bluuebunny/crossref_metadata_2025 huggingface dataset:
Aprotinin, cardiac surgery, and factor V Leiden
Authors: Sweeney J.D. , Blair A.J. , Dupuis M.P. , King T.C. , Moulton A.L.
Published: March 2003

BACKGROUND: Aprotinin has been shown to reduce blood transfusion in cardiac surgery. Aprotinin inhibits activated protein C (APC). Patients with factor V (FV) Leiden have an inherited resistance to APC proteolysis. If the inhibition of APC by aprotinin contributes to its beneficial effect in cardiac surgery, then patients with FV Leiden undergoing cardiac surgery might be expected to require less transfusion than patients without FV Leiden. However, the use of aprotinin in such patients also could compromise the protein C regulatory pathway and precipitate a clinical thrombotic event. STUDY DESIGN AND METHODS: Patients undergoing cardiac surgery were studied for the presence of the FV Leiden defect by the use of a Russell's viper venom clot‐based assay and polymerase chain reaction. The total amount of blood transfused was recorded for each patient. The effect of aprotinin on the plasma of normal and FV Leiden patients was studied. Further studies were performed on the direct inhibition of APC by aprotinin. RESULTS: Over an 18‐month period, 162 patients were studied, of whom 13 (8%; 95% CI, 4.3–13.3%) were positive for FV Leiden. These 13 had a smaller requirement for blood transfusion than the to 13 matched controls. In vitro, aprotinin induced a FV Leiden defect in normal plasma and exacerbated the defect in the plasma of FV Leiden patients. Aprotinin inhibited APC in a dose‐dependent manner, and kinetic analysis showed competitive inhibition with an inhibition constant of 4.5 microM (250 Kallikrein inhibitor units/mL). CONCLUSION: The inhibition of APC by aprotinin may contribute to its hemostatic effect. The use of aprotinin in patients with FV Leiden could cause extreme dysfunction of the protein C regulatory pathway, which could result in clinical thrombosis.
Transfusion transmission of human T‐lymphotropic virus type I (HTLV‐I) from an asymptomatic blood donor: conservation of LTR U3, env, and tax nucleotide sequences in a recipient with HTLV‐I‐associated myelopathy
Authors: Gasmi M. , D'Incan M. , Desgranges C.
Published: March 2003

BACKGROUND: Transfusion of human T‐lymphotropic virus type I (HTLV‐I) contaminated blood is sometimes linked with the rapid onset of HTLV‐I‐ associated myelopathy/tropical spastic paraparesis (HAM/TSP) in immunocompromised recipients.

STUDY DESIGN AND METHODS: This study addressed the question of whether HTLV‐I variants could emerge in an immunocompromised patient who developed HAM/ TSP after the transfusion of blood from an asymptomatic HTLV‐I carrier. Base pairs (n = 2327) of the HTLV‐I genome located in the LTR U3 region and parts of the env and tax genes were sequenced and compared to the isolated identified in the asymptomatic blood donor and to well‐known ATK‐1 and H5 strains. The same analysis was performed on another set of samples from an asymptomatic HTLV‐I‐positive blood donor and a similar blood recipient.

RESULTS: No critical changes in nucleotide sequences were identified in the immunosuppressed HAM/TSP patient when compared with the nucleotide sequences of the corresponding blood donor, the other asymptomatic blood donor and recipient or the ATK‐1 and H5 strains.

CONCLUSION: Immunosuppression does not seem to favor the emergence of particular nucleotide sequences in the genome located in the LTR U3 region or in parts of the env and tax genes in transfused patients who develop HAM/TSP.
Radiation‐induced red cell damage: role of reactive oxygen species
Authors: Anand A.J. , Dzik W.H. , Imam A. , Sadrzadeh S.M. H.
Published: March 2003

BACKGROUND: Cellular blood components are irradiated to prevent graft‐ versus‐host disease in transfusion recipients at risk for this syndrome. Because gamma radiation can result in the production of reactive oxygen species, the role of reactive oxygen species was investigated in radiation‐induced red cell damage. STUDY DESIGN AND METHODS: Whole blood from normal donors was exposed to various doses of t‐butyl hydroperoxide (0‐1 mM) and/or to gamma‐radiation (0‐50 Gy). Oxidative damage was assessed by the extent of lipid peroxidation (measured by thiobarbituric acid‐reactive substances [TBARS]) and hemoglobin oxidation. Fresh blood was divided into three parts‐one initially irradiated and stored, another stored with portions irradiated weekly, and a third stored without irradiation. TBARS and hemoglobin oxidation were measured weekly. RESULTS: As expected, t‐ butyl hydroperoxide induced TBARS formation and hemoglobin oxidation in a dose‐dependent fashion. The gamma‐radiation not only increased hemoglobin oxidation and TBARS formation, but also enhanced the t‐butyl hydroperoxide effect on red cells. Red cell storage increased TBARS generation and hemoglobin oxidation in a time‐dependent fashion. When radiation was administered either initially or after weekly storage, TBARS production and hemoglobin oxidation were increased over that measured in unirradiated paired controls. CONCLUSION: Gamma radiation at clinically used doses increases lipid peroxidation and hemoglobin oxidation in human red cells. The effect of gamma‐radiation is accentuated by blood storage and induces damage independent of time of storage.
Activation of the contact system by filtration of platelet concentrates with a negatively charged white cell‐removal filter and measurement of venous blood bradykinin level in patients who received filtered platelets
Authors: Shiba M. , Tadokoro K. , Sawanobori M. , Nakajima K. , Suzuki K. , Juji T.
Published: March 2003

BACKGROUND: Several recent reports have described hypotensive transfusion reactions in patients receiving platelet concentrates (PCs) filtered through white cell‐reduction filters. It is well known that a negatively charged surface activates the contact system, consisting of factor XII, prekallikrein, and high‐molecular‐weight kininogen.

