bluuebunny/crossref_metadata_2025_split huggingface dataset:
Soluble vascular endothelial growth factor in various blood transfusion components
Authors: Nielsen H.J. , Werther K. , Mynster T. , Brünner N. 
Published: March 2003

BACKGROUND: Blood transfusion may reduce survival after curative surgery for solid tumors. This may be related to extracellular content of cancer growth factors present in transfusion components. Vascular endothelial growth factor (VEGF) is a potent stimulator of angiogenesis in solid tumors. The potential content of VEGF in various blood components for transfusion was evaluated.STUDY DESIGN AND METHODS: Soluble VEGF (sVEGF, isotype 165) was determined by an enzyme‐linked immunosorbent assay (EIA) in serum and plasma samples and in lysed cells from healthy volunteers. Subsequently, total content of sVEGF was determined in nonfiltered and prestorage white cell‐reduced whole blood (WB), buffy coat‐depleted saline‐adenine‐glucose‐mannitol (SAGM) blood, platelet‐rich plasma (PRP), and buffy coat‐derived platelet (BCP) pools obtained from volunteer, healthy blood donors. As a control, total content of platelet‐derived soluble plasminogen activator inhibitor type 1 (sPAI‐1) was determined by an EIA in the same samples. Finally, the extracellular accumulation of sVEGF was determined in nonfiltered WB and SAGM blood during storage for 35 days and in BCP pools during storage for 7 days.RESULTS: In the healthy volunteers, median total sVEGF content was 97 (range, 20‐303) pg per mL in serum and 19 (13‐37) pg per mL in plasma (n = 12, p<0.002) and 445 (280‐990) pg per mL in lysed cells. Median total sPAI‐1 content was 94 (64‐127) ng per mL in serum, 8 (6‐11) ng per mL in citrated plasma, and 95 (78‐123) ng per mL in lysed cells. In SAGM blood, the median total sVEGF content was 25.3 (3.3‐48.4) ng per unit in nonfiltered units and undetectable in white cell‐reduced units. Median total sVEGF content was 29.2 (24.8‐124.9) ng per unit in nonfiltered PRP and 28.7 (24.5‐118.6) ng per unit in white cell‐reduced PRP. The sVEGF accumulated significantly in WB, SAGM blood, and BCP pools, depending on the storage time.CONCLUSION: The sVEGF (isotype 165) appears to be present in various blood transfusion components, depending on storage time.
Genomic analysis of ABO chimeras and mosaicsusing hematopoietic colony‐derived DNA
Authors: Anan K. , Suzuki H. , Iwasaki M. , Kobayashi K. 
Published: March 2003

BACKGROUND: While there are many case reports dealing with ABO mosaicism and chimerism, there have been few attempts to determine the patient's genotype.STUDY DESIGN AND METHODS: Peripheral blood and buccal mucosa were obtained from three persons with ABO mosaicism or chimerism. DNA extracted from hematopoietic progenitor cell‐derived colonies and from peripheral blood and buccal mucosa were analyzed by polymerase chain reaction‐restriction fragment length polymorphism methods. In addition, analyses of short tandem repeat markers were carried out.RESULTS: Hematopoietic progenitor cell‐derived DNA analysis revealed that, in two of the three persons there were 2 apparently distinct progenitor cell lineages whose percentages were close to those in the peripheral blood of the patients, as analyzed by flow cytometry; the exception was Subject 3, who had myelodysplastic syndrome (MDS). Short tandem repeat analysis showed that the former two subjects had two pairs of ABO alleles and the latter subject, with MDS, had loss of heterozygosity in some colony‐derived DNA as well as blood DNA.CONCLUSION: The subjects without MDS had two distinct hematopoietic cell lineages that led to their ABO chimeric status. The subject with MDS was assumed to have an ABO mosaicism caused by the somatic deletion of the ABO gene in the hematopoietic progenitor cells.
Site‐directed mutagenesis of the human D antigen: definition of D epitopes on the sixth external domain of the D protein expressed on K562 cells
Authors: Liu Wendy , Smythe Jonathan S. , Scott Marion L. , Jones Jeffrey W. , Voak Douglas , Avent Neil D. 
Published: March 2003

