bluuebunny/crossref_metadata_2025 huggingface dataset:
Serologic characteristics of H‐deficient phenotypes among Chinese in Hong Kong
Authors: Mak K. H. , Lubenko A. , Greenwell P. , Voak D. , Yan K. F. , Poole J.
Published: March 2003

BACKGROUND: The occasional presence of H‐deficient red cells among both referred and donor blood samples prompted the mass screening of donated blood in Hong Kong for H‐deficient phenotypes; 96 percent of the donors tested are Chinese from the southern province of Kwongtung. STUDY DESIGN AND METHODS: Donor blood was screened for H‐deficient red cells with the use of Ulex europaeus. Lewis phenotyping was carried out on all H‐deficient individuals, and saliva testing was performed on most such individuals. The thermal amplitude and potency of their anti‐H and anti‐HI in the serum were also estimated. RESULTS: Between 1984 and 1993, 28 H‐deficient blood donors were identified; 16 H‐deficient patient samples were also identified, and family studies revealed an additional 7 H‐deficient subjects. The H‐deficient red cells did not react with anti‐H lectin, the levels of ABH substances in saliva were normal or near‐normal, normal levels of A or B transferase were found in plasma, minute quantities of A or B (in persons who were genetically group A or B) were detected on the red cells, and anti‐H or anti‐HI was detected in the serum (about 66.7% of which reacted at 37 degrees C). Atypical anti‐A or anti‐B was demonstrated in 81.8 percent of the cases. CONCLUSION: The H‐deficient phenotype among the Hong Kong Chinese seems to represent a homogeneous group. Despite the presence of normal quantities of ABH substance in the saliva, anti‐H or anti‐HI that was active at 37 degrees C was detected in most cases. The incidence of the H‐deficient phenotype was 1 in 15,620.
Kinetics of peripheral blood mononuclear cell mobilization with chemotherapy and/or granulocyte‐colony‐stimulating factor: implications for yield of hematopoietic progenitor cell collections
Authors: Webb I. J. , Eickhoff C. E. , Elias A. D. , Ayash L. J. , Wheeler C. A. , Schwartz G. N. , Demetri G. D. , Anderson K. C.
Published: March 2003

BACKGROUND: Peripheral blood progenitor cells (PBPCs) are commonly collected and used to reconstitute hematopoiesis after high‐dose chemotherapy. However, strategies for optimal collection and assessment of leukapheresis components are not standardized.

STUDY DESIGN and METHODS: Hematopoietic progenitor cell assays were performed on 369 leukapheresis components collected from 95 patients who had received doxorubicin‐based chemotherapy and/or granulocyte‐colony‐stimulating factor (G‐CSF). Precollection patient hematologic values, leukapheresis collection values, component hematopoietic progenitor cell assays, and patient outcome measures were summarized. The kinetics of mononuclear cell (MNC) and PBPC mobilization were assessed among four patient groups.

RESULTS: Patient group was a significant predictor of the peripheral blood MNC count on the day of collection (p<0.0001), and that value was a significant predictor of granulocyte‐macrophage– colony‐forming unit (CFU‐GM) yield (p<0.0001). This relationship between the peripheral blood MNC count on the day of collection and CFU‐ GM yield differed according to patient group (p<0.0001). CFU‐GM made up a larger fraction of peripheral blood MNCs collected from patients who received chemotherapy plus G‐CSF than collected from those who received G‐CSF alone. Moreover, the peripheral blood MNC count and the corresponding CFU‐GM yield increased significantly on consecutive days of collection in patient groups receiving chemotherapy and G‐CSF but were unchanged or decreased in patients receiving G‐CSF alone.

CONCLUSION: The relationship between peripheral blood MNC count and leukapheresis component CFU‐GM yield differed significantly between patients who received chemotherapy and G‐CSF and those who received G‐ CSF alone for the mobilization of PBPCs. Patient peripheral blood MNC count and component CFU‐GM yield are useful for both assessing and suggesting revisions to PBPC mobilization and collection strategies.
Determinants of red cell, platelet, plasma, and cryoprecipitate transfusions during coronary artery bypass graft surgery: the Collaborative Hospital Transfusion Study
Authors: Surgenor D. M. , Churchill W. H. , Wallace E. L. , Rizzo R. J. , Chapman R. H. , McGurk S. , Bertholf M. F. , Goodnough L. T. , Kao K. J. , Koerner T. A.W. , Olson J. D. , Woodson R. D.
Published: March 2003

BACKGROUND: Very little is known about the determinants of blood transfusions in patients undergoing coronary artery bypass graft surgery. STUDY DESIGN AND METHODS: To identify factors that influenced the transfusion of red cells, platelets, plasma, and cryoprecipitate, statistical methods were used to study 2476 consecutive diagnosis‐ related group 106 and 107 patients in five teaching hospitals who underwent coronary artery bypass surgery between January 1, 1992, and June 30, 1993. RESULTS: The likelihood of red cell transfusion was significantly associated with 10 preoperative factors: 1) admission hematocrit, 2) the patient's age, 3) the patient's gender, 4) previous coronary artery bypass surgery, 5) active tobacco use, 6) catheterization during the same admission, 7) coagulation defects, 8) insulin‐dependent diabetes with renal or circulatory manifestations, 9) first treatment of new episode of transmural myocardial infarction, and 10) severe clinical complications. Platelet and/or plasma transfusions were strongly associated with the dose of red cells transfused. Transfusion requirements and other in‐hospital outcomes were associated with patient characteristics, surgical procedure (reoperation vs. primary procedure), and the conduits used for revascularization (venous graft only, venous and internal mammary artery graft, or internal mammary artery graft only). Blood resource use and donor exposures were evaluated with respect to the risk to patients of contracting hepatitis C virus and human immunodeficiency virus infections. CONCLUSION: The classification of coronary artery bypass graft patients on the basis of attributes known preoperatively and by conduits used yields subsets of patients with distinctly different transfusion requirements and in‐ hospital outcomes.
RHD/CE typing by polymerase chain reaction using sequence‐specific primers
Authors: Gassner C. , Schmarda A. , Kilga‐Nogler S. , Jenny‐Feldkircher B. , Rainer E. , Muller T. H. , Wagner F. F. , Flegel W. A. , Schonitzer D.
Published: March 2003

