bluuebunny/crossref_metadata_2025 huggingface dataset:
Bone Marrow Fractionation by the Haemonetics System: Reduction of Red Cell Mass before Marrow Freezing, with Special Reference to Pediatric Marrow Volumes
Authors: Saarinen U.M. , Lähteenoja K.M. , Juvonen E.
Published: March 2009

For purposes of freezing autologous marrow or transplants of allogeneic marrow with major ABO blood group incompatibility, 54 freshly harvested bone marrows from children of 7–65 kg of weight were depleted of their red cells with the Haemonetics V50 system. The marrow volumes ranged from 230 to 1,145 ml, with 17 small (200–399 ml), 18 intermediate (400–799 ml) and 19 large (800–1,200 ml) volumes. After processing, the median recoveries were: volume 24%, red cell mass 18%, and nucleated cells 75%. In the small marrow volume group, a good nucleated cell recovery was achieved at the expense of red cell depletion. The colony‐forming units, granulocytes‐macrophages (CFU‐GM) were normal after thawing of processed, cryopreserved marrows, and good engraftment of both allogeneic and autologous marrows were achieved. We conclude that marrow processing with the Haemonetics V50 system results in adequate red cell depletion and good nucleated cell recovery without open‐air contact of marrow or excessive time consumption. For small marrow volumes, however, the red cell depletion was suboptimal, and a bowl size smaller than 125 ml is desirable for pediatric use.
Transfusion Results of Filtered and Subsequently Stored Random Platelet Suspensions Prepared from Buffy Coats
Authors: Novotny V.M.J. , van Doom R. , Rozier Y. , D'Amaro J. , Harvey M.S. , Brand A.
Published: March 2009

There is almost general agreement that removal of leukocytes from blood components reduces the incidence of HLA‐antibody formation and refractoriness to random platelet transfusions. Recently filters have become available, which are able to reduce leukocyte contamination in platelet suspensions with acceptable platelet loss. We evaluated a cellulose acetate (CA) and a polyester (PE) filter, and stored buffy coat‐derived platelet suspensions after filtration. Both filters are effective for the removal of leukocytes to levels below 5 times 106 per transfusate. For the CA filter, platelet recovery was 73+13% yielding 256+53 times 109 platelets per transfusate from 6 donors. For the PE filter, platelet recovery was 90+9% and 327+51 times 109 platelets per transfusate. When a loading dose of less than 5 times 108 leukocytes was applied, 98% of the CA‐filtered suspensions and 100% of the PE‐filtered suspensions contained less than 5 times 106 residual leucocytes. In 123 patients transfusion results of CA‐filtered platelet suspensions stored for 72 h, were compared with those obtained by non‐stored, non filtered, random platelet suspensions which had been leukocyte depleted by differential centrifugation. Platelet increments 1 and 20 h after transfusion showed no statistical difference between CA‐filtered platelet transfusions stored for 72 h and non‐stored, non‐filtered platelet transfusions. In a new cohort of 117 patients, two filters and various postfiltration storage times were compared. Using both filters, the 1‐hour posttransfusion increments decreased to approximately 60% after 96 h of storage compared to results of storage periods of 72 h or less. Moreover, gradually decreasing platelet survical, as measured from 20‐hour post‐transfusion increments, was observed with increased storage time, from 24 h onwards. This decrease in platelet survival was associated with a decrease in pH. Because the pH and storage interval are strongly related, pH could not be evaluated as an independent parameter. The storage‐related decreased platelet survival, however, was more severe after PE filtration than after CA filtration, whereas pH was better maintained after PE filtration. In 164 cases the bleeding time was measured after transfusion of platelet stored for ≥72 h. Provided that a posttransfusion increment of ≥15 times 109/l was reached after transfusion with filtered, stored platelets, bleeding times shortened, indicating the hemostatic activity of filtered and subsequently stored platelets.
Granulocyte Serology Findings in Juvenile Symptomatic Idiopathic Autoimmune Neutropenia Using a Multiassay Procedure: Report on 21 Cases
Authors: Borzini Piero , Assali Giorgio
Published: March 2009

The serological findings on 21 children with idiopathic autoimmune neutropenia are reported. A multiassay procedure was adopted including agglutination, immunofluorescence and cytotoxicity tests. Beside a cause‐effect correlation between granulocyte antibodies and clinical course, a serologic polymorphism was found. As the sensitivity of each assay seemed to be related to the characteristics of the involved antigens and antibodies, as well as to the intrinsic sensitivity of the tests, the performance of a multiassay procedure appears to be advisable for the diagnosis and the follow‐up of autoimmune neutropenia.
Immunoglobulin Classes and Subclasses of Platelet Antibodies in a Case of Post‐Transfusion Purpura
Authors: Porretti L. , Marangoni F. , Cofrancesco E. , Neri M.C. , Rebulla P.
Published: March 2009

A multiparous white woman developed severe thrombocytopenia following transfusion of red blood cells. Six ineffective platelet transfusions (a total of 42 random donor concentrates) were given from day 0 to day +6, high‐dose steroids from day +1, progressively tapered until day +30, and a total of 150 g of intravenous immunoglobulins from day +2 to day +6. As platelet count had not increased significantly by day +8, four plasma exchange procedures, each consisting of 2,000 ml of plasma exchanged with 5% albumin solution, were performed on days +8, +10, +14 and +18. Platelet count was 5, 50, 100 and 234 times 109/1 on days +8, +14, +26 and +32 (discharge), respectively. The patient's acute phase serum contained increased levels of platelet alloantibodies with anti‐HPA‐la (PlA1, Zwa) specificity and a titer of 3,200. IgG1, IgG2 and IgG3 subclasses of platelet‐reactive antibodies in the patient's serum were elevated, whereas IgG4, IgM and IgA were within the reference values. Levels of IgG1, IgG2 and IgG3 of antiplatelet antibodies showed a marked and parallel reduction during treatment, but were still above the reference values at the end of treatment and 1 year later, when the patient platelet count was normal. Although a failure of intravenous immunoglobulins cannot be proven in this case, plasma exchange seems to have contributed more than intravenous immunoglobulins to clinical remission.
Do HLA Antigens Play a Role in Intermediate Uveitis?
Authors: Arocker‐Mettinger E. , Mayr W.R. , Huber‐Spitzy V. , Steurer‐Georgiew L. , Grabner G.
Published: March 2009

A total of 52 nonrelated Caucasian patients (21 male, 31 female, age at onset of the disease: 7–74 years, average: 31.6 years) suffering from intermediate uveitis were tissue typed for 56 different HLA‐A, ‐B, ‐C and ‐DR‐antigens.

