bluuebunny/crossref_metadata_2025 huggingface dataset:
Inactivation by phthalocyanine photosensitization of multiple forms of human immunodeficiency virus in red cell concentrates
Authors: Margolis‐Nunno H. , Ben‐Hur E. , Gottlieb P. , Robinson R. , Oetjen J. , Horowitz B.
Published: March 2003

BACKGROUND: The use of phthalocyanines in conjunction with red light has been shown to inactivate model lipid‐enveloped viruses in red cell concentrates. The ability of this treatment to inactivate multiple forms of human immunodeficiency virus (HIV) was evaluated in this study. STUDY DESIGN AND METHODS: The phthalocyanines used were aluminum phthalocyanine tetrasulfonate (AIPcS4) and the silicon phthalocyanines HOSiPcOSi(CH3)2(CH2)3 N(CH3)2 (Pc 4), and HOSiPcOSi(CH3)2 (CH2)3N+(CH3)3I‐(Pc 5). HIV was studied in a cell‐free form, in an actively replicating form, in latently infected cells, and in blood from HIV‐positive patients. RESULTS: All three phthalocyanines inactivate > or = 10(5) infectious doses of cell‐free HIV. However, only Pc 4 effectively inactivated actively replicating HIV and latently infected cells. The latter was about four times as sensitive to inactivation as was actively replicating HIV. Increasing the hematocrit of red cells during treatment decreased the rate of inactivation, especially at lower light doses. Under treatment conditions that completely inactivated the laboratory isolates of HIV, cell‐associated HIV in blood from HIV‐positive patients was also completely inactivated. The polymerase chain reaction signal from the gag gene of HIV was not affected on treatment of cell‐free virus, but it was reduced after treatment of cell‐associated HIV, particularly in some latently infected cell lines. CONCLUSION: Pc 4 and red light are effective in eliminating the infectivity of HIV in red cell concentrates. The usefulness of this approach for blood banking depends on future demonstration of the preservation of red cell circulatory survival and function in vivo.
Value and cost‐effectiveness of screening blood donors for antibody to hepatitis B core antigen as a way of detecting window‐phase human immunodeficiency virus type 1 infections
Authors: Busch M. P. , Dodd R. Y. , Lackritz E. M. , AuBuchon J. P. , Birkmeyer J. D. , Petersen L. R.
Published: March 2003

BACKGROUND: The value of screening donors for antibody to hepatitis B core antigen (anti‐HBc) for the prevention of posttransfusion hepatitis has declined markedly. However, anti‐HBc screening may still be useful as a surrogate marker for the window period (WP) of human immunodeficiency virus type 1 (HIV‐1) infection. STUDY DESIGN AND METHODS: First, the relationship between anti‐HBc reactivity and HIV‐1 WP infections was examined among 225 donors who had seroconverted to anti‐HIV‐1 positivity between 1987 and 1990. In addition, data from 1654 HIV‐1 seropositive donors were analyzed to characterize the relationship among anti‐HBc reactivity, donor demographics, and HIV‐1‐ related risk factors. The yield and cost‐effectiveness of anti‐HBc for HIV‐1 prevention were then projected on the basis of a published decision analysis model. RESULTS: Forty (18%) of 225 HIV‐1‐ seroconverting donors tested anti‐HBc‐reactive on the donation preceding anti‐HIV‐1 seroconversion; in contrast, 341 (34%) of 1014 HIV‐ 1‐seropositive donors interviewed tested anti‐HBc‐reactive (chi‐square test; p < 0.001). Anti‐HBc reactivity was more common among HIV‐1‐ seropositive donors reporting male‐to‐male sexual contact (169/360, 47%) and injection drug use (44/83, 53%) than among those with heterosexual contacts known to be HIV‐1‐positive (31/190, 16%) or transfusion exposure (3/21, 14%) or among females with no identified risk factors (21/124, 17%). The estimates of 18 to 34 percent sensitivity for anti‐HBc in detecting HIV‐1 WP donations and a current rate of 1 in 676,000 HIV‐1 WP donations (after p24 antigen screening) suggest that continued use of anti‐HBc screening could result in the transfusion of 5 to 12 fewer HIV‐1‐infected units per year in the United States, which would add 19 to 48 quality‐adjusted years of life for the 3.5 million annual transfusion recipients at a cost of $992,020 to $2,345,000 per quality‐adjusted life‐year saved. CONCLUSION: The low yield and very poor cost‐effectiveness of anti‐HBc screening indicate that this test is not an effective screening test for HIV‐1 WP donations.
The expression of p‐selectin during collection, processing, and storage of platelet concentrates: relationship to loss of in vivo viability
Authors: Holme S. , Sweeney J.D. , Sawyer S. , Elfath M.D.
Published: March 2003

BACKGROUND: Recent studies suggested that platelet activation with surface expression of p‐selectin on stored platelets may be related to a loss of viability. At present, there has been no thorough investigation of the extent or significance of p‐selectin expression during the collection, processing, and storage of platelet concentrates (PCs) under various conditions.

STUDY DESIGN AND METHODS: Platelet surface expression of p‐selectin (CD62) was determined on fixed platelet samples using fluorescein‐conjugated monoclonal antibodies. Platelet viability was assessed by autologous transfusion of platelets stored for 5 days and labeled with either 51Cr or 111in.

