bluuebunny/crossref_metadata_2025 huggingface dataset:
Allogeneic transplantation combining mobilized blood and bone marrow in patients with refractory hematologic malignancies
Authors: Nemunaitis J. , Rosenfeld C. , Collins R. , Pallansch P. , Pineiro L. , Ohr S. , Waxman D. , Stone M. , Fay J. , Miller W. , Piñeiro Luis
Published: March 2003

BACKGROUND: Mobilized blood stem cells have been used successfully in autologous transplant recipients to reduce the complications of pancytopenia due to dose‐intensive chemotherapy. Reports of cytokine‐ mobilized blood progenitor cells in allogeneic transplant recipients are rare.

STUDY DESIGN AND METHODS: This is a pilot trial of six patients. Patients with advanced hematologic malignancy received bone marrow (median total 2.6 × 10(8) mononuclear cells/kg) followed by four daily transfusions of blood (median total 9.5 × 10(8) mononuclear cells/kg) from HLA‐matched sibling donors who were mobilized with recombinant human granulocyte‐colony‐stimulating factor (5 micrograms/kg/day subcutaneously for 5 days). All patients received cyclosporine and prednisone for graft‐versus‐host disease (GVHD) prophylaxis.

RESULTS: An absolute neutrophil count greater than 500 per mm3 was achieved on Day 12, and platelet transfusion independence was achieved on Day 16. The median day of hospital discharge was Day 23 after transplant. All patients achieved 100‐percent donor cell engraftment. Acute > or = Grade III GVHD did not develop in any patients, but all patients developed Grade I (n = 4) or Grade II (n = 2) acute GVHD. Chronic extensive GVHD developed in four of six patients. One patient died of pneumonia 263 days after transplant while undergoing immune‐suppressive therapy for chronic GVHD.

CONCLUSION: The transfusion of blood stem cells in patients undergoing allogeneic bone marrow transplant is well tolerated soon after transplant, but the development of chronic GVHD may limit the general usage of unmanipulated blood stem cells.
Prestorage white cell reduction in saline‐adenine‐glucose‐mannitol red cells by use of an integral filter: evaluation of storage values and invivo recovery
Authors: Rogers S.E. , Edmondson D. , Goodrick M.J. , Standen G.R. , Franck V. , Reppucci A. , Pamphilon D.H.
Published: March 2003

BACKGROUND: Prestorage white cell (WBC) reduction in blood components may decrease the incidence of adverse reactions and improve component quality. A bottom‐and‐top system with an integral third‐generation WBC‐ reduction filter has been studied.

STUDY DESIGN AND METHODS: Whole blood was collected from 30 healthy donors: from 20 by using a blood container system with an integral filter and from 10 controls by using a standard blood container system. Ten test units were buffy coat‐ depleted, stored for 72 hours at 4 degrees C, and then filtered, while an additional 10 test units were buffy coat‐depleted and filtered at room temperature within 8 hours of collection. All units were stored at 4 degrees C for 42 days and sampled weekly.

RESULTS: The mean WBC content of the 72‐hour, 4 degrees C units was 0.33 × 10(6), that of the room‐temperature units was 2.6 × 10(6), and that of the buffy coat‐ depleted controls was 460 × 10(6) (p < 0.0005). No significant differences were found among lactate, glucose, sodium, potassium, and plasma hemoglobin levels in the three groups. ATP and 2,3 DPG levels were significantly better preserved in control units than in 72‐hour, 4 degrees C units (p = 0.016 and p = 0.032, respectively), but not better than in the room‐temperature units. Significant differences were observed between pH values in filtered units and both groups of test units (p = 0.016). In biologic terms however, these differences were small. Red cells from an additional eight healthy volunteer donors were processed by an 8‐hour room‐temperature method and stored for 35 days. Studies in vivo 24‐hour recovery of autologous red cells were performed by transfusing a radiolabeled (51Cr plus 131I‐albumin) aliquot after 35 days' storage. Good recovery (mean > 80%) was found by both the single‐ and double‐isotope‐label methods. Recovery was significantly greater when calculated by the single‐isotope method (p = 0.02).

CONCLUSION: The combination of buffy coat removal and filtration in the blood container system with an integral filter achieved effective WBC reduction (> or = 3 log10 reduction from whole blood) without biologically significant detriment to in vitro or in vivo storage values.
Suction‐induced hemolysis at various vacuum pressures: implications for intraoperative blood salvage
Authors: Gregoretti S.
Published: March 2003

Background: Suction vacuum pressure (VP) not exceeding 150 torr is recommended during intraoperative blood salvage to minimize hemolysis. When the suction provided by this VP becomes inadequate because of brisk bleeding, an alternative high‐VP suction should be used and the blood discarded. This approach often results in the salvage of only a small fraction of the blood shed during surgery complicated by large hemorrhage.

Study Design and Methods: This laboratory study was designed to quantitate the hemolysis caused by various VPs in a suction system commonly used for intraoperative blood salvage. For each experiment, a batch of blood with a hematocrit of 30 to 35 percent was prepared by mixing of outdated units of red cells, fresh‐frozen plasma, and saline solution. Aliquots of this blood were suctioned at VPs of 150, 200, 250, and 300 torr, either without (6 experiments) or with (4 experiments) maximal air entrainment. Total hemoglobin, hematocrit, red cell count, plasma‐free hemoglobin, and serum potassium were measured in the blood before suction and in each aliquot after suction.

Results: Suction of blood mixed with air caused much greater hemolysis than suction of blood alone (p < 0.01 at each VP tested). Raising the VP from 150 to 300 torr increased hemolysis from 0.14 +/− 0.20 percent (mean +/− SD) to 0.32 +/− 0.21 percent (p < 0.05) when blood alone was aspirated and from 1.45 +/− 0.50 percent to 2.85 +/− 0.22 percent (p < 0.05) when blood was suctioned with air. With either type of suction, red cell count, hematocrit, and serum potassium did not change significantly throughout the range of VPs tested.

