bluuebunny/crossref_metadata_2025 huggingface dataset:
Review of the Hepatitis A Epidemics in Hemophiliacs in Europe
Authors: Vermylen Jos , Peerlinck Kathelijne
Published: March 2009

A recent increased incidence of hepatitis A in the hemophilia populations of Italy, Germany, Belgium and Ireland was studied. The first well‐documented outbreak occurred in Italy, where 51 patients with severe hemophilia contracted hepatitis A between the years 1989 and 1992. A case‐control study by Mannucci et al. of the first 29 consecutive cases of hemophilia patients with jaundice led to the conclusion that hepatitis A was transmitted to Italian hemophilia patients by solvent‐detergent‐treated, ion‐exchange‐purified concentrate. In a retrospective serologic study of the German epidemic, Brakmann et al. isolated 18 seroconversions in a study group of 195 hemophilia patients over a 5‐year period. Although several types of concentrate were used in Germany during this time, all but 1 infected patient received large doses of solvent‐detergent, ion‐exchange product. It was revealed that the 1 exception had a brother with hemophilia who had received solvent‐detergent‐prepared concentrate and had developed hepatitis A. Transmission to the study patient was therefore attributed to household contact. The Belgian experience corroborated the findings of the Italian and German investigators: several batches of factor VIII, prepared from plasma of Belgian donors in Lille, France, and Vienna, Austria, appeared to be associated with hepatitis A infection, mainly in hemophilia patients who had received large infusions. An interesting observation in the later (1993) case‐control portion of the Belgian study was that hepatitis A virus antibodies were less prevalent in hemophilia patients than in age‐matched controls. A possible explanation for this phenomenon is that the hemophilia patients had been passively protected through the immunoglobulin contained in the cryoprecipitate they had received in early treatments, and that they had lost this immunity as a result of their more recent treatment with solvent‐detergent, ion‐exchange concentrate, which contains almost no immunoglobulin. Solvent‐detergent, ion‐exchange product was likewise implicated in the Irish epidemic of hepatitis A in which the highest incidence was observed: 30 of 215 Irish hemophilia patients became infected between May and December 1992. In Ireland, geographic clustering was also a factor, since all of the plasma used to prepare the 3 implicated lots of factor VIII originated in donors from the same geographic area. It was concluded that the solvent‐detergent, ion‐exchange chromatography‐purified method of factor VIII preparation was responsible for the recent epidemics of hepatitis A among hemophilia populations in Europe.
FDA Perspective on and Response to the Risk of Hepatitis A from Blood Products
Authors: Fricke William
Published: March 2009

The clinical and laboratory data available on hepatitis A virus infections reported in Europe were reviewed by the Blood Products Advisory Committee of the Food and Drug Administration. The panel's consensus was that the clotting factor concentrates currently available in the United States are safe with respect to viral diseases, hepatitis A transmissions do not appear to be a significant health problem, and there is a paucity of epidemiologic data on hepatitis A in patients with hemophilia in the United States.
Sensitivity and Specificity of Three Third‐Generation Anti‐Hepatitis C Virus ELISAs
Authors: Vrielink H. , Zaaijer H.L. , Reesink H.W. , van der Poel C.L. , Cuypers H.T.M. , Lelie P.N.
Published: March 2009

Three commercially available 3rd‐generation anti‐HCV ELISAs (Abbott, Murex and Ortho) were evaluated in various serum panels: (A) blood donor samples (n = 403) with 1st‐ or 2nd‐generation anti‐HCV ELISA (various manufacturers) positive test results; (B) non‐A, non‐B hepatitis patients (n = 212); (C) multitransfused patients (n = 253); (D) serial dilutions of HCV confirmed (RIBA and PCR) positive blood donors (n = 24), and (E) first‐time blood donors (n = 1,055). All samples of panels A, B and C were tested in PCR and RIBA‐2. In panels A, B and C, 398 samples were HCV PCR positive: all were detected by Abbott and Ortho, and 397 (99.7%) by Murex. The sample missed by the Murex ELISA showed an isolated anti‐C33c reactivity in RIBA‐2. In panels A–C, 442 samples were RIBA‐2 positive and all were detected by the 3 tests. With Probit analysis on results of panel D, no significant difference in sensitivity was observed between the 3 evaluated ELISAs. Specificities of Abbott, Murex and Ortho in 1,055 blood donors were 99.7, 99.3 and 99.9%, respectively (NS, χ2). We conclude that the sensitivity and specificity of the 3 ELISAs are comparable although the C33c antigen in the Murex VK47 test should be improved.
Hepatitis C Virus Genotypes among Blood Donors and Their Recipients in Iceland Determined by the Polymerase Chain Reaction
Authors: Löve Arthur , Smáradóttir Agnes , Thorsteinsson Sigurdur B. , Stanzeit Barbara , Widell Anders
Published: March 2009

Eight antibody‐positive individuals were detected among 12,000 blood donations during the first year of screening blood donors for hepatitis C virus (HCV) antibodies in Iceland. All 8 were found to have a history of intravenous drug abuse. Six of these 8 individuals had previously donated blood to 27 patients who could be traced and examined for HCV infection. The great majority (23/27, 85%) of the recipients had demonstrable HCV antibodies. Furthermore, RNA analysis with the polymerase chain reaction showed that all patients with HCV antibodies had HCV RNA in their serum and in one hemodialysis patient without HCV antibodies viral RNA could be demonstrated. Genotyping of the HCV strains showed that the genotype of the donor was also identified in all but one of the infected recipients of his/her blood or blood products. This study, therefore, substantiates high infectivity of the HCV by blood or blood factor donation and shows that viremic HCV antibody‐negative individuals exist.
A Novel Common Kell Antigen, TOU, and Its Spatial Relationship to Other Kell Antigens
Authors: Jones J. , Reid M.E. , Oyen R. , Harris T. , Moscarelli S. , Co S. , Leger R. , Beal C. , Cardillo K.
Published: March 2009

