bluuebunny/crossref_metadata_2025 huggingface dataset:
Histological and Immunohistochemical Studies on the Preparation of Leukocyte‐Poor Red Cell Concentrates by Filtration: The Filtration Process on Cellulose Acetate Fibers
Authors: Steneker Ingeborg , Biewenga Jeike
Published: March 2009

Abstract. A histological and immunohistochemical investigation of slices from the Cellselect® leukocyte filter showed that the capture of lymphocytes within these filters depended on trapping of the cells in the cellulose acetate fiber network more than on adherence. Probably, additional mechanisms played a role in granulocyte removal. Microaggregates of granulocytes and platelets were found at the top of the Cellselect leukocyte filter. Therefore, it is likely that granulocyte and platelet removal, at least in part, is due to the formation of cell clusters on the surface of the fibers. Although a part of the original, nonfiltered, leukocytes already showed morphologic characteristics of cell necrosis, more necrotic leukocytes were detected within the filter.
Prevalence of Hepatitis C Virus Antibodies in Italian Blood Donors
Authors: Sirchia G. , Almini D. , Bellobuono A. , Giovanetti A. M. , Marconi M. , Mercuriali F. , Mozzi F. , Parravicini A. , Pizzi M. , Zanuso F. ,   The Italian Cooperative Group
Published: March 2009

Abstract. 11,117 blood donors from 24 blood transfusion services evenly distributed throughout the various Italian regions were tested for the presence of hepatitis C virus (HCV) antibodies in the serum and serum alanine aminotransferase (ALT) level. The results are as follows: (1) anti‐HCV seroprevalence in Italy was 0.87% with a difference between Northern and Southern regions (0.68 vs. 1.37%) and between younger and older subjects (0.62 vs. 1.21%); (2) prevalence of elevated ALT levels was 4.74% without a North‐South effect (except than for markedly elevated ALT levels); (3) anti‐HCV seroprevalence was higher in subjects with elevated ALT (5.0%), with a North‐South effect (2.2 vs. 9.9%) and particularly high (19.2%) in subjects with markedly elevated ALT; (4) ALT levels were elevated in 26.2% of anti‐HCV positive subjects, with a North‐South effect (14 vs. 40.5%).
A Low‐Incidence Red Cell Antigen JAL Associated with Two Unusual Rh Gene Complexes
Authors: Lomas C. , Poole J. , Salaru N. , Redman M. , Kirkley K. , Moulds M. , McCreary J. , Nicholson G.S. , Hustinx H. , Green C.
Published: March 2009

Abstract. A multilaboratory investigation during several years has identified a low incidence antigen JAL on the red cells of 7 propositi. JAL appears to be associated with two unusual Rh complexes, one of which produces a depressed C antigen and the other a depressed c antigen. Family studies strongly suggest that the JAL antigen is encoded by the RH locus. Anti‐JAL has been implicated in haemolytic disease of the newborn and is thus considered to be a clinically significant antibody.
Serology and Genetics of the Red Blood Cell Factor Rh34
Authors: Moores Phyllis , Smart Elizabeth
Published: March 2009

Rh34 antibodies were found to have a unique, previously only partly characterised, specificity within the Rh system. No evidence was seen that they were mixtures of hrB and Hr‐like antibodies, or that hrB antibodies existed independently in the natural state. The term anti‐hrB applied to Rh34 antibodies after they had been partially absorbed with R2R2 red cells. Four haplotypes not expressing Rh34 antigen were identified in the present study. The prefix * has been used to indicate them in this text. They were *r's (*dCces), *Ro (*Dce), *Rou (*Duce) and *R od(category III *Dce). Red cells with partially deleted or Rhnull phenotypes were therefore not the sole red cells compatible with anti‐Rh34. R2R2 red cells, which are known to carry weak Rh34 antigen, were incompatible. Twenty‐two family and mother‐child studies established that the Rh:‐34 haplotypes were inherited as normal Mendelian dominant characters. Anti‐Rh34 was capable of recognising RH 34 dosage and of excluding some men who had been wrongly accused in disputed paternity tests.
Antibody to Hepatitis‐C‐Virus‐Related Proteins in Sera from Alanine‐Aminotransferase‐Screened Blood Donors and Prospectively Studied Recipients
Authors: Widell A. , Sundström G. , Hansson B. G. , Moestrup T. , Nordenfelt E.
Published: March 2009

Abstract. A prospective study of posttransfusion non‐A, non‐B hepatitis was conducted in Malmö, Sweden, in 1984–1985, in which donors were alanine aminotransferase (ALT) screened but not ALT selected. Among 741 patients studied at 0, 6, and 12 weeks after transfusion, 13 developed non‐A, non‐B hepatitis, and these were further followed up. Stored sera from the 13 hepatitis patients and their 123 donors were tested for anti‐hepatitis C virus (HCV) by ELISA and, if positive, analyzed by recombinant immunoblot assay (RIBA). All ALT‐elevated blood units (n = 301) and a similar number of ALT‐normal units were also tested. Only 4/13 patients with non‐A, non‐B hepatitis seroconverted to anti‐HCV, all with ALT peaks >10 times the upper normal. All seroconversions occurred within 5 months after transfusion and could be confirmed by RIBA. Hepatitis C in recipients occurred both after transfusion of blood that was strongly positive, weakly positive, and/or negative for anti‐HCV by ELISA. In donors grouped by ALT levels, the anti‐HCV prevalence varied between 0.4 (normal ALT) and 14% (ALT elevated ≥ 2 times). Of the total of 9 donor units positive by ELISA, only 5 were confirmed by RIBA. Of the 5 recipients of the RIBA‐positive blood units, 3 went into hepatitis, 1 remained normal at 10.5 weeks, and 1 showed a slight, transient ALT elevation at week 12. The recipients of ELISA‐positive but RIBA‐negative blood remained healthy.
Photoinactivation of Viruses in Human Fresh Plasma by Phenothiazine Dyes in Combination with Visible Light
Authors: Lambrecht Bernd , Mohr Harold , Knüver‐Hopf Josef , Schmitt Heinz
Published: March 2009