RESULTS: Little (2–10%) platelet expression of p‐selectin was found in whole blood and platelet‐rich‐plasma preparations, whereas PCs showed a substantial increase in p‐selectin expression to levels of 20 to 30 percent. Both fresh PCs and those stored for 5 days, obtained with one cell separator (MCS, Haemonetics) showed substantially lower levels of p‐selectin expression than PCs from two other cell separators (Spectra, COBE, and CS‐3000 with TNX‐6, Baxter Healthcare). Exposure of platelets to EDTA, cold, or a pH below 6.2, conditions that are known to result in the loss of viability upon transfusion, produced substantial and irreversible p‐selectin expression. PCs with a pH of 6.2 to 6.8 (conditions in which no loss of viability has been demonstrated) also showed pronounced p‐selectin expression, which returned to control values after incubation at 37° C in plasma at pH 7.0 to 7.2. With storage under current conditions the in vivo studies (n = 61) demonstrated a rather poor correlation between p‐selectin expression and the percentage of recovery (r = −0.25) but a somewhat better correlation with survival (r = −0.42). Better correlations were observed with the extent of shape change, lactate, and hypotonic shock response.

CONCLUSION: These studies show that p‐selectin expression on the platelet surface is a predictor of platelet viability, although the extent of shape change and the hypotonic shock response may be more sensitive.
Preapheresis peripheral blood CD34+ mononuclear cell counts as predictors of progenitor cell yield
Authors: Benjamin R.J. , Linsley L. , Fountain D. , Churchill W.H. , Sieff C. , Cannon M.E. , Uhl L. , Gaynes L. , Antin J.H. , Wheeler C.
Published: March 2003

BACKGROUND: Peripheral blood progenitor cells, harvested by apheresis after mobilization, provide rapid hematologic recovery after high‐dose chemotherapy. However, because harvesting these cells is expensive and time‐consuming, there has been much interest in optimizing collection protocols. An investigation was made to determine whether, in this clinical setting, peripheral blood progenitor cell yields may be predicted from preapheresis progenitor cell counts, allowing the length of each procedure to be “fine tuned” to achieve specific target goals.

STUDY DESIGN AND METHODS: Preapheresis peripheral blood CD34+ cell and total colony‐forming cell counts were assessed before 78 peripheral blood progenitor cell collections from 13 consecutive patients were performed. Preapheresis counts were correlated with actual progenitor cell yields. Factors affecting this correlation were analyzed.

RESULTS: With the use of linear regression analysis preapheresis progenitor cell counts were found to correlate significantly but weakly with actual yields per kg of body weight per liter of blood processed (CD34+ cells: r = 0.43; colony‐forming cells: r = 0.56). Further analysis revealed two possible causes: 1) circulating progenitor cell concentrations fluctuate widely during harvest, which implies that preapheresis counts are not representative of actual concentrations during apheresis, and 2) the efficiency with which apheresis machines extract mononuclear cells varies greatly between procedures.

CONCLUSION: Preapheresis CD34+ and colony‐forming cell counts correlated poorly with subsequent yields in this clinical setting, which suggests that it is not practical to use such counts to predict with certainty the length of apheresis needed to achieve a target yield.
Large‐volume apheresis for the harvest of peripheral blood progenitor cells for autologous transplantation
Authors: Smolowicz A.G. , Villman K. , Tidefelt U.
Published: March 2003

BACKGROUND: The mobilization and harvest of a sufficient number of peripheral blood stem and progenitor cells for autologous transplantation is an important aspect of treatment in patients with certain hematologic and solid tumor disease. The level of CD34+ cells in peripheral blood is often used as a predictor of successful harvest. STUDY DESIGN AND METHODS: A total of 129 apheresis procedures in 38 patients have been investigated retrospectively to evaluate the possibility to predict the outcome by other measures, such as total treated blood volume (TBV) during the apheresis. RESULTS: No significant correlation was observed between the level of CD34+ cells per kg of body weight in collected apheresis components and the TBV in all 129 apheresis procedures. However, analysis of results from 22 apheresis procedures with TBV > 16 L (large‐volume apheresis) and with < 10 × 10(3) CD34+ cells per mL in the peripheral blood found a correlation between TBV and the number of CD34+ cells per kg of body weight in the collected component (R2 = 0.585, p = 0.005). In patients who underwent large‐volume apheresis (> 16 L) and who had < 10 × 10(3) CD34+ cells per mL in their peripheral blood, the number of CD34+ cells in the apheresis component was not correlated with that in the peripheral blood prior to harvest (R2 = 0.262, p = 0.1569). In the patients who underwent apheresis procedures with TBV < 16 L and who had > 20 × 10(3) CD34+ cells per mL in their peripheral blood, there was a correlation between the number of CD34+ cells in the component and the number of CD34+ cells in the peripheral blood (R2 = 0.800, p = 0.0000). However, there was not a correlation in this group between the number of CD34+ cells in the component and the TBV. There were no significant differences in the content of CD34+/CD33+ and CD34+/ HLA‐DR+ cells in the collected component in the two groups. CONCLUSION: TBV appears to be critical for the collection of a sufficient number of progenitor cells in patients with < 10 × 10(3) CD34+ cells per mL in peripheral blood.
A general method for concentrating blood samples in preparation for counting very low numbers of white cells
Authors: Szuflad P. , Dzik W.H.
Published: March 2003

BACKGROUND: To count extremely low levels of white cells (WBCs) in WBC‐reduced blood components, a larger volume of sample must be processed. The goal was to develop an all‐purpose method for concentrating the samples obtained from WBC‐reduced red cells or platelets. The method was designed to be compatible with a variety of counting techniques.

STUDY DESIGN AND METHODS: Coded samples of red cell concentrates with an expected WBC concentration of 200, 100, 50, and 10 per mL and of the diluent (undetectable WBCs/mL) were sent to three sites on five occasions and counted by the use of the concentration method, crystal violet stain, and a Nageotte counting chamber. Additional samples were tested by flow cytometry, polymerase chain reaction, and volumetric capillary cytometry.

