bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of fibronectin‐containing blood products on lymphocyte reactivity
Authors: Schultz John C. , Shahidi N.T.
Published: March 2003

Two fibronectin (FN)‐containing blood products, human peripheral blood plasma and cryoprecipitate, were examined for their effect on mitogen‐ induced lymphocyte transformation in vitro. Responses of human peripheral blood lymphocytes to phytohemagglutinin (PHA) were depressed in the presence of a plasma concentration above that required for maximum DNA synthesis, and this concentration must be present in cultures prior to lymphocyte activation. The removal from the plasma of heparin‐induced cryoprecipitate, a complex consisting of FN, heparin, and fibrinogen, resulted in a significant reduction in the inhibitory effect of the plasma on the PHA response. Plasma specifically depleted of FN by affinity chromatography on gelatin‐agarose beads was 32 percent less inhibitory to the PHA‐induced stimulation of cells than untreated plasma; the remaining inhibitory activity in the FN‐depleted plasma samples was attributed to the presence of other normal immunosuppressive factors. The inhibitory capacity of FN in plasma was similar to that obtained with purified FN alone, which indicates that, unlike that of other known plasma inhibitors, the immunosuppressive activity of FN was not altered by the presence of other components of plasma. Cryoprecipitate used in the treatment of hemophilia contains high levels of FN, and, as anticipated, PHA‐induced lymphocyte transformation was markedly depressed in the presence of solubilized cryoprecipitate. The contribution of FN to the T‐cell abnormalities in patients chronically receiving cryoprecipitate and/or factor VIII concentrates derived from cryoprecipitate warrants further investigation.
Detection of cytomegalovirus antibody in platelet concentrates by fluorescence immunoassay and latex agglutination
Authors: DePalma L. , Criss V. R. , Sullivan M. T. , Leitman S. F. , Williams A. E. , Luban N. L.
Published: March 2003

A passive latex agglutination (PLA) test for cytomegalovirus (CMV) antibody detection has been shown to be an acceptable method of screening both donor sera and plasma from units of red cells and platelets stored in CPDA‐1. However, most plateletpheresis concentrates are collected in ACD, and CMV antibody testing of ACD‐stored products has not been systematically evaluated by PLA. Sera and ACD‐stored platelet concentrate bag segments from 104 donors were tested by PLA and by a solid‐phase fluorescence immunoassay (FIAX) as a reference standard for CMV‐IgM and CMV‐IgG antibodies. Sera were stored at both 4 and 22 degrees C and were tested on Days 1 and 5 of storage; segments were tested daily for 5 days. Of 63 donor samples (61%) that tested negative for CMV‐IgG by FIAX, there were two false‐positive results in bag segments by PLA testing, one on Day 1 and the other on Day 2 of storage. PLA testing was consistently positive in sera and segments in the 40 donors (38%) who tested positive for CMV‐IgG by FIAX. Potential false‐negative PLA results occurred in five bag segments derived from one donor whose serum gave equivocal CMV‐IgG results on FIAX. The sensitivity and specificity of the PLA assay were 100 percent for donor sera tested at both 4 and 22 degrees C and 91.5 and 98.4 percent, respectively, for platelet bag segment tests. Although no donors positive for CMV‐IgM were identified, 15 (14.4%) had equivocal IgM anti‐ CMV test results. The PLA ositivity of these samples correlated solely with the presence or absence of lgG as detected by FIAX. A total of 648 PLA tests were performed on donor sera and on platelet bag segments. On1 two false‐positive and five false‐negative results were observed. PLA testing for CMb antibody in donor sera and in ACPstored platelet concentrates is accurate and sensitive for up to 5 days. Segments may be remed from platelet bags for testing throughout the May shelf life of plateletpheresis components and Will Still provide accurate CMV testing result.
Donor room personnel attitudes toward autologous donors
Authors: Skettino S. , Ferguson K. , Andrews A. , Stehling L. C. , Strauss R. G. , Toy P. T.
Published: March 2003

Patients with serious illnesses who donate their blood for autologous use create anxiety for many phlebotomists. Donor room staff were surveyed at three donor centers, using confidential questionnaires, to identify their concerns and to evaluate factors associated with reduced anxiety toward autologous donors. Among respondents (n = 93), 58 percent had patient care experience within the last 5 years, which correlated with diminished concern about phlebotomizing cardiac patients (r = .21; p = 0.04). Fifty‐seven percent of respondents had drawn blood from greater than 50 autologous donors. Surprisingly, the experience of drawing blood from relatively large numbers of autologous donors did not correlate with increased confidence in phlebotomizing pediatric, cardiac, and elderly patients (r = ‐.04; p = 0.75). Those respondents who felt that they had adequate medical support agreed with more liberal donor criteria and were more confident about phlebotomizing pediatric, cardiac, and elderly patients (r = ‐.32; p = 0.001). Those respondents who believed most strongly in the benefits of autologous transfusion had the least anxiety toward donors who were medically more complex (r = .39; p = 0.0001). It is concluded that, contrary to what might be expected, phlebotomizing large numbers of autologous donors does not reduce the anxiety of staff members when they encounter donors with complex medical problems. Factors that do reduce anxiety include dependable physician and other medical support services, previous experience in direct patient care, and knowledge of the benefits of autologous transfusion.
Inhibition by albumin of merocyanine 540‐mediated photosensitization of platelets and viruses
Authors: Prodouz K. N. , Lytle C. D. , Keville E. A. , Budacz A. P. , Vargo S. , Fratantoni J. C.
Published: March 2003

