bluuebunny/crossref_metadata_2025 huggingface dataset:
The changing activities of a regional marrow donor program
Authors: Stroncek D. , Strand R. D. , Hofkes C. L. , McCullough J.
Published: March 2003

BACKGROUND: A regional marrow donor program was established in 1982. Following the establishment of the National Marrow Donor Program (NMDP) in 1987, the activities of this regional program changed. STUDY DESIGN AND METHODS: To better understand the changes that occurred in the regional marrow donor program, its donor recruitment and marrow collection activities through 1991 were studied retrospectively. Data analyzed included the total number of potential donors, the number and types of potential donors recruited each year, the number of searches performed, the number of samples collected for HLA‐DR typing and mixed lymphocyte culture testing, and the number of transplants in both programs from 1987 through 1991. Statistical analysis was performed by using chi‐square. RESULTS: Initially, only persons who donated platelets by apheresis were enrolled into the program. In 1986, the regional program's first drive to recruit people who were not apheresis donors occurred. The number of such drives increased each year, and in 1991, 12 drives occurred, which resulted in the recruitment of 1313 potential marrow donors. From 1987 to 1991, the number of potential donors in the regional program grew from 3252 to 9146, but the proportion of apheresis donors in the program decreased. In 1987, 91.9 percent of marrow donors at the regional center had been apheresis donors, but in 1991, 41.7 percent had been apheresis donors. The number of marrows donated at the regional center increased from 11 in 1987 to 29 in 1989, but then fell to 24 per year in 1990 and 1991. The decrease in the number of donations at the regional program was due to the rapid growth in the NMDP file of potential marrow donors and the selection of donors whose HLA antigens were more compatible with those of the transplant recipients. In 1989, the regional program contained 4.6 percent of all HLA‐A,B‐typed and 11.2 percent of all HLA‐A,B,DR‐typed potential donors in the NMDP and collected 15.3 percent of all marrows. However, in 1991, the regional program contained 2.0 percent of HLA‐A,B‐ typed donors and 4.1 percent of HLA‐A,B,DR‐typed donors and collected 5.3 percent of marrows. In 1987, 18 percent of the people who donated marrow at the regional center were phenotypically HLA‐A,B,DR identical with the recipient, but in 1991, 92 percent of donor‐recipient pairs were phenotypically HLA‐A,B,DR identical. CONCLUSION: Recruitment activities became an increasingly larger part of the Regional Marrow Donor Program's activities. Increasing the size of the file of potential donors was necessary to maintain a constant number of donations. Persons who were not regular blood donors were an important part of the marrow donor program.
Electronic verification of donor‐recipient compatibility: the computer crossmatch
Authors: Butch S.H. , Judd W.J. , Steiner E.A. , Stoe M. , Oberman H.A.
Published: March 2003

Background: This article describes standard operating procedures (SOPs) for a computer crossmatch to replace the immediate‐spin crossmatch for ABO incompatibility between patient blood samples submitted for pretransfusion testing and the blood component selected for transfusion. These SOPs were developed following recent changes to the Standards for Blood Banks and Transfusion Services of the American Association of Blood Banks (AABB).

Study Design and Methods: SOPs were developed, utilizing currently available software, for pretransfusion testing. The SOP for donor unit processing entails bar code entry of the unit number, component name, and ABO/Rh type; computer entry and interpretation of serologic reactions; warning of discrepancies between bar code‐entered blood type and result interpretation; and quarantine of the donor unit in such instances. The SOP for patient sample testing requires bar code entry of specimen accession number, which accesses patient demographics; computer entry and interpretation of ABO/Rh tests; repeat blood typing at the time of crossmatch if only one patient blood type is on record; and warning if there are nonconcordant current and historical blood types. The computer crossmatch SOP requires bar code entry of specimen accession and donor unit numbers; release of group O red cells pending resolution of discrepancies; and immediate‐spin crossmatch during computer downtime. Tables validated on‐ site prompt warning messages and prevent both computer crossmatch and release if blood components of the wrong ABO type are selected.

Results: These SOPs meet the requirements of the 15th edition of the AABB Standards. Projected annual time savings at this institution are > 100,000 workload recording units. Further benefits include reduced patient sample volume requirements, less handling of biohazardous material, and elimination of unwanted positive or negative reactions associated with the immediate‐spin crossmatch. Release of incompatible blood components when the wrong patient blood type is on record is addressed by requiring the use of group O red cells in the absence of two concordant blood types, one of which must be from a current sample.

Conclusion: A combination of existing computer programs and carefully developed SOPs can provide a safe and efficient means of detecting donor‐recipient incompatibility without performance of serologic crossmatch.
Potential deferral criteria predictive of human immunodeficiency virus positivity among blood donors in Thailand
Authors: Kitayaporn D. , Bejrachandra S. , Chongkolwatana V. , Chandanayingyong D. , Weniger B.G.
Published: March 2003

Background: To develop deferral criteria to prevent human immunodeficiency virus (HIV) transmission by recently infected blood donors in the seronegative “window” phase, routine data on donors at a university hospital were examined for factors predicting seropositivity.

Study Design and Methods: Records of all 281 HIV‐ positive blood donors from August 1987 through September 1991 were retrospectively compared with those of 1076 randomly selected control donors matched only by year of donation. Four controls were selected for each HIV‐positive donor.

