bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti‐Jka, ‐C, and ‐E in a single patient, initially demonstrable only by the manual hexadimethrine bromide (Polybrene) test, with incompatibilities confirmed by 51Cr‐labeled red cell studies
Authors: Maynard B.A. , Smith D.S. , Farrar R.P. , Kraetsch R.E. , Chaplin H.
Published: March 2003

Published reports have confirmed the superior sensitivity of the manual hexadimethrine bromide (Polybrene) test (MPT) for demonstrating many alloantibodies in vitro; however, the clinical significance of alloantibodies demonstrable exclusively by MPT has not been shown conclusively. A patient with macroglobulinemia experienced chills, fever, hemoglobinemia, and hemoglobinuria following the transfusion of 1 unit of red cells (RBCs) shown to be compatible by the low‐ionic‐strength antiglobulin (LIS‐AG) method. Serologic investigation was negative. Intravascular hemolysis occurred with a second “compatible” unit. Serologic studies were again negative by LIS‐AG and ficin‐AG methods, but revealed anti‐Jka by MPT. Both donors were Jk(a+b−), and 51Cr studies of the second donor's RBCs revealed a t½ of < 30 minutes, with marked intravascular hemolysis. A LIS‐AG‐compatible Jk(a−) unit was transfused uneventfully, but with no rise in hematocrit. MPT next revealed anti‐C; subsequent 51Cr studies with the Jk(a−), Cc donor's RBCs showed a 51Cr t½ of 100 minutes with slight intravascular lysis. Four transfusions of Jk(a−), C− blood were uneventful, but 5 days later the patient's hemoglobin declined. The following day, anti‐E was demonstrable exclusively by MPT. 51Cr‐labeled Jk(a−), C−, E− RBCs had normal 24‐hour survival. The patient's hemoglobin rose to 11 g per dl following transfusions of Jk(a−), C−, E− RBCs, and he was discharged. In vitro studies employing the patient's purified IgM paraprotein revealed no interference with alloantibody binding or detection.
False‐positive results with chemically modified anti‐D do not indicate a need to use a separate, immunologically inert Rh control reagent
Authors: Judd W.J. , Steiner E.A. , Oberman H.A. , Giacherio D.
Published: March 2003

Thirteen of 1450 (0.9%) Rh‐negative samples submitted for pretransfusion or prenatal testing gave weak false‐positive reactions in immediate‐spin (IS) tests with chemically modified anti‐D (CM‐D). Only seven reacted in control tests with anti‐A and/or ‐B. The other six samples could have been considered Rh positive; however, such a conclusion is inappropriate, since stronger (> 3+) reactions are expected in IS tests with CM‐D and the vast majority of true Rh‐positive blood samples. Moreover, three of the six false‐positive CM‐D reactions associated with negative control tests were from patients previously found to be Rh negative. The other three samples gave 1+ to 2+ reactions with CM‐D and did not react with anti‐A and/or ‐B; these erroneous CM‐D reactions were recognized by confirmatory tests with high‐protein anti‐D and Rh control reagents that were performed because previous typing results were not on file. Such confirmation, as well as careful grading and proper interpretation of serologic reactions, serves to prevent erroneous Rh typing without the use of a separate, immunologically inert Rh control reagent.
The distribution of blood group antigens and alloantibodies among Chinese in Taiwan
Authors: Lin‐Chu M. , Broadberry R.E. , Chang F.J.
Published: March 2003

Most blood group antigens among Chinese people are very homogeneous (e.g., D, 99.4%; K, 0%; Fya, 99.7%). The frequency of clinically significant alloantibodies detected by the authors' crossmatching was only 0.146 percent, suggesting that pretransfusion testing can be greatly simplified in Chinese populations.
Viability of platelets following storage in the irradiated state. A pair‐controlled study
Authors: Read E. J. , Kodis C. , Carter C. S. , Leitman S. F.
Published: March 2003

Gamma irradiation of blood products is a standard practice recommended for the prevention of posttransfusion graft‐versus‐host disease in susceptible hosts. We studied the effects of irradiation on stored platelet concentrates and evaluated whether platelets could be stored for 5 days in the irradiated state without adverse effects on their viability. Using a pair‐controlled design in which each of six normal subjects acted as his or her own control, we compared in vitro storage characteristics and in vivo kinetics of platelet concentrates exposed to 30 Gy and stored for 5 days with those of platelet concentrates simply stored for 5 days without irradiation. Irradiation had no significant effects on in vitro storage characteristics (platelet count, mean platelet volume, pH, and white cell count) or on in vivo kinetics, including initial recovery and mean platelet survival. Using the multiple‐hit model, initial recovery was 49.6 ± 10.8 percent, and mean platelet survival was 5.6 ± 1.05 days for irradiated concentrates, compared with 51.3 ± 13.0 percent and 5.9 ± 0.50 days, respectively, for the unirradiated control concentrates. We conclude that irradiation of platelet concentrates with up to 30 Gy has no effect on their in vivo recovery or survival, and that irradiation administered before storage of platelet concentrates does not interfere with their clinical efficacy.
Determination of zygosity using flow cytometric analysis of red cell antigen strength
Authors: Oien L. , Nance S. , Arndt P. , Garratty G.
Published: March 2003