A 47‐year‐old native American (TOU) was admitted to hospital for hip surgery. His serum agglutinated all red blood cells (RBCs) tested except Ko and DTT‐treated RBCs and was weakly reactive with RBCs known to have a weak expression of Kell antigens, namely Kmod, McLeod, Kp(a+b‐) (KEL:3,‐4) and K:‐13 (KEL:‐13) phenotypes. RBCs from three siblings, a son and a daughter were incompatible with TOU's antibody. TOU's RBCs had the common Kell phenotype: K‐ k+ Kp(a‐b+c‐) Ku+ Js(a‐b+) Ul(a‐) K:11, ‐17 K:14,‐24 K:12,13,18,19,22,‐23 (KEL:‐1,2,‐3,4,5,‐6,7,‐10,11,12,13,14,‐17,18,19,‐21,22,‐23,‐24). Since TOU's RBCs were not agglutinated by an unidentified Kell‐related antibody (IAN), tests were performed to show that TOU and IAN were mutually compatible. IAN is a Latino female hospitalised for a hysterectomy. The TOU antigen was shown to be located on the Kell glycoprotein by a monoclonal antibody immobilisation of erythrocyte antigen (MAIEA) assay. The unique pattern of reactivity obtained with TOU and IAN antibodies using this assay indicated the TOU epitope to be in an area remote from other Kell antigens, namely K, k, Kpa, Kpb, Kpc, Ku, Jsa, Jsb, Ula, K11, K12, K13, K14, Wka, K18, K19, K22 and K24 (KEL1, KEL2, KEL3, KEL4, KEL5, KEL6, KEL7, KEL11, KEL12, KEL13, KEL14, KEL17, KEL18, KEL19, KEL21, KEL22 and KEL24) but close to the low‐incidence antigen K23 (KEL23). Investigation of antibodies to previously identified antigens on the Kell glycoprotein by MAIEA using the mouse monoclonal antibodies BRIC18, BRIC68, BRIC107 and BRIC203 has identified six patterns of reactivity and has provided evidence for Kpc being located in the same region as Kpa and Kpb.
No Detectable Alterations in Immunogenicity following Terminal Severe Dry‐Heat Treatment of High‐Purity Factor VIII (Liberate) and Factor IX (HP9) Concentrates
Authors: MacGregor I. R. , McLaughlin L. E. , MacGregor M. C. , Prowse C. V. , Pepper D. S.
Published: March 2009

We used monoclonal antibody ELISAs, antigen molecular size distribution, competition ELISA and neonatal mouse immune tolerance methods to detect potential neoantigen formation and increased immunogenicity following severe dry‐heat treatment of high‐purity factor VIII (Liberate) and factor IX concentrates. To provide positive controls, concentrates were heated in solution (70°C for 2 h) to produce denaturation on purpose. The competition ELISA applied to factor IX proved particularly useful for quantifying differences between the positive control and the dry‐heated/unheated concentrates. None of the test systems employed by us indicated any detectable neoantigen formation or any alteration in immunogenicity following terminal severe dry‐heat treatment of the high‐purity concentrates, and this finding is supported by clinical experience so far.
An Enzyme‐Linked Immunosorbent Assay for the Quantitative Determination of Anti‐D in Plasma Samples and Immunoglobulin Preparations
Authors: Hirvonen M. , Tervonen S. , Pirkola A. , Sievers G.
Published: March 2009

Here we present an enzyme‐linked immunosorbent assay (ELISA) on microtiter plates for the quantitative determination of anti‐D. This method is based on the solubilization of red blood cells sensitized with anti‐D and the subsequent measurement of immunoglobulin G by ELISA. The measuring range of the assay is 40–5,000 IU/ml and the lowest quantifiable concentration in plasma is 0.5 IU/ml. The interassay relative standard deviation for concentrations above 130 IU/ml ranges from 3.2 to 8.1% and below 50 IU/ml from 10.5 to 19.7%. Comparison of ELISA with automated hemagglutination shows that the results of the two assays correlate well: r = 0.992, n = 26. The assay was validated for donor plasma samples and anti‐D immunoglobulin preparations and it can also be used in assessing the severity of Rh (D) hemolytic disease during pregnancy.
DNA‐Based Typing of Human Platelet Antigen Systems by Polymerase Chain Reaction‐Single‐Strand Conformation Polymorphism Method
Authors: Fujiwara Koki , Tokunaga Katsushi , Isa Kazumi , Miyamoto Masaki , Wang Li , Akaza Tatsuya , Tadokoro Kenji , Shibata Yoichi , Juji Takeo
Published: March 2009

Polymerase chain reaction‐single‐strand conformation polymorphism (PCR‐SSCP) method has been established to discriminate genotypes for the human platelet antigen (HPA) systems HPA‐1, HPA‐2, HPA‐3, HPA‐4, and HPA‐5. Gene fragments which contain polymorphic sequences corresponding to the HPA‐1, HPA‐2, HPA‐3, HPA‐4, and HPA‐5 systems were PCR‐amplified with specific primers. The amplified DNA was denatured and subjected to non‐denaturing polyacrylamide gel electrophoresis followed by silver staining. The results obtained by the PCR‐SSCP method were in good agreement with those of the allotypes determined by serological typing. Furthermore, the results agreed with those obtained by other DNA‐based typing methods such as PCR‐allele‐specific restriction enzyme analysis and PCR‐sequence‐specific primer. These results indicate that PCR‐SSCP is a simple and sensitive method for determining HPA genotypes and identifying unknown polymorphisms.
Pharmacokinetics of Single and Multiple Infusions of 5S Intravenous Immunoglobulin
Authors: Theobald Klaus , Högy Barbara
Published: March 2009