RESULTS: The results from the three test sites showed good linearity, with an overall r2 = 0.9994. The lower limit of accurate detection of the assay was 10 WBCs per mL. The results were biased toward underestimation, particularly at one of the test sites (Site A). There were no significantly different results in Sites B and C. The intra‐assay CV was acceptable. Precision (reproducibility) at the three test sites varied.

CONCLUSION: This method allows reliable determination of WBC concentrations as low as 0.01 per μL in blood. Despite the use of technologists trained in Nageotte chamber counting, validation testing demonstrated that one test site's performance was significantly different from that of the other two sites, because of both underestimation bias and variation in count results. The sample concentration method, when used in conjunction with an automated assay for WBC identification, should permit larger volume analysis with a greater degree of precision and a lower limit of detection than is found in assays that do not concentrate the sample before counting.
Delayed hemolytic transfusion reactions in sickle cell disease: simultaneous destruction of recipients' red cells
Authors: King K. E. , Shirey R. S. , Lankiewicz M. W. , Young‐Ramsaran J. , Ness P. M.
Published: March 2003

BACKGROUND: Bystander hemolysis may be defined as the destruction of antigen‐negative red cells during immune hemolysis, such as delayed hemolytic transfusion reaction (DHTR). Although many have suspected that bystander hemolysis does occur, that phenomenon is very difficult to document. STUDY DESIGN AND METHODS: Five patients with sickle cell disease (SCD) who underwent exchange transfusion and subsequently experienced a DHTR were retrospectively evaluated. Serial samples were examined for complete blood counts, the percentage of hemoglobin A and S, and the percentage of reticulocytes. The total red cell count and the percentage of hemoglobin S were used to calculate the hemoglobin S red cell count. The patients' profiles were compared to proposed models. RESULTS: DHTRs due to anti‐E, ‐S, ‐Fy(a), or ‐Jk(a) or serologically undetectable antibodies were identified 7 to 19 days after exchange transfusion. All patients had a significant decrease in hemoglobin A red cells; 56.4 to 94.7 percent of hemoglobin A red cells were hemolyzed. Patients 4 and 5 had a decrease in hemoglobin S red cells which indicated the destruction of autologous red cells during DHTR. The evidence for bystander hemolysis was particularly convincing in Patient 5, because there was a substantial decrease in hemoglobin S red cells despite a reticulocyte production index of 2.2 percent at the nadir of the DHTR. CONCLUSION: Hemoglobin S provides a biologic marker for monitoring autologous red cell loss in sickle cell patients. We have shown one patient with clinical evidence of bystander hemolysis complicating a DHTR.
Prevalence of hepatitis G virus and its strain variant, the GB agent, in blood donations and their transmission to recipients
Authors: Roth W.K. , Waschk D. , Marx S. , Tschauder S. , Zeuzem S. , Bialleck H. , Weber H. , Seifried E.
Published: March 2003

BACKGROUND: Hepatitis G virus (HGV) and its strain variant, the GB agent (GBV‐C) are independent isolates of a recently identified non‐A through ‐E hepatitis virus. Prevalence in United States volunteer blood donors is 1.5 to 1.9 percent, but no data on European blood donors are available. Epidemiologic data suggest a preferred parenteral transmission route. The prevalence of HGV/GBV‐C in European blood donors and the efficiency of transmission to transfusion recipients were investigated. STUDY DESIGN AND METHODS: Plasma samples from unpaid volunteer German blood donors were tested for HGV/GBV‐C by in‐house reverse transcription‐polymerase chain reaction. Positive donors were independently retested and interviewed for parenteral transmission risks. Amplification products were sequenced and subjected to phylogenetic analysis. Recipients of reverse transcription‐polymerase chain reaction‐positive donations were traced and tested for HGV/GBV‐C infection. RESULTS: A total of 14 (1.34%) of 1048 donors (alanine aminotransferase < 45 IU/L) were repeatedly positive for HGV/GBV‐C with 9 (2.18%) of 413 urban and 5 (0.78%) of 635 rural donors (chi 2‐test; p = 0.04). Isolates differed in nucleotide sequence homology over a range of 12.5 to 19.6 percent. All but one positive donor reported parenteral transmission risks. Transmission of HGV/GBV‐C was detected in 4 of 9 transfusion recipients. The prevalence of HGV/GBV‐C in donors with an alanine aminotransferase level > 45 IU per L was 3 percent (3/100). Two mother/child pairs were identified with highly homologous isolates. CONCLUSION: A significantly greater prevalence of HGV/GBV‐C was detected in urban volunteer blood donors than in rural donors. The high prevalence in urban donors (2.18%) suggests specific transmission risks for this group. The less than 50‐percent efficiency of HGV/GBV‐C transmission via blood components may indicate the presence of defective viruses with reduced infectivity. There is evidence for vertical transmission.
Plateletpheresis in 90‐ to 110‐pound donors using the CS‐3000 blood cell separator
Authors: Buchholz D.H. , Squires Jerry E. , Herman Jay H. , Ng Ambrose T. , Anderson Joy K. , Hedberg Sandra L.
Published: March 2003

