bluuebunny/crossref_metadata_2025_split huggingface dataset:
Solid‐Phase Immunoassays for HCV Antibodies in Gamimune® N
Authors: Schreiner Clara , Musmanno Lisa , Kuenzi Mark , Torres Juan 
Published: December 2004

Abstract Background and objectives: Occasional false‐positive results for the presence of HCV antibodies occur when testing pH 4 Gamimune® N immune globulin intravenous (human) (IGIV) using commercially available immunoassay kits. We investigated the causes. Materials and methods: EIA test results were compared with Nile red hydrophobic dye binding. Results: There was a strict correlation of reactive EIA results with a larger relative proportion of molecules in a low pH‐dependent hydrophobic conformation. Conversely, the greater the hydrophilic contribution to the population of IgG molecules (favored at neutral pH), the more nonreactive the EIA test results became for the same lots. An IGIV test preparation procedure consisting of pH neutralization followed by 48‐hour incubation at 37°C decreased positive test result outcomes in four different immunoassay formats by shifting the IgG population to a more hydrophilic conformation. Conclusion: The hydrophobic conformation of IgG favored at low pH appears to give rise to significant levels of nonspecific binding in solid phase immunoassays.
Epidemiological and Clinical Aspects of Hepatitis G Virus Infection in Blood Donors and Immunocompromised Recipients of HGV‐Contaminated Blood
Authors: Heuft H. G. , Berg T. , Schreier E. , Künkel U. , Tacke M. , Schwella N. , Hopf U. , Salama A. , Huhn D. 
Published: December 2004

Abstract Background and objectives: The infectiousness and clinical relevance of the newly discovered blood‐borne Flaviviridae‐like agent, termed hepatitis G virus (HGV), are not well understood. Materials and methods: Twenty‐three transfusion recipients of two HGV‐affected long‐term blood donors were studied for HGV genome and antibodies to the putative envelope 2 glycoprotein (anti‐E2) of HGV. Nine recipients had nonhematological disorders and 14 suffered from severe hematological diseases and 7 of them received allogeneic bone marrow or blood stem cell transplantation. The molecular epidemiology of the observed HGV infection was studied by direct sequencing of parts of the 5′‐noncoding region, NS3, and NS5 region of HGV in the 2 long‐term donors and in their 6 recipients who became HGV RNA positive. Additionally, 549 individuals – homologous (n = 254) and autologous blood donors (n = 202), and medical staff (n = 89) – were investigated for the presence of HGV RNA. Results: HGV RNA in serum was found in 15 of the 23 (65%) transfusion reciients with known exposure of HGV‐contaminated blood. Seven of the remaining 8 recipients showed only an anti‐E2 response, indicating previous HGV infection with spontaneous clearance of the virus. In one recipient neither HGV RNA nor anti‐E2 could be detected. Molecular evidence for HGV transmission by the 2 donors was found in 3 of the 6 recipients studied. The alanine aminotransferase levels were not significantly different in the HGV RNA positive and negative recipients, and none of the 23 recipients developed posttransfusion hepatitis. Persistent HGV infection was observed especially in recipients with severe hematological disorders or in those in whome intensive immunosuppressive treatment was necessary. Of the 549 individuals studied, 10 (1.8%) were healthy carriers of HGV RNA. Conclusion: The persistence of transfusion‐acquired HGV infection is not associated with acute or chronic hepatitis, but may be influenced by the recipient's underlying disease.
Quality and Safety of Platelet Apheresis Concentrates Produced with a New Leukocyte Reduction System
Authors: Riggert J. , Humpe A. , Simson G. , Köhler M. 
Published: December 2004

