bluuebunny/crossref_metadata_2025 huggingface dataset:
Red Cell and Platelet Concentrates from Blood Collected into Half‐Strength Citrate Anticoagulant: Improved Maintenance of Red Cell 2,3‐Diphosphoglycerate in Half‐Citrate Red Cells
Authors: Farrugia A. , Douglas S. , James J. , Whyte G.
Published: March 2009

This study confirms previous work suggesting equivalent in vitro properties in blood components prepared from donations collected into half‐citrate preservative (HCPD) compared to components derived from donations collected into standard citrate‐phosphate‐dextrose (CPD) preservatives. In addition, red cell products harvested from HCPD donations showed significantly improved maintenance of pH over storage, and this was reflected in improved maintenance of intracellular 2,3‐diphosphoglycerate (2,3‐DPG). This effect was observed in whole blood and in red cells suspended in a phosphate‐containing additive solution (Tuta AAS). Collection into HCPD also improved 2,3‐DPG maintenance in red cell concentrates processed following an 18‐hour hold at 22°C. These improvements were less pronounced in red cells suspended in a non‐phosphate‐containing medium (Fenwal Adsol) in which a higher pH was maintained even in units collected in CPD. Platelets harvested from HCPD blood and suspended in plasma showed equivalent quality to platelets from standard donations. Some deterioration of platelet properties was observed when HCPD platelets were stored in a non‐citrate synthetic medium. Together with data indicating improved coagulation factor stability, these results suggest that collection into HCPD improves stored blood quality and may also allow logistical benefits in blood component preparation.
Virus Inactivation of Fresh Frozen Plasma by a Solvent Detergent Procedure: Biological Results
Authors: Piquet Y. , Janvier G. , Selosse P. , Doutremepuich C. , Jouneau J. , Nicolle G. , Platel D. , Vezon G.
Published: March 2009

In order to increase the safety of blood products, we have developed a procedure for the virus inactivation of fresh frozen plasma. Several batches have been prepared and with the first 10 batches, each of them composed of 60 litres of plasma, we have determined a set of biological parameters. Virus inactivation was realised using TnBP (1%) and Octoxynol 9 (1%). After their elimination with castor oil using chromatography on insolubilized C18 resin, glycine was added and the pH of the plasma was adjusted to 7.4. Plastic bags were aseptically filled with a mean volume of 200 ml of plasma. The mean levels of coagulation factors were all over 0.7 U/ml and their recovery from initial plasma was nearly the same as total protein except for factor VIII:C. The net loss in factor VIII:C was 16%, when including the dilution of plasma. In vivo and in vitro tests demonstrated that in the final product there were no activated factors. As in fresh frozen plasma, the protein concentration was over 50 g/l and the potassium level lower than 5 mmol/l. According to these results, virus‐inactivated plasma has the same qualities of fresh frozen plasma and could now replace it.
Prevention of Primary HLA Class I Allo‐Immunization with Leucocyte‐Poor Blood Components Produced without the Use of Platelet Filters
Authors: Ten Haaft M.A. , Van den Berg‐Loonen P.M. , Van Rhenen D.J.
Published: March 2009

In a prospective study the incidence of allo‐immunization and platelet refractoriness was investigated using a consequently leucocyte‐poor blood product regime. Twenty‐five previously non‐transfused patients with acute leukaemia (11 men, 7 women) or autologous bone marrow transplantation for Hodgkin's or non‐Hodgkin's lymphoma (2 men, 5 women) received at least 80 donor units of filtered red cells (filtration within 24 h after donation, leucocyte content 8.5+3.9 times 106/U) and/or of platelet concentrate (produced by the buffy coat method, leucocyte content: 7.8+4.2 times 106/U). A 1‐hour recovery of 20% in three consecutive transfusions, in the absence of clinical factors known to impair increment, was defined as platelet refractoriness. HLA class I antibody screening with a panel of 60 cells was performed before the first transfusion and after 80 U of blood components. Of 25 patients who entered this study, 6 patients developed platelet refractoriness after a mean of 38 units of blood components (range 26–45 U); all 6 were female with a history of multiple pregnancies. In 19 patients regarded as non‐refractory, no HLA antibodies were demonstrated (13 men, 6 women). This study, though limited in size, suggests that the use of blood products containing less than 1 times 107 leucocytes/donor unit prevents primary HLA class I immunization and platelet refractoriness.
Antibody Responses to Hepatitis C Virus by Second‐Generation Immunoassays in a Cohort of Patients with Bleeding Disorders
Authors: Hatzakis A. , Polychronaki H. , Miriagou V. , Yannitsiotis A. , Chrispeels J. , Troonen H. , Karafoulidou A. , Gialeraki A. , Katsouyanni K. , Mandalaki T.
Published: March 2009

The antibody responses and the prevalence patterns of antibodies to hepatitis C virus (anti‐HCV) in a cohort of patients (n = 210) with bleeding disorders were studied using a first‐generation and a second‐generation enzyme immunoassays (EIA‐1, EIA‐2) as well as a second‐generation recombinant immunoblot assay (RIBA‐2). The anti‐HCV prevalence as determined by EIA‐1 and EIA‐2 was 183/210 (87.1%) and 197/210 (93.8%), respectively (p = 0.0026). None of the 17 EIA‐2(+)/EIA‐1(‐) samples was scored nonreactive by RIBA‐2. At follow‐up, samples of 123 patients were tested. Twenty‐nine out of 111 patients reactive by EIA‐1 seroreverted according to EIA‐1 while the sero‐reversion rate with EIA‐2 was 0 out of the 121 (p<10‐8). The anti‐HCV prevalence by EIA‐2 was 150/154 (97.4%) in anti‐HIV‐1‐positive individuals and 47/56 (83.9%) in the anti‐HIV‐1‐negative ones (p = 0.001). However, high assay signals (OD 492 nm >2.0) were observed in 94/150 (62.7%) and 45/47 (95.7%) of the anti‐HIV‐1‐positive and ‐negative patients, respectively (p = 10‐5). The decreasing anti‐HCV reactivity among anti‐HIV‐1‐positive individuals was mainly due to diminishing c33c reactivity. Seroconversion to anti‐HCV was observed in 3/7 (42.9%) cases with acute icteric non‐A, non‐B hepatitis by both EIA‐1 and EIA‐2, while the remaining 4 cases had detectable levels of anti‐HCV 1–18 months before the acute episode.
Evaluation of Monoclonal Antibodies with Specificity for the Human Platelet HPA‐1 Allotypes
Authors: Ryckewaert Jean Jacques , Marguerie Gérard , Masson Dominique , Bensa Jean‐Claude , Diaz Marline
Published: March 2009

