bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative analysis of six different white cell‐reduction filters for packed red cells
Authors: Bontadini A. , Fruet F. , Tazzari P.L. , Lollini P.‐L. , Conte R.
Published: March 2003

Background: The reduction of white cells in blood components before transfusion by filters with at least 3 log10 depletion may prevent adverse transfusion reactions such as HLA alloimmunization, febrile reactions, transmitted infections, and immunomodulation. A new generation of filters with 4 log10 depletion is now available.

Study Design and Methods: The aim of this study is to compare the efficiency of white cell reduction by six commercial filters for packed red cells with 3 and 4 log10 depletion (claimed by manufacturers). The analysis of white cell concentration in the white cell‐reduced units was performed by flow cytometry and with a Nageotte chamber.

Results: The last generation of filters (BPF4, RC400, R01 Plus) show mean residual white cell numbers of 0.18 ± 0.14, 0.26 ± 0.21, and 0.25 ± 0.15 × 106, respectively, by flow cytometric analysis and 0.05 ± 0.04, 0.18 ± 0.15, and 0.38 ± 0.23 × 106, respectively, by Nageotte chamber evaluation. The 3 log10 depletion filters (R01, Leucostop‐4LT‐ mono, R200) have mean residual white cell numbers of 1.41 ± 0.92, 2.4 ± 1.99, and 1.05 ± 0.64 × 106, respectively, by flow cytometric analysis and 3.56 ± 1.7, 1.67 ± 1.3, and 3.21 ± 4.1 × 106, respectively, by Nageotte chamber evaluation. The data show that the BPF4, RC400, and R01 Plus filters are likely to be more efficient by 1 log10 reduction than the R01, Leucostop‐4LT‐mono, and R200 filters.

Conclusion: The most recent generation of filters is able to deplete white cells from packed red cells by 4 log10; in particular, with one of the filters, the residual WBC content was less than 0.5 × 106 per unit in all experiments, while two other filters reached that level in 9 of 10 experiments.
Blood group A immunodeterminants on human red cells differ in biologic activity and sensitivity to alpha‐N‐acetylgalactosaminidase
Authors: Hoskins L.C. , Larson G. , Naff G.B.
Published: March 2003

BACKGROUND: Epitopes of blood group A antigen can be enzymatically cleaved from red cells (RBCs), but the extent of cleavage required for normal survival in allogeneic blood transfusion recipients is unknown. Therefore, the cleavage rates were studied for A antigen epitope binding of 1) complement‐activating anti‐A, 2) Dolichos biflorus anti‐ A, lectin, and 3) hemagglutinating anti‐A during incubation with a purified alpha‐N‐acetylgalactosaminidase, E.C. 3.2.1.49 (alpha‐ GalNAc'ase).

STUDY DESIGN AND METHODS: Suspensions of group A RBCs were incubated with alpha‐GalNAc'ase. Cells were removed at intervals, washed, and tested for loss of binding by monoclonal, polyclonal, and complement‐activating anti‐A, D. biflorus anti‐A1 lectin, and Ulex europaeus anti‐H lectin.

RESULTS: A epitopes binding D. biflorus lectin were highly susceptible to alpha‐GalNAc'ase; simultaneously with their loss, binding with U. europaeus lectin emerged. Loss of complement‐ mediated hemolysis was slower. A epitopes binding hemagglutinating anti‐ A were most resistant. Cleavage of A epitopes from membrane glycosphingolipids with short oligosaccharide chains was similarly resistant. Rates of cleavage from A1 and A2 RBCs were similar.

CONCLUSION: RBC epitopes of blood group A differ in susceptibility to cleavage and biologic reactivity, which suggests that subsets mediating important biologic functions exist on functionally and topographically distinct membrane glycoconjugates.
In vitro and in vivo effects of prestorage filtration of apheresis platelets
Authors: Sweeney J. D. , Holme S. , Stromberg R. R. , Heaton W. A. L.
Published: March 2003

BACKGROUND: The effect of prestorage filtration on the quality of apheresis platelet concentrates stored for transfusion is undetermined. STUDY DESIGN AND METHODS: Investigation of 11 plateletpheresis components used a concurrent paired‐study design. On the day of collection, each component was equally divided into two suspensions; one half was filtered, and the other half was not. Each suspension was stored for 5 days. In vitro testing was performed on the day of collection (Day 0) for cell counts and on Day 5 for measurements of lactate, glucose, blood gases, pH, platelet ATP, hypotonic stress ratio, extent of shape change in response to ADP, tissue necrosis factor alpha, interleukin 8, interleukin 1 alpha, interleukin 1 beta, interleukin 6, and platelet surface glycoproteins by flow cytometry. At the end of the 5‐day period, a sample was taken from each of the two suspensions, radiolabeled with either 51Cr or 111In, and transfused concurrently. Posttransfusion samples were drawn for measurements of recovery and platelet survival and for functional assessment of the ex vivo ability of the circulating radiolabeled platelets to aggregate in response to ADP. RESULTS: The apheresis component had a mean platelet yield of 3.2 +/− 0.4 × 10(11) and a white cell yield ranging from 1 × 10(5) to 1 × 10(8), with a median of 2 × 10(7). Filtration resulted in a platelet loss of approximately 10 percent and a variable 2 to 3 log10 reduction in white cell content. No significant differences between filtered and unfiltered suspensions in paired t tests that would likely have an impact on platelet quality were observed in the in vitro tests. The in vivo recovery and survival were highly similar and not statistically different in filtered and unfiltered paired suspensions: the mean difference was 1.2 +/− 4.0 percent for recovery and 7.0 +/− 15 hours for survival. The functional assessment by aggregation to ADP showed no difference between filtered and unfiltered suspensions. A small decrease in tumor necrosis factor alpha and interleukin 8 was evident in the filtered suspension as compared to levels in the unfiltered suspensions. CONCLUSION: Prestorage white cell reduction in apheresis components resulted in WBC reduction by several log10 with no evident adverse effect on platelet viability or function.
White cell‐reduced platelet concentrates prepared by in‐line filtration of platelet‐rich plasma
Authors: Sweeney J. D. , Holme S. , Heaton W. A. L. , Nelson E.
Published: March 2003