BACKGROUND: Increases in the use of single‐donor apheresis components have increased the need for platelet donors. In the United States, persons must weigh 110 pounds or more to qualify as blood donors, and the same weight limitation has been placed on apheresis donors. Because automated plateletpheresis with some instruments differs considerably from whole‐blood donation with respect to the volume of blood removed from the donor, the feasibility of using persons weighing between 90 and 110 pounds as platelet donors was evaluated by the use of the CS‐ 3000 blood cell separator. STUDY DESIGN AND METHODS: The study was performed using female subjects who met all usual donor requirements except for minimum weight. The standard platelet collection procedure of the instrument was used, except that the blood processing rate was manually selected so as to optimize the blood withdrawal and return rate in individuals. Vital signs were recorded before and after donation as were signs or symptoms of any type of donor reaction. RESULTS: Twenty‐six of 28 women completed the donation procedure; in two instances, collection was terminated prematurely because of an inability to maintain adequate venous access. An average of 4.5 × 10(11) platelets were collected during a mean donation time of 110 minutes. All donors tolerated the procedure well, and no serious adverse reactions were seen. Because of the administration of priming solution and anticoagulant during apheresis, there was a net positive fluid balance following the procedure, in spite of the removal of approximately 220 mL of platelet concentrate. CONCLUSION: These preliminary studies suggest that 90‐ to 110‐pound persons may serve as plateletpheresis donors. Additional studies are needed to more fully document the safety and efficacy of this approach. The use of lower‐ weight donors may significantly increase the number of persons available to provide single‐donor platelet components.
Chronic anemia due to parvovirus B19 infection in a bone marrow transplant patient after platelet transfusion
Authors: Cohen B.J. , Beard S. , Knowles W.A. , Ellis J.S. , Joske D. , Goldman J.M. , Hewitt P. , Ward K.N.
Published: March 2003

BACKGROUND: Many reports document the transmission of human parvovirus B19 (B19) infection by clotting factor concentrates manufactured from large plasma pools. Transmission via other blood components originating from a single donor or a small number of donors, however, seems to occur only rarely. The study reported here identifies a B19 infection that was transmitted via a platelet donation. CASE REPORT: A multiply transfused allogeneic bone marrow transplant patient developed chronic anemia due to persistent B19 infection. The anemia responded to therapy with intravenous immunoglobulin. It was postulated that a transfusion was the source of the B19 infection. Archived sera from 90 implicated blood donors were tested for B19 IgM and DNA by the use of dot‐blot hybridization and a nested polymerase chain reaction with primers from the B19 nonstructural gene. B19 DNA from patient and donor sera were sequenced. One of the 90 blood donors (Donor A) was B19 IgM positive and had a high level of B19 DNA. The patient was viremic 3 days after transfusion of platelets from this donor, and the sequence of B19 DNA from the patient exactly matched that of B19 DNA from the donor. A second blood donor (Donor B) had a low level of B19 DNA but was IgM negative. The patient showed no evidence of B19 infection after the transfusion of red cells from Donor B, and the sequence of this donor's B19 DNA was different from that in the patient. CONCLUSION: Blood Donor A with asymptomatic acute B19 infection was the source of B19 infection in the bone marrow transplant patient. Donor B with a low level of B19 DNA was not the source of infection.
The specific hospital significantly affects red cell and component transfusion practice in coronary artery bypass graft surgery: a study of five hospitals
Authors: Surgenor D.M. , Churchill W.H. , Wallace E.L. , Rizzo R.J. , McGurk S. , Goodnough L.T. , Kao K.J. , Koerner T.A. , Olson J.D. , Woodson R.D.
Published: March 2003

BACKGROUND: Interhospital differences in blood transfusion practice during coronary artery bypass graft (CABG) surgery have been noted, but the underlying issues have not been identified. STUDY DESIGN AND METHODS: Records of 3217 consecutive CABG cases in five university teaching hospitals in 1992 and 1993 were stratified by hospital, type of revascularization conduit, patients' sex, and other factors. Statistical methods were used to compare patient characteristics, transfusion outcomes, and hospital outcomes. RESULTS: Forward two‐step logistic regression using patient likelihood of red cell transfusion factors in the first step and the specific hospital in the second step revealed a significant effect of hospital on the delta odds ratios for red cell transfusion. This finding was confirmed by analyses of a highly stratified subset of cases, males in diagnosis‐related group 107 (primary cases of coronary bypass without coronary catheterization) who underwent revascularization with venous and internal mammary artery grafts, revealing variations among hospitals from 109 to 457 units of red cells transfused per hundred cases. Corresponding variations in transfusions of all blood components were from 324 to 1019 units by hospital. Variation in red cell transfusion practice among surgeons in the same hospital was not responsible for these interhospital differences. CONCLUSION: The effect of the specific hospital on transfusion practice is attributed to institutional differences that, through reasons of training or hierarchy, become ingrained in hospitals.
The relationship between the duration of platelet storage and the development of transfusion reactions
Authors: Sarkodee‐Adoo C.B. , Kendall J.M. , Sridhara R. , Lee E.J. , Schiffer C.A.
Published: March 2003

BACKGROUND: The incidence of platelet transfusion reactions may depend partly on the length of storage. The influence of reactions on the effectiveness of platelet transfusions is not known. STUDY DESIGN AND METHODS: Platelet transfusion reactions, identified by prospective monitoring, were analyzed for the effects of component type, recipient lymphocytotoxic antibodies, bacterial contamination, and duration of storage. Posttransfusion corrected count increments (CCIs) were used to evaluate the effectiveness of transfusions associated with reactions by comparing them to those of randomly selected transfusions without reactions. RESULTS: Reactions accompanied 4 percent of the 4926 transfusions given and included 119 febrile nonhemolytic transfusion reactions, 62 allergic reactions, and 13 reactions with features of both. Platelet concentrates contained a mean of 0.5 × 10(8) white cells per unit. Lymphocytotoxic antibodies were detectable in 20 of 84 recipients tested proximate to a reaction. Bacterial cultures from 4 of 81 units were positive; 1 unit was associated with fatal Enterobacter sp. sepsis. The incidence of febrile nonhemolytic transfusion reactions but not allergic reactions was related to platelet storage duration. The CCI was not significantly different for transfusions associated with reactions (10.97 [median, range 0–72.5; n = 165]) or not so associated (13.1 [median, range 0–39.5; n = 174]) (p = 0.08). CONCLUSION: The incidence of febrile nonhemolytic transfusion reactions but not allergic reactions appears to be related to the duration of platelet storage. Transfusion reactions may not have an adverse impact on the effectiveness of platelet transfusions.
Detection of fetal red cells in fetomaternal hemorrhage using a fetal hemoglobin monoclonal antibody by flow cytometry
Authors: Davis B.H. , Olsen S. , Bigelow N.C. , Chen JC.
Published: March 2003

