Impact of the COVID-19 outbreak on blood supply in two large university hospitals.

OBJECTIVE: This study aimed to examine the relationship between the decrease in elective procedures and the need for blood donation during the novel coronavirus disease (COVID-19) pandemic at university hospitals. BACKGROUND: The COVID-19 pandemic has immensely impacted transfusion medicine. By cancelling elective surgery, the German government hoped to increase the available resources for patients infected with COVID-19, especially in intensive care units, and prevent the shortage of blood products. METHODS/MATERIALS: Over 26 weeks, from the 3rd of February 2020 to the 2nd of August 2020, during the first phase of the pandemic, we assessed the number of crossmatches, blood group typing, use of donated blood, and case mix indices by retrospectively analysing data from two major university hospitals' information systems in Essen and Hamburg, Germany. Data were pooled, analysed, and compared with that of the same period in the previous year. RESULTS: Following the cessation of elective procedures, the number of requests for crossmatches and blood group typing significantly decreased in 2020 compared to that in 2019. However, the number of blood transfusions required was reduced to a lesser extent. The number of outpatient and inpatient cases significantly decreased, whereas the cases requiring transfusion decreased only. CONCLUSION: During the initial phase of the pandemic, transfusion medicine, especially in large institutions, faced an almost unchanged high demand for donated blood. This should be considered regarding personnel and blood donation allocations. Therefore, we developed a monitoring system to display the availability of blood products in real-time. The quick and easy display of in-stock and expiring blood products can optimise the use of this valuable resource.

Impaired Degradation of Neutrophil Extracellular Traps: A Possible Severity Factor of Elderly Male COVID-19 Patients.

Neutrophil extracellular traps (NETs) have been described as a potential trigger of severe COVID-19. NETs are known as extracellular DNA fibers released by neutrophils in response to infection. If the host is unable to balance efficient clearance of NETs by dornases (DNases), detrimental consequences occur. Elevated levels of NETs in COVID-19 patients are associated with higher risk of morbid thrombotic complications. Here, we studied the level of NET markers and DNase activity in a cohort of COVID-19 patients compared to healthy controls. Our data confirmed an increased level of NET markers in the plasma of COVID-19 patients, with a higher level in male compared to female patients. At the same time, there was an increased DNase activity detectable in COVID-19 patients compared to healthy controls. Importantly, there was a negative correlation of DNase activity with the age of male patients. The antimicrobial peptide LL-37, which is known to stabilize NETs against DNase degradation, is embedded in NETs upon severe acute respiratory syndrome coronavirus-2-infection. The LL-37 plasma level correlates with the NET-marker level in male COVID-19 patients, indicating a potential role of LL-37 in the risk of NET-associated thrombosis in male COVID-19 patients by stabilizing NETs against DNase degradation. In conclusion, our data identify two potential risk factors of elderly male patients which may lead to inefficient NET degradation and a subsequently higher risk of NET-associated thrombosis during COVID-19: reduced DNase activity and an increased LL-37 level.

High estradiol and low testosterone levels are associated with critical illness in male but not in female COVID-19 patients: a retrospective cohort study.

Male sex was repeatedly identified as a risk factor for death and intensive care admission. However, it is yet unclear whether sex hormones are associated with disease severity in COVID-19 patients. In this study, we analysed sex hormone levels (estradiol and testosterone) of male and female COVID-19 patients (n = 50) admitted to an intensive care unit (ICU) in comparison to control non-COVID-19 patients at the ICU (n = 42), non-COVID-19 patients with the most prevalent comorbidity (coronary heart diseases) present within the COVID-19 cohort (n = 39) and healthy individuals (n = 50). We detected significantly elevated estradiol levels in critically ill male COVID-19 patients compared to all control cohorts. Testosterone levels were significantly reduced in critically ill male COVID-19 patients compared to control cohorts. No statistically significant differences in sex hormone levels were detected in critically ill female COVID-19 patients, albeit similar trends towards elevated estradiol levels were observed. Linear regression analysis revealed that among a broad range of cytokines and chemokines analysed, IFN-γ levels are positively associated with estradiol levels in male and female COVID-19 patients. Furthermore, male COVID-19 patients with elevated estradiol levels were more likely to receive ECMO treatment. Thus, we herein identified that disturbance of sex hormone metabolism might present a hallmark in critically ill male COVID-19 patients.

Sustained Response After Remdesivir and Convalescent Plasma Therapy in a B-Cell-Depleted Patient With Protracted Coronavirus Disease 2019 (COVID-19).

