Context.— Serologic tests on automated immunology analyzers are increasingly used to monitor acquired immunity against SARS-CoV-2. The heterogeneity of assays raises concerns about their diagnostic performance and comparability. Objective.— To test sera from formerly infected individuals for SARS-CoV-2 antibodies by using 6 automated serology assays and a pseudoneutralization test (PNT). Design.— Six SARS-CoV-2 serology assays were used to assess 954 samples collected during a 12-month period from 315 COVID-19 convalescents. The tests determined either antibodies against the viral nucleocapsid (anti-NC) or spike protein (anti-S). Two assays did not distinguish between antibody classes, whereas the others selectively measured immunoglobulin G (IgG) antibodies. PNT was used to detect the presence of neutralizing antibodies. Results.— Comparison of qualitative results showed only slight to moderate concordance between the assays (Cohen κ < 0.57). Significant correlations (P < .001) were observed between the antibody titers from all quantitative assays. However, titer changes were not detected equally. A total anti-S assay measured an increase in 128 of 172 cases (74%) of a suitable subset, whereas all IgG anti-S tests reported decreases in at least 118 (69%). Regarding the PNT results, diagnostic sensitivities of 89% or greater were achieved with positive predictive values of at least 93%. In contrast, specificity changed substantially over time, varying from 20% to 100%. Conclusions.— Comparability of serologic SARS-CoV-2 antibody tests is rather poor. Owing to different diagnostic specificities, the tested assays were not equally capable of capturing changes in antibody titers. However, with thoroughly validated cutoffs, IgG-selective anti-S assays are a reliable surrogate test for SARS-CoV-2 neutralizing antibodies in former COVID-19 patients.

BACKGROUND:Blood group A and B antigens are synthesized by glycosyltransferases regulated by a complex molecular genetic background. A multibase deletion in the ABO gene was identified in two related blood donors. To define its hereditary character and to evaluate genotype-phenotype associations, a detailed study including 30 family members was conducted. METHODS AND MATERIALS:ABO phenotyping was performed with agglutination techniques and adsorption-elution tests. The secretor status was determined. Allele-specific sequencing of ABO and genotyping of family members by a mutation-specific polymerase chain reaction were carried out. Functional analysis included cloning of complementary DNA and transfection experiments in HeLa cells. The antigen expression was investigated by flow cytometry and adsorption-elution method. RESULTS:Sequencing analysis revealed a 24-bp deletion in Exon 5 and the adjacent intronic region of ABO. The alteration was inherited by 16 family members. Nine of them being heterozygous for the mutated allele failed to express A antigen on their erythrocytes as found by routine typing. In particular samples, however, adsorption-elution studies indicated inconclusive results. HeLa cells transfected with aberrant gene transcripts did not express blood group antigen A. CONCLUSION:The variation causes defects in messenger RNA splicing, most likely inactivating the transferase as observed by serological typing and in vitro expression analysis. These data suggest a novel mechanism associated with blood group O and extend the knowledge of exceptionally rare ABO splice site mutations and deletions. With increased understanding of the molecular bases of ABO, the diagnostics may be further enhanced to ensure the safest possible use of the blood supply.

Since the emergence of SARS-CoV-2 numerous antibody assays have become available, demonstrating different performance characteristics. This study focused on a quantitative correlation between different commercial assays and a neutralization test (NT). Comparative data is needed as a basis for the production of convalescent plasma and potential interpretations COVID-19 immunity. Sera of 100 SARS-CoV-2 convalescent plasma donors were collected and SARS-CoV-2 antibodies were characterized using three different IgG-ELISAs (EUROIMMUN IgG and NCP-IgG ELISA, Wantai ELISA), two CLIA (Elecsys, LIAISON) and two lateral flow tests (MEDsan IgM/IgG-Rapid-Test, Wantai Rapid Test) and subsequently correlated to neutralization titers. The Wantai ELISA and the Elecsys provide the highest sensitivities in this sample (98 and 95 percent respectively). Titers with the best overall quantitative correlation to the NT titer were obtained with the Euroimmun IgG ELISA assay (Rho=0.759) and the Wantai ELISA assay (Rho=0.729). An infection without fever and negative or weakly positive reactions in the Wantai Rapid test were negative predictive factors for NT titers >1:200 (negative predictive value of 92 % and 92 % respectively, combination of both 100 %). The Wantai ELISA titer could be a suitable substitute for NT. An adequate pooling strategy of plasma units additionally could compensate deviations of individual antibody titers.