No significant association between any specific HLA‐antigen and the disease, both when comparing the total patient group with a large, healthy control population, or any of the different subgroups (male vs. female, early vs. late onset, mild vs. severe course) could be demonstrated.
Correlation between Anti‐HBc Titers and HBV DNA in Blood Units without Detectable HBsAg
Authors: Iizuka Hisao , Ohmura Kazuyo , Ishijima Ayako , Satoh Koei , Tanaka Takeshi , Tsuda Fumio , Okamoto Hiroaki , Miyakawa Yuzo , Mayumi Makoto
Published: March 2009

Hepatitis B virus (HBV) DNA was tested for in 294 blood units which had antibody against hepatitis B core antigen (anti‐HBc) as the isolated serological marker of HBV infection. After amplification by polymerase chain reaction, HBV DNA was detected in 12 (6.9%) of 175 units that were positive for anti‐HBc with hemagglutination inhibition titers ≧26, significantly more often than in none of 119 units with titers ≦25 (p<0.01). These results indicate that the exclusion of blood units with isolated high‐titer anti‐HBc would be effective for further decreasing the risk of post‐transfusion hepatitis B.
Thrombocytopenia Due to Platelet Glycoprotein IIb/IIIa‐Reactive Autoantibodies Non‐Reactive with Platelets from EDTA Blood
Authors: Kuijpers R. W. A. M. , Ouwehand W. H. , Peelen W. , Michiels J. J. , Engelfriet C. P. , von dem Borne A. E. G. Kr.
Published: March 2009

We describe 2 patients with thrombocytopenia and autoantibodies in the blood against platelet membrane glycoprotein (GP) IIb/IIIa, in whom reactivity of the antibodies with their antigen was greatly diminished in the presence of sodium EDTA. One patient also showed a marked thrombocytopathy, suggesting a functional blocking of GP IIb/IIIa by these autoantibodies. These observations show that some immunologically important autoantibodies can easily be missed in routine serological investigations in which EDTA blood is used, obscuring the autoimmune nature of a thrombocytopenia and possibly of a thrombopathy as well.
Plasma Levels of the Lipid Mediators, Leukotriene B4 and Lyso Platelet‐Activating Factor, in Intraoperative Salvaged Blood
Authors: Sieunarine K. , Lawrence‐Brown M. M. , Goodman M. A. , Prendergast F. J. , Rocchetta Silvana
Published: March 2009

It has been shown that white cells and platelets release their granules into the plasma of salvaged blood. Whether this release is due to destruction alone or a combination of destruction and activation is not known. Lipid mediators, platelet‐activating factor and leukotriene B4, are produced by activated white cells and platelets and have effects on the cardiovascular, respiratory and immune systems and the microcirculation. The aim was to determine if white cells and platelets are activated in salvaged blood by measuring the levels of these lipid mediators. Ten patients undergoing aortic surgery, where intraoperative salvage was used, were studied. Blood samples were taken from the patient's circulation and the salvaged blood before and after washing. The levels of leukotriene (LTB4) and lyso platelet‐activating factor (PAF, the stable degradation product of PAF) were measured in the samples by a radioimmunoassay and a bioassay, respectively. The levels of both these substances increased in the unwashed salvaged blood (mean patient levels: LTB4 27±4.3 ng/ml and L‐PAF 73±8.5 ng/ml; mean unwashed blood levels: LTB4 95±12.2 ng/ml and L‐PAF 172.9±26.4 ng/ml) and were reduced by washing of the collected blood (mean washed blood levels of LTB4, 23.9±4.8 ng/ml, and L‐PAF 18±5 ng/ml).

The increase of the lipid mediators in the unwashed salvaged blood indicates that white cells and platelets are activated and releasing lipid mediators. Washing of the collected blood is effective in removing the lipid mediators.
Risk Factors for Hepatitis C Virus Antibody Positivity in Blood Donors in a Low‐Risk Country
Authors: Kolho E. K. , Krusius T.
Published: March 2009

Demographic variables, sexual risk behavior and prevalence of parenteral risk factors were studied in 305 randomly selected donors seronegative for hepatitis C virus, in 170 randomly selected donors reactive on solely enzyme‐linked immunosorbent assay (ELISA C‐100), in 71 consecutive donors reacting indeterminately according to the second‐generation recombinant immunoblot assay (RIBA II) and in 46 consecutive donors found to be positive using the RIBA II. Donors who were positive by RIBA II had significantly more often a risk factor, for example use of intravenous drugs or previous blood transfusion, than donors reacting indeterminately (34 out of 46) (73.9%) versus 14 out of 71 (19.7%, p = 0.0000). Donors reacting indeterminately by RIBA II had one of those risk factors significantly more often than seronegative donors (14 out of 71) (19.7%) versus 23 out of 280 (7.8%, p<0.005). When donors either positive or indeterminate by RIBA II were compared with donors negative for hepatitis C antibodies, the odds ratio for a possible parenteral source of infection was 7.6 (p = 0.0000). Subjects who had received a poor education (odds ratio 0.3, p<0.001) or who lived in southern Finland (odds ratio 2.3, p<0.05) were also at higher risk for being positive or indeterminate in RIBA II. First‐time donors were also prone to having antibodies according to RIBA II (odds ratio 2.2, p = 0.1), whereas sexual risk behavior, gender, age, occupational class and type of residential area were not risk factors for hepatitis C antibodies in RIBA. It is concluded, that a possible parenteral source of infection is the major risk factor for having antibodies to hepatitis C virus as tested by RIBA.
Process‐Scale Purification of Immunoglobulin M Concentrate
Authors: Ng Paul K. , O'Rourke Peter E. , Andersen John D. , Tsay Grace C. , Dobkin Milton B.
Published: March 2009

