bluuebunny/crossref_metadata_2025 huggingface dataset:
Validation of use of the Nageotte hemocytometer to count low levels of white cells in white cell‐reduced platelet components
Authors: Moroff G. , Eich J. , Dabay M.
Published: March 2003

BACKGROUND: Determination of the white cell (WBC) count in WBC‐reduced platelet components requires methods that have a detection limit in the range of approximately 5.0 × 10(2) to 5.0 × 10(4) per mL.

STUDY DESIGN AND METHODS: With a 50‐microL Nageotte hemocytometer and bright‐field microscopy (200x magnification), studies were conducted to develop and validate a method that could be used routinely with filtered and apheresis‐harvested platelets. A 1‐in‐5 dilution of sample with a commercially available blood‐diluting fluid was used because, with a lower (1‐in‐2) dilution, the observed number of WBCs was substantially less than the number expected at relatively high platelet counts (> 1.9 × 10(9)/mL).

RESULTS: The observed and expected WBC counts in WBC‐ reduced platelet samples correlated well at levels between approximately 5 and 1100 WBCs per counting area (5.0 × 10(2)‐1.1 × 10(5)/mL). At levels of more than 300 to 400 WBCs per counting area, accurate counts were obtained when 10 of the 40 rectangles were counted.

CONCLUSION: These studies provide data to confirm that the 50‐ microL Nageotte hemocytometer can be used to accurately count low levels of WBCs in platelet components.
What can we learn about the need for transfusion from patients who refuse blood? The experience with Jehovah's Witnesses
Authors: Viele M.K. , Weiskopf R.B.
Published: March 2003

BACKGROUND: A transfusion threshold of 7 g per dL (70 g/L) of hemoglobin has been proposed for patients, although scant human data are available to support this recommendation.

STUDY DESIGN AND METHODS: The medical community's experience with Jehovah's Witnesses was examined, in order to assess the lowest tolerable hemoglobin concentration and the lower transfusion threshold of 7 g per dL (70 g/L) of hemoglobin. A MEDLINE search was conducted to capture medical and surgical reports involving Jehovah's Witnesses from 1970 through early 1993.

RESULTS: Sixty‐one reports of untransfused Jehovah's Witnesses with hemoglobin concentrations < or = 8 g per dL (80 g/L) or hematocrits < or = 24 percent (0.24) were identified. Of 50 reported deaths, 23, as stated in the original reports, were primarily due to anemia. Except for three patients who died after cardiac surgery, all patients whose deaths were attributed to anemia died with hemoglobin concentrations < or = 5 g per dL (50 g/L). Twenty‐five survivors were reported with hemoglobin < or = 5 g per dL (50 g/L).

CONCLUSION: These data have significant limitations but suggest that survival, without transfusion, is possible at low hemoglobin concentrations, while mortality with an unknown incidence is encountered at hemoglobin concentrations below 5 g per dL (50 g/L).
Blood use in patients undergoing repeat coronary artery bypass graft procedures: multivariate analysis
Authors: Bracey A.W. , Radovancevic R. , Radovancevic B. , McAllister H.A. , Vaughn W.K. , Cooley D.A.
Published: March 2003

BACKGROUND: The prevailing clinical opinion is that patients undergoing repeat coronary artery bypass graft (CABG) operation require more blood transfusions than do patients undergoing primary CABG operation. To determine the extent of this increased demand and the variables responsible for it, the cases of 196 patients who had undergone primary procedures and 65 patients who had had repeat procedures at the same institution were reviewed.

STUDY DESIGN AND METHODS: To analyze the differences in transfusion requirements for these two groups, the following data were obtained: number of transfusions given between the time of surgery and the time of hospital discharge; preoperative hemoglobin (Hb), hematocrit (Hct), prothrombin time, and platelet count; Hb and Hct at hospital discharge; time the patient was on cardiopulmonary bypass; number and type of grafts; estimates of intraoperative blood loss; and chest‐tube blood shed during the first 48 hours after surgery.

RESULTS: The groups were comparable with respect to age, body weight, preoperative Hb and Hct, number of grafts, and aspirin exposure. Patients in the repeat group had 35‐percent greater blood loss and required 75‐percent more blood components than did the patients undergoing primary procedures. The mean number of blood components transfused per patient was as follows: red cells, 3.8 +/− 0.5 units in repeat patients and 2.2 +/− 0.2 units in primary patients (p = 0.002); platelets, 2.9 +/− 0.9 vs. 1.1 +/− 0.2 (p = 0.043); fresh‐frozen plasma, 1.6 +/− 0.4 vs. 0.8 +/− 0.1 (p = 0.044). Analysis of variables by regression method for repeat patients showed a predictive effect of blood loss (p < 0.0001), prolonged time on cardiopulmonary bypass (p < 0.0001), preoperative Hb (p = 0.0003), and aspirin exposure (p = 0.0094) on red cell transfusion rate in repeat patients (R‐square = 0.7778, Prob > f = 0.0001).