Background: Besides anemia, iron deficiency may cause more subtle symptoms, including the restless legs syndrome (RLS), the chronic fatigue syndrome (CFS) or sleeping disorders. Objective: The aim of this pre-planned secondary analysis of the IronWoMan randomized controlled trial (RCT) was to compare the frequency and severity of symptoms associated with iron deficiency before and after (intravenous or oral) iron supplementation in iron deficient blood donors. Methods/Design: Prospective, randomized, controlled, single-centre trial. (ClinicalTrials.gov: NCT01787526). Setting: Tertiary care center in Graz, Austria. Participants: 176 (138 female and 38 male) whole-blood and platelet apheresis donors aged ≥ 18 and ≤ 65 years with iron deficiency (ferritin ≤ 30ng/mL at the time of blood donation). Interventions: Intravenous iron (1 g ferric carboxymaltose, n = 86) or oral iron supplementation (10 g iron fumarate, 100 capsules, n = 90). Measurements: Clinical symptoms were evaluated by a survey before iron therapy (visit 0, V0) and after 8–12 weeks (visit 1, V1), including questions about symptoms of restless legs syndrome (RLS), chronic fatigue syndrome (CFS), sleeping disorders, quality of life and symptoms like headaches, dyspnoea, dizziness, palpitations, pica and trophic changes in fingernails or hair. Results: We found a significant improvement in the severity of symptoms for RLS, fatigue and sleep quality (p < 0.001). Furthermore, a significant decrease in headaches, dyspnoea, dizziness and palpitations was reported (p < 0.05). There was no difference between the type of iron supplementation (intravenous versus oral) and clinical outcome data. Conclusion: Iron supplementation in iron-deficient blood donors may be an effective strategy to improve symptoms related to iron deficiency and the wellbeing of blood donors.

Background: Routine ABO blood group typing for pre-transfusion testing of a male Austrian patient of Far Eastern origin showed discrepant results with an apparently weak blood group B phenotype and irregular anti-B. Materials and Methods: ABH phenotyping and cross-matching was done by standard serologic techniques and levels of H expression were determined by flow cytometry. ABO gene sequencing including regulatory regions as well as analysis of FUT1 (H), FUT2 (Secretor), and FUT3 (Lewis) were carried out. Results: While monoclonal ABO antigen typing indicated blood group O, weak agglutination reactions using polyclonal human anti-B and anti-AB were seen. In reverse typing at room temperature, the plasma was reactive with A1 and A2 RBCs and negative with B and O cells, whereas at 4°C, anti-B reactivity was found. The indirect anti-globulin cross-match of the patient’s plasma was positive with group B RBCs and negative with group O RBCs. Sequencing analysis showed the presence of ABO*B.01 (B114) allele and homozygosity for the FUT1 mutation c.551_552delAG. Flow cytometry demonstrated trace amounts of H antigen on the patient’s RBCs. Conclusion: While a functional B allele was found, analysis of FUT1 and FUT2 genes revealed the presence of a rare para-Bombay genotype OhB. Interestingly, no anti-H but irregular anti-B was found in the patient’s plasma, responsible for the positive cross-match with group B RBCs. Even though very rare and not reported for the European population, the presence of an H-deficient phenotype should be considered when investigating individuals with an unusual ABO blood group type.

