bluuebunny/crossref_metadata_2025 huggingface dataset:
A prospective microbiologic surveillance program to detect and prevent the transfusion of bacterially contaminated platelets
Authors: Yomtovian R. , Lazarus H.M. , Goodnough L.T. , Hirschler N.V. , Morrissey A.M. , Jacobs M.R.
Published: March 2003

After two patients received bacterially contaminated platelet transfusions, a prospective surveillance program was instituted to perform Gram staining and microbiologic culturing of platelets at the time of transfusion. In 12 months, 3141 random‐donor platelet pools (prepared from 14,481 units) and 2476 single‐donor apheresis units were cultured. All single‐donor apheresis units were sterile, but 6 (0.19%) of the random‐donor pools were found to be bacterially contaminated, with 1 unit of 5 in the pool being the source in each case. Contaminants were Staphylococcus epidermidis (4 cases), Bacillus cereus (1), and Staphylococcus aureus (1) at counts of 0.5 × 10(2) to 10(11) colony‐forming units per mL in platelet pools and 10(3) to 10(13) colony‐forming units per mL in source units. The contamination rate for units transfused at < or = 4 days (1.8/10,000) was significantly lower than that at 5 days (11.9/10,000; p < 0.05), as was the magnitude of contamination (p < 0.05). Use of the pretransfusion Gram stain on 4‐ and 5‐day‐old platelet pools was 100 percent sensitive (4/4 true positives) and 99.93 percent specific (1 false positive) in detecting contaminated pools. These data define the extent and magnitude of platelet bacterial contamination and demonstrate the efficacy of the pretransfusion Gram stain on platelet units stored for 4 and 5 days in preventing the transfusion of heavily contaminated units. It is concluded that the risk of platelet contamination is related to the duration of component storage. Transfusion services can minimize this risk by selecting shorter platelet outdates or by screening 4‐ and 5‐day‐old platelets with a pretransfusion Gram stain.
Complement activation in stored platelet concentrates
Authors: Miletic V. D. , Popovic O.
Published: March 2003

Activation of platelets during preparation and/or storage of platelet concentrates in plastic containers at room temperature has recently been recognized. Many different biologic causes of this activation have been postulated. Activated complement, as a multi‐enzyme system, is one of the possible sources of molecules leading to platelet activation. To detect complement activation, functional complement activity and the generation of complement‐derived ligands were investigated in platelet concentrate supernatant plasma during 5 days of storage at room temperature. Hemolytic tests for functional classical and alternative pathway activity were used, as was the kinetic test for complement‐ mediated inhibition of immune complex precipitation. The presence of C3 activation products (C3, C3c, C3dg) was investigated in plasma by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and Western blotting procedures and on platelets by immunofluorescence. Activation of complement was evident during storage, and C3c and C3d fragments were clearly demonstrated in plasma. The amount of C3d fragments on platelets gradually rose during the first 3 days of storage. At the end of 5 days of storage, the platelets became C3d negative. There are two possible mechanisms of C3d disappearance‐shedding and/or further degradation of C3d fragments. Those results indicated that complement activation and the generation of complement‐dependent ligand‐receptor interaction may be mechanisms for platelet activation in concentrates stored at room temperature.
Identification of the blood component responsible for increased susceptibility to gut‐derived infection
Authors: Gianotti L. , Pyles T. , Alexander J.W. , Fukushima R. , Babcock G.F.
Published: March 2003

It has previously been reported that the transfusion of allogeneic whole blood increases sepsis‐related mortality and decreases the ability of the host to kill bacteria that have translocated from the intestinal tract. To determine which blood component contributes to this adverse effect, the impact of the transfusion of white cells (WBCs), red cells (RBCs), and plasma on microbial translocation, bacteria killing, and mortality rate was studied. Blood from C3H/HeJ mice was separated into WBCs, RBCs, and plasma, and these fractions were transfused to Balb/c mice. Controls received sterile saline. Five days after transfusion, all Balb/c mice underwent a 20‐percent burn and gavage with 1 × 1010 Escherichia coli labeled with 14C‐glucose. Mortality was observed for 10 days. Four additional groups, receiving the same treatment as above, were sacrificed 4 hours after the burn, and mesenteric lymph nodes, liver, kidney, and blood were harvested aseptically. For each tissue, quantitative colony counts, radionuclide counts, and percentage of translocated bacteria that remained alive were calculated. By radionuclide counts, no difference was observed in the degree of 14C E. coli translocation among the groups. In contrast, the percentage of viable bacteria and the mortality rate were significantly higher in the group receiving allogeneic WBCs than in all other groups (p < 0.05). It is concluded that WBCs are the component in transfused blood that has an adverse effect on host resistance to gut‐derived infection.
Survival and half‐life of red cells salvaged after hip and knee replacement surgery
Authors: Umlas J. , Jacobson M.S. , Kevy S.V.
Published: March 2003

The survival of autologous red cells collected intraoperatively has been reported previously. This study measures the survival and half‐ life of red cells collected 3 hours after hip and knee arthroplasty. For six patients, four having knee replacements and two having hip replacements, the salvaged red cells were labeled with radioactive 51Cr. Peripheral blood was simultaneously labeled with nonradioactive 52Cr. There was no significant difference in the survival or half‐life of the salvaged and the venous blood.
Transfusion‐associated graft‐versus‐host disease in immunocompetent patients: report of a fatal case associated with transfusion of blood from a second‐degree relative, and a survey of predisposing factors
Authors: Petz L.D. , Calhoun L. , Yam P. , Cecka M. , Schiller G. , Faitlowicz A.R. , Herron R. , Sayah D. , Wallace R.B. , Belldegrun A.
Published: March 2003