Conclusion: Hemolysis was found to depend on the VP applied and, to a much greater extent, on the amount of blood and air mixing. Increasing the VP above the recommended limit of 150 torr was not associated with inordinate hemolysis. Even when a VP as high as 300 torr was used, hemolysis ranged between 0.3 and 3.0 percent, depending on whether air was suctioned with the blood or not. The data support the idea that the lowest VP compatible with a clear surgical field should be used during intraoperative blood salvage and that the suctioning of air should be avoided as much as possible. These data also suggest that, in contrast to current recommendations, suction VP during intraoperative blood salvage can be increased up to 300 torr if required by the rate of bleeding, without causing excessive hemolysis.
Photochemical inactivation of duck hepatitis B virus in human platelet concentrates: A model of surrogate human hepatitis B virus infectivity
Authors: Eble B. E. , Corash L.
Published: March 2003

BACKGROUND: Photochemical decontamination of platelet concentrates (PCs) has been demonstrated by the use of 8‐methoxypsoralen and ultraviolet A light. Systems for studying the inactivation of blood‐ borne viruses facilitate the evaluation of photochemical decontamination protocols.

STUDY DESIGN AND METHODS: Duck hepatitis B virus (HBV), a model for human HBV, was adapted for the study of hepadnavirus inactivation. A highly specific in vitro infectivity assay used primary duck hepatocyte cultures and was followed by the detection of replicated duck HBV sequences. RESULTS: Duck HBV‐infected primary duck hepatocyte cultures produced authentic infectious virus. High‐ titer (> 10(9) virus genome equivalents/mL) duck HBV‐infected sera were completely inactivated in serum or PCs by the use of 100 micrograms per mL of 8‐methoxypsoralen and 70 J per cm2 of ultraviolet A light. Intracellular duck HBV (> 4.2 log10) in PCs was also inactivated. Culture results were confirmed by a sensitive duckling infectivity assay that indicated that 6.3 log10 of infectious duck HBV had been inactivated by photochemical decontamination.

CONCLUSION: The sensitivity of the culture assay was comparable to that of the duckling assay using polymerase chain reaction gene amplification to detect duck HBV. Duck HBV inactivation in PCs was dependent on the dose of ultraviolet A light and independent of 8‐methoxypsoralen concentrations of 100 to 300 micrograms per mL: 100 micrograms per mL 8‐ methoxypsoralen inactivated 4 to 5 log10 of virus in conjunction with 20 to 40 J per cm2 of ultraviolet A light. The polymerase chain reaction‐enhanced duck HBV culture system has utility in optimizing photochemical decontamination protocols.
Acute gram‐negative urosepsis mimicking an acute hemolytic transfusion reaction
Authors: Friedmany H. D. , Noonan M.
Published: March 2003

BACKGROUND: The acute hemolytic transfusion reaction (AHTR) is one of the most feared complications of blood transfusion. Over the years, several clinical conditions, as well as errors in blood component preparation and administration, that mimic AHTR have been identified. This report describes a novel variation on the theme of pseudo‐AHTR.

CASE REPORT: A 47‐year‐old diabetic man with drug‐induced pancytopenia suddenly manifested severe shaking chills, flank pain, and back pain during a red cell transfusion. The passage of bright red urine immediately after the transfusion virtually confirmed for the clinicians administering the transfusion that an AHTR had occurred. In the laboratory, the hematuria was shown to be due principally to red cells and not to free hemoglobin. Further posttransfusion work‐up showed a urinary tract infection and overwhelming bacterial sepsis with Escherichia coli. CONCLUSION: As a pseudo‐AHTR, gram‐negative bacterial sepsis of urinary tract origin may surpass other forms of sepsis. Urosepsis should be considered in the work‐up of a suspected AHTR in a pancytopenic patient with a urinary tract infection.
Hepatitis C virus (HCV) genotype analysis in apparently healthy anti‐ HCV‐positive Parisian blood donors
Authors: Stuyver L. , Fretz C. , Esquivel C. A. , Boudifa A. , Jaulmes D. , Azar N. , Lunel F. , Leroux‐Roels G. , Maertens G. , Fournel J. J.
Published: March 2003

BACKGROUND: As only a few studies have examined the prevalence of various hepatitis C virus (HCV) subtypes in blood donors, information about the variability and route of infection in apparently healthy persons is limited. STUDY DESIGN AND METHODS: Blood donations collected at a large Parisian hospital (52,441) were investigated for antibodies to HCV. Serum samples were screened with an enzyme immunoassay. All HCV‐ positive donations were retested with a second enzyme immunoassay and confirmed by immunoblot. The HCV genotype was determined for all polymerase chain reaction‐positive subjects. Untypable genotypes were sequenced in the NS5B region. RESULTS: In total, 83 (0.26%) blood donors were anti‐HCV positive. Men (0.34%) were significantly more likely to be infected (p < 0.001) than women (0.19%). Prevalence rates in men between 20 and 39 years of age were higher than those in similar women (p = 0.01), but greater in women aged from 50 to 65 years (p = 0.05). Fifty‐five sera were viremic, of which 49 could be genotyped by a line probe assay. One new HCV type 1 subtype and three new HCV type 2 subtypes were discovered. In total, 28, 10, 11, 5, and 1 serum samples were grouped into HCV types 1 through 5, respectively, involving a total of 13 subtypes. The mean age of HCV type 2‐infected donors was 42 +/− 11 years, but that for type 3‐infected subjects was only 30 +/− 4 years (p = 0.0048). Forty‐nine subjects showed elevated alanine aminotransferase levels; 39 (80%) of these subjects were viremic (p < 0.05). CONCLUSION: Among the sampled population, an HCV prevalence rate of 0.26 percent was found, with the five most common European genotypes causing the infections. Four new subtypes were discovered. Correlation between genotype and risk factors was not apparent, but links with age, sex, and ethnic origin emerged.
A survey of red cell use in 45 hospitals in central Ontario, Canada
Authors: Chiavetta J.A. , Herst R. , Freedman J. , Axcell T. J. , Wall A. J. , van Rooy S.C.
Published: March 2003