A prospective, open and uncontrolled trial was performed to investigate the pharmacokinetics of a 5S intravenous immunoglobulin (IVIG) after single and multiple infusions. Twelve healthy volunteers received a 5S‐IVIG enriched in antibodies against Staphylococcus areus α‐toxin (aSAT) either as a single infusion or as three infusions in daily intervals. The fate of the injected immunoglobulin was observed by serial measurements of the serum level of aSAT titres. The 5S‐IVIG had a half‐life with a median of 0.24 (distribution) and 2.0 (terminal) days, respectively. The pharmacokinetic parameters did not differ with dose groups or number of applications. The study medication was well tolerated.
Improved Detection of Anti‐HCV in Post‐Transfusion Hepatitis by a Third‐Generation ELISA
Authors: Barrera J. M. , Francis B. , Ercilla G. , Nelles M. , Achord D. , Darner J. , Lee S. R.
Published: March 2009

The sensitivity of ORTHO™ HCV 3.0 ELISA Test System (ELISA 3) for the detection of anti‐HCV was compared with the second‐generation ELISA, ORTHO HCV 2.0 ELISA Test System (ELISA 2). ELISA 3 differs from ELISA 2 in that it incorporates the HCV recombinant antigen NS5, in addition to recombinant antigens derived from the NS3, NS4 and core regions of the HCV genome. Specimens tested consisted of serial bleeds obtained from 21 individuals undergoing seroconversion following acquisition of post‐transfusion HCV infection. ELISA 3 demonstrated significantly greater sensitivity than ELISA 2, detecting seroconversion earlier in 24% (5/21) of cases. Although one of these cases appeared to represent early seroconversion to NS5, most of the improved sensitivity of ELISA 3 appeared to derive from increased detectability of anti‐c33c.
Transmission of Hepatitis C Virus by Anti‐HCV‐Negative Blood Transfusion: Case Report
Authors: Vrielink H. , vander Poel C.L. , Reesink H.W. , Zaaijer H.L. , Lelie P.N.
Published: March 2009

Acute posttransfusion hepatitis C was reported in a recipient of 3 units of red cells. The recipient became acutely icteric 6 weeks after transfusion, and HCV infection was diagnosed. Stored serum samples of the 3 implicated donations, which were negative with ELISA‐2, were retested by PCR and 3rd‐generation antibody tests. One implicated donation was PCR positive, but anti‐HCV negative. Both other donations were negative in all tests. The donor was recalled to the Blood Bank 13 weeks after the implicated donation and was found to be ELISA‐3 plus RIBA‐3 positive. Eight months after the implicated donation, the donor is still PCR and RIBA‐3 positive, whereas the recipient became PCR negative but remained anti‐HCV RIBA‐3 positive. The case shows that blood products from donors collected during the open window period of an HCV infection can transmit HCV to recipients.
Influence of Haemorrhage and Blood Transfusion on Haemostasis: An Experimental Study in Rabbits
Authors: Henriksson Anders E. , Bergqvist David , Ljungberg June , Hedner Ulla
Published: March 2009

The influence of haemorrhage and blood transfusion on primary haemostasis, coagulation and fibrinolysis was investigated in rabbits. Acute loss of 20% of the blood volume gave a significantly shortened coagulation time (Lee‐White method) but no detectable change in fibrinolysis (euglobulin clot lysis time) and primary haemostasis (primary haemostatic plug formation time in transected arterioles in rabbit mesenteric microcirculation). Acute loss of 20% of blood volume followed by blood transfusion resulted in a prolonged coagulation time and also in a prolonged primary haemostatic plug formation time, but no change in fibrinolysis. It could be concluded that the haemorrhage did not affect the platelet‐dependent primary haemostasis but resulted in a shortened coagulation time. Blood transfusion seemed to affect adversely both the primary haemostasis and the shortened coagulation time induced by haemorrhage.
Molecular Characterization of Glycophorin A Transcripts in Human Erythroid Cells Using RT‐PCR, Allele‐Specific Restriction, and Sequencing
Authors: DuPont B.R. , Grant S.G. , Oto S.H. , Bigbee W.L. , Jensen R.H. , Langlois R.G.
Published: March 2009

Glycophorin A (GPA) is an erythroid‐lineage‐specific membrane sialoglycoprotein which occurs in two allelic forms, M and N, which form the antigens of the MN blood group. Purified cDNAs and RNAs isolated from peripheral blood and erythroleukemia cell lines, HEL and K562, were used to develop an RT‐PCR technique for amplifying GPA gene transcripts (GYPA). The relative expression of transcripts from the M and N alleles was determined using restriction analysis of these amplified products with four allele‐specific restriction endonucleases. The use of this method permits the sensitive identification of GYPA transcripts in these cells and confirms GPA protein expression in the erythroleukemia cell lines and the MN phenotypes of individuals determined by immunolabeling with GPA allele‐specific monoclonal antibodies. A novel restriction pattern was obtained using peripheral blood RNA from two individuals with a rare inherited variant allele, GPA Mg. Sequencing of the cDNA obtained using this method revealed a single C to A transversion in the fourth codon in the mature GYPA N coding sequence is responsible for the difference between GYPA Mg and GYPA N.
Growth Factor Release during Preparation and Storage of Platelet Concentrates
Authors: Ledent E. , Wasteson Å. , Berlin G.
Published: March 2009