BACKGROUND: The laboratory determination of the level of fetal cells in maternal circulation remains an important support in the obstetrical management of women with suspected uterine trauma and in the proper dose administration of anti‐D for prevention of Rh hemolytic disease of the newborn. Limitations in the sensitivity and precision of the widely used manual Kleihauer‐Betke test have prompted an increased utilization of flow cytometric methods for fetal cell detection in maternal blood samples.

STUDY DESIGN AND METHODS:Murine monoclonal antibodies directed against fetal hemoglobin (HbF) were developed, conjugated to fluorescein isothiocyanate, and used in a multiparametric flow cytometric assay developed for the quantitation of fetal red cells. A rapid intracellular staining method using brief glutaraldehyde fixation and Triton X‐100 permeabilization prior to monoclonal antibody incubation was developed, along with optimization of the flow cytometric analysis protocol for the analysis of 50,000 cells. The performance of the assay was assessed for linearity and precision and correlated with the Kleihauer‐Betke acid elution method.

RESULTS:The anti‐HbF flow cytometric method showed good correlation with the Kleihauer‐Betke method (r2 = 0.86) and superior precision with a CV < 15 percent for blood samples with > 0.1 percent fetal cells. Analysis of 150 blood samples from nonpregnant adults, including individuals with elevated HbF due to hemoglobinopathies and hereditary persistence of HbF, gave a mean value of 0.02 percent fetal cells, and all results were less than 0.1 percent.

CONCLUSIONS:The anti‐HbF flow cytometric method for detection of fetal cells offers a simple, reliable, and more precise alternative to the Kleihauer‐Betke manual technique for the assessment of fetomaternal hemorrhage. The method has additional potential applications for the study of HbF levels or frequency of adult red cells with low levels of HbF (F cells) in individuals with hemoglobinopathies.
A prospective randomized trial of the surgical blood order equation for ordering red cells for total hip arthroplasty patients
Authors: Nuttall G. A. , Santrach P. J. , Oliver W. C. , Ereth M. H. , Horlocker T. T. , Cabanela M. E. , Trousdale R. T. , Bryant S. , Currie T. W.
Published: March 2003

BACKGROUND: The majority of crossmatched blood is for surgical patients, and most of it is never transfused. An alternative system for ordering red cell (RBC) units, called the surgical blood order equation (SBOE), which incorporates specific patient variables for surgical patients, has been developed.

STUDY DESIGN AND METHODS: A prospective double‐blind randomized trial compared the SBOE with the maximal surgical blood order schedule (MSBOS) system for ordering allogeneic RBC units in 60 patients undergoing total hip arthroplasty. Autologous RBCs were available for none of the patients.

RESULTS: There were no differences in patient demographic, surgical, or laboratory variables at any time. The median number (range) of allogeneic RBC units ordered was 2 (2–3) for the MSBOS and 0 (0–3) for the SBOE (p<0.0001). The SBOE ordered the correct number of RBC units for 58 percent of patients, while the MSBOS did so for 7 percent (p<0.0001). The SBOE had a lower crossmatch‐to‐transfusion ratio than the MSBOS (0.83 vs. 4.12). Costs were also lower with the SBOE.

CONCLUSION: Incorporation of patient factors in the use of the SBOE system resulted in increased efficiency of blood‐ordering practices for total hip arthroplasty.
The predictive value of white cell or CD34+ cell count in the peripheral blood for timing apheresis and maximizing yield
Authors: Yu J. , Leisenring W. , Bensinger W.I. , Holmberg L.A. , Rowley S.D.
Published: March 2003

BACKGROUND: The collection of peripheral blood stem and progenitor cells (PBPCs) for transplantation can be time‐consuming and expensive. Thus, the utility of counting CD34+ cells and white cells (WBCs) in the peripheral blood was evaluated as a predictor of CD34+ cell yield in the apheresis component.

STUDY DESIGN AND METHODS: The WBC and CD34+ cell counts in the peripheral blood and the apheresis components from 216 collections were assessed. Sixty‐three patients underwent mobilization with chemotherapy plus filgrastim, and 17 patients and 14 allogeneic PBPC donors did so with filgrastim alone. The relationship between the number of WBC and CD34+ cells in the peripheral blood and in the apheresis component was analyzed by using rank correlation and linear regression analysis.

RESULTS: The correlation coefficient for CD34+ cells per liter of peripheral blood with CD34+ cell yield (× 106/kg) was 0.87 (n = 216 collections). This correlation existed for many patient and collection variables. However, patients with acute myeloid leukemia had fewer CD34+ cells in the apheresis component at any level of peripheral blood CD34+ cell count. Components collected from patients with CD34+ cell counts below 10 × 106 per L in the peripheral blood contained a median of 0.75 × 106 CD34+ cells per kg. When the WBC count in the blood was below 5.0 × 109 per L, the median number of CD34+ cells in the peripheral blood was 5.6 × 106 per L (range, 1.0‐15.5 × 106/L). A very poor correlation was found between the WBC count in the blood and the CD34+ cell yield (p = 0.12, n = 158 collections).