Abstract Objectives: Contaminating white blood cells (WBC) in apheresis platelet concentrates (PC) can cause a variety of adverse effects after platelet transfusion. To obtain PCs with low WBC contamination, a new leukoreduction system (LRS) utilizing ‘fluidized particle bed’ technology has recently been introduced. Methods: We prospectively examined the effect of LRS apheresis on the donor, the quality of the resulting PCs (n = 120), and the platelet increment in the corresponding recipients. Conventionally prepared apheresis PCs served as control group (n = 27). Platelet glycoproteins were examined by flow cytometry. Results: In LRS apheresis, we observed no serious adverse effects on the donors, but the postdonation absolute lymphocyte counts were reduced from 1,787±505/μl to 1,405±383/μl (p<0.001). Comparable results were seen in non‐LRS donors. The collection efficiency of the LRS procedures was 50.0±7.6%, resulting in a yield of 4.3±1.0 × 1011 platelets/PC. In flow cytometry, platelet glycoproteins in LRS PCs were not elevated: mean fluorescence of CD62 (6±4) or CD63 (9±3) in comparison with non‐LRS PCs (mean fluorescence of CD62: 7±4, CD63: 8±3). Median leukocyte contamination of the LRS PCs was 0.41 × 105 (range 0.07–8.5) WBCs/unit. In 43 recipients, the 24‐hour corrected count increments after transfusion of LRS PCs (12,530±8,761) were essentially the same as those of 20 recipients of non‐LRS PCs (13,133±9,812; p = 0.75). Conclusions: LRS apheresis appears to be a safe procedure, which produced effective PCs with few contaminating leukocytes. With new apheresis technology, filtration of PCs may become superfluous.
Evaluation of a Platelet Apheresis Technique for the Preparation of Leukocyte‐Reduced Platelet Concentrates
Authors: Zingsem Jürgen , Glaser Anke , Weisbach Volker , Zimmermann Robert , Neidhardt Bernd , van Waeg Geert , Eckstein Reinhold 
Published: December 2004

Abstract Objectives: Reduction of the white blood cell (WBC) contamination in platelet concentrates (PC) protects patients from the immunological and infectious side effects of platelet transfusion caused by WBC. This can be done either by filtration of the PC or by improved apheresis techniques that yield WBC‐poor preparations. Methods: To evaluate an improved technique for platelet collection, we carried out 201 separations in 89 healthy cytapheresis donors using the new COBE Spectra leukoreduction system (LRS) and compared the results with those of standard dual‐needle separations obtained with the same cell separator. Results: A small but statistically significant difference was found in platelet collection efficiency in comparison with the standard non‐LRS software procedures (LRS: 52.6 vs. 56.3% for the reference). However, median WBC contamination was only 0.01 × 106 WBC per LRS product. This significant (p<0.0005) improvement corresponds to a 10‐fold reduction of WBC as compared with the standard dual‐needle technique. Conclusions: The COBE Spectra LRS system produced PCs with a platelet collection efficiency nearly equal to previous techniques and with a residual WBC content satisfying even the most stringent criteria for WBC‐depleted blood components. As this purity is achieved without important platelet loss, conventional fiber filtration no longer seems necessary in this kind of PC.
A Comparative Evaluation of the Sensitivity of Five Anti‐Hepatitis C Virus Immunoblot Assays
Authors: Couroucé A. M. , Noel L. , Barin F. , Elghouzzi M. H. , Lunel F. , North M. L. , Smilovici W. 
Published: December 2004