A patient without evident immune deficiency who received a transfusion of blood from a second‐degree family member developed fatal transfusion‐ associated graft‐versus‐host disease (TA‐GVHD). The donor was homozygous for an HLA haplotype for which the recipient was heterozygous (one‐way HLA match). All 39 reported cases of TA‐GVHD in immunocompetent patients were reviewed to ascertain the predisposing factors and to define the indications for irradiating blood for this population. HLA typing was described in 15 cases; in 13, including seven related and six unrelated donors, a one‐way HLA match was present. Thirty‐one (79%) of the 39 cases were reported from Japan (and 196 other cases are cited in the Japanese literature), but a one‐way HLA match among unrelated donors at HLA‐A, ‐B, ‐DR loci is only approximately two to four times more likely in Japanese persons than in whites. Fresh blood (< 96 hours old) was used in 29 (94%) of the 31 cases reported from Japan and in 33 (87%) of 38 cases overall (in one case, the age of the blood used was not reported). Thus, factors that appear to predispose to TA‐GVHD in immunocompetent patients are a one‐ way HLA match, fresh blood, and, possibly, Japanese ancestry. Irradiating cellular blood components from all blood relatives of transfusion recipients will not completely eliminate the risk of TA‐ GVHD.
Duplication of exon 3 in the glycophorin C gene gives rise to the Lsa blood group antigen
Authors: Reid M. E. , Mawby W. , King M.‐J. , Sistonen P.
Published: March 2003

BACKGROUND: The Gerbich‐related Lsa blood group antigen (Ge6) resides on the higher‐molecular‐weight forms of glycophorin C (GPC) and glycophorin D (GPD). Southern blot analysis has previously revealed an additional GPC exon 3 insert in the genomic DNA from an Ls(a+) individual.

STUDY DESIGN AND METHODS: To confirm the duplication of exon 3 in the GPC mRNA, total RNA prepared from the Epstein‐Barr virus‐ transformed lymphocytes of an Ls(a+) individual was used in the synthesis of first‐strand cDNA. The first‐strand cDNA served as a template for the amplification of GPC‐related DNA by polymerase chain reaction. After subcloning, the polymerase chain reaction cDNA was sequenced with a kit. Hemagglutination inhibition of anti‐Lsa sera with synthetic peptides was performed to identify the location of Lsa on the GPC.Lsa protein.

RESULTS: Sequencing the GPC.Lsa cDNA showed an insert of 84 nucleotides, which corresponds to the entire sequence of exon 3 in the GPC gene (GYPC). Since the exon 3‐duplicated exon 3 boundary of GYPC.Lsa encodes a novel amino acid sequence on GPC.Lsa and GPD.Lsa, synthetic peptides consisting of the amino acids flanking this junction were used to determine the amino acid sequence that is essential for expression of Lsa. The peptide DIVVIA/EPDPG was noninhibitory, while peptides with the sequence TPTIMDIVVIA/EPDPG or SPSVLDIVVIA/EPDPG inhibited anti‐Lsa reactivity.

CONCLUSION: Lsa is located within the sequence of amino acids encoded by the nucleotides at the exon 3‐ duplicated exon 3 boundary, and the conformation of this peptide sequence is important for recognition of Lsa by human anti‐Lsa.
Concurrent comparison of the safety of paid cytapheresis and volunteer whole‐blood donors
Authors: Strauss R.G. , Ludwig G.A. , Smith M.V. , Villhauer P.J. , Randels M.J. , Smith‐Floss A. , Koerner T.A.
Published: March 2003

Background: Historically, paid blood donors were found to transmit hepatitis at higher rates than volunteers. In those older studies, paid donors frequently were recruited from prisons or slum areas–a finding consistent with the belief that monetary payment in itself did not necessarily lead to the high‐risk status of commercial blood. Instead, it was the population base from which the donors were recruited that was important.

Study Design and Methods: Today, cytapheresis donors are in great demand. Because payment is one incentive that might entice donors to undertake the increased commitment of repeated cytapheresis donation, the results were studied of infectious disease history and laboratory testing performed concurrently in 917 volunteer whole‐blood donors and 1240 paid cytapheresis donors, who were enrolled in distinct programs at the DeGowin Blood Center from October 7, 1987, through November 30, 1990.

Results: When first, repeat, and overall donations made by these donors were evaluated separately, paid cytapheresis donors were found to exhibit no increase in infectious disease history or test results beyond those of volunteer whole‐blood donors.

Conclusion: Thus, paid cytapheresis donors, when managed within a formal program, should not necessarily be presumed to be more dangerous than volunteers, from an infectious disease aspect. However, definitive proof of safety (comparison of transfusion‐transmitted infection rates in two groups of patients receiving blood components exclusively from either paid cytapheresis or volunteer donors) was not pursued by long‐ term follow‐up studies.
Long‐term follow‐up testing of red cell alloantibodies
Authors: Ramsey G. , Smietana S.J.
Published: March 2003

Background: In previous studies, 29 to 34 percent of potentially hemolytic red cell antibodies were not detected after short‐term follow‐ up.

Study Design and Methods: To examine long‐term detection, records were reviewed for 44 consecutive patients who were tested more than 5 years after their potentially hemolytic red cell antibodies were first identified in this hospital.

Results: After 5 to 10 years, 14 (39%) of 36 Rh, Kell, and Duffy system antibodies were not detected on at least one occasion. Twenty‐two other such antibodies were sought again after more than 10 years; 10 (45%) were not detected. When restimulation by pregnancy was excluded, these rates were 42 and 48 percent, respectively.