An IgM concentrate was purified from Cohn fraction III. Efficiency of euglobin precipitation was shown to be controlled by pH and ionic strength. Prekallikrein activator activity in the product was insignificant. Overall yield from the octanoic acid supernate and purity of the concentrate were 66 ± 8 (n = 16) and 50 ± 5% (n = 16), respectively. Solvent‐detergent treatment to inactivate lipid‐enveloped viruses was demonstrated and implemented into the process. Process studies to control residual virucidal agents and C4a generating activity are presented.
Studies in Red Blood Cell Preservation
Authors: Greenwalt T.J. , Dumaswala U.J. , Dhingra N. , Allen C.M. , Silberstein E.B.
Published: March 2009

Studies were conducted to examine whether an experimental additive solution (EAS‐2) containing, in mM: 20 NH4Cl, 30 Na2HPO4, 2 adenine, 110 dextrose, 55 mannitol, pH 7.15, would be useful to extend the storage shelf life of human RBCs. With 6 pairs of split units, ATP concentrations were better maintained for 12 weeks with EAS‐2 than with Adsol® (1.8 vs. 1.1 μmol/gHb, respectively, p = 0.002). Autologous 24‐hours 51Cr viability values for split units in the same donors were: on 6 paired units at 8 weeks, EAS‐2 87.0±4.5%, Adsol 72.6±2.3%, p = 0.004; on 11 paired units at 9 weeks, EAS‐2 79.5±7.1%, Adsol 68.2±10.1%, p = 0.0003. The data suggest that packed RBCs stored for 9 weeks with EAS‐2 will be suitable for transfusion following the removal of supernatant with a single washing step.
Improved Removal of Anti‐A and Anti‐B Antibodies from Plasma Using Blood‐Group‐Active Haptens
Authors: Blomberg L. , Bratt T. , Chester M. A. , Hansson B.
Published: March 2009

Highly efficient anti‐blood group A and B immunoadsorbents for extracorporeal treatment were developed by immobilizing A and B trisaccharide and A tetrasaccharide haptens on Sepharose 4FF and Fractogel TSK using three different methods. The adsorption of the IgG and IgM anti‐A antibodies was essentially the same regardless of the A hapten used or the method of oligosaccharide coupling. The adsorption of IgM anti‐A was found to be more sensitive than IgG anti‐A to changes in column flow rate. The binding of both the IgM and IgG antibodies was slightly lower at 37°C than at 22°C. An anti‐A/anti‐B adsorbent column potentially suitable for treatment of patients was prepared. A column switching system resulted in a more efficient adsorption of antibodies. Hapten leakage from the column was very low. No nonspecific adsorption of plasma proteins to the column (other than traces of albumin) could be detected.
Direct Agglutination of Weak D Red Cells by Tetramolecular Complexes Containing Monoclonal IgG Anti‐D
Authors: Lemieux Réal , Verrette Sylvain , Broly Hervé , Perron Serge
Published: March 2009

The capacity of blood group antibodies to agglutinate red cells suspended in saline is largely dependent on the antibody isotype. The immunological cross‐linking of IgG antibodies has previously been described as a means to increase the reactivity of IgG in many situations. We have prepared anti‐D‐containing complexes by blending a human IgG anti‐D monoclonal antibody (mAb) and a murine anti‐human IgG mAb. In standard red cell serology assays, the anti‐D complexes exhibited a very high avidity and could agglutinate weak D‐positive red cells in direct saline testing. These results indicate that potent saline hemagglutinating reagents of RhD and eventually of other blood group specificities can be prepared from IgG mAbs.
Predictive Value of Screening Tests for Persistent Hepatitis C Virus Infection Evidenced by Viraemia: Japanese Experience
Authors: Watanabe J. , Matsumoto C. , Fujimura K. , Shimada T. , Nishioka K. , Yoshizawa H. , Okamoto H. , Iizuka H. , Tango T. , Ikeda H. , Endo N. , Mazda T. , Nojiri T. , Aoyama K. , Kanemitsu K. , Yamano H. , Mizui M. , Yokoishi F , Tokunaga K.
Published: March 2009

In November 1989, Japanese Red Cross Blood Centres started screening for heaptitis C virus (HCV) with enzyme‐linked immunosorbent assay (Elisa) for the C100‐3 viral peptide as the first such nationwide programme in the world. Thereafter post‐transfusion non‐A non‐B hepatitis (PTNANBH) was reduced by 61–80%, but this was not as complete a success as our programme to prevent post‐transfusion hepatitis B by screening for high titer hepatitis B core antibody, which we began in the same period. In order to acquire more effective control of PTNANBH, the HCV core‐related antigen (GOR, N14) and second‐generation Elisa (Ortho2, Abbott2)and second‐generation antigen agglutination (PA, PHA) tests have been employed. Among 16,500 donors in 11 blood centers, 365 were serologically positive by at least one of these tests. Among these, HCV RNA was detected in 138 units and the remaining 227 were HCV RNA negatives. The effectiveness of these serological tests to detect HCV RNA‐positive status were analyzed. Passive haemagglutination and particle agglutination (PHA and PA) tests were highly effective to predict HCV viraemia among blood donors. Also, these tests can easily determine antibody titre. By either PHA or PA, all units with ≧212 agglutination titre (120 and 122 units) were HCV RNA positive and all agglutination‐positive units with serum alanine aminotransferase level higher than 35 Karmen units were HCV RNA positive. These results have formed the basis for implementing a more effective screening for HCV viraemia in blood donors, where effectiveness is defined as enhancing the protection of patients from post‐transfusion hepatitis C and providing higher quality information to achieve more effective donor counselling.
Recombinant Human Erythropoietin as Adjuvant Treatment for Autologous Blood Donation: A Prospective Study
Authors: Beris P. , Soulier‐Lauper M. , Tullen E. , Hugli A. , Miescher P. A. , Mermillod B. , Levy G. , Laubriat M.
Published: March 2009

