bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro and in vivo Comparison of Platelet Concentrates Collected by Automated versus Manual Apheresis
Authors: Ross D. G. , Holme S. , Heaton W.A. L.
Published: March 2009

Abstract. Platelet concentrates collected by continuous flow automated apheresis (Fenwal CS‐3000) were compared with those collected by manual apheresis to determine whether the prolonged centrifugation and vigorous resuspension affected platelet viability and in vitro function. Paired autologous reinfusion studies (111Indium) of 5 normal donors showed no significant differences in the mean percent recoveries (50.8±7.0% vs 53.8 ±4.0%) or survivals (174±23h vs 188±10h) for platelets collected by manual versus automated apheresis. Platelets collected by automated apheresis had a significantly higher level of β‐thromboglobulin release, but there were no significant differences between platelets collected by the two methods in regard to other in vitro parameters (ATP levels, LDH released, hypotonic shock response) believed to reflect platelet activation, injury, and malfunction. These results suggest that only slight activation of platelets takes place during automated apheresis.
Development of a Direct‐Binding Enzyme‐Linked Immunosorbent Assay as an Alternate Assay for Anti‐Hepatitis B Core
Authors: Yang Neng Yu , Bryant Jeffrey L. , Shih J. Wai‐Kuo
Published: March 2009

Abstract. Anti‐hepatitis B core (anti‐HBc) screening is now used in some countries as a surrogate test to reduce the incidence of posttransfusion non‐A, non‐B hepatitis. The purpose of this study was to develop and standardize an alternate assay, for the detection of anti‐HBc, based on a direct‐binding enzyme‐linked immunosorbent assay (db‐ELISA). Microtiter plates were coated with recombinant DNA hepatitis B core antigen. Patient antibody was detected spectrophotometrically using a goat, anti‐human immunoglobulin conjugated with horseradish peroxidase. The db‐ELISA was compared to a standard competitive‐binding ELISA (cb‐ELISA). A competitive‐binding radioimmunoassay (cb‐RIA) was used as the reference assay for this study. The specificity of the cb‐ELISA was 50% and the positive predictive value was 64% when compared to the cb‐RIA. The specificity and positive predictive value of the db‐ELISA were both 97% when compared to the cb‐RIA. Based on titration studies, the sensitivity of the db‐ELISA is at least equal to the cb‐RIA. Based on the data generated in this study, the db‐ELISA appears to be an acceptable screening test for anti‐HBc.
Deceptively Low Morbidity from Failure to Practice Safe Blood Transfusion: An Analysis of Serious Blood Transfusion Errors
Authors: Murphy W. G. , McClelland D. B. L.
Published: March 2009

Abstract. Within a 24‐month period, 5 patients in a large teaching hospital were mistakenly transfused with blood that had been crossmatched for different patients. Each of the incidents was due to failure by ward staff to adhere to established safely procedures. Three incidents were entirely due to failure to make the standard checks comparing the identity details on the blood pack label with the patient identification details. The fourth incident was due to a combination of inadequate checking and incomplete patient identification procedure. The fifth incident was due to a series of errors that consisted of inadequate checking, plus putting the wrong patient's blood into the sample tube, plus misspelling of the patient's name on both sample tube and request form. There was no serious morbidity. All of these incidents took place when an unusual coincidence or contributing error lead to unmasked inadequate checking of blood unit against patient's identity by ward staff. The incidence of inadequate checking technique may be much higher than the incidence of erroneous transfusion events. It may be difficult to test the ability of an ‘improved’ transfusion procedure to prevent disasters from erroneous unit‐to‐patient matching, since assessment will need to include actual ‘worst case’ situations which should be rare.
Comparison of Platelet Activation and Membrane Glycoprotein Ib and IIb‐IIIa Expression after Filtration through Three Different Leukocyte Removal Filters
Authors: Bertolini Francesco , Rebulla Paolo , Porretti Laura , Sirchia Girolamo
Published: March 2009

We evaluated three filters used for leukocyte removal from platelet concentrates: Imugard IG 500, Pall PL100 and Sepacell PL‐10A. Filter performance, platelet activation and expression of membrane glycoproteins Ib and IIb‐IIIa were evaluated. Imugard, Pall and Sepacell showed median postfiltration in vitro platelet recoveries of 88, 84 and 80%, and total residual leukocyte counts of 16.1, 7.5 and 0.6 times 106/pool of 8 platelet concentrates, respectively. Mean platelet volume was reduced after filtration with all filters. Postfiltration values of glycoproteins Ib and IIb‐IIIa, and of activation markers GMP 140 and gp 53 were not significantly different from prefiltration values. Filtration through Imugard, Pall and Sepacell did not induce significant platelet activation or modifications of platelet membrane glycoproteins Ib and IIb‐IIIa.
Immune Hemolytic Anemia Associated with Biclonal Cold Autoagglutinins
Authors: Tschirhart Donald L. , Kunkel Lori , Shulman Ira A.
Published: March 2009

A 65‐year‐old man with bladder outlet obstruction due to prostatic hypertrophy was incidentally discovered to have cold‐antibody autoimmune hemolytic anemia (cold‐aggluthinin syndrome; CAS) due to autoanti‐I (titer 1,024 at 4°C and 64 at 30°C), and a biclonal gammopathy. Immunofixation electrophoresis of serum and a red blood cell eluate revealed the patient's autoantibody to be biclonal IgMx and IgAx. No underlying cause could be determined to explain the development of either the biclonal gammopathy or the CAS. To our knowledge, this is the first reported case of CAS associated with a biclonal gammopathy and biclonal cold autoagglutinins of the IgMx, IgAx type.
Neolacto (Type‐2 Chain)‐Sialoautoantigens Recognized by Human Cold Agglutinins
Authors: Roelcke D. , Hengge U. , Kirschfink M.
Published: March 2009

