bluuebunny/crossref_metadata_2025 huggingface dataset:
Laboratory diagnosis and molecular epidemiology of an outbreak of hepatitis C virus infection among recipients of human intravenous immunoglobulin in Spain
Authors: Echevarría J. M. , León P. , Domingo C. J. , López J. A. , Elola C. , Madurga M. , Salmerón F. , Yap P. L. , Daub J. , Simmonds P.
Published: March 2003

BACKGROUND: Passive transfer of antibody to hepatitis C virus (HCV) has been thought to occur after infusion of human intravenous immunoglobulin (IVIG), as anti‐HCV and/or HCV RNA was commonly found in that product. Recently, however, HCV RNA was detected in the serum of recipients of IVIG. Establishment of a causal relationship between IVIG therapy and HCV infection in recipients was attempted. STUDY DESIGN AND METHODS: Anti‐HCV and HCV RNA sequences were investigated in serum samples from 39 persons who received a human IVIG product in seven different hospitals in Spain. HCV RNA was also investigated in two batches of the IVIG shared by some recipients. All the viral RNA detected were characterized with a line probe assay, restriction fragment length polymorphism analysis of the 5′‐noncoding and core regions, and sequencing of the nonstructural 5 region. RESULTS: On the basis of both clinical and laboratory data, a relationship could be established between the IVIG therapy and the acquisition of the HCV infection by the recipients. Several HCV strains were detected among the recipients, with most of the recipients coming from the same hospital presenting with closely related strains. Moreover, an HCV strain almost identical to the main strain detected among the recipients was found in one batch of the IVIG that probably was shared by most of them. Follow‐up studies and evaluation of low‐avidity anti‐ HCV IgG suggested that both acute primary infections and reinfections were produced. In one case, direct evidence of reinfection by a different HCV strain was obtained. CONCLUSION: The results did not exclude the possibility that a second HCV strain associated with a further, unidentified batch of the IVIG could have contributed to this outbreak.
Cryopreservation of hematopoietic progenitor cells with 5‐percent dimethyl sulfoxide at −80° C without rate‐controlled freezing
Authors: Galmés A. , Besalduch J. , Bargay J. , Matamoros N. , Durán M.A. , Morey M. , Alvarez F. , Mascaró M.
Published: March 2003

Background: Cryopreservation of hematopoietic cells with the rate‐controlled method is used in the majority of centers. In recent years, there has been a trend toward the simplification of the process.

Study Design and Methods: A simplified method for cryopreservation was developed with 5‐percent dimethyl sulfoxide (DMSO) as the sole cryoprotectant without rate‐controlled freezing. Experiments were done with progressive concentrations of DMSO, ranging from 0 to 10 percent. With DMSO concentrations from 5‐ to 10‐percent, the best recovery and viability for hematopoietic progenitor cells were observed. Hematopoietic progenitor cells with plasma and 5‐percent DMSO were frozen and stored in a −80° C mechanical freezer. Ten patients with solid and hematologic malignancies underwent transplantation with autologous hematopoietic progenitor cells.

Results: The median number of transfused mononuclear cells and CD34+ cells was 3.70 (3.1–8.2) × 108 per kg and 1.70 (0.8‐6.5) × 106 per kg, respectively. The median number of transfused colony‐forming units‐granulocyte‐macrophage was 12.45 (3.4–55.3) × 104 per kg. All patients showed rapid and sustained engraftment. The mean times to reach a neutrophil count of 0.5 × 109 per L and a platelet count of 50 × 109 per L were 11.50 ± 1.70 and 13.90 ± 3.98 days, respectively. All patients are alive and without transfusion requirements in complete remission 2 to 8 months after transplantation.

Conclusion: This simplified cryopreservation technique will be useful for institutions without rate‐controlled freezing facilities. Moreover, this method diminishes the amount of DMSO infused to patients, as well as its toxicity.
Allele‐specific restriction analysis of human platelet antigen system 4
Authors: Matsuo K. , Reid D.M.
Published: March 2003

Background: Diagnosis, and occasionally treatment, of disorders involving platelet‐specific alloimmunization, including neonatal alloimmune thrombocytopenia and posttransfusion purpura, requires platelet allotyping. Allele‐specific restriction analyses for convenient genotyping are available for the major human platelet antigen (HPA) systems except HPA‐4.

Study Design and Methods: An allele‐specific restriction analysis for HPA‐4 was developed by designing a polymerase chain reaction primer containing a single‐base substitution (A for T) at glycoprotein IIIa cDNA position 529. The resulting mismatch did not interfere with DNA amplification. Digestion of the polymerase chain reaction product with BsmI allowed differentiation of HPA‐4a and HPA‐4b alleles.

Results: A 126‐bp polymerase chain reaction product was amplified with the novel primer. BsmI endonuclease cleaved product encoding HPA‐4a into 104‐ and 22‐bp fragments and left DNA encoding HPA‐4b intact.

