bluuebunny/crossref_metadata_2025 huggingface dataset:
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.
Collection of MNCs with two cell separators for adoptive immunotherapy in patients with stage IV melanoma
Authors: Glaser Anke , Schuler‐Thurner Beatrice , Feuerstein Bernadette , Zingsem Jurgen , Zimmermann Robert , Weisbach Volker , Eckstein Reinhold
Published: March 2003

BACKGROUND: MNCs for adoptive immunotherapy may be collected by leukocytapheresis with a cell separator.

STUDY DESIGN AND METHODS: Six healthy cytapheresis donors donated two MNC concentrates on a cell separator (AS.TEC 204, Fresenius): one on the standard MNC program and one on a modified MNC program with reduced centrifuge velocity that leads to a lower platelet contamination. Seventeen patients with malignant melanoma donated 26 MNC concentrates: 5 on the AS.TEC 204 MNC program, 9 on the modified AS.TEC 204 MNC program, and 12 on another modified MNC program (Spectra, COBE).

RESULTS: In the course of cultivation of MNCs to dendritic cells (DCs), the donor MNC concentrates with the lower platelet contamination (475 ± 85 × 109/L) had a significantly higher relative DC yield (low platelet contamination: 3.9 ± 1.6% of the plated cells; high platelet contamination: 2.5 ± 1.8% of the plated cells; p = 0.019) than the concentrates with the higher platelet contamination (2364 ± 448 × 109/L). No significant difference was found in the yields of MNCs and CD14+ cells in the three protocols used for the collection of MNCs from patients with melanoma. The components obtained by the standard AS.TEC 204 MNC program had a significantly higher platelet contamination (1768 ± 994 × 109/L) than the components obtained by the modified AS.TEC MNC program (360 ± 98 × 109/L; p<0.05) and the modified Spectra MNC program (636 ± 266 × 109/L); p<0.05). Because of the low number of investigated components, no significant difference in the DC yield of the three protocols could be detected (mean DC yield after cultivation: 746 ± 429 × 106).

CONCLUSION: A high platelet contamination of MNC concentrates intended for adoptive immunotherapy can lead to a significant impairment of the DC yield after cultivation. Both the modified AS.TEC 204 and the modified Spectra MNC programs are well suited for collecting MNC concentrates with high MNC yields and low platelet contamination from patients with malignant melanoma.
Neonatal alloimmune thrombocytopeniadue to anti‐Naka
Authors: Kankirawatana Suthida , Kupatawintu Pavinee , Juji Takeo , Veerakul Gavivann , Ngerncham Sopapan , Chongkolwatana Viroj , O'Charoen Rachanee
Published: March 2003

BACKGROUND: The accurate diagnosis of neonatal alloimmune thrombocytopenia is essential in the effective treatment of potentially serious bleeding in neonates.

CASE REPORT: Reported here is a case of a full‐term female baby who was delivered by vacuum extraction from a gravida 1 para 1 healthy mother. She presented with generalized petechiae and bilateral cephalhematoma, which she had had since birth. At 7 hours of life, she had an upper gastrointestinal hemorrhage and was found to have severe anemia and marked thrombo‐cytopenia. Coagulation screening tests were normal. The diagnosis of neonatal alloimmune thrombocytopenia was suspected, and maternal serum was collected for further study. The baby was treated with a single dose of hydrocortisone (10 mg/kg) and IVIG (400 mg/kg) while waiting for irradiated platelets from her mother. After 30 mL of a transfusion of maternal platelets, the baby's platelet count rose dramatically, from 15,000 to 162,000 per μL, and it remained stable at that level. She was discharged on the 10th hospital day in good condition. During the follow‐up period of 8 months, her growth and development were satisfactorily normal, as well as her platelet count. A high‐titered platelet antibody was detected in the maternal serum by use of a solid phase platelet adherence technique.

RESULTS: The specificity of the platelet antibody was identified as anti‐Naka by the mixed passive hemagglutination test method.