CONCLUSION: Repeat CABG patients have higher transfusion rates. These findings may be attributed to the increased microvascular bleeding, prolonged time on cardiopulmonary bypass, lower preoperative Hb, and the use of preoperative antiplatelet medications.
Transfusion‐associated graft‐versus‐host disease: risk due to homozygous HLA haplotypes
Authors: Wagner F. F. , Flegel W. A.
Published: March 2003

BACKGROUND: Transfusion‐associated graft‐versus‐host disease (TA‐GVHD) may occur in transfusions of blood from HLA‐homozygous persons to HLA‐ heterozygous persons who share a haplotype. STUDY DESIGN AND METHODS: Two mathematical models were developed to calculate the upper and lower limit of the associated risks in various populations using a combination of serology‐ and DNA sequence‐based HLA haplotype frequencies. RESULTS: For nondirected transfusion, the range of the estimated risk in United States whites is 1 of 17,700 to 39,000 (1/6,900‐48,500 in Germans; 1/1,600‐7,900 in Japanese). The risk in directed donation between parents and children is increased at least 21‐ fold for US whites, 18‐fold for Germans, and 11‐fold for Japanese. CONCLUSION: For nondirected transfusions, the estimates of TA‐GVHD risk derived in this model are lower than estimates of previously published models, are in better agreement with the clinical experience, and explain in part the observed discrepancy between TA‐GVHD incidence in the United States and that in Japan. Most notably for US whites, the relative increase in risk in directed transfusion is larger than previously thought.
Posttransfusion hepatitis after induction chemotherapy in acute nonlymphoblastic leukemia: implications for long‐term management and outcome
Authors: López‐Jimenez J. , Cancelas J. A. , García‐Laran̄a J. , Sastre J. L. , Cerveró C. , Zamora C. , Megido M. , Hernández‐Jodra M. , Lasa E. , Pérez‐Oteyza J. , Ramos P. , Odriozola J. , Navarro J. L.
Published: March 2003

BACKGROUND: The purpose of this study was to evaluate 1) the incidence of hepatitis and its influence on the clinical management of and outcome in acute nonlymphoblastic leukemia (ANLL) patients in first complete remission and 2) the impact of routine hepatitis C virus screening on the incidence of hepatitis in these patients. STUDY DESIGN AND METHODS: Clinical and blood bank charts were reviewed for 65 consecutive ANLL patients between 1985 and 1993 who achieved complete remission after a course of daunomycin and cytarabine (cytarabine: 200 mg/m2/day × 7 days in continuous infusion; daunomycin: 60 mg/m2/day for the first 3 days of the 7, as a bolus). RESULTS: Only 43 percent of patients who developed hepatitis completed the scheduled therapy. Hepatitis did not decrease the probability of relapse (66 +/− 9% vs. 66 +/− 11%), but patients with changes in planned treatment, due to hepatitis or other causes, tended to have a higher relapse rate than patients without changes in consolidation therapy (56.5% vs. 40.4%; p = 0.10). This did not result in a decrease in disease‐free survival, however, because of the higher number of treatment‐related deaths in the patients without hepatitis (who completed the therapy). Over the period from 1985 through 1989, the 6‐month actuarial probability of developing hepatitis was 42 percent. However, since 1989, when hepatitis C screening of blood donors was implemented, the incidence was reduced to 12.5 percent (p < 0.05), in spite of greater transfusion support (172 +/− 46 vs. 89 +/− 53, p < 0.01). No new cases of hepatitis were observed after the introduction of second‐generation hepatitis C virus assays. CONCLUSION: Hepatitis precludes the use of consolidation therapy in about half of ANLL patients, resulting, in the experience described here, in a trend toward a higher rate of relapse. Hepatitis C virus screening of blood components reduces the incidence of hepatitis in ANLL patients.
Albumin batches and B19 parvovirus DNA
Authors: Lefrère J. J. , Mariotti M. , de la Croix I. , Lerable J. , Thauvin M. , Burnouf T. , Folléa G.
Published: March 2003

BACKGROUND: B19 parvovirus (B19) may be transmitted iatrogenically by blood, and its prevalence in blood donations is estimated at 1 in 3,300 to 1 in 50,000. As a large number of blood donations make up the plasma pools used to produce plasma derivatives, even a virus as rare as B19 in a population of blood donors may result in the frequent contamination of plasma batches. The percentage of albumin batches containing B19 DNA has never been determined.

STUDY DESIGN AND METHODS: The presence of B19 DNA was investigated by a polymerase chain reaction assay (with a primer pair in the VP1 region) in a total of 12 and 17 batches of 4‐ and 20‐percent albumin, respectively, from two different manufacturers.

RESULTS: No B19 DNA was detected in the batches tested.

CONCLUSION: The current fractionation process used to obtain these albumin preparations is seen to allow the efficient degradation and/or elimination of B19.
Reliability of the third‐generation recombinant immunoblot assay for hepatitis C virus
Authors: Damen M. , Zaaijer H.L. , Cuypers H.T. , Vrielink H. , van der Poel C.L. , Reesink H.W. , Lelie P.N.
Published: March 2003

BACKGROUND: In a confirmatory laboratory, the second‐generation recombinant immunoblot assay (RIBA‐2) was replaced by the third‐ generation RIBA (RIBA‐3) in March 1993. The aim of this validation study was to compare the sensitivity and specificity of RIBA‐2 and RIBA‐ 3 in a routine setting, by using a validated hepatitis C virus (HCV) RNA polymerase chain reaction to establish plasma viremia.

STUDY DESIGN AND METHODS: RIBA‐2 testing was performed (March 1991‐March 1993) in 593 HCV RNA‐positive and 1498 HCV RNA‐negative subjects. RIBA‐3 testing was performed (March 1993‐May 1994) in 220 HCV RNA‐positive and 530 HCV RNA‐negative subjects. All samples reacted for anti‐HCV in enzyme‐ linked immunosorbent assay.

RESULTS: In HCV RNA‐positive individuals, the sensitivity of RIBA‐3 was significantly higher than that of RIBA‐2 (99.5% vs. 93.3%, p = 0.0005). This was not caused by inclusion of the NS5 antigen, but by a higher sensitivity of the antigens c33 and c100 (RIBA‐2: 94.3% and 62.6%; RIBA‐3: 99.5% and 88.6%). Replacement of the c22 and c100 recombinant proteins by synthetic peptides significantly reduced nonspecific reactivity against these antigens (p < 0.0001). Unfortunately, increased nonspecific reactivity against the modified c33 antigen and the new NS5 antigen canceled out this effect. Two‐band reactivity occurred more often in nonviremic persons than in viremic persons (32.7% vs. 8.2%, p < 0.0001). Risk factors for HCV infection were less frequently observed in 11 blood donors with two‐band reactivity than in 6 blood donors with other positive RIBA‐3 patterns (18% vs. 83%, p = 0.03).