Abstract. We developed a photodynamic method to inactivate viruses in human fresh plasma. Single plasma bags were illuminated with visible light in the presence of low doses of phenothiazine dyes like methylene blue or toluidine blue. By this treatment the infectivity of different enveloped viruses including the causative agent of AIDS, HIV‐1, was completely removable from the plasma. Non enveloped viruses, however, proved to be more stable. The activities of clotting factors and other plasma proteins were only slightly decreased. There was no indication that the procedure led to important structural modifications of plasma proteins. The dyes are photodynamically active at concentrations much lower than those at which they are therapeutically used as antidots in the treatment of methemoglobinemia.
Brb, a Platelet Alloantigen Involved in Neonatal Alloimmune Thrombocytopenia
Authors: Bettaieb Ali , Fromont Patricia , Rodet Martine , Godeau Bertrand , Duedari Najib , Bierling Philippe
Published: March 2009

Abstract. Serum from a pregnant woman with the May‐Hegglin anomaly contained a platelet‐specific antibody. The serum reacted in the platelet indirect immunofluorescence test (PIIFT) with 97.6% of random donor platelets and those of the father but not with the mother's own platelets. This antibody induced a moderate thrombocytopenia in the infant that responded to infusion of intravenous immunoglobulin concentrates. The platelet phenotypes were PLA1+, Baka+, Bra+/Brb‐ for the mother, PLA1+, Baka+, Bra‐/Brb+ for the father, and PLA1+, Bra+/Brb+ for the neonate. Analysis of the maternal serum with an immunoassay based on monoclonal antibody immobilization of platelet antigens (MAIPA) and immunoprecipitation techniques demonstrated the absence of antibodies directed against HLA class I antigens and that the antigen recognized was located on the platelet‐GpIa/IIa complex. This antigen was present on 113/115 random donor platelets, in 7 of the 7 unrelated May‐Hegglin platelets, and only absent in 3/24 Bra+ individuals, including the mother. No platelet‐specific antibody was present in the serum of the 7 unrelated May‐Hegglin subjects. The antigen recognized by this platelet‐specific antibody thus meets the criteria defining the antithetic allele of Bra, i.e. the Brb alloantigen.
Studies in Red Blood Cell Preservation: 4. Plasma Vesicle Hemoglobin Exceeds Free Hemoglobin
Authors: Greenwalt T.J. , McGuinness C.G. , Dumaswala U.J.
Published: March 2009

Abstract. Studies were designed to find out how much of the plasma hemoglobin (Hb) in whole blood was in microvesicles and how much was free Hb after 21 days of storage in citrate‐phosphate‐dextrose anticoagulant and to determine the effect of the plasticizer, di‐(2‐ethylhexyl)phthalate (DEHP). The total plasma Hb in polyolefin (PO) containers without DEHP was much higher than in polyvinyl chloride (PVC) with the plasticizer (p=0.004). Less than 30% of the Hb was in free solution in either type of container. The addition of 300 μg/ml of DEHP to the plasma in the PO containers resulted in marked reduction in the microvesiculation (p<0.01) but did not affect the level of free Hb. RBC hypotonic fragility and morphology scores were significantly improved. It is concluded that microvesiculation contributes more to plasma Hb concentration than free Hb during storage.

Some hemolysis of red blood cells (RBC) is expected during blood bank storage. It has been shown that part of the hemoglobin (Hb) in the suspending medium is free and part is encapsulated in microvescicles shed by the RBC [1]. The amount of hemolysis and microvesiculation that occurs has been noted to be less when blood is stored in polyvinyl chloride (PVC) containers in which di‐(2‐ethylhexyl)phthalate (DEHP) is used as the plasticizer [1]. The DEHP that leaches into the plasma has been shown to decrease hemolysis, microvesiculation, and the increase in osmotic fragility which RBC undergo during refrigerated storage [2–9]. The posttransfusion survival of RBC stored in DEHP‐plasticized PVC has been found superior to RBC stored in PVC bags plasticized with tri‐2‐ethylhexyl‐trimellitate (TEHTM) which is much less leachable. The protective effect of the DEHP is believed to be due to its insertion into the RBC membrane [6–9].

The present studies were done to quantify how much of the ‘hemolysis’ that has been observed during the storage of RBC is dissolved Hb and how much is Hb encapsulated in microvesicles.
Immunochemical Characterisation of the Low‐Incidence Antigen, Dha
Authors: Spring F.A.
Published: March 2009

Abstract. Immunoblotting with two examples of anti‐Dha to the electrophoretically separated components of antigen‐positive membranes gave a positive reaction with a component of the same apparent Mr (40,000) as sialoglycoprotein β (SGP β, syn: glycoconnectin, glycophorin C). The Dha antigenic determinant was sensitive to trypsin, but resistant to chymotrypsin and Endo F. By immunoblotting, one anti‐Dha failed to react with sialidase‐treated Dh(a+) cells, whilst the other gave a positive result. In contrast, neither antibody agglutinated sialidase‐treated red cells. SGP β was precipitated from Dh(a+) and Dh(a‐) phenotype red cells by monoclonal anti‐β (NBTS/BRIC 10). SGP β from Dh(a+) but not from Dh(a‐) red cells was stained by immunoblotting with anti‐Dha. These results assign the Dha antigenic epitope to SGP β.
Storage of Platelets in a New Plastic Container: Polyvinyl Chloride Plasticized with Butyryl‐n‐Trihexyl Citrate
Authors: Gulliksson H. , Shanwell A. , Wikman A. , Reppucci A.J. , Sallander S. , Udén A.M.
Published: March 2009