Conclusion: This rapid allele‐specific restriction analysis for genotyping the HPA‐4 system complements similar methods for other platelet alloantigenic determinants.
The KEL24 and KEL14 alleles of the Kell blood group system
Authors: Lee S. , Naime D. , Reid M. , Redman C.
Published: March 2003

BACKGROUND: The Kell blood group system consists of at least 21 antigens, which may be classified into five sets of alleles and at least 10 independently expressed antigens. The molecular basis of four of the five sets of alleles has been described; point mutations in KEL leading to amino acid substitutions characterize the alleles. In this study, the point mutation associated with the remaining allele, KEL14/KEL24, was determined. STUDY DESIGN AND METHODS: The 19 exons of KEL were amplified from genomic DNA by a polymerase chain reaction (PCR) procedure. The PCR products were sequenced. DNA sequences from unrelated KEL:14,24 and KEL:‐14,24 individuals were compared to the DNA sequence of the common KEL:14,‐24 phenotype. RESULTS: DNA from the KEL:14,24 person yielded both G and C at nt 659, indicating an Arg and Pro polymorphism in amino acid residue 180 of Kell protein. DNA from the KEL:‐14,24 person had a G659C mutation in exon 6, indicating an Arg180Pro substitution. The G659C change introduces an Hae III restriction enzyme site, which was used to confirm the base mutations by restriction fragment length polymorphism analysis of the PCR products. CONCLUSION: A G659C mutation, predicting an Arg180Pro change in Kell protein, is associated with the KEL14/KEL24 allele.
Influence of a recombinant hemoglobin solution on blood rheology
Authors: Stetter M.N. , Baerlocher G.M. , Meiselman H.J. , Reinhart W.H.
Published: March 2003

BACKGROUND: Red cell transfusion is a matter of great concern because of viral infections. Recently, a genetically engineered hemoglobin, rHb 1.1, consisting of two alpha chains and one beta chain, has been developed; it has good oxygen‐carrying and ‐unloading capacity and is devoid of renal toxicity. STUDY DESIGN AND METHODS: An in vitro study of the influence of increasing concentrations of rHb 1.1 on plasma and blood viscosity, red cell aggregation and deformability, and neutrophil deformability was performed. RESULTS: The rHb 1.1 (50 g/L in phosphate‐ buffered saline) had a viscosity of 0.80 +/− 0.02 mPa‐sec at 37 degrees C, which was lower than that of normal Hb solution at the same Hb concentration (0.93 +/− 0.01 mPa‐sec, p < 0.001) or of albumin, a protein with similar molecular weight (0.93 +/− 0.01, p < 0.0001). The admixture of rHb 1.1 to plasma or to red cell suspensions, at constant Hb concentration, led to a dose‐dependent decrease in their viscosities. The simulation of replacement therapy during blood loss revealed rheologic properties of rHb 1.1 that were superior to those of all other fluids. The rHb 1.1 did not affect red cell aggregation or the deformability of red cells or white cells, as measured by the cells' transit time through small pores. CONCLUSION: These data indicate that rHb 1.1 has excellent rheologic properties and should hold promise not only as an oxygen‐carrying therapeutic agent, but probably also as a hemodilutional agent that simultaneously decreases blood viscosity and provides oxygen‐carrying capacity.
Cost‐effectiveness of expanded human immunodeficiency virus‐testing protocols for donated blood
Authors: AuBuchon J.P. , Birkmeyer J.D. , Busch M.P.
Published: March 2003

BACKGROUND: This study was designed to estimate the cost‐effectiveness of expanding the human immunodeficiency virus (HIV)‐testing protocol for donated blood beyond screening for HIV antibodies to further reduce the risk of HIV transmission through transfusion.

STUDY DESIGN AND METHODS: A Markov decision analysis model was developed to estimate the cost‐effectiveness of HIV antibody testing (at a cost of $5/unit) and of adding to that protocol a second HIV test, either plasma p24 antigen detection or RNA polymerase chain reaction (PCR) (at costs of $5/unit and $8/unit, respectively). Test efficacy was projected from anticipated window‐period reductions (6 days for p24 antigen, 11 days for RNA PCR), and donor seroconversion rates were derived from the Retrovirus Epidemiology Donor Study.

RESULTS: On the basis of current estimates of HIV prevalence rates in blood donors (1/10,000) and 16 million annual transfusions in the United States HIV antibody testing prevents 1568 cases of transfusion‐acquired HIV infection each year at a cost of $3600 per quality‐adjusted year of life saved. The addition of p24 antigen testing would prevent 8 more cases at a net additional cost of $60 million annually ($2.3 million/quality‐adjusted life year); RNA PCR testing would prevent 16 more cases at a net additional cost of $96 million annually ($2.0 million/ quality‐adjusted life year).

CONCLUSION: Although expanding the donor HIV screening protocol with p24 antigen or RNA PCR testing will prevent rare cases of transfusion‐associated HIV, the cost‐effectiveness of such an addition is predicted to be far below that of most medical interventions. Thus, HIV test protocol additions are unlikely to provide cost‐effective improvements to blood safety in the United States.
Indeterminate human immunodeficiency virus type 1 western blot may indicate an abortive infection in some low‐risk blood donors
Authors: Georgoulias V.A. , Malliaraki N. E. , Theodoropoulou M. , Spanakis E. , Fountouli P. , Tsatsaki D. , Kotsaki S. , Karvela‐Aggelaki A. , Malliaraki‐Pinetidou E.
Published: March 2003

BACKGROUND: The infectious status of persons with an indeterminate human immunodeficiency virus type 1 (HIV‐1) Western blot must be established.

STUDY DESIGN AND METHODS: Evaluation of the CD4 and CD8 T‐ cell subsets and the expression of HIV‐1‐integrated sequences by Southern blot and polymerase chain reaction were studied in a group of low‐risk subjects with an indeterminate Western blot.