INTRODUCTION:Frequent blood donation often leads to iron deficiency and even anemia but appropriate strategies for detection and prevention are currently not mandatory. At the Medical University of Graz, we conducted a single-center prospective clinical trial to compare oral and IV iron supplementation in iron deficient blood donors including Austrian regular whole blood and platelet apheresis donors. We aimed to determine the difference of transferrin saturation between the treatment groups 8-12 weeks iron administration besides other parameters of iron status and blood count. METHODS:176 healthy male and female blood donors with iron deficiency (ferritin ≤30 ng/mL) were randomized to either a single dose of IV ferric carboxymaltose (1000 mg, n = 86) or oral iron (II)fumarate (100 tablets of 100 mg [10 per week], n = 90). RESULTS:Between 2014 and 2016, 172 donors (137 women) completed the study; 4 in the oral group were lost to follow-up. At follow-up, median (IQR) transferrin saturation and ferritin were significantly higher in the intravenous group (27 [23-35]%, vs 21.0 [16-32]%; p < 0.001 and 105 [75-145] ng/mL vs 25 [17-34] ng/mL; p < 0.001, respectively) while median (IQR) hemoglobin levels were comparable (IV, 13.6 [13.0-14.4] g/dL vs oral, 13.6 [13.0-14.2] g/dL). The frequency of adverse effects was comparable (38% in both groups) and no serious adverse events occurred. CONCLUSIONS:A single dose of 1000 mg of intravenous iron is highly effective to counteract iatrogenic iron deficiency in blood donors. Oral iron appears to be an acceptable alternative. The assessment of body iron stores should play a key role in maintaining blood donors' health. This trial was registered at www.clinicaltrials.gov as NCT01787526 on February 8, 2013 and at www.clinicaltrialsregister.eu (EudraCT identifier: 2013-000327-14) on September 24, 2013.