Abstract. The effect of storage of platelets in a new polyvinyl chloride (PVC) plastic material with a butyryl‐n‐trihexyl citrate (BTHC) plasticizer (PL 2209) was evaluated. The PL 1240 container, i.e. PVC plastic with a different plasticizer, tri‐(ethylhexyl)‐tri‐mellitate, was used as a reference. Measurements of pH, pO2, pCO2, glucose, lactate, adenosine triphosphate, total adenine nucleotide content, lactate dehydrogenase and platelet factor 4 (PF4) were made during 5 days of storage. Similar results were noted comparing PL 2209 and PL 1240. Differences in pO2 and pCO2 indicate greater gas permeability in PL 2209 than in PL 1240. Significantly higher PF4 levels were found in PL 2209, but the difference could not be attributed to the PL 2209 container itself. Paired autologous reinfusion studies (111Indium) of 6 normal donors gave mean recovery values after 5‐day storage of 41.1 ± 7.4% (PL 2209) and 45.5 ± 7.7% (PL 1240), t1/2 66 ± 13 and 75 ± 5 h, survival time (linear model) 6.3 ± 1.0 and 6.8 ± 0.7 and survival time (multiple‐hit model) 6.0 ± 0.7 and 6.5 ± 0.4 days, respectively. Only the difference in survival time (multiple‐hit) was significantly higher in PL 1240. The corrected count increments at 12–24 h following transfusion were 13,300 ± 10,800 (PL 2209) and 13,600 ± 11,600 (PL 1240) with no statistically significant difference found. These results indicate PL 2209 as an equivalent alternative to PL 1240 for the 5‐day storage of platelets.
Characterization and Quantification of Anti‐T in Human Serum Using a Reliable Hemolysis Test
Authors: Lynen R. , Simson G. , Rumpf K. W. , Neumeyer H.
Published: March 2009

Abstract. The need of fresh‐frozen donor plasma with a low level of anti‐T has been emphasized recently. Anti‐T, as administered by transfusion of fresh‐frozen plasma, has been accused repeatedly of enhancing hemolysis in septic children with T transformation of red cells. Therefore, a new hemolysis test for the quantification of anti‐T in human serum has been developed. With our test, anti‐T‐poor plasma donors can be found. Additional results raise substantial doubt as to the pathogenetic role of anti‐T in the development of the hemolytic‐uremic syndrome, found in septic children with red‐cell T transformation. It is impossible to predict in vivo hemolysis induced by anti‐T knowing the temperature characteristics and the ionic conditions causing this antibody to mediate hemolysis in vitro. Obviously, T transformation itself plays the major pathogenetic role in these patients, and not the presence of anti‐T. In the case of disseminated intravascular coagulation, a content of anti‐T cannot be construed as prohibiting transfusion of fresh‐frozen plasma to such patients.
Isoagglutinins following ABO‐Incompatible Bone Marrow Transplantation
Authors: Wernet Dorothee , Mayer Gerhard
Published: March 2009

Isoagglutinins were investigated following ABO‐incompatible bone marrow transplantation. In major incompatibility anti‐A tends to require more time to disappear than anti‐B. Correspondingly, A erythrocytes require more time to become demonstrable. There is no such difference in major plus minor incompatibility. In minor incompatibility, isoagglutinins against the recipient blood group may be produced early after bone marrow transplantation, which then give way to a possibly lifelong tolerance for the recipient's old blood group.
Expression of Red Cell Antigens C and c on Neonatal and Adult Erythrocytes: A Flow‐Cytometric Analysis
Authors: DePalma L.
Published: March 2009

Red cell membrane proteins expressing Rh antigens appear to be important for the stabilization of the phospholipid membrane bilayer. Neonatal red cells possess a shorter in vivo life span and different deformability properties as compared to adult red blood cells, differences which could potentially be explained by different cell surface antigen densities of Rh‐antigenic determinants. Although conventional serologic techniques have demonstrated Rh antigens on fetal and neonatal red cells, other more sensitive techniques are required when quantitative information is needed. Therefore, in this study, 40 adult and 45 neonatal red blood cell suspensions of differing Rh phenotypes were evaluated for C and c antigen density using flow cytometry.

Neonatal and adult red blood cells had the same mean channel fluorescence intensity (expressed in mV) of c antigen in either the homozygous (cc) or heterozygous (cC) state: 485+44 versus 471+29 and 416+31 versus 411+23, respectively. In addition, neonatal and adult C antigen density was similar in heterozygous (Cc; 415+49 versus 455+44) and homozygous (CC; 438+53 versus 469+38 C states. As expected, homozygous (double dose) states for either C or c had greater intensities than heterozygous states for both neonates and adults.

Although no differences were noted for C or c antigens, studies of other Rh antigens may indicate lower antigenic density in neonatal red cells.
Diversity of Human Anti‐D Monoclonal Antibodies Revealed by Reactions with Chimpanzee Red Blood Cells
Authors: Blancher A. , Socha W. W. , Ruffie J.
Published: March 2009

Fifty‐three human anti‐D monoclonal antibodies (mAbs) revealed a striking diversity of reactions in tests with panels of chimpanzee red blood cells (RBCs) of various R‐C‐E‐F blood group phenotypes (counterparts of the human Rh‐Hr groups). The reactivities of these antibodies, which depended on the agglutination technique used, could be classified into four main types. These patterns of reactivity of anti‐D mAbs with chimpanzee RBCs showed only limited correlation with types of reactions observed with human D variant RBCs. Primate red cells may, therefore, constitute an independent test system for subclassification of human monoclonal antibodies. Comparison of reactivities of human anti‐D mAbs with chimpanzee and human D variant RBCs confirms the homology between the chimpanzee Rc, and the human D antigens. The chimpanzee Rc shares with human D the epitopes epD5, epD6/7 and epD8, but lacks epitopes epD1, epD2, epD3 and epD4 of the Rh mosaic, thus resembling the human D variants IVb and Vc.
Stored Blood Components Contain Agents That Prime the Neutrophil NADPH Oxidase through the Platelet‐Activating‐Factor Receptor
Authors: Silliman Christopher C. , Thurman Gail W. , Ambruso Daniel R.
Published: March 2009