The effect of the photosensitizer merocyanine 540 (MC 540) on platelets and on three marker viruses was examined to assess its potential in reducing virus transmission by blood products. The results demonstrated several deleterious effects of MC 540 (4–24 micrograms/mL) on platelet morphology and function in both the absence and presence of light (450‐ 600 nm). Treatment of washed platelets with MC 540 in the dark resulted in a significant release of serotonin in the absence of added agonist, as well as a diminished response to thrombin as measured in vitro. In addition, photosensitization caused spontaneous platelet aggregation and release of 92 percent of the releasable serotonin without the addition of an agonist. Because photo‐treatment of blood products is likely to be performed in a protein‐rich medium, the influence of albumin on the phototoxic effects on platelets was assessed. Albumin added to the suspension medium at concentrations greater than or equal to 1.0 percent protected the platelets against the effects of MC 540 in the dark, whereas 5‐percent albumin was required for protection against the phototoxic effects of MC 540 on the platelet response to thrombin. The antiviral activity of MC 540 and light was examined by using the lipid‐containing viruses herpes simplex virus (HSV) and bacteriophages phi 6 and PM2. Of the lipid‐enveloped viruses, HSV was 25 times more photosensitive to MC 540 than was phi 6 (15 micrograms/mL). PM2, which has an internal lipid layer, was almost 300 times less sensitive to MC 540 and light than was HSV. Photoinactivation of φ6 and PM2 (by 18 J/cm2) was completely prevented by the addition of 5‐percent albumin to the medium. Under these conditions, HSV was still inactivated but was nearly 200 times less sensitive. Therefore, the addition of albumin to protect platelets from damage induced by treatment with MC 540 and light reduced markedly the inactivation or viruses by such treatment.
An autologous blood program coordinated by a regional blood center: a 5‐ year experience
Authors: Giordano GF , Dockery J , Wallace BA , Donohoe KM , Rivers SL , Bass LJ , Fretwell RL , Huestis DW , Sandler SG
Published: March 2003

The Southern Arizona Regional Red Cross blood program offers preoperative autologous blood deposit to all patients and intraoperative autotransfusion services to all hospitals in the region. During a 5‐year period, the amount of preoperatively deposited autologous blood and intraoperatively salvaged red cells available increased from 0.3 to 19.6 percent of the community's total collections. Further increases in the availability and use of autologous blood may be achieved by community‐wide integration of services.
The impact of a patient education program on directed donations
Authors: Silver H , Lachman M , Badon S
Published: March 2003

The effect of an informational program in which the benefits and disadvantages of directed donations are discussed directly with the prospective recipient of the blood and/or that patient's family was examined during a 20‐month period. Data collected for 27 regional Connecticut hospitals that accept directed donations were compared with similar data for Hartford Hospital, an 885‐bed tertiary‐care facility. The number of directed‐donor units (68) collected by the American Red Cross Blood Services for Hartford Hospital during the study period was comparable to the number (average, 62.3) provided for smaller (301–450 beds) institutions in the state. The percentage that directed‐donor units drawn for Hartford Hospital represented of the total number of homologous units provided (0.23%) was less than that for any of the categories of hospitals, by size, in the state. Supplying the patient and/or the patient's family, in a personalized manner, with information concerning the advantages and disadvantages of directed as well as of volunteer donor blood can result in a marked overall reduction in the number of directed donations and also serves to reassure those concerned about blood transfusion.
Platelet crossmatching with lymphocytotoxicity test: an effective method in alloimmunized Chinese patients
Authors: Chow M.P. , Yung C.H. , Hu H.Y. , Tzeng J.L. , Lin W.M. , Lin C.K.
Published: March 2003

Fifty‐three patients receiving long‐term platelet transfusions were regularly screened for platelet‐associated antibodies by a platelet suspension immunofluorescence test (PSIFT) and a lymphocytotoxicity test (LCT). Subsequently, 24 patients became alloimmunized; all of their antibodies were of HLA specificity. Eighty‐two single‐donor platelet transfusions were given, and the clinical responses were considered satisfactory if the 18‐hour corrected count increment was 7.5 × 103 per μL or higher. In the meantime, 82 pairs of patient sera and donor lymphocytes were crossmatched. Among 63 crossmatched transfusions, 53 (84%) resulted in a satisfactory increment, with a mean (± SEM) of 17.71 ± 1.96 (× 103/μL), and 10 did not result in a satisfactory increment. The increments after 19 unmatched transfusions and 25 random‐donor (uncrossmatched) transfusions were 0.7 ± 0.3 and 2.39 ± 0.66, respectively. The difference was not significant (p > 0.05). The agreement between the LCT results and clinical response was 88 percent. Retrospectively, the corrected count increments showed no significant differences (p greater than 0.05) among three groups of HLA grading: the increments for A/BU/BX, C/D, and random HLA matches were 22.97 ± 4.07, 15.1 ± 1.97, and 14.85 ± 2.04, respectively. These results suggest that platelet crossmatching by LCT is an effective method for use in alloimmunized patients, especially Chinese patients.
Detection of anti‐A in neonatal serum
Authors: Mintz P.D. , Anderson G.
Published: March 2003