s von Vorträgen und Postern* *Reihung alphabetisch nach Erstautor Regeneration von oralem Gewebe: Die Regulation von Vascular Endothelial Growth Factor, Sklerostin und Dickkopf-1 in parodontalen Fibroblasten H. Agis1, L. Hueber 1, M. Gruber 1, M. Silver 2, G. Watzek1, R. Gruber1 1Abteilung für Orale Chirurgie, Medizinische Universität Wien, und Austrian Cluster for Tissue Regeneration, Österreich; 2Harvard School of Dental Medicine, Boston, USA Die Gefäßneubildung ist essenziell für die Regeneration von Knochen und Weichgewebe. Es ist bekannt, dass die Reduktion des Sauerstoffpartialdrucks, also Hypoxie, die Entstehung neuer Gefäße auslöst. Ein rezenter Ansatz zur Förderung der Regeneration von Knochen und Weichgewebe ist daher die pharmakologische Simulation von Hypoxie durch so genannte „small molecules“. Die Wirkung dieser „small molecules“ auf parodontales Gewebe ist aber noch ungeklärt. Wir untersuchten daher die Wirkung von „small molecules“ in Bezug auf die Produktion von pro-angiogenem „vascular endothelial growth factor“ (VEGF) durch parodontale Fibroblasten. Es zeigt sich, dass „small molecules“ in vitro die VEGF-Produktion steigern können, ohne die inflammatorische Antwort substanziell zu modulieren. Unsere Untersuchungen über die Wirkung von Knochenersatzmaterial, welches mit „small molecules“ funktionalisiert wurde, zeigen, dass diese Prozessierung die Produktion von VEGF steigern kann. Ob diese pro-angiogene Wirkung der „small molecules“ zu einer Steigerung der Gewebsregeneration führt, ist Gegenstand präklinischer Untersuchungen. Sklerostin und Dickkopf-1 sind zentrale Regulatoren der Neubildung von Knochen, indem die beiden Faktoren den Wnt-Signalweg blockieren. Sklerostin und Dickkopf-1 sind möglicherweise auch an jenen Prozessen beteiligt, die den Erhalt der Zähne ermöglichen, aber auch pathophysiologische Wirkungen sind nicht auszuschließen. Die Frage nach der Regulation der Expression kann als erster Schritt zum Verständnis der Wirkung von Sklerostin und Dickkopf-1 angesehen werden. Basierend auf dieser Frage wurden parodontale Fibroblasten mit Wachstumsfaktoren und inflammatorischen Faktoren, die im Parodont vorkommen, inkubiert. Untersucht wurde die Produktion von Sklerostin und Dickkopf-1 auf mRNAund Proteinebene. Wir konnten zeigen, dass „transforming growth factor beta“ die Produktion von Sklerostin in parodontalen Fibroblasten sowohl auf mRNAals auch auf Proteinebene steigert. Weiters fanden wir, dass „tumor necrosis factor alpha“ die Produktion von Dickkopf-1 in parodontalen Fibroblasten sowohl auf mRNAals auch auf Proteinebene steigert. Zusammengefasst zeigen diese Daten, dass sowohl Sklerostin als auch Dickkopf-1 in parodontalen Zellen durch unterschiedliche Faktoren reguliert werden. Sklerostin und Dickkopf-1 könnten demnach unterschiedliche funktionelle Bedeutungen im Zahnhalteapparat haben. Diese Annahme ist insofern interessant, da zukünftige Therapien mit Anti-Sklerostinund Anti-Dickkopf-1-Antikörpern eine Auswirkung auf das Parodont haben könnten. Sclerostin and Its Association with Age, Gender, Body Composition, Physical Activity, and Bone Mineral Content in Healthy Adults K. Amrein1, S. Amrein 2, C. Schnedl1, C. Drexler3, H. P. Dimai1, H. Dobnig1, A. Tomaschitz1, T. R. Pieber1, A. Fahrleitner-Pammer1 1Division of Endocrinology and Metabolism, Department of Internal Medicine, 2Department of Anaesthesiology and Intensive Care Medicine, and 3University Clinic of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Austria Background Sclerostin is produced by osteocytes and inhibits bone formation through the Wnt/β-catenin signalling pathway. Only limited data is available on circulating sclerostin levels in healthy subjects. Therefore we aimed to evaluate the correlation between sclerostin and physical activity (PA), anthropometric and biochemical variables in healthy adult men and premenopausal women. Methods Serum sclerostin levels were measured in 161 healthy adults aged 19–64 years (mean 44 ± 10). Body composition and bone mineral density (BMD) were assessed with dual x-ray absorptiometry. Physical