Two mouse monoclonal antibodies have been characterized. Both antibodies specifically recognized the amino acid sequences which are responsible for the presence of the HPA‐1a and HPA‐1b alloantigens at the surface of human platelet. The monoclonality allied with an exquisite specificity of these antibodies allow a perfect typing of platelet HPA‐1 status. A fast and reliable Elisa assay which fulfills all the needs and requirements of hematology laboratories has been set up and is described here. The Elisa assay has been tested on a population of 689 blood donors. The results agreed with the known phenotypic frequencies of the HPA‐1 alloantigens.
Cold Autoagglutinins with Anti‐Pr Specificity Associated with Fresh Varicella Infection
Authors: Herron B. , Orson G. , Myint H. , Boulton F. E. , Roelcke D.
Published: March 2009

Cold agglutinins with the rare anti‐Pr specificity were identified in an adult patient with fresh varicella infection. The antibody was of the IgM ϰ type with subspecificity anti‐Pr3d and caused a haemolytic episode in the patient. In the only previously reported case in which cold agglutinins were associated with varicella infection, the antibody specificity was also anti‐Pr.
Half‐Strength Citrate CPD Combined with a New Additive Solution for Improved Storage of Red Blood Cells Suitable for Clinical Use
Authors: Högman C. F. , Eriksson L. , Cong J. , Högman A. B. , Vikholm K. , Debrauwere J. , Payrat J. M. , Stewart M.
Published: March 2009

Currently used systems for red blood cell (RBC) collection and storage for transfusion have the disadvantage that the RBC 2,3‐bisphosphoglycerate (BPG) concentration is depleted within two weeks of storage, resulting in a left‐shift of the oxygen dissociation curve and a temporarily impaired capacity to deliver oxygen. We have studied the effects on red cell metabolism, morphology and in vivo recovery of 49‐day storage of RBC, with collection in half‐strength citrate CPD (0.5CPD) and storage in an additive solution containing citrate, adenine, mannitol, phosphate and glucose (RAS2). Traditional CPD‐SAGM was used for comparison. Component preparation was performed after an initial holding period of the whole blood at ambient temperature for 8 h. The BPG concentration in 0.5CPD‐RAS2 RBC was 0.633 ± 0.120mol (mol Hb)‐1 as compared to 0.454±0.138mol (mol Hb)‐1 in CPD‐SAGM RBC which implied a decrease to 67 and 48% of normal concentration, respectively. The mean RBC BPG concentration was maintained at the initial level for 28 days in the new system but decreased to very low levels within 14 days in the controls. The total adenine nucleotides were well maintained in both systems, adenosine triphosphate slightly better in the new system. Hemolysis after 49 days was 0.35±0.21% in the new system and 0.72 ±0.25% in the controls (p<0.001). The morphology was better maintained in the new system (p<0.001). The 24‐hour posttransfusion survival of 49‐day stored RBC was 78.9±7.1%. The membrane leakage of sodium and potassium was not significantly different in the two systems. The concentration of inorganic phosphate (Pi) in the extracellular fluid of 0.5CPD‐RAS2 RBC was initially 7.1±0.8 mmoll‐1. It decreased to approximately 5.5 mmol l‐1 during the first 2 weeks of storage and then slowly increased to 9.4±0.9 mmol l‐1 on day 49. Pi in the CPD‐SAGM controls was low initially but rose to 6.2±0.4 mmol l‐1 at the end of storage. The new system offers clearcut advantages as compared to traditional systems and does not seem to raise any serious clinical objections against its use even in massive transfusions.
A Simplified Assay for the Specific Diagnosis of Paroxysmal Nocturnal Hemoglobinuria: Detection of DAF(CD55)‐ and HRF20(CD59)‐ Erythrocytes in Microtyping Cards
Authors: Nilsson Bo , Hagström Ulla , Englund Åsa , Säfwenberg Jan
Published: March 2009

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare disease that is caused by a monoclonal stem cell defect. The affected cells lack the carbohydrate linkage between phophatidylinositol and a group of membrane proteins of which three protect the cell against complement lysis. The absence of these three proteins, DAF (CD55), C8BP and HRF 20 (CD59), makes cells from the erythropoiesis, thrombopoiesis and myelopoiesis extensively sensitive to complement attack and affected patients suffer from intravascular hemolysis, thrombosis and increased susceptibility to infections. In this study we describe a swift and specific assay for the detection of CD55‐ and CD59‐ erythrocytes, which is suitable for screening of possible PNH patients.
Intravenous Immunoglobulin Therapy of Autoimmune and Systemic Inflammatory Diseases
Authors: Ronda Nicoletta , Hurez Vincent , Kazatchkine Michel D.
Published: March 2009