BACKGROUND: Current DNA‐based Rh system typing strategies may detect the two RH genes and their prevalent alleles, but they are known to fail sometimes, when rare RH alleles (e.g., D category phenotypes) are encountered. It is almost impossible to find a single DNA‐based method that can accommodate the great heterogeneity within the human Rh system. STUDY DESIGN AND METHODS: An easy‐to‐perform DNA‐based method for the detection of the two RH genes and their alleles, including variant RHD alleles, was developed. By the use of one RHD/C‐, seven RHD‐, and four RHCE‐specific polymerase chain reactions, all triggered to work at identical thermocycling conditions, the DNA of 77 blood donors carrying weak D and that of 200 random donors with common D phenotype was investigated. In addition, 77 selected samples of ccDee and rare Rh system phenotypes were examined. RESULTS: Among 77 samples of weak D, one Rh33 and six DVI categories were detected, one of which showed new RHD‐specific nucleotide patterns. In DFR and CCee samples, novel variant RHD alleles were found. RHD DNA types of 200 random donors were found to be concordant with their D phenotype. For RHE and RHe genotyping, a full correlation with serologic phenotypes was found. Our method for genotyping RHC and RHc failed in some cases, because of an already published RHc allelic variation, which we have called RHc(cyt48). An estimate of the frequency of this RHc(cyt48) allele in a white population was made. CONCLUSION: The presented exon‐scanning RHD/CE polymerase chain reaction using sequence‐specific primers complements current DNA‐based Rh system typing strategies and is superior in the detection of variant RHD alleles.
Complement activation in vitro by the red cell substitute, liposome‐ encapsulated hemoglobin: mechanism of activation and inhibition by soluble complement receptor type 1
Authors: Szebeni J. , Wassef N.M. , Hartman K.R. , Rudolph A.S. , Alving C.R.
Published: March 2003

BACKGROUND: Liposome‐encapsulated hemoglobin (LEH) has been developed as an emergency blood substitute, yet its effect on human complement has never been explored. Considering that complement activation is a major pathogenic factor in the respiratory distress syndrome that often develops in trauma and shock, LEH‐induced complement activation may be a critical safety issue. STUDY DESIGN AND METHODS: Various LEH and corresponding empty liposomes were incubated with normal human sera, and various markers of complement activation (serum levels of C4d, Bb, SC5b‐9, and CH50; C5a‐induced granulocyte aggregation; membrane deposition of C3b) were measured. Incubations were also performed in the presence of (ethylene‐bis[oxyethylenenitrilo]tetraacetic acid) (EGTA) and Mg++ (EGTA/Mg++) and soluble complement receptor type 1. RESULTS: LEH and liposomes activated human complement, as indicated by significant changes in one or more markers. The effect was primarily due to the presence of the phospholipid vehicle; small, unilamellar, highly homodispersed vesicles induced the greatest degree of complement activation. Complement activation was partially inhibited by EGTA/Mg++. The latter finding, together with the parallel increases in serum C4d and Bb, suggests activation of both the classical and alternative pathways. Soluble complement receptor type 1 (0.05‐20 micrograms/mL) efficiently inhibited all vesicle‐induced complement activation. CONCLUSION: Because of complement activation, the use of LEH for transfusion may require careful evaluation of safety. Soluble complement receptor type 1 may be useful as a prophylactic agent for complement activation‐related complications of liposome infusions.
Mobilization kinetics of CD34+/Thy‐1dim progenitor cells during recombinant human granulocyte‐colony‐stimulating factor administration in normal donors
Authors: Seong C. , Durett A. , Mirza N. , Huh Y. , Anderlini P. , Champlin R.
Published: March 2003

BACKGROUND: Allogeneic transplantation of granulocyte‐colony‐ stimulating factor (G‐CSF)‐mobilized peripheral blood progenitor cells (PBPCs) from normal related donors is effective in achieving engraftment with a relatively short period of posttransplantation aplasia. The optimal dose and composition of PBPC transplants are unknown. The CD34+/Thy‐1dim progenitor cell subset is enriched for putative stem cells. STUDY DESIGN AND METHODS: The kinetics of the primitive subpopulation were prospectively studied in nine normal donors receiving recombinant human G‐CSF (6 microg/kg) subcutaneously twice daily for 6 days for collection of PBPCs for allogeneic transplantation. RESULTS: The concentration (mean +/− SD) of the circulating CD34+/Thy‐1dim subset increased from a baseline of 0.9 +/− 0.9 × 10(3) to 29.2 +/− 22.1 × 10(3) per mL on Day 4 and 38.0 +/− 29.8 × 10(3) per mL on Day 6. The level of CD34+/Thy‐1dim cells was closely correlated with the overall level of CD34+ cells. At baseline, CD34+/Thy‐1dim cells composed 21.1 percent of the total CD34+ cells, increasing to 36.3 percent at the peak of mobilization. CONCLUSION: CD34+/Thy‐1dim cells are optimally mobilized on Days 4 to 6 of recombinant human G‐CSF treatment.
Genotyping of RHD by multiplex polymerase chain reaction analysis of six RHD‐specific exons
Authors: Maaskant‐van Wijk P. A. , Faas B.H.W. , de Ruijter J.A.M. , Overbeeke M.A.M. , von dem Borne A.E.G.Kr. , van Rhenen D. J. , van der Schoot C. E.
Published: March 2003