CONCLUSION: The number of CD34+ cells, but not WBCs, in the peripheral blood can be used as a predictor for timing of apheresis and estimating PBPC yield. This is a robust relationship not affected by a variety of patient and collection factors except the diagnosis of acute myeloid leukemia. Patients who undergo mobilization with chemotherapy and filgrastim also should undergo monitoring of peripheral blood CD34+ cell counts, beginning when the WBC count in the blood exceeds 1.0 to 5.0 × 109 per L.
The effect of unmodified or prestorage white cell‐reduced allogeneic red cell transfusions on the immune responsiveness in orthopedic surgery patients
Authors: Bordin J.O. , Chiba A.K. , Carvalho K.I.L. , Takata E.T. , Falcão R.P. , Garcia A.B. , Bordin I.A.S. , Maciel M.M. , Andreoni S. , Kerbauy J.
Published: March 2003

BACKGROUND: The immunomodulatory effects of allogeneic blood transfusions have been attributed to the white cells (WBCs) present in the cellular blood components transfused to patients.

STUDY DESIGN AND METHODS: The effect of the transfusion of allogeneic red cells (RBCs) or allogeneic prestorage WBC‐reduced RBCs (WBC‐reduced RBCs) on host immune responsiveness was evaluated by measuring the lymphocyte subsets and the in‐vitro cytokine production in response to phytohemagglutinin stimulation of WBCs of orthopedic surgery patients. Forty‐seven patients undergoing hip replacement surgery were randomly assigned to receive allogeneic RBCs (n = 17) or WBC‐reduced RBCs (n = 14; 99.95% WBC removal). Sixteen patients were not transfused. Patient blood samples taken before surgery and on Days 1 and 4 after surgery were tested for complete blood count, lymphocyte subset analysis, and measurement of cytokine levels.

RESULTS: After surgery, the lymphocyte count was significantly decreased in patients transfused with ≥3 units of allogeneic RBCs (2.0 ± 0.5 vs. 1.3 ± 0.3 × 109/L; p = 0.017), but not in patients transfused with ≥3 units of WBC‐reduced RBCs (2.0 ± 0.9 vs. 1.7 ± 0.8 × 109/L). Compared with preoperative levels, on Day 4 after surgery, patients transfused with ≥3 units of allogeneic RBCs also had a decrease in the number of natural killer cells (0.07 ± 0.05 vs. 0.04 ± 0.03 × 109/L; p = 0.018). Postoperatively, interleukin‐2 was decreased in one patient who received WBC‐reduced RBCs compared with that in four patients transfused with allogeneic RBCs (p = 0.32), and eight untransfused patients (p = 0.01). On Day 4, about 70 percent of patients transfused with allogeneic RBCs showed a 20‐percent decrease in the interferon gamma level.

CONCLUSION: Taken together, these data support the hypothesis that transfusion of ≥3 units of allogeneic RBCs is associated with early postoperative lymphopenia in otherwise healthy individuals undergoing surgery. These findings were not observed in those individuals transfused with RBCs that had undergone prestorage WBC reduction.
Antibodies to private and public HLA class I epitopes in platelet recipients
Authors: Zimmermann R. , Wittmann G. , Zingsem J. , Blasczyk R. , Weisbach V. , Eckstein R.
Published: March 2003

BACKGROUND: Transfusions or pregnancies can cause immunization against private HLA determinants and public epitopes shared by more than one private HLA antigen. HLA antibodies are correlated with febrile transfusion reactions, lower platelet response following platelet transfusion, and an increased rate of renal transplant rejection. Until now, antibody specificities in alloantisera from platelet recipients have been poorly characterized.

STUDY DESIGN AND METHODS: Consecutive serum screens from platelet recipients were analyzed for antibodies against private HLA class I antigens and public HLA epitopes using a serum analysis program based on the 2 × 2 table analysis of correlations. Serum screens of highly immunized patients and of patients with new alloimmunization events were reviewed separately.

RESULTS: Of the serum screens from 566 platelet recipients, 1577 indicated alloimmunization (panel‐reactive antibodies >5%). The program assigned a specificity in 1024 of these screens (64.9%) and at least once in 522 of 566 patients (92.2%). In 267 patients, antibodies detecting public epitopes in the combined A‐ or B‐locus cross‐reacting groups were found; other public markers were detected in 39 patients. Patterns of reactivity were remarkably less stable than in patient groups previously studied. In many patients, antibodies with apparent private epitope specificity preceded the identification of antibodies against a shared marker of the same cross‐reactive group. However, the disappearance of antibodies (whether or not this was followed by a new antibody against a private or public marker belonging to another cross‐reacting group) was also observed.

CONCLUSION: The computerized analysis of microlymphocytotoxicity tests enhances the rate of antibody specification in sera from platelet recipients with lymphocytotoxic antibodies. The identified antibodies should be taken into account in the selection of platelet donors. The data confirm and extend previous observations on HLA class I antibodies and elucidate new alloimmunization events.
Simultaneous extraction of hepatitis C virus (HCV), hepatitis B virus, and HIV‐1 from plasma and detection of HCV RNA by a reverse transcriptase‐polymerase chain reaction assay designed for screening pooled units of donated blood
Authors: Sun R. , Schilling W. , Jayakar H. , Ku J. , Wang J. , Rosenstraus M. , Spadoro J.
Published: March 2003

BACKGROUND: Testing for viral nucleic acids should reduce the residual risk of transmitting viral infections by transfusion of blood components. The AmpliScreen Hepatitis C (HCV) Test, Version 2.0, was designed for screening pools composed of samples from individual units of blood or plasma.