In a prospective randomized study we investigated the potential of subcutaneous recombinant human erythropoietin (rhEpo) as adjuvant treatment for autologous blood transfusions (3 units) in elective surgery. Four and 2 weeks before surgery, 49 patients received 6 times 10,000 U of rhEpo. ΔHb values (days ‐28 and 0) of the rhEpo group were compared to ΔHb values of 52 controls (no rhEpo). Reticulocytes were measured at days ‐21, ‐14, ‐7 and 0. Peri‐ and postoperative supplementary homologous blood requirements were compared in the two randomized groups. ΔHb of rhEpo group was 0.96 g/dl (mean value) and 2.38 for controls. Reticulocyte count increased earlier and to higher levels in rhEpo‐treated patients. Except in 1 case, Epo was well tolerated. These results indicate that autologous predonation (3 times 400 ml) does not create anemia if adjuvant Epo treatment is given. However, homologous blood requirements were not significantly different, which is probably due to the fact that 96 of the 101 treated patients underwent elective orthopedic surgery requiring limited blood replacement. Significant benefit of the Epo regimen can be expected in elective cardiovascular and hepatic surgery where larger amounts of blood (5–6 units) are needed.
Effect of Prestorage Leukocyte Reduction on Proteins of Platelets Obtained by Apheresis
Authors: Sarraj‐Reguieg Amira , Tissot Jean‐Daniel , Hochstrasser Denis F. , von Fliedner Vladimir , Bachmann Fedor , Schneider Philippe
Published: March 2009

The effect of prestorage leukocyte reduction was evaluated on platelet concentrates (PCs) obtained by apheresis using the ‘surge’ technique. Two hours after collection, the PCs were divided into 2 equal units. One unit was filtered through a Sepacell PL‐10a®, producing a filtered PC (FPC). The second unit constituted a non‐filtered PC (NFPC). FPCs and NFPCs were stored at room temperature in 1,400‐ml CLX® bags on a horizontal agitator up to 7 days. We analyzed platelet samples obtained during storage from NFPCs and FPCs at days 1,3 and 7. The expression of membrane glycoproteins (GP)Ib and GPIIb/IIIa (assessed by flow cytometry), platelet response to thrombin and ristocetin (aggregometry) and global platelet protein pattern (studied by high‐resolution two‐dimensional gel electrophoresis) remained stable over the 7 days of storage in NFPCs as well as in FPCs. However, in both preparations, the expression of GMP‐140 (flow cytometry) progressively increased during storage. Our in vitro study indicates that early leukocyte reduction by filtration of apheresis PCs does not induce modifications in platelet GPs and protein patterns.
Systematic Use of Leukocyte‐Free Blood Components to Prevent Alloimmunization and Platelet Refractoriness in Multitransfused Children with Cancer
Authors: Saarinen U. M. , Koskimies S. , Myllylä G.
Published: March 2009

A major drawback in the use of platelet transfusions is the development of platelet refractoriness that is usually caused by alloimmunization provoked by leukocytes present in the blood products. We evaluated the development of alloimmunization both by serological demonstration of HLA antibodies and by clinical demonstration of platelet refractoriness in 50 multitransfused children who exclusively received filtered, leukocyte‐free blood products. The basic diagnoses included acute leukemia, pediatric solid tumors, and severe aplastic anemia. We also present a reference group of 10 similarly multitransfused children, who either deliberately or inadvertently had received nonfiltered blood products on some occasions. In the study group of 50 children, none developed platelet refractoriness. The median corrected increment was 11.1 times 109/1 at the time of initial diagnosis, and 10.4 at the time of the study 8 months later (median). None had detectable HLA antibodies in serum. In contrast, in the reference group 3/10 had HLA antibodies in serum, and 1/10 was clearly refractory to random pooled platelets. We conclude that systematic use of leukocyte‐free blood components effectively prevents alloimmunization in heavily transfused children with cancer. Leukocyte depletion to the level of < 1 times 106 of contaminating leukocytes per unit is sufficient and seems to prevent alloimmunization totally.
Working Standards for Hepatitis B Surface Antigen for Use in the UK Blood Transfusion Service: Results of a Collaborative Study
Authors: Ferguson Morag , Pipkin Pamela A. , Heath Alan B. , Minor Philip D.
Published: March 2009

Twenty‐six laboratories including 22 Blood Transfusion Centres in England, Scotland, Wales and Northern Ireland participated in a collaborative study to assess the suitability of proposed working standards for hepatitis B surface antigens (HBsAg) for use in the UK Blood Transfusion Service. Data from 88 assays indicated that all laboratories could detect HBsAg in the proposed working standard at a concentration of 0.5 IU/ml. Samples containing HBsAg at a concentration of 0.125 IU/ml were scored positive in 80% of assays and it was concluded that this would be a suitable concentration for inclusion in a monitor sample which would indicate whether the sensitivity of an assay system in routine use is varying over a period of time. The use of a batch of ‘provisional’ working standard tested under field conditions over a period of 4 months has been monitored. HBsAg was not detected in only 2 out of 6,523 assays.
Inactivation of Viruses during Ultraviolet Light Treatment of Human Intravenous Immunoglobulin and Albumin
Authors: Hart H. , Reid K. , Hart W.
Published: March 2009

