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In this study, a range of miniaturized Ag/AgCl reference electrodes with various layouts were successfully fabricated on wafer-level silicon-based substrates with metallic intermediate layers by precisely controlling the electrochemical deposition of Ag, followed by electrochemical chlorination of the deposited Ag layer. The structure, as well as the chemical composition of the electrode, were characterized with SEM & EDS. The results showed that the chlorination is very sensitive to the applied electric field and background solution. Potentiostatic chlorination, in combination with an adjusted mushroom-shaped Ag sealing deposition, enabled the formation of electrochemical usable Ag/AgCl layers. The stability of the electrodes was tested using open circuit potential (OCP) measurement. The results showed that the reference electrodes stayed stable for 300 s under 3 M KCl solution. The first stage study showed that the stability of the Ag/AgCl reference electrode in a chip highly depends on chip size design, chlorination conditions, and a further protection layer.

Accurate measurements of actual evapotranspiration (ETa) play an important role in understanding land surface processes and agricultural management. Two of the most commonly used and established methods for quantifying ETa are eddy covariance (EC) and weighable lysimeters measurements. Previous studies on hourly or daily basis indicated sometimes large differences between the ETa of the two methods (Δly-EC). It is still unclear which factors influence these differences. Here, we examine and compare half-hourly ETa measurements from EC (ETEC) and weighable lysimeters (ETly) at four different sites. The four sites span a climatic gradient from humid conditions at a pre-alpine (Fendt, DE) and a mid-mountain grassland (Rollesbroich, DE) to semi-arid conditions at two sites with a natural grass and shrub (Els Plans, ES) and a tree-grass ecosystem (Majadas de Tiétar, ES). We used a boosted regression tree method to identify environmental drivers of Δly-EC during day and night at the half-hourly resolution.Our results revealed that substantial differences were found with a mean annual Δly-EC of 117 mm, and Δly-EC displayed obvious spatiotemporal variabilities across the sites. Energy balance non-closure of EC was found to be the most important factor contributing to the large annual Δly-EC, especially at Majadas de Tiétar site. With the distinct climatic gradient, Δly-EC was negatively correlated with mean annual wind speed and vapor pressure deficit when they reached a specific level. Monthly ETEC and ETly agreed well with Δly-EC peaking in summer at the sites in Germany, while Δly-EC peaked earlier due to the different climate in Spain. Differences in grass height caused by field management and EC footprint also affected Δly-EC, especially at daily timescales for the pre-alpine and the mid-mountain grassland ecosystem. The relative impacts of different environmental variables to half-hourly ETEC and ETly were almost the same with soil water content (SWC) being more important for nighttime ETly. Meanwhile, we found that the dominant controlling factors of daytime Δly-EC changed with climatic conditions, but nighttime Δly-EC were mainly regulated by SWC. These findings provide a critical evaluation for the roles of climatic and land surface conditions on turbulent flux dynamics from different measurements, which has important implications for ecosystem water and energy balance.

Deforestation has a wide range of effects on hydrological and geochemical processes. Dissolved organic carbon (DOC) dynamics, a sensitive environmental change indicator, is expected to be affected by deforestation, with changes in atmospheric sulfur (S) deposition compounding this. However, how precisely anthropogenic disturbance (deforestation) under a declining atmospheric S input scenario affects the underlying spatiotemporal dynamics and relationships of river DOC and sulfate with hydro-climatological variables e.g., stream water temperature, runoff, pH, total dissolved iron (Fetot), and calcium (Ca2+) remains unclear. We, therefore, examined this issue within the TERENO Wüstebach catchment (Eifel, Germany), where partial deforestation had taken place in 2013. Wavelet transform coherence (WTC) analysis was applied based on a 10-year time series (2010–2020) from three sampling stations, whose (sub) catchment areas have different proportions of deforested area (W10: 31%, W14: 25%, W17: 3%). We found that water temperature and DOC, sulfate, and Fetot concentrations showed distinct seasonal patterns, with DOC averaging concentrations ranging from 2.23 (W17) to 4.56 (W10) mg L-1 and sulfate concentration ranging from 8.04 (W10) to 10.58 (W17) mg L-1. After clear-cut, DOC significantly increased by 59, 58% in the mainstream (W10, W14), but only 26% in the reference stream. WTC results indicated that DOC was negatively correlated with runoff and sulfate, but positively correlated with temperature, Ca2+, and Fetot. The negative correlation between DOC with runoff and sulfate was apparent over the whole examined 10-year period in W17 but did end in W10 and W14 after the deforestation. Sulfate was highly correlated with stream water temperature, runoff, and Fetot in W10 and W14 and with a longer lag time than W17. Additionally, pH was stronger correlated (higher R2) with sulfate and DOC in W17 than in W10 and W14. In conclusion, WTC analysis indicates that within this low mountainous forest catchment deforestation levels over 25% (W10 and W14) affected the coupling of S and C cycling substantially more strongly than “natural” environmental changes as observed in W17.

