bluuebunny/crossref_metadata_2025_split huggingface dataset:
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.
DNA from blood samples can be used to genotype patients who have recently received a transfusion
Authors: Reid M.E. , Rios M. , Powell V.I. , Charles‐Pierre D. , Malavade V. 
Published: March 2003

BACKGROUND: The use of hemagglutination to phenotype red cells from recently transfused patients or of red cells that are coated with IgG can be time‐consuming and difficult to interpret. Because the molecular bases of many blood group antigens are known, it was investigated whether polymerase chain reaction (PCR) analysis of DNA, from white cells in blood from transfused patients, could be used to predict the blood group antigen profile of a patient.STUDY DESIGN AND METHODS: To prevent problems arising from potentially poor‐quality DNA in clinical samples, primers that flanked the polymorphism of interest and that replicated a relatively short PCR amplicon were used. The PCR products, with or without digestion with the appropriate restriction enzyme, were analyzed on gels. Samples were collected from 60 patients who had received from 2 to 50 units of RBCs in the 7 days before sample collection. RBCs from some of these patients were coated with IgG. Analyses for RHD/non‐D, RHE/RHe, KEL1/KEL2, FYA/FYB, FY‐GATA, JKA/JKB, and GYPA M/N were performed by using assays that had been validated with DNA prepared from untransfused volunteers of known phenotype. The genotyping assays were performed without knowledge of the expected result.RESULTS: The predicted genotype after analysis of the 60 patient samples was that expected from the results of phenotyping. In all cases, the molecular analysis gave a single result; no evidence of chimerism was obtained.CONCLUSION: In each case, the molecular genotype results were in agreement with the blood group antigen as determined by historical phenotyping, phenotyping after hypotonic washing, detection of alloantibodies in the patient's serum, or elution of alloantibody(ies). Under the conditions of these assays, reliable determination of a blood group allele can be made by PCR‐restriction fragment length polymorphism testing.
Diversion of initial blood flow to prevent whole‐blood contamination by skin surface bacteria: an in vitro model
Authors: Wagner S.J. , Robinette D. , Friedman L.I. , Miripol J. 
Published: March 2003

BACKGROUND: Sepsis arising from the transfusion of bacterially contaminated platelet components continues to be an infrequent, yet serious transfusion complication. Skin organisms are implicated in a number of these septic episodes. A model system was used to investigate if a skin organism's bioburden in blood components could be reduced by diverting the first few mL of whole blood away from the primary container.STUDY DESIGN AND METHODS: A sterile medication site was inserted into a bag containing sterile saline or whole blood; the site was deliberately contaminated with Staphylococcus aureus and allowed to dry. After needle puncture of the contaminated medication site, bacteria levels were measured 1) in successive 7‐mL tubes of blood or saline drawn through a diversion arm, 2) in 40 mL of a connected transfer pack, and 3) in blood or saline from a needle puncture of the original container via another sterile medication port.RESULTS: Diverting the first 21 to 42 mL of saline or whole blood reduces the downstream bioburden of deliberately introduced surface S. aureus by approximately 1 log.CONCLUSION: Development of a diversion system for collection of whole blood in sample tubes before filling the primary container may reduce the bioburden of subsequently prepared components and thereby the frequency of sepsis due to skin organisms.
Irradiation of platelet components: inhibition of lymphocyte proliferation assessed by limiting‐dilution analysis
Authors: Luban N.L.C. , Drothler D. , Moroff G. , Quinones R. 
Published: March 2003

BACKGROUND: Using a limiting‐dilution analysis (LDA) assay that measures clonigenic T cells, it has been demonstrated that, with 2500 cGy, there is no T‐cell growth in red cell components irradiated in blood bags. In the current study, the LDA assay was used to investigate the effect of gamma radiation on the proliferative capacity of T cells in plateletpheresis components.STUDY DESIGN AND METHODS: Platelets were collected by using an apheresis instrument and settings that provided sufficient mononuclear cells for the LDA assay. Platelet components (n = 8) were irradiated in 1‐L plastic bags 24 hours after collection with 500, 1500, and 2500 cGy of gamma radiation in a stepwise manner. Mononuclear cells were isolated after each irradiation dose by the use of ficoll‐hypaque. A density separation medium was used to reduce the platelet numbers. T cells were enumerated by fluorescence‐activated cell sorter and functionally assessed by LDA assay, which quantified T cells proliferating in the presence of polyclonal stimuli and cytokines. The frequency of T‐cell growth (f) was visually scored after 4 weeks of incubation at 37°C. Data were calculated as fexperimental/fcontrol and expressed as log10 reduction.RESULTS: The T‐cell content of the mononuclear cell population was 17 ± 10.5 percent, which was unaltered by irradiation. After 500‐cGy irradiation, functional T cells were reduced by 2.09 log10. Irradiation with 1500 cGy resulted in a 3.96 log10 reduction, but viable clonable T cells were detected in all experiments. With 2500‐cGy irradiation, no T‐cell growth was detected; this represented a greater than 4.86 log10 reduction.CONCLUSION: As the dose of gamma radiation delivered to plateletpheresis components increased, the number of residual functional T cells decreased exponentially. Irradiation with 2500 cGy inactivates T cells in apheresis platelets, as measured by an LDA assay.
Antibiotic‐labeled probes and microvolume fluorimetry for the rapid detection of bacterial contamination in platelet components: a preliminary report
Authors: Brecher M.E. , Wong E.C.C. , Chen S.E. , Vampola C. , Rocco R.M. 
Published: March 2003