BACKGROUND: Qualitative RHD variants are the result of the replacement of RHD exons by their RHCE counterparts or of point mutations in RHD causing amino acid substitutions. For RHD typing, the use of at least two RHD typing polymerase chain reaction (PCR) assays directed at different regions of RHD is advised to prevent discrepancies between phenotyping and genotyping results, but even then discrepancies occur. A multiplex RHD PCR based on amplification of six RHD‐specific exons in one reaction mixture is described. STUDY DESIGN AND METHODS: Six RHD‐ specific primer sets were designed to amplify RHD exons 3, 4, 5, 6, 7, and 9. DNA from 119 donors (87 D+, 14 D‐ and 18 with known D variants; whites and nonwhites) with known Rh phenotypes was analyzed. RESULTS: All six RHD‐specific exons from 85 D+ individuals were amplified, whereas none of the RHD exons from 13 D‐ individuals were amplified. Multiplex PCR analysis showed that the genotypes of two donors typed as D+ were DIVa and DVa. Red cell typing confirmed these findings. From all D variants tested (DIIIc, DIVa, DIVb, DVa, DVI, DDFR, DDBT) and from RoHar, RHD‐specific exons were amplified as expected from the proposed genotypes. CONCLUSION: The multiplex PCR assay is reliable in determining genotypes in people who have the D+ and partial D phenotypes as well as in discovering people with new D variants. Because the multiplex PCR is directed at six regions of RHD, the chance of discrepancies is markedly reduced. The entire analysis can be performed in one reaction mixture, which results in higher speed, higher accuracy, and the need for smaller samples. This technique might be of great value in prenatal RHD genotyping.
Distribution of HIV type 1 (HIV‐1) in blood components: detection and significance of high levels of HIV‐1 associated with platelets
Authors: Lee T. H. , Stromberg R. R. , Heitman J. W. , Sawyer L. , Hanson C. V. , Busch M. P.
Published: March 2003

BACKGROUND: Although inactivation of enveloped viruses transmitted by plasma derivatives has been successful, no methods for virus inactivation or removal have been established for platelet concentrates or red cell (RBC) components. Relatively little is known regarding the extent or significance of virus interactions with the cellular constituents in these components.

STUDY DESIGN AND METHODS: Units of whole blood were collected from six HIV type 1 (HIV‐1)‐positive, asymptomatic individuals and separated into peripheral blood mononuclear cells (PBMNCs), cell‐free plasma, white cell‐reduced platelet concentrate, and white cell‐reduced RBCs. DNA and RNA polymerase chain reaction and virus culture methods were used to study the compartmentalization of HIV‐1 immediately after component preparation and after storage.

RESULTS: As expected, HIV DNA and infectious virus were detected in fresh blood and in PBMNCs, and virion‐ associated RNA was detected in fresh plasma from all six donors. The levels of viral nucleic acids in these preparations remained relatively stable with 4 degrees C storage, whereas infectivity of PBMNCs was rapidly lost. Washed RBCs tested negative for HIV in all assays at all time points. Platelets retained high levels of HIV RNA (but not infectivity) after extensive washing, as well as after storage at 4 and 22 degrees C. High‐level platelet‐associated HIV‐1 was also demonstrated in samples collected during early seroconversion. Periseroconversion and postseroconversion levels of platelet‐associated HIV‐1 correlated with the level of plasma viremia and with the rate of progression to AIDS. Cell‐free virus from donor plasma and tissue culture fluid rapidly and firmly attached to platelets from noninfected donors. Infectivity of tissue culture virus bound to platelets was demonstrated in vitro.

CONCLUSION: Significant levels of HIV‐1 are associated with platelets during all stages of infection. Platelet‐ associated HIV could either mediate virus clearance or facilitate virus dissemination and expanded tropism. Finally, virus inactivation research must address virus associations with platelets.
Effects of nitric oxide on platelet activation during plateletpheresis and in vivo tracking of biotinylated platelets in humans
Authors: Stohlawetz P. , Horvath M. , Pernerstorfer T. , Nguyen H. , Vondrovec B. , Robisch A. , Eichler H.G. , Spitzauer S. , Jilma B.
Published: March 2003

BACKGROUND: The use of platelet transfusions has risen considerably over the last few years, which leads to the collection and transfusion of a greater number of donor plateletpheresis units. Plateletpheresis activates platelets in platelet concentrates, which determines the degree of the storage lesion subsequently observed.
CD36 deficiency is frequent and can cause platelet immunization in Africans
Authors: Lee K. , Godeau B. , Fromont P. , Plonquet A. , Debili N. , Bachir D. , Reviron D. , Gourin J. , Fernandez E. , Galactéros F. , Bierling P.
Published: March 2003

BACKGROUND: CD36 is expressed on several cell lineages. About 5 to 10 percent of Asians lack platelet membrane CD36 (pCD36), but the frequency of pCD36 deficiency in other ethnic groups is not known. Persons who are pCD36‐negative are apparently healthy but can develop CD36 isoimmunization.

STUDY DESIGN AND METHODS: The pCD36 phenotype was studied in 1885 subjects belonging either to a group of 1127 healthy French blood donors (almost all of whom were white Europeans) or to a group of 758 patients of known ethnic origin.