STUDY DESIGN AND METHODS: An ultracentrifugation step during sample processing simultaneously extracts and concentrates HCV, HIV type 1, and hepatitis B virus particles from plasma. An HCV internal control RNA serves as an extraction and amplification control for each processed sample. Processed samples are amplified by reverse transcriptase polymerase chain reaction and detected by hybridization of the amplified products to HCV‐ and internal‐control‐specific oligonucleotide probes. Plasma samples containing known quantities of HCV were used to evaluate analytical sensitivity and precision.

RESULTS: The analytical sensitivity of the test was 25 IU of HCV per mL of pooled plasma; all HCV genotypes were detected with similar efficiency. The test did not react with other blood‐borne viruses. The within‐run and total coefficients of variation were 1.3‐13.0 percent and 1.7‐22.0 percent, respectively, with low copy samples producing the more variable results. The test performed well using plasma collected in either EDTA, ACD, or PPT Vacutainer tubes. Plasma samples containing elevated levels of hemoglobin, albumin, triglycerides, or bilirubin did not interfere with the test. The test detected HCV RNA 23 to 32 days prior to seroconversion for four of the five seroconversion panels tested.

CONCLUSION: The AmpliScreen HCV Test, Version 2.0 provides a reproducible and specific method for screening pooled blood units for HCV. Theoretically, this test has sufficient sensitivity to detect a single infected unit containing 2.4 × 103 IU of HCV per mL in a pool with 95 uninfected units and should reduce the window period by at least 20 to 30 days.
Biotinylation modifies red cell antigens
Authors: Cowley Helen W. , Wojda Urszula , Cipolone Karen M. , Procter Jo L. , Stroncek David F. , Miller Jeffery L.
Published: March 2003

BACKGROUND: Chemical biotinylation of red cell membranes may be useful for several clinical applications, including red cell survival studies.

STUDY DESIGN AND METHODS: To examine the possible effects of biotinylation on red cell antigens, standard hemagglutination assays were performed on matched sets of control and biotinylated red cells. The red cells were biotinylated at a final concentration of 2.0 pg of sulfo‐N‐hydroxysuccinimide‐biotin per cell, and antigen‐negative cells were directly compared to antigen‐positive cells when possible. The hemagglutination assays were graded in a blinded fashion. Forty‐one red cell antigens from 21 of the 23 established blood group systems were tested.

RESULTS: Hemagglutination based upon antibody binding to A, A1, M, N, S, s, P1, D, C, E, c, e, Cw, Lub, K, k, Kpb, Lea, Leb, Fya, Fyb, Jka, Jkb, Dia, Wra, Wrb, Yta, Xga, Sc1, Dob, Coa, Ch, H, Ge2, Cra, Kna, I, and P was not affected by biotinylation. Unexpectedly, the hemagglutination of Di(b+) and LW(a+) red cells was blocked after biotinylation. Conversely, MH04 monoclonal anti‐A agglutinated red cells expressing B only after biotinylation. BIRMA‐1 monoclonal anti‐A and polyclonal anti‐A from sera did not agglutinate the biotinylated B red cells.

CONCLUSION: Biotinylation of human red cells specifically modified their antigenicity, as measured by standard hemagglutination assays.
Preoperative autologous blood donation by 1073 elderly patients undergoing elective surgery: a safe and effective practice
Authors: Gandini Giorgio , Franchini Massimo , Bertuzzo Daniela , Olzer Daniela , Crocco Isabella , De Gironcoli Marzia , Aprili Giuseppe
Published: March 2003

BACKGROUND: Preoperative autologous blood donation (PABD) aims at avoiding the risks associated with exposure to allogeneic blood. While its use is extremely common among adult patients in connection with elective surgery, it is still uncommon in elderly patients, because of a series of coexisting pathologies.

STUDY DESIGN AND METHODS: A retrospective study was made of 1073 consecutive elderly patients at a city hospital from 1990 to 1996. Their responses to the PABD program were evaluated by analysis of the incidence of complications and the demand for allogeneic blood.

RESULTS: The PABD program was interrupted in 79 (7.4%) of 1073 patients because of the onset of anemia, vasovagal reactions, lack of accessible superficial veins, or cardiovascular complications. Seven hundred eightyfour (73.1%) of 1073 patients were given autologous blood; 151 (14.1%) patients also required allogeneic blood.

CONCLUSION: The onset of anemia (6.5%) was the main contraindication for continuing the PABD program: incidence increased with age. PABD in connection with elective surgery is both feasible and effective in a high percentage of elderly patients.
Genomic characterization of the Kidd blood group gene:different molecular basis of the Jk(a–b–) phenotypein Polynesians and Finns
Authors: Irshaid N.M. , Henry S.M. , Olsson M.L.
Published: March 2003

BACKGROUND: The clinically important Kidd (JK) blood group antigens are carried by the urea transporter in red cells. The rare Jk(a–b–) phenotype can be caused by homozygosity at the JK locus for a silent allele, Jk. This phenotype has been recorded in many ethnic groups, but it is most abundant among people originating from the Polynesian Islands and Finland. The molecular basis for Jk(a–b–) is unknown in these populations.

STUDY DESIGN AND METHODS: Blood samples from individuals of Swedish, Polynesian, and Finnish origin were collected and characterized by routine JK blood group serology and JK genotyping. Genomic DNA covering the exons and intervening introns of the JK gene coding region was amplified by polymerase chain reaction, and fragments were directly sequenced.

RESULTS: Exon and partial intron sequences in the coding region of the JK gene were determined. Finnish and Polynesian Jk alleles were analyzed; the only deviations from consensus were a splice‐site mutation (G→A) in Polynesians, causing skipping of exon 6, and a T871C substitution predicted to disrupt a potential N‐glyco‐sylation motif (NSS→NSP) in Finns. Methods for rapid detection of silent Jk alleles were developed for clinical application.