CONCLUSION: The higher sensitivity of RIBA‐3 significantly reduced the number of indeterminate test results in HCV RNA‐positive persons. Confirmatory laboratories must be aware of the frequent occurrence of nonspecific, isolated reactivity and even nonspecific, two‐band reactivity in anti‐HCV enzyme‐linked immunosorbent assay‐reactive blood donors.
Time‐dependent, spontaneous release of white cell‐ and platelet‐derived bioactive substances from stored human blood
Authors: Nielsen H. J. , Reimert C. M. , Pedersen A. N. , Brünner N. , Edvardsen L. , Dybkjær E. , Kehlet H. , Skov P. S.
Published: March 2003

BACKGROUND: The mechanisms of the detrimental effects of perioperative allogeneic blood transfusion are still unclear. Previous studies have suggested a higher incidence of adverse effects after the use of blood stored for prolonged time. Therefore, a possible time‐dependent release of various white cell‐ and platelet‐derived bioactive substances in stored human red cell suspensions was studied. STUDY DESIGN AND METHODS: Whole blood (6 units), plasma‐reduced whole blood (6 units), and saline‐adenine‐glucose‐mannitol blood (6 units) from 18 unpaid, normal blood donors were stored under standard blood bank conditions at 4 degrees C for 35 days. After refrigeration, samples were collected from all blood bags on Days 0, 2, 5, 9, 14, 21, 28, and 35 of storage. Extracellular concentrations of eosinophil cationic protein, eosinophil protein X, plasminogen activator inhibitor 1, myeloperoxidase, and interleukin 6 were analyzed by enzyme‐linked immunosorbent assay and radioimmunoassay. The total intracellular and donor plasma levels of these substances also were analyzed at the time of blood donation. RESULTS: Eosinophil cationic protein, eosinophil protein X, and myeloperoxidase increased 10‐ to 25‐fold (p < 0.05) in a time‐dependent manner in whole blood, plasma‐reduced whole blood, and saline‐adenine‐ glucose‐mannitol blood during storage for 35 days. Plasminogen activator inhibitor 1 increased threefold to sixfold (p < 0.05) in whole blood and plasma‐reduced whole blood, but not in saline‐adenine‐ glucose‐mannitol blood. Interleukin 6 was not detected in either plasma or samples obtained from the blood bags. CONCLUSION: Stored whole blood, plasma‐reduced whole blood, and saline‐adenine‐glucose‐mannitol blood may release white cell‐ and platelet‐derived bioactive substances in a time‐dependent manner, which may be related to the detrimental effects of perioperative blood transfusions. Therefore, prestorage white cell reduction should be considered for further improvement of red cell suspensions.
Risk of exposure to Chagas' disease among seroreactive Brazilian blood donors
Authors: Salles N. A. , Sabino E. C. , Cliquet M. G. , Eluf‐Neto J. , Mayer A. , Almeida‐Neto C. , Mendonça M. C. , Dorliach‐Llacer P. , Chamone D. F. , Saéz‐Alquézar A.
Published: March 2003

BACKGROUND: Screening of blood donors for Chagas' disease by using currently available serologic tests is complicated by the lack of adequate sensitivity, discordant results between tests, and the absence of a gold standard. STUDY DESIGN AND METHODS: The study was designed to evaluate the serologic tests by using epidemiologic data relating to the risk of exposure to Trypanosoma cruzi in the urban centers of Brazil. The serologic results obtained from screening 411,617 voluntary blood donations in Sao Paulo during 1993 and 1994 were reviewed, as well as follow‐up results on 1,267 donors who initially were repeatably reactive in at least one of three screening tests. Epidemiologic data were obtained from 321 individuals who on follow‐up remained reactive in at least one test and who returned for medical counseling. Controls included 119 screen‐negative blood donors and 45 blood donors who were repeatably reactive in at least one screening test but were negative on follow‐up. RESULTS: Of the individuals who reacted in three screening tests, 94.6 percent remained reactive on follow‐up. Of the individuals who were repeatably reactive in only one screening test, 70.8 percent were negative in all three tests on follow‐up. Most individuals who reacted in two or three tests on follow‐up had epidemiologic evidence of a risk of exposure to Chagas' disease. A significant proportion (29.1%) of those who were reactive in only one test on follow‐up had epidemiologic evidence of exposure to the Chagas' disease vector as compared to 14.6 percent of controls (p = 0.007). This suggests that some of these individuals truly were infected. CONCLUSION: No single test for Chagas' disease is sufficiently sensitive to prevent transfusion transmission of the disease in the urban centers of Brazil.
Effect of blood donation on the establishment of normal ranges of lymphocyte subsets
Authors: Bryant J. A. , Wylie B. R. , Yuan F. F. , Ribeiro A. , Thomson A. R. , Cooley M. A. , Fletcher A.
Published: March 2003

