bluuebunny/crossref_metadata_2025 huggingface dataset:
Mammalian genotoxicity assessment of methylene blue in plasma: implications for virus inactivation
Authors: Wagner S. J. , Cifone M. A. , Murli H. , Dodd R. Y. , Myhr B.
Published: March 2003

BACKGROUND: The risk of adverse consequences of virus‐inactivation procedures for plasma and cellular blood components must be less than the risk of transfusion‐associated viral disease. Previous studies demonstrated that methylene blue, which is currently used in Europe for virus inactivation in fresh‐frozen plasma, can elicit mutations in bacterial test systems. This study investigates the potential for methylene blue genotoxicity in two mammalian test systems.

STUDY DESIGN AND METHODS: Different concentrations of methylene blue were prepared in plasma (heat‐treated at 56 degrees C for 1 hour to reduce cytotoxicity) and used, without illumination, in an in vitro mouse lymphoma cell assay designed to detect forward mutations in the gene encoding thymidine kinase. The assay was performed in the presence or absence of rat liver S9 microsomal fraction. Similarly prepared samples of methylene blue in heat‐treated plasma were used in an in vivo mouse micronucleus assay. Each system included a negative vehicle control (heat‐treated plasma without methylene blue) and a positive control consisting of a known genotoxic agent.

RESULTS: Intravenous administration to mice of 62 mg per kg of methylene blue did not increase the frequency of micronuclei in polychromatic red cells harvested from bone marrow. However, methylene blue concentrations of 10 micrograms per mL (with S9 activation) and 30 micrograms per mL (without S9 activation) significantly increased the thymidine kinase mutation frequency of mouse lymphoma cells to approximately 110 × 10(‐ 6), from a spontaneous frequency of 28 × 10(‐6).

CONCLUSION: Methylene blue is mutagenic in cultured mammalian cells. In contrast, results from the mouse micronucleus assay suggest that the genotoxicity is not expressed in vivo. Considerably more investigation will be required to assess the genotoxic potential of intravenously administered methylene blue used in virus‐inactivation procedures, because of the likelihood of the formation of methylene blue photoproducts or the impact of metabolic conversion of methylene blue to leukomethylene blue in vivo.
Protection of human polymorphonuclear leukocyte function from the deleterious effects of isolation, irradiation, and storage by interferon‐γ and granulocyte‐colony‐stimulating factor
Authors: Rex J. H. , Bhalla S. C. , Cohen D. M. , Hester J. P. , Vartivarian S. E. , Anaissie E. J.
Published: March 2003

BACKGROUND: Fungal infections represent a difficult challenge to clinicians caring for neutropenic patients with hematologic malignancies, as antifungal therapy often has limited success in that setting. One promising yet problematic alternative approach is leukocyte transfusion. The isolation of polymorphonuclear leukocytes (PMNs) induces apoptosis and functional deterioration, and irradiation to prevent transfusion‐associated graft‐versus‐host disease causes further functional deterioration.

STUDY DESIGN AND METHODS: The ability of interferon‐gamma and granulocyte‐colony‐stimulating factor (G‐CSF), used both alone and in combination, to protect PMNs after 0 or 20 hours' storage in cell culture (as a model for function after transfusion) and irradiation with 0, 5, or 30 Gy was studied.

RESULTS: Without cytokine treatment, 20‐hour‐old PMNs showed marked apoptosis, no appreciable chemotaxis, and no ability to kill Candida albicans. In contrast, cytokine treatment significantly reduced apoptosis and protected chemotaxis, C. albicans killing, and surface‐receptor expression from both storage and irradiation. Although the majority of the benefit appeared to be due to G‐CSF, consistent trends suggested better function of PMNs after combined treatment with interferon‐gamma and G‐CSF.

CONCLUSION: Judicious use of cytokines may preserve PMN function. These findings have important implications for the transfusion of PMNs to cytopenic patients.
Anti‐A and/or anti‐B is not detectable in some patients who underwent ABO‐incompatible bone marrow transplantation
Authors: Maeda K. , Taniwaki K. , Santo T. , Itsukuma T. , Ito M. , Anan K. , Misawa M. , Ohe Y. , Kai S. , Hara H.
Published: March 2003

BACKGROUND: Recently, anti‐A and/or anti‐B produced by B cells from donor marrow could not be detected for more than 20 weeks in some patients who had undergone ABO‐incompatible bone marrow transplantation (BMT).

STUDY DESIGN AND METHODS: Twelve to 72 weeks after 11 patients underwent ABO‐incompatible BMT, titers of anti‐A and anti‐B were assayed, A and B antigens were identified by routine methods and flow cytometry, direct and indirect antiglobulin tests were performed, and the red cell antibody was eluted.

RESULTS: In some patients who underwent ABO‐incompatible BMT, anti‐A and/or anti‐B produced by the B cells from the donor marrow could not be detected after BMT when red cells taken from the patients before BMT carried the corresponding antigen‐that is, when hematopoiesis had already changed the cells to the donor's type according to ABO blood typing. Furthermore, some blood samples from those patients gave positive results in direct antiglobulin tests. Blood typing of patients after BMT showed mixed‐ field agglutination. In one patient, the half‐life of red cells assayed with 51Cr was 22.4 days (30.0 +/− 4.0 days for normal controls).

CONCLUSION: Although many hypotheses could be considered to explain the present data, the possibility is proposed that anti‐A and/or anti‐B in the sera must have been consumed in some patients who underwent ABO‐ incompatible BMT. This may lead to problems such as difficulty of ABO typing, positive direct antiglobulin tests, and a relatively short life span of red cells.
Patterns of care for and outcomes of Pneumocystis carinii pneumonia among persons with transfusion‐acquired AIDS
Authors: Bennett C.L. , Horner R.D. , Aboulafia D. , Weinstein R.A.
Published: March 2003

BACKGROUND: The most common human immunodeficiency virus (HIV)‐related cause of death in persons with transfusion‐acquired (TA) AIDS has been Pneumocystis carinii pneumonia (PCP). While better treatment for PCP accounts for improved survival among HIV‐infected homosexual or bisexual men, the extent to which others have benefitted from these developments is unknown.