RESULTS: No pCD36‐negative persons were found among the blood donors. Only 1 of the 301 white European patients was pCD36‐negative. In contrast, 16 of the 206 sub‐Saharan Africans was pCD36‐negative, a proportion higher than that among that black Caribbeans (1/148, p<0.01). The frequency of pCD36‐negative patients was similar in blacks with and without sickle cell disease. Monocyte CD36 (mCD36) expression was studied in 15 of 22 pCD36‐negative individuals: it was <10 percent in 7 subjects (type I deficiency) and between 12 and 100 percent in 8 others (type II deficiency). Thirteen pCD36‐negative individuals had risk factors for immunization, and 4 had anti‐CD36. Some had a history resembling posttransfusion purpura (n = 2), platelet transfusion refractoriness (n = 1), and recurrent miscarriage (n = 1). No correlation was found between immunization and the amount of mCD36. Anti‐CD36 from an immunized type II‐deficient woman reacted with monocytes from normal controls but not with monocytes from type I‐ or type II‐deficient individuals, and thus it is postulated that mCD36 could be structurally different in normal and type II CD36‐deficient individuals.

CONCLUSION: CD36 deficiency is frequent in sub‐Saharan Africans; development of anti‐CD36 can lead to serious complications in multiply transfused patients, such as those with sicke cell disease.
Fresh, liquid‐preserved, and cryopreserved platelets: adhesive surface receptors and membrane procoagulant activity
Authors: Barnard M.R. , MacGregor H. , Ragno G. , Pivacek L.E. , Khuri S.F. , Michelson A.D. , Valeri C.R.
Published: March 2003

BACKGROUND: A study in humans showed that the transfusion of previously frozen human platelets after cardiopulmonary bypass, despite decreased survival, resulted in better hemostatic function than that of liquid‐preserved platelets stored at 22°C for 3 to 4 days.

STUDY DESIGN AND METHODS: In this study, fresh, 3‐ to 4‐day‐old liquid‐preserved, and cryopreserved human platelets were studied by the use of monoclonal antibodies directed against p‐selectin, glycoprotein (GP)Ib, activated GPIIb/IIIa, and coagulation factor V in a three‐color flow cytometric method.

RESULTS: The fresh and liquid‐preserved platelets had normal surface levels of GPIb, while the cryopreserved platelets were composed of distinct subpopulations of GPIb‐normal and GPIb‐reduced platelets. On the basis of the binding of factor V, both subpopulations of cryopreserved platelets exhibited greater surface binding of factor V than did fresh and liquid‐preserved platelets. Activated GPIIb/IIIa was elevated on GPIb‐normal platelets, but not on GPIb‐reduced platelets. Baboon platelets frozen by a procedure identical to that used to freeze human platelets also had GPIb‐normal and GPIb‐reduced subpopulations after the freezing‐thawing‐washing procedure. Autologous cryopreserved baboon platelets labeled with biotin‐X‐N‐hydroxysuccinimide showed a rapid removal of GPIb‐reduced platelets during the 5‐minute postinfusion period, whereas GPIb‐normal platelets had an in vivo recovery of 48 percent and a lifespan of slightly less than 6 days.

CONCLUSIONS: Improved in vivo function of cryopreserved platelets may be related to the rapid hemostatic effect of the GPIb‐reduced subpopulation secondary to increased binding of factor V and expression of p‐selectin.
Molecular analysis of secretor type α(1,2)‐fucosyltransferase gene mutations in the Chinese and Thai populations
Authors: Chang J.G. , Yang T.Y. , Liu T.C. , Lin T.P. , Hu C.J. , Kao M.C. , Wang N.M. , Tsai F.J. , Peng C.T. , Tsai C.H.
Published: March 2003

BACKGROUND: The human Lewis histo‐blood group system belongs to a family of structurally related oligosaccharides. The mutations of fucosyltransferase genes α(1,2)‐fucosyltransferase (FUT2 or Se) and α(1,3/1,4)‐fucosyltransferase (FUT3 or Le), are responsible for the polymorphism of Lewis blood group phenotypes. However, a population study of the FUT2 mutation in Chinese and Thais has not yet been done, and there is some controversy about the phenotypes of Le(a+b+) and Le(a+b‐).

STUDY DESIGN AND METHODS: One hundred twenty‐four Chinese and 70 Thais were phenotyped for Lea and Leb. DNA samples were studied by polymerase chain reaction and then by a restriction enzyme digestion method to distinguish wild‐type and six known mutations. Direct sequencing was done for controls and some uncertain cases.

RESULTS: A new mutation, C302T mutation, was found in 2 of 136 chromosomes in the Thai population; none were discovered in Chinese. The frequencies of the normal and six mutant alleles among Chinese and Thais, respectively, were as follows: 134 (54.0%) of 248 and 58 (41.4%) of 140 were wild‐type (Se); 0 of 248 and 2 of 140 (both 1.4%) had the G428A mutation; 120 (48.4%) of 248 and 75 (53.6%) of 140 had the A385T mutation; 2 (0.81%) of 248 and 0 of 140 had the C571T mutation; and 1 (0.4%) of 248 and 3 (2.2%) of 140 had the G849A mutation. Only 1 Chinese (0.4%) of 248 had the C628T mutation, and none had fusion gene mutation.

CONCLUSION: The FUT2 genes encoding for the phenotypes Le(a+b+) and Le(a+b‐) are the same. The function and character of the mutant enzyme may play an important role in the phenotype. The methods used in this study are clinically applicable in population studies of the FUT2 gene polymorphism to explore relationships among different ethnic groups and correlations between phenotype and genotype.
Trends in the incidence of delayed hemolytic and delayed serologic transfusion reactions
Authors: Pineda A.A. , Vamvakas E.C. , Gorden L.D. , Winters J.L. , Moore S.B.
Published: March 2003

BACKGROUND: An increasing incidence of delayed hemolytic and delayed serologic transfusion reactions (DHTRs/DSTRs) has been seen at the Mayo Clinic since 1978. Recently, the average length of stay (LOS) for inpatients and the average number of red cell transfusions per inpatient (TPI) decreased, and the albumin and papain technique for RBC antibody detection was replaced by a polyethylene glycol technique. These changes may have affected the incidence of DHTRs/DSTRs.