The proven beneficial effect of intravenous immunoglobulins (IVIg) in certain autoimmune disorders has led to the development of clinical trials in other autoimmune and systemic inflammatory diseases. In parallel, experimental studies are being carried out to better understand the mechanisms of action of IVIg. In this review, we discuss the clinical use of IVIg in autoimmune disorders and the possible mechanisms by which IVIg may be effective in the various diseases. A better understanding of the mechanisms of action of IVIg in autoimmune disease will allow optimization of their use as a therapeutic alternative to conventional immunosuppression.
Blood Transfusion and Immunomodulation: A Possible Mechanism
Authors: Mincheff Milcho S. , Meryman Harold T. , Kapoor Veena , Alsop Patricia , Wötzel Michael
Published: March 2009

To induce an immunogenic response in vivo, an antigen‐presenting (stimulator) cell must present both antigen‐specific (class II MHC) and an accessory signal to the CD4 T cell. Failure to express the accessory signal has been shown in vitro to induce a state of specific unresponsiveness (anergy) in the T cell. We have shown that although stimulator cells in blood continue to express class II MHC molecules during refrigerated storage, their ability to present the accessory signal diminishes, reaching zero in all units tested by about 13 days. This implies that blood in excess of 2 weeks old should not alloimmunize but could induce some degree of immunosuppression. UV‐B irradiation and, to a lesser extent, gamma‐irradiation, were also shown to inhibit expression of the accessory signal by stimulator cells in blood.
Use of Anti‐GOR Testing in the Screening of Blood Donors for Hepatitis C Virus Infection
Authors: Irving William L. , Day Sarinder , Bennett Diane , Eglin Roger P. , Flanagan Peter , Nuttall Philip , James Virge
Published: March 2009

Sera from 12/1,155 (1%) anti‐HCV‐negative (Elisa) UK blood donors were found to be anti‐GOR positive. None out of 12 of those sera were positive for HCV RNA by the reverse transcriptase/polymerase chain reaction (RT/PCR). In a cohort of 316 anti‐HCV Elisa‐positive sera, 27/57 RIBA‐positive, and 1/188 RIBA‐negative sera were anti‐GOR positive, resulting in a sensitivity of 47% and a specificity of 99.5% for anti‐GOR as a marker for RIBA‐confirmed HCV infection. Four out of 71 (6%) of RIBA‐indeterminate sera were anti‐GOR positive. Donors with anti‐GOR reactivity were more likely to have antibodies against each of the individual HCV antigens represented in the RIBA, and those antibodies were of greater intensity, when compared to the anti‐GOR negative cohort. Twenty out of 36 (55.6%) of RT/PCR‐positive sera were anti‐GOR positive, compared to 1/7 (14.3%) of RT/PCR‐negative sera (p = 0.09). The usefulness of anti‐GOR testing of UK blood donors is discussed in the light of these results.
Human Monoclonal Anti‐D‐Induced Clearance of Human D‐Positive Red Cells in a Chimpanzee Model
Authors: Blancher A. , Socha W. W. , Roubinet F. , Rowe A. W. , Broly H. , Byrne P. , Holuigue M. , Bouzidi A. , Huart J. J. , Ruffié J.
Published: March 2009

Two human monoclonal anti‐Rh0(D) antibodies, one IgG1, and one IgG3, were tested for their ability to clear human D‐positive red blood cells (RBCs) from chimpanzee circulation. Human RBCs (phenotype A1, R1r) from 1 donor were radiolabelled with chromium 51 and injected into 4 chimpanzees. One day later the control animal received isotonic saline whereas 2 animals received 400 μg of purified human monoclonal anti‐D, either IgG1 or IgG3. The remaining animal received both antibodies together (200 μg of IgG1 and 200 μg of IgG3). Both individual antibody‐mediated clearance of human D‐positive cells and synergy was not observed when both antibodies were used in combination. IgG1 was slightly more effective than IgG3. This animal model is a suitable alternative for conducting in vivo experiments in human beings, especially at the preclinical study phase of monoclonal anti‐D antibodies.
Platelet Concentrates in an Additive Solution Prepared from Pooled Buffy Coats
Authors: Eriksson Lars , Shanwell Agneta , Gulliksson Hans , Högman Claes F. , Svensson Leif A. , Kristensen Jörgen , Berg Barbro
Published: March 2009

Leukocyte‐depleted platelet concentrates were prepared from pools of 4 buffy coats on the day after blood collection (BC‐PC). The storage medium was composed of citrate phosphate dextrose plasma and a platelet‐additive solution. Autologous transfusions of 111In‐labelled platelets in 9 healthy volunteers were performed on the day of preparation (day 1) and on day 5. The recovery was 54.6±8.7 (day 1) and 51.9±10.4% (day 5), T1/2 was 101±28 and 61±9 h, respectively. The survival was 8.3±1.7 and 5.7±1.0 days, respectively, using linear plot, and 7.8±2.0 and 5.8±0.5 days using the multiple hit method. In a prospective clinical study a comparison of the corrected posttransfusion increments was made between BC‐PCs and apheresis‐PCs, and between fresh (1–2 days) and stored (3–5 days) preparations. No difference was found between BC‐PCs and apheresis PCs. However, fresh BC‐PCs gave higher increments than stored BC‐PCs. A slight numerical difference between fresh and stored apheresis‐PCs was not statistically significant. It is concluded that the BC‐PC method results in platelets of equal quality to apheresis‐PC.
Rheological Evaluation of X‐Ray Irradiated Blood
Authors: Suzuki Yoji , Seiyama Akitoshi , Tateishi Norihiko , Yamanishi Shigeki , Maeda Nobuji
Published: March 2009