Conclusion: Clinically significant red cell antibody formation is probably more common than previously realized, because nearly half of these antibodies are undetected after long‐term follow‐ up.
Independent roles for platelet crossmatching and HLA in the selection of platelets for alloimmunized patients
Authors: Friedberg R.C. , Donnelly S.F. , Mintz P.D.
Published: March 2003

Background: Although HLA‐matched platelets are frequently requested for alloimmunized patients, recent evidence has indicated that 1‐hour posttransfusion platelet increments in these patients are specifically sensitive to crossmatch compatibility.

Study Design and Methods: To determine the extent of advantage gained by use of single‐donor apheresis (SD) platelets selected on the basis of HLA match when crossmatch‐compatible SD platelets were available, a total of 220 platelet transfusions given in the absence of individually determined significant nonimmune factors were analyzed in a well‐characterized cohort of platelet‐refractory patients. Platelets were selected by solid‐phase crossmatch from a small donor pool of relatively poor HLA matches or, upon request, ordered as HLA‐matched and later crossmatched.

Results: Alloimmunized patients responded better to SD platelets selected on the basis of HLA than to pooled platelet concentrates or SD platelets selected at random, although most of the benefit was limited to the 57‐percent subset of good HLA matches. Crossmatch‐compatible SD platelets provided similar posttransfusion platelet increments independent of the HLA match. None of 31 crossmatch‐ incompatible SD platelets transfused provided an adequate increment, including 13 that were ordered as HLA‐matched platelets.

Conclusion: No benefit could be demonstrated from requesting that SD platelets be HLA‐ matched when crossmatch‐compatible SD platelets were available.
Sulindac‐induced immune hemolytic anemia
Authors: Angeles M.L. , Reid M.E. , Yacob U.A. , Cash K.L. , Fetten J.V.
Published: March 2003

Background: Sulindac, a nonsteroidal, anti‐inflammatory, indene‐derived drug, caused life‐threatening immune hemolytic anemia in an individual with back pain.

Case Report: A patient was admitted to the hospital with immune hemolytic anemia and kidney and liver failure after several days ingestion of sulindac. The direct antiglobulin test was positive with polyspecific and monospecific anti‐IgG but not with anti‐C3. The eluate did not react in routine tests but reacted strongly after the addition of sulindac. The serum contained a sulindac‐dependent antibody reacting to a titer of 32. The sulindac‐dependent antibody was of both IgG and IgM classes and had no apparent blood group specificity. The antibody agglutinated red cells from humans and chimpanzees but not from chickens, rabbits, or sheep, which implied that a specific component on human and chimpanzee red cells was needed for reactivity. The antibody reacted with red cells treated with trypsin, papain, pronase, dithiothreitol, and sialidase. With aggressive medical care, the patient's condition improved.

Conclusion: These findings appear compatible with the so‐called immune complex mechanism for drug‐induced immune hemolytic anemia. Physicians are alerted to the severe nature of this syndrome.
The effect of an intraoperative treatment algorithm on physicians' transfusion practice in cardiac surgery
Authors: Despotis G.J. , Grishaber J.E. , Goodnough L.T.
Published: March 2003

Background: Inappropriate transfusion in cardiac surgery may, in part, be due to empiric transfusion therapy instituted in the absence of timely laboratory data. Therefore, the effect of a transfusion decision algorithm based on intraoperative coagulation monitoring of physicians' transfusion practice and the transfusion outcome was evaluated.

Study Design and Methods: In a randomized, controlled trial, cardiac surgical patients determined to have microvascular bleeding at the cessation of cardiopulmonary bypass were assigned to algorithm (A) or standard (S) therapy. Group A was treated with plasma and platelet therapy according to a transfusion algorithm based on on‐site coagulation data available within 4 minutes. For Group S, the use of laboratory‐based data and the decision to transfuse blood components were at physician discretion.

Results: Sixty‐six patients were entered into the study (Group A, n = 30; Group S, n = 36). Other than the fact that there were significantly more female patients in Group S than in Group A, no differences between cohorts in regard to perioperative risk factors for blood transfusion needs were identified. Therefore, gender was factored in as a covariate in the statistical analysis. Group A patients received fewer hemostatic blood component units (p = 0.008) and had fewer total donor exposures (p = 0.007) during the entire hospitalization period. Linear regression analysis of the differences in slopes in Groups A and S for the relationships between the red cell volume lost and the red cell volume transfused (p < 0.03), non‐red cell units transfused (p < 0.0001), and total number of blood components transfused (p < 0.0001) demonstrated that physicians' transfusion practice was significantly altered by the use of a transfusion algorithm with on‐site coagulation data, independent of surgical blood losses.

Conclusion: The use of algorithms by transfusion decision makers can serve as an effective physician education intervention.
Long‐term survival after blood transfusion
Authors: Vamvakas E.C. , Taswell H.F.
Published: March 2003

Background: Look‐back investigations of populations of patients admitted to major tertiary‐care hospitals in the 1980s found a 2‐year posttransfusion mortality rate in excess of 50 percent. To quantify the association of blood transfusion with mortality in a more broadly based population, a cohort of all residents of a United States county who underwent transfusion in 1981 was studied.

Study Design and Methods: Retrospective cohort study comprised 802 county residents. Complete follow‐up (until death or for 10 years) was available on 93.9 percent.