STUDY DESIGN AND METHODS: Patterns of PCP care among persons with TA‐AIDS, intravenous drug users, and homosexual or bisexual men are compared.

RESULTS: TA‐AIDS patients were older (mean, 46 years vs. < 40 for others, p < 0.05), more severely ill (59% had an alveolar‐arterial oxygen gradient > 48.5 torr vs. 41% of others, p < 0.05), and less likely to have received PCP prophylaxis (16% of TA‐AIDS patients versus 24–41% of others, p < 0.05). PCP care and outcomes also differed: TA‐AIDS patients were less than half as likely to have early use of PCP medications (relative odds ratio = 0.45; 95% CI, 70% vs. > 80% for others, p < 0.05), more likely to be intubated (22% vs. 9–13% of others, p < 0.05), and more likely to die in‐hospital (26% vs. 13‐ 22% of others, p < 0.05). After controlling for differences in severity of illness, insurance, age, and hospital characteristics, TA‐AIDS patients were 45 percent as likely to have early PCP therapy (95% CI, 22%, 91%) as were persons in high‐risk groups.

CONCLUSION: For persons whose only risk factor was transfusion, recognition of the HIV infection and its complications appears to be problematic, which may help explain poorer outcomes in persons with HIV‐related PCP.
A Bombay individual lacking H and Le antigens but expressing normal levels of α‐2‐ and α‐4‐fucosyltransferases
Authors: Shechter Y. , Etzioni A. , Levene C. , Greenwell P.
Published: March 2003

BACKGROUND: The rare Bombay phenotype is usually due to a primary genetic defect in an alpha‐2‐ or alpha‐4‐fucosyltransferase. The present study was done to investigate a patient with normal transferases, who exhibits the Bombay phenotype. CASE REPORT: Red cells of the patient, his parents, and siblings were phenotyped for A, B, and H antigens. The presence of B, H, and Le transferases in serum and saliva was measured.

RESULTS: The parents and siblings were all group B, Le(a‐b‐). The propositus was typed as Oh, Le(a‐b‐). His serum contained anti‐A, anti‐B, and anti‐H. Normal levels of B, H, and Le transferases were found in all family members including the patient.

CONCLUSION: In an unusual case, a person has the Bombay phenotype, but normal levels of transferases in serum and saliva. A general defect in fucose metabolism seems to be the primary abnormality in this case.
Flow cytometric detection of platelet‐reactive antibodies and application in platelet crossmatching
Authors: Kohler M. , Dittmann J. , Legler T.J. , Lynen R. , Humpe A. , Riggert J. , Neumeyer H. , Pies A. , Panzer S. , Mayr W.R.
Published: March 2003

Background: Alloimmunization against HLA or platelet antigens can cause refractoriness to platelet transfusions in multiply transfused patients. Crossmatching of platelet concentrates is effective in overcoming this problem.

Study Design and Methods: A flow cytometric assay was used for simultaneous detection of lymphocyte‐reactive and platelet‐reactive antibodies in a single sample using fluorescein isothiocyanate‐labeled anti‐IgG. This assay was compared with the monoclonal antibody‐specific immobilization of platelet antigens (MAIPA) assay in selected sera containing HLA and platelet antibodies. In a further study, this assay was compared with lymphocytotoxicity test results from thrombocytopenic patients, for whom platelet concentrates were ordered. The results of both assays were then correlated with the 1‐hour corrected count increment, with a corrected count increment greater then 7500 considered as an adequate transfusion response.

Results: The results of the MAIPA and flow cytometric assay in detecting platelet‐reactive antibodies correlated well (p<0.0001, r = 0.84). The sensitivity and specificity of the flow cytometric assay in detecting platelet‐reactive antibodies were 94.7 and 96.3 percent, when the MAIPA assay was taken as a reference. In unselected sera from patients, the sensitivity and specificity of the flow cytometric assays were, respectively, 72.7 and 91.7 percent in detecting lymphocyte‐ reactive antibodies and 70.6 and 77.7 percent in detecting platelet‐ reactive antibodies, when the lymphocytotoxicity test was used as a reference. With regard to an adequate transfusion response, the sensitivities and efficiencies were 20.0 and 82.1 percent, 33.3 and 84.3 percent, and 70.0 and 88.6 percent for the lymphocytotoxicity test and the lymphocyte‐reactive and platelet‐reactive flow cytometric assays, respectively.

Conclusion: Flow cytometric crossmatching appears to be an effective method of detecting platelet‐reactive antibodies that may affect the success of platelet transfusions. This procedure is well‐suited for routine conditions and can be performed within 2 hours.
Genotyping of the human platelet antigen systems 1 through 5 by multiplex polymerase chain reaction and ligation‐based typing
Authors: Legler T. J. , Kohler M. , Mayr W.R. , Panzer S. , Ohto H. , Fischer G. F.
Published: March 2003

BACKGROUND: Platelet‐specific antibodies may be involved in refractoriness to platelet transfusions, disorders such as neonatal alloimmune thrombocytopenia, and post‐transfusion purpura. Genotyping for the major human platelet antigen (HPA) systems HPA‐1 through HPA‐5 is of considerable help in establishing the diagnoses of these diseases.

STUDY DESIGN AND METHODS: A new genotyping method is described. Alleles of all five systems are amplified in a multiplex polymerase chain reaction. Subsequently, aliquots of the amplification products are thermocycled in the presence of a pair of allele‐specific oligonucleotide probes and a heat‐stable ligase. After heat denaturation, the probes hybridize adjacent to complementary sequences of the amplification product. In a perfect match, the two probes become covalently joined. Detection of the ligation product is performed with an enzyme‐linked immunosorbent assay.