The platelet content of platelet‐derived growth factor (PDGF), a mitogen stored in the alpha‐granules, was studied during preparation and storage of platelet concentrates (PC) and compared to the growth‐promoting activity of platelets, β‐thromboglobulin (β‐TG) and lactate dehydrogenase (LD). We compared PC prepared from platelet‐rich plasma (PRP‐PC; n= 10) and from buffy coat. Two different pre‐preparation storage periods of the buffy coat were used: 4h (BC‐PC:4h; n = 10) and 24 h (BC‐PC: 24h; n = 5). The platelet content of PDGF and β‐TG was measured by a RIA technique and the growth‐promoting activity by incorporation of 3H‐thymidine in stimulated fibroblasts. The platelet content of PDGF, β‐TG and the growth‐promoting activity of the platelets decreased in a similar way during preparation and storage of PRP‐PC (31 ±2, 35±2 and 33±7%, respectively, at day 5 of storage; mean ± SEM). The release of LD was minor (3.9 ±0.5% at day 5). At day 1 of storage the platelet content of PDGF was significantly better preserved in BC‐PC:4h than in BD‐PC:24h (88±2 and 81 ±3%, respectively; p = 0.03). Comparing BC‐PC:4h and PRP‐PC we found a significantly better preservation of PDGF in BC‐PC:4h until day 3 of storage (80±2 and 75±1%, respectively at day 3; p = 0.046). In conclusion the preparation of PC according to the PRP method initially induces a higher loss of PDGF, and hence of the growth‐promoting activity, than the BC method.
Comparison of Activity of Peripheral Monocytes and Splenic Macrophages in the Monocyte Monolayer Assay
Authors: Żupańska Barbara , Gronkowska Anna , Ziemski Marek
Published: March 2009

Functional assays intended to reflect the haemolysis in vivo have been performed with peripheral monocytes, occasionally only with peritoneal macrophages or monocyte‐like cell lines. Although the splenic macrophages presumably play the major role in immune red cell destruction, the functional assays have not been done with these cells because of their relative inaccessibility. In this study, the activity of peripheral monocytes and splenic macrophages was compared using the monocyte monolayer assay (MMA), and no substantial differences were found.
Investigation of the Biochemical Relationship between the Blood Group Antigens Xga and CD99 (12E7 Antigen) on Red Cells
Authors: Petty Alison C. , Tippett Patricia
Published: March 2009

The biochemical relationship between the red cell antigens Xgaand the MIC2 gene product, CD99 – previously designated the 12E7 antigen – has been examined by immunoblotting and immunoprecipitation analyses of the protein molecules bearing these antigens. Immunoblotting of membrane components and Xga‐immunoprecipitates with anti‐Xgahas shown that Xgaantigen is carried on a broad band of apparent molecular weight (Mr) 24,500–29,500, which consists of a darkly stained component at Mr 24,500 and a more diffusely stained component at approximately Mr 26,500–29,500. Immunoblotting of membrane components and 12E7‐immunoprecipitates with 12E7, and RFB‐1 and NaM123 which also recognise CD99, distinguished two bands of Mr 30,000 and 32,000. A non‐radioactive immunoprecipitation technique was employed, which uses chemiluminescence detection of biotin‐labelled red cell proteins. The protein of Mr 32,000, which carries CD99, was identified by this method and the red cell quantitative polymorphism of CD99 was demonstrated. When the Xgaprotein was precipitated from biotin‐labelled red cells, a protein of Mr 32,000 was coprecipitated. This suggests that the proteins carrying the Xgaantigen and CD99 are associated in the membrane.
Buffy‐Coat‐Derived Platelet Concentrates Prepared from Half‐Strength Citrate CPD and CPD Whole‐Blood Units: Comparison between Three Additive Solutions: In vitro Studies
Authors: Gulliksson H. , Eriksson L. , Högman C.F. , Payrat J.M.
Published: March 2009

The in vitro effects of storage of platelets prepared from 4 or 6 pooled buffy coat (BC) units and stored in a platelet storage medium consisting of 30–40% of CPD plasma or alternatively half‐strength citrate CPD (0.5 CPD) plasma and 60–70% of different alternative platelet additive solutions (PASs) were evaluated. Measurements of mean platelet volume, pH, pO2, pCO2, bicarbonate, glucose, lactate, ATP, total adenine nucleotide content, extracellular lactate dehydrogenase or adenylate kinase activity, as markers for disintegration of platelets, and extracellular β‐thromboglobulin, as a marker for activation of platelets, were included in the in vitro studies. Previous studies indicated that a reduction of the citrate concentration from the standard 21 to 8 mmol/l is associated with a significant reduction of the consumption of glucose and production of lactate. Alternatively, similar effects can be obtained by the addition of acetate. In a preliminary paired study, the effects of different concentrations of acetate were tested. In an additional paired study, the effects of CPD plasma in combination with either saline or a PAS containing NaCl (115.5 mmol/l), citrate (10 mmol/l), and acetate (30 mmol/l), pH 7.2 (PAS‐2) were evaluated. 0.5CPD plasma in combination with either PAS‐2 or a nonacetate PAS (PAS‐1) were also tested. The storage of platelets in 0.5CPD plasma was used as a reference. The conclusions are: (1) A minimum acetate concentration of 30 mmol/l is needed to counteract the effects of citrate on the production of lactate. (2) pH and the bicarbonate buffering capacity are significantly better maintained in PAS‐2 than in PAS‐1. With PAS‐2, a significantly lower consumption of glucose and a significantly lower production of lactate are also observed. (3) When the storage period exceeds 5 days, PAS‐2 is superior to saline as plasma diluent with regard to the maintenance of pH. (4) A significantly lower consumption of glucose was noted in 0.5CPD plasma diluted with PAS‐2 as compared to undiluted 0.5CPD plasma. A significantly better maintenance of bicarbonate buffering capacity was also seen. (5) By comparing PAS‐2 in combination with either CPD or 0.5CPD, significantly lower values for pCO2 and consumption of glucose were observed with 0.5CPD. The levels of pH and pO2 are significantly higher with 0.5CPD. We conclude that PAS‐2 during 9‐day storage is associated with the most stable storage conditions with regard to the in vitro variables used in this study. The results suggest that at least 7 days storage of PCs in a medium consisting of 30% of either standard CPD or 0.5CPD plasma and 70% PAS‐2 is possible.
Storage of Platelet Concentrates from Overnight‐Stored Blood and Overnight‐Stored Buffy Coat: In vitro Studies
Authors: Rácz Z. , Baróti C.
Published: March 2009