The antigens Fl and Vo, recognized by cold agglutinins, are protease‐resistant differentiation antigens like I and i antigens. In contrast to Ii, Fl and Vo are sialidase‐susceptible antigens like the protease‐resistant Gd antigens. Because I/i antigens are branched/linear type‐2 (poly‐N‐acetyllactosamine or neolacto series) chains which are cleaved from red cells by endo‐β‐galactosidase, the effect of the enzyme on Fl, Vo and Gd antigens was studied. Fl and Gd antigens, previously shown to be sialylated branched (Fl), and linear and branched (Gd) type‐2 sequences, were not inactivated on red cells by the enzyme, whereas Vo antigenicity was completely abolished. The data suggest that Vo antigen is expressed on sialylated linear poly‐N‐acetyllactosamine sequences.
Platelet Concentrates in an Additive Solution Prepared from Pooled Buffy Coats.
Authors: Eriksson Lars , Högman Claes F.
Published: March 2009

Abstract. A new method for the preparation of platelet concentrates (PCs) is described. The source material is buffy coat (BC), prepared after keeping standard CPD whole‐blood units at room temperature for 6–12 h, followed by centrifugation at 3,500 rpm for 10 min (first series) or 4,000 rpm for 12.5 min (second series). BC, separated from plasma and red cells, was kept at room temperature for a further 8–12 h without agitation. Pools of 6 (first series) and 4 (second series) BCs were prepared using a sterile docking device and suspended in a platelet‐additive solution (PAS) containing sodium/potassium chloride, citrate, phosphate, and mannitol. After gentle centrifugation, the platelet‐rich supernatant was expressed to and stored in one (first series) or two (second series) 1‐liter polyolefine (PL‐732) containers. In the first series, the total number of platelets was 316 ± 59 times 109 per PC (yield 65%). However, when the method was applied at a routine scale, the yield varied considerably and was shown to be strongly dependent on the hematocrit of the BCs. A number of steps were taken to standardize the technique which resulted in an improved yield (77.3 ± 8.7%) with 316 ± 52 times 109 platelets (mean ±SD, range 203–490, n = 134), obtained from 4 BC pools and lower leukocyte contamination than before, 18 ±17 times 106 per preparation (range 1–73, microscopic counting, n = 38). The storage medium consisted of a mixture of plasma and PAS. Standardization of the hematocrit and volume of the BCs is essential, both for improvement of the yield and for providing a sufficient amount of glucose and bicarbonate to the platelets throughout storage. The method is particularly suitable in combination with automated techniques for blood component preparation which include BC removal. Blood gases, pH and adenosine triphosphate were maintained at satisfactory levels; the release of platelet factor 4 and lactate dehydrogenase was not different from traditional PC preparation and storage. However, the content of glucose was exhausted in many PC units towards the end of a storage of 6 days. When whole blood and BC are stored for such a long period, and when the gas permeability of their containers is poor, it is important, in order to avoid early loss of swirling, that the surface‐to‐volume ratio and the gas permeability of the PC containers is sufficiently large to allow good gas exchange.
Flow‐Cytometric Method for Counting Very Low Numbers of Leukocytes in Platelet Products1
Authors: Dzik Walter H. , Ragosta Annette , Cusack William F.
Published: March 2009

Abstract. We provide a detailed description of a new method for counting very low numbers of leukocytes present in platelet products. The method uses fluorescent staining of leukocyte DNA using propidium iodide and flow cytometry. The assay is accurate over a range of leukocyte concentrations from 0.1 to 10 WBC/μl. We validated the method using specially designed dilution studies and applied it to the measurement of leukocyte content in leukocyte‐depleted platelet products prepared by apheresis or filtration. This method should prove useful in studies which address the clinical impact of leukocyte‐depleted platelet products.
Screening for Anti‐Human T‐Lymphotropic Virus Antibody in Blood Donors and Polytransfused Patients in Apulia (Italy)
Authors: de Stasio G. , Canavaggio M. , Rizzi L. , Lattanzio A. , Lancieri M. , D'Erasmo F. , Colapietro A. , Barbuti S. , Quarto M. , Liso V. , Specchia G. , Pastore G. , Schreiber L. , Lee H.
Published: March 2009

Abstract. A stringent procedure for the diagnosis of human T‐lymphotropic virus (HTLV) infection was applied to 1,732 volunteer blood donors, 401 patients with various hematological disorders and 78 individuals at high risk for HIV infection. It consisted of a viral lysate‐based screening assay (Abbott Laboratories, North Chicago, Ill., USA), and two confirmatory assays (Western blot and radioimmunoprecipitation assay). A confirmed positive sample had to react with at least two different HTLV gene products. Evidence of HTLV infection was not found in either blood donors or patients with hematological disorders. In fact, HTLV infection was only observed in 10 intravenous drug abusers or their sexual partners. Contrary to previous reports that claimed HTLV seroprevalences of between 0.3 and 8% in blood donors from Apulia (Italy), our data suggest that infection with this virus is principally restricted to intravenous drug abusers.
The Platelet Storage Capability of Different Plastic Containers
Authors: Wallvik Jonas , Åkerblom Olof
Published: March 2009