Results: The median length of survival was 95.0 (± 2.5) months. Twenty‐four percent of patients died within 1 year after the transfusion, 30 percent within 2 years, 40 percent within 5 years, and 52 percent within 10 years. The relative risk of death within 10 years increased by 4.1 percent per unit of red cells (p < 0.0001), by 1.2 percent per unit of platelets (p = 0.0003), and by 7.3 percent per unit of fresh‐frozen plasma (p = 0.0018) received in 1981, after adjustment for the effects on mortality of age, gender, and number of days of hospitalization in 1981.

Conclusion: Receipt of a blood transfusion can be used as a descriptive epidemiologic index of morbidity in the general population, as it is independently predictive of mortality, adding to the predictive value of age, gender, and previous hospitalization. There is a dose‐response relationship between the amount of blood components received and a reduction in the subsequent length of survival. However, when a county's entire population is studied, posttransfusion mortality due to underlying disease is substantially lower than that previously reported in look‐back investigations.
Aprotinin efficacy on intraoperative bleeding and transfusion requirements in orthotopic liver transplantation
Authors: Patrassi G.M. , Viero M. , Sartori M.T. , De Silvestro G. , Rossaro L. , Burra P. , Nolli M.L. , Piccinni P. , Bassi N.
Published: March 2003

Background: Bleeding complications frequently occur during orthotopic liver transplantation (OLT), particularly in patients with liver cirrhosis. Enhanced fibrinolytic activity in plasma was seen to play a key role in the development of the hemostatic disorder and of hemorrhages. Aprotinin, a serine protease inhibitor, has been used in the prevention and/or treatment of hyperfibrinolytic states.

Study Design and Methods: In the present study, the effect of aprotinin on bleeding complications and transfusion requirements was investigated in OLT patients with liver cirrhosis. Seven consecutive cirrhotic patients undergoing OLT were infused with aprotinin following an original protocol (1,000,000‐KIU intravenous loading dose plus 500,000 kallikrein‐inhibitory units per hour until skin closure). Seven previous cirrhotic OLT patients not receiving aprotinin were used as controls.

Results: In the treated group, a significant decrease in the number of transfused units of packed red cells (48.7%, p < 0.01), fresh‐ frozen plasma (24.4%, p < 0.05), platelets (35.9%, p < 0.01), and autologous blood (55.2%, p < 0.01) was observed as compared with the control group. Moreover, the mean length of operation was significantly shorter in the aprotinin‐infused patients than in untreated patients (8.3 ± 1.2 vs. 10.1 ± 1.8 hours, respectively; p < 0.01)). In the aprotinin‐treated group, the antifibrinolytic efficacy was confirmed by the lack of increase in D‐dimer levels and decrease of fibrinogen in plasma; on the contrary, these changes were always seen in the group not receiving aprotinin.

Conclusion: Infusion of aprotinin during OLT in cirrhotic patients can be recommended for the prevention of hyperfibrinolysis‐triggered bleeding, thus reducing transfusion requirements. A possible protective effect on the primary nonfunction of the grafted liver is suggested.
Hepatitis C virus among blood donors: follow‐up study
Authors: Alonso C. , Pedroso M.L. , de Sanjosé S. , Montcharmont P. , Chèvre J.M. , Boucaud M.J. , Lambert V. , Cortey M.L. , Trépo C.
Published: March 2003

Background: The exact significance of antibodies to hepatitis C virus (HCV) in blood donors remains unknown. Confirmatory tests of anti‐HCV‐ reactive serum and HCV RNA by polymerase chain reaction (PCR) are used to refute a large proportion of false‐positive results.

Study Design and Methods: Ninety‐two blood donors who were anti‐HCV reactive in a first‐generation enzyme‐linked immunosorbent assay (ELISA) were reevaluated 10 months later with a second‐generation ELISA (ELISA‐2) as well as with second‐generation recombinant immunoblot assay (RIBA‐2) and by PCR.

Results: Twenty‐five (43.9%) of the 57 ELISA‐2‐positive donors were confirmed as positive by RIBA‐2; of these, 84 percent were HCV RNA positive in PCR. Of the 57 who were still anti‐HCV positive, 46 were followed up and tested again in the same manner 2 years after the first screening. At that time, the pattern was little changed: 94 percent of RIBA‐2‐ and PCR‐positive donors remained positive. Of RIBA‐2‐ and PCR‐positive blood donors, 62 percent had abnormal alanine aminotransferase levels in at least one of the three evaluations. Among the anti‐HCV‐positive donors confirmed by RIBA‐2, 60 percent, versus 12.6 percent in the control group, had a significantly (p < 0.001) more frequent risk factor for HCV infection, due to parenteral exposure to blood.

Conclusion: These data confirm a good correlation between RIBA‐2 reactivity and the detection of HCV RNA in a population of anti‐HCV‐ positive blood donors.
Inactivation of free and cell‐associated human immunodeficiency virus in platelet suspensions by aminomethyltrimethylpsoralen and ultraviolet light
Authors: BENADE L.E. , SHUMAKER J. , XU Y. , CHEN X. , DoDD R.Y.
Published: March 2003

BACKGROUND: It has previously been reported that 40 micrograms per mL of aminomethyltrimethylpsoralen (AMT) plus 2.4 to 7.2 J per cm2 of ultraviolet A (UVA) light inactivated 4 to 6 log10 of several model viruses in platelet suspensions. This inactivation was achieved while satisfactory levels of platelet count, pH, morphology, aggregation, and hemostatic effectiveness were maintained.