RESULTS: Complete concordance of genotypes between the ligation‐based typing and established genotyping methods was determined in 54 Austrian (HPA‐1, −2, −3, and −5) and 56 Japanese (HPA‐4) individuals. Ligation‐based genotyping of HPA‐1 polymorphism using platelet‐derived RNA as starting material gave concordant results in all 15 cases tested.

CONCLUSION: Multiplex polymerase chain reaction in combination with ligation‐based typing allows fast typing of large numbers of platelet donors and screening for critical antigens in pregnant women.
Anti‐Dib as a red cell autoantibody
Authors: Issitt P.D. , Combs M.R. , Allen J. , Melroy‐Carawan H.
Published: March 2003

Background: It is known that some “warm”‐reactive autoantibodies are directed against epitopes on the red cell anion exchanger, protein band 3. Some such antibodies (but not all) recognize Wrb. It is also known that Dia and Dib represent an amino acid polymorphism of band 3.

Study Design and Methods: Autoantibodies from 119 patients were tested against Di(b‐) red cells. Seventy‐four of these autoantibodies were subsequently absorbed with Di(b‐) red cells.

Results: All 119 autoantibodies initially reacted with the Di(b‐) red cells, which showed that none contained only anti‐Dib. Among the 74 adsorbed with Di(b‐) red cells, two were found to contain an unadsorbed anti‐Dib component.

Conclusion: No example of autoanti‐Dib as the only autoantibody present was found among the 119 samples tested. However, 2 (2.7%) of 74 autoantibodies subjected to adsorption with Di(b‐) red cells were seen to contain an anti‐Dib component. This low incidence of autoanti‐Dib is in marked contrast to the high incidence of autoanti‐Wrb, although both antibodies define epitopes associated with the red cell anion exchanger, band 3.
Gene frequencies of human platelet antigens on glycoprotein IIIa in Japanese
Authors: Tanaka S. , Ohnoki S. , Shibata H. , Okubo Y. , Yamaguchi H. , Shibata Y.
Published: March 2003

Background: Polymorphism of glycoprotein IIIa on human platelets is one of the factors in alloimmunization that causes neonatal alloimmune thrombocytopenia and refractoriness to platelet transfusion.

Study Design and Methods: DNA typing methods were originally developed to determine the genotypes of five human platelet antigen (HPA) systems located on glycoprotein IIIa: HPA‐1, HPA‐4, HPA‐6W, HPA‐7W and HPA‐8W. The gene frequencies of these platelet antigens were determined by DNA typing of 331 unrelated Japanese donors.

Results: The gene frequencies of the low‐frequency antigens were 0.002, 0.011, and 0.027 for HPA‐1b, HPA‐4b, and HPA‐6W(b), respectively. All 331 Japanese donors tested were HPA‐7W(a/a) and HPA‐8W(a/a). Moreover, in the present study, none of the donors tested had two or more of these low‐frequency antigens.

Conclusion: The risk of neonatal alloimmune thrombocytopenia and refractoriness to platelet transfusion induced by the antigens of the HPA‐1, HPA‐7W, and HPA‐8W systems was extremely rare in Japanese. However, attention must be paid to the involvement of the HPA‐4 and HPA‐6W systems in these clinical disorders.
DNA typing of recipient blood after massive transfusion
Authors: Wenk R.E. , Chiafari P.A.
Published: March 2003

BACKGROUND: Transfusion is known to alter the recipient's blood group, protein allotype, and alloenzyme phenotype by the introduction of donor material and the dilution of his or her own cells and proteins. There is little information about typing the DNA of blood after transfusion. STUDY DESIGN AND METHODS: Ten adult patients who had undergone massive transfusion with white cell‐containing blood components were studied. Pretransfusion and posttransfusion blood specimen DNA types were compared by using Southern blot and polymerase chain reaction (PCR) analyses. RESULTS: DNA types were identical in all 90 paired observations. CONCLUSION: These preliminary findings, in a limited number of patients, suggest that certain DNA methods may provide reliable typing of adult recipients after massive transfusion.
Determination of the rate of reduction in platelet counts in recipients of hematopoietic stem and progenitor cell transplant: clinical implications for platelet transfusion therapy
Authors: del Rosario M.L. , Kao K.I.
Published: March 2003

BACKGROUND: To investigate how a delay between pretransfusion platelet count measurement and actual platelet transfusion affects the assessment of platelet transfusion responses, the rate of reduction in platelet counts was determined in 30 patients with relatively uncomplicated thrombocytopenia. STUDY DESIGN AND METHODS: Fifteen adult and 15 pediatric patients admitted for hematopoietic stem and progenitor cell transplantation were studied. Platelet counts before and after myeloablative conditioning and after prophylactic platelet transfusions were determined and studied as a function of time. The rates of reduction in platelet counts were determined by linear regression analysis. RESULTS: Platelet counts were reduced at linear rates after myeloablative conditioning or prophylactic platelet transfusion in all 30 patients. The average rates of reduction in platelet counts after myeloablation were 1261 +/− 583 and 1070 +/− 492 platelets per microL per hour (mean +/− SD) for adult and pediatric patients, respectively. The average rate of reduction after platelet transfusions during the thrombocytopenic phase was 740 +/− 280 and 820 +/− 288 platelets per microL per hour (mean +/− SD) for adult and pediatric patients, respectively. The rates of reduction in platelet counts between the two phases were significantly different in the two age groups (adult, p < 0.0001; pediatric, p < 0.015) and were proportionally correlated with initial platelet counts immediately before myeloablation and after prophylactic platelet transfusions. CONCLUSION: The rate of reduction in platelet count can have a significant impact on the evaluation of platelet transfusion responses when there is a delay between pretransfusion measurement of platelet count and the initiation of platelet transfusion. In addition, the rate of platelet reduction determined from this study can be used to confirm an accelerated rate of platelet consumption in thrombocytopenic patients.
Differential effect of donor‐specific blood transfusions after kidney, heart, pancreas, and skin transplantation in major histocompatibility complex‐incompatible rats
Authors: Bektas H. , Jörns A. , Klempnauer J.
Published: March 2003

