bluuebunny/crossref_metadata_2025 huggingface dataset:
Prophylactic granulocyte transfusions: results of a randomized controlled trial in patients with acute myelogenous leukemia
Authors: Ford J. M. , Cullen M. H. , Roberts M. M. , Brown L. M. , Oliver R. T. D. , Lister T. A.
Published: December 2004

A prospective randomized and controlled clinical trial of prophylactic granulocyte transfusions was conducted in an attempt to reduce the mortality from infection in newly diagnosed adults with acute myelogenous leukemia. Granulocytes were harvested from normal donors by cytapheresis, and transfused patients received a median of 1.45 × 10(10) granulocytes (range 0.28–3.45 × 10(10)) on alternate days during marrow aplasia caused by initial induction chemotherapy. Transfusion were started if the absolute peripheral granulocyte count was less than 500 microliters. Thirteen patients received from one to 12 granulocyte transfusions, and 11 control patients received no granulocytes. No significant advantage was demonstrated in the transfused patients compared with controls with regard to the following: 1) deaths due to infection, 2) reduction in the frequency of febrile episodes, 3) delay in the onset of fever, 4) reduction in the length of febrile episodes, or 5) reduction in the frequency of proven infection.
Selection of compatible kidney donors for recipients with multiple cytotoxic antibodies
Authors: Larsen A. E.
Published: December 2004

Cytoxic antibodies developed in 33 (14.6%) of 226 renal transplant candidates while awaiting transplantation. Antibody formation was not associated with primary kidney disease, nor was the frequency of antibody formation increased following deliberate transfusion. Analysis of the HLA types of panel cells against which cytotoxic sera failed to react predicted donor kidney antigens which would be acceptable to the recipient. Crossmatching with all stored, reactive serum samples as well as a current sample, is imperative.
Granulocyte transfusions and outcome of alloimmunized patients with gram‐negative sepsis
Authors: Dahlke M.B. , Keashen M. , Alavi J.B. , Koch P.A. , Eisenstaedt R.
Published: December 2004

The effect of white cell alloimmunization on patient outcome during gram‐negative sepsis treated with granulocyte transfusions was studied. Twenty‐five episodes of sepsis were observed; 19 were associated with resolution of sepsis and six with continuing sepsis and death. Compatibility testing included the granulocyte indirect immunofluorescence test and the lymphocytotoxicity assay. The number of compatible and incompatible granulocyte transfusions determined by the indirect immunofluorescence test compared with patient outcome was significant (X2 = 44, p less than 0.001). The same comparison with the lymphocytotoxicity assay was not significant (X2 = 3, p greater than 0.05). The duration of the granulocytopenia after the first positive blood culture was 10 days or longer in 13 patients, and 12 of the 13 survived. The duration of the granulocytopenia was less than 10 days in 12 patients, and five of the six deaths occurred in this group. These five patients died without evidence of bone marrow recovery and with persistent gram‐negative sepsis. Each had granulocyte‐specific antibody of broad specificity and none received five consecutive compatible transfusions.
The biochemical effects of CPDA‐2‐drawn red blood cells of delayed refrigeration prior to component preparation
Authors: Moore G.L. , Ledford M.E. , Merydith A.
Published: December 2004

The effects of an 8‐hour hold at 22 degrees C prior to component preparation were evaluated in a split‐bag study using nine units of blood preserved in citrate‐phosphate‐dextrose‐adenine (CPDA‐2). Each unit was divided in half, platelet‐rich plasma removed at 0 or 8 hours, respectively, and the half units of red blood cells stored at 4 degrees C for 42 days. The only red blood cell metabolic differences seen in the bags held 8 hours (compared to those not held) were a 21 percent rise in adenosine triphosphate, which was not significant after 14 days of storage, and a 33 percent loss of 2,3‐diphosphoglycerate which resulted in a loss curve similar to that seen with acid‐citrate‐ dextrose blood. The logistic advantages seem to warrant an 8‐hour holding period for red blood cells drawn in CPDA‐2.
The effect of centrifugation time on cellular residues in fresh plasma
Authors: Lau P. , Sererat M. , Bartkowiak M. V. , Moore V.
Published: December 2004

Cellular residues were present in all fresh plasma prepared by a standard technique with centrifugation times of 5, 7, or 10 minutes. The amounts of cellular residues in the plasma were inversely proportional to the time of centrifugation. Platelet contamination was statistically different in the three times tested. In contrast, the residual amounts of erythrocytes and leukocytes were significantly different only when 5‐ and 10‐minute centrifugation times were compared. TRANSFUSION 1983;22:000‐000.
Modification of acid stromal elution method for complete recovery of bound antibodies
Authors: ARASZKIEWICZ P. , HUFF S.R. , SZYMANSKI I.O.
Published: December 2004

ABSTRACT: Erythrocyte‐bound antibodies could be eluted from stroma almost completely using a modification of the method of Jenkins and Moore.(1) In the modified method, eight volumes of 0.1M glycine, pH 3.0, were added to one volume of stroma and incubated at 45°C for 30 minutes. This method was optimal for autoantibodies and all allo‐antibodies studied.
Dimethyl sulfoxide: effects on function of fresh platelets and on the viability of platelets in storage
Authors: Fratantoni J. C. , Poindexter B. J.
Published: December 2004