BACKGROUND: The antigens of the human Rh system are of great clinical significance in transfusion medicine and pregnancy. Of the Rh system antigens, D is clinically the most important, being one of the most immunogenic structures arising from human cells. The human D antigen represents a collection of epitopes expressed on a red cell membrane protein that is predicted to have 12 membrane‐spanning segments giving rise to six exofacial domains.STUDY DESIGN AND METHODS: By site‐directed mutagenesis using the method of inverse polymerase chain reaction, cE and D cDNA mutant constructs were generated with changes to the RHD‐specific residues 350, 353, and 354 in the predicted sixth exofacial loop. Each mutant cDNA was subcloned into the pBabe puromycin retroviral vector, and supernatants were used to transduce K562 cells. Puromycin‐resistant K562 clones were screened by flow cytometric analysis using a panel of monoclonal antibodies with specificities to ep (epitope) D1 through epD9.RESULTS: De novo expression of epD3 and epD9 was generated in the K562 cell lines expressing the mutated cE polypeptide (cE‐Asp350His, Gly353Trp, Ala354Asn). Expression of c and E was unaffected. Conversely, the cells expressing the mutated D polypeptide demonstrated loss of expression of epD1, epD2, epD3, epD4, and epD9.CONCLUSION: The data provide strong evidence for the critical involvement of three amino acids, Asp350, Gly353, and Ala354, in the expression of epD3 and epD9 on the predicted sixth external domain of the D protein. This domain also appears to be essential for the expression of epD1, epD2, and epD4, as a loss of expression of these epitopes was observed in K562 cells transduced with the Dmut construct (encoding His350, Trp353, and Asn354). The K562/Dmut, cell line has an identical molecular and serologic profile as the red cell DIVb pheno‐type, which confirms that retroviral gene transfer of Rh cDNA into K562 cells provides us with a powerful means by which to further map epitopes of D.
Measurement of circulating red cell volume using biotin‐labeled red cells: validation against51Cr‐labeled red cells
Authors: Mock Donald , Lankford Gary L. , Widness John A. , Burmeister Leon F. , Kahn Daniel , Strauss Ronald G. 
Published: March 2003

BACKGROUND:Anemia is a serious problem in the fetus and preterm infant. To investigate the physiology and pathophysiology of anemia and to assess responses to blood transfusions or erythropoietin therapy, measurement of circulating red cell volume would be useful. Because the standard51Cr method exposes the subject to radiation, a method of measuring circulating red cell volume without radiation exposure, sufficiently sensitive for use in fetuses and infants, was developed.STUDY DESIGN AND METHODS:In 10 healthy adults whose body mass ranged from 56.8 to 115.9 kg, aliquots of autologous red cells were labeled with biotin or with51Cr, mixed, and transfused intravenously. Circulating red cell volume was measured in posttransfusion blood by quantitating the in vivo dilution of biotinylated red cells. Biotinylated red cells were detected by two methods: 1)125I‐streptavidin and 2) fluorescein‐labeled avidin with flow cytometry.RESULTS:Circulating red cell volume measured by125I‐ streptavidin detection agreed well with that measured by51Cr (slope = 1.07, y‐intercept = −97, correlation = 0.987). Similarly, circulating red cell volume measured by flow cytometry agreed well with that measured by51Cr (slope = 1.05, y‐intercept = −20, correlation = 0.987).CONCLUSIONS:Circulating red cell volume measured by the use of biotin with either125I‐streptavidin or flow cytometry agrees with that measured by51Cr. Each system provides a method of performing these studies without exposing the subject to radiation.
Evaluation of the gel system for ABO grouping and D typing
Authors: Langston Melinda M. , Procter Jo Lynn , Cipolone Karen M. , Stroncek David F. 
Published: March 2003

BACKGROUND: The gel agglutination assay has been approved by the Food and Drug Administration as an alternative to the tube assay for the detection of red cell antibodies. It has also been approved recently by the Food and Drug Administration for ABO blood grouping and D typing.STUDY DESIGN AND METHODS: Tube and gel agglutination assays were compared for ABO grouping and D typing of 100 donor and 100 patient specimens. ABO grouping of 14 specimens of known ABO groups and D typing of 10 specimens with weak D were also compared. When antigen typing or isohemagglutinin results differed, gel testing was repeated by the use of modified incubation times, reagent or specimen volumes, and red cell concentrations.RESULTS: ABO grouping and D typing in all patient and donor specimens concurred. B isohemagglutinins were not detected in seven group A specimens. Six of seven discrepancies were resolved when gel tests were incubated at room temperature with increased serum or plasma volume. Weak D was detected in all 10 specimens tested by both assays. When weak A and/or B were tested with monoclonal antibody reagents, the correct phenotypes were identified in 9 specimens by gel assay and in 10 by tube assay. Using human antisera, 6 specimens were correctly phenotyped by gel assay and 7 by tube assay.CONCLUSION: The gel assay performed as well as the tube assay in detection of A, B, and D, but the tube assay was slightly better at detecting B isohemagglutinins. The gel assay can be used in place of the tube assay for ABO blood grouping and D typing.
Obtaining an accepted Investigational New Drug application to operate an umbilical cord blood bank
Authors: Reems Jo Anna , Fujita Deanna , Tyler Tom , Moldwin Richard , Smith Stephen 
Published: March 2003