BACKGROUND: Donor‐specific blood transfusion prior to transplantation has been demonstrated to prolong allograft survival. This study compared the effect of donor‐specific blood transfusion after kidney, heart, pancreas, and skin allotransplantation in congenic rats of reciprocal strain combinations across a major histocompatibility complex barrier. STUDY DESIGN AND METHODS: Whole donor‐specific and third‐party blood was administered to the prospective graft recipients 14 and 7 days before the transplantation of kidney, heart, pancreas, and skin. Rejection was defined by cessation of organ‐specific function (i.e., uremia, cessation of heartbeat, and recurrence of diabetes). The survival of skin grafts was monitored visually. All allografts were histologically assessed immediately after rejection or at the end of a 100‐day observation period. RESULTS: Donor‐specific blood transfusion leads to permanent acceptance of all renal allografts. LEW.1U heart allografts were accepted by LEW.1A recipients, but they were rejected in the reciprocal situation. Survival of pancreas and skin allografts was not significantly prolonged. Third‐party blood had no effect on allograft survival. CONCLUSION: The beneficial effect of donor‐specific blood transfusion depends on both the kind of transplanted organ and the genetic incompatibility involved. The precise mechanisms responsible for the remarkable organ‐specific differences remain unknown.
Evaluation of donor skin disinfection methods
Authors: Goldman M. , Roy G. , Fréchette N. , Décary F. , Massicotte L. , Delage G.
Published: March 2003

BACKGROUND: Because most bacteria isolated from contaminated platelet concentrates are thought to originate from the donor's skin, the efficacy of four methods of skin disinfection was compared.

STUDY DESIGN AND METHODS: Contact plates were used for antecubital skin cultures after they were demonstrated to be easier to use and at least as sensitive as a swab system. One antecubital fossa of each subject was disinfected by a standard method, the use of a povidone‐iodine swabstick containing 0.75‐percent available iodine followed by the use of a povidone‐iodine swabstick containing 1‐percent available iodine. The other arm was disinfected with either a 70‐percent isopropyl alcohol scrub followed by an ampoule of 2‐percent iodine tincture (Group 1; n = 126); a green‐soap sponge followed by a 70‐percent isopropyl alcohol swab, used for donors who are allergic to iodine (Group 2; n = 30); or a 0.5‐percent chlorhexidine gluconate and 70‐percent isopropyl alcohol sponge followed by an ampoule of 0.5‐percent chlorhexidine gluconate and 70‐percent isopropyl alcohol (Group 3; n = 40). Contact plate cultures were done before and after disinfection, and colonies counted after a 48‐hour 37°C incubation period.

RESULTS: Similar numbers of bacteria grew from both antecubital fossae of the same subject before disinfection (p = 0.71). Compared to the standard povidoneiodine method, isopropyl alcohol and tincture of iodine resulted in significantly less bacterial growth (p<0.001), the green soap and isopropyl alcohol method resulted in significantly more bacterial growth (p<0.001), and the chlorhexidine gluconate and isopropyl alcohol method resulted in similar amounts of bacterial growth (p>0.3).

CONCLUSION: Isopropyl alcohol scrub followed by iodine tincture is more efficacious than povidone‐iodine as measured by contact plate cultures. For donors who are allergic to iodine, chlorhexidine gluconate and isopropyl alcohol is more efficacious than green soap and isopropyl alcohol.
Composition of peripheral blood progenitor cell components collected from healthy donors
Authors: Stroncek D. F. , Clay M. E. , Smith J. , Herr G. , Ilstrup S. J. , Kunkel L. A. , McCullough J.
Published: March 2003

BACKGROUND: Peripheral blood progenitor cell (PBPC) components are being collected from healthy donors for allogeneic transplantation, but the quantity, quality, composition, and variability of PBPCs collected from healthy people given granulocyte‐colony‐stimulating factor (G‐CSF) have not been evaluated. STUDY DESIGN AND METHODS: PBPC components were collected from 150 healthy people who were given G‐CSF (5, 7.5, or 10 microg/kg/day) for 5 days. The components were evaluated for white cell (WBC), mononuclear cell, CD34+ cell, neutrophil, platelet, and red cell (RBC) composition. RESULTS: The quantities collected were: WBCs, 35.0 +/− 16.4 × 10(9) (range, 11.9–163.3 × 10(9)); mononuclear cells, 33.3 +/− 14.4 × 10(9) (range, 11.9–139.6 × 10(9)); CD34+ cells, 412 +/− 287 × 10(6) (range, 70–1658 × 10(6)); neutrophils, 1.71 +/− 3.59 × 10(9) (range, 0–27.6 × 10(9)); RBCs, 7.2 +/− 4.0 mL (range, 0–22.1 mL); and platelets, 480 +/− 110 × 10(9) (range, 250–920 × 10(9)). PBPC components collected from people given G‐CSF at 7.5 or 10 microg per kg per day contained significantly more CD34+ cells (respectively, 428 +/− 300 × 10(6); range, 70–1658 × 10(6) and 452 +/− 294 × 10(6); range, 78‐ 1380 × 10(6)) than those from people given G‐CSF at 5 microg per kg per day (276 +/− 186 × 10(6); range, 91–767 × 10(6)) (p = 0.007 and p = 0.002). When 10 microg per kg per day of G‐CSF was given, 50 percent of the components contained enough CD34+ cells for transplantation to a 75‐ kg recipient (375 × 10(6) CD34+ cells), but 10.6 percent of the components contained less than 150 × 10(6) CD34+ cells and thus would provide a transplantable dose only for a 30‐kg patient. CONCLUSION: One PBPC component collected from a healthy donor given 7.5 or 10 microg per kg per day of G‐CSF should contain 70 to 1660 × 10(6) CD34+ cells, with 0 to 22 mL of RBCs. Because of the great variability in the number of CD34+ cells collected, the quantity of CD34+ cells in each component should be measured after each procedure to ensure that sufficient quantities of cells are present for a successful transplant.
Blood group antigens Rb(a), Tr(a), and Wd(a) are located in the third ectoplasmic loop of erythroid band 3
Authors: Jarolim P , Murray J.L. , Rubin H.L. , Smart E. , Moulds J.M.
Published: March 2003