AbstractIn this study, we describe a methodology to derive climate analogue cities for spatially highly resolved future climate scenarios. For the computation, a reduced and in hindsight bias-adjusted EURO-CORDEX EUR-11 dataset is used based on two climate scenarios (RCP4.5 and RCP8.5). A total of 389 European cities are processed by the algorithm, which uses five statistical climate variables (2-m air temperature average and amplitude, precipitation sum and amplitude, correlation between 2-m air temperature average and precipitation sum). Additionally, extreme weather events (hot days, summer days, tropical nights, extreme precipitation events) are calculated for further comparison and validation. Finding an appropriate analogue permits a more accurate derivation and depiction of necessary climate adaptation efforts and therefore assist decision-making in city planning. As an example of our method, we searched for plausible climate twins for the mid-sized city of Aachen (Germany) at the end of the twenty-first century. Our results show that the French city of Dijon is highly likely to become Aachen’s climate twin by the end of the century for RCP4.5. As for the scenario RCP8.5, no clear European analogue city could be determined, indicating that the city might enter a novel climate. The nearest match suggests the cities of Florence and Prato in Tuscany. However, considering climate indices, the encompassing region of the French–Spanish city triangle Bordeaux–Toulouse–Bilbao is a better fit. The developed algorithm can be applied to any of the cities included in the dataset.

Boundary-Layer Meteorology (2022). doi:10.1007/s10546-022-00700-1, Published by Springer, Dordrecht [u.a.]

Infection-neutralizing antibody responses after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection or coronavirus disease 2019 vaccination are an essential component of antiviral immunity. Antibody-mediated protection is challenged by the emergence of SARS-CoV-2 variants of concern (VoCs) with immune escape properties, such as omicron (B.1.1.529), which is rapidly spreading worldwide. Here we report neutralizing antibody dynamics in a longitudinal cohort of coronavirus disease 2019 convalescent and infection-naive individuals vaccinated with mRNA BNT162b2 by quantifying SARS-CoV-2 spike protein antibodies and determining their avidity and neutralization capacity in serum. Using live-virus neutralization assays, we show that a superior infection-neutralizing capacity against all VoCs, including omicron, developed after either two vaccinations in convalescents or a third vaccination or breakthrough infection of twice-vaccinated, naive individuals. These three consecutive spike antigen exposures resulted in an increasing neutralization capacity per anti-spike antibody unit and were paralleled by stepwise increases in antibody avidity. We conclude that an infection-plus-vaccination-induced hybrid immunity or a triple immunization can induce high-quality antibodies with superior neutralization capacity against VoCs, including omicron.

On November 26, 2021, the World Health Organization classified B.1.1.529 as a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variant of concern (VoC), named omicron. Spike-gene dropouts in conventional SARS-CoV-2 PCR systems have been reported over the last weeks as indirect diagnostic evidence for the identification of omicron. Here, we report the combination of PCRs specific for heavily mutated sites in the spike gene and nanopore-based full-length genome sequencing for the rapid and sensitive identification of the first four COVID-19 patients diagnosed in Germany to be infected with omicron on November 28, 2021. This study will assist the unambiguous laboratory-based diagnosis and global surveillance for this highly contagious VoC with an unprecedented degree of humoral immune escape. Moreover, we propose that specialized diagnostic laboratories should continuously update their assays for variant-specific PCRs in the spike gene of SARS-CoV-2 to readily detect and diagnose emerging variants of interest and VoCs. The combination with established nanopore sequencing procedures allows both the rapid confirmation by whole genome sequencing as well as the sensitive identification of newly emerging variants of this pandemic β-coronavirus in years to come.