Abstract Background and Objectives: In order to compare sensitivity, five anti‐hepatitis C virus (HCV) immunoblot assays were tested (Deciscan plus, Inno‐Lia III, Matrix, Murex Western blot and RIBA‐3). Materials and Methods: The test panel (50 samples for each assay) included 6 anti‐HCV‐negative samples and 44 samples from 36 HCV‐infected subjects. Results: There were minor differences in core reactivity among the assays. The smallest number of NS3‐reactive results occurred with the Murex and Matrix assays, and the smallest number of NS4 reactives with RIBA‐3 and Matrix. Among the 20 discrepant results for NS5 there was one clear false‐negative with Inno‐Lia. Only 28 of the 50 samples of the panel gave the same results in all the assays: 5 negatives and 23 positives. One of the 6 negative samples was indeterminate in 3 assays. Eighteen of the 21 other divergent results were interpreted as either indeterminate or positive, a common reactivity being exhibited by all 5 assays. The most important discrepancies occurred on 3 HCV‐RNA‐positive samples which came up negative in some assays: 2 samples with isolated NS3 reactivity were negative by Matrix and Murex Western blot, 1 of them being also negative by Inno‐Lia III; another sample was negative by RIBA‐3 and Matrix due to weak signals (<1) on core and NS3 proteins, which did not exceed 1+ with the other assays. Conclusions: With more uniform criteria for interpretation, the results would have been less divergent. Some assays should improve their sensitivity to the NS3 protein.
Evaluation of the Performance of Brazilian Blood Banks in Testing for Chagas' Disease
Authors: Saéz‐Alquézar Amadeo , Otani Márcia M. , Sabino Ester C. , Ribeiro‐dos‐Santos Gabriela , Salles Nanci , Chamone Dalton F. 
Published: December 2004

Abstract Background and Objectives: Due to the low sensitivity and reproducibility of available tests, in 1989 it became mandatory for all Brazilian blood donors to be screened for Chagas' disease by at least two serological techniques. In 1994 the Brazilian Ministry of health launched a program to systematically evaluate the quality of serological screening for the detection of blood‐transmissible diseases as performed by public blood banks. Methods: A blind panel containing positive samples for blood‐transmissible disease was distributed to 57 major public blood banks in four sequential programs. Results: The ELISA test was chosen by the majority of the blood banks. There were 64 (3.7%) false‐negative results, 49 produced by banks using indirect hemagglutination. Since most blood banks screened with more than one test for Chagas, only 8 samples were actually missed, of which 3 were by banks using only one test. Conclusion: Our data show a clear improvement in performance of Brazilian blood banks testing for Chagas' disease.
Purification of α1 Proteinase Inhibitor from Human Plasma Fraction IV‐1 by Ion Exchange Chromatography
Authors: Chen Sharon X. , Hammond David J. , Lang John M. , Lebing Wytold R. 
Published: December 2004

Abstract Background and Objectives: α‐proteinase inhibitor (PI) protects the lungs from proteolytic damage caused by elastase and can be used to treat congenital emphysema. We describe an improved method of purification of α1 PI from redissolved fraction IV‐1 paste. Materials and Methods: The process used dimethylaminoethy1 anion exchange chromatography, sulfopropyl cation exchange chromatography, virus inactivation by dry heat, and tri‐n‐butyl‐phosphate/cholate treatment, followed by a second strong cation exchange chromatography. Optimizations of loading conditions for ion exchange chromatography at small scale (20–60 ml of suspension) are described. Virus inactivation was adjusted to provide the best yield of α1 PI consistent with effective inactivation. The process has been effectively scaled up. Results: The final product was approximately 90% pure by SDS‐PAGE, with a 60–70% yield from starting fraction IV‐I paste. The process has been characterized by methods including nonreduced SDS‐PAGE, α1 PI inhibition assay, and biuret protein assay. Conclusion: The method described is an effective way of preparing large quantities of α1 PI from fractionated plasma.
Different Alleles Cause an Imbalance in A2 and A2B Phenotypes of the ABO Blood Group
Authors: Ogasawara Kenichi , Yabe Ryuichi , Uchikawa Makoto , Bannai Makoto , Nakata Kenichi , Takenaka Michiko , Takahashi Yuji , Juji Takeo , Tokunaga Katsushi 
Published: December 2004