BACKGROUND: Absolute counts of CD4+ T‐lymphocytes are used in the management of patients with human immunodeficiency virus infection. Low absolute counts of CD3+CD4+ cells have also been observed in healthy people–a phenomenon called idiopathic CD4 lymphocytopenia. It is common practice for normal ranges for lymphocyte subsets to be derived from samples taken from blood donors. STUDY DESIGN AND METHODS: A sample of EDTA blood was taken through the donation line tubing, after donation from 565 blood donors in Sydney, Australia, who were selected from a range of age groups. An additional 12 donors provided a predonation sample as well as a postdonation sample. Hematologic assays were performed on two analyzers. Samples were stained for CD3, CD4, CD8, CD19, and CD56 and analyzed on a flow cytometer. RESULTS: Three donors were found to have absolute CD3+CD4+ counts < 300 cells per microL. The percentage of CD3+CD4+ cells was found to increase with age. Both the percentage and the absolute count of CD3+CD8+ cells decreased with age, which resulted in an increased CD4:CD8 ratio with age. Men had consistently higher absolute counts of CD3‐CD56+ cells than women. The 12 additional donors all had greater percentages of CD3+CD4+ cells and lower absolute counts for CD3+, CD3+CD4+, CD3+CD8+, CD19+ and CD3‐CD56+ cells after donation than they had before donation (p < 0.001). CONCLUSION: It is not satisfactory to base normal ranges for lymphocyte subsets on donor blood, from which the blood sample has been obtained after donation.
Unexpected citrate toxicity and severe hypocalcemia during apheresis
Authors: Uhl L. , Maillet S. , King S. , Kruskall M. S.
Published: March 2003

BACKGROUND: The citrate anticoagulant used during apheresis procedures is considered a safe medication because it is rapidly metabolized by the donor. However, acute, life‐threatening hypocalcemia is possible if the infusion rate of citrate is increased. CASE REPORT: A 54‐year‐old woman with metastatic breast cancer, but otherwise in good health, had just begun a fifth collection of hematopoietic peripheral blood progenitor cells by leukapheresis. The instrument's self‐loading apheresis kit was primed uneventfully. Seven minutes into the procedure, the patient developed signs and symptoms suggesting severe hypocalcemia, including muscle spasms, chest pain, and hypotension. The citrate bag was discovered to have emptied, and a section of the anticoagulant tubing was protruding outside of the rotary pump. The patient's ionized calcium level was 0.64 mmol per L (normal range, 1.18‐ 1.38 mmol/L). In subsequent experiments where the anticoagulant tubing was either improperly loaded at the outset or partially pulled out of the rotary pump, no instrument alarms sounded. CONCLUSION: Citrate toxicity and life‐threatening hypocalcemia can occur if the anticoagulant line of an apheresis instrument is not properly housed in its rotary pump or becomes disengaged during the procedure. Instrument manufacturers are encouraged to consider designs that allow the direct measurement of the volume of citrate delivered. In the interim, periodic visual and tactile confirmation of tubing placement during apheresis procedures is prudent.
Rejuvenation of irradiated AS‐1 red cells
Authors: Samuel L.H. , Anderson G. , Mintz P.D.
Published: March 2003

BACKGROUND: Gamma irradiation of blood components is used to prevent transfusion‐associated graft‐versus‐host disease. The demand for irradiated blood components is increasing because of the increase in directed donation by family members. Irradiated units currently have a recommended maximum storage life of 28 days. Since in vivo recovery is related to red cell ATP levels, rejuvenation of stored irradiated units using a pyruvate‐inosine phosphate‐adenine additive was explored.

STUDY DESIGN AND METHODS: Units of AS‐1 red cells from 16 volunteer donors were divided into two equal volumes and one split unit from each was irradiated with 25 Gy. Ten units were irradiated on Day 5, 6, or 7 of 4°C storage and 6 units were irradiated on Day 1 of 4°C storage. All units were rejuvenated for 1 hour at 37°C using a pyruvate‐inosine‐phosphate‐adenine additive on Day 42 of 4°C storage. Units were assayed for ATP, 2, 3 DPG and supernatant sodium, potassium, and glucose.

RESULTS: ATP and 2, 3 DPG levels were restored equally well in irradiated and non‐irradiated units. The previously reported irradiation‐induced red cell potassium‐sodium shift was demonstrated. Supernatant potassium and sodium levels did not reverse 1 hour after rejuvenation was completed. There was no significant difference in results between units irradiated on Day 1 or Day 5, 6, or 7.

CONCLUSION: Red cell ATP and 2, 3 DPG levels were restored in irradiated AS‐1 units stored at 4°C for 42 days using a pyruvate‐inosine‐phosphate‐adenine rejuvenation additive.
Rapid identification of bacterially contaminated platelets using reagent strips: glucose and pH analysis as markers of bacterial metabolism
Authors: Burstain J.M. , Brecher M.E. , Workman K. , Foster M. , Faber G.H. , Mair D.
Published: March 2003

BACKGROUND: One in every 1000 units of platelets is bacterially contaminated, which puts patients at risk for transfusion‐associated sepsis and death. However, there is currently no screening test in place to detect contaminated units. The use of commercially available multiple‐reagent urine dipsticks for this purpose was evaluated.

STUDY DESIGN AND METHODS: Platelet concentrates were inoculated with either sterile saline or suspensions of Staphylococcus aureus, Staphylococcus epidermidis, Bacillus cereus, Klebsiella pneumoniae, or Serratia marcescens to a final concentration of 50 colony‐forming units (CFU) per mL. The platelets were analyzed daily by the use of multiple‐reagent strips, quantitative culture, and glucometry.

RESULTS: B. cereus grew rapidly, reaching 107 CFU per mL 1 day after inoculation, while S. epidermidis grew slowly, achieving similar concentrations 4 to 6 days after inoculation. Two of 10 dipstick reagents, glucose and pH, proved useful in detecting bacteria. Both were lower in bacterially contaminated units than in controls. Glucose data obtained from automated analyzers validated the dipstick data. All organisms were detected at concentrations ≥107 CFU per mL, and S. aureus and K. pneumoniae were detected in the range of 103 to 105 CFU per mL.