Figure S1 Recurrently mutated genes and comparison to TCGA cohort. Figure S2 Stabililty of recurrently mutated genes over disease course. Figure S3 Variant allele frequency plots from diagnosis to relapse for each individual patient. Figure S4 Mutation patterns of individual genes. Lines represent mutations in individual patients. Figure S5 Detection of the pre-existence of gained variants at initial diagnosis. Figure S6 Mutations in AML-associated functional pathways. Figure S7 Deletion of exons 3-10 of the KDM6A gene in the MM-6 cell line. The sister cell line MM-1 is not affected. Deletions were detected by quantitative MLPA analysis and the peak ratio for each KDM6A exon specific probe is shown. Graph represents the results from two independent experiments. Figure S8 H3K27 methylation in the AML cell lines MM-1 and MM-6. The global mono-, di- and tri-methylation of H3K27 in MM-1 and MM-6 was analyzed by Western blot and normalized to H3. The median of three independent experiments is shown. P-Values were calculated using a two-tailed, unpaired Student's t-test. Figure S9 The MM-6 cell line is resistant to cytarabine (Ara-C) but not to Daunorubicin or AC220. Error bars indicate mean {plus minus} s.d. of at least three independent experiments. **P75th percentile) with clinical outcome according to gender in the AMLCG-99 trial (NCT00266136). Figure S11 Proportion of transversions from diagnosis to relapse. Transversion frequencies are shown for lost (blue), stable (orange) and gained (red) mutations. Dots represent individual patients. Figure S12 Median coverage in targeted sequencing of persistent and non-persistent DNMT3A variant positions at complete remission (CR). Figure S13 Clinical outcome according to mutation persistence at remission.

Detailed patient characteristics

Purpose: To study mechanisms of therapy resistance and disease progression, we analyzed the evolution of cytogenetically normal acute myeloid leukemia (CN-AML) based on somatic alterations.Experimental Design: We performed exome sequencing of matched diagnosis, remission, and relapse samples from 50 CN-AML patients treated with intensive chemotherapy. Mutation patterns were correlated with clinical parameters.Results: Evolutionary patterns correlated with clinical outcome. Gain of mutations was associated with late relapse. Alterations of epigenetic regulators were frequently gained at relapse with recurring alterations of KDM6A constituting a mechanism of cytarabine resistance. Low KDM6A expression correlated with adverse clinical outcome, particularly in male patients. At complete remission, persistent mutations representing preleukemic lesions were observed in 48% of patients. The persistence of DNMT3A mutations correlated with shorter time to relapse.Conclusions: Chemotherapy resistance might be acquired through gain of mutations. Insights into the evolution during therapy and disease progression lay the foundation for tailored approaches to treat or prevent relapse of CN-AML. Clin Cancer Res; 24(7); 1716–26. ©2018 AACR.

JGR / Biogeosciences 128(2), e2022JG007197 (2023). doi:10.1029/2022JG007197, Published by Wiley, Hoboken, NJ

Although it has been shown that climate warming has steadily increased the length of the growing season (LGS) in Europe, we present new evidence that this trend reversed during last decade. Warmer European winter and spring weather combined with adequate soil moisture still results in early greening, albeit at slower rates than in the past. However, the recent (2014-2020) accelerated shift toward earlier onset of dormancy has resulted in a shortening of LGS compared to previous years. The results show that this is mainly due to higher atmospheric water demand (AWD) in summer. The higher AWD stresses the vegetation even though there is still enough water, but the vegetation cannot provide the needed water for transpiration because the water transport system is inadequate, or the root system is adapted to conditions other than the current condition. Our results have implications for future management of European ecosystems in a warmer world.