CONCLUSION: These findings suggested a diagnosis of NAIT caused by anti‐Naka.
Immune response to autologous transfusion in healthy volunteers: WB versus packed RBCs and FFP
Authors: Frietsch Thomas , Fessler Heiko , Kirschfink Michael , Nebe Thomas , Waschke Klaus F. , Lorentz Arnulf
Published: March 2003

BACKGROUND: Storage of blood as packed RBCs and FFP is standard practice in allogeneic transfusion. Separation into components has been proposed for autologous transfusion, as well, but beneficial effects have not yet been shown.

STUDY DESIGN AND METHODS: Twenty‐four healthy male volunteers were randomly assigned to receive 1 unit of either autologous RBCs and FFP (RCP group) or WB (WB group) after 49 or 35 days of storage, respectively. The immune response was analyzed by ELISA for IL‐6, C3a, terminal complement complex SC5b‐9, TNF‐α, and neopterin. Differential WBC counts and the phagocytosis of neutrophils and monocytes were measured by flow cytometry.

RESULTS: Cell counts of monocytes (0.85 × 103 ng/mL) and neutrophils (6.9 × 103 ng/mL) increased 30 minutes after WB transfusion and then returned to close to the baseline values seen in the RCP group (0.47 and 2.9 × 103 ng/mL, respectively) throughout the monitored period (p<0.05). C3a (169 vs. 116 ng/mL) and IL‐6 (29 vs. 6 pg/mL) reached higher plasma concentrations in the WB group (n = 11) than in the RCP group (n = 10). Phagocytosis of opsonized Escherichia coli was increased in neutrophils and monocytes and lasted up to 7 days after the transfusion of whole blood.

CONCLUSION: Autologous WB induces a modest immunomodulation, but this effect is not observed upon transfusion of autologous blood components.
Evaluation of an automated culture systemfor detecting bacterial contamination of platelets: an analysis with 15 contaminating organisms
Authors: Brecher Mark E. , Means Norman , Jere Charles S. , Heath David , Rothenberg Steve , Stutzman Les C.
Published: March 2003

BACKGROUND: Approximately 1 in 2000 platelet components are bacterially contaminated. The time to detection of 15 seeded organisms in platelets recovered from an automated culture system was studied.

STUDY DESIGN AND METHODS: Isolates of Bacillus cereus, Bacillus subtilis, Candida albicans, Clostridium perfringens, Corynebacterium species, Enterobacter cloacae, Escherichia coli, Klebsiella oxytoca, Propionibacterium acnes, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, Serratia marcescens, Streptococcus pyogenes, and Streptococcus viridans were inoculated into Day 2 apheresis platelet components to obtain a final concentration of approximately 10 and 100 CFU per mL (2 units/organism). Each bag was sampled 10 times (20 mL/sample). Four mL of each sample was inoculated into standard aerobic and anaerobic bottles and into aerobic and anaerobic bottles containing charcoal; 2 mL was inoculated into pediatric aerobic bottles (so as to maintain a 1:10 ratio of sample to media) and 1 mL into thioglycollate broth.

RESULTS: With the exception of P. acnes, all organisms were detected in a mean of 9.2 to 25.6 hours. A range of 10 serial dilutions in inoculating concentrations was associated with an overall 10.1‐percent difference in detection time. A mean of 74.4 and 86.2 hours (100 and 10 CFU/mL inocula, respectively) was required for the detection of P. acnes in anaerobic bottles.

CONCLUSION: Bacteria thought to be clinically significant platelet contaminants can be detected in 9.2 to 25.6 hours when the starting concentration is approximately 10 to 100 CFU per mL. P. acnes required considerably longer incubation times for detection (in either aerobic or anaerobic bottles). However, P. acnes is of questionable clinical significance. Such a detection system could be used in either a blood collection center or a transfusion service to screen platelet concentrates for bacterial contamination. Such testing (with sterile sampling performed so as to maintain a closed‐bag system) would be expected to save lives and might allow an extension of platelet storage.
Analysis of HBV infection after blood transfusion in Japan through investigation of a comprehensive donor specimen repository
Authors: Matsumoto Chieko , Tadokoro Kenji , Fujimura Kayoko , Hirakawa Shinichi , Mitsunaga Shigeki , Juji Takeo
Published: March 2003

BACKGROUND: To understand the risk of transfusion‐transmitted viral infection, it is important to precisely assess cases of infection that follow transfusion.