activity was recorded with a standardized questionnaire. Results A positive correlation between age and sclerostin in both men (r = 0.37; p < 0.001) and premenopausal women (r = 0.66; p < 0.001) was found. Men had significantly higher sclerostin levels than women (49.8 ± 17.6 vs 37.2 ± 15.2 pmol/l; p < 0.001). However, after adjustment for age, bone mineral content (BMC), PA, BMI, and renal function sclerostin levels did not differ (p = 0.543). Partial correlation analysis adjusted for age, gender, and kidney function revealed a significantly positive correlation between sclerostin levels and BMC, BMD, BMI, and android/gynoid fat and a significantly negative correlation with serum osteocalcin and calcium. The most physically active quartile had significantly lower sclerostin levels compared to the least active quartile in an univariate analysis. Conclusion In healthy adults, sclerostin serum levels correlate positively with age, Body Mass Index, and bone mineral content and negatively with osteocalcin and calcium. Further studies in larger populations are needed to confirm our findings and to better understand their clinical implications. Einfluss unterschiedlicher Thrombozytenpräparationen auf die osteogene Differenzierung M. Eichholzer1, M. Silver 2, G. Watzek1, H. Agis1, R. Gruber1 1Abteilung für Orale Chirurgie, Medizinische Universität Wien, und Austrian Cluster for Tissue Regeneration, Österreich; 2Harvard School of Dental Medicine, Boston, USA Die klinische Anwendung von „platelet-rich plasma“ (PRP) hat das allgemeine Interesse geweckt, die Funktion der Thrombozyten im Rahmen der Knochenregeneration besser zu verstehen. Die Knochenregeneration ist ein hochkonservierter, mehrstufiger Prozess, der auf dem Zusammenwirken von spezialisierten Zellen beruht. Hier kommt den knochenaufbauenden Osteoblasten eine zentrale Bedeutung zu. Um die Wirkung von Thrombozyten auf die Bildung der Osteoblasten besser zu verstehen, wurden In-vitro-Studien initiiert, in welFor personal use only. Not to be reproduced without permission of Krause & Pachernegg GmbH. J MINER STOFFWECHS 2011; 18 (SUPPLEMENTUM 3) 7 ÖGKM-Herbsttagung 2011 – Abstracts chen der Effekt von Thrombozytenpräparationen auf den Prozess der Osteoblastogenese untersucht wurde. Die Ergebnisse der vorliegenden Studien sind scheinbar widersprüchlich. Untersuchungen über den Einfluss von aktiviertem PRP zeigen, dass PRP die Entstehung von Osteoblasten fördert. Untersuchungen über den Einfluss von Wachstumsfaktoren aktivierter Thrombozyten („platelet-released supernatant“, PRS) können das hingegen nicht bestätigen. Eine Studie, die einen direkten Vergleich ermöglicht, liegt nicht vor. Ziel dieser Studie war es, die Wirkung von PRP und PRS auf die Osteoblastogenese direkt zu vergleichen. Hierzu wurden osteogene Zellen der Zelllinie MG63 und primäre osteogene Zellen mit folgenden 3 Präparationen stimuliert: (i) Überstände von PRP, welche sowohl Wachstumsfaktoren von Thrombozyten als auch Serumbestandteile enthalten; (ii) PRS, welches Wachstumsfaktoren von Thrombozyten enthält; (iii) Überstände von „fresh frozen plasma“ (FFP), welches Serumbestandteile enthält. Die Wirkung auf die osteogene Differenzierung der Zellen wurde mittels histochemischer Färbung der alkalischen Phosphatase bestimmt. Weiters wurde die Wirkung auf die Proliferation der Zellen mittels [3H]Thymidin-Einbau gemessen. Unsere Ergebnisse zeigen, dass Überstände von PRP sowie FFP, also die Präparationen, die Serumbestandteile enthalten, die osteogene Differenzierung von MG63 steigern können. PRS hingegen, welches keine Serumbestandteile enthält, führt nicht zu einer derartigen Steigerung. Ein ähnliches Bild zeigt sich auch bei primären osteogenen Zellen. Zusammengefasst weisen unsere Ergebnisse darauf hin, dass die Gegenwart von Serum in den Thrombozytenpräparationen den Effekt auf die osteogene Differenzierung beeinflussen kann. Diese Tatsache sollte bei der Wahl von In-vitro-Modellen zur Untersuchung der Wirkung von PRP und Thrombozyten auf osteogene Zellen berücksichtigt werden. Epigenetic Regulation of the Calcium-Sensing