A comparison of ultraviolet (UV) irradiation at two wavelength ranges UVB (280–320 nm) and UVC (lower than 280 nm) showed that UVC in particular could very effectively inactivate, in intravenous immunoglobulin (IVIG) and albumin preparations, non‐enveloped and non‐acid labile model viruses (i.e. Polio 2 and T4 phage) and dry heat‐resistant viruses (vaccinia and T4 phage). This effective virucidal treatment (5 min, 5,000 J/m2 dose) was achieved before an unaceeptable level of WIG aggregates occurred. The use of UV irradiation to inactivate infectious agents could add safety and supplement current methods, e.g. solvent/detergent, low pH, which do not inactivate non‐enveloped, non‐acid labile or dry‐heat‐resistant viruses at present.
Second‐Generation Hepatitis C Elisa Antibody Tests Confirmed by the Four‐Antigen Recombinant Immunoblot Assay Correlate Well with Hepatitis C Viremia and Chronic Liver Disease in Swedish Blood Donors
Authors: Shev S. , Foberg U. , Fryden A. , Hermodsson S. , Lindh G. , Lindholm A. , Månsson A.S. , von Sydow M. , Weiland O. , Widell A. , Norkrans G.
Published: March 2009

Seventy‐three Swedish blood donors (52 men, 21 women; median age 36 years) repeatedly reactive for hepatitis C antibodies (anti‐HCV C‐100‐3) were tested with a second‐generation (2nd‐gen) anti‐HCV Elisa and a 4‐band recombinant immunoblot assay (RIBA 2). These results were correlated to serum alanine aminotransferase (S‐ALAT), liver morphology and viremia as detected by ‘nested’ polymerase chain reaction (PCR) based on primers from a 5′‐noncoding sequence of the HCV genome. Thirty‐five of 46 (76%) donors with positive 2nd‐gen Elisa tests confirmed by RIBA 2 were PCR positive whereof 27 had histological findings compatible with chronic persistent hepatitis (CPH) and 7 had chronic active hepatitis (CAH). Ten of 56 (18%) 2nd‐gen Elisa‐positive donors were RIBA 2 negative (or indeterminate) and none of these had chronic hepatitis nor were PCR positive. Seventeen of 73 (23%) donors were 1st‐gen Elisa positive but 2nd‐gen Elisa negative. All of these were PCR negative and only 1 (6%) had chronic hepatitis (CPH). An elevated S‐ALAT level (reference <0.7 μkat/1) was found in 26 2nd‐gen Elisa and RIBA 2‐positive donors of which 18 had CPH and 7 had CAH and all 25 were PCR positive. A normal S‐ALAT level was found in 9 of 34 (26%) donors with chronic hepatitis (all had CPH) and positive PCR. We have found that blood donors with positive 2nd‐gen anti‐HCV Elisa tests confirmed by RIBA‐2 and especially with a concomitant elevated S‐ALAT are highly likely to be viremic as demonstrated by PCR and to have chronic hepatitis.
Transfusion‐Transmitted Yersinia enterocolitica Infection: Protection Through Buffy Coat Removal and Failure of the Bacteria to Grow in Platelet‐Rich or Platelet‐Poor Plasma
Authors: Gong J. , Högman C.F. , Hambraeus A. , Johansson C.S. , Eriksson L.
Published: March 2009

In a previous study, removal of white blood cells (WBC), by filtration 5 h after deliberate contamination of whole blood with a type 0:3 strain of Yersinia enterocolitica, was shown to be an effective way of avoiding bacterial growth in red blood cells (RBC) during storage. In the present study the Opti‐System technique was used to remove the buffy coat from whole blood, leaving 10–20% of the original number of WBC in the RBC preparation. In one series of experiments, all of 4 units of RBC suspension, from which buffy coats were removed 2 h after inoculation of 112 colony‐forming units (cfu) per ml of Y. enterocolitica, became Yersinia‐free, while abundant bacterial growth occurred in all of 4 units where RBC suspension and buffy coat had been recombined. In a second series of 10 experiments, with an inoculum of 80 cfu/ml, no growth was found in platelet‐poor plasma stored for 42 days at 4°C. Five out of 10 RBC suspensions in SAGM additive solution remained Yersinia‐free throughout a 6‐week storage period; 4 of these 10 units showing growth of Yersinia after 4 weeks and 5 after 6 weeks. In the buffy coats bacterial growth was found in 1 out of 10 units after 1 week, 4 after 2 weeks, and in all of 10 units after 4 weeks. In 2 control experiments with WBC‐reduced RBC inoculated with the same bacterial dose, growth started within 24 h and was abundant after 1 week.

In platelet‐rich plasma (PRP), prepared from 4 units of whole blood that had been inoculated with Y. enterocolitica (106 cfu/ml), no bacterial growth was found during 10 days of storage at 22°C. Also, after Y. enterocolitica inoculation (114 cfu/ml) directly into PRP, bacteria failed to grow.