CONCLUSION: The multiple‐reagent test used had a sensitivity and specificity of 95 percent (≥107 CFU/mL) and 98 to 100 percent, respectively. These data indicate that urine dipsticks can be used to rapidly and inexpensively detect bacterial contamination in platelet concentrates, which potentially will reduce morbidity and mortality at minimal cost.
At least three hepatitis C virus strains implicated in Swedish and Danish patients with intravenous immunoglobulin‐associated hepatitis C
Authors: Widell A , Zhang Y.‐Y. , Andersson‐Gäre B. , Hammarström L.
Published: March 2003

BACKGROUND: Three reported Swedish cases of hepatitis C in patients receiving an intravenous immunoglobulin (Gammagard, Baxter Healthcare, Deerfield, IL) were among the first to bring to light a worldwide outbreak of hepatitis C associated with non‐solvent/detergent (SD)‐treated Gammagard. In February 1994, all implicated batches of Gammagard were recalled and exposed patients traced.

STUDY DESIGN AND METHODS: Sera from all identified and hepatitis C‐viremic Swedish and Danish patients (n = 14) exposed to the implicated batches underwent hepatitis C virus genotyping and sequencing of the core region and hypervariable region 1 of E2. Genomic amplification was also done on 15 non‐SD‐treated batches of Gammagard.

RESULTS: Twelve patients were infected with subtype 1a and surprisingly, two with subtype 2b. Analysis of the core region showed identical sequences in four patients and the only consistently positive batch. Five patients shared another sequence, whereas three other subtype 1a patients each manifested unique sequences. The two subtype 2b isolates were identical. Genomic fingerprinting of the hypervariable region confirmed identity within each group with great stringency. Amplification with iso‐late‐specific primers showed mixed infection in one patient whose exposure was confined to a single batch.

CONCLUSION: The few batches implicated presumably were contaminated with several strains.
Inhibitory effect of 0 degree C storage on the proliferation of Yersinia enterocolitica in donated blood
Authors: Bradley Robert M. , Gander Rita M. , Patel Sujata K. , Kaplan Harold S.
Published: March 2003

BACKGROUND: Yersinia enterocolitica is frequently identified in cases of bacterial sepsis due to red cell transfusion. One of the features that makes Y. enterocolitica particularly dangerous is that, unlike most other bacterial contaminants of blood components, this organism can actively multiply in currently recommended refrigerator temperatures (1‐6 degrees C). The effect of a colder than normal storage temperature on Y. enterocolitica growth was investigated to determine whether bacteria growth could be reduced or inhibited at 0 degree C. STUDY DESIGN AND METHODS: Twenty‐four units of freshly collected donated blood were obtained. Three sets of 7 units each were inoculated with Y. enterocolitica O:3, Y. enterocolitica O:20, and Y. enterocolitica O:5, 27, respectively. The remaining 3 units served as uninoculated controls. Each of the 24 bags was split into two equal aliquots, with one aliquot stored at 4 degrees C and the other at 0 degree C. Bacteria growth was measured twice weekly for 6 weeks. Endotoxin and hemoglobin levels were also measured at selected intervals. RESULTS: Bacteria growth was detected earlier and in higher concentrations in the aliquots stored at 4 degrees C. Twenty‐two of the 42 inoculated aliquots had measureable bacteria growth. Thirteen aliquots had been maintained at 4 degrees C, and nine had been stored at 0 degree C. Sixteen of these 22 aliquots were matched pairs. Exponential growth was detected after 14 to 32 days in the 4 degrees C aliquots and after 28 to 39 days in the 0 degree C aliquots. Final bacteria counts were much higher in the 4 degrees C aliquots (10(5)‐ 10(14) colony‐forming units/mL) than in the 0 degree C aliquots (10(1)‐ 10(4) colony‐forming units/mL) on Day 42. Endotoxin was present in all 13 of the 4 degrees C aliquots with actively growing Y. enterocolitica. CONCLUSION: Storage of red cells at 0 degree C markedly prolongs the time required for Y. enterocolitica to achieve exponential grwoth and results in lower concentrations of bacteria.
Maternal immunization to Gov system alloantigens on human platelets
Authors: Bordin José O. , Kelton John G. , Warner Margaret N. , Smith James W. , Denomme Gregory A. , Warkentin Theodore E. , McGrath Katherine , Minchinton Robyn , Hayward Catherine P.M.
Published: March 2003

BACKGROUND: Immunization to platelet alloantigens can occur during pregnancy or after the transfusion of blood components. Platelet alloantibodies can cause neonatal alloimmune thrombocytopenia and posttransfusion purpura. Transfusion‐induced alloimmunization to a novel platelet alloantigen system, Gov, expressed on the 175‐kDa glycosyl phosphatidylinositol‐anchored platelet glycoprotein, CD109, was previously described. This report describes three unrelated patients who were alloimmunized to Gov(a) or Gov(b) during pregnancy. STUDY DESIGN AND METHODS: Platelets were typed by using radioimmunoprecipitation for HPA‐1a, −3a, −5a, −5b, Gov(a), and Gov(b) and by polymerase chain reaction‐restriction fragment length polymorphism for HPA‐1a, −1b, −3a, and −3b. Maternal sera were screened for platelet antibodies by using radioimmunoprecipitation and the antigen capture assay. RESULTS: Patients 1 and 2 were investigated after the diagnosis of neonatal alloimmune thrombocytopenia in their children, and alloantibodies specific for Gov(b) and Gov(a), respectively, were detected in maternal serum. Serum from patient 3, who had mild idiopathic thrombocytopenia purpura with no detectable autoantibody, was found to contain alloantibodies to Gov(b) and to HPA‐ 5b, presumably as a result of immunization during pregnancy. Platelet typings confirmed that the patients were at risk for alloimmunization to the respective antigen. CONCLUSION: This report of three cases of maternal alloimmunization to antigens in the Gov system indicates that immunization can occur via placental transfer of antigen and that Gov system alloantibodies may be associated with neonatal alloimmune thrombocytopenia.
Transfusion‐ and community‐acquired cytomegalovirus infection in children with malignant disease: a prospective study
Authors: Preiksaitis J.K. , Desai S. , Vaudry W. , Roberts S. , Akabutu J. , Grundy P. , Wilson B. , Boshkov L. , Hannon J. , Joffres M.
Published: March 2003