A flow cytometry method, developed in our laboratory to measure red cell (RBC)‐bound IgG, was compared to the manual titration technique in the measurement of RBC antigen strength to determine zygosity. Parallel studies using antibodies to antigens in the Rh, Kell, Kidd, Ss, and Duffy systems were performed. The antisera (n = 20) were tested against five examples each of RBCs from apparent homozygotes and heterozygotes. The flow cytometry method was clearly superior, showing distinct differences, with no overlap of the ranges of results, between the reactions of RBCs from homozygotes and heterozygotes with 10 of 20 (50%) antisera. By the manual titration technique none of these sera clearly demonstrated dosage and 15 showed overlap of the ranges. It was obvious from the results that the commonly used manual titration technique for comparing the test RBCs with a single example of RBCs from a homozygote and heterozygote yielded inaccurate results.
A cost‐effectiveness evaluation of platelet crossmatching and HLA matching in the management of alloimmunized thrombocytopenic patients
Authors: Freedman J. , Gafni A. , Garvey M. B. , Blanchette V.
Published: March 2003

Effective platelet support for alloimmunized refractory thrombocytopenic patients may be provided by several potential strategies, the most common being HLA‐matched single‐donor platelets or crossmatch‐compatible, pooled random‐ or single‐donor platelets. This study used a detailed economic analysis to compare the cost‐effectiveness of several techniques for platelet crossmatching and that of HLA‐matched single‐donor platelets. The crossmatch methods evaluated were a microlymphocytotoxicity test (LCT), an immunofluorescence technique (PSIFT), a radioactive antiglobulin test (PRAT), and an enzyme‐linked immunosorbent assay (ELISA). The analysis was based on the need to support 100 refractory patients with acute leukemia with a presumed requirement of 500 transfusions. The relative costs for a successful crossmatch were: PRAT < LCT < LCT + PRAT < PSIFT < ELISA. In the comparison of the crossmatch methods, an increase in costs was generally associated with an increase in the number of successful transfusion episodes. However, decreasing marginal gains were seen. The HLA‐matched single‐donor platelets were relatively cost‐inefficient in comparison to the crossmatch‐compatible platelets. A theoretic sequence of tests for cost‐effective provision of optimal platelet support in refractory patients was evaluated. Such considerations of cost are important in the selection of an optimal program for the management of alloimmunized refractory thrombocytopenic patients.
Blood group phenotypes in Taiwan
Authors: Yung C. H. , Chow M. P. , Hu H. Y. , Mou L. L. , Lyou J. Y. , Lee T. D.
Published: March 2003

This report describes a study of 31 red cell antigens in 13 blood group systems tested over a period of 3 years in the Chinese population of Taiwan. The study provides evidence that major differences exist between Taiwanese and whites or blacks in five blood group systems: Rh, MNSs, Duffy, P, and Xg.
Fatal erythroderma (suspected graft‐versus‐host disease) after cholecystectomy. Retrospective analysis
Authors: Otsuka S. , Kunieda K. , Hirose M. , Takeuchi H. , Mizutani Y. , Nagaya M. , Sato G. , Kasuya S. , Matsutomo K. , Noma A. , Saji S.
Published: March 2003

Fatal postoperative erythroderma (POE) developed in a 52‐year‐old woman with gallstones who underwent elective cholecystectomy. During surgery, she was transfused with 3 units of unirradiated packed red cells stored in the liquid state for at least 4 days after collection. The POE is believed to have been the result of transfusion‐associated graft‐versus‐host disease (TA‐GVHD). The diagnosis of GVHD was based upon the characteristic clinical picture and retrospective HLA typing. The implicated donor was homozygous for an HLA haplotype that appeared to be shared with the recipient: A24‐CBL‐Bw52‐DR2‐DRw52‐DQw1, the most common haplotype in the Japanese population. This case raises the possibility that a transfusion of relatively fresh blood from a donor who has no HLA antigens incompatible with the recipient may result in GVHD in patients with no apparent immunoincompetence who are undergoing relatively minor surgery with no chemotherapy or radiation therapy.
Functional properties of a new crosslinked hemoglobin designed for use as a red cell substitute
Authors: Keipert P.E. , Adeniran A.J. , Kwong S. , Benesch R.E.
Published: March 2003

A new crosslinking agent, bis‐pyridoxal tetraphosphate, (bis‐PL)P4, was used to prevent dissociation of the hemoglobin (Hb) tetramer. Yields in excess of 75 percent of intramolecularly crosslinked (bis‐PL)P4Hb have been obtained using stoichiometric amounts of the crosslinking reagent. Some functional properties of (bis‐PL)P4Hb have been evaluated in vitro and in vivo. Oxygen affinity was substantially reduced (p50 = 31 torr at 37°C, pH 7.4, pCO2 = 40 torr), while the Bohr coefficient was −0.27 of H+ per mol of O2. Owing to its right‐shifted dissociation curve, (bis‐PL)P4Hb still yielded a p50 of 15 torr at a low temperature (16°C), as compared with only 3 torr for normal adult Hb (HbA). Clearance of (bis‐PL)P4Hb from plasma was significantly delayed (t½ = 171 min, at a dose of 0.2 g/kg of body weight compared with that of HbA (t½ = 54 min). Heart rate, mean arterial blood pressure, and respiration remained stable or returned to normal values within hours after bolus injection of the hemoglobin. The (bis‐PL)P4Hb was not excreted in the urine, in contrast to HbA (21% of the total dose of HbA appeared in the urine within the first 2 hrs). These results show that the covalent β‐β crosslink prevents the renal excretion of (bis‐PL)P4Hb, thereby significantly prolonging vascular retention. These properties, together with an increased ability to unload O2, make (bis‐PL)P4Hb a promising new candidate as a red cell substitute.
Red cell transfusion therapy for anemia in patients with AIDS and ARC: incidence, associated factors, and outcome
Authors: Jacobson M.A. , Peiperl L. , Volberding P.A. , Porteous D. , Toy P.T.C.Y. , Feigal D.
Published: March 2003