Results: The prevalence of HIV in 102,684 donor units during the period rose from 0.02 percent in 1987 to 0.52 percent in 1991. Multivariable analysis revealed that male sex (odds ratio [OR] = 26.4), VDRL test positivity (OR = 3.0), age 21 to 30 years (OR = 2.2; referent: 16–20‐year‐old group), and replacement donorship (OR = 1.4; referent: voluntary donors) were independent factors significantly associated with HIV positivity among these donors (p < 0.05). Since replacement donorship cannot be avoided, only male sex, age 21 to 30 years, and VDRL test positivity were considered as potential criteria. When these findings were extrapolated to all donors in 1990 and 1991, those with all three or only two (excluding VDRL test, because the results are known only after donation) of these high‐ risk factors had HIV positivity probabilities of 2.2 and 1.0 percent, respectively. These probabilities were, respectively, 4.9 times (95% CI: 2.9 8.3) and 4.1 times (3.1, 5.4) the risk among other donors. However, applying such criteria would have eliminated 1.5 and 31.2 percent, respectively, of all HIV‐negative donors in 1990 and 1991. The latter deferral proportion is too high to be acceptable.

Conclusion: In Thailand, improved donor deferral criteria addressing sexual risk factors could lead to decreased probability of window‐period donation, with an acceptable rate of deferral. Additional p24 antigen testing may be indicated for donors at increased risk for HIV infection, specifically, men aged 21 to 30.
Incest diagnosis by comparison of alleles of mother and offspring at highly heterozygous loci
Authors: Wenk R.E. , Chiafari F.A. , Houtz T.
Published: March 2003

Background: Methods that detect a child's homozygosity by examination of allelic products are insensitive for diagnosing incest because, at a given locus, a homozygous state is expected with a frequency of only 0.25 when parents are first‐degree relatives. Furthermore, these methods are not specific if the population contains many homozygous individuals or silent alleles that cause apparent homozygosity.

Study Design and Methods: Use of highly heterozygous loci improves specificity, but not sensitivity. Sensitivity may be increased by observing for two kinds of mother‐offspring similarities: an offspring of incest tends to be homozygous or heterozygous‐identical with respect to its mother's phenotype. At each locus, two conditional probabilities may be calculated for a genetic observation, using allele frequencies expected under a state of incestuous mating versus mating within a specified population. The conditional probabilities at each locus are compared in a likelihood ratio to express a relative probability of incest.

Results: In a case of known sibling incest, three likelihood ratios were derived from variable number of tandem repeat phenotypes at five loci. When only offspring homozygosity was observed, the likelihood ratio was 75.3:1. When both homozygous‐ and heterozygous‐ identical phenotype similarities of offspring and mother were noted, the likelihood ratio was 130.4:1. When maternal obligatory alleles of the offspring were considered, the likelihood ratio was 262.4:1.

Conclusion: Comparison of maternal and offspring phenotypes at highly heterozygous loci increases both sensitivity and specificity of genetic tests in cases of suspected incest.
Evaluation of clinical and laboratory aspects of antibody tests for detection of hepatitis C virus infection in blood donors and recipients from a low‐risk population
Authors: MacDonald K.L. , Mills W.A. , Wood R.C. , Hanson M. , Kline W. , Bowman R.J. , Polesky H.F. , Williams A.E. , Osterholm M.T.
Published: March 2003

Background: When the first‐generation enzyme immunoassay (EIA) for detection of antibody to hepatitis C virus (anti‐HCV) was approved in May 1990, blood banking agencies recommended testing of all components in inventory. In many cases, one or more components from these units had already been transfused.

Study Design and Methods: Donors that reacted in first‐generation EIAs and recipients of their components were identified, and anti‐HCV test methods (including first‐generation EIA, second‐generation EIA, and recombinant immunoblot assay [RIBA]) were evaluated.

Results: Of 66 donors identified as anti‐HCV‐positive by first‐generation EIA, 17 were positive in second‐generation EIA. Of these 17, 9 reacted in RIBA; 6 of these showed evidence of HCV infection in polymerase chain reaction (4) and/or probable transmission of HCV to a transfusion recipient (3). Of the 48 specimens that were positive in first‐generation EIA and negative in second‐generation EIA, only 1 was positive in RIBA; serum was not available for polymerase chain reaction testing, and there were no living transfusion recipients in whom to assess evidence of transmission of HCV.

Conclusion: This study documents the low predictive value of EIAs for anti‐HCV in a low‐ prevalence blood donor population and emphasizes the need for additional testing to confirm the specificity of samples that react in the screening tests.
Pseudoaneurysm of the brachial artery: a rare complication of blood donation
Authors: Popovsky M.A. , McCarthy S. , Hawkins R.E.
Published: March 2003

Background: Serious complications of blood donation are rare. Reported here is a significant vascular event that resulted from whole‐blood phlebotomy and required surgical repair.

Case Report: A 49‐year‐old woman donated whole blood without incident in November 1992, but reported ecchymosis and tenderness in the antecubital area of the right arm immediately following the procedure (her medical history was unremarkable). Over the next several days, the ecchymosis extended to the axilla and she complained of numbness in all her fingers, particularly the thumb and adjacent two fingers of the involved side. Coincident to these findings, an egg‐sized lump developed under the skin at the phlebotomy site. Her symptoms and the mass persisted for several weeks. Examination and ultrasound performed in January 1993 demonstrated a pulsatile mass over the brachial artery. The donor underwent an exploratory surgical procedure that revealed a 3.3 × 2.5 × 2.0‐cm pseudoaneurysm of the brachial artery. Unlike a true aneurysm, this lesion was composed only of a blood clot that was compressing the outer wall of the artery. A pinpoint‐sized hole was seen in the artery when the lesion was excised. The phlebotomy is believed to have caused blood to leak out of the artery, creating the lesion and exerting pressure on the median nerve.