STUDY DESIGN AND METHODS: HBV infections noted after transfusion in 1997, 1998, and 1999 were analyzed. Transfusion in all these cases was performed before NAT was adopted for donor screening. To detect viral infection, PCR and serologic tests for HBV were performed retrospectively on all blood samples from implicated donors that had been stored in a frozen state after each donation. The concentration of HBV genome was measured in HBV‐positive blood samples.

RESULTS: One hundred three cases of HBV infection were analyzed; of these, only 16, including at least 10 infections due to window‐period (HBsAg‐positive by reverse particle hemagglutination assay) donations, were confirmed by further testing to be related to transfusion. The concentrations of HBV genome were very low in four blood samples (<50, 400, 500, and 800 genome equivalents/mL of plasma).

CONCLUSIONS: The remaining risk of transfusion transmission of HBV infection before the adoption of NAT was mainly due to window‐period donations, including one that was made before the HBV genome was detectable by PCR. However, it was determined that transfusion was not responsible in many cases for HBV infection after transfusion.
In vivo survival of apheresis RBCs, frozen with 40‐percent (wt/vol) glycerol, deglycerolized in the ACP 215, and stored at 4°C in AS‐3 for up to 21 days
Authors: Valeri C. Robert , Ragno Gina , Pivacek Linda , O'Neill E. Mary
Published: March 2003

BACKGROUND: The FDA has approved the storage of frozen RBCs at –80°C for 10 years and the postwash storage at 4°C for no more than 24 hours. The 4°C postwash storage period is limited to 24 hours, because the current deglycerolization systems are functionally open systems.

STUDY DESIGN AND METHODS: Two units of RBCs were collected from each of 13 healthy male volunteers. The RBCs were collected in CP2D by the FDA‐approved protocol for an automated apheresis device (MCS, LN8150, Haemonetics) and were stored at 4°C in AS‐3 for 6 days. Using a single disposable glycerolization set in an automated, functionally closed system (ACP 215, Haemonetics) each unit was transferred to a 1000‐mL PVC plastic bag and glycerolized to a concentration of 40‐percent (wt/vol) glycerol and frozen at –80°C. A single disposable deglycerolization set in the ACP 215 was used to deglycerolize the 2 units from the same donor. The deglycerolized RBCs were stored at 4°C in AS‐3 for as long as 21 days.

RESULTS: The mean ± SD freeze‐thaw‐wash recovery value was 89.4 ± 3 percent. The residual hemolysis in the RBCs stored at 4°C in AS‐3 for 21 days after deglycerolization was 0.9 ± 0.2 percent, and the units were negative for both aerobic and anaerobic bacteria. The mean Nageotte WBC count was 9 × 106 per unit. When the deglycerolized RBCs were given as an autologous transfusion after storage at 4°C in AS‐3 for the 7‐ to 18‐day period, the mean ± SD 24‐hour posttransfusion survival was 77 ± 7 percent, and the index of therapeutic effectiveness was 69 ± 8 percent.

CONCLUSION: Two units of human RBCs collected from a single donor by apheresis in the MCS using an LN8150 set can be glycerolized sequentially with a single disposable set and deglycerolized sequentially with another single disposable set in the ACP 215. The previously frozen RBCs stored in AS‐3 for 7 to 18 days at 4°C had acceptable hemolysis and an acceptable mean 24‐hour posttransfusion survival value and index of therapeutic effectiveness.
Influence of cell‐free Hb on local tissue perfusion and oxygenation in acute anemia after isovolemic hemodilution
Authors: Tsai Amy G.
Published: March 2003

BACKGROUND: Oxygen‐carrying solutions are intended to eliminate the blood transfusion trigger. Their ability to maintain microvascular perfusion and to deliver oxygen to tissue when they replace the RBCs as oxygen carriers has not been directly measured.

STUDY DESIGN AND METHODS: Microvascular response to exchange transfusion with a polymerized bovine cell‐free Hb (PBH) solution after acute isovolemic hemodilution with a plasma expander was investigated by using the hamster window model. In vivo functional capillary density (FCD), blood flow, and high‐resolution oxygen distribution in microvascular networks were measured by noninvasive methods.