CPD blood, in glass containers and polyvinyl chloride (PVC) bags, was X‐irradiated with 50 Gy (n = 4), and then stored for up to 4 weeks. Rheological, hematological and biochemical changes of the red cells during storage were examined. Changes in blood viscosity and various properties of red cells (deformability, shape, hematological indices and concentrations of 2,3‐diphosphoglycerate and nucleotides) during storage at 4°C were not altered by the irradiation. The rheological, biochemical and morphological properties of red cells were as well preserved in PVC bags as in glass containers. No abnormal and/or cross‐linked proteins in red‐cell membranes were induced by the irradiation. Irradiation of PVC bags did not affect the above properties of red cells through any changes in the bag constituents. In conclusion, X‐ray irradiation with 50 Gy to CPD blood in PVC bags does not affect the rheological functions of red cells.
Taiwan Blood Donation Program: A Model for Development
Authors: Tsai S. J. L. , Lee T. D.
Published: March 2009

In 1974, the Taiwan National Blood Program had not yet been fully developed. The volunteer donor base was less than 5% and there was no organized national structure or regulation of blood services. Following the founding of the Blood Donation Association and the Blood Services Foundation, promotion of an all‐volunteer blood program, increased utilization of blood components and the standardization of laboratory methods were strategies that led to the development of the Taiwan National Blood Program. In 1991, 1,066,082 units of blood were collected in Taiwan, serving a population of about 20 million. There are currently 6 major blood centers and 13 blood stations. Blood donation is now 100% volunteer and 78% of all blood collected is separated into 10 available components. With national organization and regulation, the transition has taken 17 years and has resulted in a community‐based, all‐volunteer blood supply.
Sensitivity of the Fibrinogen Clotting Time: An in vitro Test of Potential Thrombogenicity
Authors: Feldman P.A. , McGrath S. , Evans H.
Published: March 2009

The fibrinogen clotting time (FCT) is a measure of thrombin activity, and is used to evaluate the potential thrombogenicity of prothrombin complex concentrates (PCC). We have defined end points for clot formation in this test which allow the measurement in PCC of thrombin concentrations as low as 0.001 IU/ml. The FCT of thrombin and PCC samples which did not contain antithrombin III (ATIII) were the same when measured at 20°C or 37°C. In the presence of ATIII (0.05 or 0.25 IU/ml), samples of PCC which were known to contain thrombin showed shorter FCT at 20°C than at 37°C. Inclusion of both ATIII (0.25 IU/ml) and heparin (4 IU/ml) in PCC ensured the complete inactivation of endogenous thrombin.
Second‐Generation Anti‐HCV Screening in a Saudi Arabian Donor Population
Authors: Bernvil S. S. , Andrews V.J. , Sasich F.
Published: March 2009

The 2nd‐generation anti‐HCV test system was applied to a Saudi Arabian multi‐ethnic donor population. When donors were stratified according to first‐time donations versus repeat donations, the latter having been screened previously by a 1st‐generation set of tests, it was found that in Saudi Arabian and Middle East nationals, the 2nd‐generation tests (EIA and RIBA), identified close to double the number of anti‐HCV‐positive donors, compared to an earlier study using the 1st‐generation tests. Part of this finding was due to a 38% higher rate of RIBA‐confirmable repeat‐positive EIA results. In groups of donors, previously screened by the 1st‐generation system, some additional cases of anti‐HCV reactivity were identified, most prominently in Middle East nationals. It is assumed that some of these represented recent seroconversions, while others were cases of serologic subtypes of HCV, not reacting in the 1st‐generation tests. The current test system identifies 0.66% of Saudi‐Arabian, and 2.87% of other Middle East donors as putative carriers of hepatitis C virus. The study lends support to the opinion that donors who return regularly over the years have a lower prevalence of disease markers, thereby being a safer source of blood than first‐time donors.
Isotypes and IgG Subclasses of Anti‐Fab Antibodies in Human Immunodeficiency Virus‐Infected Hemophilia Patients
Authors: Süsal Caner , Oberg Hans‐Heinrich , Daniel Volker , Dörr Colette , Terness Peter , Huth‐Kühne Angela , Zimmermann Rainer , Opelz Gerhard
Published: March 2009

We reported recently that anti‐Fab autoantibodies of the IgG isotype are associated with the decrease of helper/inducer (CD4+) lymphocytes in human immunodeficiency virus‐infected (HIV+) hemophilia patients with acquired immunodeficiency syndrome (AIDS) or AIDS‐related complex (ARC). In the present study we investigated the subclass distribution of IgG‐anti‐Fab autoantibodies, and whether anti‐Fab antibodies of the IgA and IgM isotypes also are associated with the development of AIDS. Sera of HIV+ patients with AIDS had significantly higher IgA‐anti‐Fab activity than HIV+ patients with ARC (p<0.02), HIV+ patients without AIDS/ARC (p<0.0001), HIV‐negative (HIV‐) patients (p<0.001), or healthy controls (p<0.0001). An inverse association was found between IgA‐anti‐Fab activity and CD4+ cell counts (r = ‐0.396, p<10‐6). In contrast, no association of CD4+ cell counts was observed with IgM‐anti‐Fab. However, IgM‐anti‐Fab was significantly increased in patients with thrombocytopenia. We found a significant association between IgA‐anti‐Fab activity and serum neopterin concentrations (r = 0.310, p<10‐5). IgG‐anti‐Fab activity was detected mainly in the IgG3 fraction, although in HIV+ patients with AIDS/ARC various IgG subclasses were present. Affinity‐purified anti‐Fab antibodies isolated from sera of AIDS patients bound to rgp120‐preincubated CD4+ cells of a healthy individual, supporting our hypothesis that anti‐Fab antibodies and free circulating gp120 molecules are involved in the elimination of uninfected CD4+ cells. Removal of anti‐Fab autoantibodies from the circulation by immune adsorbance might be a useful approach in the treatment of AIDS.
Alloimmunisation via Previous Transfusion Places Female Kpb‐Negative Recipients at Risk for Having Children with Clinically Significant Hemolytic Disease of the Newborn
Authors: Gorlin Jed B. , Kelly Lydia
Published: March 2009