STUDY DESIGN AND METHODS: To clarify the mechanisms of these hypotensive reactions, the possibility that white cell‐reduction filtration activates the contact system was examined. Venous blood plasma bradykinin levels were also measured in patients receiving PC transfusions through filters.

RESULTS: None of the measured values were changed by filtration through a positively charged filter. However, filtration through a negatively charged filter resulted in a decrease in the amounts of prekallikrein and an increase in the amount of bradykinin generated, which indicated the activation of the contact system. The bradykinin level was inversely related to the activity of angiotensin‐converting enzyme (ACE) in the PCs and was elevated by addition of an ACE inhibitor. Although the venous blood plasma bradykinin level did not change in two patients with a normal ACE activity during PC transfusion through the negatively charged filter, two patients who had decreased ACE activity, showed a significant increase in bradykinin during the transfusion.

CONCLUSION: These results suggest that the generation of a large amount of bradykinin by filtration of PCs through a negatively charged filter might cause hypotensive reactions in patients with decreased ACE activity. The clinical significance of bradykinin generation requires further study.
Autologous donation error rates in Canada
Authors: Goldman M. , Rémy‐Prince S. , Trépanier A. , Décary F.
Published: March 2003

BACKGROUND: Although certain transfusion risks are eliminated by the use of autologous blood, clerical errors may still occur. In addition, because of differences in donor selection criteria and donor‐patient expectations, the consequences of certain errors may be different in autologous and allogeneic donations.

STUDY DESIGN AND METHODS: In January 1996, autologous donation error rates in Canada from 1989 to November 1995 were estimated by 1) a detailed questionnaire sent to hospitals supplied by the Canadian Red Cross, Blood Services, Transfusion Center of Quebec at Montreal autologous donation program (n = 31), 2) a review of that institution's quality assurance non‐ compliance reports, and 3) a detailed questionnaire sent to other Canadian Red Cross centers with autologous donation programs (n = 16) and hospital‐based autologous programs in Canada (n = 3). The total number of autologous donations collected was determined from Canadian Red Cross annual reports and information supplied by hospital‐based programs.

RESULTS: There were 113 errors reported for 16,873 units collected by the Montreal center (1/149 units) based on collection center and hospital data. The most frequent errors were the late receipt of units for surgery (25% of errors) or the receipt of units in the wrong hospital (23%). Other Canadian programs reported 166 errors for approximately 53,500 units collected (1/322 units). However, this figure was based mainly on collection center, and not hospital, data. The most frequent errors were in labeling (48%) and component preparation (25%). One unit of autologous fresh‐frozen plasma was transfused to the wrong recipient. Errors were more frequent if components were produced, if units were drawn in hospitals for interhospital transfer, or it units were shipped between Red Cross centers.

CONCLUSION: Errors are not infrequent in autologous donation programs. Autologous transfusion should not be considered as being without risk.
The association of biologically active lipids with the development of transfusion‐related acute lung injury: a retrospective study
Authors: Silliman Christopher C. , Paterson Andrew J. , Dickey William O. , Stroncek David F. , Popovsky Mark A. , Caldwell Sherrie A. , Ambruso Daniel R.
Published: March 2003

BACKGROUND: Transfusion‐related acute lung injury (TRALI) is clinically similar to the adult respiratory distress syndrome (ARDS) and has been linked to the transfusion of leukocyte antibodies in blood components. Animal model have implicated neutrophil (PMN)‐priming agents in ARDS; however, two agents were required. Previous studies showed the generation of PMN‐priming agents during blood storage. Thus the association of PMN‐priming agents with TRALI was examined. STUDY DESIGN AND METHODS: Ten patients with TRALI and 10 with febrile or urticarial reactions (control group) were evaluated. The presence of PMN‐priming activity was tested in the patients' pretransfusion and posttransfusion blood samples by incubating PMNs with these samples followed by activation of the respiratory burst. Plasma lipids were separated by normal‐phase high‐performance liquid chromatography (HPLC), and the priming activity was evaluated. The presence of leukocyte antibodies was determined in the blood donors and patients with TRALI. RESULTS: Significantly more PMN‐priming activity was present in the posttransfusion sera (11.4 +/− 1.8 nmol superoxide anion/min, mean +/− SEM; n=10) and plasma of patients with TRALI than in their pretransfusion sera (6.5 +/− 1.5: n=10) or in the pretransfusion and posttransfusion sera (5.1 +/− 1.3, n=10; and 4.5 +/− 1.4, n=10, respectively) and from the controls (p < 0.05). HPLC separation of lipids demonstrated that three active species were present in the posttransfusion plasma samples of TRALI patients. All the patients with TRALI had underlying clinical factors, such as infection, cytokine administration, recent surgery, or massive transfusion, while only 2 of 10 control patients had these clinical conditions. None of the donors had significant titers of HLA or HLA‐DR antibodies; however, 50 percent had weak positivity for granulocyte antibodies. CONCLUSION: TRALI is the result of two clinical events, the first being a predisposing clinical condition and the second being the transfusion of biologically active lipids in stored blood.
Hemostasis in patients with severe von Willebrand disease improves after normal platelet transfusion and normalizes with further correction of the plasma defect
Authors: Castillo Ricardo , Escolar Ginés , Monteagudo Juan , Aznar‐Salatti José , Reverter Juan Carlos , Ordinas Antonio
Published: March 2003