RESULTS: Exchange transfusion of PBH solution after a 60‐percent isovolemic hemodilution with dextran 70 (MW, 70 kDa) resulted in a Hct of 11 percent and a Hb content of 6.7 g per dL. FCD was 0.37 of baseline. Interstitial pO2 was reduced from 21.0 mm Hg to 0.3 mmHg. Arteriolar and venular blood flows were ratios of 0.75 and 0.76 relative to baseline. In a previous study, tissue pO2 after hemodilution to 5.6 g of Hb per dL with dextran 70 was 23.0 mmHg. Hypervolemic injection of PBH solution increased blood pressure and caused vasoconstriction.

CONCLUSION: Using PBH solution to replace RBC oxygen‐carrying capacity during low Hb content conditions (<50%) causes abnormally low tissue oxygenation and FCD, while the same level of hemodilution with dextran maintains normal microvascular conditions.
First results using automated epifluorescence microscopyto detect Escherichia coli and Staphylococcus epidermidisin WBC‐reduced platelet concentrates
Authors: Seaver Mark , Crookston James C. , Roselle Dominick C. , Wagner Stephen J.
Published: March 2003

BACKGROUND: Many methods have been tested for the detection of bacterial contamination in platelets. However, only those using molecular biology or cell culturing consistently detect contamination at levels below 105 bacteria per mL. This report describes the initial investigation into an alternative method that offers the possibilities of high sensitivity and rapid response while using available laboratory equipment and supplies. This method relies on a fluorescent nucleic acid stain, which preferentially stains bacteria but not platelets, and automated epifluorescence microscopy for rapid analysis. Measurements in WBC‐reduced platelet concentrates (PCs) contaminated with bacteria are reported at concentrations between 103 and 106 bacteria per mL.

STUDY DESIGN AND METHODS: Staphylococcus epidermidis or Escherichia coli was inoculated into aliquots of WBC‐reduced PCs on Days 2 through 5 of storage. Bacterially inoculated and control PCs were stained, platelets and residual WBCs were lysed, and 200 μL of sample was filtered onto black polycarbonate filters. All preparations were done in triplicate. An automated epifluorescence microscope examined approximately 2 percent of the area of each filter and used image analysis to select the fluorescent particles that should be counted as bacteria.

RESULTS: Samples containing 3 to 5 × 103 bacteria per mL produced about three times as many fluorescent particles classified as bacteria as the controls. Lower concentrations of S. epidermidis were detected because of higher fluorescence intensity. Simultaneous preparation of six samples requires about 35 minutes. Analysis of each prepared sample takes 10 minutes, for a total preparation and analysis time of about 95 minutes for 6 samples.

CONCLUSION: Low concentrations (<5 × 103 bacteria/mL) of deliberately inoculated S. epidermidis or E. coli can be measured quickly in WBC‐depleted PCs by using a fluorescent nucleic acid stain, differential lysis, and automated microscopy. Continued refinement of the method, studies employing other bacterial strains, and further validations of assay performance are warranted.
The role of growth factor administration and T‐cell recovery after peripheral blood progenitor cell transplantation in the treatment of solid tumors: results from a randomized comparison of G–CSF and GM–CSF
Authors: Pierelli Luca , Perillo Alessandro , Ferrandina Gabriella , Salerno Giovanna , Rutella Sergio , Fattorossi Andrea , Battaglia Alessandra , Rughetti Aurelia , Nuti Marianna , Cortesi Enrico , Leone Giuseppe , Mancuso Salvatore , Scambia Giovanni
Published: March 2003

BACKGROUND: Peripheral blood progenitor cell (PBPC) transplantation (PBPCT) combined with post‐PBPCT administration of myelopoietic growth factors is a valid therapeutic intervention to rapidly restore hematopoiesis after the delivery of intensive, myeloablative cancer chemotherapy. On the other hand, the best growth factor regimen to potentiate PBPC‐mediated immunohematopoietic recovery has yet to be determined.