Platelet concentrates (PCs) were prepared from single buffy coats derived from fresh blood and from blood units stored overnight, as well as from buffy coats that were stored overnight. The platelet yield from overnight‐stored buffy coats was similar to that of fresh blood or overnight‐stored blood. PCs were stored at 20–24°C and on day 5 of storage, platelet aggregation with ADP was tested both at 37 and 25°C. Stored platelets aggregated better at 25°C than 37°C. The maximal aggregation (10 μM ADP) of stored platelets from overnight‐stored buffy coats was 46±23% (n = 30), while that of stored platelets prepared either from fresh or overnight‐stored blood was 27±21% (n = 29) and 22±15% (n = 29), respectively. Extracellular lactate dehydrogenase and ammonia levels, as well as elastase activity were similar in stored PCs of different origin. Our conclusion is that PCs prepared from overnight‐stored buffy coat might also be suitable for storage and clinical use. In vivo studies are needed to confirm our findings.
Mixed‐Type Auto‐Immune Haemolytic Anaemia in a Patient with HIV Infection
Authors: De Angelis V. , Biasinutto C. , Pradella P. , Errante D.
Published: March 2009

A young HIV‐infected patient presented with a severe auto‐immune haemolytic anaemia with both warm and cold auto‐antibodies, an infrequent category of anti‐erythrocyte auto‐immunity. Serological findings were compatible with the presence of a low‐titre, high‐thermal‐amplitude anti‐I cold‐reacting antibody and a pan‐reactive warm‐reactive auto‐antibody. Immunochemical characterisation of the warm antibody failed to identify any membrane protein acting as auto‐antigen. This is, to our knowledge, the first reported case of mixed‐type autoimmune haemolytic anaemia in a patient with HIV infection. Overt haemolysis is a very rare complication in HIV‐infected patients, despite the high prevalence of a positive direct antiglobulin test reported in these patients. This suggests that HIV infection is a condition in which anti‐erythrocyte auto‐immunity is a serological finding without haemolytic effects in the large majority of cases.
Bedside Transfusion Errors: Analysis of 2 Years' Use of a System to Monitor and Prevent Transfusion Errors
Authors: Mercuriali F. , Inghilleri G. , Colotti M.T. , Farè M. , Biffi E. , Vinci A. , Podico M. , Scalamogna R.
Published: March 2009

Clerical errors occurring during specimen collection, issue and transfusion of blood are the most common cause of ABO incompatible transfusions. 40–50% of the transfusion fatalities result from errors in properly identifying the patient or the blood components. The frequency and type of errors observed, despite the implementation of measures to prevent them, suggests that errors are inevitable unless major changes in procedures are adopted. A fail‐safe system, which physically prevents the possibility of error, was adopted in January 1993 and concurrently a quality improvement program was implemented to monitor any transfusion errors. Up to December 1994, 10,995 blood units (5,057 autologous and 5,938 allogeneic) were transfused to 3,231 patients. Seventy‐one methodological errors (1/155 units) were observed, half of which were concentrated during the first 4 months of introducing the system. However the system detected and avoided four potentially fatal errors (1/2,748 units). Two cases involved the interchanging of recipient sample tubes, 1 case was due to patient misidentification and the other involved misidentification of blood units. In conclusion the system is effective in detecting otherwise undiscovered errors in transfusion practice and can prevent potential transfusion‐associated fatalities caused by misidentification of blood units or recipients.
Frequent Occurrence of a Variant O1 Gene at the Blood Group ABO Locus
Authors: Olsson Martin L. , Chester M. Alan
Published: March 2009

Blood group ABO polymorphism was analysed in genomic DNA isolated from 150 blood donors by restriction endonuclease digestion of three polymerase chain reaction‐amplified exons in the ABO genes and by sequencing of randomly selected samples. An anomalous O1 allele first described in a cancer cell line is now shown to account for approximately 40% of the O alleles described to date. This is 10 times more frequent than the only other known variant O allele (O2). This variant O1 allele has at least seven point mutations when compared to the consensus gene, in addition to the deletion characterising the normal O1 allele.
PEL, a ‘New’ High‐Frequency Red Cell Surface Antigen
Authors: Daniels G.L. , Simard H. , Goldman M. , Taliano V. , Fleury M. , Decary F. , Spurll G. , Garcia C. , Ouellet P.
Published: March 2009

A ‘new’, inherited, high‐frequency blood group antigen has been named PEL and is numbered 901014. Two PEL‐propositi with anti‐PEL have been found, both French‐Canadians. Two other French‐Canadian propositi with very weak expression of PEL on their red cells have an antibody, provisionally named anti‐MTP, which does not react with PEL‐cells.
Genotype Frequencies of the Human Platelet Antigen, Ca/Tu, in Japanese, Determined by a PCR‐RFLP Method
Authors: Tanaka Shigenori , Taniue Atsuko , Nagao Nobuo , Tomita Tadao , Ohnoki Shiro , Shibata Hirotoshi , Okubo Yasuto , Yamaguchi Hideo , Shibata Yoichi
Published: March 2009