During 2022, the COVID-19 pandemic has been dominated by the variant of concern (VoC) Omicron (B.1.1.529) and its rapidly emerging subvariants, including Omicron-BA.1 and -BA.2. Rapid antigen tests (RATs) are part of national testing strategies to identify SARS-CoV-2 infections on site in a community setting or to support layman’s diagnostics at home. We and others have recently demonstrated an impaired RAT detection of infections caused by Omicron-BA.1 compared to Delta. Here, we evaluated the performance of five SARS-CoV-2 RATs in a single-centre laboratory study examining a total of 140 SARS-CoV-2 PCR-positive respiratory swab samples, 70 Omicron-BA.1 and 70 Omicron-BA.2, as well as 52 SARS-CoV-2 PCR-negative swabs collected from March 8th until April 10th, 2022. One test did not meet minimal criteria for specificity. In an assessment of the analytical sensitivity in clinical specimen, the 50% limit of detection (LoD50) ranged from 4.2 × 104 to 9.2 × 105 RNA copies subjected to the RAT for Omicron-BA.1 compared to 1.3 × 105 to 1.5 × 106 for Omicron-BA.2. Overall, intra-assay differences for the detection of Omicron-BA.1-containing and Omicron-BA.2-containing samples were non-significant, while a marked overall heterogeneity among the five RATs was observed. To score positive in these point-of-care tests, up to 22-fold (LoD50) or 68-fold (LoD95) higher viral loads were required for the worst performing compared to the best performing RAT. The rates of true-positive test results for these Omicron subvariant-containing samples in the highest viral load category (Ct values

Sarcopenic dysphagia (SD) is swallowing difficulty associated with loss of generalized skeletal muscles and swallowing-related muscles. Diagnostic criteria for SD were suggested, yet there is a variability in instruments and cut-offs used. The aim of the current review is to critically evaluate tools used in diagnosis of sarcopenic dysphagia in the elderly. Comprehensive review of the literature was performed. Studies were qualitatively evaluated for the diagnostic tools used to make a diagnosis of "sarcopenic dysphagia" and compared to the known diagnostic criteria for SD and other accepted measures. Fourteen studies (N = 10,282) were selected from a search yield of 331 de-duplicated studies. Ninety-three percent of studies (13/14) were conducted in Japan. All subjects included were over the age of 65 years old (mean, 76.5 years). Various tools were used to assess sarcopenia including handgrip strength (14/14 of studies), followed by skeletal muscle mass/index (7/14), tongue pressure, gait speed, and calf circumference in 5/14 studies. The most commonly tool used for dysphagia and/or swallowing dysfunction was the food level intake scale (5/14 of studies) followed by the functional oral intake scale (3/14). The 100-mL water swallow test was used in 2 of the 14 included SD studies. Fiberoptic endoscopic evaluation of swallowing, videofluoroscopic swallowing study, EAT-10 questionnaire, and standardized swallow assessment were each used in only one SD study. Further research is required to validate SD diagnostic tools, establish cut-offs in different populations, and investigate their role in screening of dysphagia and swallowing dysfunction in the elderly.

Sulfur deficiency-induced proteins SDI1 and SDI2 play a fundamental role in sulfur homeostasis under sulfate-deprived conditions (−S) by downregulating glucosinolates. Here, we identified that besides glucosinolate regulation under –S, SDI1 downregulates another sulfur pool, the S-rich 2S seed storage proteins in Arabidopsis (Arabidopsis thaliana) seeds. We identified that MYB28 directly regulates 2S seed storage proteins by binding to the At2S4 promoter. We also showed that SDI1 downregulates 2S seed storage proteins by forming a ternary protein complex with MYB28 and MYC2, another transcription factor involved in the regulation of seed storage proteins. These findings have significant implications for the understanding of plant responses to sulfur deficiency., Sulfur Deficiency Induced1 (SDI1) upregulates S-poor seed storage proteins in favor of S-rich seed storage proteins.

Coccolithophores are globally abundant, calcifying microalgae that have profound effects on marine biogeochemical cycles, the climate, and life in the oceans. They are characterized by a cell wall of CaCO3 scales called coccoliths, which may contribute to their ecological success. The intricate morphologies of coccoliths are of interest for biomimetic materials synthesis. Despite the global impact of coccolithophore calcification, we know little about the molecular machinery underpinning coccolithophore biology. Working in the model Emiliania huxleyi, a globally distributed bloom-former, we deploy a range of proteomic strategies to identify coccolithogenesis-related proteins. These analyses are underpinned by a new genome, with gene models derived from long-read transcriptome sequencing, which revealed many novel proteins specific to the calcifying haptophytes. Our experiments provide fresh insights into proteins involved in various aspects of coccolithogenesis. Our improved genome, complemented with transcriptomic and proteomic data, constitute an excellent resource for investigating fundamental aspects of coccolithophore biology.