RESULTS: From a total of 45,000 blood donors and 50 patients with chronic renal failure on hemodialysis who were tested during the period of 1985 through 1990, 50 sera (0.1%) had an indeterminate Western blot. A low CD4:CD8 ratio (0.7‐1.2) was detected in 14 of 24 tested subjects, whereas the unfractionated and adherence‐enriched cells of 7 (32%) and 5 (23%) of 22 patients, respectively, could be stained with a p24 monoclonal antibody. A transient positive culture was detected in 3 of 20 subjects, but these viral isolates could not be transmitted to CEM‐A310 cells. Ultracentrifuged culture supernatants hybridized under high‐ stringency conditions with genomic gag‐pol (4 cases), env (3 cases), and tat (1 case) cDNA fragments of the HXB2 HIV‐1 clone. In one case, DNA obtained from adherent but not unfractionated mononuclear cells contained 3.3‐ and 3.9‐kb env‐ and gag‐pol‐related HIV‐1 sequences, respectively; these sequences were heavier than expected. Polymerase chain reaction analysis for gag and pol but not env sequences was positive in 1 and 2 of 7 cases, respectively. A female patient with a positive viral culture and who was positive for pol in polymerase chain reaction demonstrated a full seroconversion 19 months later.

CONCLUSION: The results strongly suggest that, rarely, some low‐risk subjects with indeterminate Western blot results might be infected with low‐level replicative strains or HIV‐related viruses; thus, an exhaustive immunologic and virologic workup is needed for the investigation of these subjects.
Inactivation of hepatitis A virus in plasma products by vapor heating
Authors: Barrett P.N. , Meyer H. , Wachtel I. , Eibl J. , Dorner F.
Published: March 2003

BACKGROUND: The transmission of hepatitis A virus (HAV) has been associated with the use of a number of solvent/detergent‐treated factor VIII concentrates and possibly a factor IX concentrate. These reports have emphasized the necessity of using virus‐inactivation methods for plasma products that are capable of inactivating nonenveloped viruses such as HAV. STUDY DESIGN AND METHODS: A simple, highly accurate titration procedure for HAV, which allows extensive kinetic investigations of virus‐inactivation procedures, has been developed. This system has now been used to evaluate the efficacy of vapor heating in inactivating HAV after the addition of the virus to a range of human plasma products. RESULTS: It was demonstrated that HAV was significantly more thermostable than other picornaviruses, which reinforced the fact that such viruses cannot be used as model viruses for HAV‐inactivation studies. A one‐step vapor‐heating procedure was demonstrated to inactivate between 5.9 and > 6.3 log10 of HAV in different products. A two‐step vapor‐heating procedure had the capacity to inactivate between > 8.7 and > 10.4 log10 of HAV. Both procedures were more effective in inactivating HAV than was the pasteurization procedure used for virus inactivation in human albumin solutions. CONCLUSION: These data demonstrate the efficacy of vapor heating in inactivating high‐titer HAV after the spiking of plasma products with virus. This study confirms and explains the results of controlled clinical trials and long‐term clinical usage with respect to the lack of HAV transmission by such vapor‐heated products.
Impact of buffy coat storage on the generation of inflammatory cytokines and platelet activation
Authors: Klüter H. , Schlenke P. , Müller‐Steinhardt M. , Paulsen M. , Kirchner H.
Published: March 2003

BACKGROUND: Whole blood‐derived buffy coat (BC) has become an alternative source from which to prepare random‐donor platelet concentrates. The influence of prolonged storage of BC prior to platelet concentrate preparation is a matter of controversy. The impact of BC storage on cytokine release was evaluated and the platelet activation quantified. STUDY DESIGN AND METHODS: BCs were prepared from whole‐blood donations after hard‐spin centrifugation. After 1, 3, 6, 12, and 24 hours of storage at 22 degrees C without agitation, samples were withdrawn for cell count and blood gas analysis and measurement of interleukin (IL)‐1beta, IL‐6, IL‐8, tumor necrosis factor alpha and platelet factor 4. Platelet surface markers CD41a, CD42b, CD62P, and CD63 were analyzed by flow cytometry, and the antibody‐binding sites were quantified by using microbeads. RESULTS: Inflammatory cytokines IL‐ 1beta, IL‐6, and tumor necrosis factor alpha were hardly detectable in stored BCs but levels of IL‐8 increased in 25 percent of BCs after 24 hours. A constant increase in platelet factor 4 was observed, which accelerated after 12 hours of storage. Analysis of platelet surface markers showed an initial decrease of platelet activation, followed by an increase after storage for 12 to 24 hours. CONCLUSION: Storage of BCs for up to 12 hours without agitation showed a good preservation of platelets but storage of BCs for 24 hours resulted in increased platelet activation and significantly higher release of platelet factor 4 and IL‐8. Stored BCs might well be suitable for platelet preparation, but their storage time should not greatly exceed 12 hours.
A Thr552—>Ile substitution in erythroid band 3 gives rise to the Warrior blood group antigen
Authors: Jarolim P. , Murray J. L. , Rubin H. L. , Coghlan G. , Zelinski T.
Published: March 2003