BACKGROUND: The importance of white cell (WBC) reduction in platelet concentrates (PCs) for component quality is undetermined. STUDY DESIGN AND METHODS: Eleven paired components, each derived from one of two whole‐blood units given by a single donor on the same day, were studied. One PC was WBC reduced by filtration with an in‐line, integral, prototype filter, and the other was produced from unfiltered platelet‐rich plasma (PRP) by a standard method. In vitro tests performed on Day 1 and Day 5 were blood gases, plasma lactate, glucose, platelet ATP, mean platelet volume, morphology score, hypotonic stress ratio, extent of shape change in response to ADP, and beta‐ thromboglobulin. After 5 days of storage, each component pair was labeled with 51Cr or 111In and transfused for the estimation of percent recovery and survival. RESULTS: PCs using the in‐line, prototypic filter had a platelet loss of approximately 15 percent and a variable 1 to 3 log10 reduction (average, 95%) in WBC content. The variation in filter WBC removal was related to PRP WBC content and indicated that the filter did not have the capacity for a 3 to 4 log10 removal when PRP WBC content exceeded 1 × 10(8). The in vitro and in vivo measures of platelet quality showed no meaningful differences between filtered and unfiltered PCs by paired t test. The mean differences in posttransfusion percent recoveries and survivals were 0.9 +/− 2.9 percent and 4 +/− 13 hours, respectively. Additional studies were performed using a larger filter with improved capacity. Those studies (n = 18) showed a significant improvement in filtration time and platelet yield and a consistent 3 to 4 log10 reduction in WBCs. Filtration time was 6.6 +/− 2.7 minutes, total PC WBCs were 9.6 +/− 4.6 × 10(4), and total PC platelets were 7.8 +/− 1.8 × 10(10) (mean +/− SD). CONCLUSION: Prestorage filtration of PRP and the preparation of filtered platelets do not result in any significant beneficial or adverse effect on subsequent platelet quality. With the large‐capacity filter, consistent WBC reduction and good platelet yields are achieved.
Long‐term follow‐up of blood donors with indeterminate human immunodeficiency virus type 1 results on Western blot
Authors: Jackson J. B. , Hanson M. R. , JOhnson G. M. , Spahlinger T. G. , Polesky H. F. , Bowman R. J.
Published: March 2003

BACKGROUND: At present, tens of thousands of United States blood donors who are at low risk for human immunodeficiency virus type 1 (HIV‐1) infection are indefinitely deferred. These persons are repeatably reactive for HIV‐1 antibody in enzyme immunoassay (EIA) and are indeterminate in Western blot. STUDY DESIGN AND METHODS: To determine the significance and persistence of anti‐HIV‐1 reactivity in plasma from volunteer blood donors with HIV‐1‐indeterminate Western blots, 66 donors were retested for HIV‐1 antibody by the same manufacturers' EIA and Western blot 5 to 7 years after the initial Western blot. In addition, donors' peripheral blood mononuclear cells were tested by polymerase chain reaction (PCR) for HIV‐1 DNA gag sequences. RESULTS: Thirty‐five (53%) of 66 donors were still repeatedly reactive for HIV‐1 on EIA and indeterminate on Western blot, 23 (35%) were negative on EIA and indeterminate on Western blot, 7 (11%) were negative in EIA and Western blot, and 1 (2%) was repeatedly reactive on EIA and negative on Western blot. Donors with persistently indeterminate Western blots had a band pattern nearly identical to that on the original Western blot. No donor was positive in Western blot, p24 antigen, or PCR testing. No donor had signs or symptoms of HIV‐1 infection. CONCLUSION: Long‐term follow‐up of Western blot‐indeterminate blood donors does not reveal evidence of HIV‐infection. A mechanism to return these donors to the donor pool should be considered.
Comparison of the efficacy of blood and polyethylene glycol‐hemoglobin in recovery of newborn piglets from hemorrhagic hypotension: effect on blood pressure, cortical oxygen, and extracellular dopamine in the brain
Authors: Song D. , Olano M. , Wilson D. F. , Pastuszko A. , Tammela O. , Nho K. , Shorr R.G.L.
Published: March 2003

BACKGROUND: Successful blood substitutes, when infused in place of an equal volume of whole blood, provide similar delivery of oxygen to the tissues without introducing abnormalities in cellular metabolism.

STUDY DESIGN AND METHODS: Equal volumes of whole blood (control), polyethylene glycol‐hemoglobin solution at 6 g per dL, dextran solution, and physiologic saline were compared for their ability to reverse the effects of hemorrhagic hypotension on oxygenation and dopamine metabolism in the brain of newborn piglets. The decrease in mean arterial blood pressure was used as a measure of the hemorrhagic insult. Cerebral oxygen pressure was determined optically by the oxygen‐ dependent quenching of phosphorescence, and the extracellular level of dopamine in the corpus striatum was determined by in vivo microdialysis.