BACKGROUND: Rb(a), Tr(a), and Wd(a) are three low‐incidence blood group antigens that have not been assigned to a particular structure of the red cell membrane. Recent genetic and serologic data suggested erythroid band 3 as a possible carrier of these three antigens. STUDY DESIGN AND METHODS: Ten band 3 gene exons that encode the membrane domain of band 3 were screened for single strand conformation polymorphism (SSCP). Exons displaying SSCP were cloned and sequenced, and the presence of the mutations was verified by restriction digestion. RESULTS: Substitutions 548 Pro–>Leu, 551 Lys–>Asn, and 557 Val–>Met, all located in the third ectoplasmic loop of band 3, were detected in the subjects with Rb(a+), Tr(a+), and Wd(a+) red cells, respectively. The presence of the Rb(a) and Wd(a) mutations was confirmed in additional carriers of these blood group antigens. Chymotryptic cleavage at Tyr 553 and Tyr 555 abolished the agglutinability of Tr(a+) and Wd(a+) cells with the corresponding antisera, further demonstrating that the epitopes are located in the third ectoplasmic loop of band 3. Similar quantities of mRNA corresponding of the two band 3 alleles, a normal pattern of red cell membrane proteins, and normal DIDS (4,4′‐diisothiocyanatostilbene‐2,2′‐ disulphonic acid, disodium salt)‐inhibitable sulfate flux were detected, which suggests that the mutations do not affect band 3 mRNA stability or band 3 protein expression and transport function. CONCLUSION: Wd(a) and Rb(a), and tentatively Tr(a), can be assigned to the Diego blood group system.
The hemostatic effect of packed red cell transfusion in patients with anemia
Authors: Ho C. H.
Published: March 2003

BACKGROUND: The hemostatic effect of platelets has been well established, but the possible role of red cells in hemostasis has not yet been well studied. An evaluation of the hemostatic effect of packed red cell transfusion in patients with chronic anemia was the purpose of this study. STUDY DESIGN AND METHODS: In a prospective study, bleeding time (BT), activated partial thromboplastin time (APTT), and prothrombin time (PT) were measured before and after the transfusion of allogeneic packed red cells in 42 patients with chronic anemia. The results were compared and analyzed. RESULTS: APTT and BT decreased significantly after transfusion, by a mean of 1.3 seconds (p = 0.01) and 2.6 minutes (p < 0.01), respectively. PT did not change significantly after transfusion (p = 0.65). Factors studied (patient's age, sex, and renal function measurements; pretransfusion and posttransfusion hemoglobin levels, platelet counts, and PTs; change in platelet count [delta platelet count] and PT [delta PT] after transfusion) did not independently affect the change in BT (delta BT) or in APTT (delta APTT). The delta BT was not affected by the pretransfusion or posttransfusion levels of APTT or by the delta APTT. The delta APTT was not affected by the pretransfusion or posttransfusion levels of BT or by the delta BT. Diagnosis of malignant or benign diseases was found to affect delta APTT, but not delta BT. Patients with pretransfusion hemoglobin < or = 60 g per L had a 4.07 times greater chance of posttranfusion increase in BT than the patients with hemoglobin > 60 g per L (p = 0.04). CONCLUSION: Red cell transfusion might decrease the APTT and BT in some anemic patients, though the actual cause of the decrease was not determined in the present study.
Retention of “safe” blood donors. The Retrovirus Epidemiology Donor Study
Authors: Thomson R. A. , Bethel J. , Lo A. Y. , Ownby H. E. , Nass C. C. , Williams A. E. ,   Retrovirus Epidemiology Donor Study
Published: March 2003

BACKGROUND: There are obvious advantages to increasing donor retention. However, for reasons of blood safety, certain donors may, in fact, be more desirable to retain than others. “Safe” donors are defined as those who provided a blood donation that was negative on all laboratory screening tests and who subsequently reported no behavioral risks in response to an anonymous survey. This study identifies the most important factors affecting the intention of “safe” donors to provide another donation.

STUDY DESIGN AND METHODS: An anonymous survey asking about donation history, sexual history, injecting drug use, and recent donation experience was mailed to 50,162 randomly selected allogeneic donors (including directed donors) who gave blood from April through July or from October through December 1993 at one of the five United States blood centers participating in the Retrovirus Epidemiology Donor Study. Before mailing, questionnaires were coded to designate donors with nonreactive laboratory screening tests at their most recent donation.

RESULTS: A total of 34,726 donors (69%) responded, with substantially higher response among repeat donors. According to reported intentions only, the vast majority of “safe” donors indicated a high likelihood of donating again within the next 12 months. Only 3.4 percent reported a low likelihood of donating again. A comparison of those likely to return and those unlikely to return reveals significant differences in demographics and in ratings of the donation experience. A higher proportion of those unlikely to return were first‐time donors, minority‐group donors, and donors with less education. The highest projected loss among “safe” donors was seen for those who gave a fair to poor assessment of their treatment by blood center staff or of their physical well‐being during or after donating.