The thirteenth edition of the standards of the American Association of Blood Banks did not require the use of A1 red cells (RBCs) or an indirect antiglobulin test (IAT) to detect anti‐A in neonatal serum, whereas the fourteenth edition mandates both. The present study was conducted to help document the need for these changes. Incomplete expression of the A antigen in neonatal patients can contribute to the accumulation of unabsorbed maternal anti‐A that is capable of mediating the immune destruction of transfused adult RBCs. Sera from 50 group A neonatal infants of group O mothers were tested for anti‐A by using RBC segments of A1 and A2 units of AS‐1 RBCs less than 14 days old and also with A1 reagent RBCs. Of 22 sera in which anti‐A was detected by RBCs from the A1 units with an IAT, anti‐A was detected by RBCs from the A2 unit in only 1 and by the reagent RBCs in 17. In 19 (86%) of the 22 neonatal patients in whose sera anti‐A was detected, the antibody was found only with the use of anti‐human globulin in the IAT. It is concluded that testing to detect circulating anti‐A in neonatal patients should include an IAT, and that it is preferable to use A1 RBCs for the initial evaluation.
Rh33 in two of three German siblings with D+ C+ c+ E− e+ red cells
Authors: Moores P. , Smart E. , Sternberger J. , Schneider W.
Published: March 2003

The proposita in a German family of three siblings has D+ C+ c+ E‐ e+ f+ Rh: −17,19,33,34 red cells with weak C, e, f, Rh19, and Rh34 and stronger‐than‐usual Rh33 expression. One sibling has D+ C+ c+ E− e+ f+ Rh:17,19,33,34 red cells with weak f and ordinary‐strength Rh33, and the other sibling has D+ C+ c+ E− e+ f+ Rh:17,19,−33,34 red cells. In the absence of any further family members, the proposita's unusual phenotype suggests that she has an RoHar haplotype and a “new” Rh haplotype, provisionally named R1Lisa, that encodes Rh33, normal‐ strength D, weak C, weak or nondemonstrable e, Rh19, and Rh34, but not Rh17. Her Rh:33 sibling may have R1 and RoHar and her Rh:−33 sibling R1 and r haplotypes.
Effects of serial plasmapheresis on serum IgA levels in IgA‐deficient blood donors with IgA‐suppressor T cells
Authors: Paglieroni T.G. , Holland P.V.
Published: March 2003

Seventeen IgA‐deficient blood donors, without antibodies to IgA, underwent plasmapheresis four to eight consecutive times at intervals of 8 weeks or less to provide fresh‐frozen plasma for patients with anti‐IgA. Blood samples, drawn for analysis no more than 1 hour before plasmapheresis and again at the conclusion of each procedure, were analyzed for lymphocyte subpopulations and serum IgA levels. Five lymphocyte subpopulations, including natural killer cells, the suppressor‐inducer CD4 subset, the suppressor‐precursor CD8 subset, non‐major histocompatibility complex (MHC)‐restricted cytotoxic T cells, and CD5+ B cells, were all decreased significantly after plasmapheresis (p<0.05). In a subgroup of IgA‐deficient donors with excessive IgA‐suppressor T‐cell activity, serum IgA increased to levels exceeding 0.05 g per L following the fourth consecutive plasmapheresis procedure. Serum IgA levels did not similarly increase in IgA‐deficient donors without excessive IgA‐suppressor T‐cell activity or in controls without IgA deficiency. Our study shows the potential, in a subpopulation of IgA‐deficient donors who undergo frequent plasmapheresis, for a transient increase in serum IgA to a level no longer considered IgA deficient.
Analysis of the structure and activity of A and A,B immunoglobulin A monoclonal antibodies
Authors: Guest A.R. , Scott M.L. , Smythe J. , Judson P.A.
Published: March 2003

A study of the serologic activity and molecular structures of three spleen‐derived mouse IgA monoclonal human blood group‐specific supernatants was undertaken; this was part of an evaluation of these monoclonals as blood typing reagents. The monoclonal antibodies were eluted through a precalibrated size‐exclusion column, and fractions were analyzed by immunoblotting, heavy and light chain‐specific enzyme‐ linked immunosorbent assay, and liquid‐ and solid‐phase serologic tests. Results indicated that one of the supernatants (anti‐A specificity) contained tetrameric and monomeric forms of IgA, while the other two (anti‐A,B specificity) contained three higher polymeric forms (1000‐4000 kDa) and one dimeric form. The tetrameric and polymeric forms showed red cell agglutinating activity, whereas the dimeric and monomeric forms did not. All forms contained heavy and light chains. The monomeric anti‐A showed specific binding to appropriate red cells in a solid‐phase assay, but the dimeric anti‐A,B fractions did not. Purified fractions stored at 4 degrees C did not show any equilibration toward other forms, which indicated that the molecules are stable once secreted. The use of such antibodies as blood grouping reagents requires careful monitoring to ensure that high proportions of nonagglutinating molecular weight forms are not produced, as they could compromise the performance of the reagent by binding to red cell antigen in competition with the agglutinating forms.
A non‐computer‐dependent prospective review of blood and blood component utilization
Authors: Silver H. , Tahhan H.R. , Anderson J. , Lachman M.
Published: March 2003