BACKGROUND: A defective hemostatic effect of plasma concentrate infusion in patients with severe von Willebrand disease (vWD) has been ascribed to the absence of platelet von Willebrand factor (vWF) STUDY DESIGN AND METHODS: The role of platelet vWF in hemostasis of severe vWD was investigated. A plateletpheresis unit (4‐5 × 10(11) platelets) from a normal compatible donor was transfused before any cryoprecipitate infusion to three type 3 vWD patients and to one patient with severe type 1 vWD with low levels of platelet vWF who required replacement therapy for bleeding episodes. Autologous platelets were transfused to one of the patients with type 3 vWD. RESULTS: Partial corrections of bleeding times (14‐17 min vs. baseline>30 min) were observed in all patients after the transfusion of normal platelets. During cryoprecipitate infusion, bleeding times were normalized (<6 min), and bleeding episodes stopped when plasma levels of vWF activity ranged from 14 to 18 U per dL. Platelet interactions with the subendothelium increased in parallel with the correction of bleeding times. These results indicate that if approximately 20 percent of the total number of platelets have normal vWF antigen and if plasma vWF levels are at least 14 U per dL, then bleeding times will normalize and mucosal hemorrhages will stop. Transfusion of autologous platelets in one patient with type 3 vWD did not modify bleeding times or platelet adhesion on the subendothelium. CONCLUSION: The hemostatic effect of normal platelets in type 3 vWD seems to be related to the platelet vWF in the transfused platelets.
Report on the second international granulocyte serology workshop
Authors: Bux J. , Chapman J.
Published: March 2003

BACKGROUND: The methods of granulocyte antibody and antigen detection have improved considerably since the First International Granulocyte Serology Workshop. STUDY DESIGN AND METHODS: Thirteen laboratories participated in the Second International Granulocyte Serology Workshop. The study was designed to meet five goals: 1) establishment of antigen‐ typed granulocyte panels, 2) determination of the proficiency of granulocyte antibody detection by laboratory and by technique, 3) identification of granulocyte antibodies present in uncharacterized sera, 4) genotyping of NA antigens by DNA‐based techniques, and 5) collection and exchange of information to standardize granulocyte antibody screening. RESULTS: NA1‐, NA2‐ and NB1‐specific antibodies were detected by more than two‐thirds of the participants with the granulocyte immunofluorescence test (GIFT) but by fewer than two‐thirds with the granulocyte agglutination test (GAT). The determination of the NB2‐, 5b‐, and Fc gamma RIIIb‐specific antibodies was the most problematic. Ninety percent of the participants were not able to identify NB2 antibodies in sera with suspected NB2 specificity. The 5b antibodies were only detected by laboratories that performed the GAT. Isoantibodies to Fc gamma RIIIb were identified when the monoclonal antibody‐specific immobilization of granulocyte antigens (MAIGA) assay or an Fc gamma RIIIb‐deficient panel cell was available. In 4 of 11 uncharacterized sera, the presence of NA1, NA2, and Fc gamma RIIIb antibodies could be confirmed by the MAIGA assay. In polymerase chain reaction with sequence‐specific primers (PCR‐SSP), the NA genotypes of all DNA samples were correctly determined. CONCLUSION: A combination of the GIFT and GAT is still the best means of antibody detection. The MAIGA assay allows identification of isoantibodies to Fc gamma RIIIb or alloantibodies to NA antigens. The NA genotype can be reliably determined by PCR‐SSP. Cell panels should cover NA1, NA2, NB1, 5b, and, if possible, SH.
The VS and V blood group polymorphisms in Africans: a serologic and molecular analysis
Authors: Daniels G. L. , Faas B.H.W. , Green C. A. , Smart E. , Maaskant‐van Wijk P. A. , Avent N. D. , Zondervan H. A. , von dem Borne A. E. , van der Schoot C. E.
Published: March 2003

BACKGROUND: VS and V are common red cell antigens in persons of African origin. The molecular background of these Rh system antigens is poorly understood.

STUDY DESIGN AND METHODS: Red cells from 100 black South Africans and 43 black persons from Amsterdam, the Netherlands, were typed serologically for various Rh system antigens. Allele‐specific polymerase chain reaction and sequencing of polymerase chain reaction products were used to analyze C733G (Leu245Val) and G1006T (Gly336Cys) polymorphisms in exons 5 and 7 of RHCE and the presence of a D‐CE hybrid exon 3.

RESULTS: The respective frequencies of all VS+ and of VS+ V‐(r's) phenotypes were 43 percent and 9 percent in the South Africans and 49 percent and 12 percent in the Dutch donors. All VS+ donors had G733 (Val245), but six with G733 were VS‐ (4 V+w, 2 V‐). The four VS‐ V+w donors with G733 appeared to have a CE‐D hybrid exon 5. T1006 (Cys336) was present in 12 percent and 16 percent of donors from the two populations. With only a few exceptions, T1006, a D‐CE hybrid exon 3, and a C410T (Ala137Val) substitution were associated with a VS+ V‐phenotype ((C)ces or r's haplotype). Two VS+ V‐individuals, with the probable genotype, (C)ces/(C)ces), were homozygous for G733 and for T1006.

CONCLUSIONS: It is likely that anti‐VS and anti‐V recognize the conformational changes created by Val245, but that anti‐V is sensitive to additional conformational changes created by Cys336.
The distribution of infectivity in blood components and plasma derivatives in experimental models of transmissible spongiform encephalopathy
Authors: Brown P. , Rohwer R. G. , Dunstan B. C. , MacAuley C. , Gajdusek D. C. , Drohan W. N.
Published: March 2003

BACKGROUND: The administration of blood components from donors who subsequently develop Creutzfeldt‐Jakob disease has raised the issue of blood as a possible vehicle for iatrogenic disease.