BACKGROUND:The residual blood left in the placenta, previously considered a biologic waste, contains sufficient hematopoietic stem and progenitor cells to consistently engraft at least a small recipient. Over the past several years, more than 500 HLA‐matched, related and unrelated, allogeneic cord blood transplants have been performed. Consequently, public and private cord blood banks are being developed to meet future demands. Thus, the definition of a suitable and effective cord blood component needs to be critically defined. In February 1997, the US Food and Drug Administration (FDA) proposed that cord blood banks should operate under an Investigational New Drug (IND) license.STUDY DESIGN AND METHODS:Standard operating procedures were designed using standards from the Foundation for Accreditation of Hematopoietic and Cellular Therapy, the American Association of Blood Banks, and the National Marrow Donor Program and in accordance with current good manufacturing practices. The standard operating procedures were field‐tested and submitted to the FDA.RESULTS:Issues of the utmost concern to the FDA dealt with transplant recipient outcome data collection, donor recruitment, sample tracking, the use of unlicensed materials, and the reporting of positive infectious disease results. After three attempts, an IND application was approved.CONCLUSIONS:To obtain approval of an IND application, cord blood banks need a set of standard operating procedures that describe cord blood collection, processing, freezing, and storage. Issues relating to potential cord blood recipient identification, cord blood shipping, and reporting of transplant recipient outcomes are also needed. The IND process provides an opportunity for outside reviewers to make suggestions that may be included in the standard operating procedures.
A molecular approach for isolating high‐affinity Fab fragments that are useful in blood group serology
Authors: Czerwinski Marcin , Krop‐Watorek Anna , Siegel Don L. , Spitalnik Steven L. 
Published: March 2003

BACKGROUND: Multiple mouse hybridoma antibodies recognize the antigens of the MNS blood group system. The Fab fragments of several of these antibodies were expressed on bacteriophage and as soluble proteins. The parental N92 anti‐N IgG monoclonal antibody (parental N92 MoAb), but not its monovalent, soluble Fab fragment (N92 Fab fragment), agglutinated antigen‐positive red cells by an antiglobulin method. Light‐chain shuffling was used to isolate mutant N92 Fab fragments with higher affinity that would function by agglutination.STUDY DESIGN AND METHODS: Light‐chain cDNA libraries, constructed from mice immunized with N‐type glycophorin A, were inserted into a recombinant pComb3H vector containing the N92 Fd fragment. The N92 Fd fragment: light‐chain libraries were panned on N‐type glycophorin A or NN red cells, and antigen‐binding clones were isolated. Purified parental N92 MoAb and the Fab fragments were evaluated by enzyme‐linked immunosorbent assay and agglutination.RESULTS: The novel NNA7, C1, and G11 Fab fragments all bound to N‐type glycophorin A with higher affinity than did the N92 Fab fragment. The affinity of the library‐derived clones was equivalent to that of the parental N92 MoAb. Although their fine specificity differed slightly from the parental N92 MoAb, the clones functioned equivalently by agglutination using an antiglobulin method.CONCLUSIONS: Light‐chain shuffling allowed the isolation of bacterially produced, high‐affinity, soluble, monovalent recombinant anti‐N Fab fragments that functioned well by agglutination. This approach is useful in obtaining inexpensive serologic reagents that may replace conventional MoAbs produced by tissue culture methods.
Immune response to blood transfusion invery‐low‐birthweight infants
Authors: Wang‐Rodriguez J. , Fry E. , Fiebig E. , Lee T.‐H. , Busch M. , Mannino F. , Lane T.A. 
Published: March 2003