BACKGROUND: Recent family studies established that the low‐incidence red cell antigen WARR is not part of the MNS, Lutheran, Lewis, Duffy, Kidd, Xg, Chido/ Rodgers, Kx, or Gerbich blood group systems. Continued serologic and genetic studies of WARR suggest that it is carried on erythroid band 3. STUDY DESIGN AND METHODS: To test the hypothesis that expression of WARR is controlled by the anion exchanger 1 gene (AE1), AE1 intronic primers that flank the exons encoding the membrane domain of band 3 were prepared. Polymerase chain reaction‐amplified products corresponding to exons 11–20 of AE1 were analyzed for single‐strand conformational polymorphism (SSCP) in DNA from WARR‐positive and WARR‐ negative individuals. RESULTS: An SSCP was detected in exon 14. Subsequent sequencing revealed a C–>T mutation in codon 552 that leads to a Thr–>Ile substitution. Because the C–>T mutation eliminates a Bbs I restriction site, it was possible to confirm the phenotypes of all family members. To study the possible effect of the Thr552–>Ile substitution on the expression and function of band 3, polymerase chain reaction‐amplified reverse‐transcribed reticulocyte mRNA was digested with Bbs I. Both alleles of band 3 mRNA were detected in similar quantities, which suggests that the substitution underlying WARR did not interfere with mRNA stability. Comparison of sodium dodecyl sulfate‐ polyacrylamide gel electrophoretic mobility and size patterns revealed no difference between proteins isolated from WARR‐positive and WARR‐ negative red cells. Further, the presence of WARR did not alter the di‐ isothiocyano‐dihydrostilbene disulfonate (DIDS)‐inhibitable influx of radiolabeled sulfate. CONCLUSION: Although it appears inconsequential to the function of band 3, the red cell polymorphism known as WARR is controlled by AE1. WARR should be therefore included in the Diego blood group system.
A multicenter evaluation of the routine use of a new white cell‐ reduction apheresis system for collection of platelets
Authors: Fournel J.‐J. , Zingsem J. , Riggert J. , Muylle L. , Müller N. , Köhler M. , Beaumont J.‐L. , Baeten M. , Eckstein R. , van Waeg G.
Published: March 2003

BACKGROUND: Residual white cells (WBCs) cause serious side effects in platelet transfusion. An in‐line WBC‐reduction system based on fluidized particle bed technology was recently developed as a modification of an existing plateletpheresis system.

STUDY DESIGN AND METHODS: In an investigational phase, three flow profiles were evaluated using prototype software in five centers, each using their standard conditions. In the confirmatory phase, the released software was tested in three centers. WBCs were counted in two full Nageotte grids (dilution 1‐in‐5).

RESULTS: With the prototype software, WBC levels were always below 1 × 106 per procedure (median, 25,000/procedure; n = 314). One profile proved to be superior to the other two with respect to platelet yield and residual WBCs, and it was incorporated in the released WBC‐reduction system, together with a built‐in process control. Median residual WBCs in these WBC‐reduction system components not rejected by the process control were 19,000 per procedure (n = 211/225 total), with 99.5 percent of the platelet components having less than 1 × 106 WBCs.

CONCLUSION: The protocol selected in the initial phase, now available as a WBC‐reduction system, results in platelet concentrates with very low residual WBC levels. This satisfies even the most stringent criteria for WBC reduction in platelets, without the platelet loss typically seen with conventional fiber filtration.
Lower antigen site density and weak D immunogenicity cannot be explained by structural genomic abnormalities or regulatory defects of the RHD gene
Authors: Beckers E.A.M. , Faas B.H.W. , Ligthart P. , Overbeeke M.A.M. , von dem Borne A.E.G. , van der Schoot C.E. , van Rhenen D.J.
Published: March 2003

BACKGROUND: The weak D phenotype is characterized serologically by a weak or negative agglutination reaction with polyclonal anti‐D in an immediate‐spin test. Agglutination is enhanced in the indirect antiglobulin test. Red cells that are typed weak D have a much lower number of apparently complete D antigens at their cell surface and are associated with considerably weaker immunogenicity than are red cells with normal D. In a previous study, the number of D sites per cell was determined in eight unrelated weak D individuals to range from 490 to 1870 D sites per cell, which corresponded to 4 to 14.2 percent of the number of D sites in CcDee samples. STUDY DESIGN AND METHODS: The RHD gene was investigated for structural abnormalities by Southern blot experiments and polymerase chain reaction‐based RHD typing in these individuals. In addition, abnormalities in the transcription process were studied by sequence analysis of RH transcripts and by comparing the relative amounts of RHD mRNA in weak D to those in CcDee, CcDEe, and ‐D‐ samples by using a semiquantitative reverse transcriptase‐ polymerase chain reaction analysis. RESULTS: The RHD gene in weak D phenotypes does not show any abnormalities at either the genomic or the transcriptional level when compared to the RHD gene in normal D phenotypes. CONCLUSION: The weaker immunogenicity of weak D is not explained by structural difference in the RHD gene itself. The weaker expression of D might be caused by factors involved in the Rh‐related complex or by an as yet unidentified suppressor gene. This study supports the concept that weak D phenotypes carry complete D polypeptides and reflect a quantitative rather than a qualitative variation of D.
Increased platelet aggregation due to chilling to 20 degrees C is not related to increased sensitivity to agonists
Authors: Hansen Thomas N. , Brockbank Kelvin G.M.
Published: March 2003