We report a case of clinically significant hemolytic disease of the newborn due to Kpb alloimmunisation requiring obstetric intervention. This case and a review of the literature are in contrast to reviews of hemolytic disease of the newborn that either ascribe no significance to the Kpb antigen or suggest that it causes only rare or mild disease. Analysis of our Kpb‐negative donor pool suggests that prior transfusion greatly increases the chance of alloimmunisation. The role of frozen rare donor registry cells as a public resource is emphasised.
Pure Anti‐Doa Stimulated by Pregnancy
Authors: Roxby D.J. , Paris J.M. , Stern D.A. , Young S.G.
Published: March 2009

Pure anti‐Doa stimulated by pregnancy was detected in 2 non‐transfused females during routine antenatal screening. Anti‐Doa occurs rarely and has generally been reported in combination with other antibodies. The first, and only report to date, of pure anti‐Doa was also stimulated by pregancy. We believe these instances to be only the second and third reported cases of pure anti‐Doa.
A Human Monoclonal Antibody to High‐Frequency Red Cell Antigen Jra
Authors: Miyazaki T. , Kwon K. W. , Yamamoto K. , Tone Y. , Ihara H. , Kato T. , Ikeda H. , Sekiguchi S.
Published: March 2009

A human‐mouse heterohybridoma (HMR0921) secreting human monoclonal IgG3, Λ antibody was produced from peripheral blood lymphocytes of a healthy blood donor with serum antibody to Jra, by EBV transformation and hybridization with mouse myeloma cell line P3X63Ag8.653. The reactivity of HMR0921 antibody was assessed by antiglobulin test with a panel of red cells including 14 different rare blood types. Only Jr(a‐) red cells were negative. The strict specificity of this antibody to Jra antigen was further confirmed by absorption test with fluorescence flow cytometry. On screening of 28,744 blood donor samples by HMR0921 antibody, we detected 19 agglutination‐negative samples, which were confirmed as Jr(a‐) by conventional anti‐Jra antisera. Therefore, our HMR0921 antibody is extremely useful for detecting rare Jr(a‐) blood.
Production of a New Murine Monoclonal Antibody with Fy6 Specificity and Characterization of the Immunopurified N‐Glycosylated Duffy‐Active Molecule
Authors: Riwom S. , Janvier D. , Navenot J. M. , Benbunan M. , Muller J. Y. , Blanchard D.
Published: March 2009

A murine monoclonal antibody (mAb i3A; IgG1, kappa light chain) was obtained using human red blood cells as immunogen. The antibody showed Fy6 specificity since it agglutinated all but Fy(a‐b‐)‐untreated red cells and failed to agglutinate chymotrypsin‐treated cells. An erythrocyte membrane protein of 42–46 kD was revealed as the major component recognized by the antibody on immunoblots. The antibody also bound to 92‐ to 95‐ and 200‐kD proteins, tentatively identified as oligomers of the 42‐ to 46‐kD monomeric form. The affinity‐purified Fy6‐active protein was converted to a sharp band of 35 kD after N‐glycanase treatment. The molecule appeared as a slightly broadly band after neuraminidase treatment but was not further altered by O‐glycanase. The i3A mAb bound to 6,000±1,000 receptor sites on either Fy(a‐b+), Fy (a+b+) and Fy(a+b‐) red cells with an affinity constant in the range of 3–6 times 108 M‐1. No binding was observed to other blood cells nor to several cells (B, T, myelomonocytic and erythro‐leukemia cell lines). Also, the bulk of i3A‐Fy6 immune complexes could be dissociated from the red cell membrane with as low as 0.2% Triton X‐100, showing that the Fy6‐active glycoprotein is not tightly associated with the membrane skeleton. Our data obtained with a new monoclonal antibody directed to the Fy6 antigen demonstrate that the blood group Duffy‐active component is a red cell‐specific glycoprotein carrying one or more N‐linked oligosaccharides.
Determinants of Physician Ordering of Preoperative Autologous Donations
Authors: AuBuchon J. P. , Gettinger A. , Littenberg B.
Published: March 2009

Preoperative autologous donation (PAD) is frequently utilized to reduce infectious disease transmission risks, but it is an expensive form of hemotherapy that is not always closely matched with the needs of the patients. We compared the use of PAD in an academic medical center with patient needs and then investigated the efficacy of a simple intervention to promote targeting of PAD toward patients most likely to benefit from having PADs available. Over a 3‐month period, surgeons whose patients received allogeneic components were asked to complete a questionnaire designed to identify reasons for not ordering PAD. PAD units were used in 14 (11%) of 124 cases, accounting for 6% of perioperative red cell use. The responses (46, 42% of the surveys sent) stated that PAD had not been ordered because of time constraints (39%), medical problems (26%), anemia (15%), and lack of expectation of blood use (24%). Chart review documented the presence of cited conditions in 88% of cases. Logistic and cost concerns were not evident. However, only 8% of the 176 PAD units collected in that period were transfused. Following dissemination of PAD ordering guidelines, this proportion rose to 52% without a reduction in the proportion of elective surgical cases utilizing autologous transfusion. This improvement in ordering practice was maintained over at least in 5‐month period. Thus we were able to improve the efficiency of PAD application (reducing over‐ordering) through a simple feedback to surgeons that assisted them in targeting PAD toward patients most likely to need transfusion.
Expression of A Antigens on Erythrocytes of Weak Blood Group A Subgroups
Authors: Heier Hans Erik , Namork Ellen , Čalkovská Zdenká , Sandin Randi , Kornstad Leif
Published: March 2009