STUDY DESIGN AND METHODS: The efficacy of this procedure for inactivating free and intracellular human immunodeficiency virus (HIV), including integrated proviral sequences, was studied. RESULTS: The kinetics of inactivation for free HIV (4–5 log10 kill with 1.2–4.8 J/cm2) were similar to those obtained for the previously studied viruses. For studies on cell‐associated virus, H9 cells productively infected with HIV were added to platelet suspensions and treated with the above regimen of AMT and UVA. The phototreated cells were then cocultivated with uninfected H9 cells for 4 weeks and supernatants were assayed by enzyme‐linked immunosorbent assay for HIV p24. No evidence of HIV replication was detectable for cells receiving as little as 2.4 J per cm2 of UVA irradiation in the presence of AMT. Further, it has been demonstrated that stably integrated sequences from the HIV proviral env gene can no longer be amplified by polymerase chain reaction after 1.2 J per cm2 of UVA (with 40 micrograms/mL AMT) exposure. CONCLUSION: These data suggest that AMT and UVA is an effective antiviral treatment for free and cell‐ associated HIV in platelet suspensions
Gene frequencies of the five major human platelet antigens in African American, white, and Korean populations
Authors: Kim H.O. , Jin Y. , Kickler T.S. , Blakemore K. , Kwon O.H. , Bray P.F.
Published: March 2003

BACKGROUND: The study of the immunogenetics of the human platelet antigens is important to the improvement of diagnosis and genetic counseling and to the development of screening programs for women at risk of having babies with neonatal alloimmune thrombocytopenia. Description of the immunogenetics of the human platelet antigens in some racial groups has been incomplete.

STUDY DESIGN AND METHODS: A reverse dot blot technique employing polymerase chain reaction‐ amplified genomic DNA was applied in genotyping the five major human platelet antigens in the following populations: 100 African American and 100 white women admitted to the obstetric unit at Johns Hopkins Hospital (Baltimore, MD) and 100 inpatients at Yonsei University (Seoul, Korea).

RESULTS: The gene frequencies of HPA‐2b (Koa) and HPA‐ 5b (Bra) in African Americans were twice those in whites (African Americans: 0.18 and 0.21, respectively; whites: 0.09 and 0.11, respectively). There is a very low gene frequency of the HPA‐1b (PIA2) allele in Koreans (0.005). No significant differences were found in the gene frequencies of the human platelet antigens in whites in this series and those in published European studies.

CONCLUSION: These studies indicate a higher potential risk for alloimmunization to HPA‐2 (Ko) and HPA‐5 (Br) antigens in African Americans than in whites. In addition, the low gene frequency of HPA‐1b (PIA2) in African Americans and Koreans suggests that alloimmunization to HPA‐1a (PIA1) would be very unusual in these populations. These data may provide the basis for planning neonatal alloimmune thrombocytopenia screening programs in certain ethnic populations.
Selective protection against IgG binding to red cells treated with phthalocyanines and red light for virus inactivation
Authors: Rywkin S. , Ben‐Hur E. , Reid M. E. , Oyen R. , Ralph H. , Horowitz B.
Published: March 2003

BACKGROUND: Irradiation with red light of red cells (RBCs) containing the photodynamically active phthalocyanine (Pc) dyes is being studied for inactivation of lipid‐enveloped viruses. One of the outstanding problems with this treatment is the binding of IgG to RBCs. The effects of oxygen and type I or type II quenchers on this IgG uptake were evaluated.

STUDY DESIGN AND METHODS: The Pc compounds used were aluminum phthalocyanine tetrasulfonate (AIPcS4), HOSiPcOSi(CH3)2(CH2)3N(CH3)2 (Pc 4); HOSiPcOSi(CH3)2(CH2)3N+(CH3)3I‐ (Pc 5); and SiPcOSi[(CH3)2(CH2)3N+(CH3)3](2)2I‐ (Pc 6). RBCs were analyzed by flow cytometry for the presence of IgG.

RESULTS: Irradiation with red light for 30 minutes of RBCs containing either 2 microM Pc 4, 2 microM Pc 5, 2 microM Pc 6, or 6.5 microM AIPcS4 resulted in an uptake of IgG. These conditions completely inactivated the lipid‐enveloped vesicular stomatitis virus (VSV) (> 5 log10 kill). IgG uptake was reduced when oxygen was depleted. The addition of reduced glutathione (GSH) or mercaptoethanol prevented the binding of IgG with RBCs treated with AIPcS4, Pc 4, Pc 5, and Pc 6. Specific binding of IgG2 but not of C3d was observed upon irradiation of RBCs with Pc 5 and Pc 6 in the absence of GSH. No gross changes were observed in RBC antigen strength after irradiation with the dyes in the presence of GSH. Inactivation of VSV by Pc plus light was not affected by GSH.

CONCLUSION: Sulfhydryl compounds are useful in preventing IgG binding to RBCs following Pc photosensitization. Since virus inactivation proceeds at the same rate in the presence and the absence of sulfhydryl compounds, their addition to treated RBCs should allow crossmatching for transfusion after treatment. The binding of IgG depends to a large extent on the generation of reactive oxygen species.
Analysis of the expression of NB1 antigen using two monoclonal antibodies
Authors: Stroncek D. F. , Shankar R. A. , Norën P. A. , Herr G. P. , Clement L. T.
Published: March 2003

BACKGROUND: Neutrophil‐specific antigen NB1 is expressed on neutrophil subpopulations in 97 percent of healthy individuals and is located on 56‐ to 64‐kDa glycoprotein. While the molecule carrying NB1 has been identified, the nature of the NB1 epitope has not been well characterized.

STUDY DESIGN and METHODS: Two monoclonal antibodies (MoAbs), 1B5 and the recently produced 7D8, and four alloantibodies, all specific for NB1, were used to investigate the expression of NB1 on neutrophils from several donors.