The effect that a prospective review of blood and blood component utilization had during a 1‐year period at an 850‐bed tertiary‐care teaching hospital is reported. The review process, which has been in operation for 18 years, is based on institutionally developed usage guidelines for blood, fresh‐frozen plasma (FFP), platelets (Plts), and cryoprecipitate (Cryo). The guidelines were developed by the hospital's Transfusion Medicine Committee. The review is initiated by the transfusion service technical personnel when an order is received. Back‐ up support is provided, as needed, by physicians on the service or by those pathology residents taking off‐hour call. They consult with the ordering physician when questions arise as to the appropriateness of an order as it relates to the patient's clinical situation. Screening values used in the initial evaluation of each order are: hematocrit, less than or equal to 27 percent (0.27) for blood; prothrombin time, greater than or equal to 16 seconds, and/or activated partial thromboplastin time, greater than or equal to 60 seconds, for FFP; bleeding time, greater than or equal to 16 minutes and/or a platelet count of 20 to 100 × 10(3) per mm3 (20‐100 × 10(9)/L), as related to clinical conditions, for Pits; and fibrinogen less than or equal to 120 mg per dL (1.2 g/L) for Cryo. Also taken into consideration is the stated clinical indication, which is provided when the requisition is submitted. As a result of the prospective review, orders for blood and/or components for 114 patients were canceled, and 51 patients received 207 blood components that were more appropriate for the clinical situation. A total of 316 homologous donor exposures were avoided during the year surveyed. A total reduction in patient medical costs of $22,000 was realized as a result of canceled orders, a decrease in the number of units transfused, or the selection of a more appropriate component for transfusion.(ABSTRACT TRUNCATED AT 250 WORDS)
Pretransfusion incubation of apheresis platelets at 37°C improves posttransfusion recovery
Authors: SHANWELL A. , WIKMAN A. , RINGDBN O.
Published: March 2003

The effect of brief 37 degrees C pretransfusion incubation of apheresis platelets (n = 25) was studied in 14 allogenic bone marrow transplant recipients (5 children, 9 adults). Apheresis platelets were collected on a cell separator, divided in two aliquots, and stored 1 to 5 days at 22 degrees C during agitation. One aliquot was incubated at 37 degrees C for 1 hour before transfusion, and the other was transfused as a paired control. When the patient's peripheral platelet count fell below 30 × 10(9) per L, one aliquot was transfused. The other aliquot was transfused the following day. Each patient received the two aliquots in random order. Corrected count increment (CCI) at 1 hour after transfusion of 37 degrees C incubated platelets was 12.2 +/− 9.5 (mean +/− SD) compared to 7.5 +/− 5.4 for paired control platelets (p < 0.05). CCIs the next day were 4.4 +/− 6.1 and 2.4 +/− 2.7, respectively (not significant). It can be concluded that 37 degrees C incubation of apheresis platelets improves posttransfusion CCI.
Engraftment with peripheral blood stem cells collected by large‐volume leukapheresis for patients with lymphoma
Authors: COMENZO R.L. , MALACHOWSKI M.E. , MILLER K.B. , ERBAN J.J. , SCHENKEIN D.P. , DESFORGES J.F. , BERKMAN E.M.
Published: March 2003

Seven patients with refractory lymphomas underwent marrow reconstitution with peripheral blood stem cells (PBSCs) harvested by large‐volume leukapheresis (LVL). PBSCs were collected from all patients more than 1 month after the last cycle of chemotherapy, and no patient received growth factors. The median number of LVL procedures performed per patient was 4.5, with a mean volume of 24.5 L of blood processed per procedure to obtain 7 × 10(8) mononuclear cells per kg. Autologous PBSCs and platelets were frozen at a controlled rate in plasma and 10‐percent dimethyl sulfoxide and stored in the vapor phase of liquid nitrogen. This group of patients was compared to a control group (n = 18) who received medullary marrow (MM) transplants for the same diagnoses under the same protocols during the same period. Posttransplant days to white cell engraftment (PBSC = 17, MM = 15.5) were no different. Days to platelet independence were significantly longer in the LVL PBSC group (PBSC = 33, MM = 16; p < 0.05). This pattern of engraftment is typical of patients treated in this manner. Although Day 0 platelet counts (PBSC = 75.5 × 10(9)/L, MM = 85 × 10(9)/L) and total single‐donor unit platelet use (PBSC = 8, MM = 9) were no different, Day 1 platelet counts (PBSC = 128 × 10(9)/L, MM = 61.5 × 10(9)/L; p < 0.05) and Day 14 platelet use (PBSC = 5, MM = 8; p < 0.05) were significantly different, because of the transfusion of cryopreserved autologous platelets with PBSCs on Day 0.
Partitioning of hepatitis C virus during Cohn‐Oncley fractionation of plasma
Authors: YEI S. , Yu M.W. , TANKERSLEY D. L.
Published: March 2003

Because of concern about the safety of immune globulins with respect to transmission of hepatitis C, the partitioning of hepatitis C virus (HCV) during alcohol fractionation of a plasma pool prepared exclusively from anti‐HCV‐reactive donations was examined. Quantitation of HCV RNA was accomplished by nested polymerase chain reaction (PCR) at limiting dilutions. One PCR unit was arbitrarily defined as the minimum amount of HCV RNA from which an amplified product could be detected. The starting plasma pool contained 1.4 × 10(5) PCR units per mL. Most of the HCV RNA was found in cryoprecipitate and in Cohn fractions I and III, but it was also detected in fraction II, which is used for immunoglobulin G preparations. A 3.4‐percent solution of IgG prepared from this fraction II contained 30 PCR units per mL. The fractionation process leading to immune globulin resulted in overall reduction in HCV RNA by a factor of 4.7 × 10(4). Although the presence of HCV RNA in the final product does not necessarily imply the presence of infectious virus, this work suggests that the safety of immune globulins with respect to HCV transmission is not due solely to the partitioning of HCV away from the immunoglobulin fraction.
Recombinant immunoblot and polymerase chain reaction testing in volunteer whole blood donors screened by a multi‐antigen assay for hepatitis C virus antibodies
Authors: Sayers M.H. , Gretch D.R.
Published: March 2003