Abstract. Platelet concentrates (PC), prepared by platelet apheresis, were stored in four different types of blood bags. One of the bags, manufactured with a thinner PVC film than previously, was tested in three different bag volumes. From 25 donors a total number of 99 PC were prepared. Platelet numbers varied from 20 to 140 times 109 platelets per bag. The cell count, pH, pO2, pCO2 and lactate were determined initially and on days 1,3 and 5 of storage. In a separate test, the oxygen diffusion capacity of the bags was determined by oxidation of sodium sulfite in the presence of cobaltous chloride. The oxygen diffusion capacity found was 16 (PL 732, 300 ml), 13.5 (Teruflexa 800 ml), 11.5 (PL 1240, 400 ml), 10.6 (Teruflexa 600 ml), 9 (Teruflexa 400 ml) and 4 (PL 146, 300 ml) umol O2/h, respectively. For each bag type, the minimum and maximum platelet number stored with maintained pH levels (6.9–7.4) was defined. The maximum platelet number stored with maintained aerobic metabolism, correlated to the oxygen diffusion capacity of the bag, r = 0.998, p<0.001, n = 6; thus the maximum platelet number successfully stored for 5 days in each container can be predicted by determination of the oxygen diffusion capacity. In PC with a low platelet yield, pH values above 7.4 were observed after 1 and 3 days. When the results are compared with platelet yield data from routine blood banking, the optimal bags for platelet storage can be chosen. These conclusions must be further investigated in studies in vivo.
A Simple Protocol for the Screening and Preliminary Identification of Antibodies to Human IgA Using Unpurified Proteins in the Passive Haemagglutination Test
Authors: Hunt A. , Reed M.
Published: March 2009

Abstract.Experiments have shown that suitably diluted, unpurified IgA paraprotein sera can be substituted for chemically purified IgA proteins in the passive haemagglutination test for the detection of anti‐IgA antibodies. The substitution of unpurified IgA paraprotein was made possible by the enhanced serological activity obtained by pre‐heating the unpurified immunoglobulin preparations at 56°C for 60 min. Using a small screening panel of suitably coated tanned red cells, various population groups were investigated for the presence of anti‐IgA antibodies. Results were similar to those obtained by other workers using purified IgA preparations in the passive haemagglutination test.
A Simple Non‐Isotopic Method for the Quantitation of Red Cell‐Bound Immunoglobulin
Authors: Kumpel Belinda M.
Published: March 2009

Abstract. A method was developed for the estimation of the number of molecules of immunoglobulin bound to red cells. Solubilization of sensitized, washed red cells was followed by an enzyme‐linked immunosorbent assay of immunoglobulin concentration (SOL‐ELISA). This assay gave results that were comparable to isotopic methods, was reproducible, utilised inexpensive reagents and was relatively quick and simple to perform.
An Outbreak of Non‐A, Non‐B Hepatitis Associated with the Infusion of a Commercial Factor IX Complex during Cardiovascular Surgery1
Authors: Gerber A. R. , Englender S. J. , Selvey D. , Carlson J. F. , Matthews D. L. , Webster H. M. , Caldwell G. G.
Published: March 2009

Abstract. An outbreak of non‐A, non‐B hepatitis was recognized among cardiovascular surgical patients from one hospital in June 1985. Illness was found to be significantly associated with a commercial brand of factor IX complex given to patients because of intraoperative bleeding. A change in the commercial brand of factor IX stocked by the hospital pharmacy had occurred in October 1984 and coincided with the onset of the outbreak. A retrospective study of cardiovascular surgery patients identified 23 cases and 7 probable cases of non‐A, non‐B hepatitis among patients who had received infusions of brands A and B factor IX complex. Three cases were in brand A recipients and 27 were in brand B recipients. Respective brand‐specific attack rates were 5 and 42% (relative risk = 7.7; p<2 × l0‐5; χ2 test). Nineteen of 30 case patients (63%) were jaundiced, including 2 brand A recipients and 17 brand B recipients. Median peak serum aminotransferase was 690.5 IU (range 27–2,824). The incubation period for cases in brand B recipients was a median of 7 weeks (range 2–17 weeks).

Prevention of non‐A, non‐B hepatitis in this population requires adhering to strict indications for the use of clotting factor preparations and avoiding these products when at all possible. Heat treatment of clotting factor products may reduce the risk of viral disease transmission, but certain methods may not inactivate the causative agents of non‐A, non‐B hepatitis.
Erythrocyte Endogenous Proteinase Activity during Blood Bank Storage
Authors: de Angelis V. , de Matteis M. C. , Orazi B. M. , Santarossa L. , Toffola L. Della , Raineri A. , Vettore L.
Published: March 2009

Abstract. We studied proteolytic alterations of membrane proteins in ghosts derived from human red blood cells, preserved up to 35 days in the liquid state either as whole blood or with additive solution. The study was carried out by performing sodium dodecyl sulfate polyacrylamide gel electrophoresis of stromal proteins from erythrocytes, either previously treated with proteinase inhibitors or previously incubated in conditions promoting proteolysis. To differentiate the effect of erythrocyte from granulocyte proteinases, the investigation was also carried out in leukocyte‐free red cell preparations. The results show: (1) the effects of endogenous proteinases on membrane proteins derived from red cells stored under blood bank conditions; (2) a decrease of proteolytic effects in ghosts derived from red cells which have been submitted to a longer storage; (3) a relevant influence of the red cell resuspending medium before lysis on the time‐dependent onset and exhaustion of proteolysis in ghosts. The presence of increased proteolysis in ghosts could be regarded as a marker of molecular lesions induced in red cells by storage under blood bank conditions.
Possibility to Improve Preservation of Whole Blood by Leukocyte‐Depletion before Storage
Authors: Riedner C. , Heim M. U. , Mempel W. , Wilmanns W.
Published: March 2009

Abstract. In order to evaluate the influence of leukocytes in whole blood on the standard blood bank conditions for storage, we compared several parameters of red cells, leukocytes and platelets in leukocyte‐rich and leukocyte‐poor whole blood over a storage period of 5 weeks. The leukocyte‐depleted units were prepared by filtration with a leukocyte trapping filter (Sepacell R500A). Our data indicate improved storage conditions, made evident by significantly lower cell damage: smaller increases of elastase, β‐thromboglobulin and lactate dehydrogenase in leukocyte‐depleted blood. This improvement was achieved in a simple two‐bag system using only filtration, without addition of rejuvenating solutions. In addition, this procedure allows greater flexibility in the use of filtrated blood for transfusions.
In vitro Evaluation of Buffy‐Coat‐Derived Platelet Concentrates Stored in a Synthetic Medium1
Authors: Fijnheer R. , Veldman H.A. , van den Eertwegh A.J.M. , Gouwerok C.W.N. , Homburg C.H.E. , Boomgaard M.N. , de Korte D. , Roos D.
Published: March 2009