BACKGROUND: The purpose of this survey was to establish baseline information on blood component use in relation to patient diagnoses, procedures, and demographics and to identify patterns of blood use that may be used for blood program planning and transfusion audits. STUDY DESIGN AND METHODS: A cross‐sectional survey of the transfusion of blood components in teaching and nonteaching hospitals in central Ontario between September 1991 and August 1992 was carried out. Coders of hospital medical records routinely record demographics, procedures, diagnoses, and other relevant information. A protocol was created by which medical records coders could add the components transfused to the discharge abstract for this study. Red cell use is reported here. RESULTS: Of the 61 hospitals invited to participate, from which 547,279 patients were discharged during the 12‐month period of the study, 45 (74%) agreed to participate. Information was collected on 439,373 discharged patients. Of these, 26,611 (6.1%) received at least 1 unit of red cells. Of a total of 101,116 red cell units transfused, more than 74 percent were used in patients discharged with neoplasms, gastrointestinal diseases, circulatory system diseases, and trauma. High‐transfusion‐use procedures included operations and procedures on the digestive and cardiovascular systems, diagnostic and therapeutic procedures, musculoskeletal system, and hemic or lymphatic system procedures. CONCLUSION: This survey provides baseline blood transfusion information for a specific period that can help determine the need for hospital audits and maximum surgical blood‐order schedule guideline reviews. This information is relevant to current recommendations to reduce patient's exposure to blood components. These transfusion data will assist in blood program planning based on known disease trends, demographics, and population changes.
Removal of soluble biologic response modifiers (complement and chemokines) by a bedside white cell‐reduction filter
Authors: Snyder E. L. , Mechanic S. , Baril L. L. , Davenport R.
Published: March 2003

BACKGROUND: Biologic response modifiers infused with stored platelet concentrates (PCs) are believed to contribute to symptoms seen during transfusion reactions. Although prestorage white cell reduction is known to decrease the production of some biologic response modifiers during storage, the possibility that poststorage (bedside) white cell reduction could reduce the amount of biologic response modifiers already present in stored PCs during bedside filtration has not been well studied. STUDY DESIGN AND METHODS: Individual PCs were pooled on storage Days 2 and 5 and passed through a third‐generation white cell‐ reduction filter. The results from a series of in vitro PC assays were studied, before and immediately after filtration, as were levels of C3a and interleukin 8 (n = 5). Levels of other biologic response modifiers‐ C5a, interleukin 1 beta, interleukin 6, tumor necrosis factor alpha, and RANTES‐were also studied. Removal of interleukin 8 and RANTES was studied further by using serial filtration of units of PC. RESULTS: For the in vitro platelet assays studied, pH was unchanged after filtration from prefiltration values in units of PCs pooled on storage Day 2 or 5. A 4 log10 reduction in white cells was reliably seen after filtration in Day 2 and 5 pooled PCs. Postfiltration platelet loss was 14.8 percent for Day 2 pooled PCs and 9.6 percent for Day 5 pooled PCs. For pools of both Day 2 and Day 5 platelets, postfiltration levels of CD62 (P‐selectin, CD62P) were unchanged from prefiltration levels, as were results for morphology scores. Levels of C3a decreased after filtration in both the Day 2 pooled PCs (448 ng/mL before filtration vs. 20 ng/mL after filtration) and the Day 5 pooled PCs (1976 ng/mL before filtration vs. 124 ng/mL after filtration). Levels of interleukin 8 were similarly reduced after filtration in the Day 2 pooled platelets (188 pg/mL before filtration vs. 27 pg/mL after filtration) and the Day 5 pooled platelets (2234 pg/mL before filtration vs. 799 pg/mL after filtration). Levels of interleukin 8 in other components evaluated after filtration declined similarly. However, levels of the proinflammatory cytokines interleukin 1 beta and interleukin 6 did not decline after filtration. Serial filtration studies showed that, although levels of interleukin 8 and RANTES were initially lowered by filtration, they returned to prefiltration values with increases in the volume of filtration. CONCLUSION: The third‐generation bedside filter used in this study reliably reduced the level of white cell contamination to 4 log10 white cells per PC. It also lowered the levels of interleukin 8, RANTES, and C3a. The filter did not, however, remove (scavenge) the proinflammatory cytokines interleukin 1 beta and 6. The mechanism of chemokine and C3a removal by the filter is unknown, but it may be related to ionic interactions between these biologic response modifiers and the filter medium.
Considerations of pool size in the manufacture of plasma derivatives
Authors: Lynch T.J. , Weinstein M.J. , Tankersley D.L. , Fratantoni J.C. , Finlayson J.S.
Published: March 2003

Background: The pooling of human plasma from many donors for the purpose of manufacturing therapeutic proteins increases the risk of exposing recipients of these proteins to pathogens that may contaminate 1 or a few units included in the pool.

Study Design and Methods: This risk is estimated for a range of manufacturing scales that would derive material from a varied number of donors and for a number of hypothetical infectious agents that may exist in the donor population over a wide range of prevalence. Risk is also calculated both for recipients of single doses of a plasma protein and for those who depend on long‐term treatment with plasma derivatives.

Results: Risk of exposure increases with pool size and the prevalence of the agent in question and accumulates with repeated treatments with material manufactured from different pools.