The records of the San Francisco General Hospital (SFGH) Blood Bank were reviewed, and 263 likely AIDS and AIDS‐related complex (ARC) patients were identified, who received 1545 units of packed red cells (PBRCs) between July 1, 1987, and June 30, 1988. A probability sample of 80 of these patients was selected randomly for detailed chart review. Of this sample, 78 (98%) were confirmed to have AIDS (86%) or ARC (14%). On the basis of the yearly census of the SFGH AIDS clinic, a transfusion incidence of 0.89 PRBC units per patient per year for patients with AIDS and 0.27 PRBC units per patient per year for those with ARC was estimated. Whereas 26 percent of the 177 transfusions studied in detail involved more than one associated (possibly causative) factor, antimicrobial drug therapy, zidovudine therapy, and disseminated Mycobacterium avium complex (MAC) infection were the sole associated factors in 20, 14, and 12 percent of the transfusions, respectively. To assess the role of MAC, the 263 transfused patients were compared with the 574 patients whose blood was submitted to the SFGH Mycobacteriology Laboratory during the same period. Patients whose blood yielded MAC had a relative risk of 5.2 for transfusion‐requiring anemia. In 80 percent of cases, the patient returned home after transfusion. Most PRBC transfusions administered to AIDS or ARC patients were optimal therapy.
Sepsis associated with transfusion of red cells contaminated with Yersinia enterocolitica
Authors: Tipple M.A. , Bland L.A. , Murphy J.J. , Arduino M.J. , Panlilio A.L. , Farmer J.J. , Tourault M.A. , Macpherson C.R. , Menitove J.E. , Grindon A.J. , Johnson P.S. , Strauss R.G. , Bufill J.A. , Ritch P.S. , Archer J.R. , Tablan O.C. , Jarvis W.R.
Published: March 2003

Between April 1987 and May 1989, the Centers for Disease Control investigated seven cases of transfusion‐associated Yersinia enterocolitica sepsis; four were caused by organisms of serotype O:3, and one each was caused by organisms of serotype O:1,2,3; O:5,27; and O:20. All seven recipients developed septic shock after receiving units of red cells (RBCs) contaminated with Y. enterocolitica; five recipients died. The cases occurred in seven states and were unrelated. There was no evidence for contamination of the RBC units during processing. Six of the seven donors had serologic evidence of recent Y. enterocolitica infection, and it is hypothesized that these donors had asymptomatic bacteremia when they donated the implicated blood. Four of the seven donors reported gastrointestinal illness in the 4 weeks before blood donation, and one donor became ill on the day he donated blood. Y. enterocolitica grows well at 4° C and in the presence of dextrose and iron. If blood is contaminated at the time of collection, storage of the RBCs at 4° C provides an ideal environment for bacterial growth and endotoxin production. These cases demonstrate the need for careful evaluation of patients with transfusion reactions for possible sepsis and suggest a need to screen prospective blood donors for mild gastrointestinal illness, including those illnesses not requiring physician evaluation or medication.
Evaluation of the polyethylene glycol‐potentiated indirect antiglobulin test
Authors: Wenz B. , Apuzzo J. , Shah D.P.
Published: March 2003

Results of the indirect antiglobulin test (IAT) potentiated by the addition of polyethylene glycol (PEG) were compared with those of the IAT performed with bovine serum albumin (BSA). This retrospective study, which encompasses 3 years, confirms that use of PEG‐IAT increases the rate of detection of clinically significant red cell (RBC) antibodies, decreases the detection of insignificant antibodies, reduces the need to use other serologic enhancement techniques (such as enzyme modification of RBCs), and has a positive impact on the laboratory's workload. These advantages are provided with no diminution in the quality of service provided by the blood bank.
A METHOD FOR COUNTING LEUKOCYTES IN FILTERED COMPONENTS
Authors: GREENWALT TJ , ALLEN CM
Published: March 2003

ABSTRACT: Leukocyte‐depleted components are being used more frequently. Quantitation of residual WBCs is needed for quality control and for establishing the critical level required to prevent alloimmunization. Electronic counting is not accurate at WBC levels below 100/μL (1) and previously proposed techniques are either tedious or require sophisticated equipment. We describe a simple hemacytometer procedure using Türks solution that gives 87–93% recovery at WBC levels below 40/μL.
Detection of drug‐dependent, platelet‐reactive antibodies by antigen‐ capture ELISA and flow cytometry
Authors: Visentin G. P. , Wolfmeyer K. W. , Newman P. J. , Aster R. H.
Published: March 2003

The effectiveness of flow cytometry in the detection of drug‐dependent, platelet‐reactive antibodies was investigated. In studies of seven sera known to contain quinine‐ or quinidine‐dependent, platelet‐reactive antibodies, flow cytometry was 5 to 10 times more sensitive in detecting drug‐dependent antibodies (DDAbs) than the 51Cr release assay, antigen‐capture enzyme‐linked immunosorbent assay (ELISA), and indirect immunofluorescence microscopic assay. With flow cytometry, DDAbs could be detected at drug concentrations as low as 0.1 microM, or less than one‐tenth the level required with other methods. Antigen‐ capture ELISA was not as sensitive as flow cytometry in DDAb detection, but it did allow identification of the DDAbs' target molecules. With this assay, five of the seven DDAbs recognized both the glycoprotein Ib/IX (GPIb/IX) and glycoprotein IIb/IIIa (GPIIb/IIIa) complexes, while the remaining two sera reacted only with GPIb/IX. Of 44 consecutive patients who developed thrombocytopenia while taking quinidine, DDAbs were detected by flow cytometry in 11 (25%), more than twice the number detected by other methods. In one patient who developed thrombocytopenia while taking trimethoprim/sulfamethoxazole, DDAbs could be detected only by flow cytometry. It can be concluded that flow cytometry is highly sensitive in detecting DDAbs and allows their detection at pharmacologic concentrations of the drug. Most quinidine‐ dependent antibodies recognize at least two different glycoprotein complexes in the platelet membrane.
Lack of efficacy of tranexamic acid in thrombocytopenic bleeding
Authors: Fricke W. , Alling D. , Kimball J. , Griffith P. , Klein H.
Published: March 2003