Abstract Background and Objectives: In several populations, including the Japanese, the frequency of the A2B phenotype is significantly higher than expected based on the A2 phenotype frequency. To understand the genetic basis of this ‘excess’ of A2B, we examined ABO alleles in individuals with A2‐related phenotypes. Materials and Methods: ABO alleles were identified by means of polymerase chain reaction single‐strand conformation polymorphism (SSCP) and nucleotide sequence analyses. Results: The frequencies of A2‐related alleles (*A105, *A106, *A107, *A111 and *R101) were clearly different between the A2 and A2B phenotypes. In particular, a putative recombinant allele, *R101, was uncommon in the A2 but common in the A2B phenotype individuals. This allele was also detected in 4 of 401 (1%) unrelated A1 phenotype (AO genotype) individuals. Conclusion: *R101 is presumably expressed as phenotype A1 in *R101/*O heterozygous individuals, but as phenotype A2 in *R101/*B heterozygotes, thus giving rise to a high A2B phenotype frequency.
Serum Affinity Chromatography for the Detection of lgG Alloantibodies in a Patient with High‐Titer lgM Cold Agglutinins
Authors: Stahl D. , Kreft H. , Hack H. , Schraven B. , Roelcke D. 
Published: December 2004

Abstract Background and Objectives: High‐titer cold autoagglutinins (CA) interfere with the detection of alloantibodies against red‐blood‐cell antigens. The diagnostic procedure is extremely difficult, especially under pressure of time. It was the purpose of the study to find out whether quantitative IgG purification from serum of plasma by affinity chromatography facilitates the detection of IgG alloantibodies in the presence of high‐titer IgM CA. Materials and Methods: Serum: IgM‐ϰ CA, 0°C titer = 65,000; five anti‐IgG reactive alloantibodies. Affinity chromatography: HiTrap protein G 1‐ml affinity column (Pharmacia Biotech, Uppsala, Sweden). Results: All IgG alloantibodies were detectable in the eluate after affinity chromatography without interference of IgM CA. Conclusion: IgG alloantibodies in the presence of high‐titer IgM CA can be easily and rapidly detected under routine conditions by quantitative IgG purification from serum or plasma by affinity chromatography.
Transfusion Practices in Primary Total Joint Replacements in Finland
Authors: Capraro Leena 
Published: December 2004

Abstract Background and Objectives: A multicenter survey was carried out in 11 Finnish hospitals to determine the prevailing transfusion practices in orthopedic surgery. Materials and Methods: The records of 1,161 patients who had undergone primary total hip (THR) or knee replacement (TKR) were reviewed. Results: Allogeneic red cells (RBC) were administered to 92% of the THR patients and to 84% of the patients undergoing TKR. Significant interhospital variation in RBC transfusion, from 2.4 to 4.1 units per patient in THR and from 1.8 to 2.8 units in TKR, was detected after adjustment for clinical and laboratory findings. Conclusion: The use of RBCs in orthopedic surgery was generous compared with reports from other European countries and the US. Each hospital's individual transfusion policy seemed to be a strong determinant of allogeneic blood use.
Point Mutations and Deletion Responsible for the Bombay H null and the Reunion H weak Blood Groups
Authors: Fernandez‐Mateos Pilar , Cailleau Anne , Henry Stephen , Costache Marieta , Elmgren Anders , Svensson Lola , Larson Göran , Samuelsson Bo E. , Oriol Rafael , Mollicone Rosella 
Published: December 2004

Abstract Objective: Definition of the molecular basis of the Reunion and the Bombay red cell and salivary H‐deficient phenotypes. Methods: Sequence and expression of FUT1 and FUT2 genes from H‐deficient individuals. Family segregation analysis of the mutations responsible for the fucosyltransferase defects of H, secretor and Lewis systems. Results: The Indian red cell H null Bombay phenotype depends on a new mutation of the FUT1 gene. T725→G changing Leu242→Arg. Their salivary nonsecretor phenotype is secondary to a complete deletion of the FUT2 gene. The red cell H weak Reunion phenotype depends on another new mutation of FUT1, C349→T which induces a change of His117→Tyr. Their salivary nonsecretor phenotype is due to the known Caucasian inactivating mutation G428→A. Conclusion: Single prevalent FUT1 and FUT2 point mutations and a deletion are responsible for the Indian Bombay H null and the Reunion H weak phenotypes found on Reunion island. This is in contrast with other H‐deficient phenotypes where sporadic nonprevalent inactivating mutations are the rule.
Different Genotypes Causing Indiscernible Patterns of A Expression on A el Red Blood Cells as Visualized by Scanning Immunogold Electron Microscopy
Authors: Hansen Torjørn , Namork Ellen , Olsson Martin L. , Alan Chester M , Heier Hans Erik 
Published: December 2004