The authors have disclosed no conflicts of interest.

TransfusionVolume 57, Issue 10 p. 2548-2549 LETTER TO THE EDITOR Iron deficiency in blood donors: perceptions and management among general practitioners and internists Karin Amrein MD, MSc, Karin Amrein MD, MSc karin.amrein@medunigraz.at Department of Internal Medicine, Division of Endocrinology and Diabetology, Medical University of Graz, Graz, AustriaSearch for more papers by this authorSusanne Macher MD, Susanne Macher MD Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorMartin Schröck MD, Martin Schröck MD Department of Internal Medicine, Division of Endocrinology and Diabetology, Medical University of Graz, Graz, Austria Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorPeter Schlenke Prof., MD, Peter Schlenke Prof., MD Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorCamilla Drexler MD, Camilla Drexler MD Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this author Karin Amrein MD, MSc, Karin Amrein MD, MSc karin.amrein@medunigraz.at Department of Internal Medicine, Division of Endocrinology and Diabetology, Medical University of Graz, Graz, AustriaSearch for more papers by this authorSusanne Macher MD, Susanne Macher MD Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorMartin Schröck MD, Martin Schröck MD Department of Internal Medicine, Division of Endocrinology and Diabetology, Medical University of Graz, Graz, Austria Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorPeter Schlenke Prof., MD, Peter Schlenke Prof., MD Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorCamilla Drexler MD, Camilla Drexler MD Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this author First published: 25 September 2017 https://doi.org/10.1111/trf.14297Citations: 2Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article.Citing Literature Volume57, Issue10October 2017Pages 2548-2549 RelatedInformation

About 2–3 % of the population participates in blood donation programmes. Each whole blood donation or ten apheresis donations cause a loss of 200–250 mg of iron. As a result, one of the most common risks of regular blood donors is iron deficiency. Although this has been known for decades, in most countries, iron status is currently not assessed or treated in this population. Premenopausal women are particularly affected, as they have lower iron reserves and higher daily requirements. Besides anaemia, iron deficiency may lead to fatigue and impaired cognitive and physical performance. Current iron preparations for intravenous administration are well tolerated and allow for application of large doses up to 1 g in one visit. Our hypothesis is that in blood donors with iron deficiency, intravenously administered iron is more efficient and as safe as oral iron supplementation. Since anaemia is one of the most frequent reasons for permanent or intermittent donor deferral, maintaining an iron-replete donor pool may help to prevent shortages in blood supply and to avoid iron deficiency-related comorbidities.

BACKGROUND: The presence of ABO subgroup alleles and unusual O alleles often is associated with discrepant serologic findings in ABO blood group typing. In the ABO gene of a Caucasian female and her daughters who had aberrant ABO phenotypes, a novel ABO O allele characterized by a large deletion that included two exons was identified.METHODS: ABO phenotypes were determined by standard agglutination tests and adsorption-elution studies. Allele-specific sequencing analyses of the ABO gene as well as messenger RNA transcripts were carried out.RESULTS: All three samples showed the same discrepant ABO blood typing results lacking A and B antigens, indicating Blood Group O, whereas anti-A1 and anti-A2 isoagglutinins were not detectable in reverse typing. Analyses of the ABO gene revealed a novel allele characterized by a deletion of 2169 base pairs, including sequences of Intron 1, Exon 2, Intron 2, Exon 3, and Intron 3. Exon 1 was directly joined to Exon 4 in the ABO transcript.CONCLUSION: Because the novel allele was associated with a well-described O allele, the absence of A-antigens in the inherited ABO subtype phenotype may be due to the identified mutation affecting the transmembrane-spanning domain of the encoded protein and impairing the transferase activity.

TransfusionVolume 55, Issue 6pt2 p. 1589-1590 NEW ALLELES AND ANTIGENS - SHORT REPORTS Novel ABO gene variants caused by missense mutations in Exon 7 leading to discrepant ABO blood typing results Eva M. Matzhold, Corresponding Author Eva M. Matzhold Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaAddress reprint requests to: Eva Maria Matzhold, Mag. Dr. scient. Med., Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Auenbruggerplatz 3, 8036 Graz, Austria; e-mail: [email protected].Search for more papers by this authorAndrea Wagner, Andrea Wagner Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorCamilla Drexler, Camilla Drexler Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorThomas Wagner, Thomas Wagner Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this author Eva M. Matzhold, Corresponding Author Eva M. Matzhold Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaAddress reprint requests to: Eva Maria Matzhold, Mag. Dr. scient. Med., Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Auenbruggerplatz 3, 8036 Graz, Austria; e-mail: [email protected].Search for more papers by this authorAndrea Wagner, Andrea Wagner Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorCamilla Drexler, Camilla Drexler Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this authorThomas Wagner, Thomas Wagner Department of Blood Group Serology and Transfusion Medicine, Medical University of Graz, Graz, AustriaSearch for more papers by this author First published: 09 April 2015 https://doi.org/10.1111/trf.13025Citations: 4Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL No abstract is available for this article.Citing Literature Volume55, Issue6pt2Special Issue: Immunohematology and Blood Group GenomicsJune 2015Pages 1589-1590 RelatedInformation