CONCLUSION: Polynesians and Finns have two different molecular alterations in their Jk alleles, both of which can now be determined by polymerase chain reaction.
White cell apoptosis in platelet concentrates
Authors: Frabetti F. , Tazzari P.L. , Musiani D. , Bontadini A. , Matteini C. , Roseti L. , Tassi C. , Viggiani M. , Marini M. , Conte R.
Published: March 2003

BACKGROUND: The aim of the present study was the evaluation of the apoptosis in residual white cells (WBCs) contained in platelet concentrates (PCs) and of the relationship of this apoptosis with the concentration of inflammatory cytokines in the medium and with platelet activation.

STUDY DESIGN AND METHODS: Three independent methods were used to evaluated apoptosis in WBCs present in 9 PCs, either from single donors by apheresis (SD‐PCs) or from pooled buffy coats (BC‐PCs). All PCs were divided in two parts, one of which was irradiated. PCs were stored up to 4 days at room temperature, and samples were withdrawn daily for analysis of apoptosis, of platelet activation (surface and soluble CD62P), and of cytokine concentration (interleukin [IL]‐1α, IL‐1β, IL‐6, IL‐8, and tumor necrosis factor α).

RESULTS: Apoptosis was found to occur with storage in both irradiated and nonirradiated units. Platelet activation increased with storage time and was higher in BC‐PCs. The amount of released cytokines was rather variable among PC units. Only IL‐8 was consistently found to increase with storage time.

CONCLUSIONS: Apoptosis of residual WBCs occurred in PC units as a function of storage time. The amount and the time course of apoptosis seem to correlate with IL‐8 release rather than with platelet activation or with the occurrence of febrile nonhemolytic transfusion reactions.
Molecular mechanisms that lead to reduced expression of Duffy antigens
Authors: Yazdanbakhsh K. , Rios M. , Storry J.R. , Kosower N. , Parasol N. , Chaudhuri A. , Reid M.E.
Published: March 2003

BACKGROUND: In the Duffy blood group system, the null phenotype Fy(a–b–) has been classically associated with a mutated GATA box, while the Fyx phenotype weak Fyb is associated with Arg89Cys and Ala100Thr mutations. This report assesses the prevalence of the Duffy GATA box and the Fyx‐associated mutations in white and African American (black) donors and investigates the molecular mechanism underlying the Fyx phenotype.

STUDY DESIGN AND METHODS: PCR RFLP Duffy genotyping was performed on blood samples from blacks and whites. Duffy antigen expression (Fya, Fyb, Fy6, Fy3) on RBCs was measured by flow cytometry. By site‐directed mutagenesis, the relevance of each Fyx‐associated mutation to Duffy (mRNA, antigen, and protein) expression was analyzed in transfectants by Northern blotting, flow cytometry, and immunoblotting.

RESULTS: The mutated GATA box occurred at a high allele frequency (0.8) in blacks and was rare among whites. Conversely, the Fyx‐associated mutations were absent in blacks, but present in 3.5 percent of whites. By flow cytometry, Duffy antigens (Fya or Fyb, Fy6 and Fy3) showed a dosage effect in RBC samples that were transcriptionally silenced by the GATA box mutation in one allele. By contrast, the reduced (10%) Duffy protein in Fyx RBCs was shown by heterologous expression analysis not to be due to reduced RNA levels, but to protein instability caused by Arg89Cys.

CONCLUSIONS: R educed Duffy expression can result from mutations affecting transcription (mutated GATA box in one allele) or instability of the translated protein (Arg89Cys). The frequencies of these mutations vary among populations.
Identification of a novel hybrid glycophorin gene encoding GP.Hop
Authors: Storry J.R. , Poole J. , Condon J. , Reid M.E.
Published: March 2003

BACKGROUND: The GP.Hop (Mi.IV) phenotype expresses the MNS low‐incidence antigens Mur, Hop, TSEN, MINY, and MUT. Because serologically similar MNS phenotypes expressing some or all of these antigens were shown to be carried by hybrid GP(B‐A‐B) proteins, it was proposed that a similar protein would be found for GP.Hop. The identification of a second GP.Hop propositus (ES) initiated a study to determine the molecular basis of this phenotype.

STUDY DESIGN AND METHODS: Serologic tests and immunoblotting analysis with glycophorin‐specific antibodies were performed. GYPB, the gene encoding the GPB protein, was cloned and sequenced after reverse transcription PCR amplification of total RNA isolated from ES. GYPB‐specific primers encompassing GYPB pseudoexon 3, intron 3, and exon 4 were also used to clone and sequence genomic DNA from ES and MH, the original GP.Hop proband.

RESULTS: Serologic and immunochemical data confirmed that ES's RBCs carried antigens associated with the GP.Hop phenotype. Sequencing of ES's cDNA demonstrated the presence of genes predicted to encode s‐specific GPB and an S‐specific GP(B‐A‐B) hybrid in which the 3′ end of GYPB pseudoexon 3 had been replaced by a short nucleotide sequence from exon 3 of the GPA gene (GYPA). The hybrid nucleotide sequence contained sequence motifs previously shown to be required for the expression of the Mur, Hop, TSEN, MINY, and MUT, which is consistent with their presence as detected serologically. Genomic DNA analysis found that the crossover point in GYPB pseudoexon 3 was identical in ES and MH.

CONCLUSIONS: The GP.Hop phenotype is produced by a hybrid GP(B‐A‐B) protein caused by a DNA insertion of GYPA into GYPB. The composition of the hybrid protein is GPB(1‐26)‐GPψB(27‐50)‐GPA(51‐58)‐GPBS(59‐103).