STUDY DESIGN AND METHODS: We examined infectivity in blood components and Cohn plasma fractions in normal human blood that had been “spiked” with trypsinized cells from a scrapie‐infected hamster brain, and in blood of clinically ill mice that had been inoculated with a mouse‐adapted strain of human transmissible spongiform encephalopathy. Infectivity was assayed by intracerebral inoculation of the blood specimens into healthy animals.

RESULTS: Most of the infectivity in spiked human blood was associated with cellular blood components; the smaller amount present in plasma, when fractionated, was found mainly in cryoprecipitate (the source of factor VIII) and fraction I+II+III (the source of fibrinogen and immunoglobulin); almost none was recovered in fraction IV (the source of vitamin‐K‐dependent proteins) and fraction V (the source of albumin). Mice infected with the human strain of spongiform encephalopathy had very low levels of endogenous infectivity in buffy coat, plasma, cryoprecipitate, and fraction I+II+III, and no detectable infectivity in fractions IV or V.

CONCLUSION: Convergent results from exogenous spiking and endogenous infectivity experiments, in which decreasing levels of infectivity occurred in cellular blood components, plasma, and plasma fractions, suggest a potential but minimal risk of acquiring Creutzfeldt‐Jakob disease from the administration of human plasma protein concentrates.
Effect of 24‐hour whole‐blood storageon plasma clotting factors
Authors: O'Neill E.M. , Rowley J. , Hansson‐Wicher M. , McCarter S. , Ragno G. , Valeri C.R.
Published: March 2003

BACKGROUND: The current requirements for the preparation of fresh‐frozen plasma within 8 hours of whole‐blood collection were designed to maintain clotting factor activities. These requirements, however, limit the production of fresh‐frozen plasma in a large blood center. There are few data on the effect of the extension of CPD whole‐blood storage to 24 hours on clotting factor activity.

STUDY DESIGN AND METHODS: A 500‐mL unit of whole blood was collected from 10 volunteer donors. At 1 hour after collection, a plasma sample was separated by centrifugation, and each unit was equally divided into 2 half‐units, with 1 half‐unit stored at 4°C (range, 1‐6°C) and 1 half‐unit stored at 22 °C (range, 20‐24°C) for 8 hours after collection. Each half‐unit was then placed at 4°C for further storage for 16 hours. At 8 and 24 hours after collection, plasma samples were separated from each half‐unit. All plasma samples were frozen at –18°C. Factors V, VII, VIII, and X; fibrinogen; antithrombin III; protein C; and protein S were measured.

RESULTS: No significant changes were noted in factors V, VII, and X; fibrinogen; antithrombin III; protein C; and protein S over the 24‐hour storage period. Factor VIII in both half‐units was significantly reduced, by 13 percent, from the baseline sample as compared to the level in the 8‐hour storage sample (p<0.05). Factor VIII was further reduced by 15 to 20 percent after the 24‐hour storage period (p<0.05).

CONCLUSION: The coagulation factor activity for all factors measured, with the exception of factor VIII, showed no significant change over the 24‐hour storage period. Factor VIII was significantly decreased by 13 percent in 8‐hour storage and by an additional 15 to 20 percent in 24‐hour storage. For clinical situations not requiring the replacement of factor VIII only, 24‐hour frozen plasma has properties comparable to those of fresh‐frozen plasma.
Redefining the HIV‐infectious window period in the chimpanzee model: evidence to suggest that viral nucleic acid testing can prevent blood‐borne transmission
Authors: Murthy K.K. , Henrard D.R. , Eichberg J.W. , Cobb K.E. , Busch M.P. , Allain J.P. , Alter H.J.
Published: March 2003

BACKGROUND: Although it is rare, blood‐transmitted HIV infection can occur when a donor presents in the window period between HIV‐1 exposure and the first appearance of detectable p24 antigen.

STUDY DESIGN AND METHODS: To study this seronegative window period, a chimpanzee (X034) was inoculated with 38 median tissue culture infective doses of HIV‐1 IIIB; serum and peripheral blood mononuclear cells were obtained one to two times per week for 12 weeks and then biweekly for 12 weeks. Infectivity was monitored by the detection of serum HIV RNA, cell‐associated HIV DNA, p24 antigen, and anti‐HIV and by co‐culture methods.

RESULTS: No HIV markers were noted until 5 weeks after inoculation, at which time virus was isolated and HIV RNA and DNA were detected in plasma and cells, respectively. Anti‐HIV and HIV p24 antigen were not present until 8 weeks after inoculation. Plasma and cells obtained from Chimpanzee X034 3 or 4 weeks after exposure were then sequentially inoculated into a second chimpanzee (X176); no HIV infection was observed in this animal during serial follow‐up for 24 weeks after each inoculation. In contrast, when the fifth‐week HIV‐1 RNA‐ and DNA‐positive sample was inoculated, Chimpanzee X176 was unequivocally infected with HIV‐1.

CONCLUSIONS: Nucleic acid testing narrowed the seronegative window by 3 weeks (37%). More important, there was no demonstrable infectivity in either plasma or peripheral blood mononuclear cells obtained before molecular markers were detectable. This suggests that the infectious window may be considerably shorter than the total window as measured from exposure and that nucleic acid testing might not only shorten the seronegative window, but totally prevent transfusion‐transmitted HIV infection.
Determination of reasons for cessation of participation in serial plasmapheresis programs
Authors: Rodell M.B. , Lee M.L.
Published: March 2003

BACKGROUND: An earlier retrospective review of records of over 500 remunerated donors participating in plasmapheresis programs at various levels of frequency for 10 or more years showed that there were no significant differences in mean values for total protein, microhematocrit, body weight, and individual serum proteins, regardless of the number of annualized donations for each of four frequency groups. Although these data demonstrated that donors undergoing serial plasmapheresis for 10 or more years were able to successfully continue in such programs, the review did not include information relative to individuals who had discontinued their participation. The current study investigated reasons that donors drop out of plasmapheresis programs.