Abstract. Human blood platelets, prepared by the buffy‐coat method, were prepared and stored in different synthetic media. In a synthetic medium based on gluconate, acetate and citrate (GAC), the pH was 6.8 on day 6. This medium was chosen for further evaluation. The total platelet count and the leukocyte contamination were significantly lower in the platelet concentrates (PCs) prepared in GAC compared to PCs prepared in plasma. Platelets stored in plasma or in GAC were equally functional when tested for aggregation and adenosine triphosphate (ATP) secretion. Only stimuli that act through the arachidonic‐acid pathway induced a lower platelet response in GAC. Platelet morphology was quantified by measuring the difference in light transmission during stirring at different rates in an aggregometer; no significant differences for platelets stored in GAC as compared to plasma were observed. Activation of platelets was measured by binding of monoclonal antibodies (McAb) against the Gp IIb/IIIa complex and against activation‐dependent antigens (GMP 140 from the α‐granules and a 53‐kD glycoprotein from the lysosomal granules). There was no difference in binding of these McAb between platelets prepared and stored in plasma or GAC. We conclude that platelets prepared by the buffy‐coat method and stored in GAC have the same in vitro qualitities as platelets stored in plasma, except for the lower aggregation response by the archidonic‐acid pathway. This is probably due to an acetate‐induced decrease in intracellular pH.
A Study of the Effect of AB0 Incompatible Plasma in Platelet Concentrates Transfused to Bone Marrow Transplant Recipients
Authors: Shanwell A. , Ringdén O. , Wiechel B. , Rumin S. , Âkerblom O.
Published: March 2009

Abstract. Bone marrow transplant recipients at Huddinge Hospital have routinely received single‐donor platelet concentrates (PC) from blood group 0 donors. These PC contain approximately 350 ml plasma, which is incompatible with patients of group A, B and AB. In 27 patients transplanted with an ABO‐identical bone marrow, serological investigations have been performed every week after transplantation (median 6 weeks, range 3–14). Nine of 11 recipients with blood group A developed a positive direct antiglobulin test (DAT) after PC transfusions, while none of 15 patients of blood group 0 developed a positive DAT. Anti‐A could be eluted in DAT‐positive cases. In no case was there any clinical sign of hemolysis. Nor did recipients of groups A, B or AB (n = 34) require more red blood cells or PC transfusions compared to recipients of group 0 (n = 47).
Manipulating Red Cell Intra‐ and Extracellular pH by Washing1
Authors: Meryman H. T. , Hornblower M.
Published: March 2009

Abstract. Washing red cells with solutions containing no anions capable of entering the cells is known to result in the loss of intracellular chloride and a counterflow of OH‐ which raises intracellular and lowers extracellular pH. Elevating the intracellular pH improves the quality of the cells during 4°C storage. The extent to which intracellular pH can be raised and extracellular pH reduced depends not only on the inability of extracellular anions to enter the cells but also on the buffering capacity of the wash solution. Both intra‐ and extracellular pH can be manipulated by the judicious selection of anions or combinations of anions used in the wash solution.
Identification by Immunoblotting of the Erythrocyte Membrane Sialoglycoproteins That Carry the Vw and Mur Antigens
Authors: Herron Bob , Smith Graham A.
Published: March 2009

Abstract. Immunoblotting of the separated membrane components of MiI erythrocytes with anti‐Vw identified a band of Mr 40,000 with a mobility close to that of β‐sialoglycoprotein corresponding to the abnormal α‐sialoglycoprotein present in MiI cells. A comparison of results obtained when MiIII erythrocytes were immunoblotted with anti‐Mur, anti‐s and the monoclonal antibody R1.3, indicated that the Mur antigen is located on the abnormal δ‐sialoglycoprotein of Mr 36,000 in MiIII erythrocytes. Prior treatment of MiI erythrocytes with neuraminidase resulted in an increase in the intensity of staining of the anti‐Vw reactive component. This was consistent with the enhanced reactions observed in haemagglutination tests with neuraminidase‐treated erythrocytes. The mobility of the Vw component was reduced when erythrocytes were pre‐treated with low concentrations of neuraminidase but increased when higher concentrations of neuraminidase were used.
Comparison of Platelet‐Rich Plasma Collection Using the Haemonetics PCS and Baxter Autopheresis C
Authors: Anderson N.A.B. , Pamphilon D.H. , Tandy N.J. , Saunders J. , Fraser I.D.
Published: March 2009

Abstract. We have compared 118 platelet‐rich plasma donations collected using the Autopheresis‐C Platelet cellTM (Auto C) with 166 donations using the Haemonetics PCS. The median platelet yield from the Auto C was superior (2.51 vs. 1.54 × 1011) although collection times differed (60 vs./40min). There was greater variability in the platelet yield from the Auto C (0.45–5.6 VS./0.26–2.8 × 1011), but leucocyte contamination was not significantly different. After secondary processing, there was significantly less residual platelet‐poor plasma (272 vs. 369 ml). Platelet function assessed during 5 days of storage was satisfactory for both, although platelet aggregation responses to collagen and adenosine diphosphate were superior in the Auto C platelets.
Low Prevalence of Antibodies against Human Immunodeficiency Virus in Finnish Haemophiliacs
Authors: Rasi V. , Ikkala E. , Myllylä G. , Nevanlinna H. R.
Published: March 2009