RESULTS: Following a 2‐hour stabilization after implantation of the microdialysis probe in the corpus striatum, the mean arterial blood pressure was decreased from 88 +/− 7 torr (control) to 42 +/− 5 torr by the removal of blood in a stepwise manner, over a period of 60 minutes. Decrease in mean arterial blood pressure caused a progressive stepwise decrease in cortical oxygen pressure from 48 +/− 5 torr to 16 +/− 4 torr at the end of bleeding. As a consequence of the decrease in oxygen pressure, extracellular dopamine increased progressively to about 2300 percent of the control value. When a volume of blood equal to that removed was returned and bicarbonate was injected to help correct arterial pH, blood pressure, cortical oxygen pressure, and extracellular dopamine all returned within the 20‐ to 30‐ minute recovery period to values not significantly different from control values. An equal volume of polyethylene glycol‐hemoglobin solution, even with significantly lower hemoglobin content than whole blood, gave results comparable to those with whole blood.

CONCLUSION: Polyethylene glycol‐hemoglobin solution, like whole blood but in contrast to physiologic saline or dextran solution, was capable of returning the mean arterial blood pressure, cortical oxygen pressures, and extracellular dopamine nearly to control levels after acute blood loss in newborn piglets.
Analysis of the priming activity of lipids generated during routine storage of platelet concentrates
Authors: Silliman C. C. , Dickey W. O. , Paterson A. J. , Thurman G. W. , Clay K. L. , Johnson C. A. , Ambruso D. R.
Published: March 2003

BACKGROUND: Compounds generated during the routine storage of platelet concentrates may have deleterious effects on the transfusion recipient.

STUDY DESIGN and METHODS: Daily plasma samples from platelet concentrates, both apheresis platelets and those separated from whole blood, were obtained serially during routine storage. These plasma samples were assayed for their ability to prime the NADPH oxidase in isolated human neutrophils. Quantitative and qualitative analysis of the priming agents was completed by lipid extraction, high‐pressure liquid chromatography separation, and gas chromatography/mass spectroscopy.

RESULTS: Compounds were generated in both apheresis and whole‐blood platelets that significantly primed the NADPH oxidase after 24 and 48 hours of storage, respectively. The priming activity was maximal by component outdate: 2.6‐fold that of the buffer‐treated control neutrophils (apheresis) and 3.9‐fold that of the buffer‐treated control neutrophils (whole blood). These agents were generated by cellular constituents, as stored plasma did not demonstrate such priming activity. Inhibition of this priming activity by WEB 2170, a specific platelet‐activating factor receptor antagonist, suggested that the observed priming involved the platelet‐activating factor receptor. A portion of the priming activity from platelet concentrates was organically extractable: 69 percent of that from apheresis platelets and 46 percent of that from whole‐blood platelets. Further purification of the lipid's priming activity by normal‐phase high‐pressure liquid chromatography demonstrated a single peak of priming activity at the retention time of lysophosphatidylcholines. Because 46 percent of the priming activity from whole‐blood platelets was chloroform insoluble and because it has been reported that interleukin 8 is generated during routine storage of whole‐blood platelets, the effects of interleukin 8 on the NADPH oxidase were examined. Recombinant monocyte interleukin 8 rapidly primed the oxidase but was not inhibited by WEB 2170.

CONCLUSION: Lipids were generated during the routine storage of platelet concentrates that prime the NADPH oxidase, and they may play a role in the severe complications of transfusion therapy. Other non‐ lipid compounds, such as interleukin 8, that are generated in whole‐ blood platelets may also contribute to the observed priming activity of plasma.
Unexplained hepatitis C virus antibody seroconversion in established blood donors
Authors: Atrah H.I. , Ala F.A. , Ahmed M.M. , Hutchinson F. , Cough D. , Baker K.
Published: March 2003

Background: Understanding of the epidemiology and natural history of hepatitis C virus (HCV) infection is incomplete without reference to the early phase of infection. The prevalence of HCV infection is well documented in numerous reports. The seroconversion pattern in previously antibody‐negative blood donors provides a model for the study of the incidence and transmission of HCV infection.

Study Design and Methods: Records of HCV antibody tests at the West Midlands Blood Transfusion Centre were reviewed to determine the seroconversion rate in 1994 among previously anti‐HCV‐negative blood donors. Seroconverting donors were counseled to investigate the possible routes of infection.

Results: In 1994, blood donations (n = 256,935) were collected from 149,370 donors; 24 donors (0.016%; 1/6224) were positive in the screening enzyme‐linked immunosorbent assay (ELISA) and the third‐ generation recombinant immunoblot assay (RIBA‐3). Two donors previously negative for HCV antibody in ELISA were positive in both tests in 1994. Four donors positive in ELISA and indeterminate in RIBA‐3 in 1993 reacted positively in both tests in 1994. One donor negative for HCV antibody on previous screening reacted positively in ELISA and was indeterminate in RIBA‐3 in 1994 and has become positive in both tests in 1995. A further 43 donors negative for HCV antibody on previous screening reacted positively in ELISA and were indeterminate in RIBA‐3 in 1994.

Conclusion: Documented seroconversion can take place in the absence of exposure to recognizable risk factors for the infection. The index donation or the donation immediately preceding seroconversion may be positive for HCV RNA in the polymerase chain reaction.
Infection by hepatitis C virus through contaminated intravenous immune globulin: results of a prospective national inquiry in France
Authors: Lefrere J.J. , Loiseau P. , Martinot‐Peignoux M. , Mariotti M. , Ravera N. , Thauvin M. , Marcellin P. , Janot C.
Published: March 2003

BACKGROUND: A recent hepatitis C virus (HCV) outbreak has been suspected of being caused by an infusion of intravenous immune globulin.

STUDY DESIGN AND METHODS: Three laboratories were mandated by the French regulatory agency to prospectively screen on a national scale those persons having received suspected batches: 233 exposed patients were recalled and tested for HCV antibody and for HCV RNA.