We provide detailed clinical, virological, and immunological data of a B-cell-depleted patient treated with obinutuzumab for follicular lymphoma with protracted coronavirus disease 2019 (COVID-19) and viremia. A sustained response was achieved after 2 courses of remdesivir and subsequent convalescent plasma therapy. Immunocompromised patients might require combined and prolonged antiviral treatment regimens.

Clinical evaluation of five different automated SARS-CoV-2 serology assays in a cohort of hospitalized COVID-19 patients.

BACKGROUND: The global market for SARS-CoV-2-immunoassays is becoming ever more crowded with antibody-tests of various formats, targets and technologies, careful evaluation is crucial for understanding the implications of individual test results. Here, we evaluate the clinical performance of five automated immunoassays on a set of clinical samples. METHODS: Serum/plasma samples of 75 confirmed COVID-19 patients and 320 pre-pandemic serum samples of healthy blood donors were subjected to two IgG and three total antibody SARS-CoV-2-immunoassays. All test setups were automated workflows. RESULTS: Positivity of assays (onset of symptoms > 10 days) ranged between 68.4 % and 81.6 % (Diasorin 68.4 %, Euroimmun 70.3 %, Siemens 73.7 %, Roche 79.0 % and Wantai 81.6 %). All examined assays demonstrated high specificity of >99 % (Euroimmun, Diasorin: 99.1 %, Wantai: 99.4 %) but only two reached levels above 99.5 % (Roche: 99.7 %, Siemens 100 %). Interestingly, there was no overlap in false positive results between the assays. The strongest correlation of quantitative results was observed between the Diasorin and Euroimmun IgG tests (r2 = 0.76). Overall, we observed no difference in the distribution of test results between female and male patients (p-values: 0.18-0.87). A significant difference between severely versus critically ill patients was demonstrated for the Euroimmun, Diasorin, Wantai and Siemens assays (p-values:0.041). CONCLUSION: All assays showed good clinical performance. Our data confirm that orthogonal test strategies as recommended by the CDC can enhance clinical specificity. However, the suboptimal rates of test positivity found at time of hospitalization in this cohort underline the importance of molecular diagnostics to rule out/confirm active infection with SARS-CoV-2.

Blood donation and donors: insights from a large German teaching hospital (2008-2017).

BACKGROUND AND OBJECTIVES: The availability of blood and blood products is crucial for the provision of high-quality hospital services. We analyse changes in whole blood donations, donors and their behaviour over 9 years at a large German teaching hospital. MATERIALS AND METHODS: A descriptive analysis using data from over 34 000 donors and 265 000 donations from a large university hospital's blood centre was conducted using data from July 2008 to December 2017. The analysis focussed on (a) whole blood donations and (b) donor characteristics and how they changed over time. We categorized donors into four categories according to their donation activity (First-Time, Highly Active, Active and Reactivated). RESULTS: We observed falling donations over time and that donors donated less frequently. Consequently, we show a downward trend in the number of Highly Active donors, whilst First-Time donors remained stable. We also provide evidence that donors donated well below their capacity and that the blood type of donors appeared to be in line with the wider German donor population. Lastly, we show a sharp drop in the return rates of First-Time donors over time. CONCLUSION: We recommend that Highly Active donors and former Highly Active donors are more carefully considered when planning donor engagement strategies and effort made in (at the very least) maintaining their donation activity. Our results in the context of the literature highlight the need for further research into the changing attitudes towards blood donation and prosocial activities.

Course of HEV viremia and anti-HEV IgM/IgG response in asymptomatic blood donors.

BACKGROUND: Globally, an estimated 20 million Hepatitis E infections occur every year. The course of viremia and antibody response has been investigated in patients with symptomatic hepatitis E. However, the majority of HEV infections in industrialized countries take a subclinical course. OBJECTIVES: To investigate the course of HEV viremia and epitope specific anti-HEV IgM/IgG response in asymptomatic blood donors in order to understand the immune response and viral clearance in asymptomatic blood donors with HEV infections. METHODS: In this study 27 HEV viremic donors were identified by HEV-PCR during routine screening of blood donors and the course of anti-HEV IgM/IgG and HEV-RNA was retrospectively studied using RT-PCR and a commercial immunoblot (Mikrogen®) allowing classification of the antibody response according to HEV epitopes. RESULTS: At time of donation, serological testing failed to identify viremic donors as 70.4% had no detectable antibody response. Anti-HEV IgM could be detected in 22.2% of viremic donors while anti-HEV IgG could be found in 7.4%. At least three donors experienced prolonged viremia beyond 100 days. Spontaneous HEV-RNA clearance within a median time span of 57 days was observed in all 27 donors. In all donors anti-HEV IgG specific for the immunogenic viral epitope O2C could be detected in close temporal association with viral clearance. CONCLUSION: Serological testing is inappropriate for identifying HEV-viremic blood donors. Acute HEV infection in asymptomatic blood donors can persist for more than 100 days. HEV-RNA clearance coincided with the appearance of anti-HEV IgM/IgG confirming the importance of a B-cell mediated response in clearing acute infections. Anti-HEV IgM and IgG specific for the epitope O2C are associated with the clearance of HEV-viremia.