Dimethyl sulfoxide (DMSO) is used as a cryoprotective agent when platelets are frozen. We examined the effect of DMSO (0.1 to 10%) on platelet aggregation, release, and prostaglandin synthesis (as indicated by malondialdehyde formation) in response to thrombin, collagen, arachidonic acid and calcium ionophore. Inhibition was observed at the lowest levels of DMSO, varied with the type of stimulus, and was reversed by washing the platelets. Inhibition of aggregation, release, and malondialdehyde formation were dose‐dependent with thrombin or collagen. DMSO did not inhibit malondialdehyde formation stimulated by arachidonic acid, nor did it consistently inhibit any function stimulated by calcium ionophore. When platelets were stored as platelet‐rich plasma at 20 to 24°C for 48 hours, with and without 5 percent DMSO, and subsequently washed, the platelets stored with DMSO were more reactive in vitro. These results indicate that platelet function inhibition by DMSO not only is reversible, but protects the platelets during storage. The factor limiting the use of DMSO in platelet storage is potential systemic toxicity, not its effects on platelets.
Phosphate ion exchange resin used in the liquid preservation of baboon red cells
Authors: Valeri C. R. , Ouellet R. , Mostacci J. , Harmening‐Pittiglio D. , Vecchione J. J.
Published: December 2004

Baboon whole blood, collected in 14 percent citrate‐phosphate‐dextrose anticoagulant solution In plastic bags was stored in 100‐ml aliquots at47deg; C for 28 days in the presence or absence of 0.75 grams of phosphate anlon exchange resin. In vitro and in vivo measurements after autologous transfusions were made to determine whether the phosphate anion exchange resin had any beneficial effect on the blood during storage. The in vitro measurements of red cell 2,3‐diphosphoglycerate and P50 were higher throughout the 28 days of storage at 4° C in the blood stored in the phosphate anion exchange resin. After autologous transfusions in six baboons of red cells prepared from whole blood stored at 4° C for 21 days, the 24‐hour posttransfusion survival values were 86 ± 6 percent (mean ± SD) in the presence of resin and 83 ± 6 percent in the absence of resin. In five other baboons, red cells prepared from 28‐day‐old blood showed a mean 24‐hour posttransfusion survival of 82 ± 4 percent in the presence of resin and 75 ± 4 percent in the absence of resin. The addition of a phosphate anion exchange resin to the citrate‐phosphate‐dextrose anticoagulant provided better maintenance of red cell 2,3‐diphosphoglycerate concentrations and P50 levels during storage of whole blood at 4° C, and red cells prepared from whole blood stored in this solution had better oxygen transport function than red cells prepared from blood stored without resin. Red cell adenosine triphosphate concentrations and 24‐hour posttransfusion survival values were similar whether or not the anticoagulant contained resin.
Incompatibility associated with the bite of a brown recluse spider (Loxosceles reclusa)
Authors: Hardman J. T. , Beck M. L. , Hardman P. K. , Stout L. C.
Published: December 2004

Red cell samples from a patient who was suffering from massive hemolysis due to a brown recluse spider (Loxosceles reclusa) bite were found to be reactive by indirect antiglobulin test with most ABO‐compatible serum samples. Spider venom, enzymes related to those in spider venom, and antisera to plasma proteins and Rh antigens were used to investigate the unusual reactivity of the patient's cells. IgG was detected on the patient's cells by indirect antiglobulin tests. Cells treated with brown recluse spider venom or phosphatidylcholine phosphatidohydrolase reacted similarly. These findings suggest that sphingomyelinase D, which has been identified in brown recluse spider venom, may be related to the unusual reactivity of the patient's cells. Unexpected reactions were observed when venom‐treated cells were tested with Rh antibodies: O negative cells absorbed and eluted anti‐D from Rh immune globulin; E negative cells were reactive with a commercial anti‐E reagent.
In vivo survival of K:18 red cells in a recipient with anti‐K 18
Authors: Barrasso C. , Baldwin M. L. , Drew H. , Ness P. M.
Published: December 2004

Red cell survival, IgG subclass, and mononuclear phagocyte assay studies were performed on a patient with an antl‐K + K18 described previously. The 51Cr survival study with Kell negative K:18 red cells showed 76.6 percent survival at 75 minutes with 30.7 percent survival at 24 hours. The anti‐K18 was characterized as lgG4 + lgG1. The mononuclear phagocyte assay was 16 percent when the serum was tested with K:18 cells. Among 54,450 ABO compatible donor units tested with this serum, no K:18‐ unit was found. These studies were undertaken to evaluate the clinical significance of the antibody when red cell support for a chemotherapy course was considered. Our data suggest that transfusion with K:18+ blood would be ineffective and could be used only to provide red cell support in an emergency should compatible units be unavailable.
A summary of the results of screening for HLA antibodies for one year
Authors: LARSEN A. E.
Published: December 2004

ABSTRACT: In one year, 748 serun specimens from obstetric patients were screened for HLA antibodies. Of the 284 showing antibody activity, 179 were nonspecific and 68 were multispecific. Thirty‐seven (5%) were found to be monospecific, representing 11 specificities, of which the most common were anti‐B7, ‐B8, ‐A2, and ‐B12. Recovery of antisera for reagent use was financially feasible.
Post‐perfusion syndrome due to Epstein‐Barr virus
Authors: McMonigal K. , Horwitz C. A. , Henle W. , Henle G. , Lawton J. , Polesky H. , Peterson L.
Published: December 2004

The post‐perfusion syndrome is rarely due to Epstein‐Barr virus (EBV), because most adult patients already carry protective neutralizing antibodies in their sera. Data are presented from two patients in whom heterophil‐antibody‐positive infectious mononucleosis developed 35 and 48 days after open‐heart surgery. The diagnoses of their predominantly febrile illnesses were delayed because of late appearance of atypical lymphocytes and heterophil antibodies. EBV‐specific antibody responses were also delayed, with peak titers appearing several weeks or months after onset. The differences in evolution of the present cases, compared to those of classic infectious mononucleosis, were presumably due to different portals of entry (blood stream versus oropharynx).
The effect of blood cooling prior to granulocyte collection
Authors: Bensinger W. I. , Williams B. M. , Friedman L. I. , Buckner C. D. , Clift R. A.
Published: December 2004