Identification of fusion genes in clinical routine is mostly based on cytogenetics and targeted molecular genetics, such as metaphase karyotyping, fluorescence in situ hybridization and reverse-transcriptase polymerase chain reaction. However, sequencing technologies are becoming more important in clinical routine as processing time and costs per sample decrease. To evaluate the performance of fusion gene detection by RNAsequencing compared to standard diagnostic techniques, we analyzed 806 RNA-sequencing samples from patients with acute myeloid leukemia using two state-of-the-art software tools, namely Arriba and FusionCatcher. RNA-sequencing detected 90% of fusion events that were reported by routine with high evidence, while samples in which RNA-sequencing failed to detect fusion genes had overall lower and inhomogeneous sequence coverage. Based on properties of known and unknown fusion events, we developed a workflow with integrated filtering strategies for the identification of robust fusion gene candidates by RNA-sequencing. Thereby, we detected known recurrent fusion events in 26 cases that were not reported by routine and found discrepancies in evidence for known fusion events between routine and RNA-sequencing in three cases. Moreover, we identified 157 fusion genes as novel robust candidates and comparison to entries from ChimerDB or Mitelman Database showed novel recurrence of fusion genes in 14 cases. Finally, we detected the novel recurrent fusion gene NRIP1- MIR99AHG resulting from inv(21)(q11.2;q21.1) in nine patients (1.1%) and LTN1-MX1 resulting from inv(21)(q21.3;q22.3) in two patients (0.25%). We demonstrated that NRIP1-MIR99AHG results in overexpression of the 3' region of MIR99AHG and the disruption of the tricistronic miRNA cluster miR-99a/let-7c/miR-125b-2. Interestingly, upregulation of MIR99AHG and deregulation of the miRNA cluster, residing in the MIR99AHG locus, are known mechanisms of leukemogenesis in acute megakaryoblastic leukemia. Our findings demonstrate that RNA-sequencing has a strong potential to improve the systematic detection of fusion genes in clinical applications and provides a valuable tool for fusion discovery.

Authorities regulate online transactions to protect consumers from unintended negative consequences. Regulatory cues (e.g. certifications) enable consumers to identify formally approved online gamb...

ZusammenfassungDie Covid-Pandemie verstärkt globale Entwicklungen, die bereits zuvor ihre Schatten voraus warfen. Eine neue Ära der Großmachtkonflikte bricht an, die Demokratie gerät weltweit in die Defensive. Insbesondere der Aufstieg Chinas fordert die Staatengemeinschaft heraus. Internationale Organisationen stehen unter Druck, während Herausforderungen wie der Klimawandel multilaterale Antworten erfordern. Deutschland wird daher international mehr Verantwortung übernehmen müssen. Der Beitrag stellt vor diesem Hintergrund die außen- und sicherheitspolitischen Positionen der Fraktion der Freien Demokraten vor.

Background High infection rates among healthcare personnel in an uncontained pandemic can paralyze health systems due to staff shortages. Risk constellations and rates of seroconversion for healthcare workers (HCWs) during the first wave of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic are still largely unclear. Methods Healthcare personnel (n = 300) on different organizational units in the LMU Munich University Hospital were included and followed in this prospective longitudinal study from 24 March until 7 July 2020. Participants were monitored in intervals of 2 to 6 weeks using different antibody assays for serological testing and questionnaires to evaluate risk contacts. In a subgroup of infected participants, we obtained nasopharyngeal swabs to perform whole-genome sequencing for outbreak characterization. Results HCWs involved in patient care on dedicated coronavirus disease 2019 (COVID-19) wards or on regular non–COVID-19 wards showed a higher rate of SARS-CoV-2 seroconversion than staff in the emergency department and non-frontline personnel. The landscape of risk contacts in these units was dynamic, with a decrease in unprotected risk contacts in the emergency department and an increase on non–COVID-19 wards. Both intensity and number of risk contacts were associated with higher rates of seroconversion. On regular wards, staff infections tended to occur in clusters, while infections on COVID-19 wards were less frequent and apparently independent of each other. Conclusions Risk of SARS-CoV-2 infection for frontline HCWs was increased during the first pandemic wave in southern Germany. Stringent measures for infection control are essential to protect all patient-facing staff during the ongoing pandemic.