STUDY DESIGN AND METHODS: In a randomized evaluation, the effects produced by post‐PBPCT G–CSF and GM–CSF on myeloid/lymphoid recovery and transplant outcome in women with chemosensitive cancer were compared. Thirty‐seven ovarian cancer patients and 34 breast cancer patients ranging in age from 24 to 60 years were treated with carboplatin, etoposide, and melphalan (CEM) high‐dose chemotherapy and then randomly assigned to receive G–CSF (5 μg/kg subcutaneously) or GM–CSF (5 μg/kg subcutaneously) until Day 13 after PBPCT. Patients were compared in regard to hematopoietic recovery, posttransplant clinical management, and immune recovery. Finally, clinical outcome was estimated as time to progression and overall survival.

RESULTS: Hematopoietic recovery and posttransplant clinical management were comparable in both the G–CSF and GM–CSF series. Conversely, significantly higher T‐cell counts were observed in G–CSF‐treated patients during the early and late posttransplant follow‐up. Patients who received G–CSF showed a significantly longer median time to progression. A parallel analysis revealed that patients in whom a higher CD3+ count was recovered had a significantly longer overall survival and time to progression.

CONCLUSION: The enhancement of post‐PBPCT T‐cell recovery observed in G–CSF‐treated patients encourages the use of G–CSF to ameliorate immune recovery, which seems to play a role in post‐PBPCT control of disease in cancer patients. GM–CSF might be administered to prolong immunosuppression after autologous PBPCT for autoimmune diseases or allogeneic PBPCT.
RBC storage for 11 weeks
Authors: Hess John R. , Rugg Neeta , Gormas Jenny K. , Knapp Amy D. , Hill Heather R. , Oliver Cynthia K. , Lippert Lloyd E. , Silberstein Edward B. , Greenwalt Tibor J.
Published: March 2003

BACKGROUND: Increasing the length of RBC storage can increase both RBC availability and quality. This work addresses 11‐week RBC storage in experimental ASs (EASs).

STUDY DESIGN AND METHODS: Three studies were performed. In the first, 24‐hour in vivo recovery of 51Crlabeled autologous RBCs was measured in nine volunteers after storage of their RBCs for 11 weeks in EAS 67. In the second study, 4 units of blood were divided and stored in aliquots with an EAS containing 0, 15, 30, or 45 mmol per L of mannitol; then hemolysis, RBC morphology, and microvesicle protein were measured. In the third study, 6 full units were stored for 12 weeks in the EAS containing 30 mmol per L of mannitol, with weekly sampling for morphologic and biochemical measures of RBC quality.

RESULTS: RBCs stored for 11 weeks in EAS‐67 had a mean 24‐hour in vivo recovery of 79 ± 5 percent, but the hemolysis was 1.35 ± 0.68 percent. Increasing mannitol content of the EAS reduced hemolysis but increased microvesiculation. EAS‐76, with 30 mmol per L of mannitol allowed 11‐week storage with 0.48 ± 0.10 percent hemolysis at 11 weeks and 0.62 ± 0.14 percent hemolysis at 12 weeks.

CONCLUSION: It is possible to store RBCs for 11 weeks in EAS with greater than 75 percent recovery and less than 1 percent hemolysis.
Storage of platelets in additive solutions: effects of magnesium and/or potassium
Authors: De Wildt‐Eggen Janny , Schrijver Johanna G. , Bins Maarten , Gulliksson Hans
Published: March 2003

BACKGROUND: Previous studies indicate that platelet concentrates (PCs) in a platelet additive solution (PAS) containing citrate, acetate, and sodium chloride (PAS‐2) show a significantly higher increase of CD62+ platelets than PCs in other brands of PAS containing Mg2+ and K+. To investigate whether this difference can be explained by the presence of Mg2+ and/or K+ in the storage medium, we performed paired studies comparing storage of PCs in PAS‐2 to PAS‐2 with either Mg2+ or K+ or both in combination.