A study was designed to test the hypothesis that cooling blood before it enters a continuous‐flow blood cell separator would result in increased granulocyte yields. Fifty‐four granulocyte collections performed with cooling were compared with 58 collections performed on alternate days in the same donors. In blood cooled to 15°C we were unable to demonstrate improved yields of granulocytes.
Hepatitis B surface antigen in blood donors
Authors: Jayaprakash P. A. , Shanmugam J. , Hariprasad D.
Published: December 2004

Of 8085 volunteer donors attending the blood bank at SCTIMST screened for hepatitis B surface antigen (HBsAg) carrier state by counterimmunoelectrophoresis, 103 (1.27%) were HBsAg positive. The personal data of donors showed a higher rate of HBsAg among men than women and in the age group of 21 to 30 years than in the other age groups. A significantly higher rate was noted among donors belonging to the lower socioeconomic group (p < 0.05).
Alterations of lymphocyte responsiveness in Guillain‐Barré syndrome
Authors: Glassman A. B. , Bennett C. E.
Published: December 2004

Various antibodies and immune complexes have been demonstrated in patients with Guillain‐Barre syndrome (GBS). These factors may be part of the pathogenesis of the disease process through immune complex deposition and/or suppression of the immune system. Four patients with typical GBS were treated by plasma exchange. Aliquots from the first 100 ml of plasma removed from each patient were incubated with cultures of normal human peripheral lymphocytes that subsequently were exposed to mitogens, followed by incubation with tritiated thymidine. A decrease (p < 0.001) in response to each of the mitogens was noted after incubation with patient plasma. Peripheral lymphocytes were taken from the patients before and after plasma exchange. An increase (p <0.01) in DNA synthesis following mitogen exposure was observed after plasma exchange in each patient. All patients exhibited subjective and objective clinical improvement. Plasma exchange for GBS appeared to remove a lymphocyte suppressor factor, which was characterized by column fractionation. The suppressor factor was determined to be a substance of approximately 10,000 to 12,000 daltons that reproducibly decreased the responsiveness of normal lymphocytes to mitogens.
Effect of cardiopulmonary bypass on plasma fibronectin, IgG, and C3
Authors: Gandhi J. G. , Salm T. Vander , Szymanski I. O.
Published: December 2004

The effect of cardiopulmonary bypass operations on the concentration of plasma fibronectin, IgG, and C3 was studied in 28 patients. During cardiopulmonary bypass operations, plasma IgG, and C3 decreased to about 45 percent of their preoperative concentrations, probably as a result of hemodilution and blood loss. On the average, plasma fibronectin decreased significantly more (p < 0.002), to 32 percent of its preoperative concentration, probably as a result of hemodilution, blood loss, and consumption. We assumed that consumption of fibronectin occurred when its concentration decreased more than the concentrations of IgG and C3. Interestingly, consumption of fibronectin during bypass operations was not demonstrated in seven of the 28 patients who did not require blood products within the 24‐hour period after operation. We suggest that consumption, rather than dilution, of plasma fibronectin might be clinically important. A significant (p < 0.002) negative correlation was observed between the length of the bypass and the concentration of plasma fibronectin during this 24‐hour period.

We also studied the rate of regeneration of fibronectin, IgG, and C3 in 12 of these patients. The concentrations of fibronectin and C3 were normal by the fifth postoperative day. The rate of regeneration of IgG, however, was slower than that of fibronectin and C3.
Normal bone marrow as a source of granulocytes for transfusion
Authors: Aghai E.
Published: December 2004

A method for obtaining granulocytes for transfusion by direct aspiration of bone marrow from normal donors is described. The bone marrow is the major storage reserve for granulocytes with as many as 60 times more granulocytes than in the peripheral blood. With current methods of leukapheresis from peripheral blood, steroids, hydroxyethyl starch, and anticoagulants are given to the donor and his processed blood is returned. Bone marrow aspiration spares the donor such treatment. In addition, no costly machine is necessary and the whole procedure takes less than 2 hours. Due to minimum manipulation, cell viability is not impaired. Furthermore, some committed granulocyte stem cells, as well as nonmature granulocytes, are transfused. These cells have the potential to mature to efficient phagocytes in the patient's circulation. TRANSFUSION 1983;23:496–499.
Quantification of the peripheral blood colony forming unit‐culture rise following chemotherapy Could leukocytaphereses replace bone marrow for autologous transplantation?
Authors: Stiff P.J. , Murgo A.J. , Wittes R.E. , DeRisi M.F. , Clarkson B.D.
Published: December 2004

Because hematopoietic stem cells circulate in the peripheral blood (PB), the possibility exists for using leukocytaphereses in place of marrow for autologous transplantation. Normal stem cells, measured by the colony forming unit‐culture (CFU‐C) assay, usually in extremely small numbers in PB, are increased afteraggressive chemotherapy. Patients with small‐cell undifferentiated carcinoma of the lung had PB assayed for CFU‐C numbers during induction chemotherapy. The median blood CFU‐C concentration increased as much as 13.3‐fold compared to pretreatment values following the third cycle of chemotherapy. The CFU‐C peaks occurred at the same time as a relative monocytosis in the PB, with monocytosis percentages often 30 percent or higher. Compared to an average CFU‐C dose obtained from a typical marrow procedure done under general anesthesia (4.0 × 104 CFU‐C/kg), the anticipated CFU‐C yield obtained from a single cytapheresis performed at the CFU‐C peak after the third course of chemotherapy (2.9 × 104 CFU‐C/kg) suggests that as few as two‐cytaphereses could provide sufficient stem cells to reconstitute hematopoiesis following lethal myelosuppression.
Trial of a combined test for the detection of HBsAg and anti‐HBc
Authors: Wylie J.S. , Mankikar S.D. , Buchner B.K. , Moore B.P.L.
Published: December 2004