STUDY DESIGN AND METHODS: The diagnoses of DHTR and DSTR made at the Mayo Clinic from 1993 through 1998 were reviewed. These data were compared with previously published Mayo Clinic data from 1980 through 1992. The LOS for inpatients and the average TPI were also obtained from hospital data.

RESULTS: The incidence of DHTR/DSTR increased from 1 in 1899 in 1980 through 1992 to 1 in 1300 in the 1993 through 1998 period (p<0.05). Similarly, DSTR increased from 1 in 2990 in 1980 through 1992 to 1 in 1612 in the 1993 through 1998 period (p<0.05). The incidence of DHTR showed a trend toward decrease, from 1 in 5405 in 1980 through 1992 to 1 in 6715 in 1993 through 1998. No alloantibody specificities were statistically associated with DHTRs in 1993 through 1998, unlike in the 1980 through 1992 period. Moreover, the incidence of Jka antibodies increased in 1993 through 1998, while the incidence of other alloantibodies remained stable. Average LOS and TPI declined by 24.5 percent and 8.8 percent, respectively, between the two periods.

CONCLUSION: Recently, a trend toward a decrease in the incidence of DHTR and a significant increase in DSTRs has occurred at the Mayo Clinic. These changes are most likely due to a combination of factors, including a decrease in average LOS and the adoption of the polyethylene glycol antibody detection system.
Immunoglobulin administration to fetuses with anemia due to alloimmunization to D
Authors: Ulm B. , Kirchner L. , Svolba G. , Jilma B. , Deutinger J. , Bernaschek G. , Panzer S.
Published: March 2003

BACKGROUND: The purpose of this study was to examine fetal tolerance of high‐dose intravenous immunoglobulin (IVIG), given directly at the time of intravascular transfusion, and its effects on fetal hemolysis and pregnancy outcome in the setting of alloimmunization to D.

STUDY DESIGN AND METHODS: Thirteen consecutive D+ fetuses requiring transfusion for maternal alloimmunization received high‐dose IVIG (1.0 g/kg) and red cell transfusions. Twenty‐four previous, consecutive fetuses with maternal anti‐D served as controls. The schedules for subsequent transfusions were the same in the two groups.

RESULTS: High‐dose IVIG was well tolerated by all fetuses. In the IVIG group, daily decreases in hematocrit were smaller than those in controls after the second administration of IVIG (mean hematocrit decrease, 0.72 percent/day vs. 1.45 percent/day; p = 0.007). No significant difference was found in the total number of fetal transfusions, the gestational age at delivery, the duration of neonatal intensive care, the number of neonates requiring postnatal transfusion therapy, and perinatal mortality.

CONCLUSION: In this small pilot study, direct administration to fetuses of IVIG with red cell transfusions was well tolerated and appeared to have a beneficial effect on fetal hemolysis.
NOR polyagglutination and Sta glycophorin in one family: relation of NOR polyagglutination to terminal α‐galactose residues and abnormal glycolipids
Authors: Kuśnierz‐Alejska Grazyna , Duk Maria , Stony Jill R. , Reid Marion E. , Wiȩcek Barbara , Seyfried Halina , Lisowska Elwira
Published: March 2003

BACKGROUND: This report describes the characterization of polyagglutinable red cells (RBCs), identified in two generations of a Polish family.

CASE REPORT: Untreated and modified RBCs of the proposita (TS) were tested by serologic methods, using human sera, antibodies, lectins, and inhibitors of agglutination. Moreover, glycophorins were characterized by sodium docecyl sulfate‐polyacrylamide gel electrophoresis and Western blotting, and glycolipids were purified, fractionated by thin‐layer chromatography, and detected with Ricinus communis agglutinin I (RCA‐I, specific for galactose residues) and Griffonia simplicifolia IB4 lectin (GSL‐IB4, specific for Galα1–3Gal‐ structure). Some of the experiments were also performed on RBCs of members of TS's family.

RESULTS: Polyagglutination, found in four members of TS's family, was identified as the second case of an earlier described NOR polyagglutination. The polyagglutination was decreased by treating the RBCs with α‐galactosidase and was inhibited by a neutral glycolipid fraction from NOR+ RBCs. Detection of neutral glycolipids of TS's RBCs on the thin‐layer plate by RCA‐I and GSL‐IB4 revealed the presence of components that were not detectable in control RBCs. Moreover, Western blotting of RBC membranes from five family members with glycophorin monoclonal antibodies and agglutination assays with anti‐Sta and anti‐Dantu sera identified the presence of Sta glycophorin in four members of the family, two of whom were NOR+ and two NOR–.

CONCLUSION: Our results showed that two rare features of TS's RBCs, NOR polyagglutination and Sta glycophorin, are inherited independently, and that NOR+ RBCs contain neutral glycolipids with an abnormal oligosaccharide structure, most likely terminated with α‐galactosyl residues.
Alteration of red cell aggregability and shape during blood storage
Authors: Hovav Tavor , Yedgar Saul , Manny Noga , Barshtein Gregory
Published: March 2003

BACKGROUND: Storage of blood units (for 35–42 days, depending on the preservative solution) has been reported to induce changes (e.g., reduction of sialic acid level) in red cells that are expected to alter their aggregability.

STUDY DESIGN AND METHODS: The aggregability of stored red cells was monitored in their autologous plasma and compared to that obtained with washed cells in dextran‐containing buffer throughout the storage period. Red cell aggregability was determined by using a computerized image analyzer of cell flow properties.

RESULTS: Blood storage induced changes in red cells that are associated with continuous increase of their aggregability. At the same time, blood storage was associated with a reduction in the level of plasma fibrinogen, the major aggregating agent in plasma. Accordingly, the increased red cell aggregability was observed in red cells stored in dextran‐containing buffer, but not in red cells stored in autologous plasma.