BACKGROUND: Approximately 1 platelet in 2000 components is bacterially contaminated. Most commonly, contaminating organisms are gram positive skin saprophytes (such as Staphylococcus sp. or Bacillus sp.). A novel approach to the rapid diagnosis of gram positive contamination by the use of a fluorescence‐labeled antibiotic probe with affinity for the gram positive cell was investigated.STUDY DESIGN AND METHODS: Two isolates of Staphylococcus epidermidis were inoculated into bags of Day 0 platelets. Quantitative cultures along with a semi‐automated screening assay on a microvolume fluorimeter employing a fluorescence‐conjugated vancomycin probe was performed for each day of storage. In addition, serial dilutions of the bacteria were added to sterile platelets to achieve a range spanning 101 to 108 CFUs per mL.RESULTS: All samples with a bacterial contamination of ≥105 CFU per mL were detected. Sterile samples were nonreactive. The entire procedure requires three pipetting steps and took less than 1 hour to perform.CONCLUSION: These preliminary results with the use of fluorescence‐labeled antibiotics as probes combined with microvolume fluorimetry for the rapid detection of bacterial contamination of platelet components suggest that this is a promising approach. Further studies with additional organisms and alternative conjugates, bacteria, and antibiotics are underway.
Molecular analysis of HCV type 1 to 5 envelope gene: application to investigations of posttransfusion transmission of HCV
Authors: Cantaloube J.‐F. , Venault H. , Zappitelli J.‐P. , Gallian P. , Touinssi M. , Attoui H. , Biagini P. , De Lamballerie X. , De Micco P. 
Published: March 2003

BACKGROUND: Until 1990, HCV infection was common in transfused patients, resulting in more than 200,000 cases of posttransfusion hepatitis C in France alone. A molecular method that permits the investigation of posttransfusion hepatitis C infections is presented.STUDY DESIGN AND METHODS: Viral sequences in the envelope region of HCV were obtained for 12 pairs of blood recipients and their respective blood donors. The HCV strains studied belonged to types 1 (subtypes 1a and 1b), 2, 3, 4, and 5. Genetic distances and mutation rates were determined, and sequences were submitted to phylogenetic analysis along with sequences retrieved from nucleotide databases.RESULTS: Pairwise distances in the donor‐recipient pairs were found to be less than 0.05 mutation per site, which corresponds to a mutation rate ranging from 0.6 × 10−3 to 2.1 × 10−3 per site per year. Sequences obtained from the 12 donor‐recipient pairs clustered in 12 monophyletic nests.CONCLUSION: The genetic analysis of the envelope region of HCV can be used for the forensic evaluation of virus transmission. It permits the refutation of a link between blood transfusion and HCV transmission, rather than proof of the existence of such a link.
Immunologic changes after transfusion of autologous or allogeneic buffy coat‐poor versus WBC‐reduced blood transfusions in patients undergoing arthroplasty.II. Activation of T cells, macrophages, and cell‐mediated lympholysis
Authors: Innerhofer P. , Tilz G. , Fuchs D. , Luz G. , Hobisch‐Hagen P. , Schobersberger W. , Nussbaumer W. , Lochs A. , Irschick E. 
Published: March 2003