HEV-positive blood donations represent a relevant infection risk for immunosuppressed recipients.

BACKGROUND & AIMS: Routine HEV testing of blood products has recently been implemented in Great Britain and the Netherlands. The relevance of transfusion-transmitted HEV infections is still controversially discussed in Europe. METHODS: All blood donations at the University Medical Center Hamburg-Eppendorf were prospectively tested for HEV RNA by pooled PCR from October 2016 to May 2017. Reactive samples were individually retested. Additionally, stored samples from previous donations of positive donors were tested to determine the duration of HEV viraemia. HEV RNA-positive donors and a control cohort were asked to answer a questionnaire. RESULTS: Twenty-three out of 18,737 HEV RNA-positive donors were identified (0.12%). Only two of the positive donors (8.7%) presented with elevated aminotransferases at time of donation (alanine aminotransferase: 192 and 101 U/L). The retrospective analysis of all positive donors revealed that four asymptomatic donors had been HEV viraemic for up to three months with the longest duration of HEV viraemia exceeding four months. Despite the HEV-testing efforts, 14 HEV RNA-positive blood products were transfused into 12 immunocompromised and two immunocompetent patients. One recipient of these products developed fatal acute-on-chronic liver failure complicated by Pseudomonas septicemia. The questionnaire revealed that HEV RNA-positive donors significantly more often consumed raw pork meat (12 out of 18; 67%) than controls (89 out of 256; 35%; p = 0.01). In two donors, undercooked pork liver dishes were identified as the source of infection. HEV genotyping was possible in 7 out of 23 of HEV viraemic donors and six out of seven isolates belonged to HEV Genotype 3, Group 2. CONCLUSIONS: Prolonged HEV viraemia can be detected at a relatively high rate in Northern German blood donors, leading to transfusion-transmitted HEV infections in several patients with the risk of severe and fatal complications. Eating raw pork tartare represented a relevant risk for the acquisition of HEV infection. LAY SUMMARY: The relevance of transfusion-transmitted hepatitis E virus infections has been discussed controversially. Herein, we present the first report on routine hepatitis E virus screening of blood donations at a tertiary care centre in Germany. Hepatitis E viraemia was found at a relatively high rate of 0.12% among blood donors, which represents a relevant transfusion-related risk for vulnerable patient populations.

BLOOM: BLoom filter based oblivious outsourced matchings.

BACKGROUND: Whole genome sequencing has become fast, accurate, and cheap, paving the way towards the large-scale collection and processing of human genome data. Unfortunately, this dawning genome era does not only promise tremendous advances in biomedical research but also causes unprecedented privacy risks for the many. Handling storage and processing of large genome datasets through cloud services greatly aggravates these concerns. Current research efforts thus investigate the use of strong cryptographic methods and protocols to implement privacy-preserving genomic computations. METHODS: We propose FHE-BLOOM and PHE-BLOOM, two efficient approaches for genetic disease testing using homomorphically encrypted Bloom filters. Both approaches allow the data owner to securely outsource storage and computation to an untrusted cloud. FHE-BLOOM is fully secure in the semi-honest model while PHE-BLOOM slightly relaxes security guarantees in a trade-off for highly improved performance. RESULTS: We implement and evaluate both approaches on a large dataset of up to 50 patient genomes each with up to 1000000 variations (single nucleotide polymorphisms). For both implementations, overheads scale linearly in the number of patients and variations, while PHE-BLOOM is faster by at least three orders of magnitude. For example, testing disease susceptibility of 50 patients with 100000 variations requires only a total of 308.31 s (σ=8.73 s) with our first approach and a mere 0.07 s (σ=0.00 s) with the second. We additionally discuss security guarantees of both approaches and their limitations as well as possible extensions towards more complex query types, e.g., fuzzy or range queries. CONCLUSIONS: Both approaches handle practical problem sizes efficiently and are easily parallelized to scale with the elastic resources available in the cloud. The fully homomorphic scheme, FHE-BLOOM, realizes a comprehensive outsourcing to the cloud, while the partially homomorphic scheme, PHE-BLOOM, trades a slight relaxation of security guarantees against performance improvements by at least three orders of magnitude.