Abstract. National yearly surveys were carried out between 1985 and 1989 to determine the prevalence of antibodies for human immunodeficiency virus (HIV) in Finnish patients with bleeding disorders. From 192 out of the 214 haemophiliacs (90%) tested, 2 patients were positive for anti‐HIV. No seropositivities were found after 1985. Fourteen out of 21 patients (67%) with type III von Willebrand's disease, and 7 out of 8 patients (88%) with factor XIII deficiency were tested with negative results. The low prevalence of anti‐HIV (0.94%; 2/213 tested), is mainly due to the self‐sufficiency for clotting factors, the low prevalence of HIV in the population, and the use of cryoprecipitate during the critical period.
Hepatitis C Virus Antibodies in High‐Risk Saudi Groups
Authors: Bahakim H. , Bakir T.M.F. , Arif M. , Ramia S.
Published: March 2009

Abstract. The recombinant‐based enzyme immunoassay developed by Ortho Diagnostic System for the detection of antibodies of hepatitis C virus (HCV) was used to determine the extent of exposure to HCV in healthy Saudi subjects (500 males; 260 females) without liver disease and with no history of percutaneous exposure to blood and in Saudis who are considered at high risk for contracting the disease: hemophiliacs (28), thalassemics (78), hemodialysis patients with renal failure (65) and patients with sexually transmitted diseases (STD) (220). The results show that HCV is endemic in the Saudi population with an overall frequency of 5.3% in healthy Saudi adults which is at least 5 times higher than what has been reported from Western Europe and the United States. Seropositivity rate in the high‐risk groups ranges from 15.9% in patients with STD to 78.6% in hemophiliacs. These data underscore the urgent need for routine anti‐HCV screening of blood donations in order to reduce the frequency of postransfusion non‐A, non‐B hepatitis and its sequelae in the Saudi population.
Alanine Aminotransferase, Gamma‐Glutamyltransferase, Antibodies to Hepatitis B Core Antigen and Antibodies to Hepatitis C Virus in Blood Donor Screening
Authors: Ebeling F.
Published: March 2009

Abstract. Alanine aminotransferase (ALT), γ‐glutamyl‐transferase and hepatitis B core antibodies were evaluated as donor markers in a prospective study of 685 open‐heart surgery patients. Of these three surrogate markers, only an ALT level greater than or equal to 2 SD above the log mean had a significant association with recipient non‐A, non‐B hepatitis (NANBH, p = 0.02). Antibodies to the hepatitis C virus (anti‐HCV) were detected by an enzyme immunoassay in 7 of the 136 units transfused to the 11 NANBH patients and 29 of 3,650 not associated with hepatitis (p < 0.001). Calculated from this subgroup of donors, the anti‐HCV test would have a 15.6% positive predictive value with 0.92% donor loss and thus is superior as a primary screening marker to all the three surrogate tests. The predictive value could be substantially increased by subsequent ALT testing or by the use of a recombinant immunoblot anti‐HCV assay.
Enhanced Detection of HIV‐1 Sequences Using Polymerase Chain Reaction and a Liquid Hybridization Technique
Authors: Bruisten S. M. , Koppelman M. H. G. M. , van der Poel C. L. , Huisman J. G.
Published: March 2009

Abstract. In this report we describe a sensitive HIV‐1 detection method which is applicable for confirmation testing of donors whose blood sample gives indeterminate viral‐serology results. The method involves performing a polymerase chain reaction (PCR) and detecting the generated fragments using liquid hybridization and gel retardation. We found that it is as specific as blotting on a filter and hybridization with an internal probe but at least tenfold more sensitive. After applying it on DNA samples of a panel of 11 persistent indeterminate anti‐p24 gag‐reactive donors, none was found to be PCR positive. Considering other negative virological and biochemical test results and case‐historical data, these donors are not likely to be HIV‐1 infected.
Significance of Human T‐Lymphotropic Virus Type I Indeterminant Serological Findings among Healthy Individuals
Authors: Lipka James J. , Young Karen K. Y. , Kwok Shirley Y. , Reyes Gregory R. , Sninsky John J. , Foung Steven K. H.
Published: March 2009

Abstract. Follow‐up studies on 67 blood donors with indeterminant serological findings for human T‐lymphotropic virus (HTLV) type I by standard immunoassays showed no evidence of infection by polymerase chain reaction analysis for HTLV‐I or HTLV‐II nucleic acids or by antibody reactivity to a unique HTLV‐I recombinant envelope protein, MTA‐4. Among HTLV‐I‐ or ‐II‐infected individuals, a history of blood transfusion, past residence in established HTLV‐I endemic areas or some association with intravenous drug use were common. In contrast, 85% of indeterminant cases had none of these risk factors. These observations suggest that healthy individuals with indeterminant serology for HTLV‐I should not require additional studies.
Safety of Intravenous Immunoglobulin Preparations: A Prospective Multicenter Study to Exclude the Risk of Non‐A, Non‐B Hepatitis
Authors: Imbach P. , Perret B.A. , Babington R. , Kaminski K. , Morell A. , Heiniger H.J.
Published: March 2009

Abstract. The risk of non‐A, non‐B hepatitis transmission by an intravenous immunoglobulin (IVIG) preparation was assessed in a prospective multicenter trial in 68 patients with primary immunodeficiency disorders (40 children or adolescents and 28 adults). During the 4‐week prestudy evaluation period the clinical examinations and liver function tests including alanine aminotransferase, aspartate aminotransferase, γ‐glutamyl transpeptidase, alkaline phosphatase, and bilirubin were normal in all patients. The treatment consisted of three infusions of 200 mg IVIG (pH 4; pepsin procedure) per kilogram body weight at 2‐week intervals. During the observation period of 24 weeks following the first infusion of the study IVIG, the patients were monitored at regular time intervals. No clinical and laboratory signs of hepatitis or liver dysfunction were noticed. All patients completed the study. In 5 patients, one isolated alanine aminotransferase value and in another patient one γ‐glutamyl transpeptidase value were moderately elevated, but always below 2.5 times the upper limit of the reference range. Similar isolated and transient elevations were observed for aspartate aminotransferase and alkaline phosphatase. It was concluded that the IVIG preparation did not transmit non‐A, non‐B hepatitis or other viral liver diseases.
Relative Functional Binding Activity of IgGl and IgG3 Anti‐D in IgG Preparations
Authors: Gorick B. D. , Hughes‐Jones N. C.
Published: March 2009