The purpose of this study was to compare the results of supplementary testing of volunteer whole blood donors who had been screened by the first hepatitis C virus antibody assay licensed in the United States with results from donors screened by a newer, more sensitive, multi‐ antigen assay. In contrast to the earlier assay, the multi‐antigen assay incorporates a recombinant hepatitis C virus antigen, c22‐3, which is encoded by a structural region of the viral genome. Supplementary testing included a second‐generation recombinant immunoblot assay and a highly sensitive polymerase chain reaction assay for evidence of hepatitis C virus genomic RNA. A comparison of supplementary test results reveals a higher percentage of donors screened by the newer assay to be indeterminate on recombinant immunoblot (34.4% vs. 6.4%, p < 0.05). Furthermore, polymerase chain reaction testing of donors with indeterminate blot results shows that 14 percent have evidence of viral RNA. For this reason, counseling of donors with indeterminate patterns on immunoblot must include informing them of the possibility that they are infected.
A comparison of polymerase chain reaction and an infectivity assay for human immunodeficiency virus type 1 titration during virus inactivation of blood components
Authors: Hart H. , McOmish F. , Hart W.G. , Simmonds P. , Yap P.L.
Published: March 2003

Three examples of human plasma‐derived concentrates, intermediate‐ purity factors VIII and IX, and fibrinogen were spiked with tissue culture‐grown human immunodeficiency virus type 1 (HIV‐1) strain RF. All examples were freeze‐dried and heated at 80 degrees C for 72 hours by using validated production process models. HIV‐1 infectivity was measured by a syncytial infectivity assay in C8166 cells and then compared with levels determined by nested HIV polymerase chain reaction (PCR). The infectivity assay demonstrated a reduction index of at least 4.5 log10, while PCR showed an average 1.7 log10. Large amounts of HIV‐ 1 RNA (105) were still detectable by PCR in samples in which infectivity assays failed to detect any HIV‐1. These data suggest that HIV‐1 PCR levels do not parallel HIV‐1 infectivity levels during virus‐ inactivation procedures involved in coagulation factor concentrate production. PCR was able to detect the RNA associated with inactivated HIV‐1 particles in the factor concentrates, which allows the conclusion that PCR is not a useful test with which to monitor virus‐inactivation procedures such as heating at 80 degrees C for 72 hours. This judgment contrasts with the more definite and sensitive role of PCR in diagnosing HIV‐1 infection in patients in whom a positive HIV‐1 PCR result correlates with active HIV‐1 infection and with PCR's usefulness in monitoring virus removal.
Production of modified crosslinked cell‐free hemoglobin for human use: the role of quantitative determination of endotoxin contamination
Authors: Roth R.I. , Levin J. , Chapman K.W. , Schmeizl M. , Rickles F.R.
Published: March 2003

In vivo toxicity remains a major barrier to the successful use of cell‐ free hemoglobin (Hb) as an oxygen carrier in humans. Bacterial endotoxin (lipopolysaccharide, LPS) is known to contribute to the in vivo toxicity of Hb preparations, and the prevention of LPS contamination is a critical aspect of the effort to create an efficacious Hb blood substitute. Limulus amebocyte lysate assays for endotoxin were performed on multiple Hb samples from 26 independent production runs for the preparation of human crosslinked cell‐free hemoglobin (alpha alpha Hb). High levels of LPS contamination (1‐ > 100 ng/mL) of alpha alpha Hb solutions were detected in multiple samples during many of the initial production runs. It was observed that LPS contamination of alpha alpha Hb solutions could occur at any step during the production sequence. Substantial enhancement by alpha alpha Hb of the biologic effects of LPS was demonstrated by two independent assays for endotoxin (the Limulus amebocyte lysate test and a mononuclear cell procoagulant assay), whereas LPS biologic activity was only slightly increased by human serum albumin and substantially diminished by IgG. These results suggest that the prevention of LPS‐ related toxicities in vivo may be more important to the clinical use of Hb solutions than to the use of other intravenous protein products. Therefore, it was encouraging to note that, with the careful monitoring for LPS in the production facility and in multiple samples during cell‐ free Hb production, sources of LPS contamination were recognized and the appropriate sites were made endotoxin‐free. Numerous subsequent production runs were performed without appreciable LPS contamination. Therefore, careful monitorinq of LPS contamination allows for the roduction of Hb solutions that are sufficiently free of LPS for clinical use.
White cell‐reduced red cells prepared by filtration: a critical evaluation of current filters and methods for counting residual white cells
Authors: Rebulla P. , Porretti L. , Bertolini F. , Marangoni F. , Prati D. , Smacchia C. , Pappalettera M. , Parravicini A. , Sirchia G.
Published: March 2003