RESULTS: Nineteen patients (8.1%) were found positive for HCV RNA; 7 of these 19 were positive for the HCV antibody.

CONCLUSION: The link between HCV infection and intravenous immune globulin was reinforced by the overrepresentation of the 2b genotype (58%), which contrasts with the low prevalence of this genotype in France (1%).
A standardized method for calculating blood loss
Authors: Brecher M. E. , Monk T. , Goodnough L. T.
Published: March 2003

BACKGROUND: The estimation of blood loss for a surgical procedure is both poorly reproducible and typically underestimated. Therefore, comparison of surgical transfusion outcomes such as blood loss and amount of blood transfused from one institution to another, or from one surgeon to another, is difficult. Recently, mathematical modeling has contributed to our understanding of transfusion strategies. STUDY DESIGN AND METHODS: A mathematical model of blood loss for a surgical hospitalization was developed on the basis of recently described mathematical principles for blood loss and hemodilution. The model was designed so that the calculation of blood loss would be based on easily measured factors such as the patient's blood volume, the number and type of red cell units transfused, the initial hematocrit, the discharge hematocrit, the transfusion trigger, the volume of intraoperatively salvaged blood transfused, and the amount of hemodilution performed. The calculated blood loss was then compared with the intraoperative blood loss actually estimated by the anesthesiologist in 250 consecutive patients who underwent radical retropubic prostatectomy. RESULTS: The mathematical equations were placed in a computer model to allow rapid calculation of a particular patient's blood loss. Figures were derived from the computer modelling to facilitate rapid manual calculation of the blood loss. There was a significant relation (p < 0.001) between the calculated blood loss for the hospitalization and the estimated intraoperative blood loss. However, the calculated blood loss was on average 2.1 times the intraoperative blood loss estimated by the anesthesiologist. CONCLUSION: The use of such mathematical modeling to rapidly estimate a patient's blood loss has the potential to allow ready, objective comparisons between sites and even surgeons. It also allows for a more judicial and informed decision as to what (if any) blood should be available or what blood‐conservation techniques should be employed for a specific patient.
IgA red cell autoantibodies and autoimmune hemolysis
Authors: Sokol R.J. , Booker D.J. , Stamps R. , Booth J.R. , Hook V.
Published: March 2003

BACKGROUND: The objective of the study was to examine the interrelationships and clinical significance of IgA red cell antibodies in the autoimmune response. STUDY DESIGN AND METHODS: The records of 5235 patients referred to an immunohematology center over a 14‐year period were critically examined for patients who had IgA autoantibodies, defined as elutable IgA immunoglobulins that would rebind to normal cells. RESULTS: One hundred twenty‐four patients (61 male) aged 6 to 98 years had warm‐reacting IgA autoantibodies. In 75 individuals, these were idiopathic; neoplasms were the most common associated conditions in the individuals with secondary IgA autoantibodies. IgA was the only immunoglobulin present in 6 patients; all others also had IgG and/or IgM coating their cells, and 102 individuals also had increased amounts of cell‐bound complement. In a comparison by chi‐square test of populations with haptoglobins of < 0.1 g per L, IgA was shown to act synergistically with IgG in producing hemolysis (p < 0.01). CONCLUSION: Autoimmune hemolysis due to IgA antibodies alone in rare, with red cell destruction occurring through mechanisms similar to those for IgG. Most commonly, IgA acts synergistically with other immunoglobulins (usually IgG) and complement; the hemolysis may be severe. Whether IgA autoantibodies alone can activate complement remains controversial, but increasing evidence suggests that they can, possibly via the alternative pathway, and that this activation may result in intravascular hemolysis.
Psoralen‐mediated photodecontamination of platelet concentrates: inactivation of cell‐free and cell‐associated forms of human immunodeficiency virus and assessment of platelet function in vivo
Authors: Margolis‐Nunno H. , Bardossy L. , Robinson R. , Ben‐Hur E. , Horowitz B. , Blajchman M. A.
Published: March 2003

BACKGROUND: Treatment of platelet concentrates (PCs) with psoralens and broad‐band ultraviolet A (UVA) radiation is being examined for the elimination of pathogens that might be present in donated blood. Previous studies have demonstrated the inactivation of cell‐free viruses and the maintenance of platelet integrity with common in vitro assays. STUDY DESIGN AND METHODS: Human immunodeficiency virus (HIV) in three forms‐cell‐free, activity replicating, and latently infected cell lines‐was added to PCs and treated with 50‐microgram per mL of 4′‐ aminomethyl‐4,5′,8‐trimethylpsoralen (AMT), 0.35 mM rutin, and broad‐ and narrow‐band UVA light (320‐400 nm and 360–370 nm [UVA1], respectively). The inactivation of added HIV was assessed in tissue culture; platelet hemostatic activity was assessed in thrombocytopenic rabbits. RESULTS: Each form of HIV was inactivated completely (> or = 10(5) infectious units) on treatment with 30 J per cm2 of UVA1 light. Similar results were obtained on treatment of 2.5 mL of PCs in test tubes or intact PC units (50 mL) in blood bags. Latently infected cell lines were substantially more sensitive than cell‐free HIV or HIV that was actively replicating. Human platelets treated with 40 J per cm2 of UVA1 light had a fully corrected bleeding time shortly after treatment or after 5 days' storage, as assessed in thrombocytopenic rabbits. Platelet hemostatic function began to decrease with 81 J per cm2 of UVA1 light and was abolished with 113 J per cm2. At similar fluences, broad‐band UVA light was more injurious to platelets than was UVA1 light. CONCLUSION: HIV transmission might be eliminated by PCs after treatment with AMT and UVA1 light and without a reduction in platelet hemostatic function.
Frequency of immediate adverse effects associated with apheresis donation
Authors: McLeod B. C. , Price T. H. , Owen H. , Ciavarella D. , Sniecinski I. , Randels M. J. , Smith J. W.
Published: March 2003

BACKGROUND: Apheresis donation is considered safe, but the incidence of adverse effects has not been determined in a large multicenter series of donations with modern instruments.