A combined hepatitis test (Ausria/Core, Abbott Laboratories, Wiesbaden, West Germany) detects hepatitis B surface antigen by radioimmunoassay and antibody to the core antigen by enzyme immunoassay using a single bead. It was found to have comparable sensitivity to commercially available tests from the same manufacturer for each component. Reproduci‐bility was satisfactory for both components. The combined test is simpler and cheaper to perform than two separate tests.
Activation of the complement system by different autologous transfusion devices: an in vitro study
Authors: Tylman Maria , Bengtson Jan Peter , Bengtsson Anders
Published: December 2004

BACKGROUND : The aim of the present investigation was to study whether autologous transfusion devices activate the complement system and whether complement‐activated blood is more vulnerable to further activation during processing.

STUDY DESIGN AND METHODS : Forty‐eight blood units were randomized to be processed by one of three different salvage systems: Group 1 underwent whole blood filtration (hemofiltration) (n = 16); Group 2 underwent continuous processing, saline washing, and centrifugation (CATS, Fresenius AG ) (n = 16); and Group 3 underwent saline washing and centrifugation (Cell‐Saver, Haemonetics Corp.) (n = 16). Eight blood units for each system were activated with cobra venom factor (CVF) at a concentration of 0.2 U per mL whole blood before processing. C activation was studied by determinations of C4d, Bb, C3a, and SC5b‐9. Samples were drawn from whole blood, processed blood, and the waste bags.

RESULTS : The concentrations of Bb, C3a, and SC5b‐9 in whole blood after activation with CVF were significantly elevated compared to blood that was not activated (p < 0.01). Processed blood from hemofiltration contained significantly higher levels of complement‐split products than techniques that use washing and centrifugation. The concentrations of SC5b‐9 in blood processed by hemofiltration were higher in the experiments with CVF activation (p < 0.05).

CONCLUSION : The tested autologous transfusion systems did not themselves activate the complement system, and complement‐activated blood was not more vulnerable to further activation during processing. A blood‐salvaging technique that used washing and centrifugation reduced elevated concentrations of complement‐split products, whereas hemofiltration did not.
CMV DNA is rarely detected in healthy blood donors using validated PCR assays
Authors: Roback John D. , Drew W. Lawrence , Laycock Megan E. , Todd Deborah , Hillyer Christopher D. , Busch Michael P.
Published: December 2004

BACKGROUND : Although serologic screening or WBC reduction of blood components can reduce the incidence of transfusion‐transmitted CMV (TT‐CMV) infection, ‘breakthrough’ cases of TT‐CMV still occur and may produce serious sequelae. NAT of blood components for CMV DNA has been proposed to further reduce the risks of TT‐CMV. However, large‐scale studies to determine the utility of validated CMV NAT assays for donor screening have not been reported.

STUDY DESIGN AND METHODS : Coded whole‐blood samples (n = 1000) were tested for the presence of CMV DNA using two CMV PCR assays previously validated in a multicenter trial (a nested PCR assay directed at the CMV UL93 open‐reading frame and the Roche Monitor assay). Corresponding plasma samples were tested in parallel for the presence of anti‐CMV using other assays (Abbott CMV EIA and Fujirebio/Olympus CMV particle agglutination assays).

RESULTS : In total 416 and 514 of the samples tested as CMV‐seropositive and ‐seronegative, respectively, by both antibody assays. The remaining 70 samples had discrepant serology results. Only 2 of the 1000 samples (both seropositive) had reproducibly detectable CMV DNA (positive in at least three of four replicates). CMV DNA was not reproducibly detected in seronegative samples or in samples with discrepant serology results.

CONCLUSIONS : Although previous investigations showed frequent detection of CMV DNA in healthy CMV‐seropositive (and some seronegative) blood donors, these studies were relatively small and the performance characteristics of their assays were difficult to evaluate. In contrast, the present large cross‐sectional study of US donors utilized two previously validated PCR assays and demonstrated that CMV DNA is only rarely detectable in seropositive donors. Thus, the use of CMV PCR assays with optimal performance characteristics did not increase the detection of potentially infectious blood components beyond that provided by current serologic screening assays.
Systematic analysis of the ABO gene diversity within exons 6 and 7 by PCR screening reveals new ABO alleles
Authors: Seltsam Axel , Hallensleben Michael , Kollmann Anke , Burkhart Jürgen , Blasczyk Rainer
Published: December 2004

BACKGROUND: Mutations critical for ABO blood group phenotypes have predominantly been found in exons 6 and 7 of the ABO gene, both of which encode the catalytic domain of ABO glycosyltransferase. To design rapid and reliable ABO genotyping assays, a profound knowledge of the prevalent alleles is required and a reliable sequence database needs to be established.

STUDY DESIGN AND METHODS: A PCR screening system was established consisting of 102 different PCRs, each specific for a single nucleotide (nt) variation. The primer mixes were developed to walk from the 5′ to the 3′ end of exons 6 and 7 of the ABO gene to screen for nt mutations at 50 known polymorphic sites. A total of 109 unrelated individuals with common and rare ABO characteristics were screened. All blood samples in which the PCR results were inconclusive or inconsistent with the ABO phenotypes were subjected to sequence analysis of exons 6 and 7.