White cell (WBC) reduction, red cell (RBC) recovery, and filtration time were determined in 1‐day‐old standard and buffy coat‐depleted RBCs filtered in the laboratory through six commercial filters for WBC reduction. Residual WBCs were counted with a Burker chamber (BC), with a Nageotte chamber (NC), and by flow cytometry (FC). Results show that BC counts were 0 in several cases in which WBCs were detected with NC and FC, which indicated that the traditional BC method is too insensitive in use with currently available filters. Calibration curves performed by FC and with NC with samples containing known concentrations of WBCs from 1000 to 1 per microL showed that both FC and NC detected, on average, 67 percent of WBCs present in the samples (efficiency). However, the efficiency of FC showed small variability (61–70%) at different WBC levels, whereas the variability with NC was large (39–91%). This greater variability prevented the correction of NC counts by using a single factor and indicated difficulty in NC standardization. Therefore, because our main aim was to compare different filters rather than to define absolute levels of WBC contamination, uncorrected FC and NC counts were chosen to be reported. True WBC counts per unit should not exceed values that can be obtained by dividing uncorrected counts by the lowest efficiencies (61% for FC and 39% for NC). Uncorrected NC and FC counts were below 2 × 106 per unit in all units processed through three of the filters and below 5 × 106 per unit in all units processed through the other three. Removal of the buffy coat from RBCs before filtration generally but not invariably caused slightly higher WBC removals. Median RBC recovery in CPD RBCs containing the buffy coat, filtered through four of the filters, was greater than 90 percent, which was significantly higher than that shown by the other two filters. Buffy coat removal prior to filtration involved an additional median loss of 12.4 percent of RBCs. Median filtration time ranged from 5 to 23 minutes per unit. Although NC appears to be less expensive than FC, its performance in experimental and routine conditions requires further evaluation and thorough standardization. Because of the continuous development of more effective filters, more sensitive methods for WBC counting in WBC‐reduced blood components are needed.
Treatment of refractoriness to platelet transfusion by protein A column therapy
Authors: Christie D.J. , Howe R.B. , Lennon S.S. , Sauro S.C.
Published: March 2003

Ten thrombocytopenic patients (platelets < 10–24 × 10(9)/L) who were refractory to platelet transfusion were investigated for their responsiveness to staphylococcal protein A column therapy. Nine patients had previously been treated with steroids, intravenous immune globulin, and/or other forms of immunosuppressive therapy without improvement in their transfusion response. All patients were receiving multiple platelet transfusions without achieving 1‐hour corrected count increments (CCIs) > or = 7500. Eight patients had antibodies that reacted with platelets and were directed against HLA class I antigens, ABO antigens, and/or platelet‐specific alloantigens. Plasma (500–2000 mL) from each patient was passed over a protein A silica gel column and then returned to the patient. Patients received from 1 to 14 treatments. A positive response to protein A therapy was defined as at least a doubling of the pretreatment platelet count and/or two successive 10‐ to 120‐minute posttransfusion CCIs > or = 7500. Following plasma treatments, 6 of 10 patients responded with daily platelet counts that averaged 48 ±/− 11 × 10(9) per L as compared with counts of 16 ±/− 7 × 10(9) per L (p < 0.0005) before treatment. Posttransfusion CCI values determined in four of these patients averaged 2480 ±/− 810 and 10,010 ±/− 3540 (p < 0.005) before and after treatment, respectively. In contrast, among the four unresponsive patients, platelet counts averaged 10 ±/− 9 and 13 ±/− 10 × 10(9) per L (p = NS), respectively, while posttransfusion CCIs were 700 ±/− 1410 and 1520 ±/− 2460 (p = NS), respectively. while osttransfusion CCls were 700 2 1410 and 1520 2 2460 (p = NS), respectively. These findings suggest that, in certain patients who are refractory to platelet trans‐ fusion because of platelet‐reactive antibodies, protein A column treatments may be an effective means of increasing platelet counts and responsiveness to platelet trans‐ fusions.
Characteristics of red cells irradiated and subsequently frozen for long‐term storage
Authors: Suda B.A. , Leitman S.F. , Davey R.J.
Published: March 2003

Irradiation of blood components eliminates the risk of transfusion‐ associated graft‐versus‐host disease. Freezing directed or rare red cell units that are irradiated but not transfused would facilitate inventory management and would increase the transfusion options for the involved patients. However, no studies have been performed to evaluate whether prestorage irradiation damages subsequently frozen red cells. Ten normal volunteers donated a unit of whole blood on two separate occasions. One unit was irradiated with 15 Gy (1500 rad), stored at 4 degrees C for 6 days, and then frozen and stored at ‐75 degrees C for 56 days. The other unit (control) was similarly stored but was not irradiated. Aliquots of the units were tested on Day 0 and Day 6 and, after deglycerolization, on Day 62. Comparison of means and changes in means showed no significant differences in red cell ATP, 2,3 DPG, or supernatant hemoglobin and glucose in control and irradiated units. The difference in the change in supernatant potassium from Day 0 to Day 6 in control and irradiated units was significant (1.5 to 28.6 mmol/L vs. 1.5 to 48.5 mmol/L: p < 0.0001). Irradiation did not cause significant differences in postdeglycerolization red cell recovery (control, 84.5% vs. irradiated, 81.2%) or in 24‐hour posttransfusion autologous red cell survival (control, 91.1% vs. irradiated, 90.9%). Red cells can be irradiated, stored at 4 degrees C for 6 days, and subsequently frozen with no increase in detectable damage as compared to controls that were not irradiated.
Stability of red cell antigens and plasma coagulation factors stored in a non‐diethylhexyl phthalate‐plasticized container
Authors: Snyder E.L. , Hedberg S.L. , Napychank P.A. , Roberts C. , Kagen L. , Aster R.A. , Quinlan K. , Strucaly A. , Buchholz D.H.
Published: March 2003