Recently, the polymorphism of a new human platelet antigen, Ca/Tu, was shown to be derived from a G‐A nucleotide substitution at base 1564 of GPIIIa cDNA, which leads to a single amino acid difference, Arg/Gln at amino acid 489 of GPIIIa. We developed a PCR‐RFLP method to determine the genotypes of Ca/Tu and their frequencies in a Japanese population. Fifteen Ca/Tua donors comprising 1 Ca/Tu(a/a) homozygous donor and 14 Ca/Tu(a/b) heterozygous donors were found among the 314 random donors analyzed. The frequencies of Ca/Tu genes were 0.025 (Ca/Tua) and 0.975 (Ca/Tub). The present study showed that the frequency of Ca/Tua individuals in the Japanese (15/314) was approximately 7‐fold higher than in the Finnish population (1/150) previously reported by Kekomäki et al. Therefore, attention must be given to the involvement of the Ca/Tu alloantigen in neonatal alloimmune thrombocytopenia and the refractoriness of platelet transfusion.
Coagulation Factor Activation in Stored Platelet Concentrates is Modulated by the Platelets
Authors: Gyöngyössy‐Issa Maria I. C. , Black Timothy , Devine Dana V.
Published: March 2009

To assess alterations in coagulation proteins in stored platelet concentrates, we used a series of platelet parameters and measures of coagulation activation to compare samples collected before unit donation, during the processing of platelet concentrates (PC) from CPDA‐1 blood, and in storage up to 5 days as well as stored platelet‐poor plasma (PPP). Storage‐dependent increases in activated partial thromboplastin time and prothrombin time were seen in both PC and PPP. However, FVII, FXI, FXII, kallikrein activity and prothrombin F1.2 levels remained unchanged in stored PC. Surprisingly, in stored PPP, significant alterations in FXII, FVII, kallikrein and prothrombin F1.2 levels were seen. Platelet morphology and surface marker studies demonstrated platelet activation during storage. These data suggest that the presence of platelets in the CLX storage container partially suppresses coagulation activation at significant cost to platelet viability.
Donor‐to‐Donor and Donor‐to‐Patient Transmission of Hepatitis C Virus
Authors: Kitchen Alan D. , Wallis Paul A. , Gorman Angela M.
Published: March 2009

A patient was reported with suspected acute post‐transfusion hepatitis C virus (HCV) infection, 5 months after the transfusion of 2 units of red cells. The archived serum samples of the two implicated donations were retested by the original 3rd‐generation assay together with another 3rd‐generation assay and RI‐BA III, and were tested for HCV RNA using the PCR. Both donations were anti‐HCV negative but one was found to be PCR positive. This donor was a regular donor and was identified as the husband of a donor identified 18 months earlier as being HCV positive. This case is an example of transmission of HCV in the window period of infection, and a probable example of the transmission of HCV from wife to husband through intimate contact.
Analysis for Recovery and Loss of Mononuclear Cells and Colony‐Forming Units Granulocyte‐Macrophage during ex vivo Processing of Autologous Bone Marrow
Authors: Schwella N. , Heuft H. G. , Rick O. , Blasczyk R. , Wittmann G. , Huhn D.
Published: March 2009

During ex vivo processing of autologous bone marrow (BM) substantial loss of stem and progenitor cells should be avoided to achieve rapid and sustained hematopoietic reconstitution after high‐dose radio‐/chemotherapy. We processed 25 autologous BM grafts with the Fresenius AS 104 cell separator for cryopreservation and we determined recoveries for mononuclear cells (MNC) and colony‐forming units granulocyte‐macrophage (CFU‐GM) in the BM concentrates. To identify cell loss in BM fractions not cryopreserved, we investigated the MNC and CFU‐GM content of BM fat and BM blood. MNC and CFU‐GM recovery yielded a mean (± SEM) of 42±12 and 54±20% in the BM concentrate. BM fat showed a mean loss of 7±5% for MNC and 4±3% for CFU‐GM, BM blood 30 ±12% for MNC and 13±13% for CFU‐GM, respectively. CFU‐GM recovery was significantly higher in the BM concentrate of patients with hematologic malignancy (HM) compared with patients suffering from germ cell cancer (GCC): 66 ±21 vs. 43±12% (p<0.02). Seventeen patients (7 GCC, 10 HM) underwent high‐dose chemotherapy or radio‐/chemotherapy and were autografted with 0.8 ±0.2times108 MNC/kg and 3.7±2.0×104 CFU‐GM/kg. All patients achieved engraftment with neutrophils >0.5×109/1 at a mean of 14±6 days. We conclude that: (1) ex vivo processing of autologous BM with a mean recovery of 42% for MNC and 54% for CFU‐GM in the BM concentrate can result in a cell population capable of sustained hematopoietic reconstitution, (2) CFU‐GM recovery is significantly higher in patients with HM than in patients with GCC and (3) 37% MNC and 17% CFU‐GM represent in fact cell losses recovered from BM fractions not cryopreserved (BM fat, BM blood). Furthermore, it is likely that MNC and CFU‐GM not recovered from BM concentrate, BM fat and BM blood are cell losses related to the cell separator.
Collaborative Study to Assess the Suitability of an HCV RNA Reference Sample for Detection of an HCV RNA in Plasma Pools by PCR
Authors: Saldanha J. , Minor P.
Published: March 2009

A collaborative study was undertaken to examine the sensitivity and reproducibility of hepatitis C virus (HCV) RNA PCR assays of plasma pools in order to establish a reference sample for HCV PCR testing of plasma pools. Samples consisting of an HCV‐RNA‐positive donation diluted tenfold in an HCV‐RNA‐negative cryosupernatant were sent to 16 participating laboratories (including blood product manufacturers and control authorities). The results of the in‐house assays indicate that a lo‐4 dilution of the positive donation in the cryosupernatant could be used as the reference sample as this was the highest dilution of the positive donation that was detected in all assays. In contrast, the results obtained with a commercially available assay, designed for use with single‐donation plasma or serum (Roche AmplicorTM HCV test), were not so clear‐cut and the assay appeared to be ten‐fold less sensitive than the in‐house assays.
Inhibition of the Monocyte Chemiluminescent Response to Anti‐D‐Sensitized Red Cells by FcγRI‐Blocking Antibodies Which Ameliorate the Severity of Haemolytic Disease of the Newborn
Authors: Shepard S. L. , Noble A. L. , Hadley A. G. , Filbey D.
Published: March 2009