STUDY DESIGN AND METHODS: The Hemapheresis Committee of the American Association of Blood Banks devised a uniform questionnaire that asked about 32 specific adverse effects. Transient paresthesia and mild vasovagal events were excluded. A survey was conducted in 1995; 17 centers returned 19,611 responses concerning 250 to 2,000 consecutive apheresis donations per center.

RESULTS: Six hundred adverse effects were reported in 428 donations (2.18% of donations). Pain or hematoma at a venipuncture site was the most common response (1.15% of donations); only 203 donations had other (nonvenipuncture) adverse effects (1.04%). Total and nonvenipuncture rates were, respectively, 4.84 and 2.92 percent for 2,295 first donations and 1.78 and 0.77 percent for 17,303 repeat donations (p < 0.001). Rates of nonvenipuncture symptoms in first and repeat donations were, respectively, citrate‐induced nausea and/or vomiting, 0.87 and 0.27 percent; tetany, 0.09 and 0.04 percent; pallor and/or diaphoresis, 1.87 and 0.32 percent; vasovagal nausea and/or vomiting, 0.87 and 0.13 percent; syncope and/or seizure, 0.39 and 0.04 percent; and chills and/or rigors, 0.31 and 0.01 percent. The overall rate of donor unconsciousness was 0.08 percent. Hemolysis was reported twice. Clotting or leakage occurred in 0.08 percent of donations, and inability to return blood occurred in 0.16 percent. No life‐threatening adverse effects were reported. Procedure‐specific nonvenipuncture rates were 1.05 percent of 17,584 platelet donations, 0.67 percent of 594 white cell donations, and 0.37 percent of 1,354 plasma donations. Center‐specific rates varied from 0.32 to 6.81 percent of donations for total adverse effects and from 0.11 to 2.92 percent of donations for nonvenipuncture events.

CONCLUSION: Apheresis donation is a safe undertaking, suitable for voluntary blood donors, with a very low risk of serious adverse effects. The risk of unconsciousness is lower than that found in many studies of whole‐blood donation.
Assessment of the clinical significance of anti‐Dob
Authors: Shirey R. S. , Boyd J. S. , King K. E. , Caturegli P. P. , Ness P. M. , Montgomery W. M.
Published: March 2003

BACKGROUND: Anti‐Dob is an uncommon antibody, and there are few data regarding its clinical importance. In the present case, the patient's transfusion management was based on both in vivo and in vitro assay results. CASE REPORT: A delayed hemolytic transfusion reaction was suspected in a 64‐year‐old white woman awaiting cardiac surgery when the transfusion of 1 unit of red cells failed to raise her hematocrit. Although direct antiglobulin tests were negative, antibody screening tests on samples drawn 9 days after transfusion were positive, and anti‐ Dob was identified, reacting to a titer of 4. 51Cr in vivo survival studies with incompatible Do(b+) red cells showed poor survival: 83.2 percent at 1 hour, 43 percent at 24 hours, and 29.6 percent at 48 hours and t1/2 = 19 hours (normal t1/2 = 25–35 days). A monocyte monolayer assay performed with the same incompatible Do(b+) donor red cells also indicated poor survival: 22 percent and 30 percent reactive monocytes, respectively, with and without the addition of complement (normal, 0‐ 3%). The patient was given 4 Do(b‐) red cell units without clinical signs or symptoms of a reaction. CONCLUSIONS: This example of anti‐Dob was implicated in a delayed hemolytic transfusion reaction. The 51Cr survival studies and monocyte monolayer assay results indicated that the anti‐Dob was clinically significant, requiring the use of Do(b‐) red cells for transfusion.
Identification and classification of the causes of events in transfusion medicine
Authors: Kaplan H. , Battles J.B. , Van der Schaaf T.W. , Shea C.E. , Mercer S. Q.
Published: March 2003

BACKGROUND: Transfusion medicine lacks a standard method for the systematic collection and analysis of event reports. Review of event reports from the Food and Drug Administration (FDA) showed a relative paucity of information on event causation. Thus, a causal analysis method was developed as part of a prototype Medical Event Reporting System for Transfusion Medicine (MERS‐TM). STUDY DESIGN AND METHODS: MERS‐TM functions within existing quality assurance systems and utilizes descriptive coding and causal classification schemes. The descriptive classification system, based upon current FDA coding, was modified to meet participant needs. The Eindhoven Classification Model (Medical Version) was adopted for causal classification and analysis. Inter‐rater reliability for the MERS‐TM and among participating organizations was performed with the development group in the United States and with a safety science research group in the Netherlands. The MERS‐TM was then tested with events reported by participants. RESULTS: Data from 503 event reports from two blood centers and two transfusion services are discussed. The data showed multiple causes for events and more latent causes than previously recognized. The distribution of causes was remarkably similar to that in an industrial setting outside of medicine that uses the same classification approach. There was a high degree of inter‐rater reliability when the same events were analyzed by quality assurance personnel in different participating organizations. These personnel found the method practical and useful for providing new insights into conditions producing undesired events. CONCLUSION: A generally applicable and reliable method for identifying and quantifying problems that exist throughout transfusion medicine will be a valuable addition to event reporting activity. By using a common taxonomy, participants can compare their experience with that of others. If proven as readily implementable and useful as shown in initial studies, MERS‐TM is a potential standard for transfusion medicine.
Mobilized CD34+ cells selected as autografts in patients with primary light‐chain amyloidosis: rationale and application
Authors: Comenzo RL , Michelle D , LeBlanc M , Wally J , Zhang Y , Kica G , Karandish S , Arkin CF , Wright DG , Skinner M , McMannis J
Published: March 2003