Abstract. The intention to replace polyclonal IgG anti‐D with human monoclonal antibody in the prophylaxis of haemolytic disease of the newborn requires knowledge concerning the relative content of IgGl and IgG3 anti‐D in prophylactic IgG preparations that are in present use. This has been carried out using a functional assay in which the absolute amount of IgGl and IgG3 anti‐D present on red cells was determined after incubation with IgG preparations. The assay was carried out by flow cytometry on 17 samples; expressed as a percentage of the total, the average value for the amount of IgG3 anti‐D on the cells was 8% (range 1–18%). Similar measurements were also made on the anti‐D present in 18 samples of antisera; IgG3 anti‐D formed a larger fraction of the total, the average value being 17% (range 0–60%) confirming previously reported estimates. It is suggested that some of the low values found for IgG3 in IgG preparations may be due do preferential loss during production.
Flow‐Cytometric Analysis of Erythrocytic Blood Group A Antigen Density Profile
Authors: Berneman Zwi N. , van Bockstaele Dirk R. , Uyttenbroeck Wim M. , van Zaelen Christiane , Cole‐Dergent Joséphine , Muylle Ludo , Peetermans Marc E.
Published: March 2009

Abstract. Blood group A antigen density on red blood cells (RBC) was studied using flow cytometry (FCM) and fluoresceinated polyclonal and monoclonal IgG anti‐A antisera. Agglutination was a problem, which could only be solved by prefixation of the RBC with glutaraldehyde or formaldehyde. However, this treatment resulted in a significant reduction of the number of antigen sites as compared to the native (i.e. nonfixed) RBC. Two major new findings came out of this study: (1) A antigen density on native RBC seems to be higher than previously recognized, and (2) A antigen density distribution is probably non‐Gaussian. The absolute number of A antigen sites was determined, using a human polyclonal IgG antiserum and commercially available absolute fluorescence standards. The site numbers on fixed RBC were comparable to those found by earlier radioimmunological studies (× 106/RBC): A1, 1.07 ± 0.28; A2,0.21 ± 0.09; A1B, 0.79 ± 0.26 sites (mean ± SD). The values found for native RBC were considerably higher (× 106/RBC): A1, 2.86 ± 0.95; A2,0.47 ± 0.29; A1B, 1.98 ± 0.58 sites (mean ± SD). With the 1 monoclonal and the 3 polyclonal antisera used in this study, and in contrast to Rh D, the erythrocytic A antigen density distribution of a given sample is highly asymmetrical. This non‐Gaussian distribution profile does not seem to be affected by such factors as antibody heterogeneity, variability in antibody fluoresceination range, RBC density and reticulocyte content. This suggests that the asymmetrical A antigen distribution may be an intrinsic property of the RBC population.
Interviews with Anti‐HIV‐Positive Individuals Detected through the Systematic Screening of Blood Donations: Consequences on Predonation Medical Interview
Authors: Lefrère Jean‐Jacques , Elghouzzi Marie‐Hélène , Paquez François , N'Dalla Jacob , Nubel Luc
Published: March 2009

This study is based upon interviews with 74 individuals found to be human immunodeficiency virus (HIV)‐seropositive through the screening of blood donations between January 1988 and December 1990. The donation history and the risk factor of HIV infection were established. Questions about the use of blood donation as a diagnostic test and on the notion of a predonation medical interview evoking the risk factor were asked. The majority of the individuals had a risk factor of HIV infection and had given their blood for serological testing. This data can help to adapt the predonation medical interview to the present epidemiological context of HIV infection. The improvement of this interview will contribute to the decrease of the residual transfusional risk of HIV infection.
A Murine Monoclonal Antibody Directed against the Gerbich 3 Blood Group Antigen
Authors: Loirat M.J. , Gourbil A. , Frioux Y. , Midler J. Y. , Blanchard D.
Published: March 2009

A murine monoclonal antibody (NaM19–3C4, IgG1, Kappa) was produced from splenocytes of mice immunized with red blood cells. The antibody agglutinated untreated Ge:2,3,4 and Ge:‐2,3,4 erythrocytes in indirect antiglobulin test but failed to agglutinate trypsin‐treated cells. Gerbich‐negative erythrocyte of the Leach‐ (Ge:‐2,‐3,‐4) and of the Gerbich‐ (Ge:‐2,‐3,4) types were not recognized by the antibody. Immunoblotting experiments showed that the antibody bound to glycophorins C and D from control erythrocytes and to the abnormal glycophorin C identified in the Gerbich‐negative cells of the Yussef type (Ge:‐2,3,4). No binding to the altered glycophorin C from Ge:‐2,‐3,4 erythrocytes was observed, indicating that the antibody specifically recognized the Ge:3 epitope localized within residues 40–50 of glycophorin C.
First Autoclave‐Sterilized Platelet‐Additive Solution Containing Glucose with a Physiological pH for the Preparation of Plasma‐Poor Platelet Concentrates
Authors: Shimizu T. , Shibata K. , Kora S.
Published: March 2009