Scanning immune electron microscopy using a monoclonal anti‐A antibody which reacts with all type A oligosaccharide chains revealed A antigens on less than 5% of Am and Ael cells, some of which showed extremely strong labelling. This explains why Am and Ael cells can absorb significant amounts of anti‐A without being agglutinated. A3 may be a heterogenous subgroup, since A antigens were found on 82 and 58%, respectively, of the cells of 2 A3 individuals. A antigens were found on 75% or more of Ax cells. In many weak A individuals A‐positive cells are apparently best detected if an anti‐A is used which reacts strongly with other A oligosaccharide chains than type 2. From hyperimmune pregnancy sera Ax, Am and Ael erythrocytes absorbed antibodies which seemed to have other fine specificities than those absorbed by A2 cells. We conclude that weak subgroups of A may deviate from A2 both by number of erythrocytes expressing A antigens and the biochemical nature of the antigens.
In vitro Evaluation of a New Filter for Leucocyte Depletion of Platelet Concentrate during Component Preparation
Authors: Christensen Lisa Dalh , Dickmeiss Ebbe
Published: March 2009

A newly developed filter for prestorage leucocyte depletion of platelet concentrates (PC) was studied. The filter is designed for leucocyte depletion during the preparation of the pool PC from platelet rich buffy coats. In all the leucocyte depleted PC (LD‐PC) leucocyte depletion was satisfactory. 19 of 20 units of LD‐PC had a leucocyte content below 3times105per PC and 1 contained 8times105leucocytes. The standard PC contained 2.53times108(0.87times108–15.3times108; n = 20) leucocytes per PC (median and range). The quality of the LD‐PC was evaluated by measuring platelet activation, platelet morphology, and pre‐ and poststorage pH. There were no differences in any of the parameters studied.
Anti‐HCV Seroconversion in Multitransfused and Immunocompromised Patients A Long‐Term Longitudinal Study
Authors: Cornu Chantal , Brusselmans Caroline , Geubel André , Ferrant Augustin , Cornu Guy
Published: March 2009

In a series of 6 multitransfused, immunocompromised patients, the diagnosis of posttransfusion hepatitis C was based upon the analysis of long‐term follow‐up serum samples. The HCV RNA was detected by a nested PCR assay using primers located in the 5′ noncoding region (5′NCR), and anti‐HCV antibodies were assayed with third‐generation tests. The interval between the first rise in alanine aminotransferase and seroconversion varied from less than 5 months to more than 38 months. Five out of 6 patients seroconverted after 14 months or later. In most cases, the anti‐NS3 and anti‐NS4 antibodies appeared first. In such patients, the etiology of chronic liver disease may thus be overlooked for 1 or more years, a definitive diagnosis requiring the detection of HCV RNA.
Molecular Bases of the ABO Blood Groups of Indians from the Brazilian Amazon Region
Authors: Franco Rendrik F. , Simôes Belinda P. , Guerreiro Joāo F. , Santos Sidney E.B. , Zago Marco A.
Published: March 2009

Phenotype studies of ABO blood groups in most Amerindian populations revealed the exclusive presence of group O. Since group O is the result of the absence of glycosyltransferase activity, its molecular bases may be heterogeneous. We carried out ABO blood group genotyping by analysis of DNA of 30 Indians from 2 Amazonian tribes (Yanomami and Arara), and compared the findings with other populations (Caucasians and Blacks). Two segments of the glycosyltransferase gene were amplified by PCR and digested with KpnI or AluI to detect deletion or base change at positions 258 and 700, respectively. For all subjects, the gene basis of blood group O is the deletion of a single nucleotide at position 258 of the glycosyltransferase A gene, similar to that observed in Caucasoids and Negroids. DNA sequencing of limited regions of the gene supports this conclusion. This finding does not exclude, however, that a heterogeneity of the O allele may be revealed by a more extensive analysis.
Production of Endothelin‐1 Induced by Blood Transfusion in Premature Infants
Authors: Sasai‐Takedatsu Misa , Kojima Takatsugu , Hirata Yukio , Kobayashi Yohnosuke
Published: March 2009

We demonstrated a relationship between blood transfusion and plasma ET‐1 levels in 10 premature infants with anemia. Changes in plasma ET‐1 and lipoperoxide levels, blood pressure and available oxygen before and after the blood transfusion were determined. Plasma ET‐1 levels were significantly elevated after blood transfusion (before: 8.0+2.8; after: 15.7+5.4 pg/ml, p<0.01). A positive correlation was observed between plasma ET‐1 levels and lipoperoxide levels (r = 0.887, p<0.01). There was a significant correlation between plasma ET‐1 levels and transfused blood volumes (r = 0.758, p<0.01). No positive correlation was observed between plasma ET‐1 levels and available oxygen. These results suggest that blood transfusion may be a stimulator of ET‐1 overproduction and that it may lead to tissue injury through an endothelin‐induced oxygen radical formation system.
Blood Safety in Developing Countries
Authors: Gibbs William N. , Corcoran Patricia
Published: March 2009

This study, based on responses to a questionnaire, was undertaken to define problems in and formulate solutions for improving blood safety in developing countries as part of an effort to monitor the status of blood transfusion services globally. Despite improvements between 1988 and 1992, only 66% of developing countries (DGCs) and 46% of least developed countries (LDCs) screen all blood donations for antibodies to human immunodeficiency viruses; 72% DGCs and 35% LDCs test all donations for hepatitis B surface antigen and 71 and 48%, respectively, for syphilis. The antihuman globulin test is performed routinely in 62% DGCs and 23% LDCs, and inadequate quality assurance in all aspects of preparatory testing is a major weakness in many countries. The blood supply is usually insufficient: none of the LDCs and 9% of the DGCs collect 30 units or more per 1,000 of the population annually. Blood donor systems are totally voluntary and non‐remunerated in 15% DGCs and 7% LDCs; 80% DGCs and 93% LDCs rely totally or partially on replacement donors and 25% of both groups on paid donations. The proportion of repeat donors is low (medians: 47% in DGCs, 20% in LDCs), and discard rates for collected blood are often high (up to 33%). Most of the blood collected is transfused as whole blood, and most DGCs and LDCs have inadequate supplies of plasma substitutes for management of acute haemorrhage. The reasons for these problems and suggested solutions are discussed.
Flow Cytometry: A Tool in Immunohematology for D+w(Du) Antigen Evaluation?
Authors: Tazzari Pier Luigi , Bontadini Andrea , Belletti Daniela , Malferrari Franco , Conte Roberto
Published: March 2009