Luminol‐enhanced chemiluminescence (CL) may be used to measure the metabolic response of human monocytes to red cells sensitized by anti‐D. The functional activity of maternal anti‐D when measured in this way correlates with the severity of haemolytic disease of the newborn (HDN) in D‐positive fetuses. Occasionally, however, women with levels of functionally active anti‐D predictive of moderate to severe HDN may deliver D‐positive babies with unexpectedly mild disease. In the current study we have shown that serum from 12 of 15 such women contained monocyte‐binding IgG antibodies which blocked FcγRI and inhibited the CL response of moncytes to red cells sensitized with monoclonal anti‐D. In contrast, FcγRI‐blocking antibodies were found in the serum from only 4 of 11 women who were matched for anti‐D activity but who delivered babies with severe HDN (p < 0.05). Antibodies responsible for inhibition of CL responses were predominantly against HLA class I antigens. The CL response of monocytes to sensitized red cells was at least as sensitive to inhibition by FcγRI‐blocking antibodies as were phagocytic and lytic responses. Our data suggest that inhibition of the CL test is an objective, sensitive and relatively simple technique for detecting and investigating FcγRI‐blocking antibodies which may ameliorate the severity of HDN.
DNA‐Based Rhesus Typing: Simultaneous Determination of RHC and RHD Status Using the Polymerase Chain Reaction
Authors: Poulter Mark , Kemp Tim J. , Carritt Ben
Published: March 2009

We describe a PCR‐based method of performing RHD and C/c typing in a single reaction. The method is based on an earlier observation of a polymorphism in intron 2 of both genes which, in addition to detecting the RHD deletion responsible for most known D‐negative phenotypes, is also associated with C/c serological type. Using this assay, we typed 105 unrelated individuals from at least four different population groups and compared the results to those obtained using conventional serological testing of red cells. An absolute correlation, with no exceptions, was seen. We also showed that the method has potential in the antenatal determination of RH type, as it was possible to type fetal trophoblasts recovered from the endocervical canal at 9 weeks pregnancy.
The D Antigen Characteristic of RoHar Is a Partial D Antigen
Authors: Wallace M. , Lomas‐Francis C. , Tippett P.
Published: March 2009

The results of testing RoHarr cells with panels of monoclonal anti‐D suggest that the D antigen (referred to as DHar) encoded by the haplotype (D)c(e) Rh33 is a partial D antigen. IgM monoclonal anti‐D are more efficient than IgG monoclonal anti‐D in detecting DHar. DHar expresses some but not all of both epD5 and epD6/7 and appears to lack epD1‐epD4, epD8 and epD9.
Current Research on the Immunomodulatory Effect of Allogeneic Blood Transfusion
Authors: Dzik Sunny , Blajchman Morris A. , Blumberg Neil , Kirkley Scott A. , Heal Joanna M. , Wood Kathryn
Published: March 2009

This review summarizes three aspects of current research on the immunomodulatory effect of allogeneic transfusion. Representatives of three laboratories ‐ each of which is actively engaged in research on transfusion‐induced immunomodulation ‐ summarize their current investigative approach. First, current animal models of transfusion‐induced immunomodulation are presented and research on the tumor growth‐promoting effect of allogeneic transfusion is described. Second, mechanisms underlying an immunomodulatory effect of transfusion are summarized and experiments on the induction of transplant tolerance by selective introduction of donor‐type MHC antigens is presented. Third, the potential clinical impact of increased infection and tumor recurrence resulting from transfusion‐induced immunomodulation is assessed. The potential role of donor‐derived hematopoietic stem cells is presented as an area of future investigation in the area of transfusion‐induced immunomodulation.
A Randomised Trial of Fibrin Sealant in Peripheral Vascular Surgery
Authors: Milne A. A. , Murphy W. G. , Reading S.J. , Ruckley C. V.
Published: March 2009

In a prospective randomised trial 39 patients undergoing either arterial bypass surgery with a polytetrafluoroethylene (PTFE) bypass graft (n = 18) or aortic aneurysm repair with a woven Dacron graft (n = 21) were randomised either to receive fibrin sealant as a topical haemostatic agent at the arterial anastomosis or to act as control. The main outcome measure was the time taken to achieve haemostasis at the suture line. The median time to achieve haemostasis was 0.5 min (range 0–11 min) in the treatment group and 4 min (range 0–21 min) in the control group. This difference was statistically significant p < 0.014 by the Mann‐Whitney test. Immediate haemostasis on release of the clamps was achieved in 13/21 patients in the treatment group and in 4/18 patients in the control group (p = 0.023 by Fisher's exact test). There was no difference in total operative time or operative blood loss. No patients in the treatment group suffered any perioperative thromboembolic event and 1 patient in the control group suffered an early graft occlusion. There was no evidence of transmission of hepatitis B or C, or parvovirus B19. In conclusion, fibrin sealant is an effective topical haemostatic agent for arterial suture lines involving PTFE or woven Dacron.
PLASMA‐DERIVED VERSUS RECOMBINANT FACTOR VIII FOR THE TREATMENT OF HEMOPHILIA A
Authors: KASPER CAROL K.
Published: March 2009

Many excellent concentrates now exist for treating hemophilia A. The risk of transmitting donor viral infections with plasma‐derived factor VIII (pdVIII) has been reduced to a very low level; the risk of undetected microbial contamination of recombinant factor VIII (rVIII) probably is much lower, but is not zero. The risk of inducing an excess number of inhibitors (neoantigenicity) may vary among pdVIII brands and may be higher with rVIII than with the least antigenic pdVIII, but the relative risks are unproven. The lowest prices now charged in some areas for excellent pdVIII, or the additional cost in some areas of preparing pdVIII from local blood or plasma donations, are lower than the lowest price charged now or predicted for the future for rVIII. Shortages of both pdVIII and rVIII have occurred. Although pdVIII may remain the mainstay of those countries with limited budgets, and those that want to preserve self‐sufficiency, while rVIII may be preferred by those who can afford its perceived lower risk of infection, multiple options should be kept open.
AN OVERVIEW OF THERAPEUTIC APHERESIS
Authors: BORBERG H.
Published: March 2009

