrepancies between the two methods were observed in 58 cases: 51 samples appeared as DAT positive using the tube method and negative with the gel test, whereas only 7 were positive exclusively with the gel test. Among the 178 samples that were positive with both techniques, 93 samples showed discordant results when evaluated with anti‐lgG and 82 with anti‐C3d, respectively). The sensitivity of the DAT performed by the gel test, in comparison with the conventional tube technique, was 75.4% (95% confidence interval (CI):69.4–80.8).96.8% (95% CI:92.8–99.0), and 16.3% (95% CI: 9.8–24.9) with polyspecific, anti‐IgG and anti‐Cd3 reagents, respectively. Conclusions: The gel test appeared to be less sensitive than the conventional tube technique when utilized for DAT, particularly when C3d was present on red blood cells. These results emphasize that before implementing a new technique in the laboratory, comparison with existing techniques, using different reagents, is mandatory.
Retention of Coagulation Factors in Plasma Frozen after Extended Holding at 1–6°C
Authors: Smith Janis F. , Ness Paul M. , Moroff Gary , Luban Naomi L.C. 
Published: December 2004

Abstract
Background and Objectives:
The ability to use plasma, isolated form units of whole blood and frozen within 24 h of phlebotomy, as a substitute for plasma forzen within 8 h of phlebotomy would have several advantages for blood centers. It should provide increased flexibility pertaining to the freezing of plasma for clinical use. We have conducted studies to assess the influence of an extended holding time for separated plasma, prior to freezing, on the retention of coagulation factor activity. Study Design and Methods: Freshly harvested plasma from each of 10 units of CPD‐whole blood was divided into four equal aliquots. These aliquots were held in plastic packs at 1–6°C for a total of 0, 8,15 and 24 h. Susequently, the plasma aliquots were forzen rapidly and stored at −20°C for 4 months. The thawed plasma was tested for coagulant factors V and IX, factor VIII coagulant activity (factor VIII:C), von Willebrand factor antigen (vWF:Ag) and ristocetin cofacotor of von Willebrand factor. Results: The levels of factor V, factor vWF: Ag, factor IX and ristocetin cofactor were not influenced by holding the plasma for up to 24 h prior to freezing. Factor VIII:C activity was reduced with extended holding at 1–6°C; the percentage at time zero activity was 75.9±2.4% for samples forzen immediately after a 24‐hour period. Conclusions: The data indicate that coagulation factor properties of harvested plasma are retained except for factor VIII for at least 24 h prior to freezing.