BACKGROUND: The use of cytomegalovirus (CMV)‐“safe” blood has been recommended for CMV seronegative patients with newly diagnosed malignant disease for whom bone marrow transplantation is a future option. STUDY DESIGN AND METHODS: To evaluate this policy, 76 CMV‐ seronegative children with lymphoreticular malignancies or solid tumors were randomly assigned to receive either blood components that were not screened for CMV antibody or CMV‐seronegative red cell (RBC) and platelet units. Subjects were followed for evidence of CMV infection by the use of enzyme‐linked immunosorbent assays and virus isolation. Follow‐up continued long after the blood transfusions to determine the risk of community‐acquired CMV infection. RESULTS: No cases of transfusion‐acquired CMV infection were documented. The prevalence of CMV IgG and IgM antibody in blood donors was 40.5 and 0.9 percent, respectively. Patients assigned to receive standard blood components and CMV‐negative components were given a median (range) of 7 (1‐30) and 9 (1‐38) RBC units and 11 (0‐123) and 14 (0‐71) platelet units, respectively. The risk of transfusion‐acquired CMV infection is estimated to be less than 1 in 698 donor exposures. Two patients developed asymptomatic community‐acquired CMV infection, for an incidence of 1.7 percent per patient‐year of follow‐up. CONCLUSION: The risk of transfusion‐acquired CMV infection in this population is low, largely because of the patients' low level of exposure to seropositive blood and the use of relatively white cell‐reduced components for purposes other than CMV prevention. Such children at this center therefore continue to receive standard blood components. Strategies to prevent CMV seroconversion in these children should include parental education to minimize the risk of community‐acquired infection.
Difficulties in identifying antibodies in the Dombrock blood group system in multiply alloimmunized patients
Authors: Strupp A. , Cash K. , Uehlinger J.
Published: March 2003

BACKGROUND: Although red cell (RBC) antibodies of the Dombrock blood group system have been reported to cause acute and delayed hemolytic transfusion reactions, the difficulty in identifying these antibodies in patients with multiple RBC alloantibodies has not previously been discussed. The cases of four sickle cell disease patients who developed Dombrock system antibodies after transfusion, three of which were discovered in association with hemolytic transfusion reactions, are reported. CASE REPORTS: Patient 1 was a 36‐year‐old woman with multiple RBC alloantibodies. Because of the lack of an increment in hematocrit after transfusion, an investigation was performed; it revealed anti‐ Do(b) in the serum. Patient 2 was a 30‐year‐old woman with known anti‐ C, ‐E, ‐K, ‐S, ‐Fya, and ‐Bga. She had received a transfusion 10 days previously. Before further transfusion was begun, antibody identification revealed weak nonspecific reactions, which were thought to be HLA antibodies. She developed acute hemolysis during RBC exchange for acute chest syndrome; anti‐Doa was identified in both the serum and eluate. She received 2 units of Do(a‐) RBCs without complication. Patient 3 was a 35‐year‐old woman with known anti‐C, ‐E, ‐K, ‐Fya, and ‐ N and a warm autoantibody. Two weeks after transfusion, she had a delayed hemolytic transfusion reaction coincident with the identification of anti‐Doa. Patient 4 was a 33‐year‐old woman with known anti‐C, ‐V, ‐K, ‐Fya, ‐Fy3, Jkb, ‐S, ‐N, and ‐Ytb, who developed anti‐Doa 8 weeks after transfusion. CONCLUSION: An association of Dombrock blood group system antibodies with hemolytic reactions is demonstrated in alloimmunized sickle cell disease patients. In all four cases, identification of Dombrock system antibodies was delayed because high‐titer low‐avidity antibodies, HLA antibodies, or autoantibodies were thought to explain the serologic findings. The presence of Dombrock system antibodies should be considered when unexplained serologic reactivity occurs during antibody identification in this population.
White cell apoptosis in packed red cells
Authors: Frabetti F. , Musiani D. , Marini M. , Fanelli C. , Coppola S. , Ghibelli L. , Tazzari P. L. , Bontadini A. , Tassi C. , Conte R.
Published: March 2003

BACKGROUND: After the removal of the buffy coat, packed red cell (RBC) transfusion units still contain white cells that may undergo apoptosis as a result of storage conditions (1‐6 degrees C). The aim of the present study was the evaluation of this phenomenon in view of the possible influence it may have on febrile nonhemolytic transfusion reactions. STUDY DESIGN AND METHODS: Three independent methods (microscopy, DNA electrophoresis, and cytometry) were used to evaluate apoptosis in white cells present in 13 RBC units. Of these units, 10 had been collected into CPD/saline‐adenine‐glucose‐mannitol and 3 into CPDA‐1; each bag was split in two parts, one of which was irradiated. RBCs were stored at 1 to 6 degrees C, and samples were periodically withdrawn for study. The proliferative capacity of stored lymphocytes was evaluated after phytohemagglutinin stimulation and tritiated thymidine incorporation. RESULTS: Apoptosis was found to occur in both granulocytes and lymphocytes, starting from the first 48 to 72 hours of storage. The choice of the anticoagulant‐preservative solution and the effect of irradiation did not influence the amount and the timing of the apoptotic phenomenon. Lymphocyte proliferative capacity was found to decrease sharply with storage time. CONCLUSION: Conditions of storage in RBCs induce consistent apoptosis in residual white cells. The possible clinical implications of the relationships between apoptosis and the induction of biologic response modifiers (that may cause interleukin‐mediated febrile non‐hemolytic transfusion reactions) and between apoptosis and immune reactions remain to be elucidated.
No‐fault compensation for transfusion‐associated hepatitis B virus, hepatitis C virus, and HIV infection: Italian law and the Tuscan experience
Authors: Fineschi V. , Cateni C. , Fanetti P. L. , Turillazzi E.
Published: March 2003

BACKGROUND: On February 25, 1992, in Italy, a law (Number 210; referred to as 210/1992) was promulgated providing economic indemnity for persons infected with hepatitis B virus, hepatitis C virus, and HIV via transfusion or the administration of hemoderivatives.