Haemapheresis, or more generally, apheresis is concerned with the separation and differential separation of blood. It differs from dialysis, in that the process of separation concentrates mainly on cells and high molecular weight proteins. Correspondingly different separation techniques are used. Though apheresis can be performed applying conventional off‐line centrifugal techniques, it is generally associated with the use of on‐line centrifugation in blood cell separators (nowadays prefered to be called blood separators), plasmapheresis systems or devices for the preparation of blood components serving autologous blood transfusion. Filtration through flat sheet membranes or hollow fiber membranes is also applied. The differential separation of cells or plasma is off‐line and on‐line, correlating with the different degrees of technical maturation of the secondary separation systems. These secondary techniques aim at a concentration of certain cell types (for instance of stem cells) or at the elimination of undesired cells such as T cells or erythrocytes from stem cell or bone marrow concentrates. In secondary plasma treatement, filtration or adsorption procedures compete with immunological techniques and magnetic binding or density centrifugation or combinations of several techniques.
Genetic Variability of Phytopathogenic Fusarium proliferatum Associated with Crown Rot in Asparagus officinalis
Authors: Bargen Susanne von , Martinez Oliver , Schadock Ines , Eisold Anne‐Mareen , Gossmann Monika , Büttner Carmen
Published: March 2009

Fusarium proliferatum (teleomorph: Gibberella intermedia) is a causal agent of crown rot of Asparagus officinalis and is one potential fumonisin‐producing species within the genus Fusarium. It colonizes roots and crowns of asparagus plants, but could also be isolated from symptomless asparagus spears. Fusarium proliferatum isolates obtained from perennial asparagus plantings from Austria and Germany were included in a study on detectability and variability of two essential genes of the fumonisin‐gene cluster. Genetic fingerprinting of 45 isolates revealed 14 different fingerprint groups, indicating genetic heterogenicity of F. proliferatum. Most isolates differentiated into three main fingerprint clusters, but no association was found between fingerprint group and origin of the isolates. By gene‐specific PCR it was shown that, in 25 isolates tested, both initial genes of the fumonisin biosynthetic pathway –FUM1, encoding a polyketide synthase and FUM8, a gene for a putative aminoacyl transferase – were detectable. This suggests that these isolates were able to produce fumonisins and could contribute to the detected contamination in originating asparagus spears with this mycotoxin. Thus, early detection of FUM‐genes in F. proliferatum‐colonized asparagus may be suited to prevent uptake of fumonisin contaminated food with the human diet. Restriction fragment length polymorphism analysis (PCR‐RFLP) of the amplified FUM gene fragments revealed little sequence variability, suggesting a conserved structure of these genes within this species. However, sequence analysis confirmed intraspecific nucleotide polymorphisms of these genes.
The Grain Mold Pathogen, Fusarium thapsinum, Reduces Caryopsis Formation in Sorghum bicolor
Authors: Little Christopher R. , Magill Clint W.
Published: March 2009

Grain mold is a globally important panicle disease of sorghum [Sorghum bicolor (L.) Moench] that occurs between floral anthesis and physiological maturity of the caryopsis. Many fungi, including Fusarium thapsinum, cause grain mold. In this study, sorghum florets of grain mold resistant (Sureno and Tx2911), moderately resistant (SC170), and susceptible (Tx430) cultivars were spray‐inoculated with a high density (1 × 106 microconidia/ml) of F. thapsinum at anthesis. Mean caryopsis formation frequencies [CFF = (total caryopses/total spikelets per panicle) × 100] for F. thapsinum‐inoculated Sureno and Tx2911 panicles did not significantly differ from their respective controls, however F. thapsinum‐inoculated SC170 and Tx430 panicles were significantly less than controls. In addition, CFF was significantly reduced in F. thapsinum‐inoculated Tx430 panicles when compared to the F. thapsinum‐inoculated grain mold resistant genotypes (P = 0.011). This study suggests that screening of sorghum varieties for caryopsis formation using high‐density inoculations at anthesis would provide an additional assay to measure F. thapsinum resistance in the greenhouse.
Competition Between Cucumber Mosaic Virus Subgroup I and II Isolates in Tobacco
Authors: Tian Zhaofeng , Qiu Jiyan , Yu Jialin , Han Chenggui , Liu Weicheng
Published: March 2009

Historical reports indicate that Cucumber mosaic virus (CMV) strain subgroup I is more prevalent than subgroup II in most parts of the world, but recent reports suggest that subgroup II isolates may be far more abundant than previously found in China. In order to evaluate the dominance of CMV subgroup I and subgroup II strains in co‐infected tobacco plants, four isolates, NX and YQ in subgroup I, and ZL and AG in subgroup II, were tested in competition experiments. In these comparisons, the frequency of infection was assessed, and ratios between singly and doubly infected plants were calculated based on ELISA tests of tobacco leaves. In contrast to previous reports suggesting that subgroup I strains are usually more competitive than subgroup II strains in the field, the results from the present study indicate that the subgroup II ZL isolate was more competitive than the subgroup I YQ isolate, even though the ZL isolate caused milder symptoms than YQ in singly infected tobacco. In contrast, the subgroup I strains NX and YQ were more competitive than subgroup II AG. This information provides evidence for variation in the competitive abilities of subgroup II strains in tests with subgroup I strains, and suggests that direct competition during mixed infections may account in part for the recent spread of some subgroup II strains in China and elsewhere.