CONCLUSION: Because blood transfusion is routinely given to patients with normal or high fibrinogen level, the transfusion of stored red cells has the potential to induce increased aggregation in vivo, depending on the storage period. This should be taken into account when blood transfusion is considered, particularly for patients with microcirculatory disorders.
Length of storage of transfused red cells and postoperative morbidity in patients undergoing coronary artery bypassgraft surgery
Authors: Vamvakas E.C. , Carven J.H.
Published: March 2003

BACKGROUND: The transfusion of old red cells (RBCs) may be associated with reduced delivery of oxygen to tissues and an increased risk of transfusion complications. The association of postoperative morbidity with the length of storage of perioperatively transfused RBCs was studied in 268 consecutive patients receiving a blood transfusion for coronary artery bypass graft surgery.

STUDY DESIGN AND METHODS: The postoperative length of hospitalization, the postoperative length of stay in the intensive care unit, and the length of endotracheal intubation after the day of the operation were used as surrogate measures of global postoperative morbidity. The length of storage of the oldest transfused RBC unit, the mean length of storage of the oldest and second oldest RBC units, and the mean length of storage of all RBC units transfused to each patient were tested for association with the three outcome variables. Multiple linear regression analysis and Cox proportional‐hazard analysis were used to adjust for the effects of confounding factors that pertained to each patient's severity of illness and the difficulty of each operation.

RESULTS: There were no significant associations after adjustment for the effects of confounding factors.

CONCLUSION: This study did not corroborate the previously reported association between transfusion of old RBCs and increased morbidity. However, there is surprisingly little research on the clinical outcomes of the transfusions of old RBCs, and this hypothesis should be investigated further.
Molecular basis of Cromer blood group antigens
Authors: Lublin D.M. , Kompelli S. , Storry J.R. , Reid M.E.
Published: March 2003

BACKGROUND: The Cromer blood group system consists of 10 antigens located on decay‐accelerating factor (DAF). Previous molecular genetic analysis has determined the basis for four of these antigens. The present study was undertaken to identify the mutations that determine the remaining antigens.

STUDY DESIGN AND METHOD: Existing or new data were used to localize each Cromer system antigen to a specific short consensus repeat (SCR) domain of DAF. The exon encoding that SCR domain was amplified by using the polymerase chain reaction (PCR) on genomic DNA obtained from individuals of that Cromer phenotype, and the DNA product was subjected to DNA sequence analysis.

RESULTS: The Tca/Tcc polymorphism is due to an R18P amino acid substitution in SCR1 of DAF. The Es(a+)/Es(a–) polymorphism is due to an I46N mutation in SCR1 of DAF. The WESb/WESa polymorphism is due to an L48R mutation in SCR1 of DAF. The UMC+/UMC– polymorphism is due to a T216M substitution in SCR4 of DAF.

CONCLUSIONS: With information from previous reports and the findings of this study, the molecular genetic basis of all known alleles of the Cromer blood group system has been elucidated. Single amino acid substitutions are responsible for 9 of the 10 antigens (all except the multiple‐epitope antigen IFC).
The utility of ≤3‐day‐old whole‐blood platelets in reducing the incidence of febrile nonhemolytic transfusion reactions
Authors: Kelley D.L. , Mangini J. , Lopez‐Plaza I. , Triulzi D.J.
Published: March 2003

BACKGROUND: Febrile nonhemolytic transfusion reactions (FNHTRs) to platelet transfusions have been linked to the presence of cytokines in supernatant plasma. Cytokine concentration is directly related to WBC content and storage time. This study evaluated the effect of limiting the storage time of random‐donor platelet concentrates on the FNHTR rate.

STUDY DESIGN AND METHODS: FNHTR rates were calculated retrospectively for single‐donor apheresis platelet (SDP) and pooled random‐donor platelet (PP) transfusions given during three consecutive 5‐month study periods (November 1995 to February 1997) to patients on a single hematology/oncology/bone marrow transplant unit. Transfusion practice policies were: Baseline Period, SDPs preferred; Study Period A, PPs preferred; and Study Period B, ≤3‐day‐old PPs preferred. FNHTR rates were calculated from physicians' interpretations of reported reactions and the total number of SDP and PP transfusions in each period. SDPs were collected on two cell separators. All platelet components were filtered at issue in the laboratory by WBC‐reduction filters.

RESULTS: FNHTR rates for PP transfusions were: baseline, 11.1 percent (3/27); Study Period A, 4.6 percent (22/481); and Study Period B, 1.1 percent (3/282). The rates for SDP transfusions were 0.15 percent (1/650), 0.75 percent (2/267), and 0.36 percent (1/273), respectively. The FNHTR rate for ≤3‐day‐old PPs was significantly less than the rate for older PPs (p = 0.0086 for Study Period A vs. Study Period B), and was not significantly different than that for SDPs (p = 0.33 for PPs vs. SDPs in Study Period B).

CONCLUSION: Limiting transfusion of PPs to those stored ≤3 days is an effective strategy in reducing the rate of FNHTR and results in an FNHTR rate comparable to that seen with SDPs.
HPC viability measurement: trypan blue versus acridine orange and propidium iodide
Authors: Mascotti K. , McCullough J. , Burger S.R.
Published: March 2003

BACKGROUND: A reliable, validated method for rapidly determining HPC viability is essential for clinical cell engineering.

STUDY DESIGN AND METHODS: A fluorometric cell viability assay using acridine orange and propidium iodide (AO/PI) was compared to the current standard, trypan blue (TB) exclusion. Viable cells stained with AO/PI fluoresce green under darkfield fluorescence microscopy, while nonviable cells fluoresce orange. Mixtures of fresh and heat‐killed bone marrow were prepared and used as viability standards for evaluation of both assays. The frequency of CFU‐GM was determined for each specimen.