We conclude that removal of the buffy coat from whole blood that had been previously contaminated with a moderate inoculation of Y. enterocolitica partly protects the red cells from bacterial growth during storage. The risk of transmitting Y. enterocolitica with platelet‐poor plasma, liquid stored at 4°C, seems minimal. This bacterial species shows very poor capacity to grow in PRP at 22°C.
Protection Against Rh D‐Haemolytic Disease of the Newborn by a Diminished Transport of Maternal IgG to the Fetus
Authors: Dooren Marion C. , Engelfriet C. Paul
Published: March 2009

IgG present in the circulation of newborn infants is predominantly of maternal origin. In cases of severe Rh D alloimmunization, the maternal anti‐D antibodies may lead to Rh D‐haemolytic disease of the newborn. However, an impaired placental transport of maternal IgG could be associated with the protection of the fetus/newborn against haemolytic disease as shown in this case report.
Antibodies to Platelet Glycoprotein V in Polytransfused Patients with Haematological Disease
Authors: Meenaghan M. , Judson P. A. , Yousaf K. , Lewis L. , Pamphilon D. H.
Published: March 2009

Polytransfused patients often develop platelet‐reactive antibodies (PRAb). These give positive reactions in the platelet immunofluorescence test (PIFT) and may be either lymphocytotoxic (LCTAb) or platelet‐specific antibodies (PSAb). The latter may be detected in the PIFT using chloroquine‐treated platelets (Chl‐PIFT) or by immunoblotting. Serial samples from 106 multiply transfused patients with bone marrow failure were screened by PIFT using a microplate method and flow‐cytometric analysis. PSAb activity was confirmed by Chl‐PIFT. In 45 (42%) of the patients studied PSAb were detected; 37 (35%) formed LCTAb and 19 (51%) had co‐existent PSAb. Sera from 25 of 27 patients with a positive Chl‐PIFT, retested by immunoblotting, recognised determinants of Mr 82–160 kD on whole platelets. A large group became sensitised to a component of Mr 105–115 kD reduced (99 kD non‐reduced) with similar electrophoretic mobility to GPIIIa using a monoclonal anti‐GPIIIa and two human polyclonal anti‐HPA‐1a sera; some also produced anti‐GPIIb. The largest group recognised a determinant of Mr 80–83 kD, probably glycoprotein V (GPV). Three sera were immunoblotted against thrombin‐treated platelets and the results confirmed GPV specificity.
Platelets Stored for 6 Days in a Polyolefin Container in a Synthetic Medium with Minimal Plasma Carry‐Over
Authors: Boomgaard M. N. , Gouwerok C. W. N. , De Korte D. , Loos J. A.
Published: March 2009

Platelet concentrates (PC) were stored for 6 days in either polyolefin (PO) or polyvinylchloride/di‐(2‐ethylhexyl)phtalate (PVC/DEHP) bags in 100% plasma or in a synthetic medium with 35 or 10% plasma. For all conditions studied the usual in vitro parameters were well maintained, with a pH above 6.8. In both bag types platelets can be satisfactorily stored for 6 days in a synthetic medium with minimal amounts of residual plasma. For this medium, the PO bag offers a slight advantage with respect to the preservation of platelet ATP content (>80 versus >70% in the PVC bags) and aggregation and adhesion capacity. The adhesion capacity increased in the PO bags, while it decreased in the PVC bags.
A Novel Variant of the Human Blood Group K1 Antigen
Authors: Skradski K. , Reid M. E. , Mount M. , Polesky H.F. , Sausais L. , Yacob M. , Batts R.
Published: March 2009

A discrepancy in duplicate anti‐K1 typing in a parentage case led to the discovery of an unusual K1 blood group antigen. Red blood cells from the propositus (JC) express a rare variant of the K1 antigen that is detectable by only 8 of 72 sera containing anti‐K1. Absorption and elution studies using reactive anti‐K1 confirmed the presence of a K1 antigen. Nonreactive anti‐K1 was not absorbed by or eluted from JC's red blood cells. Red cells from 3 of the propositus's siblings also had the variant K1 antigen. The variant antigen exhibited qualitative as well as quantitative differences as compared to normal K1, and we have named it K1var.
Postnatal Development of Red Cell Leaand LebAntigens in Chinese Infants
Authors: Lin Marie , Shieh Shiow‐Hua
Published: March 2009

Lewis phenotyping of 487 blood samples from Chinese newborn infants and young children, revealed that 50% of cord cells were Le(a–b+) and 50% Le(a–b–). The weak Lebantigen of Le(a–b+) cord cells is most likely produced by the newborn infant rather than of maternal origin and it appears that these infants eventually develop by way of an intermediate Le(a+b+) stage into the adult Le(a–b+) phenotype. Most infants with Le(a–b–) cord cells, but not all, appear to develop through a transitional Le(a+b–) stage, into Le (a+b+) by about 1 month of age, most likely continuing as such into adulthood. This development of Le(a–b–) cord cells into the adult Le(a+b+) phenotype is postulated to be the result of the weak secretor geneSeω. Those infants with Le(a–b–) cord cells that do not convert to Le(a+b+) during the first month of life, most likely remain as such into adulthood. The blood of 120 adult voluntary blood donors, used as controls, reconfirmed adult Chinese phenotypic frequencies of approximately 70% Le(a–b+), 22% Le(a+b+) and 8% Le(a–b–).
Relative Efficiency of Leucocyte Removal Procedures for the Production of Leucocyte‐Poor Red Cell Concentrates Assessed by Flow Cytometry
Authors: Farrugia A. , Tan Y. , Romeo A. , Martin L. , Rolland J. R. , Kellner S. , Piouronowski H.
Published: March 2009

Flow cytometry was used to: (1) determine residual leucocyte numbers in red cell suspensions following the range of leucocyte depletion procedures used in our organisation, and (2) to characterise phenotypically the leucocytes using direct immuofluorescence with monoclonal antibodies to cell surface receptors. Under the conditions used, a lower limit of detection of 2.5 leucocytes per μl (equivalent to 3.43 log10 or 99.96% removal) could be achieved. Filtration through polyester filters was found to remove up to >99.96% of the initial leucocytes; however, a significant differential efficacy was observed between filters from different manufacturers even when filters with similar costs were compared. The order of filter brands with respect to leucocyte removal found was Pall BPF4 = Erypur Optima G‐O > Sepacell R500 > Pall RC50. Pheno‐typing revealed that increasing filtration efficacy was associated with a preferential removal of lymphocytes; conversely, a second filtration over one brand of filter allowed proportionately more lymphocytes to pass through compared with the first filtration. A saline wash following filtration removed a further 0.5% of the initial leucocyte content, and was associated with a preferential loss of granulocytes. Freeze‐thawing the red cell suspension removed fewer leucocytes (96.3%) than did filtration (98.74% to>l99.6%) or filtration followed by washing (99.22%), and also led to preferential loss of granulocytes. Flow cytometry provides a reliable tool for the quality control of leuco‐depleted red cells, and allows a qualitative assessment of the residual leucocytes. This information is of value in choosing procedures aimed at decreasing the risk of alloimmunisation and post‐transfusion reactions.
Patterns of Autologous Blood Use in Elective Orthopedic Surgery: Does the Availability of Autologous Blood Change Transfusion Behavior?
Authors: Julius Carmen J. , Purchase Kimberly S. , Isham Betsy E. , Howard Phillip L.
Published: March 2009