A controlled, randomized, double‐blind study was performed to assess the effect of the oral antifibrinolytic agent tranexamic acid in patients with amegakaryocytic thrombocytopenia as regards their need for platelet transfusions and the number of bleeding episodes experienced. Each patient served as his or her own control and received sequential, randomized courses of either tranexamic acid or an identical placebo. The need for platelet transfusions due to bleeding and the total number of bleeding episodes were compared for tranexamic acid and placebo courses. Patients received platelet transfusions at the discretion of their personal physician and kept detailed records of bleeding episodes. Of three patients who completed the full study, none had a reduction in the need for platelet transfusions. Moreover, in the eight patients who participated in the study, there was no reduction in number of bleeding episodes during tranexamic acid treatment as compared to the number with placebo. Our data indicate that the prophylactic administration of tranexamic acid does not decrease dependence on platelet transfusions or decrease bleeding episodes in patients with bleeding due to amegakaryocytic thrombocytopenia.
Improvement in transfusion safety using a new blood unit and patient identification system as part of safe transfusion practice
Authors: Wenz B. , Burns E. R.
Published: March 2003

A new patient and blood unit identification system designed to confirm the identity of crossmatched blood products and that of the intended recipient was evaluated. Six hundred seventy‐two red cell concentrates were transfused to 312 patients. Participating hospital personnel and patients were interviewed regarding the use and benefit of this unique system, which incorporates a “lock‐box” approach to the identification process. The product and procedure were accepted unanimously and enthusiastically, and three potential mistransfusions were avoided by use of the system during the limited period of observation. This type of approach to the identification process affords greater security than conventional practices and minimally burdens staff.
Screening potential blood donors at risk for human immunodeficiency virus
Authors: Mayo D. J. , Rose A. M. , Matchett S. E. , Hoppe P. A. , Solomon J. M. , McCurdy K. K.
Published: March 2003

ABSTRACT: Even though all blood donated for transfusion is tested for the presence of human immunodeficiency virus (HIV) antibodies, there exists a period of time after infection by the virus before these antibodies can be detected. Blood donated during this window period is capable of transmitting the virus. Therefore, the blood of persons who are at risk for acquired immune deficiency syndrome (AIDS) should not enter the blood supply. Over a period of 4 months, 6573 potential blood donors who entered fixed and mobile blood collection sites in two cities were exposed to alternative interventions the aim of which was to exclude persons at risk for AIDS. We compared the interventions to one another and to existing materials in terms of the numbers of at‐risk persons who did or did not donate for transfusion, the amount of attention paid to the materials, the scores on a comprehension test, and the self‐ reports by the subjects of attitudes towards the various interventions. At‐risk donors who were asked direct AIDS risk behavior questions in addition to the current health history questions were more likely to be screened out than those who underwent alternative health history interviews (p less than 0.01). Potential donors paid more attention to the experimental brochures than to the experimental video or current materials (p less than 0.05). Comprehension scores were better for the new brochure and the video than for the current brochure (p less than 0.05). Donors were not offended by the experimental interventions. Providing donors with new materials that increase attention and comprehension was not more effective than current materials in screening out potential donors at risk for HIV. Asking donors specific questions about participating in risk behaviors was significantly more effective in screening out at‐risk donors than oral presentation of AIDS information.
White cells protect donor blood against bacterial contamination
Authors: Hogman C.F. , Gong J. , Eriksson L. , Hambraeus A. , Johansson C.S.
Published: March 2003

The possible beneficial role of white cells (WBCs) in donor blood has been investigated with respect to their capacity to remove bacteria. Preparations of buffy coat and whole blood, containing as well as reduced of WBCs, were inoculated with Staphylococcus epidermidis, S. aureus, Escherichia coli, Pseudomonas aeruginosa, and Propionibacterium species. Upon storage at room temperature, the presence of WBCs resulted in a reduction of the bacterial content. Units inoculated with S. epidermidis and E. coli were completely cleared of bacteria within 5 to 24 hours. On the other hand, S. aureus, after an initial reduction in number, started to multiply. In WBC‐reduced units, the initial bacterial content remained unchanged for 5 hours, but the bacteria then exhibited vigorous growth within 48 hours in buffy coat and slower growth in whole blood. Propionibacterium sp. did not grow with or without WBCs. P. aeruginosa did not grow in buffy coat but showed a growth pattern similar to that of S. aureus in whole blood. The presence of WBCs in the donor blood during the first hours after collection thus seems to rid the blood of at least some species of bacteria. These results indicate that it would be favorable not to perform WBC reduction during blood collection and that several hours of contact can be needed to obtain sterility.
Risk factors in hepatitis C virus‐infected blood donors
Authors: van der Poel C. , Cuypers H. , Reesink H. , Choo Q.L. , Kuo G. , Han J. , Quan S. , Polito A. , Verstraten J. , van de Wouw J. , Schaasberg W. , Houghton M. , Lelie P.N.
Published: March 2003