RESULTS: Cell viability measured by AO/PI was extremely linear, with measured and predicted viability in agreement from 0 to 100 percent of the viable cells and a coefficient of regression (r2) of 0.9921. The predicted‐viability regression line fell within the 95% CI for AO/PI‐measured viability. The coefficient of regression for TB‐measured viability was 0.9584, with the predicted‐viability regression line almost entirely outside the 95% CI. TB overestimated the percentage of viable cells, particularly below the 50‐percent level. CFU‐GM frequency correlated better with cell viability measured by AO/PI (r2 = 0.979) than with that measured by TB (r2 = 0.930).

CONCLUSIONS: The AO/PI viability assay is a rapid, highly linear, functionally correlated assay that is superior to conventional viability measurement by TB exclusion.
Inactivation of WBCs in RBC suspensions by photoactive phenothiazine dyes: comparison of dimethylmethylene blue and MB
Authors: Skripchenko A.A. , Wagner S.J.
Published: March 2003

BACKGROUND: The transfusion of blood components containing WBCs can cause unwanted complications, which include virus transmission, transfusion‐associated GVHD, alloimmunization, febrile reactions, and immunomodulation. Phototreatment with 4 μM of dimethylmethylene blue (DMMB) and 13 J per cm2 of white light irradiation has previously been shown to be an effective way to inactivate different models of enveloped and nonenveloped viruses in RBC suspensions, with minimum damage to RBCs. The present study compares WBC photoinactivation in buffy coat after DMMB or MB phototreatment under virucidal conditions.

STUDY DESIGN AND METHODS: Buffy coat diluted to 30‐percent Hct was treated with the dye and white light. Isolated WBCs were assayed for cell proliferation and viability by an assay using a tetrazolium compound, limiting dilution analysis, DNA fragmentation, and flow cytometry assays.

RESULTS: DMMB and 2.5 J per cm2 of light phototreat‐ment can inactivate T cells to the limit of detection by limiting dilution analysis (>4.76 log reduction). No WBC proliferation activity was observed after DMMB and 3.8 J per cm2 of light. DNA degradation after DMMB phototreatment was light dependent. In addition, DMMB phototreatment induced apoptosis in WBCs. In contrast, MB phototreatment under virucidal conditions did not cause significant changes in the viability of WBCs. Neither DNA degradation nor signs of apoptosis were observed after MB phototreatment.

CONCLUSION: DMMB phototreatment inactivates T‐lymphocytes, the cells that cause GVHD.
RBC antibody persistence
Authors: Schonewille Henk , Haak Hans L. , Van Zijl Annette M.
Published: March 2003

BACKGROUND: A person exposed to foreign blood group antigens may produce antibodies. The persistence of antibodies varies among people and among antibodies. A study was performed to investigate the persistence of clinically significant RBC alloantibodies over a period of 20 years.

STUDY DESIGN AND METHODS: A retrospective examination was performed of all records of RBC antibodies in the transfusion laboratory computer database from 1978 through 1997. Records of patients who underwent at least one antibody investigation after an antibody had been detected were studied. The study included all antibodies against the Rh, K, Fy, Jk, and MNs blood group systems. An antibody was regarded as not persistent if, after previous detection, the screening or panel studies became negative for the antibody under study. Anti‐D due to RhIg administration was excluded.

RESULTS: An analysis was performed of 480 records consisting of 593 antibodies that fulfilled the criteria. Median antibody follow‐up was 10 months (range, 1‐240). In 137 patients, 153 (26%) antibodies became undetectable over the course of time. After initial negative screening investigations, 310 antibodies were formed. The antibodies that were still detectable had a median follow‐up of 7 months (range, 1‐193). A patient's age, sex, and antibody specificity were of no influence on the length of time that antibodies were detectable. Antibodies detected with a more sensitive screening technique were less persistent (p = 0.0002). For 28 patients, detection of antibodies was highly irregular.

CONCLUSIONS: About 25 percent of all antibodies became undetectable over the course of time. The antibody screening technique used, rather than the antibody specificity, affected these results. To prevent delayed hemolytic transfusion reactions, precise antibody documentation is of great importance.
The survival, function, and hemolysis of human RBCs stored at 4°C in additive solution (AS‐1, AS‐3, or AS‐5) for 42 days and then biochemically modified, frozen, thawed, washed, and stored at 4°C in sodium chloride and glucose solution for 24 hours
Authors: Valeri C. Robert , Pivacek Linda E. , Cassidy George P. , Ragno Gina
Published: March 2003

BACKGROUND: A study was done to assess the quality of RBCs stored at 4°C in AS‐1, AS‐3, or AS‐5 for 42 days before biochemical modification and freezing.

STUDY DESIGN AND METHODS: RBCs were stored at 4°C for 42 days in AS‐1, AS‐3, or AS‐5 and then biochemically modified with pyruvate, inosine, phosphate, and adenine solution (Rejuvesol), frozen with 40‐percent (wt/vol) glycerol, and stored at –80°C for at least 2 months. The RBCs were deglycerolized by the use of a cell washer (Haemonetics 115), and stored for 24 hours at 4°C in a 0.9‐percent sodium chloride and 0.2‐percent glucose solution before the autologous transfusion.

RESULTS: The mean freeze‐thaw‐wash recovery process produced RBC recovery values of 85 percent, with the mean 24‐hour posttransfusion survival at 75 percent, and the mean index of therapeutic effectiveness at 64 percent for the RBCs stored at 4°C in AS‐1, AS‐3, or AS‐5 for 42 days before biochemical modification and freezing. All the units exhibited normal or slightly higher than normal 2,3 DPG levels after deglycerolization and postwash storage at 4°C for 24 hours.