Neutrophils (PMNs) initiate production of toxic oxygen metabolites through stimulation of an NADPH oxidase resulting in the reduction of oxygen to superoxide anion (O2−). The activity of this enzyme system may be primed by a variety of compounds. This laboratory investigated the possibility that stored blood components may contain agents which prime the PMN oxidase. At the time of outdate, packed red blood cells, whole blood, and platelet concentrates contained a priming agent which enhanced the PMN NADPH oxidase activity in response to a soluble stimulus, formyl‐Met‐Leu‐Phe, by 2.1‐to 2.8‐fold. The priming activity was almost completely inhibited by WEB 2170, a specific platelet activating factor antagonist. Fresh plasma or fresh‐frozen plasma did not exhibit priming activity. These data suggest that platelet‐activating factor‐like compounds are generated during the storage of cellular blood components. The presence of these agents in stored blood may suggest a role for specific componds which prime PMNs and possibly mediate other effects which result in severe complications of transfusion therapy.
Manufacture and in vitro Characterization of a Solvent/Detergent‐Treated Human Plasma
Authors: Hellstern Peter , Sachse Hans , Schwinn Horst , Oberfrank Klaus
Published: March 2009

We have developed a modified solvent/detergent (S/D) treatment to inactivate viruses in human plasma using 1% w/w final concentrations of tri(n‐butyl) phosphate (TNBP) and Triton X‐100 and an incubation period of 4 h at 30°C. The procedure inactivates ≥106 chimpanzee‐infectious doses (CID50) of HBV, ≥105 CID50 of HCV, and ≥106.2 tissue culture infectious doses (TCID50) of HIV. After virus inactivation, eleven plasma batches were lyophilized and 12 batches were deep‐frozen until further use. The batches were characterized by extensive laboratory tests including measurement of clotting factors I–XIII, von Willebrand factor, plasminogen, inhibitors of blood coagulation and fibrinolysis, and other clinically important plasma proteins. All parameters were determined before and after S/D treatment. Twelve conventional single donor plasma units served as control. There were no marked losses of activities of clotting factors, antithrombin III, protein C, plasminogen, and C1‐esterase inhibitor due to treatment. After the S/D step, the levels of these parameters were within the normal range in all batches. The same holds true for total protein, immunoglobulins, albumin, complement factors C3 and C4, haptoglobin, hemopexin, caeruloplasmin, α1‐antitrypsin, and pH. Protein S and α2‐antiplasmin activites decreased by about 50% and were frequently found to be slightly below the lower limit of the respective normal range after treatment. The interindividual variations of all proteins analysed were significantly lower than in the single donor plasma units. The S/D procedure did not lead to increases of markers indicating activation of hemostasis. We conclude that lipid‐enveloped viruses can be inactivated by the S/D procedure described in this study without critical reduction of recoveries of plasma proteins.
Immediate Haemolytic Transfusion Reaction Due to Anti‐Inb
Authors: Joshi S.R.
Published: March 2009

An immediate haemolytic transfusion reaction was investigated in a patient with intestinal cancer. The causative antibody was directed against a high‐frequency antigen Inb that was presumably produced as a result of pregnancies. This is the first case of a severe transfusion reaction due to this allo‐antibody.
Prevalence of Platelet Transfusion Reactions Before and After Implementation of Leukocyte‐Depleted Platelet Concentrates by Filtration
Authors: Goodnough Lawrence Tim , Riddell James , Lazarus Hillard , Chafel Theresa L. , Prince Greg , Hendrix Donna , Yomtovian Roslyn
Published: March 2009

To determine the impact of platelet leukodepletion by filtration on the overall prevalence of reported transfusion reactions associated with platelet concentrates, we audited platelet transfusion reactions after infusion of platelet concentrates reported at University Hospitals of Cleveland over 6 months before (interval 1, July 1, 1989 to December 31, 1989) and after (interval 2, July 1, 1990 to December 31, 1990) implementation of the Pall PL 50 filter on our adult Hematology‐Oncology inpatient unit (Division 60). Thirty‐two (1.7%) of 1,901 random, pooled platelet transfusion events resulted in blood bank transfusion reaction workups in interval 1, compared to 90 (5.3%) of 1,704 in interval 2 (p<0.001). The Division 60 service accounted for more of our hospital‐wide platelet reactions after implementation of the filter in interval 2 (84%) than before filtration in interval 1 (42%), p = 0.002. The prevalence of reaction workups for Division 60 was 0.6% for interval 1, compared to 4.3% for interval 2 (p<0.001). No differences were found between interval 1 and interval 2 for the rate of discontinuation of platelet transfusion (36 vs. 32%, p = 0.14), rate of premedication for platelet transfusion (72 vs. 65%, p = 0.6), percentage of direct antiglobulin test‐positive reactions (17 vs. 5.4%, p = 0.09), percentage showing icteric/hemolyzed serum (15 vs. 4.4%, p = 0.09), or reactions believed to be due to red blood cell incompatibility (8.8 vs. 1.1%, p = 0.1). We conclude that the use of expensive platelet filtration devices has not decreased the morbidity of random, pooled platelet transfusions, nor the prevalence of time‐consuming blood bank evaluation of platelet transfusion reactions in this setting.
Use of Jacalin as a Solid Phase in ABO Reverse Grouping
Authors: Green F. , Bubb M. O. , Conradie J. D.
Published: March 2009