STUDY DESIGN AND METHODS: PCs from pooled buffy coats were prepared in either PAS‐2 or PAS‐2 with Mg2+ or K+ or both in combination (PAS‐2 modified). Different volumes of MgCl2 solution (1 mol/L) and/or KCl solution (1 mol/L) were added to PAS‐2 to obtain various concentrations. After preparation and during storage (at Days 3 and 7), pH, pCO2, pO2, HCO3−, and CD62 (%) were measured.

RESULTS: During 7 days of storage, pH was very stable (6.9‐7.2) in all PCs. At Day 7, platelet CD62 expression was 49 percent (PAS‐2), 41 percent (PAS‐2 with 1.5 mmol/L Mg2+), and 38 percent (PAS‐2 with 4.5 mmol/L Mg2+). With added K+, at Day 7, expression of CD62 was 55 percent (PAS‐2), 39 percent (PAS‐2 with 4.5 mmol/L K+), and 35 percent (PAS‐2 with 9.0 mmol/L K+). In PAS‐2 modified (PAS‐2 with 1.5 mmol/L Mg2+ and 4.5 mmol/L K+) and CPD plasma, the corresponding CD62 values were 23 and 35 percent, respectively.

CONCLUSION: The combination of Mg2+ and K+ gave significantly (p < 0.05) lower platelet CD62 expression in the storage medium than in PAS‐2. The effects of these differences on platelet metabolism and in vivo properties remain to be investigated.
In utero development of a warm‐reactive autoantibody in a severely jaundiced neonate
Authors: Blackall Douglas P. , Liles Linda H. , Talati Ajay J.
Published: March 2003

BACKGROUND: The fetus and neonate are widely considered to be immunologically immature. However, there are rare case reports of RBC alloantibody and autoantibody development.

STUDY DESIGN AND METHODS: This report describes the case of a severely jaundiced full‐term boy neonate presenting at birth with an IgG warm‐reactive autoantibody.

RESULTS: Mother and neonate were both blood group A, D+. The mother had a negative antibody screen at 18 weeks' gestation and a negative DAT and antibody screen at the time of delivery. The neonate was born with a strongly reactive DAT (IgG) and a panreactive eluate. The serum also contained a panreactive antibody, and all crossmatches were incompatible. The neonate had a bilirubin of 12.5 mg per dL at birth, which peaked at 22.5 mg per dL. However, there was no overt evidence of hemolysis, as evidenced by normal serial Hct levels and reticulocyte counts. The neonate responded well to phototherapy and did not require either simple or exchange transfusion. The neonate's warm‐reactive autoantibody maintained its original strength of reactivity on follow‐up testing performed at 2 weeks and 2 months of age.

CONCLUSIONS: This report describes a rare case of apparent in utero RBC autoantibody development. The fetal/neonatal immune response to blood group antigens is reviewed.
Effectiveness of WBC reduction in neonates: what is the evidence of benefit?
Authors: Fergusson Dean , Hébert Paul C. , Barrington Keith J. , Shapiro Stanley H.
Published: March 2003

BACKGROUND: The presence of WBCs in RBCs is thought to be associated with a number of significant adverse effects in recipients. In adults, WBC reduction has been shown to reduce the frequency of HLA alloimmunization, CMV and HTLV infections, and febrile nonhemolytic transfusion reactions. However, neonates are unique, given that they have an immature immune system and are frequently transfused with RBCs. Thus, the aims of this systematic review were to determine whether WBC reduction of RBCs transfused to neonates decreases the transmission of CMV, reduces the ability to develop HLA antibodies, or reduces the risk of immunomodulation. In addition, nosocomial infection, mortality, and duration of stay were identified and analyzed.

STUDY DESIGN AND METHODS: All studies of WBC reduction were identified by a systematic review of the literature. Studies meeting the inclusion criteria were grouped based on study outcome. Where appropriate, studies were pooled to obtain an overall measure of effect.

RESULTS: Nine eligible studies were identified from the systematic literature search, and six were deemed evaluable. Two studies evaluated WBC reduction and the development of CMV, with different results. The pooled OR was 0.19 (95% CI, 0.01‐3.41), suggesting a clinical but nonsignificant effect. Two studies evaluated WBC reduction and HLA antibody development. As with CMV, the two studies were not congruent in their results. The pooled OR was 0.17 (95% CI, 0.01‐2.43). As for immunomodulation, two small studies presented evidence of a statistically significant change in lymphocyte subsets. No studies were identified with a primary objective of evaluating the impact of WBC reduction on nosocomial infection, mortality, or duration of stay.