The glucose‐free platelet‐additive solution (termed AR solution), developed by Adams and Rock [Transfusion 1988;28:217—220], was modified by adding glucose as an energy substrate for platelets and maltose to prevent platelet lysis and by replacing sodium gluconate with sodium phosphate for better pH maintenance. The new platelet‐additive solution (termed Seto solution) contained 90 mM NaCl, 5mM KCl, 3mM MgCl2, 17 mM tri‐sodium citrate, 4.9 mM NaH2PO4, 20.1 mM Na2HPO4, 23 mM sodium acetate, 28.8 mM maltose, and 23.5 mM glucose with a pH of 7.4. The solution was sterilized by autoclaving in plastic bags in nitrogen to prevent glucose caramelization at high pH. Plasma‐poor platelet concentrates prepared by adding Seto solution to the pelleted platelet buttons were stored in a LE‐2 polyolefin bag at 22 °C with constant agitation for 5 days. The platelets suspended in Seto solution maintained oxygen consumption at a rate of 1.1 nmol/min/109 platelets after 5‐day storage, with glucose consumption and lactate production rates of 0.5 ± 0.2 and 1.2 ± 0.2 nmol/min/109 platelets, respectively. This resulted in a final mean pH of 7.0. Those suspended in AR solution ceased glycolysis within 3 days because residual plasma glucose had been consumed. This was associated with decreases in percent hypotonic shock response and aggregation induced by adenosine diphosphate and collagen. Lactate dehydrogenase discharge in AR solution was 5 and 8 times higher at day 3 and day 5, respectively, than that of Seto solution. Morphologically, there were no ballooned platelets after storage in Seto solution. Platelets stored in Seto solution were nearly equal to those stored in plasma. Conclusions: Steam‐sterilized Seto‐additive solution caused better storage of platelets for 5 days than AR solution, suggesting that glucose is important for optimal storage. Phosphate acts as an effective buffer and maltose stabilizes platelet volumes.
Quantitation of the Human Component C4: Definition of C4 Q0 Alleles and C4A Duplications
Authors: Rebmann Vera , Doxiadis Mas , Kubens Britta S. , Grosse‐Wilde Hans
Published: March 2009

We determined the C4 plasma concentrations of 48 genotypically C4A‐ and C4B‐defined unrelated individuals from 34 families with a total of 196 members by an enzyme‐linked immunosorbent assay using Rg:1,2 (C4A) and Ch:1 (C4B) specific monoclonal antibodies. The results obtained allowed the establishment of rules for the detection of C4 Q0 alleles in the heterozygous form and of C4A gene duplications. In the present study seven homoduplications of the C4A 3 allotype were defined which had not been detected by allotyping. This procedure allows the simple, reliable, and quick determination of Rg:1,2 and Ch:l plasma levels which are not influenced by daily rhythms of C4 production.
Studies in Red Blood Cell Preservation: 5. Determining the Limiting Concentrations of NH4Cl and Na2HPO4 Needed to Maintain Red Blood Cell ATP during Storage
Authors: Dumaswala U.J. , Oreskovic R. T. , Petrosky T. L. , Greenwalt T.J.
Published: March 2009

The purpose of the present study was to define the lowest concentrations of ammonium (NH4+) and phosphate (Pi) in an experimental additive solution (EAS) that would support suitable red blood cell (RBC) ATP levels and other in vitro characteristics for at least 84 days. It was determined that ATP maintenance was dependent upon both NH4+ and Pi concentrations. RBCs stored for 84 days in additive solutions containing 10 mM NH4+ and 0,15, 25 and 40 mM Pi had ATP values averaging 1.87, 2.49, 2.70 and 2.65 μmol/g Hb, respectively. The shedding of exocytic hemoglobin‐containing vesicles and percent hemolysis were significantly (p < 0.001) elevated in the preservative containing 40 mM Pi. These data suggest that an EAS containing 10 mM NH4+ and 15 mM Pi would be optimal for storing RBCs up to 84 days. The extended storage would be particularly advantageous for autologous transfusion programs.
A Study of Autologous Blood Collected after Joint Replacement Surgery
Authors: Robbins G. , Grech H. , Howes K.
Published: March 2009

Blood from the surgical drains of 11 patients undergoing joint replacement was collected in the Solcotrans Orthopaedic autologous transfusion device and analysed for microparticulate matter before and after micro‐aggregate filtration and for its effect on the coagulation of paired venous blood samples. An average of 165 ml (range 0–260 ml) was collected into the Solcotrans during the first hour. Using a Coulter Counter Zm particle counter, particulate matter of diameter 10—20 μm was found in only 2 of 10 collections at an average concentration of 33 × 103/l. All units contained acoagulable blood [kaolin partial thromboplastin time (KPTT) > 600 s] but when mixed with paired post‐operative venous samples exhibited the ability to shorten the KPTT by an average of 4.3 s inspite of the marked dilutional effect of mixing. Retransfusion of blood collected in the Solcotrans Orthopaedic device appears to be a suitable method to supplement or substitute pre‐deposit and reduce exposure to homologous blood. Given the low incidence and concentration of microparticles detected, retransfusion of shed blood by this method is unlikely to cause significant pulmonary vascular occlusion resulting directly from deposition of microparticles.
Replacement Therapy with Virus—Inactivated Plasma Concentrates in von Willebrand Disease
Authors: Rodeghiero Francesco , Castaman Giancarlo , Meyer Dominique , Mannucci Pier Mannuccio
Published: March 2009