Conclusion: Pseudoaneurysm of the femoral artery is a well‐documented complication of cardiac catheterization and other invasive arterial procedures, but this is the first known reported case following blood donation. Blood collectors should be made aware of this potentially serious complication of blood donation.
Identification of a new white cell antigen
Authors: STRONCEK D.F. , RAMSEY G. , HERR G.P. , EIBER G. , CLAY M.E. , KETYER E.C.
Published: March 2003

BACKGROUND: Antibodies to white cell antigens can cause alloimmune neonatal neutropenia, autoimmune neutropenia, and transfusion reactions.

CASE REPORT: A full‐term male infant developed a skin infection and was found to be neutropenic on his fourth day of life. He had a transient increase in his neutrophil count after treatment with intravenous immunoglobulin, but his neutrophil count was not consistently normal until he was 6 weeks old. Serum from the baby's mother reacted in a granulocyte immunofluorescence assay but not in a granulocyte agglutination assay. The mother's serum was tested in the granulocyte immunofluorescence assay against neutrophils from 103 healthy, unrelated people, and it reacted with cells from 66 percent of those people. The expression of SL correlated weakly with the expression of NA1 (r = 0.23; p = 0.02) and 5a (r = 0.20; p = 0.05) antigens. SL antigen expression on neutrophils was not associated with the expression of NA2, NB1, NB2, NC1, 5b, 9a, or Mart. The expression of SL on neutrophils from members of an extended family was analyzed, and the antigen was found to be inherited in an autosomal‐dominant manner. Anti‐SL also reacted with T‐lymphocytes in a flow cytometry assay but did not react with red cells or platelets. No lymphocytotoxic antibodies were detected in the mother's sera. The anti‐SL was tested against neutrophils in an immunoprecipitation and immunoblotting assay, but no molecules were identified. The neutrophil‐specific antigens NA are located on Fc gamma receptor III (CD16). To determine if the SL antigen was also located on Fc gamma receptor III, anti‐SL was also tested in a monoclonal antibody immobilization of granulocyte antigens assay. Anti‐SL did not react with molecules recognized by CD16 monoclonal antibodies.

CONCLUSION: A new white cell antigen SL, with a frequency of 66 percent, was identified on neutrophils and T‐ lymphocytes as a result of the evaluation of a case of neonatal alloimmune neutropenia. The molecule bearing the SL antigen was not identified in immunoblotting, immunoprecipitation, or monoclonal antibody immobilization of granulocyte antigens assays.
An evaluation of intravenous immunoglobulin in the treatment of human immunodeficiency virus‐associated thrombocytopenia
Authors: JAHNKE L. , APPLEBAUM S. , SHERMAN L.A. , GREENBERGER P.A. , GREEN D.
Published: March 2003

BACKGROUND: Anecdotal evidence suggests that high‐dose intravenous immunoglobulin (IVIG) is useful in the management of human immunodeficiency virus (HIV)‐associated thrombocytopenia.

STUDY DESIGN AND METHODS: To rigorously evaluate this therapy, a crossover study was designed to compare IVIG, given at 1 g per kg per day for 2 consecutive days each week for 4 weeks, with intravenous saline placebo administered according to the same schedule. Subjects were randomly assigned to receive either IVIG or saline during the first 4 weeks; if IVIG was given, there was a 4‐week period of no therapy before beginning placebo administration. Criteria for eligibility were platelet count of less than 50,000 per microL (50 × 10(9)/L), elevated platelet‐associated IgG levels, increased megakaryocytes in the bone marrow, and positive HIV antibody test. Twelve patients (11 men, 1 woman) were studied. Seven patients completed the full protocol. Four dropped out: after 2, 5 (2 patients), and 8 weeks that included at least 2 weeks of IVIG.

RESULTS: All patients sustained an increase in platelet count in response to IVIG, with increments ranging from 15,000 to 358,000 per microL (15 to 350 × 10(9)/L) (mean, 180,000/microL [180 × 10(9)/L]; median, 174,000/microL [174 × 10(9)/L]). No patient had an increase after placebo infusions. There were no adverse effects of treatment, and weekly chemical analyses showed no new abnormalities except for mild elevations in the serum protein. The duration of responses ranged from 2 to 10 weeks. No patient demonstrated refractoriness to IVIG.

CONCLUSION: IVIG consistently raises platelet counts in patients with HIV‐associated thrombocytopenia.
Molecular basis of the K:6,‐7 [Js(a+b−)] phenotype in the Kell blood group system
Authors: Lee S. , Wu X. , Reid M. , Redman C.
Published: March 2003

BACKGROUND: The Kell blood group system consists of at least 21 antigens. KEL6(Jsa) is a low‐incidence antigen that has an antithetical relationship with the high‐incidence KEL7(Jsb) antigen. The molecular basis of KEL6 that appears in less than 1.0 percent of the general population, but in up to 19.5 percent of African Americans, was unknown.

STUDY DESIGN AND METHODS: Nineteen exons of the Kell gene (KEL) were amplified by polymerase chain reaction (PCR) assays of genomic DNA obtained from individuals with K:6,‐7 [Js(a+b‐)] phenotype. The PCR products were sequenced. A comparison was made of the sequence of the PCR products and the sequence of K:‐6,7, the common phenotype.