Conclusion: Reducing pool size would at best decrease this risk in proportion to the reduction in manufacturing scale. However, for individuals requiring repeated or continuous treatments, the risk of exposure to all but the rarest infectious agents would be only minimally affected, even by large reductions in manufacturing scale.
A survey of blood component use in a German university hospital
Authors: Zimmermann R. , Buscher M. , Linhardt C. , Handtrack D. , Zingsem J. , Weisbach V. , Eckstein R.
Published: March 2003

BACKGROUND: There are no recent studies on transfusion practice and blood use with regard to diagnoses of European recipients. We conducted a survey of blood component use, including packed red cells, fresh‐ frozen plasma, and platelets, in an acute‐care university hospital in the Greater Nurnberg area. STUDY DESIGN AND METHODS: A survey was carried out of blood component transfusion at a university hospital (Erlangen, Germany) between June 1994 and May 1996. Transfused units were listed by broad diagnostic categories formed from principal diagnoses of the recipients according to the International Classification of Diseases, Ninth Revision. RESULTS: Among 100,497 discharged patients, 6,590 patients who received transfusion (6.6%) are represented in this survey. Of 28,440 red cell units and 8,592 fresh‐ frozen plasma units, 72.4 percent and 66.9 percent, respectively, were used in patients with neoplastic diseases, circulatory system diseases, or disorders of the digestive system. Of 2704 platelet units, 78.1 percent were transfused to patients with neoplastic or gastrointestinal diseases or diseases of blood‐forming organs. These four diagnostic categories accounted for 77.7 percent of all costs of transfusion therapy. Males received 60.1 percent of all blood components transfused, and patients less than 65 years old received 68.0 percent. CONCLUSION: This survey provides information on blood component usage in a German university hospital. It demonstrates the concentration of today's blood utilization among a few diagnostic categories. The study shows that detailed information on local blood use may be obtained quickly by using data available from transfusion services and medical record departments. This information is relevant for quality management of transfusion practice, cost analyses and for planning local and regional blood donation programs.
Does a patient with hereditary spherocytosis qualify for preoperative autologous blood donation?
Authors: Weinstein R. , Martinez R. , Hassoun H. , Palek J.
Published: March 2003

BACKGROUND: Hereditary spherocytosis (HS) is characterized by osmotically fragile spherocytic red cells (RBCs), mild to moderate hemolysis, and splenomegaly. Little is known about the safety of blood bank storage of HS RBCs. CASE REPORT: A 50‐year‐old man with mild autosomally dominant HS asked to make an autologous RBC deposit before his scheduled surgery. His RBCs were serially tested for osmotic fragility during 4 weeks of blood bank storage in CPD with adenine‐ saline. Aliquots of his fresh and 4‐week‐old RBCs were also labeled with 51Cr for measurement of in vivo survival. The osmotic fragility of the patient's fresh RBCs was normal, with 50‐percent hemolysis at approximately 0.43‐percent NaCl and a sigmoid lysis curve. Incubated osmotic fragility (24 hours at 37 degrees C) was abnormal, with 50‐ percent hemolysis at 0.64‐percent NaCl. Median survival of his fresh 51Cr‐labeled RBCs was moderately shortened at 17.5 days. Osmotic fragility increased during storage, with 50‐percent lysis occurring at 0.58‐percent and 0.62‐percent NaCl after 2 and 4 weeks, respectively. Volunteer normal donor RBCs exhibited 50‐percent lysis in 0.48‐percent NaCl after 4 weeks. Median survival of the patient's 4‐week‐old 51Cr‐ labeled RBCs was severely shortened, at 3.5 days. At surgery, intraoperatively salvaged RBCs demonstrated osmotic fragility identical to that of the patient's RBCs freshly obtained by venipuncture. CONCLUSION: HS RBCs may lose membrane under optimum storage conditions, becoming unsuitable for transfusion at surgery. Intraoperative autologous transfusion may be preferable to preoperative deposit for avoidance of allogeneic RBC transfusion in patients with HS.
Restricted use of T‐cell receptor V beta genes in posttransfusion graft‐ versus‐host disease
Authors: Wang L. , Tadokoro K. , Tokunaga K. , Uchida S. , Moriyama S. , Bannai M. , Mitsunaga S. , Takai K. , Juji T.
Published: March 2003

BACKGROUND: Posttransfusion graft‐versus‐host disease (PT‐GVHD) is a pathophysiological process of the immune system. Mature donor T cells that have survived the host defense mechanisms recognize recipient allo‐ antigens through antigen‐specific T‐cell receptors and mount an immune attack against certain host antigens. STUDY DESIGN AND METHODS: To characterize the T cells involved in PT‐GVHD, expression of the T‐cell receptor V beta genes in peripheral blood T cells isolated from four PT‐ GVHD patients was analyzed. T‐cell receptor beta chain cDNA derived from mRNA extracted from peripheral blood lymphocytes was amplified by an inverse polymerase chain reaction method, and the cDNA was subsequently cloned and sequenced. RESULTS: The cDNA clones derived from patients with PT‐GVHD showed remarkable differences in the distribution of T‐cell receptor V beta gene use from that in normal controls. Very restricted T‐cell receptor V beta gene repertoires and preferential expression of certain V beta gene segments were revealed in the peripheral blood lymphocytes of each patients. Shared V beta gene use among the patients was not observed, probably because of HLA differences among the patient‐donor combinations. CONCLUSION: This restricted use reflected in vivo expansion of a limited number of engrafted T‐cell clones at the onset of PT‐GVHD and suggested that those oligoclonal donor T cells may be involved in the induction of PT‐ GVHD.
Induction of a secondary human anti‐HLA alloimmune response in severe combined immunodeficient mice engrafted with human lymphocytes
Authors: Lazarus A.H. , Crow A.R. , Semple J.W. , Cosgrave D. , Kalovsky E.J. , Hannach B. , Blanchette V. , Freedman J.
Published: March 2003