CONCLUSION: These data suggest that efforts to improve donors' perceptions of their donation experience, as well as attention to the physical effects of blood donation, may aid in the retention of both repeat and first‐time donors.
Quality control of white cell‐reduced red cells: white cell preservation and simplified counting
Authors: Tran K. , Nakajo S. , Kapoor K. , Chen M. , Stromberg R.R.
Published: March 2003

BACKGROUND: White cell (WBC) degradation restricts the interval between the filtration process and the assay for residual WBCs. Maintaining WBC integrity would permit extended sample storage for batching and/or shipment to centralized laboratories. The usual quality control assay for WBC‐reduced red cell units requires determining the number of WBCs in the entire counting area of a Nageotte hemocytometer, which consists of 40 rows. Reducing the counting area would simplify the quality control procedure.

STUDY DESIGN AND METHODS: Adsol red cell units were prepared either on the day of collection (Day 0) or on Day 1 and WBC reduced by filtration on the same day. By using prefiltration and postfiltration red cells, samples containing WBC concentrations of 15, 10, and 3 WBCs per microL were prepared by serial dilution. Identical samples were treated with glutaraldehyde and stored at either 20 to 24 degrees C or 1 to 6 degrees C. All samples were assayed on the day of component preparation and on Days 7 and 14. The numbers of WBCs corresponding to 10‐ and 40‐row areas of the Nageotte hemocytometer were determined.

RESULTS: For the conditions and WBC concentration range studied, no significant changes in WBC concentrations were observed through Day 14 for glutaraldehyde‐treated samples stored at either temperature, although there were substantial decreases in untreated samples. A 10‐row measurement was determined to be sufficient for identifying WBC‐reduced red cell units passing the present limit of 5 × 10(6) residual WBCs.

CONCLUSION: Glutaraldehyde treatment can preserve WBCs in red cell samples at least up to Day 14, which provides increased efficiency in quality control for laboratories. Current red cell WBC‐reduction filters produce components that, when assayed, contain fewer than 10 WBCs per full counting area. The simplified procedure would allow reduction of the counting area by 75 percent.
False‐positive eluate reactivity due to the low‐ionic wash solution used with commercial acid‐elution kits
Authors: Leger R. M. , Arndt P. A. , Ciesielski D. J. , Garratty G.
Published: March 2003

BACKGROUND: During the use of commercial red cell (RBC) acid‐elution kits for adsorption and elution (adsorption/elution) studies with anti‐ D, unexpected reactive eluates (anti‐D) were obtained from D‐ RBCs. Such results were not obtained with a parallel xylene method or, historically, with heat and ether methods.

STUDY DESIGN AND METHODS: Single‐donor and commercial polyclonal anti‐D samples were incubated with D+ and D‐ RBCs. Acid eluates were prepared by the manufacturers' directions. Variations in the wash step of the eluate preparation included the use of commercial kit wash solution versus phosphate‐ buffered saline versus solutions of various ionic strengths.

RESULTS: Anti‐D was eluted from 20 of 22 samples of D‐ RBCs after incubation with commercial polyclonal anti‐D (titer 512) and from 2 of 3 samples of D‐ RBCs incubated with single‐donor anti‐D (titer 256). With a low‐ titer (16) single‐donor anti‐D, 0 of 4 eluates from D‐ RBCs reacted. When phosphate‐buffered saline was substituted for the commercial wash solution, 0 of 11 D‐ RBC eluates reacted, as compared with 9 of 11 D‐ RBCs that yielded positive 1+(‐)2+ eluates with the commercial wash solution. If the recommended initial phosphate‐buffered saline wash was omitted before the use of the commercial wash solution, the eluate reactivity was stronger (2+(‐)3+). When low‐ionic‐strength (< 0.03 M) saline was substituted, anti‐D was eluted from D‐ RBCs. All last washes were nonreactive. Antiglobulin tests on all adsorbing D‐ were negative.

CONCLUSION: Commercial wash solutions used for acid elution are at low ionic strength and commonly yield superior eluates, but in the presence of high‐titer antibodies, false‐positive eluates can result. It is our belief that the low‐ionic‐strength wash solution caused aggregation of IgG and nonspecific attachment of IgG on RBCs. Aggregates will contain IgG serum antibodies in proportion to the titer of the antibody. It is this nonspecifically bound antibody that is eluted from antigen‐ negative RBCs.
Perceptions and preferences of autologous blood donors
Authors: Lee S.J. , Liljas B. , Churchill W.H. , Popovsky M.A. , Stowell C.P. , Cannon M.E. , Johannesson M.
Published: March 2003

BACKGROUND:The public's perception of autologous blood donation and transfusion as a worthwhile alternative to allogeneic blood transfusion increased dramatically with discovery of the human immunodeficiency virus. However, new concerns are being raised about the health outcomes and cost‐effectiveness of the procedure. As more restrictive guidelines for autologous blood donation evolve, opposition from patients concerned about exposure to allogeneic blood may arise. Physicians' ability to reassure patients and garner their support for more restrictive policies requires an understanding of patients' concerns. The motivations, perceptions, and preferences of patients currently participating in autologous blood donation programs were investigated in this study.

STUDY DESIGN AND METHODS:Results from two questionnaire studies of 647 autologous blood donors are presented. The questionnaires assessed demographics, risk perceptions, preferences, willingness to pay, and reactions to different interventions designed to decrease patient preference for autologous blood donation.