Risk factors of parenteral and nonparenteral exposure to hepatitis C virus (HCV) infection were studied in 125 blood donors in The Netherlands who were positive for anti‐HCV on enzyme‐linked immunosorbent assay (ELISA). Risk factors were related to confirmatory test results of four‐antigen recombinant immunoblot assay (4‐RIBA) and polymerase chain reaction (PCR) of the HCV 5′ untranslated region. Twelve (10%) of the 125 anti‐HCV C100 ELISA‐positive blood donors were 4‐RIBA positive. Eleven (92%) of 12 4‐RIBA‐positive blood donors were PCR positive, and all 113 remaining 4‐RIBA‐negative or ‐indeterminate donors were PCR negative. Eleven (92%) of 12 4‐RIBA‐positive blood donors had a risk factor of parenteral exposure, as compared to 17 (15%) of 113 4‐RIBA‐negative or ‐indeterminate donors. The prevalence of confirmed HCV infection among Amsterdam blood donors is calculated at 0.04 percent; parenteral exposure appears to be the major risk factor for HCV infection.
A randomized trial of perioperative hemodilution versus transfusion of preoperatively deposited autologous blood in elective surgery
Authors: Ness P.M. , Bourke D.L. , Walsh P.C.
Published: March 2003

Hemodilution, one of several methods proposed to decrease homologous blood transfusion in elective surgery, has not been studied in a prospective controlled trial to determine if it is successful. A prospective, randomized controlled study was conducted to determine if hemodilution can serve as an alternative to preoperative autologous blood donation. Fifty patients were randomized to preoperatively deposit 3 units of autologous blood or to undergo hemodilution immediately before elective radical retropubic prostatectomy. All patients were treated under a standard protocol, including surgery performed by a single surgeon. The preoperative deposit groups received a mean of 2.44 +/− 1.0 units of blood; 2 of 25 patients required homologous blood transfusion for blood loss of 2600 mL and 1700 mL. The hemodilution group received a mean of 2.88 +/− 0.4 units of autologous blood: no hemodilution patient received homologous blood. At discharge, the mean hematocrit for the preoperative deposit group was 35.5 +/− 4.9 (0.35 +/− 0.05), and that for the hemodilution group was 31.8 +/− 4.7 (0.32 +/− 0.05) (p less than 0.001). There were no differences in perioperative morbidity in the treatment groups. The best predictor of discharge hematocrit was the initial hematocrit of the patient. It can be concluded that hemodilution can safely replace or at least augment preoperative autologous donations as a means of decreasing homologous blood transfusion in study patients. These results can be applied to any elective surgery procedure in which a 1000‐mL blood loss is anticipated. Other advantages of hemodilution, including convenience, lower cost, and better preservation of all components of autologous blood, suggest that this practice deserves wider application.
Impact of specimen handling and storage on detection of hepatitis C virus RNA
Authors: Busch M.P. , Wilber J.C. , Johnson P. , Tobler L. , Evans C.S.
Published: March 2003

Direct detection of hepatitis C virus (HCV) RNA in serum or plasma is useful for validating the performance of anti‐HCV assays and for the discrimination of persons with persistent HCV infections from those with resolved infections. Quantitation of HCV RNA may also be useful for disease prognosis and therapeutic monitoring. Previous studies have reported detection of HCV RNA in 50 to 70 percent of blood donors who were positive on anti‐HCV supplemental tests. There is concern that specimen processing and storage conditions might influence the stability, and hence the detectability, of HCV RNA. To address this concern, the rate of detection of HCV RNA by the polymerase chain reaction (PCR) using donor pilot tube sera (PTS) previously subjected to routine donor screening and supplemental testing was compared with HCV PCR results obtained with fresh‐frozen plasma (FFP) derived from the same donations. All 16 anti‐HCV supplemental test‐positive donations evaluated were HCV RNA positive with FFP, whereas only 10 (62.5%) were positive with PTS (p = 0.024). None of 11 FFP or PTS samples from HCV enzyme immunoassay‐reactive donations not confirmed by supplemental anti‐HCV assays tested positive for HCV RNA. Direct comparison of sample type (serum vs. plasma) and various storage conditions using specimens from two seropositive donors showed that room‐temperature storage results in marked reduction in HCV RNA signal, while replicate freezing and thawing caused a moderate reduction. These data indicate that well‐controlled sample processing and storage conditions are critical to the sensitive and potentially quantitative analysis of HCV RNA.
Effects of oral donor questioning about high‐risk behaviors for human immunodeficiency virus infection
Authors: Gimble J.G. , Friedman L.I.
Published: March 2003

The purposes of this study were 1) to compare blood donor deferrals resulting from additional, oral questions about human immunodeficiency virus risk behaviors with deferrals resulting from currently used, written screening questions; 2) to examine differences in donor deferral resulting from use of an indirect (IQ) versus direct (DQ) additional oral question format; and 3) to evaluate written survey responses of donors and staff members to the additional questions. The IQ group (n=3050) were asked if they understood the seven ineligible‐donor risk behaviors, and the DQ group (n=4753) were asked if they had engaged in any of these behaviors. Owing to positive answers or refusal to answer the additional questions, there was an increase in donor deferrals, over the level seen with customary screening. Only 1 percent of donors indicated they would not return if the questions were asked in the future. Embarrassment was indicated by 3 percent of the IQ group and 7 percent of the DQ group; 14 to 15 percent preferred to write their answers rather than give them orally. The staff members generally felt training was adequate (IQ=92%, DQ = 83%) and were comfortable asking the questions (IQ=82%, DQ=78%). Mean screening times were 5.7 minutes before the addition of the oral questions, 7.5 minutes with IQ, and 7.6 minutes with DQ. This study confirms the value of IQ and DQ formats in identifying potentially infectious donors and suggests that the DQ format may be slightly more effective.
In vitro storage and in vivo survival studies of red cells from persons with the In(Lu) gene
Authors: Ballas S.K. , Marcolina M.J. , Crawford M.N.
Published: March 2003