We studied the orthopedic surgery service at our institution to determine whether the mere availability of autologous blood (AB) affected transfusion practice. As a group, patients who had AB available received an average of 1.11 fewer red cell units per hospitalization than did patients with only homologous blood (HB) available. At every transfusion episode, those patients having AB available received fewer red cell units than did patients without AB available. Predeposit of autologous red cells was effective in protecting 77.6% of patients from HB exposure. The availability of autologous red cells resulted in an overall more conservative approach to transfusion.
Blood Product Utilization in Hip and Knee Arthroplasty: Effect of Gender and Autologous Blood on Transfusion Practice
Authors: Churchill W. H. , Chapman Richard H. , Rutherford Cynthia J. , Poss Robert , Wallace Edward L. , Surgenor Douglas MacN.
Published: March 2009

Analysis of total blood product support for a 1‐year cohort of patients undergoing hip or knee total joint arthroplasty showed significant differences in transfusion therapy between patients who predeposited autologous blood and those who did not. In primary joint arthroplasty, 51% of nonpredepositing patients undergoing hip replacement and 28% of nonpredepositing patients undergoing knee replacement required red cell transfusions. In revision procedures, 58–61% were transfused. Predepositors requiring only autologous blood received less blood per patient than nonpredepositors; however, 73–87% of primary and 86–88% of revision arthroplasty patients were transfused. Predepositors receiving supplemental allogeneic blood used a volume of red cells comparable to nonpredepositing patients, which was significantly greater than the red cell requirement of predepositors using only autologous blood. Moreover, regardless of predeposit status, the extent of red cell replacement differed between men and women. Male patients presented with significantly higher hematocrits and were less likely to be transfused than females undergoing the same procedure. However, once the transfusion‐decision was made, the average amount of red cells given for each procedure did not show gender‐related variation. Despite differences in admission and lowest observed hematocrits, all patients were discharged with hematocrits in the same range, suggesting that men were replaced with relatively less blood than women. These differences in transfusion practice relating to gender and predeposit status could not be associated with identifiable changes in clinical outcome which might provide rationale for the observed differences in practice.
The First Reported Case of Anti‐Dob Causing an Acute Hemolytic Transfusion Reaction
Authors: Halverson G. , Shanahan E. , Santiago I. , Mabile R. , Thurrell T. , Strupp A. M. , Wolf C. F. W. , Spruell P. , Moulds M. K.
Published: March 2009

The antibodies of the Dombrock blood group system have only rarely been encountered in transfusion practice, and anti‐Dob has not previously been implicated in an acute hemolytic transfusion reaction. We have encountered the first such case involving a chronically transfused black female with hemoglobin SS disease and multiple antibodies in her serum. During a previous admission for sickle cell crisis, the patient received 3 units of compatible blood with no untoward effects. Serum obtained 21 days later contained, in addition to the known antibodies, anti‐S plus an unidentified antibody showing characteristics of HTLA. Blood lacking the E, K1, Fy(a), Jk(b) and S antigens was obtained, and 2 least incompatible units were transfused. While administering the second unit, the patient complained of fever and low back pain, and hemoglobinemia was detected. Anti‐Dob was identified in the post‐reaction samples by absorption‐elution tests, and the patient was confirmed to be Do(a+b–). The first unit transfused during this hemolytic episode tested Do (b+). This case, and a similar case involving anti‐Doa reported in 1986, strengthens the belief that Dombrock antibodies are clinically significant and illustrates the need for their differentiation, prior to transfusion from less clinically significant HTLA antibodies.
Rh18 and hrS Blood Groups and Antibodies
Authors: Moores Phyllis
Published: March 2009

Anti‐hrS, also known as the Shabalala antibody, is unlikely to be found in unabsorbed human serum. The term ‘anti‐hrS’ was devised by Shapiro in 1960 to describe the antibodies remaining in the absorbed serum after anti‐Rh18 had been absorbed with R2R2 red cells. R2R2‐absorbed anti‐Rh18 (anti‐hrS), although an interesting research tool, is therefore clinically irrelevant. Unabsorbed anti‐Rh18, on the other hand, is a clinically significant antibody. It is compatible not only with Rh‐‘deleted’ and Rhnull red cells, as described by Shapiro, but is also compatible with the red cells of numbers of Southern African Blacks and Coloureds (mixed race) who have R0, R0u or R2r pheno‐types. Anti‐Rh18 causes haemolytic disease of the newborn and, when uncontaminated with other antibodies, is a further reagent for resolving Rh grouping problems.
M9+ WINS Blood Group Phenotype: Further Observations
Authors: Green Carole , Daniels Geoff , Skov Flemining , Tippett Patricia
Published: March 2009