RESULTS: The results of PCR screening were conclusive and consistent with the ABO phenotypes in 90 cases. In the remaining 19 cases, PCR screening revealed unusual allele combinations or amplification results that were incompatible with known ABO allele combinations or subgroups predicted by serologic analysis. In these 19 cases, sequencing revealed new ABO alleles (one ABO*Ael allele, one ABO*B(A) allele and two ABO*O alleles) in two individuals with common and seven individuals with variant ABO phenotypes.

CONCLUSION: This PCR screening strategy is an effective tool for obtaining deeper insight into the ABO gene diversity and diversification and may be useful to increase the quality of the ABO sequence database.
RBC transfusion requirements after allogeneic marrow transplantation: impact of the before‐transplant Hb level on transfusion and early survival
Authors: Xenocostas Anargyros , Yee Angelina , Wong Cindy J. , Sutton David M.C. , Lipton Jeffrey H. , Kiss Thomas L. , Messner Hans A.
Published: December 2004

BACKGROUND: Most patients undergoing allogeneic marrow transplantation (alloBMT) require transfusions of RBCs. A retrospective analysis was performed to evaluate the utilization and risk factors for RBC transfusions including age and sex of recipient, HLA matching between donor and recipient, disease status at time of BMT, the occurrence of GVHD, ABO blood group compatibility, the source of progenitor cells and the Hb level before BMT (PT‐Hb).

STUDY DESIGN AND METHODS: Data from 519 consecutive patients receiving transplants between January 1995 and March 2000 were reviewed. The number of RBC transfusions was determined for the following periods: 0 to 60, 61 to 120, and 121 to 180 days after BMT.

RESULTS: The transfusion requirements were greatest during the first 60 days after BMT and decreased with time. The total number of units transfused to this cohort of patients was 5398, of which 3505 units were utilized within the first 2 months. The mean number ± SD of units transfused per patient from 0 to 60 days was 6.8 ± 6.4; 61 to 120 days, 3.2 ± 5.5; and 121 to 180 days, 2.0 ± 4.6. An increased transfusion requirement was associated with lower PT‐Hb, major ABO mismatch between donor and recipient, BMT in patients with more advanced disease, use of unrelated donors, older age, and female sex by Spearman's correlation analysis. The source of progenitor cells and the development of GVHD did not influence transfusion requirements. Increased mortality during the 6‐month period after transplant was associated with lower PT‐Hb, use of unrelated donors, advanced disease status at BMT, and sex by Cox regression analysis. In a multivariate model, PT‐Hb remained significant when controlling for the other risk factors.

CONCLUSION: The PT‐Hb was identified as an independent risk factor for RBC transfusions during alloBMT. As well, a lower PT‐Hb was found to be an independent risk factor for increased mortality during the 6‐month study period.
MP4, a new nonvasoactive PEG‐Hb conjugate
Authors: Vandegriff Kim D. , Malavalli Ashok , Wooldridge Jerod , Lohman Jeff , Winslow Robert M.
Published: December 2004

BACKGROUND: Vasoconstriction has been an obstacle to clinical development of Hb‐based O2 carriers. It is proposed that this limitation can be overcome by increasing molecular size and oxygen affinity.

STUDY DESIGN AND METHODS: Surface‐modified Hb (MP4) was designed, whose properties are consistent with the theory that cell‐free Hb engages autoregulatory vasoconstrictive responses to Hb diffusion in the plasma space (“facilitated diffusion”). Human Hb was modified by reaction first with 2‐iminothiolane to add sulfhydryl groups and then with monofunctional maleimide‐ activated 5‐kDa PEG.

RESULTS: MP4 was found to have a molecular weight of 90 kDa, a molecular radius increased relative to native Hb (9.3 ± 1.4 vs. 3.2 nm), high oxygen affinity (p50∼ 5‐6 mmHg), and a Bohr effect approximately half that of native human Hb (–0.24Δlogp50/ΔpH). At 4.2 g per dL in Ringer's lactate, its viscosity was 2.5 cP, and its oncotic pressure was 50 mmHg. The t50 of 14C‐MP4 in rats was approximately 24 hours. No significant elevation in mean arterial pressure was observed.

CONCLUSION: MP4 appears to be free of a pressor effect, a major limitation to the development of a safe and effective RBC substitutes in the past.
Apoptotic activity in stored human platelets
Authors: Perrotta Peter L. , Perrotta Cheryl L. , Snyder Edward L.
Published: December 2004

BACKGROUND: Platelets possess some of the machinery required for apoptotic cell death. However, disruption of mitochondria function, implicated in several models of cell death, has not been extensively studied in platelets. Mitochondrial viability and several other measures of apoptotic death in stored and experimentally stressed platelets were evaluated.

MATERIALS AND METHODS: Platelet mitochondrial transmembrane potentials (Δψm) were studied by staining platelets with JC‐1, a dye that fluoresces at different wavelengths based on the state of mitochondrial polarization. Annexin V binding, a measure of phosphatidylserine (PS) exposure, and CD62P expression, an indicator of platelet activation, were determined by flow cytometry. Caspase‐3 activity was measured with an enzyme assay and by Western blotting. Experimental platelet stressors included storage for 7 days, azide exposure, calcium ionophore stimulation, and plasma deprivation.

RESULTS: As measured by flow cytometry, Δψm values were similar in freshly drawn platelets and in platelet concentrates stored for up to 7 days. However, compared to fresh platelets, stored platelet concentrates had significantly increased PS exposure (3.1 vs. 5.1%, p = 0.015), CD62P expression (6.5 vs. 13.5%, p = 0.0067), and caspase‐3 activity. Azide exposure, which decreased ATP release 20 to 30 percent, did not affect the Δψm. Stressed platelets exhibited higher degrees of mitochondrial depolarization in response to calcium ionophore stimulation than platelets that were not stressed. Plasma deprivation also resulted in significant alterations in Δψm, PS exposure, and CD62P expression.