BACKGROUND: Experimental manipulation of transfusion‐induced alloimmunization is limited in humans by ethical considerations. Conversely, studies of alloimmunization in animal models may not reflect the human immune system closely enough to be of optimal benefit. The development of an in vivo model of human alloimmunization that is amenable to experimental manipulation is thus desirable. STUDY DESIGN AND METHODS: An in vivo model of human alloimmunization was evaluated by using mice with severe combined immunodeficiency (SCID). SCID mice underwent gamma‐radiation (200 cGy) and received an intraperitoneal injection of human peripheral blood lymphocytes (PBLs) from donors immunized to HLA antigens by prior pregnancy (reconstitution). These Hu [human]‐PBL‐SCID mice were then challenged with HLA‐mismatched PBLs. Alloantibodies were evaluated by flow cytometry and a standard two‐stage microlymphocytotoxicity assay. RESULTS: Hu‐PBL‐SCID mice (n = 22) that were challenged with PBLs expressing the HLA antigens to which the donors had previously been immunized, made significantly more IgM and IgG alloantibodies than did the unchallenged mice. Responses were measurable by 1 week after reconstitution and challenge. Prior treatment of SCID mice with anti‐ asialo GM1, which depletes murine natural killer cells and macrophages, further increased the alloantibody response of challenged mice. The human alloantibodies generated were specific to the challenge HLA antigens as assessed by microlymphocytotoxicity assay. CONCLUSION: Hu‐ PBL‐SCID mice are a useful model system in which to study and manipulate the induction of secondary human alloimmune responses against cellular HLA class I antigens. This model will be valuable for testing the in vivo effect of novel immunotherapies on the inhibition of the human alloantibody response.
The effect of hypotonicity, glutamine, and glycine on red cell preservation
Authors: Greenwalt T.J. , Rugg N. , Dumaswala U.J.
Published: March 2003

BACKGROUND: Red cells (RBCs) stored in hypo‐os‐molar additive solutions with the same concentrations of adenine, dextrose, mannitol, and sodium chloride and varied amounts of ammonium, phosphate, glycerol, and glutamine were better preserved than RBCs in the standard additive solution (Adsol). Cell swelling occurred in all the experimental additives. This observation prompted the evaluation of glutamine and glycine alone, as well as a combination of glutamine and glycine, all of which have been described as producing swelling of rat liver cells.

STUDY DESIGN AND METHODS: Aliquots of RBCs were stored at 4°C in Adsol or experimental additive solutions (EASs) all containing adenine, 2 mM; dextrose, 110 mM; mannitol, 55 mM; and sodium chloride, 50 mM. EAS 42 had, in addition, glutamine, 10 mM; glycine 5 mM; and phosphate, 20 mM. EAS 43 had glutamine, 10 mM; glycine, 10 mM; and phosphate 20 mM. EAS 44 had glutamine, 10 mM; EAS 45 had glutamine, 10 mM, and phosphate, 20 mM; and EAS 46 had only glycine, 10 mM. At intervals, measurements were made of mean corpuscular volume, mean corpuscular hemoglobin concentration, morphology, ATP, hemolysis, supernatant potassium, ammonia, pH, and microvesicles shed.

RESULTS: The initial mean corpuscular volumes were larger in all EASs than in Adsol, but the greatest difference was between EASs 44 and 46 (108 fL) and Adsol (86 fL) (p<0.001). The morphology scores were significantly better in all the EASs (p<0.04). The ATPs were significantly greater in all the EASs (p<0.001), and highest in those with phosphate. Potassium leakage and hemolysis were less in the EASs (p<0.001). The ammonia levels were higher in all the EASs than in Adsol, with the exception of EAS 46. During storage, the extracorpuscular and intracorpuscular pH levels were essentially identical. The shedding of microvesicles was greatly reduced in all the EASs.

CONCLUSION: Cell swelling induced in RBCs after collection appears to improve preservation. Ammonia and phosphate enhance RBC ATP maintenance. Glycine decreases the formation of ammonia by RBCs stored in a hypotonic medium.
Microdroplet fluorochromatic assay for the enumeration of white cells (WBCs) in WBC‐reduced blood components: validation and application for evaluating newly developed WBC‐reduction filters
Authors: Borzini P , Dumont L.J.
Published: March 2003

BACKGROUND: Sensitive and accurate counting methods are required to assess the residual white cells (WBCs) in WBC‐reduced blood components. The Nageotte hemocytometer, widely used for this purpose, is cumbersome, and its efficacy is dependent upon the skill of the operator. The performance of a simple fluorochromatic assay using tissue‐typing microdroplet trays is presented here. STUDY DESIGN AND METHODS: Undiluted samples of blood components were mixed with a fluorochromatic dye in trays. WBCs were counted under an epifluorescence microscope. The accuracy and sensitivity of this method were compared with those of the reference Nageotte hemocytometer method by using serial dilution of samples of platelets and red cells containing known concentrations of WBCs and by calculating the standard curves. The Nageotte hemocytometer and the microdroplet fluorochromatic assay (MFA) were also compared in terms of count correlation and reproducibility in 320 paired counts of plateletpheresis samples. MFA was used to evaluate a newly developed WBC‐reduction red cell filter. RESULTS: The MFA for platelets and red cells was linear to 0.1 and 0.03 WBCs per microL, respectively. The linear regression line of log10 MFA versus log10 Nageotte method had a slope of 0.963, intercept of ‐0.04, and r2 of 0.968. The Nageotte method gave an estimation of WBC content 12 to 20 percent greater than that of the MFA. The MFA, with a larger neat sample volume, showed precision comparable to that of the Nageotte method. The filters demonstrated a median WBC reduction of 4.8 log10. CONCLUSION: The MFA is a sensitive and accurate method for quality control processes to assess the residual WBCs in WBC‐reduced blood components.
A method for estimating hepatitis B virus incidence rates in volunteer blood donors. National Heart, Lung, and Blood Institute Retrovirus Epidemiology Donor Study
Authors: Korelitz J.J. , Busch M.P. , Kleinman S.H. , Williams A.E. , Gilcher R.O. , Ownby H.E. , Schreiber G.B. ,   National Heart, Lung, and Blood Institute Retrovirus Epidemiology Donor Study
Published: March 2003