Abstract Background and Objectives: The published sequence of the weak A subgroup Ael gene from Swedish individuals showed a G insertion in exon VII, causing a frameshift at ccdon 268 (the A01 gene has 353 codons). We wished to sequence exons VI and VII of two Norwegian Ael individuals and compare the expression of A substance on RBC from different A el individuals. Materials and Methods: Exon VI and VII were amplified by PCR, cloned in M13 and sequenced. A structure expression on Ael RBC was studied by the immunogold technique. Results: In contrast to the Swedish Ael individuals, the two Norwegians had consensus A01 sequences in exon VI and VII. However, the patterns of A expression were indiscernible from the Swedish cases as visualized by immunogold labeling in SEM. In both cases, a few (1–2%) RBC were very strongly labeled, some were weakly labeled and the majority (95%) were unlabeled. Conclusion: Although some Ael individuals have an inserted nucleotide in exon VII of the ABO gene, others have consensus A01 sequence in exon VI and VII. However, we could not find any differences in phenotype by immunogold labeling in SEM.
RHD Gene Polymorphisms among RhD‐Negative Chinese in Taiwan
Authors: Sun Chien‐Feng , Chou Cheng‐Sheng , Lai Nan‐Chang , Wang Wei‐Ting 
Published: December 2004

Abstract Background and Objective: The rare occurrence of anti‐D‐associated hemolytic disease of the newborn among Chinese is attributable in part to the existence of the weak D phenotype Del among apparently RhD‐negative individuals. While exciting advances in the molecular genetics of the Rh blood group have been noted in recent years, the genomic structure of the Del phenotype has seldom been studied in the literature. We try to explore the genomic structure of the RhD gene among apparently Rh‐negative Chinese in Taiwan in this study. Methods: Genomic DNA from 230 samples of apparently RhD‐negative Chinese was studied using four polymerase chain reaction (PCR)‐based RhD typing methods. These PCR methods amplified RHD and RHCE genes at exons 4, 5, 7 and 10. All nuclieotides responsible for exofacial amino acid differences between RhD and RhCeEe peptides, including amino acids 169, 170, 172, 223, 226, 233, 238, 350, 353, and 354, were contained in these amplified DNA segments. Southern blot analysis using RHD cDNA fragments as probes was performed. Results: According to the serological study, 155 samples (67.4%) were genuinely RhD‐negative and 75 samples (32.6%) were of the Del phenotype. Successful amplifications for RHD sequences were possible in all 75 Del samples using four PCR methods. Apparently, all Del individuals carried an intact RHD gene. While 145 individuals of 155 genuinely Rh‐negative (63.0% of apparently RhD‐negative individuals) had total deletion of their RHD genes, 10 individuals (4.3% of apparently RhD‐negative individuals) were shown to have a preserved 3′ noncoding region of the RHD exon 10 and a gross deletion of RHD exons 4–10. Conclusions: Three classes of RhD‐negative polymorphisms among Chinese in Taiwan were observed. These included Del with grossly intact RHD and weak RhD expression, genuinely RhD‐negative with total deletion of the RHD gene. A molecular study is warranted to clarify the mechanism responsible for the weak RHD gene expression in Del individuals.
Characterization of a Macaque Anti‐Rh17‐Like Monoclonal Antibody
Authors: Blancher A. , Roubinet F. , Reid M.E. , Socha W.W. , Bailly P. , Bénard P. 
Published: December 2004