STUDY DESIGN AND METHODS: Donors who had successfully undergone plasmapheresis at least 20 times within a recent 6‐month period but who had failed to appear for at least 30 days subsequent to their last donation were defined as potential subjects for the study. Staff members at the participating center contacted the donors, inviting them to return to the center to participate in a survey. Donors were also asked for a blood sample upon their arrival at the plasma center; samples were tested for total and various serum proteins.

RESULTS: A total of 409 eligible subjects appeared at the centers for the survey, and 375 blood samples were analyzed for proteins. Survey responses indicated that the predominant reasons for an end to participation were socioeconomic in nature. Serum protein findings were compared to the most recent results obtained while the donors were active in the plasmapheresis program. Slightly higher mean serum protein findings were seen for samples collected on the survey date.

CONCLUSION: A review of the aggregate responses obtained in the survey indicates that the predominant reasons that donors ceased to participate were not health‐related: the remuneration was no longer needed, and work schedule conflicts, time constraints, and relocation were frequently cited.
The dose of granulocyte–colony‐stimulating factor after chemopriming treatment does not influence apheresis yield of progenitor cells: a retrospective study of 91 cases
Authors: Lefrère F. , Belanger C. , Audat F. , Hermine O. , Cavazzana‐Calvo M. , Arnulf B. , Buzyn A. , Varet B.
Published: March 2003

BACKGROUND: The optimal dose of post‐chemotherapy granulocyte–colony‐stimulating factor (G–CSF) administration before peripheral blood progenitor cell (PBPC) collection has not been determined as yet, although 5 μg per kg per day has been recommended as the standard dose. This study retrospectively analyzed the effect of G–CSF dose on peripheral blood CD34+ cell collection from 91 patients with hematologic malignancies.

STUDY DESIGN AND METHODS: Various doses of G–CSF were administered after several chemotherapeutic PBPC mobilization regimens. According to the dose of G–CSF administered, patients were assigned to two groups. Group 1 included 46 patients who received a low dose of G–CSF (median, 3.6 [range, 2.8‐4.6] μg/kg/day). Group 2 included 45 patients who received a standard G–CSF dose of 6.0 (5.5‐8.1) μg per kg per day. Patients in the two groups were matched for age, diagnosis, previous therapy, and chemotherapeutic PBPC mobilization regimens.

RESULTS: No difference was observed in the median number of CD34+ cells harvested from each group. The number of leukapheresis procedures necessary to obtain a minimum of 3 × 106 CD34+ cells per kg was the same in both groups, and the percentage of patients who failed to achieve adequate PBPC collections was similar in the two groups.

CONCLUSION: The administration of low‐dose G–CSF after chemotherapy appears equivalent to administration of the standard dose in achieving satisfactory PBPC collection. This approach could allow significant savings in medical cost. A randomized and prospective study is necessary, however, to assess the validity of these conclusions.
Clinical evaluation of a solid‐phase test for red cell antibody screening of pregnant women
Authors: Shanwell Agneta , Sallander Suzanne , Bremme Katarina , Westgren Magnus
Published: March 2003

BACKGROUND: The aims of this study were to evaluate the results of a new solid‐phase screening test for detecting atypical red cell (RBC) antibodies in a large number of pregnant women and to compare these results to the clinical outcome of the newborn.

STUDY DESIGN AND METHODS: A total of 38,700 infants born in Stockholm were studied retrospectively. Of these infants, 18,500 were born to pregnant women screened with the solid‐phase test. Data were collected on all newborns with a positive direct antiglobulin test (DAT) and on infants requiring an exchange transfusion or a blood transfusion. These data were correlated to the screening results for the mothers.

RESULTS: Of 409 DAT‐positive newborns, a serologic explanation for the positive DAT was found in 349. Three hundred four cases were due to ABO incompatibility between mother and child; 19 of these infants needed an exchange transfusion. Forty‐two cases were due to unexpected maternal RBC antibodies; 11 of these infants were given an exchange transfusion. All 11 were identified before birth. Three other infants had DAT‐positive tests due to ABO incompatibility and to unexpected maternal RBC antibodies.

CONCLUSION: ABO incompatibility is a major indication for exchange transfusion in DAT‐positive newborns. There was no evidence that the solid‐phase screening test had failed to detect any clinically significant RBC antibodies. Finally, the results of this study do not indicate a need for routine screening of D+ women more than once during each pregnancy.
Prevalence of HLA sensitization in female apheresis donors
Authors: Densmore Tamara L. , Tim Goodnough Lawrence , Ali Suhail , Dynis Marian , Chaplin Hugh
Published: March 2003

BACKGROUND: Transfusion‐related acute lung injury (TRALI) is a serious complication of plasma‐containing blood components. Studies have implicated HLA antibodies along with biologically active lipids in stored blood in the pathogenesis of TRALI. It has been proposed that the exclusion of HLA‐untested, multiparous donors of plasma‐rich components, including plasma and single‐donor apheresis platelets, would substantially reduce the risk of TRALI.

STUDY DESIGN AND METHODS: To investigate the feasibility of such an exclusion, 332 female plateletpheresis donors with a record of over 9000 donations, none of which were associated with TRALI, were studied.