Red cells (RBCs) of individuals with the In(Lu) gene are characterized by suppression of the Lutheran, P1, i, and other blood group antigens, acanthocytosis, and abnormal electrolyte metabolism. To determine the clinical significance of these abnormalities, the survival of autologous RBCs was determined by 51Cr in two siblings with the dominant Lu(a‐b‐) [In(Lu)] phenotype. Both subjects studied had normal hemoglobin, hematocrit, reticulocyte count, haptoglobin, and ferritin values. RBC indices were mildly hypochromic. Examination of the peripheral smear showed mild acanthocytosis in one individual. Analysis of RBC distribution on discontinuous density gradients showed a shift to lighter fractions than normal control RBCs. Storage of these Lu (a‐b‐) RBCs at 4 degrees C showed significant hemolysis within a few days; this was confirmed by increased autohemolysis, which was reduced by glucose and ATP. RBC cation content (sodium and potassium) was higher than that in control cells, which indicated increased cell hydration, which explains the lighter density and mild hypochromia of the Lu(a‐b‐) RBCs. 51Cr survival of autologous Lu(a‐b‐) RBCs was normal in both subjects studied. The data indicate that the morphologic and cation abnormalities of RBCs of persons with the In(Lu) gene are clinically insignificant, as these cells have normal in vivo survival. Such RBCs, however, are susceptible to increased hemolysis in vitro under standard blood banking storage conditions. Individuals of the Lu(a‐b‐) phenotype, associated with In(Lu), may not be suitable candidates for routine blood donation.
Thrombotic thrombocytopenic purpura: outcome in 24 patients with renal impairment treated with plasma exchange
Authors: ROCK G. , SHUMAK K. , KELTON J. , BLANCHEITE V.S. , BUSKARD N. , NAIR R. , SPASOFF R. ,   CANADIAN APHERESIS STUDY GROUP
Published: March 2003

The Canadian Apheresis Study Group recently completed a randomized clinical trial involving 102 patients with thrombotic thrombocytopenic purpura (TTP), in which treatment with plasma infusion and treatment with plasma exchange were compared. Thirty‐three other patients were ineligible or refused to be randomly assigned in the trial. Of the 33 patients, 24 were assessed as ineligible because they would be unable to tolerate the fluid input that would occur if they were randomly assigned to receive plasma infusion. All 24 patients had oliguria and elevated creatinine and/or blood urea nitrogen level. These 24 patients were treated with acetylsalicyclic acid, dipyridamole, and plasma exchange according to the standardized protocol defined in the trial. Blood for tests of factors possibly involved in the pathogenesis of TTP was drawn before exchange and at intervals during and after exchange. The mean platelet count before exchange was 35.5 × 10(9) per L. In 12 of the 24 patients, the platelet count reached 150 × 10(9) per L or greater by 7 days after the initiation of plasma exchange. Three patients responded partially, in that their platelet count increased to at least twice that at presentation, but remained below 150 × 10(9) per L. One patient died during the first week. Of the eight other patients who experienced treatment failure at the 7‐day assessment point, six subsequently responded, four while continuing to receive plasma exchange and two after plasma exchange had been discontinued. Of the 15 patients who either responded fully or responded partially by the end of the first cycle, all survived.(ABSTRACT TRUNCATED AT 250 WORDS)
Increased detection of hepatitis C virus (HCV)‐infected blood donors by a multiple‐antigen HCV enzyme immunoassay
Authors: KLEINMAN S. , ALTER H. , BUSCH M. , HOLLAND P. , TEGTMEIER G. , NELLES M. , LEE S. , PAGE E. , WILBER J. , POLITO A.
Published: March 2003

A new, multiple‐antigen enzyme immunoassay (EIA‐2) for hepatitis C virus (HCV) antibodies was evaluated in parallel with the previously available c100‐3 HCV EIA (EIA‐1) in 14,068 volunteer blood donors as well as in 25 cases of transfusion‐associated hepatitis C for which recipient and donor samples were available. When compared to EIA‐1, the EIA‐2 was more sensitive in detecting HCV‐infected blood donors. The EIA‐2 detected an additional 1 in 1000 EIA‐1‐negative, surrogate marker‐ negative donors who were infected with HCV as demonstrated by polymerase chain reaction (PCR). The specificity of the EIA‐2 was comparable to that of the EIA‐1, but the two tests appear to detect different populations of false‐positive donors. Recombinant immunoblot assay‐indeterminate donors were detected five times more frequently by the EIA‐2; PCR demonstrated that 21 percent of these donors were infected with HCV. The greater sensitivity of EIA‐2 was also found in 25 transfusion recipients with non‐A, non‐B hepatitis; however, in 16 percent of these cases of posttransfusion HCV infection, the EIA‐2 failed to detect an HCV‐seropositive donor. These data indicate that EIA‐2 testing will significantly reduce, but probably not eliminate, the risk of transfusion‐associated HCV infection; we estimate this residual per‐unit risk to be 1 in 2000 to 1 in 6000 units transfused. On a national level, it is projected that the replacement of the anti‐ HCV EIA‐1 with the EIA‐2 will initially prevent up to 40 additional cases of transfusion‐associated hepatitis C per day.
Analysis of a linear peristaltic infusion device for the transfusion of red cells to pediatric patients
Authors: Criss V.R. , DePalma L. , Luban N.L.
Published: March 2003