BACKGROUND: Calculations of the incidence of hepatitis B virus (HBV) infections in the blood donor setting that are based solely on data for seroconversion to hepatitis B surface antigen (HBsAg) will underestimate the incidence due to the transient nature of antigenemia. Estimates based on antibody to hepatitis B core antigen will overestimate the incidence due to false‐positive results caused by the nonspecificity of the test. STUDY DESIGN AND METHODS: Serologic test results were obtained from multiple‐time volunteer donors at five United States blood centers from January 1991 through December 1993. The observed HBsAg seroconversion rate was multiplied by an adjustment factor, derived from the weighted average probability of a positive HBsAg test for HBV‐infected donors who become chronic carriers, for donors with a primary antibody response without detectable antigenemia, and for donors who develop transient antigenemia. RESULTS: Among 586,507 multiple‐time donors giving 2,318,356 donations and observed for 822,426 person‐years, the HBsAg incidence rate was 4.01 per 100,000 person‐years. On the basis of prior reports of the duration of HBsAg positivity and the observed distribution of interdonation intervals among the study group, there was an estimated 53‐percent chance that an HBV‐infected donor with transient antigenemia would have a positive HBsAg test result. If 70 percent of newly HBV‐infected adults have transient antigenemia, 25 percent have a primary antibody response without primary antigenemia, and 5 percent become chronic carriers, the overall chance of being detected by the HBsAg test was 42 percent, for an adjustment factor of 2.38. The total HBV incidence rate, therefore, was estimated to be 9.54 per 100,000 person‐years. CONCLUSION: The crude HBV incidence rate observed from HBsAg test results will underestimate the true rate. The adjusted HBV incidence rate should be used in applications such as estimations of residual HBV risk to the blood supply and projections of the benefits of screening for HBV DNA.
Human immunodeficiency virus (HIV) antibodies detected by new assays that are enhanced for HIV‐1 subtype O
Authors: Jongeriüs John M. , van der Poel Cees L. , Van Loon Anton M. , van den Akker Ruud , Schaasberg Wim , Van Leeuwen Eleonore F.
Published: March 2003

BACKGROUND: Human immunodeficiency virus type 1 (HIV‐1) subtype O infections are not reliably detected by commonly used anti‐HIV‐1/2 screening assays. Therefore, anti‐HIV‐1/2 assays have been modified to increase their sensitivity in detecting antibodies to HIV‐1 subtype O. STUDY DESIGN AND METHODS: Two new anti‐HIV‐1/2 enzyme‐linked immunosorbent assays (ELISAs) (Abbott Plus and Ortho Enhanced) were compared with a currently used anti‐HIV‐1/2 ELISA (Abbott Recombinant) in various serum panels: 91 Western blot‐confirmed anti‐HIV‐1‐positive samples, 20 samples from Western blot‐confirmed HIV‐1‐infected patients in log3 serial dilutions, and 1463 samples from consecutive, volunteer, nonremunerated blood donors. RESULTS: Among 91 anti‐HIV‐1 Western blot‐ positive samples, 2 (2.2%) were missed by the Abbott Recombinant ELISA, but all 91 were detected by the Abbott Plus and Ortho Enhanced ELISAs. In contrast, two discrepant samples were found to react in viral lysate‐ based assays. In serial dilutions, Ortho Enhanced ELISA was significantly less sensitive than the Abbott Recombinant and Abbott Plus ELISAs, with the latter two being of comparable sensitivity. The specificities of Abbott Recombinant, Abbott Plus, and Ortho Enhanced ELISAs in 1463 blood donors were 100, 99.93, and 99.86 percent, respectively. Routine testing of 29,102 donations with the enhanced Abbott Plus ELISA revealed a specificity of 99.93 percent. CONCLUSION: Two Western blot‐confirmed anti‐HIV‐1‐positive samples were missed by the Abbott Recombinant ELISA but detected by the Abbott Plus and Ortho Enhanced ELISAs. The analytic sensitivity of the Ortho Enhanced ELISA was inferior to that of both Abbott ELISAs. The specificities of the Abbott Recombinant, Abbott Plus, and Ortho Enhanced ELISAs were comparable.
A randomized, controlled trial to study the efficacy and safety of C1 inhibitor concentrate in treating hereditary angioedema
Authors: Kunschak M. , Engl W. , Maritsch F. , Rosen F. S. , Eder G. , Zerlauth G. , Schwarz H. P.
Published: March 2003

BACKGROUND: No effective treatment exists in the United States for acute attacks of hereditary angioedema (HAE).

STUDY DESIGN AND METHODS: To evaluate the efficacy and safety of C1 inhibitor concentrate in treating HAE, a large primary care and referral center hospital conducted a randomized, placebo‐controlled, double‐blind trial with intent‐to‐treat analysis. Of the 36 patients enrolled in the study, 23 received treatment, and 22 completed the trial. C1 inhibitor concentrate or albumin (placebo) infusions were administered in a blind fashion to HAE patients who came to the hospital for treatment no later than 5 hours after an attack began.