STUDY DESIGN AND METHODS: The requests for compensation presented in the central Italian region of Tuscany from the time of the law's promulgation through December 31, 1996, were analyzed. These requests are surveyed by medical commissions in the regional military hospitals, which must compile a report of the completed assessments, formulate a decision concerning verified illnesses, and express an opinion on the existence of a relationship of causality between the damaging event and the impairment or death of the subjects.

RESULTS: Out of 428 requests for indemnity, 372 have been granted and 56 denied. Posttransfusion infections (286 cases) were clearly more prevalent than those due to hemoderivatives (141 cases). Cases of hepatitis, particularly type C, constitute the great majority of the infections for which indemnity was sought, while cases of HIV infection are scarce and in sharp decrease in the data from 1995 and 1996.

CONCLUSIONS: Italian Law 210/1992 has been recognized as providing a benefit to persons infected medically, but its efficiency is greatly obstructed by a lack of documentation regarding transfusions performed in the past.
Exposure to GB virus type C or hepatitis G virus in selected Australian adult and children populations
Authors: Hyland C. A. , Mison L. , Solomon N. , Cockerill J. , Wang L. , Hunt J. , Selvey L. A. , Faoagali J. , Cooksley W. G. E. , Young I. F. , Trowbridge R. , Borthwick I. , Gowans E. J.
Published: March 2003

BACKGROUND: The epidemiology and disease association for the GB virus type C (GBV‐C) or hepatitis G virus (HGV) are poorly understood.

STUDY DESIGN AND METHODS: This study describes the exposure rates to GBV‐ C/HGV in diverse Australian population groups by testing for current infection and evidence of past infection with a reverse transcriptase polymerase chain reaction and an anti‐E2 enzyme‐linked immunosorbent assay, respectively. Subjects included volunteer blood donors, hepatitis C antibody (anti‐HCV)‐positive donors, children, hemodialysis patients, pregnant women attending a prenatal clinic, injecting drug users (IVDUs), and adult hemophiliacs.

RESULTS: Combined GBV‐C RNA and E2 antibody prevalence was 6.5 percent (6/93) in children, 13.3 percent (75/565) in blood donors, 14 percent (14/99) in pregnant women, 22.5 percent (18/80) in hemodialysis patients, 80 percent (56/70) in anti‐ HCV‐positive donors, 88.6 percent (31/35) in IVDUs, and 85.7 percent (54/63) in adult hemophiliacs. Children had the lowest antibody rate, 1.1 percent, whereas the rate was 10.8 percent for blood donors and rose to 45.7 percent for IVDUs, 57.1 percent for anti‐HCV‐positive donors, and 74.6 percent for hemophiliacs. In contrast, current infection rates were comparable for children, blood donors, and pregnant women (5.4, 2.6, and 6%, respectively), rising to 11.1 percent for hemophiliacs, 24.3 percent for anti‐HCV‐positive donors, and 48.6 percent for IVDUs. Ten of 12 blood donors had persistent viremia, while 2 had recent infections, 1 with apparent resolution.

CONCLUSION: Exposure to GBV‐C can commence at an early age, although ongoing exposure may also occur among adults with no apparent risk factors. GBV‐ C RNA positivity was not associated with abnormal plasma alanine aminotransferase levels among blood donors.
Transfusion of buffy coat‐depleted blood components and risk of postoperative infection in orthopedic patients
Authors: Innerhofer P. , Walleczek C. , Luz G. , Hobisch‐Hagen P. , Benzer A. , Stöckl B. , Hessenberger G. , Nussbaumer W. , Schobersberger W.
Published: March 2003

BACKGROUND: Allogeneic blood transfusions have been reported to increase susceptibility to postoperative infection, but the findings were inconclusive. This study was designed to investigate the effect of buffy coat‐depleted allogeneic and autologous transfusion on postoperative infection in patients undergoing orthopedic surgery.

STUDY DESIGN AND METHODS: Patients (n = 385) undergoing elective orthopedic surgery (primary and revision joint replacement, spinal, or pelvic surgery) were included in a prospective observational study of the incidence of postoperative infection between April and December 1996. Infection rates in patients who received allogeneic buffy coat‐depleted blood transfusions were compared with those in patients who received no transfusion or only autologous (buffy coat‐depleted) blood.

RESULTS: Patients without exposure to allogeneic blood (no blood or only autologous blood) had an infection rate of 3.9 percent, as compared to a rate of 12.2 percent for those with exposure to allogeneic blood (allogeneic blood, autologous plus allogeneic blood) (odds ratio 3.442; 95% CI, 1.349‐10.40; p = 0.006). Of the 385 study patients, 309 underwent primary hip or knee replacement surgery. In this homogeneous subgroup, the postoperative infection rate was 4.6 percent after no transfusion or autologous transfusion and 11.9 percent after allogeneic transfusion (odds ratio 2.827; 95% CI 1.059‐8.799; p = 0.036). Multivariate regression analysis confirmed buffy coat‐depleted allogeneic blood transfusion as an independent variable associated with high risk for postoperative infection.

CONCLUSION: Buffy coat‐depleted allogeneic blood transfusion increases the incidence of postoperative infection in patients undergoing uncontaminated orthopedic surgery.
Effects of granulocyte–colony‐stimulating factor on potential normal granulocyte donors
Authors: McCullough J. , Clay M. , Herr G. , Smith J. , Stroncek D.
Published: March 2003

BACKGROUND: The use of granulocyte–colony‐stimulating factor (G–CSF) to increase the granulocyte count and the yield from leukapheresis in normal donors is leading to renewed interest in granulocyte transfusion. Therefore, it is important to understand the side effects of G–CSF.