CONCLUSION: Current evidence suggests that WBC reduction may be effective in neonates; however, further studies are needed. The lack of convincing data and the significant cost of WBC reduction mandate evaluations to determine the clinical and economic impact.
Reticulated platelets predict platelet count recovery following chemotherapy
Authors: Wang Chao , Smith Brian R. , Ault Kenneth A. , Rinder Henry M.
Published: March 2003

BACKGROUND:A laboratory measure that predicted the timing of platelet recovery after chemotherapy could guide prophylactic platelet transfusion. Reticulated platelets (RPs) are the youngest circulating platelets; an increased percentage of RPs is diagnostic of increased marrow platelet production, such as seen with idiopathic thrombocytopenic purpura, whereas a low percentage of RPs with thrombocytopenia indicates marrow suppression. This study examined whether the percentage of RPs, in combination with a newly devised measurement of "stress thrombopoiesis," the RP maturation index (RP‐MI), could predict platelet count recovery following chemotherapy‐induced thrombocytopenia.

STUDY DESIGN AND METHODS:Platelet count nadirs were retrospectively determined in 35 chemotherapy‐induced thrombocytopenia patients; percentage of RPs and RP‐MI values were assayed at the early nadir (no imminent platelet recovery) and the late nadir (imminent platelet recovery). The latter was defined by a platelet count increase of 20 × 109per L or more in the subsequent 48 hours without platelet transfusion.

RESULTS:Early in the nadir (when platelet recovery did not occur in the subsequent 48 h after sampling), a low RP‐MI and a low percentage of RPs were found in 29 of 35 patients. Late in the nadir, when recovery was imminent, 27 of 30 evaluable patients had elevated percentages of RPs or RP‐MI values; the mean time from sampling to an increase of 20 × 109per L or more was 42 hours. The positive and negative predictive values of this assay were 82 and 91 percent, respectively. Furthermore, when thrombocytopenia was severe (platelet count ≤20 × 109/L), an elevated RP‐MI and/or percentage of RPs correctly predicted imminent platelet count recovery in five of five patients.

CONCLUSION:This noninvasive, rapid, whole‐blood assay of stress thrombopoiesis provides reproducible indices for timing platelet recovery following chemotherapy and the potential to optimize the use of prophylactic platelet transfusions in chemotherapy patients.
Platelet transfusions in children: results of a randomized, prospective, crossover trial of plasma removal and a prospective audit of WBC reduction
Authors: Couban Stephen , Carruthers Julie , Andreou Pantelis , Klama Luba N. , Barr Ronald , Kelton John G. , Heddle Nancy M.
Published: March 2003

BACKGROUND : Febrile nonhemolytic transfusion reactions (FNHTRs) complicate 2 to 37 percent of platelet transfusions in adults, but the incidence of such reactions in children is not known. The effectiveness of plasma reduction after storage and WBC reduction of platelet concentrates before storage was studied in pediatric recipients of platelet transfusions.

STUDY DESIGN AND METHODS : In the first study, a prospective randomized crossover design was used in which patients received either unmodified whole‐blood‐derived or apheresis platelets or platelets from which most of the plasma supernatant had been removed just before transfusion. The second study was a prospective audit of recipients of prestorage WBC‐reduced platelets. Children between 3 months and 17 years of age were eligible for both studies. Patients were assessed for signs and symptoms that are characteristic of a reaction during, immediately after, and 2 hours following transfusion.

RESULTS : There were 226 platelet transfusions administered to 66 children. One hundred and sixty transfusions were given to 35 children enrolled in the randomized study, and 66 transfusions were given to 33 children during the audit. In the randomized study, nine of the 75 transfusions of unmodified platelets (12%) and six of 85 transfusions of poststorage plasma‐removed platelets (7%) were associated with an FNHTR (p = 0.42). In the audit, three of 66 transfusions of prestorage WBC‐reduced platelets (5%) were associated with an FNHTR. Allergic reactions occurred with 5 percent (4 of 75), 6 percent (5 of 85), and 6 percent (4 of 66) of platelet transfusions, respectively.