RESULTS: MoAb 7D8 was shown to be specific for NB1. It reacted with NB1‐positive neutrophils from 52 donors in the granulocyte immunofluorescence assay and did not react with NB1‐negative neutrophils from 8 donors. MoAb 7D8 immunoblotted a 56‐ to 64‐kDa molecule on neutrophils from eight NB1‐positive donors and did not react with this molecule on NB1‐negative neutrophils from two donors. When 7D8 was tested in the monoclonal antibody immobilization of granulocyte antigens assay, it reacted with two NB1 alloantibodies, but not with NA1 or NA2 alloantibodies. To determine if MoAbs 7D8 and 1B5 recognized the same epitope, both were tested against the same NB1‐positive neutrophils and the cells were analyzed by two‐ color flow cytometry. Both antibodies bound independently to neutrophils, which indicated that the antibodies recognized different epitopes. When similar studies were performed with MoAb 7D8 and three NB1 alloantibodies, 7D8 partially inhibited the binding of two of the alloantibodies. The size of the NB1‐positive subpopulation was analyzed in 25 people using flow cytometry with both MoAbs and three alloantibodies. The subpopulation of antigen‐positive cells was similar in all donors when 7D8 and the three NB1 alloantibodies were tested; however, the subpopulation recognized by MoAb 1B5 was smaller in two of the donors. Neutrophils from one of these people were analyzed by immunoblotting, and no differences were detected in the molecule carrying NB1 in those neutrophils and that molecule in control neutrophils.

CONCLUSION: NB1 specificity is made up of at least two separate epitopes. The expression of NB1 varied among antigen‐positive individuals. While NB1 is expressed by a 56‐ to 64‐kDa glycoprotein, the structure of this protein on antigen‐negative cells has not been determined.
A prospective, randomized study of the use of platelet concentrates irradiated with ultraviolet‐B light in patients with hematologic malignancy
Authors: Blundell E.L. , Pamphilon D.H. , Fraser I.D. , Menitove J.E. , Greenwalt T.J. , Snyder E.L. , Repucci A.J. , Hedberg S.L. , Anderson J.K. , Buchholz D.H. , Kagen L.R. , Aster R.H.
Published: March 2003

Background: Irradiation of platelet concentrates (PCs) with ultraviolet‐ B (UVB) light inactivates the contaminating white cells and might be an alternative to filtration for the prevention of alloimmunization to HLA antigens and subsequent refractoriness to further platelet transfusions in multiply transfused patients with bone marrow failure.

Study Design and Methods: Patients with hematologic malignancy, mainly acute myeloid leukemia, were prospectively assigned in a random manner to receive either UVB‐irradiated or control, nonirradiated PCs. All patients were given red cells that were white cell reduced by filtration. Transfusion efficacy and alloimmunization were assessed by means of corrected count increments, requirement for red cells and PCs, and measurement of lymphocyte‐reactive antibodies.

Results: UVB‐irradiated PCs had a clinical efficacy similar to controls as judged by corrected count increments at 1 to 6 and 12 to 24 hours and by the median requirement for red cell and platelet transfusions. Alloimmunization determined by measurements of lymphocyte‐reactive antibodies using both conventional and antiglobulin‐augmented lymphocytotoxicity techniques was not abolished in recipients of UVB‐irradiated PCs (4/30, 13%) but was less than that in controls (5/20, 25%; p = NS). The mean number of platelet transfusion episodes prior to the occurrence of alloimmunization was greater in the control group (27 vs. 10; p = 0.017).

Conclusion: In this trial, UVB irradiation did not diminish the clinical efficacy of platelet transfusions. There was a small but nonsignificant reduction alloimmunization, but no difference in refractoriness of the two groups was observed. Larger prospective randomized studies are required to confirm these findings and to compare UVB irradiation with white cell reduction.
Polymorphisms at the ABO locus in subgroup A individuals
Authors: Olsson M.L. , Chester M.A.
Published: March 2003

Background: The common ABO allele sequences are known, but little or no genetic information is available on the rare but important A subgroups.

Study Design and Methods: Blood group ABO polymorphism was analyzed in genomic DNA from 45 rare subgroup A individuals by sequence‐specific primer polymerase chain reaction and amplified fragment length polymorphism investigating exons VI and VII in the ABO genes. These methods are used to detect specific mutations only, and not all changes that might be present can be detected. ABO genotypes discriminating six alleles (A1, A2, B, O1, O1var, and O2) were determined.

Results: The C→T substitution at nucleotide position 467 (C467T) is not restricted to A2 and cis‐AB individuals, but was found also in some A subgroups. Detection of the functionally more relevant C1060‐single‐point deletion in A2 was accomplished by a novel sequence‐specific primer polymerase chain reaction approach. A 100‐percent correlation between the C467T and the C1060‐mutations was found. Fifteen of 17 samples showing the T646A mutation (described earlier in one case of Ax) showed a positive correlation with the C771T mutation in a frequently occurring O1var allele. The two exceptions were defined serologically as Ax.

Conclusion: Indications have been found of an evolutionary relationship between A1 alleles and Ael and A3 subgroups as well as between A2 alleles and Aend and Aweak subgroups. Genetic heterogeneity within the Ax and Aint subgroups was also seen.
Optimal conditions for white cell reduction in red cells by filtration at the patient's bedside
Authors: Sirchia G. , Rebulla P. , Sabbioneda L. , Garcea F. , Greppi N.
Published: March 2003

Background: A quality control program of white cell (WBC) reduction in red cells at the bedside was implemented, based on postfiltration counting in a Nageotte chamber of the residual WBCs from samples taken from a segment of the transfusion set, after 1‐in‐10 sample dilution with Turks's solution. During a 1‐year quality control program, 5.1 percent of counted units had apparent filtration failures, that is, WBC counts exceeding 5 × 106 per unit. The cause(s) for these apparent failures were investigated.