A major problem of using red cells as the solid phase in assay systems is the difficulty to bind them strongly to appropriate surfaces. We report here on a number of lectins of different specificities which were examined for their ability to bind red cells to polystyrene 96‐well microtitre plates. The use of the Thomsen‐Freidenreich antigen‐specific lectins, jacalin, mushroom and Madura pomifera agglutinin proved the most useful for ABO reverse grouping. Jacalin‐coated plates were also compared with plates coated with poly‐L‐lysine and bovine serum albumin/glutaraldehyde for the binding of erythrocyte membranes and were found to be superior. We also describe the col‐orimetric detection of the solid phase red cell antibody reaction by using an indicator erythrocyte and peroxidase chromogenic substrate.
Carbamazepine (Tegretol)‐Induced Thrombocytopenia
Authors: Shechter V. , Brenner B. , Klein E. , Tatarsky I.
Published: March 2009

A case of carbamazepine‐induced thrombocytopenia is reported. The patient, a 38‐year‐old female, developed thrombocytopenia 10 days after receiving 400 mg carbamazepine daily. The presence of an anti‐IgG carbamazepine‐dependent platelet‐reactive antibody was documented. Using flow cytometry, the drug‐induced antibody was detected at a drug concentration as low as 10‐6 M.
Molecular Genetic Analysis of the ABO Blood Group System: 2. cis‐AB Alleles
Authors: Yamamoto Fumi‐ichiro , McNeill Patricia D. , Kominato Yoshihiko , Yamamoto Miyako , Hakomori Sen‐itiroh , Ishimoto Seiji , Nishida Sachiyo , Shima Masayuki , Fujimura Yoshihiro
Published: March 2009

We have determined the nucleotide sequence of the coding region in the last two coding exons of ABO genes from two cis‐AB individuals (genotype cis‐AB/O) with no consanguinity. In this region, cis‐AB alleles from these 2 individuals were identical to one another while different from the A1 allele by two nucleotide substitutions. Both of these nucleotide substitutions result in amino acid substitutions. The first substitution is identical to the one previously found in the A2 allele. The other substitution is found at the fourth position of the four amino acid substitutions which discriminate A1 and B transferases.
Molecular Mimicry between HIV‐1 and Antigen Receptor Molecules: A Clue to the Pathogenesis of AIDS
Authors: Süsal Caner , Kröpelin Marianne , Daniel Volker , Opelz Gerhard
Published: March 2009

There is increasing evidence that autoimmune phenomena play an important role in the immunopathogenesis of AIDS. We found a high degree of sequence homology between HIV‐1 and antigen receptor molecules, immunoglobulins and T cell receptors. Based on recent findings that the appearance of anti‐Fab autoantibodies and attachment of gp120/immunoglobulin/complement complexes on CD4+ T cells are associated with the decrease of CD4+ T cells in HIV‐infected patients, we hypothesize herein that cross‐reactive anti‐F (ab′)2 autoantibodies and circulating gp120 molecules are responsible for a destabilization of the immune network and the elimination of CD4+ T cells.
Hepatitis C Virus Confirmation in Blood Donor Screening
Authors: Waumans L. , Claeys H. , Verhaert H. , Mertens W. , Vermylen C.
Published: March 2009

A combination of different enzyme immunoassays (EIAs) was used for the serological confirmation of sera that were positive in a hepatitis C virus (HCV) second‐generation screening EIA. Different reaction patterns were related with the probability of the HCV‐carrier state as determined by polymerase chain reaction (PCR). Five hundred and eight sera of volunteer blood donors were send for confirmation and at first reexamined with both Abbott and Ortho second‐generation screening EIA. A group of 195 sera, positive in both assays, was further evaluated by the Abbott Supplemental Assay, the Monolisa anti‐HCV and an EIA with only the amino terminal part of the nucleocapsid protein as antigen. In addition PCR on the 5′‐noncoding region of the viral genome was performed. We observed that 75 of the 78 PCR‐positive sera were found in a group of 89 sera that were strongly positive in the four EIAs used. Moreover all but 1 PCR‐positive sera were reactive against the nucleocapsid protein of the virus. Hence we concluded that a genuine antibody response to the nucleocapsid protein is highly suggestive for the HCV‐carrier state.
Evidence for Expression of the Joa Blood Group Antigen on the Gya/Hy‐Active Glycoprotein
Authors: PhD Frances A. Spring , Reid Marion E. , Nicholson Gordon
Published: March 2009

The phenotypic association between the non‐assigned high‐incidence antigen Joa and the Gya collection antigens Gya and Hy was investigated by haemagglutination studies, flow cytometric analysis, immune precipitation and immunoblotting experiments. In haemagglutination tests anti‐Joa gave the same pattern of reactivity with erythrocytes pre‐treated with pronase, trypsin, α‐chymotrypsin and thiol reducing agents as did anti‐Gya and anti‐Hy. In addition, similar to that found for anti‐Gya and anti‐Hy, anti Joa also showed reduced binding, as determined by haemagglutination and flow cytometric analysis, to erythrocytes from patients with paroxysmal nocturnal haemoglobinuria. Immune precipitates prepared from radio‐iodinated antigen‐positive red cells with anti‐Joa, anti‐Gya and anti‐Hy gave similar results – a major component of Mr 49,000–60,000 (the Gya/Hy‐active glycoprotein) and a second component of Mr 85,000–92,000 (this may be a dimer of the Gya/Hy‐active glycoprotein, or a coprecipitated protein). These immune precipitates, when probed with both anti‐Gya and anti‐Hy under non‐reducing conditions, gave a positive immunoblotting reaction to both the Mr 49,000–60,000 and the Mr 85,000–92,000 components. These results strongly suggest that the Joa antigen is expressed on the same glycoprotein that carries the Gya and Hy antigens.
Development and Small‐Scale Production of a Severely Heated Factor VIII Concentrate
Authors: Knevelman A. , De Wit H.J.C. , Potstra P. , Does J.A. v.d.
Published: March 2009