BACKGROUND: Previous studies suggested that chilled platelets have a greater sensitivity to agonists than do platelets aggregated at 37 degrees C. STUDY DESIGN AND METHODS: Washed platelets were aggregated at 20 or 37 degrees C with ADP (0‐20 microM), arachidonic acid (0‐200 microM), or the thromboxane mimetic U46619 (0‐9 nM). RESULTS: Chilling caused a significant (p < or=0.05) increase in spontaneous platelet aggregation (> 27% at 20 degrees C vs < 5% at 37 degrees C) and spontaneous dense granule release (> 0.01 nM of ATP at 20 degrees C vs. 0 nM of ATP at 37 degrees C), ADP and U46619 caused a significantly greater aggregation response and dense granule release at 20 degrees C, although there was no change in agonist sensitivity of platelets (effective dose of agonist necessary to induce 50% aggregation [ED50]: 1.00 +/− 0.35 microM ADP at 20 degrees C and 1.63 +/− 0.47 microM ADP at 37 degrees C; 8.26 +/− 3.65 pM U46619 at 20 degrees C and 18.97 +/− 4.82 pM U46619 at 37 degrees C). Platelets aggregated with arachidonic acid showed a significant decrease in aggregation and agonist sensitivity at 20 degrees C (ED50 118.7 +/− 44.4 microM) from those at 37 degrees C (ED50 25.6 +/− 7.2 microM), possibly as a result of the reduced enzymatic activity of cyclooxygenase and thromboxane synthase at the lower temperature. CONCLUSION: The data suggested that washed platelets chilled to 20 degrees C and aggregated are not more sensitive to agonists than are 37 degrees C controls, but rather that chilled platelets undergo greater spontaneous aggregation.
Out‐of‐hospital transfusion
Authors: Evans Cherie S.
Published: March 2003

Although issues regarding the provision of transfusions in nonhospital settings are still unresolved, there is now a considerable body of experience with such transfusions and a growing consensus that there are acceptable ways to make the service available to carefully selected patients. Besides the core of published information on the subject, there is ongoing interest and a comparison of experiences occurring at meetings such as those of the AABB and state organizations such as the California Blood Bank Society. These formal and informal exchanges of information by blood centers, hospitals, and outpatient facilities continue to improve the quality achieved by those just embarking on out‐ of‐hospital transfusion programs and those already experienced in this approach. The ultimate beneficiaries of all this shared quality improvement are those patients for whom high‐quality out‐of‐hospital transfusion is not only appropriate but even preferable to inpatient care.
Donor attitudes about exporting and importing blood
Authors: Bowman R.J. , Clay M.E. , Therkelsen D.J. , Maynard B. , Nass C.C. , Popovsky M.A. , Ryan T.J. , Shafer A. W. , Riedesel P.
Published: March 2003

BACKGROUND: The movement of blood among different areas of the United States and the collection of more blood than is needed locally in some areas are increasing. Little is known of donors' attitudes about this blood resource sharing. STUDY DESIGN AND METHODS: One thousand donors from five regions of the American Red Cross Blood Services were surveyed by telephone. Demographic information about the donors and the regions was obtained, and the donors were asked to describe their attitudes about blood resource sharing as well as other blood donation‐ related issues. RESULTS: Donors are not very knowledgeable about whether their community is self‐sufficient in its blood supply. In regions that import blood, 29 to 43 percent of donors believed that enough blood was collected to meet all local needs, and, in regions that export blood, only 22 to 24 percent of donors believed that more than enough blood was collected. About three‐fourths of the donors believed it acceptable to send their blood to other communities if it is needed there. However, this attitude was based on the premise that all local needs would be met first. Only 4 percent of donors would be less willing to donate if their blood was being sent to another community. CONCLUSION: Donors are not very aware of blood resource sharing but are willing, under certain circumstances, to donate blood for use outside their local communities.
A blood center communication plan and guideline for assisting hospitals with the implementation of the new international uniform bar code‐ labeling standard
Authors: Kerr M. J.
Published: March 2003

BACKGROUND: Transition to the International Society of Blood Transfusion (ISBT) bar code‐labeling standard known as ISBT 128 is scheduled to begin July 1, 1997. Blood centers play a pivotal role in educating and assisting hospitals with the implementation of the new standard. A communication plan and hospital guidelines to facilitate the actual transition to ISBT 128 are integral parts of a blood center's comprehensive implementation strategy. STUDY DESIGN AND METHODS: In Phase 1, the blood center identified specific target groups within the hospitals and developed educational materials for them. A survey tool was used to identify unique hospital needs. Subsequent blood center newsletters and customer service forums included ISBT 128 updates. In Phase 2, guidelines to assist hospitals with the identification of critical elements, milestones, and target dates for the transition were developed with hospital staff input. RESULTS: Survey results on the change to ISBT 128 showed that many hospitals were unfamiliar with this change and its effects on operational issues. Various forums to disseminate information about ISBT 128 are needed to provide education and assistance to diverse groups within the hospital. CONCLUSION: A partnership approach between hospitals and the regional blood center should promote a successful transition from the current bar code symbology, American Blood Commission Codabar, to ISBT 128.
Screening blood donations for viral genomes: multicenter study of real‐ time simulation using pooled samples on the model of hepatitis C virus RNA detection
Authors: Lefrère J‐J. , Coste J. , Defer C. , Mercier B. , Férec C. , Loiseau P. , Portelette E. , Mariotti M. , Lerable J. , Rouger P. , Pawlotsky J.‐M.
Published: March 2003

BACKGROUND: The systematic screening for several blood‐borne viral genomes in blood donations is a complementary safety measure planned or discussed by the national authorities of several countries.