Red cell antigen stability studies were performed to evaluate whether the storage of red cells in plastic segments made up of a new non‐di‐2(ethylhexyl)phthalate (DEHP)‐plasticized material resulted in poststorage antigenic reactivity different from that seen in segments made from DEHP‐containing plastic. Serial 1‐in‐2 dilutions of commercially available antisera were prepared and tested by using stored red cells obtained from segments on Days 0, 28, 42, and, in some instances, 49. Antigenic determinants tested included A, B, D, c, K, Lea, Fya, Jka, M, and P1. To minimize variability, the same reagent lots were used throughout each study, and the same technologists performed the assays in each laboratory. No significant differences in titration scores were seen when cells stored in segments made of the test plastic were compared with cells obtained from the same donor and stored for the same length of time in segments made of control plastic. In addition, plasma coagulation factor stability was studied in fresh‐frozen plasma and cryoprecipitate stored for up to 1 year in the non‐DEHP‐plasticized plastic containers. No significant differences were seen in prothrombin time, activated partial thromboplastin time, fibrinogen content, or factor V, VII, VIII, IX, or × activity as compared with plasma stored for equal periods of time in control plastic containers. It is concluded that the test plastic does not adversely affect red cell antigenic reactivity or plasma coagulation factor stability and that it is suitable for use in clinical transfusion practice.
Granulocyte transfusions: efficacy in treating fungal infections in neutropenic patients following bone marrow transplantation
Authors: Bhatia S. , McCullough J. , Perry E.H. , Clay M. , Ramsay N.K. , Neglia J.P.
Published: March 2003

Background: A retrospective study was conducted to evaluate the efficacy of granulocyte transfusions in neutropenic patients with fungal infections following bone marrow transplantation.

Study Design and Methods: Systemic fungal infection was detected in 87 patients during the first 100 days following bone marrow transplantation; 50 received granulocytes in addition to appropriate antifungal agents. The median age was 17 years in the transfused patients (range, 1.5–57) and 35 years in the nontransfused patients (range, 0.8–50). Granulocyte transfusions were given on a daily to twice‐daily basis. To evaluate their responses, patients were categorized by infection type (candidal [n = 38] vs. noncandidal [n = 49]) and site (fungemia alone [n = 30] vs. invasive infection [n = 57]). Resolution of infection was defined as the resolution of signs and symptoms and negative cultures and/or histopathology.

Results: No benefit of granulocyte transfusions could be shown in the resolution of infection in patients with either invasive noncandidal infection (29% in the transfused patients vs. 23% in the nontransfused patients, p > 0.1) or candidal sepsis (56% vs. 50%, p > 0.1). Among patients with delayed marrow recovery, no difference was seen in the resolution of infection in the transfused (25.9%) and nontransfused (50%) patients (p > 0.1); nor was any difference between the transfused and nontransfused patients evident in the duration of febrile episode associated with the fungal infection. Granulocyte transfusions were well tolerated, with the only complications being fever in 12 patients (24%), chills in 10 (20%), and respiratory distress in 2 (4%). Despite attempts to stratify by infection type, invasiveness, and marrow recovery, it was not possible to show any benefit of granulocyte transfusions in this group.

Conclusions: It is likely that only through a prospective randomized trial can the question of the efficacy of granulocyte transfusions in treating fungal infections be conclusively answered.
Fatal disseminated intravascular coagulation and pulmonary thrombosis following blood transfusion in a patient with severe autoimmune hemolytic anemia and human immunodeficiency virus infection
Authors: Bilgrami S. , Cable R. , Pisciotto P. , Rowland F. , Greenberg B.
Published: March 2003

Background: Autoimmune hemolytic anemia (AIHA) has rarely been reported in association with human immunodeficiency (HIV) infection and never as a presenting manifestation. Similarly, disseminated intravascular coagulation (DIC) is a very infrequent complication of HIV infection.

Case Report: An unusual patient is described who at the time of presentation with severe AIHA was found to be HIV positive. Shortly thereafter, he developed DIC, pulmonary thrombi, and right heart failure that proved fatal, in spite of intensive supportive measures.

Conclusion: Although the etiology of the DIC and pulmonary thrombi could not be established, they are most likely related to aggressive transfusion therapy, with associated intravascular hemolysis.
Inactivation of lipid‐enveloped and non‐lipid‐enveloped model viruses in normal human plasma by crosslinked starch‐iodine
Authors: Highsmith F.A. , Xue H. , Caple M. , Walthall B. , Drohan W.N. , Shanbrom E.
Published: March 2003

Background: The purpose of this study was to evaluate an iodine‐based method of activating potentially harmful viruses that might be found in normal human plasma.

Study Design and Methods: A procedure has been developed for inactivating lipid‐enveloped and non‐lipid‐enveloped model viruses in normal human plasma by using iodine complexed to crosslinked potato starch. The established conditions are an iodine concentration of 1.05 mg per mL and 60 minutes' incubation.

Results: Under these conditions, inactivation of more than 9 log10 of vesicular stomatitis virus, a lipid‐enveloped virus, and more than 7 log10 of encephalomyocarditis virus, a non‐lipid‐enveloped virus, was achieved, with minimal losses of biologic activity of selected plasma proteins. Under these conditions, 70 percent of factor VIII activity, 77 percent of factor IX activity, and 100 percent of protein C activity in the plasma were retained.

Conclusion: Crosslinked starch‐iodine may be useful in the inactivation of viruses in single‐donor plasma units and in pooled human plasma before fractionation.
Blood collections by community blood centers, 1988 through 1992
Authors: Forbes J.M. , Laurie M.L.
Published: March 2003

BACKGROUND: The provision of a safe and sufficient supply of blood is critical to patient care.

STUDY DESIGN AND METHODS: A survey was conducted of the blood collection activity of the 189 community blood centers operating in the United States from 1988 through 1992. Data were analyzed by source of the donation (allogeneic or autologous), by center's collection volume, and by geographic region. Total collection figures were compared to historical blood collection activity since 1970.