CONCLUSIONS: Platelet mitochondria maintain Δψm when stored for up to 7 days under standard blood bank storage conditions. Therefore, changes in platelet mitochondria Δψm do not correlate with downstream markers of apoptotic death such as caspase activation and PS exposure.
A closed and single‐use system for monocyte enrichment: potential for dendritic cell generation for clinical applications
Authors: Rouard Helene , Leon Anne , De Reys Stef , Taylor Linda , Logan Josephine , Marquet Jeanine , Jouault Helene , Loper Kathy , Maison Patrick , Delfau‐Larue Marie‐Helene , Beaujean Françoise , Farcet Jean‐Pierre , Noga Stephen J.
Published: December 2004

BACKGROUND: This study evaluated the ability of a modified cell separator (Cobe Spectra Apheresis) system to isolate monocytes (MOs) by elutriation. The evaluation was performed in two independent international laboratories. The capacity of collected MOs to differentiate into dendritic cells (DCs) was also assessed.

STUDY DESIGN AND METHODS: MNCs from platelet apheresis residues were elutriated on a modified cell separator (Cobe Spectra Apheresis system) using a custom disposable set. Cells were separated according to their size and density. Recovery and purity of the collected cell product were evaluated by impedance counting and flow cytometry. DCs were differentiated in culture from the elutriated MOs and characterized by their surface markers and stimulatory capacity in a mixed WBC reaction assay.

RESULTS: Six apheresis mononuclear cell products were used by each laboratory. The separation was achieved in less than 1 hour. Collected MOs had the potential to differentiate into DCs.

CONCLUSION: The modified cell separator is an easy and fast device to obtain highly enriched MOs with a DC differentiation potential. The system is closed and employs a single‐use disposable set and is more amenable to good tissue practice. This method could become a valuable tool for DC‐based active immunotherapy.
Comparative sensitivity of HBV NATs and HBsAg assays for detection of acute HBV infection
Authors: Biswas Robin , Tabor Edward , Hsia Chu Chieh , Wright David J. , Laycock Megan E. , Fiebig Eberhard W. , Peddada Lorraine , Smith Richard , Schreiber George B. , Epstein Jay S. , Nemo George J. , Busch Michael P.
Published: December 2004

BACKGROUND: A study was designed to estimate relative analytic sensitivity and window‐period (WP) closure and to project incremental yield of newer HBsAg tests, pooled‐sample NAT, and single‐sample NAT, compared to currently licensed HBsAg tests.

STUDY DESIGN AND METHODS: HBV DNA and HBsAg test results for 23 HBV seroconversion (SC) panels were first analyzed to construct a model of primary HBV viremia. One‐hundred representative samples were then selected from 10 panels and coded with 28 analytical controls. All 128 samples were tested by seven HBsAg tests and by four pooled‐sample and three single‐sample NAT assay formats. Results were analyzed to obtain differential times to HBV detection and combined with HBV incidence rates to project comparative yields.

RESULTS: HBV doubling time during the ramp‐up phase was estimated at 2.56 days. HBsAg concentrations at cutoff for new tests ranged from 0.07 to 0.12 ng per mL, compared with 0.13 to 0.62 ng per mL for licensed tests. Estimated viral load at cutoff ranged from 102 to 267 IU per mL for new tests and from 363 to 1069 IU per mL for licensed tests. HBsAg tests detected 31 to 63 percent of early ramp‐up phase samples in the 100‐member seroconversion panel study, while pooled‐sample NAT detected 55 to 71 percent and single‐sample NAT, 82 to 99 percent. Compared with currently licensed HBsAg assays, newer HBsAg assays would reduce the WP by 2 to 9 days; pooled‐sample NAT would reduce the WP by 9 to 11 days; and single‐sample NAT would reduce the WP by 25 to 36 days.

CONCLUSION: Newer HBsAg tests would be expected to detect an additional 15 to 21 infected units per 107 donations, compared to licensed HBsAg tests. Sensitivity, WP closure, and yield projections for newer HBsAg assays and pooled‐sample NAT are comparable. Single‐sample NAT would increase yield by 13 to 15 units per 107 donations over pooled‐sample NAT and newer HBsAg assays and by 35 to 50 units per 107 donations over currently licensed HBsAg assays.
Wash‐out of DMSO does not improve the speed of engraftment of cord blood transplantation: follow‐up of 46 adult patients with units shipped from a single cord blood bank
Authors: Nagamura‐Inoue Tokiko , Shioya Mika , Sugo Michiko , Cui Yan , Takahashi Atsuko , Tomita Satomi , Zheng Yizhou , Takada Kei , Kodo Hideki , Asano Shigetaka , Takahashi Tsuneo A.
Published: December 2004

BACKGROUND:  Prolonged periods of marrow hypoplasia have been a problem in cord blood transplantation. DMSO is thought to produce osmotic shock to the progenitors when the thawed cells are infused into the patients. To solve this problem, a 2× dilution method originally developed in the New York Blood Center showed earlier myeloid engraftment,1 although follow‐up clinical studies have not performed.

STUDY DESIGN AND METHODS:  To clarify the influence of the removal of DMSO by this method on the speed of engraftment in unrelated cord blood transplantation, 46 adult patients with cord blood units processed by the Tokyo Cord Blood Bank from September 1998 to March 31, 2002 were studied. Twenty‐four patients received 2.6 ± 0.71 × 107 nucleated cells per kg without washing (nonwashed group), while 22 patients were received 2.7 ± 0.52 × 107 nucleated cells per kg after 2× dilution washing (washed group).