CONCLUSION: RBCs stored in AS‐1, AS‐3, or AS‐5 at 4°C for 42 days and then biochemically modified with pyruvate, inosine, phosphate, and adenine and glycerolized, frozen, washed, and stored at 4°C for 24 hours before autologous transfusion had acceptable in vitro and in vivo measurements.
Isolation of an IgG anti‐B from a human Fab‐phage display library
Authors: Chang Tylis Y. , Siegel Don L.
Published: March 2003

BACKGROUND: ABO incompatibility is a common cause for mild hemolysis in the newborn, ranging from 1 in 30 to 1 in 150 births. Fortunately, hemolysis requiring transfusion is rare and restricted to blood group O mothers, because blood group A and B individuals make poor IgG anti‐B and anti‐A responses. No human IgG ABO antibody sequences have been reported, in part because of the difficulty in obtaining human IgG hybridomas. Phage‐display technology may be able to circumvent these difficulties, but its application to carbohydrate antigens is poorly studied.

STUDY DESIGN AND METHODS: A human IgG1 phage‐display Fab library was constructed from splenocytes derived from a nonhyperimmunized blood group O person, and panned against group B RBCs.

RESULTS: After five rounds of panning, essentially all phage bound to group B RBCs. Nucleotide sequence analysis of a single monoclonal IgG1λ phage, FB5.7, revealed a highly mutated VH4 family heavy chain, and a nearly germline VL7 family λ light chain. The Fab agglutinated group B, but not group A, random‐donor RBCs. However, group B ELISA reactivity could be inhibited by soluble B‐trisaccharide, soluble A‐trisaccharide, galactose, and N‐acetyl galactosamine. Similarly, galactose and N‐acetyl galactosamine were able to inhibit group B RBC agglutination.

CONCLUSION: FB5.7 is the first human IgG ABO MoAb described. Alhough it behaves serologically like a group B‐specific antibody, it demonstrates interaction with both the A and B epitopes. Phage‐display technology can be used to better define the relationship between antibody genotype and phenotype in anti‐carbohydrate responses in nonhyperimmunized hosts, and thus to improve our understanding of the composition of the antibody repertoire.
PCR screening for common weak D types shows different distributions in three Central European populations
Authors: Müller Thomas H. , Wagner Franz F. , Trockenbacher Andreas , Eicher Nicole I. , Flegel Willy A. , Schönitzer Diether , Schunter Friedrich , Gassner Christoph
Published: March 2003

BACKGROUND: DNA sequencing showed RHD mutations for all weak D phenotypes investigated in a study from Southwestern Germany. Molecular classification of weak D offers a more reliable basis than serotyping and is relevant for optimal D transfusion strategies.

STUDY DESIGN AND METHODS: Sequence‐specific primers were designed to detect weak D types 1 to 5 and the partial D phenotype HMi in a modular set for conventional PCR analysis. Alternatively, all reactions were multiplexed into a single tube, and the products were identified after automated capillary electrophoresis by their size and fluorescence. Weak D phenotype samples from 436 donors in the Tyrol (Austria) and Northern Germany were investigated by PCR.

RESULTS: More than 90 percent of the weak D types identified by PCR represented type 1, 2, or 3. The distribution among the common types varied between the Tyrol and Northern Germany (p<0.0001). Three new RHD alleles were identified.

CONCLUSION: A PCR method of detecting the common weak D types was validated. This PCR system introduces a simple and rapid tool for routine DNA typing of weak D samples. The results confirmed that all weak D phenotype samples identified by current serologic criteria carry altered D proteins.
Umbilical cord blood progeny cells that retaina CD34+ phenotype after ex vivo expansion have less engraftment potential than unexpanded CD34+ cells
Authors: Xu Ruiling , Reems Jo Anna
Published: March 2003

BACKGROUND: Because of the limitation of cell numbers associated with cord blood harvests, there is a need to determine the efficacy of using ex vivo‐expanded cord blood cells in a transplantation setting. In this study, limiting‐dilution analysis was used in nonobese diabetic mice with severe combined immunodeficiency (NOD/SCID) to compare the engraftment potential of progeny cells expressing the CD34+ phenotype after expansion with that of uncultured CD34+ cells.

STUDY DESIGN AND METHODS: Cord blood CD34+ cells were cultured in Iscove's modified Dulbecco medium supplemented with 10‐percent fetal calf serum (FCS) and IL‐6, SCF, megakaryocyte growth and development factor, and Flt3 ligand. The resulting ex vivo‐expanded products were assessed for total numbers of nucleated cells, CD34+ cells, and CFUs and long‐term culture‐initiating cell activity. The engraftment potentials of cultured progeny CD34+ cells and uncultured CD34+ cells were determined by using NOD/SCID mice.

RESULTS: After 14 days of culture, total nucleated cell counts increased over input values by 180 ± 59‐fold, CD34+ cell numbers by 44 ± 13‐fold, CFU activity by 23 ± 5‐fold, and long‐term culture‐initiating cell activity by 20 ± 6‐fold (mean ± SD; n = 6). The frequency of SCID‐repopulating cells (SRC) in mice transplanted with uncultured products was 1 per 20,000 CD34+ cells (95% CI, 1:10,000‐1:38,000) and that in mice receiving ex vivo‐expanded products was 1 per 418,000 progeny CD34+ cells (95% CI, 1:158,000‐1:1,100,000). Taken together, these data indicated that, after 2 weeks of culture, there was a modest twofold increase in the total number of SRCs. However, the levels of human CD45 cell engraftment in NOD/SCID recipients of progeny CD34+ cells were significantly lower than those in mice receiving equivalent numbers of uncultured CD34+ cells (p<0.05).

CONCLUSION: Umbilical cord blood progeny cells retaining a CD34+ phenotype after ex vivo expansion have less engraftment potential than do unexpanded CD34+ cells.