STUDY DESIGN AND METHODS: To appreciate the feasibility of such screening using pooled samples, a multicenter study of real‐time simulation has been performed on the model of hepatitis C virus (HCV) infection. Four blood transfusion center laboratories and two research and diagnosis laboratories simultaneously screened, by several HCV RNA polymerase chain reaction assays, a panel of plasma sample pools of different sizes (500, 100, and 10 samples), collected from HCV‐infected or HCV‐ uninfected blood donors. One viremic plasma was introduced in each pool. HCV RNA was detected by in‐house polymerase chain reaction procedures or by standardized manual or semi‐automated polymerase chain reaction assays.

RESULTS: The results indicate the feasibility of sample pooling, which renders the screening for viral genomes by molecular biology techniques applicable in the short term and the importance of the experience of laboratory personnel in the use of molecular biology tools.

CONCLUSION: The improvement of standardized assays needs to be continued, and training of laboratory staff members appears to be a crucial step before systematic screening of blood donations for viral genomes by molecular biology techniques can occur.
Effect of filtration and storage of platelet concentrates on the production of the chemotaxins C5a, interleukin 8, tumor necrosis factor alpha, and leukotriene B4
Authors: Hetland G , Mollnes T.E , Bergh K , Hogasen K , Bergerud U.E , Solheim B.G
Published: March 2003

BACKGROUND: The production in platelet concentrates (PCs) of C3 activation products (C3bc), terminal complement complex (TCC), and chemotaxins C5a, interleukin (IL)‐8, tumor necrosis factor alpha (TNFalpha), and leukotriene B4 (LTB4) and the proposed reduction in concentration of the chemotaxins by white cell reduction were examined. STUDY DESIGN AND METHODS: Samples were collected from supernatants of PCs produced by apheresis (apheresis PCs) or from buffy coats (BC PCs) immediately after the production, after white cell‐reduction filtration on Day 1, and after 5‐day storage, and examined by enzyme immunoassays. RESULTS: Complement was activated in all PCs during storage, and the concentration of activation products was not influenced by prestorage filtration. In prestorage white cell‐reduced BC PCs, only C3bc levels increased. Levels of IL‐8, TNFalpha, and LTB4 increased during storage of apheresis PCs, but not in filtered units, except for LTB4. In contrast, levels of IL‐8 decreased after storage of filtered BC PCs. C5a correlated significantly with IL‐8, which also correlated with TNFalpha and LTB4. CONCLUSION: Both C5a and TNFalpha generation in apheresis PCs seem to induce white cell IL‐8 production, which mediates cellular LTB4 release. Prestorage white cell reduction is recommended for reducing chemotactic cytokine and leukotriene levels in all PCs. Production of BC PCs is recommended to achieve less complement activation, which is not affected by filtration.
ISO 9000 case study: the role of the external consultant
Authors: Nevalainen D.E.
Published: March 2003

BACKGROUND: Increasingly, blood centers worldwide are turning toward the ISO (International Organization for Standardization) 9000 Quality System Standards to meet regulatory demands of public authorities, competitive demands within their industry, and cost requirements from their customers. CASE REPORT: The South Texas Blood and Tissue Center is the first blood center in North America to achieve registration to the ISO 9002 Quality System Standards. The role of the external consultant in initiating the process and as a facilitator during the implementation process is examined. CONCLUSION: To ensure the long‐term success and viability of any quality system, it is necessary for an organization to develop and implement its own program. External consultants can assist in many ways, but it is important that their role be a supporting one rather than one of direct implementation.
Severe hemolytic anemia due to multiple red cell alloantibodies after an ABO‐incompatible allogeneic bone marrow transplant
Authors: Lopez A. , de la Rubia J. , Arriaga F. , Jimenez C. , Sanz G.F. , Carpio N. , Marty M.L.
Published: March 2003

BACKGROUND: A patient who received an ABO‐incompatible allogeneic bone marrow transplant experienced three episodes of immune hemolytic anemia due to multiple red cell (RBC) alloantibodies. CASE REPORT: A 41‐year‐ old man with chronic myeloid leukemia received an ABO‐incompatible bone marrow graft from his HLA‐identical brother. Selective removal of RBCs from donor marrow before transfusion was performed by centrifugation using a continuous‐flow blood cell separator. The patient was given group O Rh‐positive RBCs and group A Rh‐positive platelets. Prophylaxis for graft‐versus‐host disease consisted of cyclosporine and methotrexate. The patient experienced three hemolytic episodes, on Days 21, 35, and 160 which were due to different RBC alloantibodies (anti‐K, anti‐Jk(b), anti‐M, IgG anti‐A) produced by host lymphocytes surviving the conditioning regimen. RESULTS: The patient was group O, Jk(b‐), and the marrow donor was group A, Jk(b+). After the first hemolytic episode (Day 21), immunohematologic studies showed group O RBCs and a positive direct antiglobulin test (IgG+, C3d+). Antibody screening test and eluate studies detected anti‐M, anti‐Jk(b), and anti‐K. During the second hemolytic episode (Day 35), the patient's blood group showed a mixed population of group A and group O RBCs. The direct antiglobulin test was positive (IgG+, C3d+). Anti‐M, anti‐Jk(b), and IgG anti‐A were detected in the serum. Eluates made from the recipient's RBCs showed the same specificity as serum antibodies. During the third hemolytic episode (Day 160), a mixture of group O and group A RBCs was still present, the direct antiglobulin test was positive (IgG+, C3d‐), and anti‐Jk(b) and IgG anti‐A were observed in the serum and in an eluate made from the patient's RBCs. CONCLUSION: This is the first reported case of severe immune hemolytic anemia due to multiple RBC alloantibodies after an allogeneic bone marrow transplant. The time of appearance and the specificity of the antibodies strongly suggest that they were produced by residual recipient lymphoid cells.
Platelet collection with the Amicus and the AS.TEC 204 blood cell separators
Authors: Moog R. , Muller N. , Goergens D.
Published: March 2003