RESULTS: Seventeen percent of female donors demonstrated HLA sensitization. Parity and HLA sensitization were significantly correlated (p<0.0001), with sensitized donors having an average of 2.9 (± 0.6 95% CI) prior pregnancies and unsensitized donors having an average of 1.8 (± 0.2 95% CI) prior pregnancies. The percentage of HLA‐sensitized women with 0, 1 to 2, and >3 pregnancies was 7.8, 14.6, and 26.3, respectively.

CONCLUSION: These findings confirm the hypothesis that multiparous women (>3 pregnancies) represent an increased potential risk for TRALI. However, the exclusion of multiparous plateletpheresis donors would eliminate one‐third of our female donor pool. Screening such donors for HLA sensitization may represent the optimal approach for identifying donors at risk for causing TRALI, but this still would result in the deferral of 8 percent of female plateletpheresis donors. At present, prospective screening to identify donors at risk for causing TRALI is not justified.
Greater increase in cytokine concentration after salvage with filtered whole blood than with washed red cells, but no difference in postoperative hemoglobin recovery
Authors: Åvail Anders R. , Hyllner Monica , Bengtson Jan Peter , Carlsson Lars , Bengtsson Anders
Published: March 2003

BACKGROUND: Inflammatory mediators are released in association with intraoperative and postoperative salvage of blood. Whether these mediators (cytokines) participate in the modulation of erythropoiesis or not has been investigated.

STUDY DESIGN AND METHODS: Twenty‐seven patients who were to undergo total knee replacement surgery were randomly assigned to postoperative blood salvage with either filtered whole blood or washed red cells. Patients with postoperative blood loss <400 mL were considered a control group. The control group did not receive any transfusions. Plasma concentrations of the anaphylatoxin C3a, the C5b‐9 terminal complement complex, and the cytokines interleukins 6 and 8, hemoglobin, reticulocytes, and red cell volume fraction in the patients were repeatedly analyzed before and after surgery.

RESULTS: Significantly increased concentrations of interleukin 6 appeared in all three groups, which was interpreted as a response to the surgical trauma. The increase was significantly greater in the group that received filtered whole blood after return of shed blood. The recovery of hemoglobin levels did not differ in the groups.

CONCLUSION: The transfusion of filtered whole blood leads to the formation of interleukin 6 in the circulation, but postoperative hemoglobin recovery was similar in all groups.
Hydroxyethyl starch reduces the chemotaxis of white cells through endothelial cell monolayers
Authors: Hofbauer Roland , Moser Doris , Hornykewycz Stephan , Frass Michael , Kapiotis Stylianos
Published: March 2003

BACKGROUND: Polymorphonuclear leukocytes (PMNs) play a tremendous role during inflammatory processes. PMNs have to pass a monolayer of endothelial cells to migrate into the extravascular space. Hydroxyethyl starch (HES) is frequently used as a volume expander in critically ill patients.

STUDY DESIGN AND METHODS: The aim of this study was to investigate whether HES influences the chemotaxis of PMNs through endothelial cell monolayers by using a test system that allows the simultaneous treatment of both cell types. Human umbilical endothelial cells were cultured on microporous membrane filters. PMNs were isolated and PMN chemotaxis was studied.

RESULTS: The number of untreated PMNs that migrated through untreated endothelial cell monolayers in response to a chemoattractant was used as a control and set as 100 percent. In clinically relevant concentrations, HES was able to significantly decrease PMN chemotaxis through endothelial cell monolayers, showing a dose‐dependent effect (0.1 mg/mL: 99.6 ± 10.9%, p = NS compared to control; 1 mg/mL: 82.4 ± 8.3%, p<0.05 compared to control; 10 mg/mL: 62.9 ± 11.7%, p<0.05).ln this assay, both cell types (PMNs and endothelial cells in the monolayer) were treated simultaneously, which simulated the clinical situation after an intravenous injection of HES. The treatment of one cell type, PMNs (89.6 ± 8.8%, p<0.05) or endothelial cells in the monolayer (76.2 ± 9.4%, p<0.05), suggests that the influence on endothelial cells is greater.

CONCLUSION: HES is able to significantly reduce the chemotaxis of PMNs through endothelial cell monolayers. The possible clinical consequence of a moderate reduction in endothelium‐mediated PMN chemotaxis in critically ill patients remains to be evaluated.
The number of platelet glycoprotein Ia molecules is associated with the genetically linked 807 C/T and HPA‐5 polymorphisms
Authors: Corral Javier , Rivera José , González‐Conejero Rocio , Vicente Vicente
Published: March 2003

BACKGROUND: The neutral 807 C/T (Phe224) polymorphism (807 C/T polymorphism) of the glycoprotein (GP)Ia gene has been recently associated with the number of GPIa molecules on the platelet surface. The association of the number of GPIa molecules with other GPIa polymorphisms, such as HPA‐5 (Glu/Lys505) (HPA‐5 polymorphism), involved in alloimmune thrombocytopenias is unknown.

STUDY DESIGN AND METHODS: The association of the HPA‐5 polymorphism with the number of GPIa molecules on the platelet surface in 159 white blood donors was investigated. The genetic linkage between the HPA‐5 and the 807 C/T polymorphisms in 316 individuals was also determined.