RESULTS: A total of 12.31 million units of blood were collected in 1992, an increase of 2.6 percent over the total number of units collected in 1988. For the 5‐year period (1988–1992), total collections increased at a compound annual growth rate of 0.6 percent. The collection of allogeneic blood units declined by 0.2 percent annually, while that of autologous units increased by 23.2 percent annually. Autologous blood units accounted for 5.7 percent of total collections in 1992. Nationally, 48 units were collected per 1000 people in 1992, although substantial geographic variation (range, 38–64 units/1000) was observed across nine US census regions.

CONCLUSION: The data from this study provide evidence that the total supply of blood grew more slowly from 1988 through 1992 than in the years before 1988.
Guidelines for blood utilization review
Authors: Stehling L. , Luban N.L. , Anderson K.C. , Sayers M.H. , Long A. , Attar S. , Leitman S.F. , Gould S.A. , Kruskall M.S. , Goodnough L.T. , Hines D. M.
Published: March 2003

Hospitals are required by accrediting agencies to perform blood utilization review. Specific areas that must be addressed are the ordering, distribution, handling, dispensing, and administration of blood components. Monitoring the effects of transfusion on patients is also required. The format of the review process and the criteria for appropriate blood utilization must be developed by each institution. This article provides examples of areas that can be reviewed and procedures that may be used. However, the suggested laboratory values must not be interpreted as defining indications or criteria for transfusion. Each transfusion committee, or its equivalent, is responsible for developing its own institutional blood utilization procedures and audit criteria. Review and approval by the medical staff prior to implementation are essential. The procedures must also be reviewed and revised on a regular basis.
Abnormal neutrophil phenotype and neutrophil FcRIII deficiency corrected by bone marrow transplantation
Authors: Minchinton R.M. , de Haas M. , von dem Borne AE , Kleijer M. , Roberts A.W. , Gillett E.A.
Published: March 2003

BACKGROUND: Neutrophils from a patient in first remission of acute myeloid leukemia were found to lack NA1 and NA2 alloantigens. This NA null phenotype was converted to the normal phenotype of NA1, NB2 by the transplantation of bone marrow from an HLA‐identical sibling. To investigate the inherited or acquired nature of this rare phenotype, a combination of conventional neutrophil serology and recently developed restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR) assays was used.

STUDY DESIGN AND METHODS: Diagnosis, remission, and posttransplant patient peripheral blood samples were used for neutrophil phenotyping by granulocyte agglutination and immunofluorescence tests. The presence and dose of the gene for neutrophil Fc gamma RIIIb (Fc gamma RIIIB) were tested for with RFLP and Southern analysis and PCR‐based RFLP tests. Plasma levels of circulating soluble Fc gamma RIII (sFc gamma RIII) were measured with radioimmunoassay. The sibling bone marrow donor and the patient's parents were also studied.

RESULTS: RFLP analysis of DNA obtained from the patient at the time of diagnosis showed that she lacked the Fc gamma RIIIB gene for neutrophil Fc gamma RIII (i.e., Fc gamma RIIIb), but that, in DNA prepared from posttransplant samples, the Fc gamma RIIIB gene was present. Quantitation of plasma levels of soluble FcRIII (sFcRIII) demonstrated a complete absence of sFcRIII in the patient's pretransplant plasma. However, 20 units of sFcRIII were detected in the patient's plasma by 160 days after graft. Hair samples from the patient provided sufficient nonhematopoietic, genomic DNA to confirm that her genotype was NA0NA0. DNA prepared from lymphocytes of both parents and the sibling marrow donor was used to quantitate their Fc gamma RIIIB gene dose. The mother and brother had only one Fc gamma RIIIB gene each, while the father apparently had a normal complement of two Fc gamma RIIIB genes.

CONCLUSION: In this case, an inherited absence of Fc gamma RIIIB gene in a patient with acute myeloid leukemia was unintentionally corrected by the transplantation of bone marrow from a sibling donor who himself carried only one Fc gamma RIIIB gene.
A cost analysis comparing erythropoietin and red cell transfusions in the treatment of anemia of prematurity
Authors: Fain J. , Hilsenrath P. , Widness J.A. , Strauss R.G. , Mutnick A.H.
Published: March 2003

BACKGROUND: Anemia of prematurity is invariably observed in very low birth weight infants and may become symptomatic enough to be treated with packed red cell transfusions. Recently, treatment of this condition with recombinant human erythropoietin has been advocated.

STUDY DESIGN AND METHODS: To compare the costs of training symptomatic anemia in hospitalized premature infants with transfusions alone or with erythropoietin plus red cell transfusions as needed, cost estimates were derived from local hospital and published cost data. Decision analysis and sensitivity analysis were applied to a “base case.” The base case was derived from results of a multicenter erythropoietin trial in the United States in which premature infants received 500 U of erythropoietin per kg of body weight each week. Because erythropoietin treatment began on average at 3 weeks of life, when infants were clinically stable, they had already received 3.5 red cell transfusions. During the 6‐week treatment period, erythropoietin‐ treated infants received significantly fewer additional transfusions: a mean of 1.6 versus 1.1.

RESULTS: The base‐case cost in 1993 dollars for treating anemia in hospitalized premature infants with erythropoietin and transfusions was $1,326. This was nearly twice the cost of conventional treatment with transfusions alone ($721). If the 6‐week treatment period alone is considered, erythropoietin is 3.6 times more costly: $840 versus $235.

CONCLUSION: The largest available US study using erythropoietin to treat anemia in premature infants has demonstrated a small, but significant, reduction in transfusion needs. However, this study's cost data alone do not justify the widespread use of erythropoietin in premature infants. When this issue is probed in great depth, sensitivity analyses demonstrate that major reductions in erythropoietin's cost and/or improvements in its effectiveness quite possibly will make its use economically more attractive.