BACKGROUND: Platelet collection with two new blood cell separators was compared with respect to platelet yield, separation efficiency, white cell (WBC) contamination, and measures of component quality. Both machines offer a menu for the collection of a desired platelet yield. The harvesting of a definite platelet yield is a means of collecting a standardized therapeutic platelet dose. STUDY DESIGN AND METHODS: One hundred seventy‐two platelet collections were carried out with different protocols in the two devices (Amicus and AS.TEC 204). For that reason, healthy donors underwent plateletpheresis procedures using the continuous‐ and discontinuous‐flow systems. Separations with a definite endpoint volume and the option of collecting a desired platelet yield of 5.5 × 10(11) in the Amicus and 3.3 × 10(11) in the AS.TEC 204 machine were investigated. RESULTS: With the Amicus blood cell separator, platelet yield (5.02+/−0.46 × 10[11]) and separation efficiency (66.9+/−6.2%) were significantly higher than those with the AS.TEC 204 (3.02+/−0.62 × 10[11] and 46.4+/−6.7%, respectively). There was less deviation between the platelet yield and the desired platelet yield in the AS.TEC 204 machine than in the Amicus device. WBC contamination was 2.06+/−2.91 × 10(6) in components obtained from the AS.TEC 204 machine, and 87.5 percent of the platelet concentrates had a WBC content <5 × 10(6). None of the components from the Amicus device contained more than 5 × 10(6) WBCs. Except for platelet morphology score, measures of component quality were almost equal in the platelet concentrates obtained from the studied machines. CONCLUSION: Platelet yield and separation efficiency were significantly higher in the Amicus blood cell separator. The collection of a desired platelet yield was better achieved with the AS.TEC 204 device.
Hemagglutination inhibition of Cromer blood group antibodies with soluble recombinant decay‐accelerating factor
Authors: Daniels G. L. , Green C. A. , Powell R. M. , Ward T.
Published: March 2003

BACKGROUND: Cromer blood group antigens are located on decay‐ accelerating factor (DAF, CD55), which contains four short consensus repeats (SCRs). Cromer system antibodies may be of clinical significance in blood transfusion.

STUDY DESIGN AND METHODS: Soluble recombinant DAF (srDAF) constructs, consisting of all four SCRs or of only two SCRs, were expressed in the yeast Pichia pastoris. They are used in hemagglutination‐inhibition tests with Cromer system antibodies and with DAF‐specific monoclonal antibodies.

RESULTS: The srDAF inhibited hemagglutination by all Cromer system alloantibodies in undiluted serum. Antibodies to antigens of other blood group systems were not inhibited by the srDAF. Hemagglutination‐inhibition tests with domain‐deleted srDAF showed that UMC is on SCR‐4 and confirmed that Tca, TcaTcb, and WESb are on SCR‐1; Dra is on SCR‐3; and Cra is on SCR‐ 4.

CONCLUSIONS: Hemagglutination inhibition with srDAF is useful in the recognition of antibodies that belong to the Cromer blood group system and facilitates pretransfusion testing. This use of domain‐deleted srDAF provides an easy method of determining epitope location on DAF and is an aid to more precise identification of Cromer system antibodies.
An Israeli family with six cisAB members: serologic and enzymatic studies
Authors: Bennett M. , Levene C. , Greenwell P.
Published: March 2003

BACKGROUND: The cisAB blood type is a rare phenomenon in which both the A and B blood types are inherited from a single parent. Several forms of this phenotype have been characterized that differ with respect to serologic reactions and the activities of the gene‐encoded blood group A and B transferases.

STUDY DESIGN AND METHODS: The cisAB blood type was suspected when a baby typed as blood group O was born to a mother whose blood group was AB. Family studies revealed four generations in whom the pattern of inheritance could be explained only by the inheritance of the cisAB genotype. Blood and saliva samples from the family were tested serologically and assayed for the relevant glycosyltransferases. Samples suitable for DNA analysis were not available.

RESULTS: Six family members were shown serologically to be of the cisAB type. The A and B transferases in the sera of these individuals were 20 to 35 and 25 to 50 percent of those obtained for A and B individuals, respectively. The enzymic characteristics of the A and B transferases were determined. The A transferase in the sera of the cisAB persons did not bind to agarose beads.

CONCLUSION: The family described carries the cisAB gene, which encodes production of A and B transferases that differ from those of “normal” A, AB, and B controls. This variant has properties that are distinctly different from those described in other reports.
Improved progenitor assay standardization using peripheral blood progenitor cells from a donor treated with granulocyte–colony‐stimulating factor
Authors: Burger S.R. , Kadidlo D. , McCullough J.
Published: March 2003

BACKGROUND: Progenitor assays are the principal method for evaluating hematopoietic cell function. The magnitude of assay variability and the assay steps contributing to variability were determined, and modifications intended to increase assay consistency were evaluated.