A linear peristaltic infusion device was evaluated for red cell (RBC) transfusion in the pediatric and neonatal setting. CPDA‐1 RBC units (n = 24) divided into six groups of 4 units each underwent simulated transfusion. Blood was infused by using manufacturer‐provided administration sets with either a 21‐gauge needle or a 24‐gauge catheter. Filters were used in two groups to evaluate the effect of negative pressure on filter function. Two groups of RBCs less than 1 week old were washed, irradiated, and infused at 5 mL per hour, by using a standard administration set, or at 10 mL per hour, by using a syringe set. Four‐week‐old RBCs (washed and irradiated, irradiated and filtered, filtered only, or unmanipulated) were infused at 100 mL per hour. Paired samples from 0 and 2 hours before and after infusion were analyzed for hemoglobin, hematocrit, RBC count, plasma hemoglobin, lactate dehydrogenase, potassium, alanine aminotransferase, and aspartate aminotransferase. Hausser and Nageotte hemocytometers were used to perform white cell (WBC) counts when a filter was used. By analysis of variance and percentage of change, data from 0 and 2 hours before and after infusion were compared. No clinically or statistically significant differences were seen for hemoglobin, hematocrit, or RBC count. The difference in preinfusion and postinfusion plasma hemoglobin levels in washed RBCs at 2 hours was statistically but not clinically significant (14.5 ± 6.8 vs. 19.3 ± 7.1 mg/dL). No clinically significant differences were noted for the remaining analytes. In filtered RBCs at 2 hours, the WBC removal was 3.08 ± 0.28 log10 which resulted in fewer than 2 times 106 WBCs per unit. The linear peristaltic infusion device is suitable for use in a pediatric setting.
Cytokine production in IgG‐mediated red cell incompatibility
Authors: Davenport R.D. , Burdick M. , Moore S.A. , Kunkel S.L.
Published: March 2003

The transfusion of incompatible red cells may result in fever and systemic symptoms. The mechanisms by which these symptoms are produced in the setting of antibodies that do not usually fix complement, as in the Rh system, are obscure. It has been hypothesized, on the basis of their known biologic activities, that a specific set of cytokines may be involved in such transfusion reactions. Therefore, the production of the inflammatory cytokines interleukin‐1 beta (IL‐1 beta), tumor necrosis factor (TNF), interleukin‐6 (IL‐6), and interleukin‐8 (IL‐8) by human monocytes in response to red cells sensitized with anti‐D was investigated, as a model of IgG‐dependent hemolytic transfusion reactions. IL‐1 beta, IL‐6, and IL‐8 were detectable in the culture supernatants at 4 to 6 hours and increased up to 24 hours, whereas TNF peaked at 6 hours. Immunocytochemical stains of cell preparations demonstrated IL‐1 beta, IL‐8, and TNF in monocytes engaged in erythrophagocytosis. IL‐8 production and phagocytosis could be inhibited by monomeric IgG, but Fab fragments of a monoclonal antibody specific for the low‐affinity IgG receptor Fc gamma RII could not be, which suggests the involvement of the high‐affinity receptor Fc gamma RI. Neutralizing antisera to IL‐1 beta and TNF did not abrogate the production of IL‐8, which suggests that sensitized red cells serve as a primary signal for this cytokine. These findings indicate that the production of inflammatory cytokines by phagocytes may be responsible for the symptomatology of IgG‐mediated hemolytic transfusion reactions.
Polymerase chain reaction‐based detection of MN blood group‐specific sequences in the human genome
Authors: Corfield V. A. , Moolman J. C. , Martell R. , Brink P. A.
Published: March 2003

The MN blood group antigens have traditionally been detected by serotyping; however, development of a DNA‐based method offers flexibility in the determination of this highly polymorphic system. Genotyping the MN blood group antigens was performed by polymerase chain reaction amplification of the specific alleles (PASA) in the human genome. In separate paired reactions, M or N allele‐specific oligonucleotide primers were amplified with a common distal primer. Only in the presence of the homologous template was a 781‐base pair polymerase chain reaction amplification product visible after agarose gel electrophoresis and ethidium bromide staining. This method of genotyping could be performed using either 1 μg of extracted DNA or 0.5 microL of whole blood, and the results showed 100‐percent correlation with those obtained by serotyping. PASA‐based genotyping of MN blood group antigens, which requires a small amount of starting material, has application in linkage and population studies and in forensic medicine.
A retrospective study to determine the risk of red cell alloimmunization and transfusion during pregnancy
Authors: Heddle N.M. , Klama L. , Frassetto R. , O'Hoski P. , Leaman B.
Published: March 2003

A retrospective review of post‐delivery antibody records was performed at a teaching hospital and a community hospital to determine the frequency of new red cell alloantibody production and transfusion during pregnancy. If alloantibody was undetected at delivery, it was assumed that alloimmunization had not occurred. When antibody was detected, a chart review was performed to determine if the antibody was present at the beginning of the pregnancy or was newly produced during the pregnancy. A total of 17,568 pregnancies were reviewed. Antibody was detected at delivery in 948 (5.4%) cases, of which 89.5 percent (848/948) involved passive anti‐D or clinically insignificant antibodies. The remaining 100 pregnancies involved clinically significant IgG antibodies. In 58 pregnancies, the antibody was detected in the first trimester, and in 42, new antibody production occurred during the pregnancy. Thus, the prevalence of new antibody production during pregnancy was 0.24 percent (95% confidence interval [CI], 0.17‐0.32). Transfusion records indicated that the prevalence of transfusions during pregnancy was 0.09 percent (95% CI, 0.04‐0.14). None of the women with new alloantibody formation during their pregnancies required transfusion; hence, new alloantibody production and the need for transfusion appear to be independent events. The probability of these events occurring together was 2.1 × 10(‐6), or 1 in 500,000 deliveries.
CD34+ progenitors and colony‐forming units‐granulocyte macrophage are recruited during large‐volume leukapheresis and concentrated by counterflow centrifugal elutriation
Authors: Hillyer C.D. , III D.A. Lackey , Hart K.K. , Stempora L.L. , Bray R.A. , Bender J.G. , Donnenberg A.D.
Published: March 2003