The small‐scale production of a severely heated factor VIII concentrate is described. As starting material for the production 48 units of fresh frozen plasma from whole‐blood donations or 24 units of plasma from apheresis are used. During a glycine and sodium chloride fractionation, contaminating proteins are removed from a pooled extract of single‐donor cryoprecipitates. The resulting factor VIII‐rich precipitate is redissolved in freeze‐drying buffer and this solution is desalted by diafiltration. After filtration, this solution is dispensed into 10 vials and frozen. The product is freeze dried, and heat treated at 80°C for 72 h. The mean yield of the heat‐treated product improved from 160 IU factor VIII per liter plasma at the start of the production to 215 IU per liter plasma after a modification of the purification process. The validation of the virus inactivation during freeze‐drying and 80°C heat treatment for 72 h showed a reduction of ≥7.6 log PFU/ml for Sindbis and a reduction of ≥6.4 log TCID50/ml for HIV‐1. The factor VIII concentrate was clinically tested in 6 patients. It possessed a normal half‐life, showed a high recovery and no side effects. A Dutch license was granted in June 1991 and since then this product has been routinely manufactured in three Dutch blood banks.
A Simple Technique for Red Blood Cell Removal in Major ABO‐Incompatible Bone Marrow Transplantation
Authors: Mayer Gerhard , Wernet Dorothee , Northoff Hinnak , Schneider Werther
Published: March 2009

A simple technique for red blood cell (RBC) removal in major ABO‐incompatible bone marrow transplantation is reported requiring two centrifugation steps, special blood bags and a mechanical device to separate the buffy coat from RBCs within the bag. In 42 transplantations an average of 84% of nucleated cells was recovered with an average contamination of 7.5 ml packed RBCs. The preparations were well tolerated in all patients whose isoagglutinin titers had not been reduced. Bone marrow engraftment was not significantly different from control groups.
Autologous Blood Donation Elective Surgery in Children
Authors: Tasaki Tetsunori , Ohto Hitoshi , Noguchi Mayumi , Abe Rikiya , Kikuchi Shinichi , Hoshino Shunichi
Published: March 2009

Studies were made on 59 children (cardiac 42, orthopaedic 13, miscellaneous 4) scheduled for autologous blood donation before elective surgery. The donor‐patients' ages ranged from 3 to 15 years (mean 9.9 years) and their weights from 13 to 70 kg (mean 34 kg). All patients received 50–100 mg of oral iron sulphate per day. As a rule, about 10% of intravascular blood volume was drawn once a week. Before surgery, an average of 720 ml of autologous blood per patient was prepared. Two patients failed to donate autologous blood because of anxiety about the procedure; however, none of the donors was deferred due anaemia assoicated with the phlebotomy. Of the 53 patients undergoing surgery and participating in autologous predonation, 50 (94%) were able to avoid homologous blood transfusion. 600 ml of homologous blood were transfused to each of 2 orthopaedic patients and 400 ml to 1 cardiac patient. We conclude that a predeposit autologous transfusion programme is logistically possible in small children when the patients are cooperative.
Use of ‘Split’ Plateletpheresis Products for Alloimmunized Patients
Authors: Gharpure Vishwanath S. , Norris Dottie , Lee Edward J. , Schiffer Charles A.
Published: March 2009

Single donor platelets obtained using the COBE Spectra were split into two halves and administered to alloimmunized patients with leukemia at two different points in time. The mean platelet yield was 6.83 times 1011(n = 63) with 56% of collections having yields of >6.0times1011(i.e. twice the current AABB recommendation for an apheresis platelet transfusion). In 58 instances, the two halves were administered to the same patient 24–96 h apart. The differences in the correct increments of the split transfusions administered 24–48 h apart were not statistically significant, although there were decreased increments after 72 and 96 h of storage. In 5 instances when the CCI for first transfusion was unacceptably low, the second half was successfully transfused to another patient. This study proves it is feasible to split apheresis platelets into two transfusions, and discusses approaches to optimally use this strategy for the transfusion support of alloimmunized patients.
Detection and Characterization of Lewis Antigens in Plasma of Lewis‐Negative Individuals Evidence of Chain Extension as a Result of Reduced Fucosyltransferase Competition
Authors: Henry S.M. , Oriol R. , Samuelsson B.E.
Published: March 2009

Nonacid plasma glycolipids from Lewis‐negative individuals of nonsecretor, partial‐secretor and secretor phenotypes were prepared and separated by thin‐layer chromatography and immunostained with radiolabelled Lewis antibodies. Lewis‐positive plasma and intestinal epithelial cell glycolipids from Caucasians representing the four recognized Lewis and secretor combined phenotypes were used as controls. By presenting these purified total glycolipids in a cell‐free environment to Lewis antibodies we were able to demonstrate the presence of small amounts of Lewis antigens in Lewis‐negative individuals. It is shown that lactotetraosylceramide and extended precursor glycolipids are present in all Le(a–b–) nonsecretors. Leawas detected in 1 of the 3 Le(a–b–) nonsecretor plasmas and in the intestinal sample of the same phenotype. Lactotetraosylceramide was absent but H type 1 and Lebwere both present in all group O Le(a–b–) secretors, and extended H type 1 reactive structures were also found in the partial secretor. These results clearly demonstrate that although the Lewis‐negative phenotype exists at the serological level, this phenotype is not an ‘all‐or‐nothing’ phenomenon at the chemical level. We also show that in the presence of reduced fucosyltransferase activity, increased elongation of the precursor chain occurs, which allows us to postulate that fucosylation of the precursor prevents or at least markedly reduces chain elongation.
Prevalence of Infection with the Hepatitis C Virus among Italian Hemophiliacs before and after the Introduction of Virally Inactivated Clotting Factor Concentrates: A Retrospective Evaluation
Authors: Morfini M. , Mannucci P.M. , Ciavarella N. , Schiavoni M. , Gringeri A. , Rafanelli D. , DiBona E. , Chistolini A. , Tagliaferri A. , Rodorigo G. , Baudo F. , Gamba G.
Published: March 2009