RESULTS: Relief was almost twice as fast in persons receiving C1 inhibitor concentrate than in the controls: 7.62 hours (mean; SD 7.08) versus 15.35 hours (mean; SD 8.31), respectively. The difference for time‐to‐relief was highly significant (p = 0.007, Mann‐Whitney U test). The median time‐to‐relief was 6.17 hours (interquartile range 0.33‐15.35) in the treatment group and 15.35 hours (interquartile range 14.00–22.83) in the control group. Resolution of symptoms was one‐third faster in the C1 inhibitor concentrate group than in the placebo group: 23.98 hours (mean; SD 14.81) and 34.58 hours (mean; SD 13.56), respectively (p = 0.09, Mann‐ Whitney U test). Recovery of functional C1 inhibitor was 119.65 percent (mean; SD 50.80), and half‐life was 37.87 hours (mean; SD 19.75). Recovery of antigenic C1 inhibitor was 147.75 percent (mean; SD 97.68), and half‐life was 24.01 hours (mean; SD 9.70). There were no viral infections or serious adverse effects from the drug after 70 attacks in the treatment group and 96 attacks in the control group.

CONCLUSIONS: C1 inhibitor concentrate is a safe, effective treatment for acute attacks of HAE.
White cells in fresh‐frozen plasma: evaluation of a new white cell‐reduction filter
Authors: Willis J.I. , Lown J.A.G. , Simpson M.C. , Erber W.N.
Published: March 2003

BACKGROUND:Fresh‐frozen plasma (FFP) has generally been regarded as an acellular component. Recently, viable lymphocytes have been detected in this component and the question of irradiation of FFP for certain patients has been raised. Whether the numbers of white cells (WBCs) in FFP are sufficient to require WBC‐reduction of acellular components for patients receiving WBC‐reduced cellular components has not been determined. WBC numbers in FFP were examined, and the performance of a new commercial WBC‐reduction filter for FFP was assessed.

STUDY DESIGN AND METHODS:WBC numbers in plasma processed for use as FFP and in thawed FFP were counted before and after WBC‐reduction filtration by the use of flow cytometry Fast and slow filtration was used to simulate laboratory and bedside filtration, respectively. Three different methods for plasma harvesting (soft‐spin, hard‐spin, and second‐spin methods) were assessed. The filter capacity was also examined.

RESULTS:The numbers of WBCs in plasma covered a three‐log10 range (soft‐spin method, 0.04‐3.6 × 10(6); hard‐spin method, 0.47‐45.4 × 10(6); second‐ spin method, 0.4–37.2 × 10(6)). For the hard‐spin and second‐spin methods which produced the greatest plasma yields, 92 percent and 85.7 percent of bags, respectively, had counts>1 × 10(6) and 43 percent (hard‐spin method) and 45.7 percent (second‐spin method) had counts>5 × 10(6). There was no significant difference between the counts obtained in plasma and thawed FFP. The filter reduced WBC numbers to <1 × 10(5) in all but 3 of 49 bags. In the remaining three, there were <2 × 10(5) WBCs. Five bags of plasma could be processed effectively through each filter. CONCLUSION: FFP may contain WBC numbers above the threshold at which the use of WBC‐reduction filters for cellular components in some patients is necessary. Confirmation of these findings and similar investigations on plasma prepared by other methods may help in defining a role for the use of WBC‐reduction filters for FFP
Demonstration of drug‐dependent antibodies in two patients with neutrophenia and successful treatment with granulocyte–colony‐stimulating factor
Authors: Meyer O. , Gaedicke G. , Salama A.
Published: March 2003

BACKGROUND: Many drugs have been reported as being capable of inducing immune neutropenia, but the causative drug‐dependent antibodies were rarely demonstrated.

STUDY DESIGN AND METHODS: This report describes the results of serologic testing and treatment in two children with immune neutropenia related to cefotaxime and metamizole, respectively. Serum samples were tested in the presence and the absence of the drugs using the granulocyte agglutination test (GAT), the granulocyte immunofluorescence test (GIFT), and the monoclonal antibody‐specific immobilization of granulocyte antigens (MAIGA) assay.

RESULTS: The serum of one child contained cefotaxime‐dependent antibodies that were detectable by the GAT and the MAIGA assay, but not by the GIFT. The serum of the other child gave positive reactions in the GAT and GIFT due to HLA antibodies and in the MAIGA assay only in the presence of metamizole. While cefotaxime‐dependent antibody was directed against CD16, the metamizole antibody was directed against CD11b and CD35. The administration of granulocyte–colony‐stimulating factor led to an abrupt increase in circulating neutrophils in both cases.

CONCLUSION: The use of more than one technique is necessary for detection of drug‐dependent antibodies against neutrophils, and early administration of granulocyte–colony‐stimulating factor may result in fewer complications in these patients.
Effects of prestorage white cell reduction on platelet aggregate formation and the activation state of platelets and plasma enzyme systems
Authors: Devine D.V. , Bradley A.J. , Maurer E. , Levin E. , Chahal S. , Serrano K. , Gyongyossy‐Issa M.I.C.
Published: March 2003

BACKGROUND: The introduction of prestorage white cell (WBC) reduction in random‐donor platelet concentrates in Canada has increased the occurrence of particulate material in PCs. The effects of filtration on platelet activation state and the activation of plasma enzyme systems were assessed.

STUDY DESIGN AND METHODS: Particulate material was examined by light microscopy, electron microscopy, protein electrophoresis, and biochemical analysis. Thirty PCs (10 unfiltered, 20 filtered) were examined during processing and 5‐day storage for pH, platelet count and mean volume, morphology, activation marker expression, and hypotonic shock response. Complement activation, thrombin generation, and fibrinolysis were assessed by using specific enzyme immunoassays or chromogenic assays.