RESULTS: KEL from individuals with the K:6,‐7 phenotype had two base substitutions in exon 17. One was a missense mutation (T‐to‐C base substitution) at nucleotide (nt) 1910, which predicts an amino acid change from leucine to proline; the other was a silent substitution (A‐ to‐C) at nt 2019. The T‐to‐C substitution eliminated a restriction site for Mnl I, whereas the A‐to‐G substitution eliminated a Dde I site. Analyses of exon 17 in seven unrelated persons with K:6,‐7 phenotype by Mnl I and Dde I enzymes showed the expected presence of restriction fragment length polymorphisms.

CONCLUSION: The base substitutions T‐to‐ C at nt 1910 and A‐to‐G at nt 2019 are unique to KEL6. The predicted Leu–>Pro change may disrupt the alpha‐helical structure and thus form the epitope for KEL6.
Declining value of alanine aminotransferase in screening of blood donors to prevent posttransfusion hepatitis B and C virus infection. The Retrovirus Epidemiology Donor Study
Authors: Busch M.P. , Korelitz J.J. , Kleinman S.H. , Lee S.R. , AuBuchon J.P. , Schreiber G.B.
Published: March 2003

BACKGROUND: Since the mid‐1980s, blood banks in the United States have screened donors for elevated alanine aminotransferase (ALT) in an effort to prevent posttransfusion hepatitis. The present study was designed to quantitate the residual value of ALT screening following the implementation of hepatitis C virus (HCV) assays.

STUDY DESIGN AND METHODS: Two approaches were used. First, a database of 2.3 million donations made by 586,507 volunteer blood donors between 1991 and 1993 was used to compare the incidence of seroconversion to hepatitis B virus (HBV) and HCV marker positivity in donors with elevated ALT values and with normal ALT values. Second, the duration of ALT elevation prior to HBV and HCV seroconversion was determined from 34 well‐documented cases of posttransfusion HBV and HCV; elevated‐ALT window periods were multiplied by rates of HBV and HCV incidence in donors to project the yield of ALT screening. Predictive value and cost‐ effectiveness analyses were also performed to compare the value of ALT screening before and after HCV screening was implemented.

RESULTS: Both approaches indicate that ALT testing does not detect HBV in the window phase but does currently identify approximately 3 HCV window‐phase donations per 1 million donations; this contrasts with ALT detection of approximately 1800 HCV‐infectious units per 1 million donations prior to anti‐HCV screening. Currently, only 8 in 10,000 donated units with elevated ALT (negative anti‐HCV) are infected with HCV. The cost of continued ALT screening was estimated at $7,931,000 per quality‐ adjusted year of life saved.

CONCLUSION: The yield, predictive value, and cost‐effectiveness of ALT screening of blood donors have declined dramatically with the implementation of progressively improved anti‐HCV assays. ALT screening of volunteer blood donors should be discontinued.
The protein composition of red cell eluates
Authors: Dumaswala Umakant J. , Sukati Hosea M. , Greenwalt Tibor J.
Published: March 2003

BACKGROUND: The protein composition of red cell (RBC) eluates has not been extensively studied. The purpose of this study was to determine the IgG content and protein composition of RBC eluates prepared by the acid and xylene elution methods. STUDY DESIGN AND METHODS: Six samples of group O R1R1 RBCs sensitized with anti‐D in vitro, six nonsensitized samples of the same group O R1R1 RBCs, and six samples from patients with warm autoimmune hemolytic anemia (WAIHA) were studied. The eluate protein composition was determined by sodium dodecyl sulfate‐ polyacrylamide gel electrophoresis, and immunoglobulin concentrations were estimated by an immunoblot technique using horseradish peroxidase‐ conjugated anti‐IgG and 3,3′ diaminobenzidine. RESULTS: The protein concentrations of the xylene eluates were significantly greater than those of the acid eluates (37.3 +/− 10.7 and 3.0 +/− 0.4 [SD], respectively; p < 0.005). In all samples, the proportion of IgG was less than 0.13 percent of the total protein content. The acid eluates of sensitized RBCs contained more IgG than the xylene eluates. The antibody titers of eluates from WAIHA RBCs were significantly lower than eluates of in vitro sensitized RBCs (p < 0.005). The estimated molecular weights of the Coomassie blue‐stainable protein bands from xylene eluates were 97, 78, 63, 45, 31, 23, and 16 kDa, and those of bands from acid eluates were 97, 78, and 55 kDa. No periodic acid‐ Schiff reagent‐stainable bands were detected. CONCLUSION: These data indicate that IgG represented only a small fraction of the proteins in the eluates and that the protein composition varies with the elution procedure.
Construction of bacteriophage expressing mouse monoclonal Fab fragments directed against the human MN glycophorin blood group antigens
Authors: Czerwinski M. , Siegel D. L. , Moore J. S. , Spitalnik P. F. , Spitalnik S. L.
Published: March 2003