Red cells carrying the low‐frequency MNS antigen M8 reacted with the only example of anti‐DANE, an antibody which had previously defined the GP.Dane (Mi.IX) phenotype. Furthermore, Mg + cells reacted with the original anti‐Mur (serum of Mrs. Murrell), but with none of 14 other anti‐Mur. Therefore, M8 + cells carry both DANE antigen and an atypical Mur antigen. Immunoblotting of membranes from Mg + cells with anti‐M, and with eluates prepared from anti‐Mg and Mrs. Murrell's serum demonstrated a glycophorin A (GPA) molecule whose mobility was increased by an apparent Mr of about 3,000 presumably due to the loss of the three O‐glycans known to be absent from Mg ‐active GPA.
Studies of One Invasive and Two Noninvasive Methods for Detection of Bacterial Contamination of Platelet Concentrates
Authors: Högman Claes F. , Gong Jian
Published: March 2009

Recently, a new method for noninvasive detection of bacterial contamination of platelet concentrates was described [Arpi et al: Vox Sang 1993;65:335–336]. A CO2‐sensitive label is applied on the outer surface of the plastic container of the platelet concentrate. When any contaminating bacteria have increased the pCO2 to a certain level the label changes color, indicating bacterial growth. We have studied this technique and applied such sensitive labels both directly onto PCs and onto plastic bags with culture medium into which platelet contents have been transferred. However, in both cases the sensitivity of the technique was found insufficient for practical quality control of PCs. There seem to be two problems with this principle: (1) platelets produce CO2 and (2) a considerable amount of the gas passes beside the detection label, in this way decreasing the sensitivity of detection by the label. This noninvasive technique was compared with an invasive method using automated bacterial culture, which was found sensitive and rapid.
A Highly Purified Antithrombin III Concentrate Prepared From Human Plasma Fraction IV‐1 by Affinity Chromatography
Authors: Lebing Wytold R. , Hammond David J. , Wydick James E. , Baumbach George A.
Published: March 2009

We describe an improved method for large‐scale purification of antithrombin III (AT‐III) from human plasma involving heparin affinity chromatography of redis‐solved fraction IV‐1 paste, viral inactivation by heating, followed by a second heparin affinity column. The characteristics of a new heparin affinity resin and the ability to extrapolate process behavior from small‐scale (20 ml) to large‐scale (40 liter) columns are described. This supports the use of the small‐scale column for process optimization and validation studies in compliance with current regulatory requirements for biological products. The process has been characterized by analytical techniques including sodium dodecyl sulfate (SDS), reducing SDS, and nondenaturing polyacrylamide gel electrophoresis; laser desorption time‐of‐flight mass spectroscopy, and electrospray mass spectroscopy. These results demonstrate that greater than 95% of the protein in the final products is AT‐III, which is greater than 95% active as defined by thrombin inhibition.
Studies in Red Blood Cell Preservation. 8. Liquid Storage of Red Cells in a Glycerol‐Containing Additive Solution
Authors: Dumaswala U.J. , Bentley N.L. , Greenwalt T.J.
Published: March 2009

The purpose of the present study was to determine whether a hypotonic additive containing a low concentration of glycerol as a membrane permeable solute would improve the liquid storage of red blood cells (RBCs). Packed RBCs were stored either with 200 ml of an experimental additive solution, EAS 25, containing (mM): glycerol 150, adenine 2, glucose 110, mannitol 55, and NaCl 50, or with 100 ml/unit of a conventional additive solution Adsol®. The results show that the adenosine triphosphate values, hemolysis, potassium leakage, and the morphology scores of RBCs were significantly better with EAS 25 than with Adsol up to 84 days of storage. The ATP values were significantly different only after the first 42 days of storage. The mean corpuscular volumes (MCVs) of the RBCs were significantly higher throughout in the experimental additive accompanied by decreased microvesiculation as compared to Adsol. The total microvesicle membrane protein shed by 100 ml of RBCs was 47.92±12.31 mg in Adsol and 18.96±5.49 mg in EAS 25 (p<0.001). The larger MCVs of the RBCs in EAS 25 may have a favorable effect on maintaining membrane integrity by decreasing the loss of membrane by microvesiculation.
Complement Activation during Storage of Single‐Donor Platelet Concentrates
Authors: Schleuning Michael , Böck Markus , Mempel Wolfgang
Published: March 2009

Single‐donor platelets are stored up to 5 days prior to transfusion. Since contact of plasma to plastic surfaces may lead to complement activation, we investigated whether there is any increase in the complement factors C3a, C4a and C5a in routinely stored single‐donor platelet concentrates. C3a levels increased about 40‐fold during a 7‐day storage. C4a levels also increased with storage time but to a lesser extent. By contrast, C5a levels remained stable throughout this period. ADP‐ and collagen‐induced aggregation was impaired after storage of platelets, indicating severe functional injury. In platelet‐poor plasma stored under identical conditions a comparable increase in C3a and C4a concentrations was observed. The loss of platelet function during storage might at least in part be due to the excessive anaphylatoxin concentrations observed.
In vitro Platelet Function during Storage in Three Different Additive Solutions
Authors: Koerner K. , Sahlmen P. , Zimmermann B. , Cardoso M. , Kubanek B.
Published: March 2009

Three different synthetic media without glucose were studied for platelet storage. The first medium contained acetate and gluconate. The second contained acetate, gluconate and citrate. Finally the third contained phosphate and mannitol. The purpose of the study was to investigate whether there were differences among the various media in terms of preservation of platelet quality. Pools of platelet concentrates were prepared from buffy coats. In vitro function and metabolic parameters were measured during 5 days of storage in these additive solutions as well as in plasma. Platelet aggregation, hypotonic shock response and release of β‐thromboglobulin, platelet factor 4 and lactate dehydrogenase of the cytosol were equivalent in the media containing acetate compared to plasma storage. In vitro platelet functions and pH in these two media were better preserved compared to the medium with phosphate and mannitol. In addition bacteriological studies using platelets suspended in additive solutions or in plasma were carried out. Carryover of 20% of plasma to the synthetic media necessary for successful platelet storage in these additive solutions allows bacteriological growth. As shown, inoculation of 1 colony/ml Staphylococcus epidermidis leads to 106–107 organisms/ml after 5 days of storage.