BACKGROUND: Concern about tumor cell contamination in stem cell preparations has led to the use of CD34+ cell selection as a means of purging. Increasing the number of CD34+ cells per leukapheresis may help to provide an adequate dose of CD34+ cells. STUDY DESIGN AND METHODS: The reverse transcriptase polymerase chain reaction (RT‐PCR) was employed to clone overexpressed clonotypic immunoglobulin light‐ chain variable region genes (Ig VL) from bone marrows of patients with primary light‐chain amyloidosis (AL). Patient‐specific primers were designed to evaluate stem cell collections for contamination. CD34+ cell selection was performed on components from AL patients who underwent mobilization with granulocyte‐colony‐stimulating factor (G‐ CSF) (filgrastim; 16 microg/kg/d for 4 days) and collection by large‐ volume leukapheresis (LVL;25L) on Days 4 and 5. The selected cells alone were transfused after patients received mephalan (200 mg/m2). RESULTS: Contamination was found in collections from 4 to 7 patients, which provided the rationale for a subsequent trial of CD34+ cell selection. The median number of CD34+ cells per kg collected on Days 4 and 5, and in toto, was 4.0 × 10(6)(1.1‐12.7), 7.9 × 10(6)(1.8‐12.7), and 10.7 × 10(6)(2.9‐25.4), respectively (n = 9 patients). The median yield per selection was 38 percent, with a purity of 85 percent (45‐ 97%), and the viability of CD34+ cells averaged 96.4 +/− 3.6 percent (n = 18 selections). The median number of CD34+ cells infused was 5.9 × 10(6) per kg (2.1‐10.1). In comparison with AL patients given unselected autografts, patients receiving selected CD34+ cells experienced similar reconstitution of neutrophils and platelets but slower lymphocyte recovery. CONCLUSION: Patients with AL often have contamination with clonotypic cells in their blood autografts. G‐CSF mobilization and LVL provide components that allow the selection of adequate doses of CD34+ cells. The use of CD34+ cells in patients with AL achieves rapid neutrophil and platelet recovery but delayed lymphocyte recovery. CD34+ cell selection is feasible in the treatment of AL, but its effectiveness in purging clonotypic cells remains to be ascertained.
Effects of hyperoxic ventilation on hemodilution‐induced changes in anesthetized dogs
Authors: Habler O.P. , Kleen M.S. , Hutter J.W. , Podtschaske A.H. , Tiede M. , Kemming G.I. , Welte M.V. , Corso C.O. , Batra S. , Keipert P.E. , Faithfull N.S. , Messmer K.F.
Published: March 2003

BACKGROUND: In subjects who have undergone acute preoperative normovolemic hemodilution (ANH), intraoperative hemorrhage is generally treated by immediate return of autologous blood collected during ANH. Simply increasing blood oxygen content by hyperoxic ventilation (HV, inspiratory fraction [FIO2] 1.0) might compensate for the acute anemia, allow further ANH, and delay onset of autologous blood return. STUDY DESIGN AND METHODS: This study 1) evaluated the effects of HV (FIO2 1.0) upon ANH to a hemoglobin (Hb) concentration of 7 g per dL in anesthetized dogs ventilated with room air and 2) compared the effects of subsequent profound ANH (Hb, 3 g/dL) with and without an intravenous perfluorocarbon emulsion (perflubron 60% wt/vol) versus those of autologous red cell transfusion. The results of the entire study are presented in two parts. Organ tissue oxygenation was assessed in skeletal muscle and liver, and systemic oxygenation status was evaluated. Myocardial contractility was deduced from left ventricular pressure‐volume relationship. Seven of 22 dogs underwent further hemodilution while breathing 100‐percent O2, for a determination of the Hb concentration at which HV‐induced effects were abolished. RESULTS: HV completely reversed the ANH‐induced increase in cardiac index (4.6 +/− 0.7 vs. 3.8 +/− 0.9 L/min/m2 before and during HV; p < 0.05) and partially reversed the decrease in systemic vascular resistance (1784 +/− 329 vs. 2087 +/− 524 dyn × cm‐5 × sec × m‐2; p < 0.05). Despite unchanged global O2 delivery, organ tissue oxygenation improved during HV (mixed venous partial pressure of O2: 40 +/− 3 vs. 59 +/− 7 torr; coronary venous pressure of O2: 30 +/− 4 vs. 43 +/− 6 torr; p < 0.05; liver surface: 31 +/− 11 vs. 39 +/− 13 torr; skeletal muscle surface: 30 +/− 14 vs. 41 +/− 22 torr; p < 0.05). This improvement was due to an increased contribution of physically dissolved O2 in plasma to O2 delivery (3.2 +/− 0.2% before HV vs. 14.6 +/− 1% during HV; p < 0.05) and O2 consumption (whole body: 6 +/− 1% vs. 47 +/− 8%, p < 0.05; myocardium: 4.3 +/− 0.9% vs. 31 +/− 6%, p < 0.05). The beneficial effects of HV were lost after an additional volume‐compensated exchange of 19 percent of blood volume (Hb, 5.6 g/dL). CONCLUSION: In anesthetized dogs ventilated with room air and hemodiluted to a Hb of 7 g per dL, simple oxygen therapy by HV (FIO2 1.0) rapidly improves tissue oxygenation and permits extended hemodilution to Hb of 5.8 g per dL until the HV‐induced effects are lost.
Lack of evidence of hepatitis C infection in 290 blood component recipients, demonstrated by several single‐antigen research immunoassays
Authors: Henrard D.R. , Berthillon P. , Scheffel J.W. , Ladaique P.L. , Moore B.S. , Pailhous M.C. , Finetti P.H. , Trepo C.
Published: March 2003