BACKGROUND: The MN human blood group antigens are complex glycopeptide antigens at the amino terminus of glycophorin A. Many different mouse monoclonal antibodies to these antigens have been produced and characterized. The construction of combinatorial immunoglobulin libraries displaying antibody Fab fragments on the surface of bacteriophage (Fab‐phage) represents a novel approach for developing monoclonal reagents, for exploring the diversity of the immune response to specific antigens, and for understanding the molecular basis of the interaction of an antibody with its antigen. However, it is necessary to determine whether Fab fragments displayed on bacteriophage surfaces retain immunologic characteristics similar to the intact antibodies. STUDY DESIGN AND METHODS: Fab‐phage were constructed from three anti‐N (AH7, N61, and N92) and two anti‐M (425/2B and M2A1) murine hybridomas. The Fab‐phage and parental hybridomas were compared by enzyme‐linked immunosorbent assay, Western blotting, and flow cytometry. RESULTS: In each case, the Fab‐phage and its parental hybridoma antibody had similar immunologic characteristics. In particular, their dependence on the pH of the buffer and on sialylation of the target antigen was similar. CONCLUSION: These results suggest that Fab‐phage may provide novel reagents with applications in immunohematology and may be useful in the study of the immune response to human blood group antigens.
The BEST Working Party of the International Society of Blood Transfusion: an international effort to improve the quality of blood components
Authors: Sirchia G.
Published: March 2003

BACKGROUND: The Biomedical Excellence for Safer Transfusion (BEST) Working Party of the International Society of Blood Transfusion was formed with the aim of improving the quality and safety of blood components. An outline of the aims, working methodology, and results after the first 4 years of activity of the BEST Working Party is presented. RESULTS: Seven studies have been performed, evaluating techniques for counting residual white cells in white cell‐reduced red cells, platelet quality control, and cellular harvests in blood components prepared with different methods. Areas of possible interest in the near future include stem cells, platelet crossmatching, and virus inactivation of blood components. An important BEST feature is the sharing of information gathered through collections of large sets of data from routine settings, so as to produce a reliable picture of the “real world.” CONCLUSION: Such information could prove useful to those in charge of developing recommendations and guidelines.
Blood donors who are repeatedly reactive for hepatitis C virus on enzyme immunoassay and positive on recombinant immunoblot assay: evidence of failure to identify some risk factors
Authors: Moore S. B. , Kruger J. R. , Rakela J. , Vamvakas E. C. , Schimek C. , Germer J. J. , Persing D. H.
Published: March 2003

BACKGROUND: Despite the introduction of surrogate testing and subsequent antibody testing of donor blood, transmission of hepatitis C virus (HCV) still occurs. The institution from which this report originates is a medical center, and many of the blood donors have also been seen as patients at the institution. This provided an opportunity for comparison of donor questionnaire responses and medical history information and for correlation of those findings with HCV test results. STUDY DESIGN AND METHODS: HCV polymerase chain reaction (PCR) testing was performed on nine stored frozen sera from donors or former donors with previous positive results on HCV enzyme immunoassay (EIA) (first‐ or second‐generation) and recombinant immunoblot assay (RIBA) (first‐ or second‐generation). The medical histories were also reviewed for 22 of 23 such HCV EIA‐repeatedly reactive, RIBA‐positive donors and 88 randomly chosen HCV‐negative donors. The lifestyle information was compared with the donors' responses on the blood donation questionnaires. The data were then correlated with available clinical and laboratory evidence of HCV transmission to transfusion recipients. RESULTS: For eight donors, there were no recipients of their blood to be assessed. For 9 of the remaining 15 donors, there were recipients who were tested for HCV. Recipients of blood from 5 of these 9 donors were repeatedly reactive for HCV; while recipients of blood from 4 donors were not. Donor PCR positivity correlated with apparent transmission (p = 0.047). Twelve of 20 HCV EIA‐positive donors for whom history and questionnaires were available for comparison had at least one suggestive lifestyle or established risk factor in their medical records, while none of 88 HCV‐negative controls did (p < 0.0001). The data did not indicate that paid donation correlated with failure to disclose these factors. CONCLUSION: Despite more explicit and intrusive donor questioning, it is still not possible to identify all possible risk factors at donations, though many donors who do not disclose all their risk factors are eliminated from the pool by the increasingly sensitive donor tests. As long as tests are not completely foolproof, workers must be vigilant regarding the ability to elicit complete information on a donor's risk. Further study is required to determine the best way(s) to do so.
Red cell alloimmunization after a bone allograft
Authors: Cheek Robert F. , Harmon James V. , Stowell Christopher P.
Published: March 2003

BACKGROUND: Alloimmunization to blood group antigens other than ABH after bone allografting is uncommon. To date, examples of alloimmunization to blood group antigens other than ABH have been limited to the Rh system. CASE REPORT: A 20‐year‐old woman received an allogeneic bone graft following resection of an osteosarcoma. The patient had received no blood components and denied any pregnancies. Alloimmunization was detected and alloantibodies were identified by standard serologic techniques. Fourteen months after the bone graft, anti‐Fya and anti‐Jkb were identified in her serum. CONCLUSION: Alloimmunization to blood group antigens other than ABO and Rh may follow bone allografting.
ISO 9000 quality standards: a model for blood banking?
Authors: Nevalainen David E. , Lloyd Huw L.
Published: March 2003

The recent American Association of Blood Banks publications Quality Program and Quality Systems in the Blood Bank and Laboratory Environment, the FDA's draft guidelines, and recent changes in the GMP regulations all discuss the benefits of implementing quality systems in blood center and/or manufacturing operations. While the medical device GMPs in the United States have been rewritten to accommodate a quality system approach similar to ISO 9000, the Center for Biologics Evaluation and Research of the FDA is also beginning to make moves toward adopting “quality systems audits” as an inspection process rather than using the historical approach of record reviews. The approach is one of prevention of errors rather than detection after the fact (Tourault MA, oral communication, November 1994). The ISO 9000 series of standards is a quality system that has worldwide scope and can be applied in any industry or service. The use of such international standards in blood banking should raise the level of quality within an organization, among organizations on a regional level, within a country, and among nations on a worldwide basis. Whether an organization wishes to become registered to a voluntary standard or not, the use of such standards to become ISO 9000‐compliant would be a move in the right direction and would be a positive sign to the regulatory authorities and the public that blood banking is making a visible effort to implement world‐class quality systems in its operations. Implementation of quality system standards such as the ISO 9000 series will provide an organized approach for blood banks and blood bank testing operations. With the continued trend toward consolidation and mergers, resulting in larger operational units with more complexity, quality systems will become even more important as the industry moves into the future.(ABSTRACT TRUNCATED AT 250 WORDS)
Reduced frequency of HLA‐DQ6 in individuals with a positive direct antiglobulin test
Authors: Wang‐Rodriguez J. , Rearden A.
Published: March 2003