RESULTS: By all analyses, the particulate material appeared to be platelet aggregates. Platelets exposed to WBC‐reduction filters expressed a significantly higher level of activation markers CD62 and CD63, altered morphology, and increased platelet microparticles throughout the storage period than did unfiltered platelets. Complement activation at the C3 level was significantly increased in filtered units with little evidence of coagulation or fibrinolytic system activation.

CONCLUSION: Exposure of platelets to filters during prestorage WBC reduction increased platelet activation and mildly increased complement activation over the levels during the storage period. These alterations can contribute to the formation of irreversible platelet aggregates during processing.
Application of RHD and RHCE genotyping for correct blood group determination in chronically transfused patients
Authors: Legler T.J. , Eber S.W. , Lakomek M. , Lynen R. , Maas J.H. , Pekrun A. , Repas‐Humpe M. , Schröter W. , Köhler M.
Published: March 2003

BACKGROUND: In chronically transfused patients, conventional blood group typing may be impossible because of mixed‐field agglutination.

STUDY DESIGN AND METHODS: In 27 patients with congenital anemia and lifelong transfusion history, genotyping for D, RHD, and RHCE was performed with polymerase chain reactions. These results were compared with the blood group typing results documented in the medical record.

RESULTS: Two of 27 cases had been typed D‐negative by serologic tests and D‐positive by genotyping. In 20 patients, the CDE formula had been determined serologically according to the medical record; 4 of these patients were Cc by serologic tests and C/C by genotyping. One patient typed ee by serologic tests, and genotyping revealed heterozygosity (E/e).

CONCLUSION: In patients with a lifelong transfusion history, serologic blood group determination may be impossible, and pretransfusion test results are not always available or reliable. In whites, Rh‐matched transfusions are possible with genotyping. The genetic background of the RH genes has to be elucidated in other ethnic groups, such as in black patients with sickle cell disease, before genotyping can be applied without restriction.
Plateletpheresis efficiency: a comparison of the Spectra LRS and AMICUS separators
Authors: Benjamin R. J. , Rojas P. , Christmas S. , Neal J. , Broughton S. , Burgio C. , Barrett B. , Churchill W.H.
Published: March 2003

BACKGROUND: Optimizing the yields of platelet‐pheresis by increasing collection efficiencies may provide several benefits: to patients, by increasing the dose in single‐donor platelet (SDP) units; to the collection center, by increasing the percentage of components that may be split into double units; and/or to the donor, by reducing the duration of donation.

STUDY DESIGN AND METHODS: A prospective, randomized study was undertaken to compare the efficiency of platelet collection in paired donations by 21 donors on two cell separators (Spectra LRS, version [v] 5.1; and AMICUS, v2.37). The order of donation was randomly assigned; donations were performed at least 2 weeks apart. A fixed blood volume (4000 mL) was processed by utilizing standard protocols. Findings were confirmed in a retrospective, matched study that compared the two separators by using fixed collection times (90 min).

RESULTS: The AMICUS and Spectra LRS separators consistently produced white cell‐reduced (<1× 106 white cells) components. The AMICUS harvested 32 percent more platelets on average (median, 4.9 × 1011; range, 1.5–8.7 × 1011) than the Spectra LRS (median, 3.7 × 1011; range, 2.1‐7.7 × 1011) (p = 0.03), with mean times of 71.5 and 66.0 minutes (p = 0.03), respectively, to process 4000 mL. Mild donor reactions tended to be more common on the AMICUS separator, which used significantly more ACD (median 482 mL vs. 389 mL; p<0.0001) than on the Spectra LRS.

CONCLUSIONS: The AMICUS separator harvested more platelets per unit of blood volume processed than the Spectra LRS. Possible benefits include increased dose in each single‐donor unit, increased double‐unit harvests, and/or shorter donation time.
Sensitivity and specificity of human T‐lymphotropic virus (HTLV) types I and II polymerase chain reaction and several serologic assays in screening a population with a high prevalence of HTLV‐II
Authors: Liu H. , Shah M. , Stramer S.L. , Chen W. , Weiblen B.J. , Murphy E.L.
Published: March 2003

BACKGROUND: Since 1988, all blood donations in the United States have been screened for antibodies to human T‐lymphotropic virus type I (HTLV‐I). However, the sensitivity of current serologic tests for the detection of HTLV type II (HTLV‐II) antibodies and the diagnostic utility of direct tests for HTLV‐I and ‐II using polymerase chain reaction (PCR) are poorly defined.

STUDY DESIGN AND METHODS: Five hundred sixty‐nine HTLV‐I‐ or ‐II‐seropositive and 687 age‐ and sex‐matched seronegative samples from a high‐risk population at an inner‐city emergency department were selected. All samples were tested with four HTLV enzyme immunoassays (EIAs), one Western blot assay and one type‐specific Western blot assay, one HTLV type‐specific EIA, and a research HTLV‐I/II PCR kit.

RESULTS: Sensitivity of the various EIAs ranged from 95.1 to 99.5 percent, and specificity ranged from 97.2 to 99.4 percent. PCR performed in duplicate without selective retesting had lower sensitivity (85.1%) and specificity (88.0%). However, PCR detected 20 (3.2%) HTLV‐I‐positive and 47 (7.5%) HTLV‐II‐positive samples among the 627 samples that were negative in all EIAs. The type‐specific EIA and PCR assay had the highest rate of concordance in classifying samples as either HTLV‐I or ‐II, with the type‐specific EIA and type‐specific Western blot having the next highest rates of concordance.

CONCLUSION: In this sample set from a population at high risk for HTLV‐II, screening with HTLV‐I/II PCR had lower sensitivity and specificity than that with EIAs. However, 4.1 to 10.8 percent of samples were PCR positive but seronegative for HTLV‐I or –II, and their true infection status remains undetermined.