RESULTS:  The cumulative incidence of engraftment was not significantly different between the two groups. Median neutrophil recovery (≥5 × 109/L) in the nonwashed and washed groups was 26 and 25 days, respectively, and the median platelet recovery (≥20 × 109/L) in patients with myeloid engraftment was 44 and 40 days, respectively (NS). On the other hand, the doses of CFCs and CD34+ cells showed the influence on myeloid and platelet recovery.

CONCLUSION:  A 2× dilution after thawing cord blood did not result in the improvement of myeloid engraftment speed.
Detection of bacteria in WBC‐reduced PLT concentrates using percent oxygen as a marker for bacteria growth
Authors: Ortolano Girolamo A. , Freundlich Lawrence F. , Holme Stein , Russell Rosalind L. , Cortus Mary Anne , Wilkins Karen , Nomura Hiromi , Chong Chiyong , Carmen Raleigh , Capetandes Anthony , Wenz Barry
Published: December 2004

BACKGROUND:  The risk of receiving a PLT concentrate (PC) contaminated with bacteria may be 1000‐fold greater than that of pathogenic viral transmission, yet surveillance for this risk is not generally practiced. A novel bacteria detection system (BDS) that overcomes the limitations of current systems is described. The BDS monitors percent oxygen (%O2) in air above aliquots of PCs that have been filtered to remove the confounding effect of respiring PLTs and residual WBCs.

STUDY DESIGN AND METHODS:  One‐day‐old WBC‐reduced whole‐blood‐derived PCs (WBPCs) were inoculated with bacteria at 100 to 500 CFU per mL. After 30 minutes, 2‐ to 3‐mL aliquots were processed through a PLT‐reducing filter into a sample pouch containing sodium polyanethol sulfonate and entrained air. After incubation at 35°C for at least 24 hours, the %O2 was measured within the pouch. Noninoculated WBC‐reduced WBPCs (n = 155), confirmed free of bacteria by routine culture, were tested in a like manner. Results from the latter group of WBC‐reduced WBPCs were used to distinguish contaminated from noncontaminated units.

RESULTS:  After a 24‐hour incubation at 35°C, 195 (96.5%) of the 202 sample pouches obtained from inoculated units were detected by the BDS. After an additional 6 hours at room temperature, those that remained and were tested were found positive. None of the noninoculated controls produced a positive reading.

CONCLUSION:  The BDS is easy to use and provides good levels of sensitivity and specificity.
Multicenter comparison of serologic assays and estimation of human herpesvirus 8 seroprevalence among US blood donors
Authors: Pellett P.E. , Wright D.J. , Engels E.A. , Ablashi D.V. , Dollard S.C. , Forghani B. , Glynn S.A. , Goedert J.J. , Jenkins F.J. , Lee T.‐H. , Neipel F. , Todd D.S. , Whitby D. , Nemo G.J. , Busch M.P.
Published: December 2004

BACKGROUND:  As part of assessing the possibility of transfusion transmission of human herpesvirus 8 (HHV‐8 or Kaposi's sarcoma‐associated herpesvirus), HHV‐8 seroprevalence was estimated among US blood donors, the performance of HHV‐8 serologic tests was compared, and the presence of HHV‐8 DNA was tested for in donated blood.

STUDY DESIGN AND METHODS:  Replicate panels of 1040 plasma specimens prepared from 1000 US blood donors (collected in 1994 and 1995) and 21 Kaposi's sarcoma patients were tested for antibodies to HHV‐8 in six laboratories. HHV‐8 PCR was performed on blood samples from 138 donors, including all 33 who tested seropositive in at least two laboratories and 22 who tested positive in at least one.

RESULTS:  The estimated HHV‐8 seroprevalence among US blood donors was 3.5 percent (95% CI, 1.2%‐9.8%) by a conditional dependence latent‐class model, 3.0 percent (95% CI, 2.0%‐4.6%) by a conditional independence latent‐class model, and 3.3 percent (95% CI, 2.3%‐4.6%) by use of a consensus‐derived gold standard (specimens positive in two or more laboratories); the conditional dependence model best fit the data. In this model, laboratory specificities ranged from 96.6 to 100 percent. Sensitivities ranged widely, but with overlapping 95 percent CIs. HHV‐8 DNA was detected in blood from none of 138 donors evaluated.

CONCLUSIONS:  Medical and behavioral screening does not eliminate HHV‐8‐seropositive persons from the US blood donor pool, but no viral DNA was found in donor blood. Further studies of much larger numbers of seropositive individuals will be required to more completely assess the rate of viremia and possibility of HHV‐8 transfusion transmission. Current data do not indicate a need to screen US blood donors for HHV‐8.
Comparison of two apheresis systems for the collection of CD14+ cells intended to be used in dendritic cell culture
Authors: Strasser Erwin F. , Berger Thomas G. , Weisbach Volker , Zimmermann Robert , Ringwald Jürgen , Schuler‐Thurner Beatrice , Zingsem Jürgen , Eckstein Reinhold
Published: December 2004

BACKGROUND:  Monocytes collected by leukapheresis are increasingly used for dendritic cell (DC) culture in cell factories suitable for DC vaccination in cancer.

STUDY DESIGN AND METHODS:  Using modified MNC programs on two apheresis systems (Cobe Spectra and Fresenius AS.TEC204), leukapheresis components collected from 84 patients with metastatic malignant melanoma and from 31 healthy male donors were investigated. MNCs, monocytes, RBCs, and platelets (PLTs) in donors and components were analyzed by cell counters, WBC differential counts, and flow cytometry.