Study Design and Methods: In Study 1, residual WBCs from 150 buffy coat‐free red cells filtered through one type of filter at 4°C, 20 to 24°C, or 27°C in 5 to 10 minutes, 50 to 100 minutes, or 100 to 200 minutes were counted as described above. In Study 2, residual WBCs in samples collected from segments of the transfusion set and from the postfiltration bags were counted in parallel by a new, more sensitive counting method. In this method, 5 mL of filtered red cells was diluted with 20 mL of 3‐percent paraformaldehyde and centrifuged, the pellet was resuspended to 500 μL with a lysis solution, and the WBCs were counted in a Nageotte chamber. In Study 3, residual WBCs were counted by the 3‐percent paraformaldehyde method in samples from postfiltration bags of 1‐ to 2‐ day‐old buffy coat‐rich red cell units filtered through a second type of filter. Filtration was started within 30 minutes of the removal of the unit from the refrigerator, ambient temperature was 20 to 24°C, and the median filtration time was 90 minutes per unit.

results: Study 1: Median WBC counts per unit increased progressively from 51,000 at 4°C to 934,000 at 27°C, with intermediate values at 20 to 24°C. In no unit did the WBC count exceed 5 × 106 if filtration at 20 to 24°C was completed within 100 minutes, while counts in excess of 50 × 106 were found at 20 to 24°C and at 27°C with filtration times of 100 to 200 minutes, and 50 to 100 minutes, respectively. Study 2: The relation between segment and postfiltration bag WBC counts obtained by the 3‐ percent paraformaldehyde method was poor, with the latter being almost always lower than the former. Study 3: None of the 120 units filtered through the second type of filter at 20 to 24°C in 50 to 100 minutes contained more than 3.2 × 106 WBCs; the median value was 147,000 WBCs per unit.

Conclusion: On the basis of the results with the 3‐percent paraformaldehyde method, which showed the unreliability of segment counts, a new policy was adopted for quality control of bedside WBC reduction, based on controlling the time of and temperature at transfusion. Bedside WBC reduction in 1‐ to 2‐day‐old red cells performed with the second type of filter at 20 to 24°C in less than 100 minutes per unit allowed the preparation of units that meet the standard of fewer than 5 × 106 WBCs in all tested cases. Bedside WBC reduction with the second type of filter and under the controlled conditions reported seems effective.
HLA class I‐eluted platelets as an alternative to HLA‐matched platelets
Authors: Novotny V. M. , Huizinga T. W. , Van Doorn R. , Briet E. , Brand A.
Published: March 2003

BACKGROUND: Alloimmunized refractory thrombocytopenic patients often require HLA‐matched platelet transfusions. As the HLA system is very polymorphic, sufficient HLA‐matched donors are not available for every patient.

STUDY DESIGN AND METHODS: In vitro elution techniques with citric acid incubation of platelets at pH 3.0 showed that platelets lose expression of HLA, whereas platelet‐specific glycoproteins are preserved. This technique was modified for clinical use. Random‐donor platelet concentrates were incubated with citric acid, subsequently washed, and transfused to two patients.

RESULTS: Platelet‐specific glycoproteins were unaffected, and HLA expression decreased generally to below 25 percent of the initial expression. One alloimmunized patient who was without compatible donors because of a rare HLA type underwent repeated transfusions with acid‐treated platelets. In contrast to the results with random‐donor platelet transfusions, posttransfusion increments up to 47 × 10(9) per L were obtained with acid‐treated platelets, and profuse gastrointestinal bleeding was stopped, while multiple skin hemorrhages were resolved. No side effects were observed. A second patient developed a severe transfusion reaction without platelet increment after one transfusion with acid‐treated platelets expressing 30 percent of the original HLA antigens. Further transfusions were not given.

CONCLUSION:Standardization of the acid elution technique and validation of the technique in patients is necessary. The results suggest, however, that HLA‐eluted platelets prepared under specified conditions may gain a place in platelet transfusion therapy.
Safety and efficacy of hepatitis C virus antibody screening of blood donors with two sequential screening assays
Authors: Allaina J.P. , Kitchens A. , Aloysiuis S. , Reeves I. , Petrik J. , Barbara J. A. J. , Williamson L.M.
Published: March 2003

BACKGROUND: Reactive samples in hepatitis C virus (HCV) antibody screening of blood donors are currently referred for a confirmatory assay. This scheme is not optimally efficient and is expensive because of the lack of specificity and cost of confirmatory tests, as well as the need to discard false‐positive donations. As in some human immunodeficiency virus antibody‐confirmatory schemes, the safety and efficacy of confirming anti‐HCV with two sequential screening assays were evaluated.

STUDY DESIGN AND METHODS: Three combinations of two anti‐HCV screening assays were used to test 75,874 blood donors. Results were compared with the routine testing scheme and HCV RNA detection in any enzyme immunoassay‐repeatably reactive samples.

RESULTS: The use of an alternative screening assay for repeat testing decreased the proportion of enzyme immunoassay‐positive donors from 0.28 to 0.05 percent. All samples that were “confirmed” as positive by the standard combination of immunoassays and all HCV RNA‐positive samples were detected by the sequential screening assays. No samples that had discordant results on primary and secondary screening assays were confirmed by recombinant immunoblot assay or were found to contain detectable HCV RNA.