BackgroundPlatelet-reactive antibodies lead to thrombocytopenia and bleeding disorders, and diverse assays are used for their detection. In this retrospective analysis, the applicability of three different test systems was compared and antibody specificities were assessed. MethodsSera of 1,234 patients were tested with an enzyme-linked immunosorbent assay (ELISA; Lifecodes PAKPLUS (R) or PAK 12 (R), Gen-Probe) and a solid-phase assay (Capture-P Ready Screen (R), Immucor Inc.). In cases of suspected anti-HLA class I antibodies, a specific lymphocytotoxicity test (LCT, Bio-Rad (R)) was performed. ResultsPlatelet antibodies were detected in 366 of 1,234 samples (29.7%). In 70.3% concordant negative but only in 8.4% concordant positive results were obtained with both the methods; 185 of 1,053 in the solid-phase assay negative samples were positive in the ELISA (15.0%). In samples positive in both methods, most antibodies reacted against HLA class I antigens. Glycoprotein (GP) specific platelet antibodies, mainly against GPIIb/IIIa and GPIa/IIa, were more frequently detectable in the ELISA than in the solid-phase assay, whereas weakly positive results have to be interpreted cautiously. ConclusionELISA, solid-phase assay, and LCT showed highly divergent results. Due to several limitations, the additional analysis by the monoclonal antibody-specific immobilization of platelet antigen (MAIPA)assay is highly recommended.

Searchable abstracts of presentations at key conferences on calcified tissues ISSN 2052-1219 (online)

Abstract Abstract 2189 Heparin-induced thrombocytopenia (HIT), caused by antibodies against heparin/platelet factor 4 (HPF4) complex, is a rare but potentially serious side effect of heparin therapy where due to high mortality, rapid diagnosis is crucial. For the detection of HPF4 antibodies we compared the new nanoparticle-based lateral-flow immunoassay (LFI-HIT, Milenia Biotec, Germany) and a particle gel immunoassay (PaGIA, BioRad, Germany) with an IgG-specific-PF4/polyanion enzyme-linked immunosorbent assay (IgG-ELISA, GTI Diagnostics, USA). Sera from 121 patients (54/67 f/m, median 73 years, range 14–94) with suspected HIT were prospectively tested. The LFI-HIT and the PaGIA were evaluated visually, the IgG-ELISA was positive at an optical density (OD) cutoff > 0.4. For most of the positive samples, the functional heparin-induced platelet activation (HIPA) assay was additionally performed to detect false positive serological results and to confirm a clinically relevant HIT by in vitro platelet-activation. Regarding HIT as a clinico-pathological syndrome, characteristics for HIT were evaluated for each patient by the 4Ts scoring system and divided into high, intermediate or low risk. Results of serological analyses and OD values are summarized in the table. Ten of 121 samples were positive in the LFI-HIT, 10/10 positive in the PaGIA and 8/10 positive in the IgG-ELISA. The HIPA was tested in 9/10 samples and was positive in 8/9 samples. Of the 2 samples positive for LFI-HIT and PaGIA but negative in the ELISA, 1 was HIPA positive, 1 HIPA negative, resulting in a specificity of 88.9% for the LFI-HIT assay correlated to the HIPA. From 111/121 LFI-HIT-negative samples, 2 were positive in the PaGIA, the IgG-ELISA (OD 1.318 and 2,019) and in the HIPA. Seven of the 111 LFI-HIT negative samples were positive only in the IgG-ELISA. Due to marginal positive reactions of 5/7 samples in the ELISA with OD values between 0.4 to 0.5, only 2 LIF-HIT negative IgG-ELISA positive samples were tested by HIPA and 1/2 was positive. Based on the ELISA, the sensitivity of the LFI-HIT was 91.9% (102/111 negative samples also negative in the ELISA) in contrast to 93.1% of the PaGIA. The specificity of the LFI-HIT was 80% (LFI-HIT and IgG-ELISA positive), compared to 57.9% of the PaGIA. Notably, the clinical risk estimated by the 4Ts score system (received from 92/121 patients) did not correlate with laboratory diagnosis of HIT, probably due to inadequate evaluation. Concluding our data, a reliable exclusion of HIT by rapid testing with the LFI-HIT only seems possible with additional analysis of HPF4 antibodies by IgG-ELISA and/or HIPA assay. LFI-HIT PaGIA IgG-ELISA OD IgG-ELISA HIPA assay Median (range) Samples n=121 Pos 10 Pos 10 Pos 8 2.366 (0.902-3.000) 7/7 pos Neg 2 0.199 and 0.170 1/2 pos, 1/2 neg Neg 0 - - - - Neg 111 Pos 9 Pos 2 1.318 and 2.019 2/2 pos Neg 7 0.110 (0.054-0.139) 6/6 neg Neg 102 Pos 7 0.436 (0.404-1.463) 1/2 pos, 1/2 neg Neg 95 0.082 (0.013-0.376) Disclosures: No relevant conflicts of interest to declare.