STUDY DESIGN AND METHODS: Assays were performed using a serum‐free progenitor assay medium with cells plated at 5.0 × 104 and 1.0 × 105 cells per plate. A peripheral blood progenitor cell component collected from a normal donor after administration of granulocyte–colony‐stimulating factor was divided into identical aliquots. Each experiment involved at least 5 technologists, each performing assays in duplicate on five aliquots, with each person scoring all assay plates. Three sample preparation methods were tested: 1) ficoll mononuclear cell enrichment and sample dilution, 2) sample dilution without ficoll separation, and 3) sample dilution without ficoll separation, with cell counts performed before and after each dilution step, dilution volumes calculated on the basis of each cell count, automated electronic pipettors used in dilution steps, and colony frequency calculated on the basis of cell counts from the final specimen.

RESULTS: Global variability for colony‐forming units–granulocyte‐macrophage, represented by the percentage of CV for all specimens and all technologists, was 89.6 percent at 5.0 × 104 cells per plate and 81.3 percent at 1.0 × 105, when ficoll separation was used. Subjective differences in scoring plates did not account for most of the variability observed, as results for any individual plate read by multiple technologists had a mean CV of 15.6 percent and 19.7 percent at the two plating concentrations. Method 3 resulted in the greatest improvement, reducing CV to 24.4 percent at 5.0 × 104 cells per plate and to 24.2 percent at 1.0 × 105 cells per plate. Similar results were obtained for erythroid–burst‐forming units.

CONCLUSIONS: Baseline assay results were extremely inconsistent. Interindividual differences in colony interpretation did not contribute significantly to assay variability, although sample preparation and plating did. Improved control over cell concentration decreased assay variability by 70 to 73 percent.
Effects of filtration and gamma radiation on the accumulation of RANTES and transforming growth factor‐β1 in apheresis platelet concentrates during storage
Authors: Fujihara M. , Ikebuchi K. , Wakamoto S. , Sekiguchi S.
Published: March 2003

BACKGROUND: Platelet‐derived biologic response modifiers (BRMs) including RANTES and transforming growth factor (TGF)‐β1 accumulate in platelet components during storage because of platelet activation, and they may play a causative role in nonhemolytic febrile transfusion reactions. The majority of PCs with high unit values are provided by single donor apheresis in Japan.

STUDY DESIGN AND METHODS: RANTES and TGF‐β1 levels in platelet units prepared from single‐donor apheresis platelet concentrates (apheresis PCs) and units from whole blood (buffy coat PCs) were investigated. The effects of prestorage and poststorage filtration and gamma radiation on the levels of RANTES and TGF β1 in the supernatant of apheresis PCs during storage were also examined.

RESULTS: The levels of RANTES and TGF‐β1 increased during storage from Day 0 to Day 5. The levels of RANTES and of TGF‐β1correlated with the platelet concentration (p<0.01), but not with the residual white cell concentration in apheresis PCs that were not white cell reduced by filtration (p>0.05). In addition, there was a correlation between RANTES and TGF‐β1 levels (p<0.01). In white cell‐reduced apheresis PCs using negatively charged filters as well as in gamma‐radiated apheresis PCs, the levels of these two BRMs did not differ at any storage time from those of untreated apheresis PCs. Filtration of apheresis PCs with negatively charged filters after 3 days of storage significantly (p<0.05) reduced the levels of RANTES, but not of TGF‐β1. There was no reduction in the levels of RANTES and TGF‐β1 levels by positively charged filters. The RANTES levels in buffy coat PCs were slightly higher than but not significantly different from those of apheresis PCs during storage, except for the level on Day 1. There were no differences in the TGF‐β1 levels in apheresis and buffy coat PCs during storage.

CONCLUSION: Prestorage filtration and gamma radiation had neither preventive effects on the accumulation of RANTES and TGF‐β1 nor adverse effects on platelet activation. Negatively charged filters might be useful for the reducing the levels of RANTES in stored apheresis PCs.
A comparison of the Aplysia lectin anti‐I specificity with human anti‐I and several other I‐detecting lectins
Authors: Gilboa‐Garber N. , Sudakevitz D. , Levene C.
Published: March 2003

BACKGROUND: Lectins displaying blood group specificity are important for blood group typing and antigen recognition. Their use in blood banks is especially widespread in situations where there is a shortage of specific antisera. This report describes the efficiency of Aplysia gonad lectin as a reliable reagent for the detection of I antigen, which is common on adult human cells but reduced in fetal, newborn, and rare adult red cells.

STUDY DESIGN AND METHODS: The selective hemagglutinating activity of the Aplysia lectin was compared with that of human anti‐I and several I‐reactive lectins, including two plant lectins, one galactophilic microbial lectin, and bovine spleen galectin.

RESULTS: The comparison has revealed that Aplysia gonad lectin, like human anti‐I, strongly agglutinates and adsorbs to adult I‐positive red cells, differentiating between them and fetal or rare I‐negative adult red cells (although with less of a difference). In contrast to the plant and microbial lectins examined, its I‐affinity does not depend on the presence of ABH or P system antigens and it clearly detects higher I antigen expression in Oh red cells. The hemagglutinating activity of Aplysia lectin as that of all the I‐detecting proteins is enhanced at 4°C, but unlike the human anti‐I Aplysia lectin‐induced hemagglutination is stable at room temperature.

CONCLUSIONS: The Aplysia lectin is a reliable anti‐I reagent, which strongly agglutinates I‐positive adult human red cells irrespective of their ABH or P system antigens. This lectin is usable at room temperature.