STUDY DESIGN AND METHODS: We studied the effect of G–CSF on peripheral blood counts and recorded the side effects experienced 24 hours after an injection of G–CSF in normal subjects donating peripheral blood progenitor cells for research.

RESULTS: Following administration of G–CSF to 261 donors, the neutrophil count increased to 20.6 to 24.5 × 109 per μL depending on the dose of G–CSF. This represented a 6.2 to 7.4‐fold increase over the neutrophil count before G–CSF administration. Of all donors, 69 percent experienced one or more side effects. The most common effects were: muscle and bone pain, headache, fatigue, and nausea. There was a relationship between the dose of G–CSF and the likelihood of experiencing a side effect. Most side effects were mild, but about 75 percent of donors took analgesics because of them.

CONCLUSIONS: In a granulocyte donation program involving G–CSF stimulation, about two‐thirds of donors would experience one or more side effects, but these would usually be mild and well tolerated.
Revisiting the issue: can the reading for serologic reactivity following 37°C incubation be omitted?
Authors: Judd W. John , Fullen Douglas R. , Steiner E. Ann , Davenport Robertson D. , Knafl Pamela C.
Published: March 2003

BACKGROUND: Omitting the 37°C reading from screening tests for unexpected antibodies results in failure to detect some Rh, K, and Jk agglutinins of potential significance (wanted positives). However, this measure avoids unwanted positive tests due to cold agglutinins.

STUDY DESIGN AND METHODS: Using data from prior publications, actual risk calculations (ARCs) were made to predict the risk of eliminating the 37°C reading, pretransfusion direct antiglobulin test (DAT), and routine indirect antiglobulin crossmatch (IAT‐XM). ARCs used the equation: wanted positives missed × 0.34 (or 0.80) × 5 × percent antigen‐positive, where 0.34 = percent of patients transfused (ARCs for 37°C reading and DAT); 0.80 = percent of crossmatched patients transfused (ARCs for IAT‐XM); 5 = average number of units transfused. Following elimination of the 37°C reading, the impact of this change on patient care was monitored. Antibody detection and identification data and transfusion reaction reports for 6 months after the change were reviewed. Recently transfused patients with new antibodies were evaluated for immune hemolysis by review of clinical and laboratory data. The findings were compared with those from the same dates of the preceding year.

RESULTS: The risk of transfusing incompatible blood by eliminating the DAT, IAT‐XM, and 37°C reading is approximately 1:13,000, 1:2,000, and 1:2,400 units transfused, respectively. The cumulative risk from eliminating all three tests is approximately. 1:1,000 units. With respect to the 37°C reading, there were no differences between the pre‐change and post‐change study periods in the incidence of reported transfusion reactions or cases of immune hemolysis associated with newly formed antibodies. However, unwanted positive tests decreased from 162 to 61 following elimination of the 37°C reading. This represents a decrease of 20 percent in the number of samples requiring antibody identification annually.

CONCLUSIONS: Eliminating the 37°C reading from pretransfusion antibody screening tests imposes less risk than omitting the routine IAT‐XM, and it avoids the time and costs of evaluating unwanted positive tests, thus reducing expenditures and delays in patient care.
Retroviral transduction and expansion of peripheral blood lymphocytes for the treatment of mucopolysaccharidosis type II, Hunter's syndrome
Authors: Stroncek David F. , Hubel Allison , Shankar Raji A. , Burger Scott R. , Pan Dao , McCullough Jeffrey , Whitley Chester B.
Published: March 2003

BACKGROUND: Gene therapy using autologous peripheral blood lymphocytes (PBLs) has been used to produce adenosine deaminase with which to treat patients with severe combined immunodeficiency. Patients with mucopolysaccharidosis type II (MPS II) lack iduronate‐2‐sulfatase (IDS), and serial PBL gene therapy may benefit these patients.

STUDY DESIGN AND METHODS: The purpose of these studies was to develop a method to transduce PBLs from a patient with MPS II by using a retroviral vector, LS2N, containing the IDS gene. PBLs were collected by apheresis and cryopreserved in aliquots for the performance of multiple transductions and expansions. The PBLs were expanded in number and then transduced in a hollow‐fiber bioreactor (HFBR). Additional culture allowed for further expansion.

RESULTS: Fresh PBLs (6.2 × 107) from a patient with MPS II were transduced with L2SN and expanded in an HFBR with an extracapillary space of 11 mL. After 10 days of culture, 4.1 × 109 cells were harvested. Cryopreserved MPS II PBLs could not be reliably expanded if they were placed in the HFBR immediately after being thawed; however, cells were successfully transduced and expanded in the HFBR if they were first cultured in a bag. To increase the cell yield, PBLs were expanded in a 60‐mL HFBR after transduction and expansion in an 11‐mL HFBR. In four separate experiments, 2 × 108 cryopreserved PBL were cultured for 3 days in a bag and transferred to an 11‐mL HFBR, where they were transduced daily with L2SN for 3 days and then expanded for 4 additional days. Cells were then transferred into a 60‐mL HFBR and expanded for an additional 7 days. In the four experiments, 5.5 × 109, 7.4 × 109, 1.12 × 109, and 19.4 × 109 cells were produced. The vector was detected in the harvested cells, but the proportion of cells transduced was less than 2.5 percent, the lowest standard used in the assay. In two of the experiments, cells harvested from the HFBR were used in a gene therapy clinical trial.

CONCLUSION: Autologous cryopreserved PBLs can be transduced and expanded to produce >1 × 1010 cells. This procedure is being used for a Phase I/II clinical trial of lymphocyte gene therapy.