BACKGROUND: Allogeneic blood transfusion is common in the treatment of neonatal anemia of prematurity or anemia due to multiple phlebotomies. The immune response of neonates to passenger leukocytes from allogeneic red cells was investigated.STUDY DESIGN AND METHODS: Fourteen infants (4 male, 10 female) prospectively were randomly assigned to receive either white cell‐reduced (Group 1) or non‐white‐cell reduced (Group 2) irradiated blood. Blood samples were taken before and at various time intervals after transfusion (Days 1, 5‐7,and 10‐14). Cord blood from 11 healthy term infants was used for comparison. The following surface markers were used to assess immune modulation by flow cytometry: CD45RA/CD45RO, CD4/CD8, CD25/CD28, CD3/DR, CD14/B7, and CD3/CD56+CD16. Donor cell microchimerism was studied using semi‐quantitative polymerase chain reaction Y‐chromosome detection in female infants who received male donor blood. Donor and recipient HLA class II typing was performed with polymerase chain reaction with sequence specific primers.RESULTS: The lymphocyte counts in both groups were significantly increased after transfusion, and there was a significant increase in lymphocytes expressing CD45RA, CD3–/CD16+CD56, CD80, and CD3–/DR on Day 14. The premature infants' pretransfusion natural killer cell population (CD3–/CD16+CD56) was significantly lower than that of term infants, but it reached a similar level by Days 10‐14. CD8 subpopulations were increased but not CD4+ cells. Two female infants (of 6) had circulating Y chromosomes 1 day after transfusion, and most of the infants effectively cleared the donor cells within 24 hours of transfusion. Two Group 2 infants who by chance received presumably HLA‐haploidentical donor blood developed necrotizing enterocolitis.CONCLUSION: Blood transfusion alters immune cell antigen expression in premature neonates and may initially be immunostimulatory and later immunosuppressive.
Distinguishing full siblings from half‐siblings in limited pedigrees
Authors: Wenk R.E. , Chiafari F.A. 
Published: March 2003

BACKGROUND: Full siblings were compared with half‐siblings to observe how well the two relationships could be distinguished by informative tests.STUDY AND DESIGN METHODS: Parentage analysis ascertained 25 pairs of full siblings and 25 pairs of half‐siblings. The pairs were then examined for the sharing of alleles at three independent variable number of tandem repeat (VNTR) loci. A sibling index (SI) and a half‐sibling index (HSI) were calculated for each pair, and an SI:HSI ratio was determined.RESULTS: The SI:HSI ratio favored full siblings in 18 of 25 full sibling pairs. The SI:HSI ratio exceeded 100 in 8 of those 25 pairs. Although the ratio favored half‐siblings in 23 of 25 half‐sibling pairs, as was expected with the use of only 3 loci, it exceeded 0.1 in all 25 pairs.CONCLUSION: Study of more than three highly informative loci is required to improve the identification of full siblings and might well permit the identification of half‐siblings.
Filtration of RBC units:effect of storage time and temperature on filter performance
Authors: Smith J.D. , Leitman S.F. 
Published: March 2003

BACKGROUND: The influence of time, temperature, and rate of filtration on the efficacy of WBC reduction of RBC units was studied in a controlled, paired‐donor format.STUDY DESIGN AND METHODS: Ten donors underwent whole‐blood phlebotomy on two to four occasions each. Units were filtered (RCXL‐1, Pall Biomedical) under laboratory conditions and gravity flow as follows: 1) after 0 to 2 hours of storage at 22°C, 2) after 7 to 8 hours at 22°C, 3) after 14 days of storage at 4°C, and 4) under mock bedside conditions after 14 days of storage at 4°C. Prefiltration and postfiltration cell counts and prefiltration WBC CD11a expression were assessed on Days 0 and 14.RESULTS: WBC content before filtration was 2.20 and 2.34 x 109 (p>0.05) for units stored for 2 and 8 hours (Groups 1 and 2) and declined to 52.8 and 7.57 x 104 (p<0.01) after filtration. The efficacy of WBC reduction in units stored for 14 days was similar to that in units stored for 8 hours, but absolute postfiltration WBC counts were significantly lower because of a 0.6 log reduction in the starting WBC count after 14 days of storage (postfiltration WBC content of 1.02 and 2.31 x 104 for units filtered under laboratory vs. bedside conditions [p>0.05]). Filtration under bedside conditions was associated with a greater degree of variation in residual WBC counts than laboratory filtration. WBC reduction by filtration was significantly greater in units stored for at least 8 hours (Groups 2, 3, and 4) than in those stored for less than 2 hours (4.59 log vs. 3.83 log reduction in WBC content, p<0.05). Surface expression of leukocyte function antigen 1 as measured by CD11a was similar in all groups.CONCLUSION: WBC reduction of RBC units by filtration was least effective when performed within 2 hours of collection. Efficacy of WBC reduction increased significantly after the units were stored for 8 hours to 14 days, without significant differences between these storage intervals. Laboratory filtration yielded more consistent results than did mock bedside filtration. Temperature and filtration rate had no effect on the efficacy of WBC reduction by filtration.
Effects of the addition of second‐messenger effectors to platelet concentrates separated from whole‐blood donations and stored at 4°C or –80°C
Authors: Lozano M. , Escolar G. , Mazzara R. , Connor J. , White J.G. , DeLecea C. , Ordinas A. 
Published: March 2003