RESULTS:  In 5‐L collections, Astec showed better results regarding monocyte collection rates (11.0 vs. 7.4 × 106/min, p = 0.04) and efficiencies (collection efficiency, 51.9 vs. 31.9%; p < 0.001). Both devices resulted in monocyte yields at an average of 1 × 109 (donors) and 2.5 × 109 (patients), whereas Astec components contained high residual RBCs. Compared to components with low residual PLTs, high PLT concentration resulted in higher monocyte loss (48 vs. 20%, p < 0.0001) before DC culture.

CONCLUSION:  The Astec is more efficient in 5‐L MNC collections compared to the Spectra. Components with high residual PLTs result in high MNC loss by purification procedures. Thus, optimizing MNC programs is essential to obtain components with high MNC yields and low residual cells as prerequisite for high DC yields.
Effects of resistance training during bed rest on the viscoelastic properties of tendon structures in the lower limb
Authors: Kubo Keitaro , Akima Hiroshi , Ushiyama Junichi , Tabata Izumi , Fukuoka Hideoki , Kanehisa Hiroaki , Fukunaga Tetsuo
Published: December 2004

The purpose of this study was to investigate the effects of resistance training on the tendon properties in knee extensors during 20 days of bed rest. Sixteen men were assigned to the resistance training group (BR‐Tr) or the non‐training, control group (BR‐Con). Leg‐press exercises were performed as five sets of 10 repetitions at 90% of maximum load daily for 20 days during bed rest. Before and after bed rest, the elongation of the tendon structures of the vastus lateralis muscle during isometric knee extension was determined using ultrasonography, while subjects performed ramp isometric contraction up to the voluntary maximum, followed by a ramp relaxation. The relationship between estimated muscle force (Fm) and tendon elongation (L) was fitted to a linear regression curve, the slope of which was defined as stiffness. The hysteresis was calculated as the ratio of the area within the Fm–L loop to the area beneath the load portion of the curve. The stiffness decreased significantly after bed rest for BR‐Con, but not for BR‐Tr. Similarly, the hysteresis increased significantly after bed rest for BR‐Con, but not for BR‐Tr. These results suggested that the bed rest caused the stiffness of tendon structures to decrease and their hysteresis to increase, and that leg‐press training prevents the deconditioning of the tendon structures in knee extensors.
Relationship of leg muscle strength and knee function in the early period after anterior cruciate ligament reconstruction
Authors: Morrissey M.C. , Hooper D.M. , Drechsler W.I. , Hill H.J.
Published: December 2004

The purpose of this descriptive study was to assess the relationship between knee function and the strength testing performance of various leg muscles in the early period after anterior cruciate ligament reconstruction (ACLR). Twenty‐four patients who had recent ACLR (seven females and 17 males, mean age=27 years) were tested for leg muscle strength and knee function 2 weeks after surgery. The concentric peak torques of the hip and knee extensors and knee flexors were measured at 210° s−1. In addition, the latter two muscle groups were measured at 60° s−1, and also isometrically (at a knee angle of 60°). Knee function was assessed using a three‐dimensional motion analysis system combined with a force platform. This system determined knee joint angles, torques and powers during level walking, stair ascent and stair descent. Pearson's correlation coefficients were calculated to assess the relationships between the injured knee function variables and the injured/uninjured strength ratios. The correlation coefficients ranged (in absolute value) from 0.03 to 0.81, and were generally higher for the stair tasks compared with level walking, and highest for the knee extensors (as compared with the knee flexors and hip extensors), and the strength test velocity appears much less important than the muscle group tested. These results contribute evidence indicating the importance of knee extensor strength to knee function in the early period after ACLR.
Training and performance characteristics among Norwegian International Rowers 1970–2001
Authors: Fiskerstrand Å. , Seiler K. S.
Published: December 2004

This study quantified changes in training volume, organization, and physical capacity among Norwegian rowers winning international medals between 1970 and 2001. Twenty‐eight athletes were identified (27 alive). Results of physiological testing and performance history were available for all athletes. Twenty‐one of 27 athletes responded to a detailed questionnaire regarding their training during their internationally competitive years. Maximal oxygen uptake (VO2 max) increased 12% (6.5± 0.4 vs. 5.8±0.2 L min−1) from the 1970s to the 1990s. Similarly, 6‐min ergometer rowing performance increased almost 10%. Three major changes in training characteristics were identified: (1) training at a low blood lactate (<2 mM) increased from 30 to 50 h month−1and race pace and supra‐maximal intensity training (∼8–14 mM lactate) decreased from 23 to ∼7 h month−1; (2) training volume increased by ∼20%, from 924 to 1128 h yr−1; (3) altitude training was used as a pre‐competition peaking strategy, but it is now integrated into the winter preparation program as periodic 2–3‐week altitude camps. The training organization trends are consistent with data collected on athletes from other sports, suggesting a “polarized” pattern of training organization where a high volume of low intensity training is balanced against regular application of training bouts utilizing 90%–95% of VO2 max.
The role of exercise and gender for physical self‐perceptions and importance ratings in Swedish university students
Authors: Lindwall Magnus , Hassmén Peter
Published: December 2004

The purpose of this study was to investigate how scores on the Physical Self‐Perception Profile (PSPP), including scores on the Perceived Importance Profile (PIP), were related to self‐reported exercise frequency, duration, and gender in sample of Swedish university students. A total of 164 participants completed the PSPP, PIP, and a questionnaire focusing on frequency and duration of exercise. Exercise frequency, duration, and gender predicted best the PSPP sub‐domains of Sport Competence and Physical Conditioning. Exercising more frequently, and for a longer time on each occasion was associated with higher PSPP and PIP scores. Women generally displayed lower PSPP scores than men. These results suggest that exercise professionals need to master a range of appropriate exercise strategies, since doubts concerning self‐presentation may work against establishing a regular exercise routine.