BACKGROUND: The association between HLA class II antigens and the production of red cell autoantibodies was investigated in this study. STUDY DESIGN AND METHODS: Thirty‐one individuals with a positive direct antiglobulin test (DAT) and 85. DAT‐negative controls (cadaveric organ donors) were typed using a DNA‐based method for class II HLA typing, that is, the polymerase chain reaction with sequence‐specific primers. Amplified HLA alleles corresponding to the serologic specificities DR 1‐ 18 and DQ 1–9 were identified. RESULTS: Analysis of DR and DQ frequencies showed that HLA‐DQ6 was less frequent in DAT‐positive individuals (19% vs. 53% in the control population), with a p value of 0.0014 and a corrected p value of 0.059, and relative risk of 0.23 (95% Cl = 0.09‐0.58). The frequency of HLA‐DQ6 was higher in asymptomatic DAT‐positive blood donors (n = 8, 38% DQ6 positive, p = 0.48) than in DAT‐positive hospital and clinic patients (n = 23, 13% DQ6 positive, corrected p value = 0.030, relative risk = 0.13, 95% Cl = 0.04‐0.41), 96 percent of whom had evidence of clinical hemolysis. CONCLUSION: This study demonstrates that HLA‐DQ6 may have a negative association with a positive DAT result in patients with evidence for hemolysis, and may be a resistance antigen for clinically relevant red cell autoantibodies.
Alloimmunization after granulocyte transfusions
Authors: Stroncek D. F. , Leonard K. , Eiber G. , Malech H. L. , Gallin J. I. , Leitman S. F.
Published: March 2003

BACKGROUND: Although granulocyte transfusions are recommended for neutropenic patients with bacterial infections that are unresponsive to antibiotic therapy, the presence of white cell (WBC) antibodies in the recipient can render these transfusions ineffective. STUDY DESIGN AND METHODS: A 25‐year‐old man with chronic granulomatous disease experienced a pulmonary transfusion reaction while receiving granulocyte transfusions, and he was found to be immunized to neutrophil antigen NA2. A retrospective study of alloimmunization to HLA and neutrophil antigens in 18 patients with chronic granulomatous disease who had also received repeated granulocyte transfusions was then performed. Sera were tested in lymphocytotoxicity, granulocyte agglutination, granulocyte immunofluorescence, monoclonal antibody immonobilization of granulocyte antigen, and immunoprecipitation assays. RESULTS: After the granulocyte transfusions, sera from 14 of the 18 patients contained WBC antibodies. Seven sera samples reacted in the lymphocytotoxicity, granulocyte immunofluorescence, and granulocyte agglutination assays; seven reacted in the lymphocytotoxicity and granulocyte immunofluorescence assays but not the granulocyte agglutination assay, and four did not react. When the monoclonal antibody immobilization of granulocyte antigen assay was used, three sera samples reacted with Fc gamma receptor III, three with the 58‐ to 64‐kDa protein carrying the neutrophil antigen NB1, one with CD11a, and one with CD18. Antibodies from three patients immunoprecipitated a neutrophil protein of 60 kDa. Overall, antibodies to neutrophil antigens other than HLA could be detected in sera from eight patients. Transfusion reactions occurred in 11 of the 14 individuals with WBC antibodies and in none of the 4 without antibodies. Seven pulmonary reactions occurred in patients with WBC antibodies. The patients with WBC antibodies were given significantly more granulocyte concentrates (78 +/− 65 vs. 29 +/− 15 units, p < 0.05). CONCLUSION: Recipients of granulocyte transfusions often become alloimmunized. Screening for WBC antibodies periodically during transfusions, after adverse reactions, or before subsequent transfusions is indicated. If WBC antibodies are present, no further granulocyte transfusions should be given unless the granulocytes are collected from HLA‐ and/or neutrophil antigen‐ compatible donors.
The RoHar antigenic complex is associated with a limited number of D epitopes and alloanti‐D production: a study of three unrelated persons and their families
Authors: Beckers E.A.M. , Porcelijn L. , Ligthart P. , Vermey H. , von dem Borne A. E.G.Kr. , Overbeeke M. A.M. , van Rhenen D. J.
Published: March 2003

BACKGROUND: the RoHar antigenic complex has been characterized serologically by difficulties in D typing, weak e expression, lack of G antigen, presence of Rh33, a low‐frequency Rh antigen, and, more recently, a second low‐frequency antigen, FPTT. Allocation to one of the partial D catagories was not considered because of the unuaual reactions of RoHar cells and because anti‐D production was not observed in RoHar persons.

STUDY DESIGN and METHODS: Three unrelated RoHar donors and their families were studied in detail with special emphasis on D epitope mapping, e and G typing, and screening for antibodies.