AbstractIn order to produce macaque monoclonal antibodies (mAbs) against human red blood cell (RBC) antigens, macaques were immunized with human and gorilla RBCs and their spleen lymphocytes were fused with man‐mouse heteromyeloma cells. One macaque‐mouse heterohybridoma produced a macaque IgGχ (Cyn2‐4D5) which agglutinated all human RBCs but not rare human variants Dc–, D––, and Rhnull. Thus, Cyn2‐4D5 exhibited RH17‐like reactivity. The specificity of Cyn2‐4D5 for RHCE‐encoded polypeptides was confirmed by specific immunoprecipitation of RhcE and RhCe polypeptides from human RBCs and the absence of immunoprecipitation of the RhD polypeptides extracted from D–RBCs. This study demonstrates that it is possible to produce macaque mAbs against human RBC blood group antigens.
The ELO Blood Group Polymorphism Is Located in the Putative First Extracellular Loop of Human Erythrocyte Band 3
Authors: Zelinski Teresa , Punter Fiona , McManus Kirk , Coghlan Gail 
Published: December 2004

Abstract Background and Objectives: The low incidence red cell antigen ELO (700.51) has been excluded from 13 of 23 established blood group systems. Information relative to the Diego system has not been reported. To investigate a possible association between ELO and the gene controlling Diego blood group polymorphisms, we undertook molecular studies of AE1 (anion exchanger 1 gene). Materials and Methods: DNA from a family segregating for ELO and from the original ELO proposita was amplified by PCR using intronic primers flanking exons 11‐20 of AE1. Subsequently, single‐strand conformational polymorphism (SSCP) analysis and DNA sequencing were conducted. Results: We have observed an exon 12 SSCP in all ELO+ individuals tested. This SSCP is the result of a C→T mutation in exon 12 of AE1 which leads to an Arg432→ Trp amino acid substitution in the putative first extracellular loop of band 3 and thereby accounts for the Diego blood group system polymorphism known as ELO. Conclusions: In light of these findings, the International Society of Blood Transfusion Working Party on Terminology for Red Cell Surface Antigens has designated ELO as a member of the Diego blood group system (010008 or DI18) and rendered the previous numerical designation (700.51) obsolete.
A Simple Strategy to Look Back on Posttransfusion Hepatitis B in a Multitransfused Patient
Authors: Jongerius J. M. , van der Poel C. L. , van Leeuwen C. F. 
Published: December 2004

Abstract Background and Objectives: In January 1996, a case of hepatitis B virus (HBV) seroconversion in a multitransfused patient was reported to the blood bank. From March through October 1995, the patient had received 23 units of red cells and 30 units of pooled platelet concentrates, encompassing an exposure to a total of 200 whole blood donations. Materials and Methods: In order to trace hepatitis B surface antigen (HBsAg)‐negative but HBV‐infectious blood donation(s), we tested samples of the donors obtained ≥3 months after the implicated donations for anti‐HBc (Corezyme EIA, Abbott). From 172/200 donors, archived samples of subsequent donations were available for this purpose. The remaining 28 donors were reinvited to the blood bank to obtain an additional blood sample for anti‐HBc testing. Results: 1/200 follow‐up donor samples was anti‐HBc‐positive. Retrospective testing of the implicated HBsAg‐negative blood donation of this donor revealed anti‐HBc‐negative and HBV‐DNA‐positive results. The patient was transfused with the platelets of the HBV‐infectious donation. On looking back, the other blood products prepared from this HBV‐infectious donation caused posttransfusion HBV infection (PT‐HBV) in 2 additional patients. Conclusion: Anti‐HBc testing on mainly archived follow‐up samples of 200 donors implicated in PT‐HBV was a rapid, simple, cost‐effective and donor friendly method to identify an infectious but HBsAg‐negative, anti‐HBc‐negative and HBV‐DNA PCR‐positive blood donation. Routine anti‐HBc screening would not have prevented this HBV transmission.