CONTEXT:Sclerostin is produced by osteocytes and inhibits bone formation through the Wnt/β-catenin-signaling pathway. Only limited data are available on circulating sclerostin levels in healthy subjects.OBJECTIVE:We aimed to evaluate the correlation between sclerostin and physical activity, anthropometric, and biochemical variables.DESIGN, SETTING, AND PARTICIPANTS:We conducted a cross-sectional observational study in 161 healthy adult men and premenopausal women aged 19 to 64 yr (mean age, 44 ± 10).INTERVENTION(S):There were no interventions.MAIN OUTCOME MEASURE(S):Serum sclerostin levels were associated with body composition, bone mineral density, physical activity, and various biochemical parameters.RESULTS:A positive correlation between age and sclerostin in both men (r = 0.37; P < 0.001) and premenopausal women (r = 0.66; P < 0.001) was found. Men had significantly higher sclerostin levels than women (49.8 ± 17.6 vs. 37.2 ± 15.2 pmol/liter; P < 0.001). However, after adjustment for age, bone mineral content (BMC), physical activity, body mass index (BMI), and renal function, sclerostin levels did not differ (P = 0.543). Partial correlation analysis adjusted for age, gender, and kidney function revealed a significant positive correlation between sclerostin levels and BMC, bone mineral density, BMI, and android/gynoid fat and a significant negative correlation with serum osteocalcin and calcium. The most physically active quartile had significantly lower sclerostin levels compared to the least active quartile in a univariate analysis.CONCLUSIONS:In healthy adults, sclerostin serum levels correlate positively with age, BMI, and BMC and negatively with osteocalcin and calcium. Further studies in larger populations are needed to confirm our findings and to better understand their clinical implications.

Although blood donation is generally safe, a variety of risks and complications exist, the most common being iron deficiency, vasovagal reactions and citrate-related events. In the last decades, extensive efforts have significantly improved recipient and product safety, but there is still great potential to optimise donor care. Many therapies in modern medicine depend on the prompt availability of blood products, therefore it is crucial to maintain a motivated and healthy donor pool in view of a limited number of healthy volunteers willing and able to give blood or blood components. We present a comprehensive review on adverse events addressing all types of blood donation including whole blood, plasma, platelet, peripheral blood stem cell, leucocyte and bone marrow donation. In addition, we outline strategies for the prevention and treatment of these events and give a blueprint for future research in this field.