RESULTS: Only D epitopes 5 and 6/7 were demonstrable, and D epitopes 1, 2, 3, 4, 8, and 9 seem to be absent in the RoHar complex. In one individual, the presence of alloanti‐D with limited specificity, not reacting with RoHar red cells of other individuals, was found 6 months after a second D+ pregnancy.

CONCLUSION: The finding of alloanti‐D in an RoHar r person supports the concept that the D characteristic of this phenotype is a partial D antigen, which is consistent with the presence of the limited number of D epitopes found in epitope mapping. As has been suggested for other partial D antigens, RoHar individuals should be regarded as D‐ for the receipt of blood, and pregnant RoHar women who have had D+ pregnancies should receive anti‐D prophylaxis.
Treatment of chronic autoimmune thrombocytopenic purpura with monoclonal anti‐D
Authors: Godeau B. , Oksenhendler E. , Brossard Y. , Bartholeyns J. , Leaute J.‐B. , Duedari N. , Schaeffer A. , Bierling P.
Published: March 2003

Background: The platelet count increases transiently after treatment with polyclonal anti‐D in about 50 percent of D+ patients with autoimmune thrombocytopenic purpura (AITP). The effect is usually attributed to macrophage Fc‐receptor blockade by antibody‐coated red cells. As polyclonal anti‐D is in limited supply, prospective testing was performed on a monoclonal anti‐D (MoAb D) in such patients.

Study Design and Methods: Seven D+ patients with chronic AITP received MoAb D intravenously at doses of 47 to 95 μg per kg of body weight. Response was assessed by studying platelet count increment. Hemolysis and red cell‐bound MoAb D were measured before and after MoAb D administration.

Results: MoAb D red cell binding was demonstrated in all patients at a ratio higher than that observed in AITP patients successfully treated with polyclonal anti‐D. However, little or no platelet count increment was observed in six patients, while a transient response was observed in only one (platelet count 97 × 109/L before MoAb D infusion and 163 × 109/L 4 days later). Furthermore, because five patients showed signs of hemolysis and two became anemic, higher doses of MoAb D should be used only with caution in patients with AITP.

Conclusion: The MoAb D used in this study cannot be proposed as an alternative treatment for patients with AITP.
Development and evaluation of a shipping system for platelet components
Authors: George V.M. , Pringle T.C. , Kline L. , Friedman L.I.
Published: March 2003

Background: Platelet concentrates and apheresis platelets must be maintained at a temperature as close as possible to 20 to 24°C during transport. To improve temperature control, ensure component quality, and meet handling and freight carrier needs, a new insulated shipping container system was developed and evaluated.

Study Design and Methods: Molded polyurethane‐insulated shipping containers were loaded with different payloads of simulated platelet components, with or without gel‐based temperature stabilizing packs (TSPs). The containers were subjected to constant ambient temperature of 37, 4 or ‐10°C. Payload temperatures were continuously monitored, in situ, for 24 hours.

Results: Temperature data are reported as the mean number of hours needed for components to warm or cool by 1 degree C. The temperature of payloads exposed to a constant 37°C ambient temperature increased by 1 degree C in 2.5 to 3.8 hours when no TSPs were included in the shipment and in 6.1 to 6.9 hours when TSPs were used. Exposure to a constant 4°C ambient temperature resulted in a 1 degree C temperature decrease in 1.8 to 3.4 hours without TSPs and in 4.6 to 5.6 hours with TSPs. At a ‐10°C ambient temperature, there was a 1 degree C drop within 1.0 to 1.6 hours without TSPs and within 2.7 to 2.9 hours with TSPs.

Conclusion: The container and packing methods described moderate the rate of change in the temperature of platelet components during their exposure to challenging ambient conditions. The use of TSPs substantially improves the performance of the system. In addition, the system meets freight carrier requirements and is easy to use, environmentally friendly, and durable.
Use of a directly conjugated monoclonal anti‐D (BRAD‐3) for quantification of fetomaternal hemorrhage by flow cytometry
Authors: Lloyd‐Evans P. , Kumpel B. M. , Bromelow I. , Austin E. , Taylor E.
Published: March 2003

BACKGROUND: Determination of the volume of fetal D‐positive cells in the circulation of D‐negative women after delivery is carried out to determine whether additional prophylactic anti‐D should be given to the mother. Although the Kleihauer‐Betke test is still widely used to calculate the fetomaternal hemorrhage, increasing use is being made of flow cytometry.

STUDY DESIGN AND METHODS: A conjugated monoclonal anti‐ D was prepared by labeling purified BRAD‐3 (IgG3) with fluorescein isothiocyanate (FITC‐BRAD‐3). This reagent was used to label D‐positive red cells by a one‐step procedure: 5 microL of washed cells were incubated with 50 microL of FITC‐BRAD‐3 (50 micrograms/mL) at 37 degrees C for 30 minutes; then the cells were washed and 500,000 events were analyzed by flow cytometry.

RESULTS: The FITC‐BRAD‐3 reagent effectively labeled D‐positive cells. The percentage of D‐positive cells in mixtures containing more than 0.04 percent D‐positive cells in D‐negative cells was accurately determined by using this reagent and flow cytometry. Although the Kleihauer‐Betke test was more accurate than this one‐step flow cytometric method at quantifying fetomaternal hemorrhage of < 1 mL, the flow cytometric method was more accurate in the 1‐ to 7‐mL fetomaternal hemorrhage range of 1 to 7 mL (whole‐blood equivalents). Analysis of 175 clinical samples for fetomaternal hemorrhage gave consistent quantification results with the three methods used: the Kleihauer‐Betke test, flow cytometry with FITC‐BRAD‐ 3, and flow cytometry with polyclonal anti‐D followed by FITC‐anti‐IgG.