In July 1985, all coagulation factor concentrates were withdrawn from the market in Italy and replaced with virally inactivated concentrates. A retrospective survey comparing the prevalence of the antibody to the hepatitis C virus (anti‐HCV) in hemophiliacs multitransfused with nonvirally inactivated concentrates until 1985 with that in previously untreated hemophiliacs transfused exclusively with virally inactivated concentrates since 1985 has been conducted in 9 Italian hemophilia centers. The centers, which follow about one‐fourth of all the Italian hemophiliacs, provided information about 708 patients infused for the first time before 1985 (group A) and 80 patients infused for the first time between 1985 and 1991 (group B). The prevalence of anti‐HCV was 83% (591/708) in group A and 6% (5/80) in group B. For the 5 anti‐HCV‐seropositive patients from group B, dry heating, hydrophobic interaction chromatography plus dry heating (2 patients), hot vapor and pasteurization were the virucidal methods used for the concentrates implicated in HCV transmission. In the case associated with pasteurization, there is the possibility of intrafamilial transmission of HCV. It appears from this retrospective analysis that there has been a substantial reduction in the risk of HCV transmission since the adoption of virucidal methods. However, these methods do not eliminate completely the risk, which might be further reduced by the recent adoption of anti‐HCV screening for plasma donations used to manufacture concentrates.
High‐Dose Intravenous Immunoglobulin in Non‐ABO Transfusion Incompatibility
Authors: Kohan A.I. , Niborski R.C. , Rey J.A. , Amerise G. , Vázquez M.I. , Zani N. , Calahonra R. , Binaghi A. , Avalos J. Sánchez
Published: March 2009

The administration of intravenous immunoglobulin (IVIG) in immune and autoimmune diseases led us to use this agent to ameliorate or prevent the consequences of non‐ABO incompatible transfusions in patients who need this form of therapy. IVIG (400 mg/kg/day) was infused within 24 h of transfusion in 5 patients with: (1) intestinal angiodysplasia, gastrointestinal bleeding, and anti‐Kpb; (2) paroxysmal nocturnal hemoglobinuria, anti‐c, anti E, Anti Fyb, anti‐K and autoantibodies; (3) lymphoma and autoimmune hemolytic anemia (AIHA); (4) systemic lupus erythematosus (SLE), AIHA, and anti‐D, and (5) SLE and AHIA. A sustained increase in hematocrit was noted and no trasfusion reaction developed in any of the cases. A single dose of pretransfusion IVIG may therefore be a useful therapeutic alternative in patients for whom no compatible blood is available. Patients with severe anemia, allo‐ and autoantibodies, either showing hemolysis in their pathophysiology or not, cause a serious problem in any transfusion center, especially when dealing with emergencies. In order to reduce the risks of incompatible transfusions, different modalities have previously been attempted, all with poor results. In 1989 we reported the successful use of pretrans‐fusional high‐dose intravenous immunoglobulin (IVIG) in a patient with gastrointestinal bleeding and anti‐Kpb [19]. The transfusion of incompatible red blood cells improved the anemia and allowed the exploratory laparotomy to take place. A protocol was then developed based on this case administering pretransfusion IVIG in high doses for patients for whom no compatible blood (non‐ABO) is available.
Evaluation of Histo‐Blood Group ABO Genotyping in a Danish Population: Frequency of a Novel O Allele Defined as O2
Authors: Grunnet Niels , Steffensen Rudi , Bennett Eric Paul , Clausen Henrik
Published: March 2009

Traditional blood group ABO serology is based on immunoreactivity with the carbohydrate determinants A, B and H antigens. Recent advances at the DNA level of the ABO genes have provided a molecular genetic model for the ABO polymorphism. This genetic model has to date only been tested on a limited basis. The present study was initiated to evaluate the universality of the proposed genetic model on a larger group of serologically defined ABO phenotypes. Three hundred healthy Danish blood donors were analysed (A: 50, B: 50, AB: 50, O: 150) by PCR amplification followed by diagnostic restriction enzyme cutting. In all cases A, B, and AB at least one allele of correctly predicted status was found. However, in O phenotype individuals, 11 out of 150 carried one allele discordant to the proposed genetic model. This novel O allele (3.7% allele frequency) was further characterized by diagnostic restriction enzyme analysis in two positions divergent between A and B alleles and by DNA sequencing of the two major exons. The novel O allele is termed O2 as it typed as B in nucleotide position 526 and as A in positions 703, 796, and 803, in contrast to the most predominant O allele termed O1, which types as A in all 4 positions. The structural defect in the O2 allele appears to be an additional substitution at nucleotide position 802. The results clearly demonstrate that with the addition of the two distinctly different O alleles, O1, O2, the previously proposed molecular genetic basis of the ABO polymorphism is quite valid. More importantly the determined characteristics of these two O alleles have practical implications in ABO genotyping, because it establishes within the limits of the number of samples tested that ABO genotypes can be assessed directly by non‐allele specific PCR amplification and restriction enzyme analysis.
Frozen Platelet Plates for Platelet Antibody Detection and Cross‐Match
Authors: Porretti L. , Marangoni F. , Rebulla P. , Sirchia G.
Published: March 2009