We performed a flow‐cytometric analysis of the expression of D antigen in D+w samples (previously termed Du). We also analysed a series of D‐positive and D‐negative (cde phenotype) samples to obtain positive and negative controls, respectively. The evaluation was carried out by immunofluorescence and the intensity of positivity was expressed as mean channel value (MCV) of fluorescence. Results demonstrated that D+w samples have lower expression (less than 1 log) than D‐positive cases (p>0.001, Student t test), while cde samples show the same MCVs as negative controls. Moreover, it was also possible to set a grading of D antigen expression and to analyze cases difficult to assess by agglutination only.
Donor Follow‐Up of Influenza Vaccine‐Related Multiple Viral Enzyme Immunoassay Reactivity
Authors: Arnold N.L , Slade B.A. , Jones M.M. , Popovsky M.A.
Published: March 2009

False‐positive enzyme immunoassay (EIA) tests in blood donors receiving influenza vaccine were first reported in 1991. We conducted follow‐up testing for 6 months of those donors with multiply reactive, but unconfirmed EIA (at least 2 positives in anti‐HCV‐1.0, anti‐HIV‐1, and anti‐HTLV‐I assays) with a history of recent flu vaccine to determine the duration of false positivity. Of 133,000 donors tested, 16 met study criteria; all 16 were reactive for anti‐HCV, 10 were reactive for anti‐HIV‐1, and 12 were reactive for anti‐HTLV‐I. Fifteen donors were available for follow‐up testing (using the original screening and supplemental tests): 10 (67%) reverted to negative for the 3 tests and 5 remained false positive for various markers at last sampling (3–6 months after vaccination). The mean duration of false positivity for those reverting to negative EIA test status, was 4.2 months (range 2–7 months) indicating a transient phenomenon and supporting studies which suggest a role for IgM in the mechanism.
Effectiveness of High‐Dose Intravenous Gamma Globulin Therapy in Acquired von Willebrand's Disease
Authors: van Genderen Perry J.J. , Michiels Jan J. , Bakker Jet J. , van't Veer Mars B.
Published: March 2009

An 82‐year‐old patient with acquired von Willebrand's disease in association with a non‐Hodgkin lymphoma and a benign IgGλ monoclonal paraproteinaemia is described with severe recurring nasopharyngeal bleedings, who responded poorly to desmopressin (DDAVP) and factor VIII/von Willebrand factor concentrates but was successfully treated with high‐dose intravenous γ‐globulin therapy.
Microbiologic Contamination of Peripheral Blood Stem Cell Autografts
Authors: Schwella N. , Zimmermann R. , Heuft H.G. , Blasczyk R. , Beyer J. , Rick O. , Kreißig C. , Zingsem J. , Eckstein R. , Siegert W.
Published: March 2009

We have determined the incidence and clinical significance of positive microbiologic cultures in a series of 290 peripheral blood stem cell concentrates in 95 patients undergoing multiple apheresis procedures for autologous stem cell rescue. Specimens for bacterial cultures were obtained after processing of the autografts just prior to freezing. The incidence of microbial contamination was 4.5% (n = 13). The predominant pathogenic microorganism cultured was coagulasenegative Staphylococcus (n = 11). From 8 patients with contaminated leukapheresis products 6 underwent autologous stem cell transplantation. Five patients received 1–5 culture‐positive stem cell concentrates without serious sequelae, whereas the sixth patient was autografted with noncontaminated leukapheresis products, 1 concentrate contaminated with Aspergillus fumigatus being not reinfused. No microorganism present in the stem cell autograft was recovered in vivo in the posttransplantation period, although fever as a sign of infection occurred in all but 1 patient. Peripheral blood stem cell collection and ex vivo processing for cryopreservation may result in microbiologic contamination. However, our data show that infusion of contaminated stem cell autografts does not play a significant role as a source for infections in the clinical setting of autologous stem cell rescue.
An Analysis of Autologous Blood Donor Motivational Factors
Authors: Domen R.E. , Ribicki L.A. , Hoeltge G.A.
Published: March 2009

Little is known about the patient's reasons for participating in an autologous blood (AUB) collection program. We surveyed 110 AUB donors in our hospital‐based collection program. Although fear of infection from an allogeneic blood transfusion was cited by 20% of AUB donors, 68.2% indicated that their physician's recommendation was a motivating factor. For 18.2% self‐initiated motivation was a factor in pursuing AUB. The overwhelming majority (97.1%) stated they would donate AUB even if the risk of getting AIDS from a blood transfusion was zero. Fear of infection does not appear to be the primary reason for patients to donate AUB. Recommendation by their personal physician is an important component in the patients' decision‐making process to set aside their own blood for upcoming surgery.
Is Allergy Screening of Blood Donors Necessary?: A Comparison between Questionnaire Answers and the Presence of Circulating IgE Antibodies
Authors: Stern Annika , van Hage‐Hamsten Marianne , Sondell Kerstin , Johansson S. G. O.
Published: March 2009