Coding of Tissue and Cell Preparations Using Eurocode.

Traceability of products requires their unique identification. In Germany blood products have been encoded by Eurocode since 1998. EU Directives 2004/23/EC, 2006/86/EC and 2015/565/EC demanded unique identification and safe traceability procedure also for tissues and cells. Eurocode IBLS e.V. and the German Society of Transfusion Medicine and Immunohematology (DGTI) working parties '!' within the Single European Code (SEC). Several data elements of Eurocode can be used to create the complete SEC data structure, except the tissue establishment number. This can be found on the EU Coding Platform in the internet. Consequently, existing software and labeling solutions in Eurocode format could be easily upgraded with SEC.

AB0/Rhesus Blood Group Does Not Influence Clinicopathological Tumor Characteristics or Oncological Outcome in Patients Undergoing Radical Prostatectomy.

OBJECTIVES: AB0 blood group is an inherited characteristic that has been associated with the incidence as well as the prognosis of several malignancies. The aim of the current study was to clarify the role of the blood group in cancer epidemiology and clinical outcome of patients with prostate cancer (PCa). METHODS: Data from 3,574 patients undergoing radical prostatectomy between 2009 and 2010 at a single European institution were retrospectively analyzed. The correlation of AB0 and Rhesus blood group with PCa-related characteristics and oncological outcome were evaluated using univariable and multivariable Cox proportional hazard models. RESULTS: Median follow-up was 36.9 months. The overall distributions of AB0, as well as Rhesus blood groups among patients with PCa, did not differ from the distribution observed in the normal population. There was no significant association between AB0/Rhesus blood groups and Gleason score, prostate volume, surgical margin, pT-stage, pN-status, or preoperative prostate-specific antigen level. In multivariable Cox regression analysis, no statistically significant correlation between AB0/Rhesus group and biochemical recurrence was observed (all p > 0.05). CONCLUSION: Our data suggest no relevant association of AB0/Rhesus blood group with adverse clinicopathological tumor characteristics or oncological outcome after surgery in contrast to several other malignancies.

ABO blood group IgM isoagglutinins interact with tumor-associated O-glycan structures in pancreatic cancer.

PURPOSE: The ABO gene locus is associated with the risk of developing pancreatic ductal adenocarcinoma (PDAC) resulting in an increased incidence in individuals with non-O blood groups. Up to 90% of PDAC specimens display alterations in mucin type O-GalNAc glycosylation. Because aberrant O-GalNAc glycans (Tn and T antigen) are structurally related to blood group A and B glycans, we investigated the role of IgM isoagglutinins in PDAC. EXPERIMENTAL DESIGN: Binding studies of IgM isoagglutinins toward blood group A, B, Tn antigen, and T antigen glycoconjugates from patients with PDAC and healthy individuals were conducted. Isoagglutinin titers and total IgM were compared between patients with PDAC and control group. An anti-A antibody was used for immunoprecipitation of aberrant O-glycosylated tumor proteins and subsequent mass spectromic analysis. RESULTS: We found that IgM isoagglutinins bind blood group antigens, Tn and T glycoconjugates as well as tumor-derived glycoproteins. Blood group A isoagglutinins exhibited a strong binding toward blood group B antigen and T antigen, whereas blood group B showed binding to blood group A antigen and Tn antigen. Furthermore, we confirmed a decreased frequency in individuals with blood group O and observed a significant decrease of IgM isoagglutinin titers in PDAC sera compared with control sera, whereas total IgM levels were unaltered. We identified new PDAC-derived O-GalNAc glycoproteins by mass spectrometry using a blood group A-specific antibody. CONCLUSION: Our data elucidated a novel interaction of blood group IgM isoagglutinins and PDAC O-GalNAc glycoproteins that may contribute to the pathogenesis and progression of pancreatic cancer.

Antibody prevalence to the 2009 pandemic influenza A (H1N1) virus in Germany: geographically variable immunity in winter 2010/2011.

Location- and age-specific prevalence of antibodies against 2009 pandemic influenza A (H1N1) virus were determined in sera of blood donors collected during winter 2010/2011 in Germany. Prevalence of antibodies at protective titres (HI ≥1:40) varied significantly between cities (24.13-83.67 %) throughout all age groups. However, high level antibodies (HI >1:80) were most prevalent among young individuals (18-29 and 30-39 years). Overall, this study demonstrates that older people (50-59 and 60-70 years) are no longer more likely to present protective antibody titres against 2009 pandemic influenza A (H1N1) virus than younger individuals. Furthermore, our data show a highly variable immunity among the German population in different major cities almost 2 years after the detection of first cases in Germany.