In von Willebrand disease, the goal of treatment is to correct the two laboratory hallmarks of abnormal hemostasis, i.e. the deficiency of factor VIII (F VIII) and the prolonged bleeding time (BT). Since desmopressin (DDAVP) is able to achieve both these goals in the majority of patients, it is the treatment of choice. Some patients, however, are not responsive or become refractory to DDAVP. In these, blood products maintain an important therapeutic role, and there is a need to assess the efficacy of the recently available virus‐inactivated plasma concentrates, which contain both F VIII and von Willebrand factor and carry a low risk of transmitting blood‐borne viruses. Our survey of the data reported in the literature indicates that all available concentrates are similarly effective in attaining high and sustained levels of FVIII after infusion. Although they often shorten or normalize the prolonged BT, that effect is less uniform. Since concentrates appear efficacious in the majority of clinical situations that require the use of blood products, they should be preferred, because of their greater safety, to cryoprecipitate produced by blood banks, which cannot be virus inactivated.
Early Antihepatitis C Virus Response with Second—Generation C200/C22 ELISA
Authors: Van der Poel C. L. , Bresters D. , Reesink H. W. , Plaisier A. A. D. , Schaasberg W. , Leentvaar‐Kuypers A. , Choo Q.‐L. , Quan S. , Polito A. , Houghton M. , Kuo G. , Lelie P. N. , Cuypers H.T.M.
Published: March 2009

Detection of early antibody to hepatitis C virus (HCV) by a new second‐generation C200/C22 anti‐HCV enzyme‐linked immunosorbent assay (ELISA) and a four‐antigen recombinant immunoblot assay (4‐RIBA) was compared with the first‐generation anti‐HCV C100 ELISA using sequential serum samples of 9 recipients who were infected with HCV, as detected by polymerase chain reaction after transfusion of blood products. Within 26 weeks after transfusion, 9/9 (100%) recipients seroconverted with C200/22 ELISA, and 6/9 (67%) seroconverted with C100 ELISA. Compared with C100 ELISA, C200/C22 ELISA seroconversion occurred simultaneously in 3 cases, 5–6 weeks earlier in 3 other cases, and 20 weeks earlier in 1 case. Seven of 9 (78%) recipients became positive, and 2/9 (22%) became indeterminate with 4‐RIBA. In 8 cases with clinical posttransfusion hepatitis non‐A, non‐B (PTH‐NANB), anti‐HCV C200/C22 ELISA seroconversion occurred 2–17 (mean 6) weeks after the onset of hepatitis. In 6 cases of PTH‐NANB, anti‐HCV C100 ELISA seroconversion occurred 2–26 (mean 9) weeks after the onset of hepatitis. It is concluded that the second‐generation C200/C22 ELISA is more sensitive than the C100 ELISA for the detection of antibody during early HCV infection. Indeterminate 4‐RIBA results are found in the early phase of HCV infection.
Enhanced Sensitivity of a Second Generation ELISA for Antibody to Hepatitis C Virus
Authors: Bresters D. , Cuypers H. T. M. , Reesink H.W. , Schaasberg W. P. , van der Poet C. L. , Mauser‐Bunschoten E. P. , Houghton M. , Choo Q.‐L. , Kuo G. , Lesniewski R. , Troonen H. , Lelie P. N.
Published: March 2009

A second generation ELISA for combined detection of antibodies to three hepatitis C virus (HCV) recombinant proteins, i.e. C100, C33c and core, was compared with a first generation anti‐HCV ELISA in which only antibodies to C100 are detected. The results of the ELISAs were evaluated in 225 haemophilia patients (panel A) and 44 patients with non‐A, non‐B (NANB) hepatitis (panel B). HCV infection was established by cDNA‐polymerase chain reaction (PCR) (in panel B only) and by studying the anti‐HCV reaction patterns in 4 separate ELISAs for detection of antibodies to the recombinant proteins C100, C33c, core and a combination of two synthetic peptides sp67/65 derived from the C100 region. The sensitivity for the detection of HCV infection had increased from 0.92 [95% confidence interval (CI): 0.87–0.95] to 1.00 (95% CI: 0.98–1.00) in haemophiliacs and from 0.84 (95% CI: 0.66–0.95) to 1.00 (95% CI: 0.89–1.00) in NANB hepatitis patients when the second generation ELISA was used instead of the first generation ELISA. Concurrently the chance of a false negative result was reduced in panel A and B from 0.37 to 0 and from 0.28 to 0, respectively. Analysis of anti‐HCV reaction patterns revealed that 172 of 206 (83.5%) anti‐HCV ELISA‐reactive haemophilia patients had antibodies to all 4 antigens tested. In the NANB hepatitis patients 18 of 31 (58.1%) anti‐HCV ELISA‐reactive subjects reacted with 4 antigens. In the PCR tested panel of NANB hepatitis patients 2 subjects who showed antibody reactivity to only one antigen and 5 patients with reactivity to 2 antigens were PCR‐positive. On the other hand, 1 NANB hepatitis patient showed antibody reactivities to all 4 antigens tested, but was PCR‐negative. Antibodies to core were the most prevalent in the two populations studied. We conclude that the introduction of the second generation anti‐HCV ELISA will significantly reduce the likelihood of unrecognized HCV infection.
Hepatitis C Viraemia in United Kingdom Blood Donors: A Multicentre Study
Authors: Garson J. A. , Clewley J. P. , Simmonds P. , Zhang L. Q. , Mori J. , Ring C. , Follett E. A. C. , Dow B. C. , Martin S. , Gunson H.
Published: March 2009

Of 10,633 blood donations tested in three regional blood transfusion centres with two commercial first generation screening assays for antibodies to the hepatitis C virus (HCV), 65 (0.61%) were found to be repeatedly reactive in one or both assays. Five of the 65 were confirmed positive by recombinant immunoblot assay (Ortho RIBA‐2) and a further 4 were judged indeterminate. All 5 RIBA‐2 positive donations and 1 of the 4 RIBA‐2 indeterminates were shown to be viraemic by HCV‐RNA polymerase chain reaction (PCR) assays performed at three independent reference laboratories. The remaining 56 screen test reactive donations proved negative by RIBA‐2 and, with 1 exception, negative by PCR. We conclude that while first generation anti‐HCV screening assays generate a high proportion of false reactions when screening low prevalence populations, results of the RIBA‐2 confirmatory test correlate well with PCR findings and thus indirectly with both hepatitis C viraemia and infectivity.