RESULSTS: Both the 807 C/T and HPA‐5 polymorphisms correlate with the number of GPIa molecules on the platelet surface. The 807 T and HPA‐5b alleles are associated with increased numbers of GPIa molecules on the platelet surface. Moreover, the HPA‐5b allele is genetically linked to 15.8 percent of the 807 C alleles. Therefore, the number of GPIa molecules on the platelet surface is dependent on both GPIa polymorphisms as follows: 807 T/T, HPA‐5 a/a > 807 C/T, HPA‐5 a/b > 807 C/T, HPA‐5 a/a > 807 C/C, HPA‐5 a/b > 807 C/C, HPA‐5 a/a.

CONCLUSION: Two GPIa polymorphisms (807 C/T and HPA‐5) responsible for the variability in the numbers of GPIa/lla molecules on the platelet surface in whites have been identified. Despite the genetic linkage between the two polymorphisms, their influence on the number of GPIa molecules on the platelet surface may occur through different mechanisms.
TT virus infections among blood donors in Iceland:prevalence, genotypes, and lack of relationship to serum ALT levels
Authors: Löve A. , Stanzeit B. , Li L. , Ólafsdóttir E. , Gudmundsson S. , Briem H. , Widell A.
Published: March 2003

BACKGROUND: The TT virus (TTV) is a newly identified blood‐borne virus. Its association with disease is still unknown, and screening of blood donors has not been implemented. Several genotypes of the TTV have been identified.

STUDY DESIGN AND METHODS: Three hundred seventy healthy blood donors were randomly selected and tested for TTV by the PCR method. Sequencing of a part of the genome was performed to identify various genotypes of the virus. ALT levels were determined in both infected and uninfected individuals.

RESULTS: The TT virus (TTV), was detected in the sera of 23 (6.2%) of 370 healthy Icelandic blood donors; this prevalence is lower than that reported in Japan but higher than that in Scotland. The virus was found in all groups over the age of 19. Sequencing and phylogenetic analysis of 202 bp from open reading frame 1 demonstrated genotypes 1b and 2b 2c and genotype 4 isolates, with the latter bearing 89‐percent nucleotide homology with other genotype 4 sequences deposited at GenBank. One sample showed a mixed genotype 1b/2c infection. Serum ALT levels were within normal limits in all infected individuals.

CONCLUSION: The TTV carrier state does not cause significant liver injury.
A multicenter study evaluating three methods for counting residual WBCs in WBC‐reduced blood components: Nageotte hemocytometry, flow cytometry, and microfluorometry
Authors: Dzik S. , Moroff G. , Dumont L. ,   for the Biomedical Excellence for Safer Transfusion (BEST) Working Party of the ISBT
Published: March 2003

BACKGROUND: A multicenter study was conducted to evaluate the performance characteristics of flow cytometry and microfluorimetry for counting low concentrations of WBCs and to compare the results with Nageotte hemocytometry.

STUDY DESIGN AND METHODS: A two‐phase study involving 10 centers located in the United States and in Europe was performed. Coded samples of RBCs and platelets were distributed by 24‐hour (Phase 1) or 2‐day (Phase 2) courier service to each test site for analysis. Samples were prepared to include concentrations of WBCs slightly above and below the concentration corresponding to the threshold standards for WBC‐reduced RBCs and platelets. All centers tested samples by Nageotte hemocytometry plus one or both of two automated methods.

RESULTS: Both flow cytometry and microfluorometry gave better results than Nageotte hemocytometry in testing freshly prepared samples. At WBC concentrations >5 per μL (RBCs) or >3 per μL (platelets), the intersite CV was <20 percent for the automated methods but >30 percent for the Nageotte hemocytometer method (p<0.001). Accuracy was greater for the automated methods than for the Nageotte hemocytometer method (p<0.001). Nageotte hemocytometry showed a bias to underestimation relative to the results obtained with the automated methods. All methods had poorer performance in testing samples that required ≥2 days' shipment than in testing of those requiring overnight shipment.

CONCLUSION: Automated methods for counting residual donor WBCs in WBC‐reduced cellular components offer advantages of improved precision and greater accuracy than are seen with the Nageotte hemocytometer method. Automated methods are less labor‐intensive but more costly than microscopic methods. Preparation and shipping methods will need further refinement for samples to be counted more than 24 hours after sample collection.
Immune‐mediated agglutination of cytoskeleton‐free RBC microvesicles
Authors: Jaime J.C.
Published: March 2003

BACKGROUND: The factors contributing to RBC agglutination are complex. The RBC cytoskeleton's participation in and contribution to this phenomenon are difficult to separate from those of the plasma membrane. Immunoreactive, cytoskeleton‐free, band 3‐enriched microvesicles can be generated from normal RBCs. Band 3 has been defined as an important antigen in autoimmune hemolytic anemia (AIHA).

STUDY DESIGN AND METHODS: RBC microvesicles devoid of major cytoskeletal proteins were generated and sensitized with eluates obtained from AIHA patients, DAT‐positive blood donors, and antisera to common RBC antigens. Monoclonal anti‐human IgG was added and agglutination was investigated. Autoantibody‐specific binding was evaluated by employing 125I protein A.

RESULTS: RBC vesicle agglutination with a 4+ anti‐human globulin score was obtained with 10 autoantibody eluates from AIHA patients and anti‐D (3+), but not with eluates from 20 DAT‐positive blood donors or antisera directed to eight other common RBC antigens. Microvesicles sensitized with AIHA eluates bound 67 to 167 times as much 125I protein A radioactivity as did those incubated with buffered normal saline and 18 to 45 times more than vesicles incubated with normal serum.