RESULTS:Patients expressed a strong preference for the availability of autologous blood and indicated that they would be willing to pay substantial amounts of money even if the procedure were not covered by insurance. Despite education about the low risks of complications from allogeneic transfusions, an aversion to allogeneic transfusion and a willingness to pay for autologous blood donation persisted. Patients were not reassured by information on better infectious disease testing or physician recommendation against autologous blood donation. CONCLUSION: Patients currently participating in autologous blood donor programs strongly prefer continued access to this procedure, primarily because they remain concerned about the complications of allogeneic transfusions. They may not be significantly reassured despite increasingly rigorous and costly improvements in donor and component screening.
High prevalence in Central Africa of blood donors who are potentially infectious for human herpesvirus 8
Authors: Bélec L. , Cancré N. , Hallouin M.‐C. , Morvan J. , Mohamed A. Si , Grésenguet G.
Published: March 2003

BACKGROUND: In western countries, the transmission of human herpesvirus 8 (HHV‐8) via blood transfusion has been recently postulated. In sub‐ Saharan African, the incidence of HHV‐8‐associated Kaposi's sarcoma and the seroprevalence for HHV‐8 in autochthonous populations are high.

STUDY DESIGN AND METHODS:The aim of this study was to estimate the prevalence of blood donations potentially infectious for HHV‐8 in the general adult population of Central Africa. Forty‐nine blood donors at the Centre de Transfusion Sanguine in Bangui, the capital of the Central African Republic, were selected. Forty‐five inpatients of Broussais Hospital, Paris, France, who were known to be seronegative for HIV and hepatitis B and C viruses and who had not received heart or kidney transplants, were chosen as a European “control” group for comparison. HHV‐8 DNA sequences were detected in peripheral blood mononuclear cells by nested polymerase chain reaction using primer sets located in the HHV‐8 open reading frame 26.

RESULTS:Eleven (22.5%; 95% CI: 12%‐37%) of 49 blood donors were positive for HHV‐8. Three (6%) were HIV‐1 seropositive. Two (67%) of the 3 HIV‐infected blood donors were also positive for HHV‐8. All blood donors were apparently healthy; none was known to suffer from Kaposi's sarcoma. Only one (2.2%) control was carrying HHV‐8 DNA on the peripheral blood mononuclear cells. The prevalence of HHV‐8 was higher in blood donors from Bangui than in patients from Broussais Hospital.

CONCLUSIONS:HHV‐8 infection is highly prevalent in an apparently healthy adult population from Central Africa, which raises concerns about HHV‐8 transmission through transfusion in this setting.
Anti‐G in a pregnant patient
Authors: Cash K. , Brown T. , Strupp A. , Uehlinger J.
Published: March 2003

BACKGROUND: Anti‐G is a red cell (RBC) antibody of the Rh system. It has been described in pregnant women only in association with anti‐D or anti‐C; therefore, the ability of this antibody alone to cause hemolytic disease of the newborn is uncertain. One case in which this antibody caused no clinical sequelae is reported.

CASE REPORT: The patient was a 35‐year‐old primigravida with type O, Dñ, Cñ, Eñ, c+ RBCs who was given 4 units of type O, Dñ allogeneic RBCs and 2 units of autologous RBCs 2 years antepartum. She was found to have anti‐D and anti‐C by an outside laboratory as part of a routine prenatal work‐up. Further evaluation by our laboratory revealed the presence of anti‐G and possible anti‐C without anti‐D. Titers at 22 weeksí gestation were 64 against r′r RBCs and 16 against R2R2 RBCs; these remained unchanged throughout the pregnancy. Amniocentesis performed at Weeks 28 and 32 showed no evidence of hemolytic disease of the newborn. A healthy 3.3‐kg infant was delivered at 36 weeksí gestation. Prophylactic Rh immune globulin was administered antepartum and postpartum. The infantís RBCs were type O, D+, c+ Cñ, Eñ, and the direct antiglobulin test was positive. An acid eluate prepared from the babyís RBCs revealed anti‐G. The total bilirubin was 5.5 mg per dL at birth, and the hematocrit was 66 percent. Total bilirubin peaked on Day 5 at 11.9 mg per dL, and no therapeutic intervention was required.

CONCLUSIONS: Anti‐G alone caused little if any fetal or neonatal hemolysis in this case. Although further study is needed, invasive fetal monitoring may be unnecessary if anti‐G is the sole cause of fetomaternal RBC incompatibility.
Delayed massive immune hemolysis mediated by minor ABO incompatibility after allogeneic peripheral blood progenitor cell transplantation
Authors: Salmon J.‐P. , Michaux S. , Hermanne J.‐P. , Baudoux E. , Gérard C. , Sontag‐Thull D. , Fillet G. , Beguin Y.
Published: March 2003

BACKGROUND: Bone marrow transplantation with minor ABO incompatibility may be followed by moderate delayed hemolysis of the recipient's red cells by donor‐derived ABO antibodies. This reaction may be more severe after transplantation of peripheral blood progenitor cells (PBPCs).

CASE REPORT: A 16‐year‐old boy underwent an allogeneic PBPC transplant from his HLA‐mismatched mother as treatment for acute myeloblastic leukemia that had proved resistant to induction chemotherapy. Transfusion of the unmanipulated PBPCs proceeded without any complication, despite the difference in ABO blood group (donor, O Rh‐positive; recipient, A Rh‐positive). On Day 7, a rapid drop in hemoglobin to 4 g per dL was observed, which was attributed to a massive hemolysis. All the recipient's group A red cells were destroyed within 36 hours. This delayed and rapidly progressive hemolytic anemia was not associated with the transfusion of the donor's plasma. Rather, the anti‐A titer increased in parallel with marrow recovery, which suggested an active synthesis of these antibodies by immunocompetent cells from the donor against the recipient's red cells. The mother's anti‐A titer was retrospectively found to be 2048. Her unusually high titer is probably due to prior sensitization during pregnancies. On Day 12, the patient developed grade IV graft‐versus‐host disease, which proved resistant to all treatments instituted and led to his death on Day 35.