BACKGROUND: A group of 290 transfusion recipients enrolled in a prospective study of posttransfusion hepatitis was studied to determine the possibility of previously unrecognized hepatitis C virus (HCV) transmission. STUDY DESIGN AND METHODS: Before and after transfusion, blood specimens that were negative in first‐generation enzyme immunoassay (EIA) were tested by current commercial EIAs, several single‐antigen research EIAs, and supplemental tests. RESULTS: Current second‐ and third‐generation EIAs identified five subjects (1.7% of total) who had chronic hepatitis C before transfusion. Twenty additional sera had some reactivity with research EIAs. However, those results were the same before and after transfusion (n = 7), had reverted to partially reactive or nonreactive (n = 8), or could not be confirmed by serologic tests or polymerase chain reaction in follow‐up specimens (n = 5). CONCLUSIONS: Transient or restricted reactivity to HCV antigens measured by more sensitive research EIAs does not seem to correspond to recent HCV transmission by transfusion. Whether such reactivity could reflect remote HCV infection, with the potential for chronic or intermittent viremia, remains to be determined.
RHD genotyping in weak D phenotypes by multiple polymerase chain reactions
Authors: Legler T.J. , Maas J.H. , Blaschke V. , Malekan M. , Ohto H. , Lynen R. , Bustami N. , Schwartz D.W.M. , Mayr W.R. , Kohler M. , Panzer S.
Published: March 2003

BACKGROUND: Weak D phenotypes involve a quantitative variation of D. The genomic basis in weak D has been disputed, however.

STUDY DESIGN AND METHODS: Five sequence‐specific polymerase chain reactions (SSP‐ PCRs) on exons 2, 5, and 7 of the RHD gene were evaluated in 248 white and 98 Japanese blood donors and compared with the results obtained by amplification of intron 4 and serology. All methods and SSP‐PCR testing on the 3′ non‐coding region of the RHD gene were applied to the genotyping of 94 DNA samples derived from individuals expressing weak D phenotypes.

RESULTS: Concordant results were obtained with all genotyping and phenotyping methods in testing 201 D‐positive and 145 D‐ negative donors. Four of 94 weak D samples were typed as D‐negative by amplification of intron 4 and SSP‐PCR on exon 5. Phenotyping with monoclonal antibodies revealed a DVI category in one of these cases and DFR phenotype in three of these cases. One weak D sample, which reacted like normal D‐positive cells with all applied monoclonal antibodies, was typed falsely negative by SSP‐PCR on exon 5 because of a point mutation at nucleotide 667 (T–>G) that resulted in a Phe223Val amino acid substitution. In this individual, heterozygosity was found at two other amino acid positions (Glu233Gln and Val238Met) by restriction fragment length polymorphism analysis.

CONCLUSION: Genetic diversity in weak D phenotypes is rare. Only 1 of 90 true weak D phenotypes (1.1%) had a genetic variation in testing on seven gene regions of the RHD gene.
A recombinant peptide antigen line immunoassay optimized for the confirmation of Chagas' disease
Authors: Oelemann W.M.R. , Vanderborght B.O.M. , Verissimo Da Costa G.C. , Teixeira M.G.M. , Borges‐Pereira J. , De Castro J.A.F. , Coura J.R. , Stoops E. , Hulstaert F. , Zrein M. , Peralta J.M.
Published: March 2003

BACKGROUND: The transfusion of contaminated blood has become the major route of transmission for Chagas' disease in Brazil. Current screening tests are insensitive and yield conflicting results, while confirmatory assays do not exist. A line immunoassay (INNO‐LIA Chagas Ab [INNO‐LIA]) combining relevant, immunodominant recombinant and synthetic antigens on a single nylon membrane strip was evaluated for the serologic confirmation of Chagas' disease.

STUDY DESIGN AND METHODS: Sera from 1062 patients and healthy residents of four Brazilian regions endemic for Chagas' disease were used for test optimization. The established confirmation algorithm was evaluated with an independent set of positive (n = 75) and negative (n = 148) samples.

RESULTS: In the optimization phase, without an established comparative gold standard, the results with the INNO‐LIA were compared with those obtained in four other screening assays. In the validation phase, the INNO‐LIA showed a sensitivity of 100 percent (95% CI, 95.21‐100) and a specificity of 99.32 percent (95% CI, 96.29‐99.98) for well‐characterized sera. Moreover, its specificity reached 100 percent with a set of 40 sera obtained from patients with documented leishmaniasis. The interpretation criteria defined in this study indicated that the INNO‐LIA accurately detected the presence of antibodies to various specific antigens of Trypanosoma cruzi.

CONCLUSION: The INNO‐LIA Chagas Ab assay may become the first commercial assay to reliably confirm the presence of antibodies to T. cruzi.
Inhibition of a secondary human alloimmune response via the soluble active component of CD154 (CD40L) in severe combined immune‐deficient mice engrafted with human lymphocytes
Authors: Lazarus A.H. , Crow A.R , Freedman J. , Blanchette V. , Hannach B.
Published: March 2003

BACKGROUND: Alloimmunization requires a process known as co‐stimulation. An important co‐stimulatory pathway for most immune responses is mediated by the interaction of CD40 on antigen‐presenting cells with CD154 (CD40L) on host T cells. Blockade of this co‐stimulatory pathway simultaneous with exposure to challenge with HLA‐incompatible cells is hypothesized to inhibit alloimmunization.