BACKGROUND: To estimate the impact of RBC preparations on the status of postoperative immune activation, the soluble cytokine receptors of TNFα (sTNF‐R) and IL‐2 (sIL‐2R), as well as neopterin and cell‐mediated lympholysis (CML), were measured.STUDY DESIGN AND METHODS: Patients undergoing strictly standardized anesthesiologic management for elective orthopedic surgery were enrolled in a prospective study. The perioperative course (Days 0, 3, 7, and 10) of sTNF‐R, sIL‐2R, neopterin, and CML was compared after random assignment to allogeneic buffy coat‐reduced (Group 2, n = 8) or WBC‐reduced (Group 3, n = 11) RBC transfusion regimen. Recipients of autologous buffy coat‐reduced RBC transfusions (Group 1, n = 15) served as controls. Patients receiving intraoperatively and postoperatively salvaged blood only (n = 10) were separately analyzed as Group 4.RESULTS: In Group 1, a short‐lasting increase in soluble cytokine receptors, a diminished cytolytic response (Day 0 vs. Day 7: sTNF‐R, p = 0.0001; sIL‐2R, p = 0.0004; CML, p = 0.0238), and an elevation of neopterin (Day 0 vs. Day 3: p = 0.0064) were observed. In contrast, in allogeneically transfused patients, sTNF‐R (Group 2, p = 0.0469: Group 3, p = 0.0039), sIL‐2R (Group 3, p = 0.002) and neopterin (Group 3, p = 0.0164) increased further from baseline to Day 10 (Day 0 vs. Day 10), and this increase was accompanied by a diminished cytolytic response (Day 0 vs. Day 10: Group 2, p = 0.05; Group 3, p = 0.0076). Patients in Group 4 showed a short‐lasting increase in sIL‐2R (Day 0 vs. Day 3: p = 0.0078), neopterin (Day 0 vs. Day 3: p = 0.0156) and sTNF‐R (Day 0 vs. Day 7: p = 0.0781).CONCLUSION: Allogeneic transfusions seem to prolong the postoperative status of immune activation, even when WBC‐filtered RBCs are used for the transfusion regimen.
The mechanism of apoptosis in human platelets during storage
Authors: Li Junzhi , Xia Yuping , Bertino Amy M. , Coburn John P. , Kuter David J. 
Published: March 2003

BACKGROUND: Although it is usually involved only in nucleated cells (NCs), artificially enucleated cells also lose viability by a programmed process of cell death called apoptosis. Because platelets undergo loss of viability during storage, an attempt was made to determine whether platelets contained the apoptotic mechanisms and whether it was activated during platelet storage.STUDY DESIGN AND METHODS: Platelet viability was measured by reduction of a tetrazolium dye (MTS) and annexin V binding. Members of the death receptor, caspase, and Bcl‐2 families were detected by RNase protection assay and Western blotting. Caspase 3 activation was measured by enzyme and Western blot assays and by cleavage of gelsolin.RESULTS: After 5 days of storage under standard blood banking conditions, platelets display biochemical signs of apoptosis by losing MTS activity and increasing the amount of phosphatidylserine on their surface. The mRNA and the proenzyme for several members of the caspase, death receptor, and Bcl‐2 families are expressed at high levels in platelets. An increase in caspase 3 activity and the amount of the biologically active p17 subunit of active caspase 3 were observed to coincide with the appearance of apoptotic markers during storage. These effects were not due to platelet activation. The caspase 3 substrate, gelsolin, began to undergo proteolysis after 3 to 4 days of storage, and the addition of the caspase inhibitor z‐VAD‐fmt substantially inhibited this process.CONCLUSION: Platelets contain many of the components of the apoptotic mechanism and show activation of caspase 3 and consequent cleavage of gelsolin during storage, independent of platelet activation. Evaluation of the mechanism of apoptosis in platelets may provide a basis for developing novel strategies to enhance platelet viability during storage.
Filtration of methylene blue‐photooxidized plasma: influence on coagulation and cellular contamination
Authors: Riggert Joachim , Humpe Andreas , Legler Tobias J. , Wolf Cornelia , Simson Georg , Köhler Michael 
Published: March 2003

BACKGROUND: Virus inactivation of plasma can be achieved by photodynamic methods in the presence of phenothiazine dyes such as methylene blue (MB). Subsequent filtration may increase the efficacy of virus inactivation and reduce adverse effects of WBC contamination and MB.STUDY DESIGN AND METHODS: This study examined the effect of filtration with three different filters (MBF1, MBF2, and MBF3) on MB concentration, residual cells, coagulation factors, and activation measures of coagulation, fibrinolysis, and complement in MB‐treated (1 μM/L) plasma units.RESULTS: Filtration reduced the concentration of MB by ≥89 percent. WBCs were depleted by 92 percent (MBF1) and >99.9 percent (MBF2 and MBF3). Treatment with MB significantly decreased the coagulation potency from levels in untreated plasma, as measured by thromboplastin time ratio (112 ± 18% vs. 95 ± 11%), activated partial thromboplastin time (40 ± 3 sec vs. 44 ± 3 sec), thrombin time (16.9 ± 1.1 sec vs. 18.6 ± 1.5 sec), factor VIII (1.09 ± 0.21 U/mL vs. 0.85 ± 0.13 U/mL), and vWF (0.94 ± 0.65 U/mL vs. 0.65 ± 0.24 U/mL). Filtration did not further decrease these values, while factor XI (0.75 ± 0.22 U/mL vs. 0.37 ± 0.20 U/mL) and prekallikrein values decreased in MB plasma units filtered with the MBF3. In addition, activated factor XII (0.7 ± 0.5 μg/L vs. 4.5 ± 1.0 μg/L) increased.CONCLUSION: WBCs and MB can be eliminated from MB‐treated plasma units by filtration. Differences in biocompatibility of the different filters, especially the influence on the contact phase of coagulation, must be taken into consideration.