CONCLUSION : FNHTRs appear to be less common among pediatric recipients of platelet transfusions than in adults. In our two studies, there was a trend toward a lower frequency of FNHTRs with poststorage plasma removal and prestorage WBC reduction than with standard platelets, but this was not significant.
Anaphylactic transfusion reactions in haptoglobin‐deficient patients with IgE and IgG haptoglobin antibodies
Authors: Shimada Eiko , Tadokoro Kenji , Watanabe Yoshihisa , Ikeda Kazuyo , Niihara Hiromi , Maeda Ikiko , Isa Kazumi , Moriya Susumu , Ashida Takashi , Mitsunaga Shigeki , Nakajima Kazunori , Juji Takeo
Published: March 2003

BACKGROUND : Patients with haptoglobin deficiency associated with haptoglobin IgG antibodies, who experienced severe nonhemolytic transfusion reactions (NHTRs), have been identified in Japan. Haptoglobin deficiency therefore might be a risk factor for NHTRs.

STUDY DESIGN AND METHODS : A total of 4138 cases of voluntarily reported NHTRs in Japan, including 367 cases of immediate‐onset anaphylactic NHTRs, were examined to identify haptoglobin deficiency. Serum haptoglobin IgG and IgE antibodies were determined in haptoglobin‐deficient patients to elucidate the mechanism underlying the transfusion reactions.

RESULTS
: Seven patients with haptoglobin deficiency were identified. Six of them experienced severe and acute NHTRs. Six of them were identified to be homozygous for the
Hp

del
allele of the haptoglobin gene. Both haptoglobin IgG and IgE antibodies were detected in serum samples of all the patients. The stimulative effects of blood transfusion on the production of hap‐ toglobin antibodies in the patients and the relation‐ ship between the presence of the antibodies and the occurrence of the transfusion reactions were observed.

CONCLUSION : Anaphylactic NHTRs in these patients with haptoglobin deficiency associated with serum haptoglobin antibodies were suggested to be prevalent in Japan. In addition to IgG antibodies, IgE haptoglobin antibodies detected in the sera of such patients were suggested to play a role in the occurrence of the reactions.
The contribution of sociodemographic, medical, and attitudinal factors to blood donation among the general public
Authors: Boulware L.E. , Ratner L.E. , Ness P.M. , Cooper L.A. , Campbell‐Lee S. , LaVeist T.A. , Powe N.R.
Published: March 2003

BACKGROUND : Few studies have simultaneously assessed the relative importance of sociodemographic, medical, and attitudinal factors in explaining which individuals are more likely to donate blood.

STUDY DESIGN AND METHODS : A cross‐sectional telephone survey of households in Maryland was conducted to identify the relation of sociodemographic, medical, and attitudinal factors to blood donation history among the general public. Random digit dialing was used to identify households; individuals aged 18 to 75 years were randomly selected within households. In multivariate analyses, the independent relationship of these factors with prior history of blood donation was assessed, and the amount of variation in prior history of blood donation among the study population that could be explained by these factors was determined.

RESULTS : Of 385 participants (84% of randomized homes), 228 (59%) had donated blood at least once in the past. After adjusting for potential confounders, women, black participants, and those agreeing with the statement “I am afraid of hospitals” had 60 to 80 percent lower odds of prior donation when compared with men, white participants, and those who did not agree with the statement (OR [95% CI]: 0.2 [0.1‐0.4], 0.4 [0.2‐0.8], and 0.3 [0.2‐0.6], respectively). The effect of fear of hospitals was consistent across sex and race. Trust, fear, and suspicion of hospitals were among factors contributing most to variation in prior donation history.

CONCLUSION : Female sex, black race, and fear of hospitals are three major factors negatively associated with prior history of blood donation. Fear of hospitals affects blood donation patterns across race and sex groups. Future study is needed to determine whether recruitment of blood donors may be more efficient if focused toward women, minorities, and donors' fears of healthcare facilities or hospitals.