STUDY DESIGN AND METHODS: Severe combined immune‐deficient (SCID) mice were reconstituted with human peripheral blood lymphocytes (Hu‐PBL‐SCID mice) from a subject primed to HLA antigens and challenged with HLA‐incompatible lymphocytes. Mice were challenged in the presence or absence of an 18‐kDa soluble recombinant active form of human CD154 (18‐kDa CD154). Human IgG production, alloimmunization, and in vitro T‐cell responsiveness were assessed.

RESULTS: There was no significant effect of 18‐kDa CD154 on human IgG levels in these mice, but it inhibited the development of HLA‐specific alloantibody in this model to five subsequent untreated white cell challenges. In vitro T‐cell proliferation in a mixed lymphocyte culture was also prevented by 18‐kDa CD154.

CONCLUSION: The recombinant protein 18‐kDa CD154 inhibited the ability of the Hu‐PBL‐SCID mice to mount a secondary immune response to allostimulation. This implies that transfusion‐induced alloimmunization utilizes CD40‐CD154 co‐stimulation and that blockade of this pathway can inhibit T‐cell function and interfere with the development of alloimmunization.
Analysis of donor return behavior
Authors: Ownby H.E. , Kong F. , Watanabe K. , Tu Y. , Nass C.C. ,   for the Retrovirus Epidemiology Donor Study
Published: March 2003

BACKGROUND:Efforts to provide a safe, adequate blood supply have been inhibited by persistent shortages attributed to a lack of motivation on the part of the general public and inefficiency in recruiting processes. This study examined whether frequency of donations and/or timing of subsequent donations by first‐time donors related to donor demographics.

STUDY DESIGN AND METHODS:Characteristics of 879,816 first‐time donors making at least one whole‐blood donation were analyzed. Cox proportional‐hazards regression models evaluated the first 10 return times separately, and a recurrent‐event Cox model was applied to simultaneously evaluate the first five returns.

RESULTS:The shorter the donation interval between the first two donations, the more likely the donor was to make subsequent donations. The proportion of repeat donors increased with education level. Rate of donation increased with age and education. The recurrent‐event Cox regression model showed that Rh‐negative donors, older donors, and donors who had completed college had higher donation return rates.

CONCLUSION:Time to return for second donation was associated with total number of donations made and with return rate for subsequent returns. Age was the strongest predictor of high donation frequency and early‐return rate. Relationships between interdonation interval and the number of future donations may prove useful in understanding return behavior and developing donor recruitment and retention strategies.
Direct HIV testing in blood donations: variation of the yield with detection threshold and pool size
Authors: Le Corfec E. , Le Pont F. , Tuckwell H.C. , Rouzioux C. , Costagliola D.
Published: March 2003

BACKGROUND: This study was designed to evaluate the potential yield of introducing nucleic acid amplification testing (NAT) in blood donation, according to the detection threshold and the pool size.

STUDY DESIGN AND METHODS: A mathematical model of early HIV‐1 population dynamics in blood has been developed and is used to predict the window period for NAT, according to the detection threshold and the pool size. The corresponding number of undetected, infected blood donations and the residual risk are estimated by using a previously published simulation model for the United States (9.96 million blood donations from regular donors, and an observed rate of HIV antibody‐positive blood donations of 3.18/100,000) and for France (2.32 million blood donations, and a rate of antibody‐positive donations of 0.9/100,000).

RESULTS: The average window period from infection predicted by the mathematical model for NAT ranges from 8.4 to 15.6 days, according to the detection threshold and the pool size. The maximum yield of adding NAT to the current antibody tests is estimated at 14 donations for the United States and 2 for France. The maximum yield of adding NAT to the newly developed combined HIV antibody and p24 antigen tests is 7 donations for the United States and 1 for France.

CONCLUSION: NAT at blood donation could reduce the HIV‐1 window period to a minimal value of 8 days without pooling the blood samples, but the yield of NAT would be close to that of combined HIV antibody and p24 antigen tests for high values of the detection threshold and the pool size.
Transfusion‐acquired hepatitis C: the Danish lookback experience
Authors: Christensen Peer B. , Groenbæk Karin , Krarup Henrik B. , Gaub Johannes , Møller Axel , Skinhøj Peter , Ring‐Larsen Helmer , Groenbæk Karin , Krarup Henrik B. , Dickmeiss Dickmeiss , Wantzin Per S.
Published: March 2003

BACKGROUND: In 1996, the Danish National Board of Health recommended hepatitis C virus (HCV) lookback to identify recipients of blood components from donors found to be positive since the implementation of anti‐HCV screening in 1991.

STUDY DESIGN AND METHODS: The aim was to accumulate results of the lookback at a national level and to describe the morbidity of the infected recipients. Records of transfusion were identified for at least 10 years back, and recipients still alive were tested for hepatitis C. Those with positive results were referred for clinical evaluation.

RESULTS: A total of 150 anti‐HCV‐positive Danish donors had donated blood to 1018 transfusion recipients, of whom 288 (29%) were still alive. Because of age, malignancy, or other severe diseases, 118 (41 %) of these were not contacted. Of 157 recipients screened for HCV, 128 (82%) were anti‐HCV positive on enzyme‐linked immunosorbent assay, and 88 (56%) were HCV RNA positive. Among the HCV RNA‐positive recipients, symptoms were present in 38 percent (25/66 reported), elevated alanine aminotransferase was found in 53 percent (41/77 tested), and cirrhosis was found in 11 percent (6/54 biopsied). Treatment with interferon α was initiated in 23 patients, corresponding to 26 percent of HCV RNA positive recipients.

CONCLUSION: Among tested recipients in the Danish HCV lookback, most were anti‐HCV positive and more than half were still viremic. The morbidity was considerable, and one‐fourth of viremic recipients entered treatment.