We investigated 477 consecutive blood donors in order to find the predictive value of a questionnaire as a screening method for preventing passive transfer of IgE antibodies to common allergens that might cause transfusion reactions. Of the 477 donors, 119 (24.9%) claimed to be allergic and 358 (75.1%) recorded no allergic complaints. Serological examination with Phadiatop® and a multi‐allergen mix for food (fx5) detected IgE antibodies in only 54.5% of the allegedly alllergic blood donors. Among the allegedly asymptomatic donors, 12.3% showed positive results in these tests. Subsequently, radioallergosorbent test analyses with common inhalant and food allergens were performed to specify the IgE responses obtained. Our results demonstrate a remarkable difference between the questionnaire answers and the serological measurements of IgE antibodies and raise the question of whether – and if so, what kind of – allergy screening is warranted among presumptive blood donors.
A Quantitative Assay for Subclassing IgG Alloantibodies Implicated in Hemolytic Disease of the Newborn
Authors: Thomas Nanette C. , Shirey Rosetta S. , Blakemore Karin , Kickler Thomas S.
Published: March 2009

Traditionally, IgG subclassing has been performed using qualitative assays. Quantitation of IgG subclasses may have prognostic value in evaluating alloimmunized pregnancies. A quantitative enzyme‐linked immunosorbent assay (ELISA) was implemented for measuring IgG subclasses of red blood cell (RBC) antibodies (AB) isolated by adsorption/elution from the sera of alloimmunized pregnant women. The assay is a sandwich enzyme immunoassay using monoclonal antibodies specific for the relevant IgG subclasses and anti‐human IgG peroxidase conjugate to quantitate the amount of bound IgG. The sensitivities of the assay for IgG1, 2, 3, and 4, respectively, were 4,23,4 and 2 μg/l. The results for each subclass for a given AB were expressed as a percentage of the total. In a series of pregnant mothers with ABs: E (4), Fya (2), Jka (1) and S (1), the mean percentage ± 1SD of each subclass was: IgG1 61±34; IgG2 14±22; IgG3 18±28 and IgG4 4±17. IgG1 or IgG3 accounted for greater than 50% of the AB subclass distribution in 5 cases that resulted in hemolytic disease of the newborn (HDN). Although only a small number of samples was studied, changes in the concentrations of IgG1 or IgG3 during gestation suggest a correlation with the presence or absence of HDN. The ELISA may be used to quantitate the IgG subclasses of RBC ABs and may be valuable in predicting the severity of HDN in alloimmunized pregnancies.
Epidemiology of Inhibitors in Haemophilia A
Authors: Kreuz W. , Escuriola‐Ettingshausen C. , Martinez‐Saguer I. , Güngör T. , Kornhuber B.
Published: March 2009

One of the most serious complications of the treatment of haemophilia A is the development of inhibitors. Former studies mostly considered the prevalence of inhibitor development, thus underestimating its true risk. Prevalences ranged widely (7–18%) probably due to the populations studied and the study design. Recent prospective previously untreated patients (PUP) studies were more comparable because of similar study designs. Eight PUP studies regarding the incidence of factor VIII inhibitors were analyzed: The inhibitor incidences (Independent of severity of haemophilia) ranged from 18.4 to 28%. Evaluating only severe haemophiliacs (factor VIII<2%) significantly higher incidences were found. After 9–36 exposure days (as medians inhibitor development occurred at 0.8‐3.3 years of age (as medians).
Butyrylcholinesterase: biomarker for exposure to organophosphorus insecticides
Authors: Stefanidou M. , Athanaselis S. , Spiliopoulou H.
Published: March 2009

Butyrylcholinesterase (BuChE) is routinely measured to assess exposure to or effects of organophosphorus insecticides (OP). As a biomarker, it can be used to clarify the relation between exposure to OP and health impairment. The interpretation of BuChE inhibition data, particularly of small changes in enzymatic activity, sometimes presents significant complexities. These complexities are presented in this short communication and the factors that influence the degree of BuChE inhibition are discussed. Despite the complexities of their interpretation, BuChE measurements remain a mainstay for the fast initial screening of exposure to OP; thus, they are a useful tool in the protection of humans, domestic animals and wildlife from overexposure to these toxic agents.
A prospective, non‐randomized trial comparing robot‐assisted laparoscopic and retropubic radical prostatectomy in one European institution
Authors: Ficarra Vincenzo , Novara Giacomo , Fracalanza Simonetta , D’Elia Carolina , Secco Silvia , Iafrate Massimo , Cavalleri Stefano , Artibani Walter
Published: March 2009

OBJECTIVE

To compare the functional results of two contemporary series of patients with clinically localized prostate cancer treated by robot‐assisted laparoscopic prostatectomy (RALP) or retropubic radical prostatectomy (RRP).

PATIENTS AND METHODS

This was a non‐randomized prospective comparative study of all patients undergoing RALP or RRP for clinically localized prostate cancer at our institution from February 2006 to April 2007.

RESULTS

We enrolled 105 patients in the RRP and 103 in the RALP group; the two groups were comparable for all clinical and pathological variables, except median age. For RRP and RALP the respective median operative duration was 135 and 185 min (P < 0.001), the intraoperative blood loss 500 and 300 mL (P < 0.001) and postoperative transfusion rates 14% and 1.9% (P < 0.01). There were complications in 9.7% and 10.4% of the patients (P = 0.854) after RRP and RALP, respectively; the positive surgical margin rates in pT2 cancers were 12.2% and 11.7% (P = 0.70). For urinary continence, 41% of patients having RRP and 68.9% of those having RALP were continent at catheter removal (P < 0.001). The 12‐month continence rates were 88% after RRP and 97% after RALP (P = 0.01), with the mean time to continence being 75 and 25 days (P < 0.001), respectively. At the 12‐month follow‐up, 20 of 41 patients having bilateral nerve‐sparing RRP (49%) and 52 of 64 having bilateral nerve‐sparing RALP (81%) (P < 0.001) had recovery of erectile function.

CONCLUSIONS

RALP offers better results than RRP in terms of urinary continence and erectile function recovery, with similar positive surgical margin rates.