CONCLUSION: Labeling of samples with FITC‐BRAD‐3 was simple and rapid. By flow cytometric analysis, good separation of D‐positive from D‐ negative cells was obtained, and fetomaternal hemorrhage of > 1 mL was quantified accurately.
Assessment of blood donor privacy during health history interviews
Authors: Kline L. , Friedman L. I. , Dempesy D. , Rabinowitz W. M. , Popovsky M. A.
Published: March 2003

BACKGROUND: Given the personal nature of health history interviews, it is important to provide donors with both visual and auditory privacy. Privacy is affected by variables such as background noise, the use of visual screens, and the loudness of the donor's voice.

STUDY DESIGN AND METHODS: In Phase I of this study, an interview station and waiting area were simulated. To measure auditory privacy, a speech intelligibility test was given to subjects with and without the use of a free‐standing privacy screen and masking noise device. Phase II was a field trial designed to evaluate screens and masking noise. Background noise was measured during each blood collection operation, and donors completed a survey.

RESULTS: In Phase I, speech intelligibility test scores ranged from 78 to 5.1 percent, depending on the type of visual screen and the number of masking noise devices used. In Phase II, with the use of screens, 94 percent of donors rated visual privacy as “good to excellent,” compared with 74 percent who did so when no screens were used. At many blood drives, the background noise level exceeded the level of the masking noise.

CONCLUSION: The use of visual screens increases donors' perception of visual privacy. The use of masking noise is effective only when the health history interview is conducted under conditions of low background noise levels.
Characterization of the hybrid RHD gene leading to the partial D category IIIc phenotype
Authors: Beckers E. A. M. , Faas B. H. W. , Ligthart P. , Simsek S. , Overbeeke M. A. M. , von dem Borne A. E. G.Kr. , van Rhenen D. J. , van der Schoot C. E.
Published: March 2003

BACKGROUND: A D‐positive white woman was found to have produced alloanti‐D leading to hemolytic disease of the newborn in her third D‐ positive child. The maternal D was identified as the partial D category IIIc antigen (DIIIc). The molecular basis of this phenotype was studied. STUDY DESIGN AND METHODS: The proposita and her relatives were phenotyped for Rh system antigens with standard reagents. D(IIIc) typing of D‐positive red cells was done with serum that contained anti‐ D from the proposita. Southern blot analysis and RHD‐specific polymerase chain reactions were performed with genomic DNA. Rh transcripts were cloned and sequenced. RESULTS: Six relatives of the proposita were found to express the DIIIc phenotype, which traveled with Ce. The DIIIc phenotype was inherited in a Mendelian fashion. Southern blot analysis showed an identical digestion pattern in D(IIIc) individuals and in DD controls. Three different Rh transcripts were found. Two Rh transcripts were derived from RHCE (RHce and RHCe). The RHD‐derived Rh transcript was the same as that of the published RHD sequence, apart from exon 3, which appeared to be exon 3 of RHCE. At the genomic level, RHD exon 3 was missing in all individuals expressing D(IIIc). CONCLUSION: This study shows the characteristics of a new hybrid D‐CE‐D allele encoding D(IIIc). It may be concluded that exon 3 of RHD is not involved in the formation of any of the D epitopes known at present, but rather encodes a new D epitope or D epitopes, as yet undefined by monoclonal anti‐D reagents.
Clinical toxicity and laboratory effects of granulocyte‐colony‐ stimulating factor (filgrastim) mobilization and blood stem cell apheresis from normal donors, and analysis of charges for the procedures
Authors: Anderlini P. , Przepiorka D. , Seong D. , Miller P. , Sundberg J. , Lichtiger B. , Norfleet F. , Chan K.‐W , Champlin R. , Korbling M.
Published: March 2003

BACKGROUND: Apheresis of granulocyte‐colony‐stimulating factor (filgrastim)‐mobilized blood stem cells from normal donors is now being used in place of a marrow harvest in transplantation. How the adverse effects of and charges for this procedure compare with those of the standard marrow harvest is not known.

STUDY DESIGN AND METHODS: Forty consecutive normal subjects who received filgrastim 96 micrograms/kg) subcutaneously twice daily for 4 to 6 days in preparation for apheresis were monitored prospectively by clinical and laboratory evaluation.

RESULTS: Sixty‐two percent of the subjects required oral analgesics. None discontinued filgrastim prematurely. Bone pain (82%), headache (70%), fatigue (20%), and nausea (10%) were reported. Filgrastim caused a mean eightfold increase in neutrophil counts, a mean twofold increase in lymphocyte counts, a mean twofold rise in alkaline phosphatase and lactate dehydrogenase levels, and minor changes in serum potassium, magnesium, and uric acid. Adverse events and laboratory effects resolved within 7 days after apheresis. No apheresis stem cell donor required transfusion or hospitalization, and only one required an additional clinic visit after completion of apheresis. By comparison, a retrospective analysis of 33 normal marrow donors demonstrated that all received transfusion(s), 3 were hospitalized, 3 required additional clinic visits after the marrow harvest. The median total charges related to the two procedures were comparable (p = 0.43), although the charges were significantly lower for donors requiring only one apheresis procedure (p = 0.002).

CONCLUSION: Filgrastim mobilization and apheresis of blood stem cells constitute a safe, well‐tolerated, and comparable or less expensive alternative to the traditional marrow harvest.