CONCLUSION: The combination of screening assays for anti‐HCV confirmation was as safe as, cheaper than, and nearly as efficient as the standard testing scheme.
Flow cytometric quantitation of serum anti‐D in pregnancy
Authors: Christensson M. , Bremme K. , Shanwell A. , Westgren M. , Christensson B.
Published: March 2003

BACKGROUND: The major cause of fetal hemolytic disease is maternal immunization to D in D‐incompatible pregnancies. To prevent complications, D‐incompatible pregnancies are monitored for the level of maternal anti‐D. At present, the monitoring of anti‐D levels is performed by the indirect antiglobulin test complemented by quantitation by the technique used in an automated antibody detection and quantitation instrument. STUDY DESIGN AND METHODS: Flow cytometry was used to quantitatively determine the level of anti‐D in serum and to analyze the IgG subclass distribution and the presence of IgM anti‐D in these samples. The results were compared to the indirect antiglobulin test titer and to the results obtained by the technique used in an automated antibody detection and quantitation instrument. RESULTS: Flow cytometry allowed sensitive and accurate determinations of anti‐D levels with low interassay and intra‐assay variability, both for serum samples and standard curves. CONCLUSION: Flow cytometry is a simple, rapid, and reliable method for determining the serum levels of D antibodies and their Ig subclass distribution. It is therefore well suited for the monitoring of women during D‐incompatible pregnancies.
A randomized, double‐blind comparison of donor tolerance of 400 mL, 200 mL, and sham red cell donation
Authors: Smith K. J. , James D. S. , Hunt W. C. , McDonough W. , Quintana R.
Published: March 2003

BACKGROUND: Volume replacement could allow the safe collection of twice the normal amount of red cells in a standard donation. Studies in small numbers of donors have shown that a temporary decrease in red cell mass is well tolerated when donors give twice the usual amount (170–225 mL) of red cells in a standard 405‐ to 495‐mL donation. Sham‐donation control groups have not been included in previous studies of increased red cell donation, and perceptions of donation effects could have been biased. STUDY DESIGN AND METHODS: In the study reported here, 17 male and 13 female volunteers were randomly assigned to make a sham donation, 1‐unit donation, or 2‐unit donation on an automated blood cell separator. Donor tolerance was assessed by ambulatory heart rate monitoring and by a poststudy interview. Hemoglobin, hematocrit, ferritin, serum iron, total iron‐binding capacity, red cell 2,3 DPG, and serum erythropoietin were measured before and after donation for comparison of the erythropoietic responses in the three study groups. RESULTS: Red cells collected totaled 206 +/− 10 mL in the 1‐unit group and 414 +/− 21 mL in the 2‐unit group. Changes in heart rate, systolic blood pressure, and diastolic blood pressure with donation and changes in heart rate recorded by ambulatory monitoring did not differ for the experimental groups. Postdonation changes from baseline values were evaluated on Days 2, 7, and 14. Changes in hemoglobin were significantly different between groups (p < 0.017) in all postdonation tests. There were differences between groups in erythropoietin response, red cell 2,3 DPG, ferritin levels, and hemoglobin synthesis. Hemoglobin synthesis and mean changes in 2,3 DPG, erythropoietin, ferritin, and postdonation hemoglobin were greater in the 2‐unit group than in the 1‐unit group. CONCLUSION: Donor tolerance of red cell donations of 414 +/− 21 mL, a volume of red cells twice that in a standard 450‐mL blood donation, does not differ from donor tolerance of standard or sham donations. Physiologic adjustments and the hematopoietic response to reduced red cell mass were greater in the 2‐ unit group, but the donation of 1 unit or 2 units did not cause detectable symptoms of reduced oxygen‐carrying capacity.
A highly sensitive and specific chemiluminescent enzyme‐linked immunosorbent assay for diagnosis of active Trypanosoma cruzi infection
Authors: Almeida Igor C. , Covas Dimas T. , Soussumi Lea M.T. , Travassos Luiz R.
Published: March 2003

BACKGROUND: Chagas' disease is transmitted to man either by the bite of insects harboring Trypanosoma cruzi or by the transfusion of blood from infected donors. The conventional serologic testing as presently used in blood banks in South America is unsatisfactory, because of a high number of inconclusive and false‐positive results. Other methods such as polymerase chain reaction and enzyme‐linked immunosorbent assay (ELISA) with recombinant antigens have been proposed, but inherent difficulties have so far precluded their adoption in the large‐scale screening required by blood banks. STUDY DESIGN AND METHODS: A highly sensitive and specific chemiluminescent ELISA using a purified trypomastigote glycoconjugate antigen and a complex epimastigote antigen was devised for the diagnosis of active T. cruzi infection. RESULTS: Chemiluminescent ELISA was 100‐percent sensitive in the diagnosis of 100 cases of confirmed Chagas' disease. Inconclusive results and false‐positive reactions were eliminated in a panel of 115 sera. The specificity of the chemiluminescent ELISA was 100 percent with a purified trypomastigote glycoconjugate antigen and 99.7 percent with a complex epimastigote antigen when applied to 1000 normal human sera and 288 heterologous sera from patients with other infections, including leishmaniasis, and vaccinated individuals. CONCLUSION: The chemiluminescent ELISAs provide a test that is highly sensitive (purified trypomastigote glycoconjugate and complex epimastigote antigens) and specific (purified trypomastigote glycoconjugate antigen) for Chagas' disease diagnosis. It can be used in blood bank screening and to monitor the treatment of patients undergoing chemotherapy.