CONCLUSION: The major proteins of the RBC cytoskeleton are not required for supporting IgG immune‐mediated agglutination of RBC microvesicles.
Passenger lymphocyte syndrome with severe hemolytic anemia due to an anti‐Jka after allogeneic PBPC transplantation
Authors: Leo A. , Mytilineos J. , Voso M.T. , Weber‐Nordt R. , Liebisch P. , Lensing C. , Schraven B.
Published: March 2003

BACKGROUND: After allogeneic peripheral blood progenitor cell (PBPC) transplantation, a patient developed a severe hemolytic transfusion reaction due to passenger lymphocyte syndrome.

CASE REPORT: A 50–year‐old woman with secondary acute myeloid leukemia transforming from a myelodysplastic syndrome received an ABO‐compatible PBPC graft from her HLA‐identical sister. For prophylaxis of GVHD, the patient was treated with cyclosporine and methotrexate. Eighteen days after the transplant, the patient experienced a severe hemolytic transfusion reaction due to an alloantibody (anti‐Jka) produced by donor lymphocytes.

RESULTS: The patient was typed as group A, Jk(a+) before transplantation; the donor was typed as group A, Jk(a–). On Day 18 after transplantation, the immunohematologic screening revealed a positive DAT (C3d 3+) and an alloanti‐Jka. Hemolysis in the patient at that time was indicated by a drop in the Hb and an increase in the LDH level (maximum, 592 IU/L on Day 23).

CONCLUSION: The course of hemolysis and the time of appearance of an alloantibody in this patient meet the criteria for passenger lymphocyte syndrome. In most cases, this syndrome is triggered by ABO system antibodies. This is the first reported case of passenger lymphocyte syndrome after PBPC transplantation that was due to an alloantibody that did not belong to the ABO system.
Immune hemolytic anemia caused by sensitivity to a metabolite of etodolac, a nonsteroidal anti‐inflammatory drug
Authors: Cunha P.D. , Lord R.S. , Johnson S.T. , Wilker P.R. , Aster R.H. , Bougie D.W.
Published: March 2003

BACKGROUND: Immune hemolytic anemia can be caused by sensitivity to many different drugs. In some instances, the sensitizing compound can be identified by in vitro testing, but results are often negative. One reason for this is that a drug metabolite formed in vivo can be the sensitizing agent, but the responsible metabolites have rarely been identified at a chemical level. This report describes a patient who developed severe, Coombs‐positive hemolytic anemia on two occasions after taking the nonsteroidal anti‐inflammatory drug etodolac. Studies were performed to characterize etodolac metabolites to which this patient was sensitive.

CASE REPORT: Serum was tested for antibody in the presence and absence of drug using conventional methods and urine from individuals taking etodolac as a source of drug metabolites. Urinary metabolites of etodolac were identified by high‐pressure liquid chromatography analysis. Glucuronide conjugates of etodolac and the 6‐OH metabolite of etodolac were synthesized in a rat liver microsomal system to obtain reference standards.

RESULTS: The patient's serum gave only trace (±) reactions with normal RBCs in the presence of etodolac but reacted strongly (4+) in the presence of urine from an individual taking this drug. The active urinary metabolites were identified as etodolac glucuronide and 6‐OH etodolac glucuronide.

CONCLUSION: This patient appears to have experienced acute, severe immune hemolytic anemia on two occasions because of sensitivity to the glucuronides of etodolac and 6‐OH etodolac. In patients suspected of having drug‐induced immune hemolytic anemia, RBC‐reactive antibodies can sometimes be detected by using urine from an individual taking the implicated medication as the source of drug metabolites in in vitro reactions. For patients who present with acute immune hemolysis, a careful history of drug exposure should be taken, and, where indicated, confirmatory testing should be performed to identify the sensitizing drug and prevent inadvertent reinduction of hemolysis at a later time.
Evaluation of polyethylene terephthalate for ABO and Rh typing and alloantibody screening
Authors: Anderson D.R. , Wiseman J. , MacLeod J. , Burton E. , Zayed E.
Published: March 2003

BACKGROUND: For many years, hospitals and laboratories have used evacuated glass tubes for blood collection. To improve the safety of blood collection, plastic polyethylene terephthalate (PET) tubes (Vacutainer PLUS, Becton Dickinson) have been developed. The objectives of this study were to compare the accuracy of ABO grouping, Rh typing, and antibody screening of blood samples collected in plastic tubes with that in glass tubes and to determine if refrigerated blood samples collected in plastic tubes remained stable over a 28‐day period.

STUDY DESIGN AND METHODS: Samples were collected from 121 volunteers, at least 30 from each of the A, B, O, and AB blood groups, in four types of Vacutainer tubes: silica‐coated plastic, K2 EDTA plastic, uncoated glass, and K2 EDTA glass. Samples from each tube were tested for ABO group and Rh type by use of the microtyping gel identification card system and the tube method. A three‐cell antibody screen was performed by the microtyping gel card technique with a monospecific IgG reagent. Initial samples were tested within 3 hours of collection. Refrigerated samples were retested for ABO and Rh type and antibody screening 1, 2, 21, and 28 days later. Agreement between test results was determined by using Cohen's Kappa statistic.

RESULTS: Complete agreement was observed between the ABO and Rh typing results in samples drawn into glass and plastic tubes of both the EDTA and nonanticoagulated type (κ = 1.0). In retesting, there were no examples of a change in ABO or Rh type over the 28‐day study period. Only two alloantibodies (1.7%) were identified in the 121 samples, and no difference was observed in alloantibody expression in either plastic or glass Vacutainer tubes over the 28‐day study period.

CONCLUSION: Samples collected into the PET serum or EDTA tubes provided accurate ABO and Rh typing results that remained consistent over a 28‐day period. Samples collected in these tubes also appeared to enable accurate alloantibody identification. However, the number of alloantibodies identified in this study was small, and this result should be confirmed in a larger series.