The recruitment of mononuclear cells (MNCs), colony‐forming units‐ granulocyte macrophage (CFU‐GM), lymphocyte subpopulations, and CD34+ progenitor cells was studied during large‐volume (15–25 L blood processed) peripheral blood stem cell (PBSC) harvests. Normal donors (n = 13) underwent a 4‐hour leukapheresis designed to maximize PBSC yield (blood flow rate, 85 mL/min). Mean (± SD) volume processed was 17.7 ± 0.4 L, and yield was 2.4 ± 0.7 × 10(10) white cells containing 99 percent MNCs and 1.3 mL red cells per L of blood processed. Postapheresis hematocrit, platelets, and MNCs were reduced from preapheresis values by 7, 35, and 23 percent, respectively (p < 0.05). In nine donors, the component was collected as four 1‐hour samples, and culturing of CFU‐GM and flow cytometric analysis of lymphocyte subpopulations and CD34±;/HLA‐DR±; cells were done in individual samples. Total CFU‐GM were 2.4 ± 1.4 × 10(6) (3.0 ± 1.8 × 10(4) CFU‐GM/kg) and lymphocytes were 20.8 × 10(9), with 75 percent CD3±; T cells, 10 percent CD19/CD20±; B cells, and 17 percent natural killer cells. A more than twofold increase in CFU‐GM and CD34±; cells was noted over the course of the 4‐hour procedure (p < 0.05). In four donors, the leukapheresis component underwent counterflow centrifugal elutriation (CCE), which separated it into four fractions in an attempt to concentrate CD34±; and CFU‐GM progenitors and to deplete T‐lymphocytes on a large scale. There was a 1.8‐, 4.6‐, 3.9‐, and 0.32‐fold increase in CFU‐GM in the four fractions relative to the unseparated component. CD34+ cells represented 0.09,0.23, 0.66, and 0.41 percent of the fractions, respectively, while there was a relative de‐ pletion of CD3+ T‐lymphocytes by 61.4, 78.7, 85.1, and 98.1 percent. Thus, MNCs, CFU‐GM, and CD34+ cells are recruited during lar e‐volume leukapheresis, which ma generate adequate mean CFU‐GM er kg to afow PBSC bone marrow rescue in &wer procedures. CD34+ and CFU‐eM rogenitors are concentrated by CCE, and T cells are modestly depleted by CCE.
A microplate system for ABO and Rh(D) blood grouping
Authors: Chung A. , Birch P. , Ilagan K.
Published: March 2003

A microplate system for performing ABO and Rh(D) blood group determinations with a Kemble Kemtek 1000SP liquid handling processor, an Anthos 2001 microplate reader, an IBM Personal System 2 microcomputer, and Sanguin Forma software is described. The performance of this Kemble/Anthos/IBM/Sanguin microplate system for ABO and D grouping was evaluated by testing 10,042 routine blood donor samples in parallel with the forward‐grouping channels of a Technicon AutoAnalyzer blood grouping system. Manual techniques were used to perform ABO reverse groupings and to resolve all ABO forward‐ and reverse‐grouping discrepancies. D‐negative test results were investigated and confirmed manually by the indirect antiglobulin test. Of the 10,042 samples tested, 97.3 percent were grouped correctly. There were 266 samples whose results were flagged as no type determined, of which 124 were ABO tests and 142 were Rh tests. In addition, 30 weak D samples missed by both automated systems were detected by manual indirect antiglobulin test. The system is flexible and easy to maintain and operate.
Plateletpheresis: comparison of platelet yields, processing time, and white cell content with two apheresis systems
Authors: Burgstaler E.A. , Pineda A.A. , Brecher M.A.
Published: March 2003

Two apheresis systems (COBE Spectra and Fenwal CS‐3000 Plus with TNX‐6 chamber [CS‐3000 Plus]) were compared by using their yield predictors and maximum processing times in regard to platelet yields and processing times (n = 200 each). Platelet yields (n = 50 each) and white cell (WBC) content (n = 20 each) from procedures using the current software revision (3.6) on the Spectra and various interface offset (IO) settings on the CS‐3000 Plus were also compared. Significantly higher median platelet yields (4.88 [range, 1.84‐9.97] vs. 4.57 [range, 2.82‐9.20] × 10(11) platelets) and frequency of components with > or = 6.0 × 10(11) platelets (31.5 vs. 21.5%) were found with the Spectra. In the study with Spectra and the CS‐3000 Plus IO settings, IO‐10 and IO‐13, overall, produced the most platelets, although the differences were not significant. All Spectra collections tested had < 5 × 10(8) WBCs, and, depending on the software revision, had either 85 percent (revision 3.6) or 100 percent (revision 2.5) of components with < 5 × 106 WBCs. To ensure that 100 percent of components contained < 5 × 108 WBCs when the CS‐3000 Plus was used, IO settings of 10 or less were required, and to ensure components with < 5 × 10(6) WBCs, only IO‐6 could be used. Spectra and CS‐3000 Plus were capable of collecting large doses of platelets (up to the equivalent of 18 units) in processing times of 67 to 100 minutes with < 5 × 106 WBCs.