CONCLUSION: PBPC transplantation with minor ABO incompatibility may be associated with significant risk of massive delayed hemolysis.
First two cases of immune hemolytic anemia associated with ceftizoxime
Authors: Shammo J.M. , Calhoun B. , Mauer A.M. , Hoffman P.C. , Baron J.M. , Baron B.W.
Published: March 2003

BACKGROUND: Second‐ and third‐generation cephalosporins have been associated with immune‐mediated hemolytic reactions. This report discusses two patients who developed clinically significant extravascular hemolysis while receiving the third‐generation cephalosporin ceftizoxime (Ceftizox). This is believed to be the first time hemolysis has been described in patients receiving this drug.

STUDY DESIGN AND METHODS: Immunologic workup of drug‐dependent antibodies was performed on blood samples using drug‐coated and immune complex methodologies. Antibody classes and titers were analyzed.

RESULTS: Both the patients' sera contained anti‐ceftizoxime that reacted with red cells only when ceftizoxime was added to the sera (“immune complex” method). The patients recovered without complications following discontinuation of the drug. Each patient had IgM and IgG drug‐dependent antibodies. The drug‐induced antibodies from each patient cross‐reacted with cefotaxime, which is structurally similar to ceftizoxime, but cross‐reacted either weakly or not at all with ceftriaxone, which has a more complex side chain.

CONCLUSION: This report describes the first cases of immune hemolytic anemia associated with ceftizoxime. In drug‐induced hemolytic reactions, prompt recognition and discontinuation of the drug may be important factors in reducing the chance of serious sequelae.
Improved removal of white cells with minimal platelet loss by filtration of apheresis platelets during collection
Authors: Holme Stein , Andres Mark , Goermar Nancy B. , Giordano Gerald F.
Published: March 2003

BACKGROUND: Filtration of apheresis platelets to remove white cells (WBCs) requires operator intervention after the collection procedure (postcollection filtration), which may cause variable and unsatisfactory filter performance (WBC removal and platelet loss). The MCS+ LN9000 apheresis system filters platelets through a WBC‐reduction filter during each collection cycle (continuous filtration) at a flow rate of 15 to 25 mL per minute. Apheresis platelets obtained by continuous filtration were evaluated in terms of platelet loss, WBC removal, and platelet storage properties and then were compared to unfiltered apheresis platelets and to apheresis platelets that underwent postcollection filtration. Two WBC‐reduction filters were tested (LRF6 and LRFXL).

STUDY DESIGN AND METHODS: In 70 apheresis platelets, postcollection filtration was performed by using the LRF6 at flow rates of 80 mL per minute (n = 30) and 50 mL per minute (n = 30) and the LRFXL at 50 mL per minute (n = 10). One hundred fifty‐eight apheresis platelets underwent continuous filtration through the LRF6 (n = 58) or the LRFXL (n = 100). Unfiltered apheresis platelets (controls) (n = 30) were obtained by the same collection protocol.

RESULTS: Estimated platelet loss with continuous filtration was 7 percent for the LRFXL and 3 percent for the LRF6. A reduction in the filtration flow rate from 80 to 50 mL per minute with postcollection filtration through the LRF6 resulted in markedly lower WBC levels, with 10 percent versus 57 percent of the apheresis platelets having WBC counts <1 × 105, respectively. Additional improvements in WBC removal were found with continuous filtration; 85 percent of the apheresis platelets filtered with the LRF6 and 100 percent of the apheresis platelets filtered with the LRFXL had WBC counts <1 × 105.

CONCLUSIONS: Continuous or postcollection filtration of freshly collected apheresis platelets resulted in minimal platelet loss. Better WBC removal from apheresis platelets was obtained with continuous filtration than with postcollection filtration, likely because of the slower flow rate. Platelet storage quality was not affected by filtration.
Prevalence of GB virus type C/hepatitis G virus RNA and of anti‐E2 in individuals at high or low risk for blood‐borne or sexually transmitted viruses: evidence of sexual and parenteral transmission
Authors: Lefrère Jean‐Jacques , Roudot‐Thoraval Françoise , Morand‐Joubert Laurence , Brossard Yvon , Parnet‐Mathieu Françoise , Mariotti Martine , Agis Franz , Rouet François , Lerable Joelle , Lefèvre Guillaume , Girot Robert , Loiseau Pascale
Published: March 2003

BACKGROUND: The first epidemiologic evidence of GB virus type C (GBV‐C)/hepatitis G virus (HGV) infection showed a high prevalence of asymptomatic carriers in blood donors and in populations at risk for blood‐borne viruses. However, by using only viral RNA polymerase chain reaction, those studies underestimated the true spread of GBV‐C/HGV infection. The combined detection of GBV‐C/HGV RNA and of anti‐E2 (which reflects recovery from infection) is necessary to define accurately the prevalence of GBV‐C/HGV.

STUDY DESIGN AND METHODS: The presence of both anti‐E2 and GBV‐C/HGV RNA was searched for in 1438 serum samples collected from various groups of individuals at low or high risk for blood‐borne or sexually transmitted viruses (blood donors, organ donors, unselected pregnant women, immunocompetent or immunodepressed multiply transfused patients, HIV‐positive or HIV‐negative homosexual men, intravenous drug addicts).

RESULTS: The presence of GBV‐C/HGV RNA and/or anti‐E2 (exposure to GBV‐C/HGV) was frequent in populations at risk for blood‐borne or sexually transmitted viruses. GBV‐C/HGV appeared also to be sexually transmitted, with transmission from male to female more efficient than vice versa. A particularly elevated level of exposure to GBV‐C/HGV was observed in homosexual men. In immunocompetent individuals, the prevalence of anti‐E2 was about twice that of GBV‐C/HGV RNA, which suggests the frequency of recovery from GBV‐C/HGV infection. Most of the GBV‐C/HGV RNA‐positive individuals had no biochemical evidence of liver damage.

CONCLUSIONS: GBV‐C/HGV is frequent in populations at risk for blood‐borne or sexually transmitted viruses. GBV‐C/HGV is not a hepatitis virus, and it seems appropriate to rename it.