We performed 2 studies aimed at developing a frozen platelet panel suitable for platelet cross‐matching. The stability of the most important platelet membrane glycoproteins and the reactivity of antigens of the human platelet antigen (HPA) and of the human leukocyte antigen (HLA) systems were evaluated with the platelet suspension immunofluorescence test (PSIFT) in a panel of platelets frozen in microplates with 6% dimethylsulfoxide. In study No. 1 we evaluated platelet reaction with a broad‐spectrum weak anti‐HLA and a potent anti‐HPA‐1a antiserum and the expression of glycoproteins Ib and IIb/IIIa complex on platelet membrane before freezing and after 0.5, 1, 2, 3, 4, 5, 6 and 12 months of storage at ‐80°C. In study No. 2 we examined platelet reactivity with anti‐HPA‐1b, ‐HPA‐2a, ‐HPA‐3a, ‐HLA‐A2, ‐HLA‐A3 of platelets stored frozen for 12 months in parallel with fresh platelets from the same donors. Study No. 1 showed that glycoprotein expression was stable and that the weak anti‐HLA and the potent anti‐HPA‐1a antibodies were clearly detected during 12 months at ‐80°C. Of the 35 paired PSIFT performed in study No. 2 with fresh and frozen/thawed platelets incubated with anti‐HPA‐1b, ‐HPA‐2a, ‐HPA‐3a ‐HLA‐A2, ‐HLA‐A3 antisera and AB serum, concordant reactions were obtained in all cases with the exception of 1 case of HLA‐A3‐positive platelets incubated with anti‐HLA‐A3 antiserum, that was reactive with frozen/thawed platelets but nonreactive with fresh platelets from the same donor. The discrepant finding obtained with fresh platelets from 1 donor could be due to the well‐known variable and weak association of HLA antigens to platelet membrane. We conclude that frozen platelet plates can be stored and used for at least 12 months for detecting platelet‐reactive antibodies in patients' sera.
Clinical Significance of Anti‐Ata
Authors: Ramsey G. , Sherman L.A. , Zimmer A.M. , Spies W.G. , Sharma L. , Arndt P. , Garratty G.
Published: March 2009

Ata is a high‐frequency red blood cell (RBC) antigen. Anti‐Ata has been reported in rare At(a–) black subjects. We report two cases of anti‐Ata. A clinically significant anti‐Ata was found in a 26‐year‐old black woman with systemic lupus erythematosus. The patient had a transfusion reaction with chills and nausea during a RBC survival study, and 95% of the radiolabeled At(a+) RBCs were destroyed within 3 h. A concurrently performed monocyte monolayer assay was strongly reactive. Anti‐Ata thus can cause rapid hemolysis of transfused RBCs, but At(a–) donor units are extremely scarce in rare donor registries. A second patient at our hospital had anti‐Ata which did not affect her newborn. She also had autoimmune disease, insulin‐dependent diabetes mellitus.
Severe Hemolytic Disease of the Newborn due to Anti‐Jsb
Authors: Gordon Michael C. , Kennedy Melanie S. , O'Shaughnessy Richard W. , Waheed Abdul
Published: March 2009

Anti‐Jsb has not been reported to cause severe hemolytic disease of the newborn requiring intrauterine evaluation and treatment. We now report on such a patient with high‐titered anti‐Jsb causing fetal hydrops with demise in one pregnancy, followed by a pregnancy treated with multiple intrauterine fetal transfusions.
Description of magnetic resonance imaging‐derived enhancement variables in pathologically confirmed prostate cancer and normal peripheral zone regions
Authors: Zelhof Bashar , Lowry Martin , Rodrigues Greta , Kraus Sigurd , Turnbull Lindsay
Published: March 2009

OBJECTIVE

To assess the use of a semiquantitative analysis of dynamic contrast‐enhanced (DCE) magnetic resonance imaging (MRI) to produce indices for enhancement curves that might enable differentiation between malignant prostatic lesions and normal peripheral zone (PZ).

PATIENTS AND METHODS

Fifty‐two patients scheduled for radical prostatectomy underwent DCE‐MRI before surgery using a 3 T scanner. The DCE images were used to generate variables from changes in signal intensity for pathologically confirmed malignant areas and the normal PZ, using whole‐mounted pathology specimens as a reference to delineate regions of interest (ROI). These variables included maximum enhancement index (MaxEI), time to MaxEI at 30 s, the initial and final slopes of signal intensity change, and the area under curve. A threshold value for each DCE variable was identified, and the sensitivity and specificity were obtained.

RESULTS

Malignant lesions had a 56% higher MaxEI than normal PZ and took half the time to reach MaxEI (P < 0.001). Hence, at 30 s, cancer lesions have double the mean (sd) EI than normal PZ, of 2.22 (1.04) vs 1.04 (0.51), respectively. Tumours showed significant washout of contrast medium, which was reflected in the final slope of the curve being negative, as opposed to positive for normal PZ. The combined data of DCE variables, using a logistic regression test, gave a mean (95% confidence interval) sensitivity and specificity of 89 (81–96)% and 90 (83–97)%, respectively.

CONCLUSION

This technique provides good discrimination of malignant lesions that might enable accurate localisation of the lesion. It is a simple, semiquantitive, noninvasive method that reflects the unusual vascular characteristics of newly formed microvessels and the changes in the interstitium that occur in prostate cancer.
Safety Assessment Using Imprecise Reliability for Corrosion‐Damaged Structures
Authors: Choi Hyun‐Ho , Seo Jongwon
Published: March 2009

Abstract:  Safety assessment procedure including imprecise reliability for corrosion‐damaged structures is proposed in this article. The proposed procedure may be used for the safety assessment of corroded structures when the data for safety analysis are insufficient or when a decision for safety management is urgently required. In case exact measurement such as nondestructive test cannot be utilized to assess a corroded point for any reason, expert's estimation with simple visual inspection based on the data of nondestructive test at near point can be used. The system reliability‐based safety assessment may then be performed approximately but rationally by simply converting the qualitative expert's estimation to quantitative terms in this proposed procedure. Moreover, the approximate safety expressed as a safety range could also be evaluated effectively in hazardous environments such as typhoon where data collection for deterioration is very difficult. The proposed safety assessment procedure using the imprecise reliability was also applied to a cable‐stayed bridge in Korea to demonstrate its effectiveness and applicability.