BACKGROUND: Platelet concentrates (PCs) are currently stored at 22°C under continuous agitation. Because of the potential risk of the overgrowth of bacteria in case of contamination, PC shelf life is limited to 5 days. A mixture of second‐messenger effectors is being evaluated to determine if it has benefits for cold liquid storage and cryopreservation of platelets.STUDY DESIGN AND METHODS: PCs separated from whole‐blood donations by the buffy coat method were randomly assigned (n = 6 each) to be stored for 5 days at 22°C under continuous agitation or at 4°C after treatment with a platelet storage medium (ThromboSol, LifeCell Corp.). PCs were also cryopreserved with 6‐percent DMSO (final concentration) or with ThromboSol plus 2‐percent DMSO (final concentration) (TC). After storage, platelets were analyzed by flow cytometry, transmission electron microscopy, and aggregation and perfusion techniques.RESULTS: Cold liquid storage of ThromboSol‐treated platelets resulted in a lower binding of coagulation factor Va on the platelet surface than on platelets stored at 22°C. In transmission electron microscopy, a conversion to spherical morphology was seen in the case of cold liquid storage. No difference between ThromboSol‐treated platelets stored at 4°C and platelets stored at 22°C was seen in perfusion studies. Cryopreservation in the presence of TC prevented the reduction in glycoprotein Ib and IV expression on platelet surface that is seen in 6‐percent DMSO‐cryopreserved platelets. Platelets cryopreserved in TC covered, by thrombus, a significantly greater percentage of the perfused surface after the freezing and thawing process.CONCLUSION: ThromboSol‐treated PCs separated from whole‐blood donations by the buffy coat method, stored at 4°C for 5 days, or cryopreserved in the presence of TC, maintained in vitro functional activity comparable to that achieved by current methods of storage, although discoid morphology was not preserved during cold liquid storage with ThromboSol.
Detection of multiple passively acquired alloantibodies following infusions of IV Rh immune globulin
Authors: Rushin J. , Rumsey D.H. , Ewing C.A. , Sandler S.G. 
Published: March 2003

BACKGROUND: Passively acquired blood group alloantibodies are detected regularly after infusions of IV Rh immune globulin (RhIG) for the treatment of immune thrombocytopenic purpura (ITP) in D+ patients.STUDY DESIGN AND METHODS: Blood samples from 16 D+ patients with ITP were tested after treatment with IV RhIG for the presence of passively acquired alloantibodies. Similar studies were conducted for three D– patients after injections of IM RhIG for Rh immunoprophyl‐axis. Four production lots of IV RhIG and 2 lots of IM RhIG were tested for the presence of alloantibodies.RESULTS: All 16 D+ patients with ITP developed a positive DAT, as well as positive antibody detection test results, after infusions of IV RhIG. All postinfusion plasma samples contained anti‐D, as well as one or more additional antibodies, usually anti‐C, ‐E, ‐G, ‐V, or ‐Fya. Eluates from patients' RBCs with positive DAT results contained multiple passively acquired alloantibodies. Multiple alloantibodies were detected in samples of different production lots of IV RhIG or IM RhIG. No acute transfusion reactions were observed in five D+ patients with ITP who had been treated with IV RhIG and had been given serologically incompatible D+ RBCs. After injections of IM RhIG, the only passively acquired alloantibody detected was anti‐D.CONCLUSION: Plasma samples from D+ patients with ITP treated with IV RhIG regularly contained anti‐D and multiple other passively acquired Rh, Duffy, or Kidd system alloantibodies. Postinfusion RBC samples all had positive DAT results with eluates containing anti‐D and multiple other Rh, Duffy, or Kidd system antibodies. The consistent detection of multiple passively acquired alloantibodies after IV RhIG, in contrast to the detection of anti‐D only after IM RhIG, reflects the immediate effect of the entire (bolus) dose of RhIG by the IV route, the dose for treating ITP that is approximately 10 times the dose for Rh immunoprophylaxis, and the expected serologic incompatibility with recipients' D+ RBCs.