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Abstract Despite decades of research, the influence of climate on the export of dissolved organic carbon (DOC) from soil remains poorly constrained, adding uncertainty to global carbon models. The limited temporal range of contemporary monitoring data, ongoing climate reorganisation and confounding anthropogenic activities muddy the waters further. Here, we reconstruct DOC leaching over the last ~14,000 years using alpine environmental archives (two speleothems and one lake sediment core) across 4° of latitude from Te Waipounamu/South Island of Aotearoa New Zealand. We selected broadly comparable palaeoenvironmental archives in mountainous catchments, free of anthropogenically-induced landscape changes prior to ~1200 C.E. We show that warmer temperatures resulted in increased allochthonous DOC export through the Holocene, most notably during the Holocene Climatic Optimum (HCO), which was some 1.5–2.5 °C warmer than the late pre-industrial period—then decreased during the cooler mid-Holocene. We propose that temperature exerted the key control on the observed doubling to tripling of soil DOC export during the HCO, presumably via temperature-mediated changes in vegetative soil C inputs and microbial degradation rates. Future warming may accelerate DOC export from mountainous catchments, with implications for the global carbon cycle and water quality.

Abstract Recently, a novel cyclo-heptapeptide composed of alternating d,l-amino acids and a unique thiazolidine heterocycle, called lugdunin, was discovered, which is produced by the nasal and skin commensal Staphylococcus lugdunensis. Lugdunin displays potent antimicrobial activity against a broad spectrum of Gram-positive bacteria, including challenging-to-treat methicillin-resistant Staphylococcus aureus (MRSA). Lugdunin specifically inhibits target bacteria by dissipating their membrane potential. However, the precise mode of action of this new class of fibupeptides remains largely elusive. Here, we disclose the mechanism by which lugdunin rapidly destabilizes the bacterial membrane potential using an in vitro approach. The peptide strongly partitions into lipid compositions resembling Gram-positive bacterial membranes but less in those harboring the eukaryotic membrane component cholesterol. Upon insertion, lugdunin forms hydrogen-bonded antiparallel β-sheets by the formation of peptide nanotubes, as demonstrated by ATR-FTIR spectroscopy and molecular dynamics simulations. These hydrophilic nanotubes filled with a water wire facilitate not only the translocation of protons but also of monovalent cations as demonstrated by voltage-clamp experiments on black lipid membranes. Collectively, our results provide evidence that the natural fibupeptide lugdunin acts as a peptidic channel that is spontaneously formed by an intricate stacking mechanism, leading to the dissipation of a bacterial cell’s membrane potential.

Introduction Diabetes is a significant metabolic disorder characterized by high blood glucose levels, leading to various complications if not properly managed. Understanding these factors is crucial for early identification, effective prevention, and management of diabetes within this community. Our study focused on the prevalence and risk factors of diabetes in Puzhakkatiri, a village in Malappuram district, Kerala, aiming to provide insights into local epidemiological patterns and targeted public health interventions. Methodology The community-based cross-sectional study included 582 participants (?18 years) selected via multistage sampling from Puzhakkattiri Panchayath, Northern Kerala, India. Selected participants were invited to a camp that was held at the Government school in collaboration with the panchayath office, where participants had their BMI, waist-to-hip ratio, fasting blood sugar (FBS), and postprandial blood sugar (PPBS) levels measured. Sociodemographic data were collected through interviews, and the data were analyzed using SPSS software, with a p-value of 0.05 or lower indicating statistical significance. Results The study, with participants having a mean age of 39.13 ± 12.56 years, found a diabetes prevalence of 30% and an impaired glucose tolerance prevalence of 47.4%. Higher diabetes rates were observed among individuals over 40 years (37.2%), those divorced/widowed/separated (46.5%), those with lower education levels (40%), those in unskilled occupations (34.4%), and those with overweight BMI (33%) or high waist-to-hip ratios (34.5%). Significant associations (p-value

Abstract El Niño–Southern Oscillation dynamics affect global weather patterns, with regionally diverse hydrological responses posing critical societal challenges. The lack of seasonally resolved hydrological proxy reconstructions beyond the observational era limits our understanding of boundary conditions that drive and/or adjust El Niño–Southern Oscillation variability. Detailed reconstructions of past El Niño–Southern Oscillation dynamics can help modelling efforts, highlight impacts on disparate ecosystems and link to extreme events that affect populations from the tropics to high latitudes. Here, mid‐Holocene El Niño–Southern Oscillation and hydrological changes are reconstructed in the south‐west Pacific using a stalagmite from Niue Island, which represents the period 6.4–5.4 ka BP. Stable oxygen and carbon isotope ratios, trace elements and greyscale data from a U/Th‐dated and layer counted stalagmite profile are combined to infer changes in local hydrology at sub‐annual to multi‐decadal timescales. Principal component analysis reveals seasonal‐scale hydrological changes expressed as variations in stalagmite growth patterns and geochemical characteristics. Higher levels of host rock‐derived elements (Sr/Ca and U/Ca) and higher δ18O and δ13C values are observed in dark, dense calcite laminae deposited during the dry season, whereas during the wet season, higher concentrations of soil‐derived elements (Zn/Ca and Mn/Ca) and lower δ18O and δ13C values are recorded in pale, porous calcite laminae. The multi‐proxy record from Niue shows seasonal cycles associated with hydrological changes controlled by the positioning and strength of the South Pacific Convergence Zone. Wavelet analysis of the greyscale record reveals that El Niño–Southern Oscillation was continuously active during the mid‐Holocene, with two weaker intervals at 6–5.9 and 5.6–5.5 ka BP. El Niño–Southern Oscillation especially affects dry season rainfall dynamics, with increased cyclone activity that reduces hydrological seasonality during El Niño years.

Purpose: Stereotactic body radiotherapy (SBRT) is an effective treatment for adrenal gland metastases, but it is technically challenging and there are concerns about toxicity. We performed a multi-institutional pooled retrospective analysis to study clinical outcomes and toxicities after MR-guided SBRT (MRgSBRT) using for adrenal gland metastases. Methods and Materials: Clinical and dosimetric data of patients treated with MRgSBRT on a 0.35 T MR-Linac at 11 institutions between 2016 and 2022 were analyzed. Local control (LC), local progression-free survival (LPFS), distant progression-free survival (DPFS) and overall survival (OS) were estimated using Kaplan-Meier method and log-rank test. Results: A total of 255 patients (269 adrenal metastases) were included. Metastatic pattern was solitary in 25.9 % and oligometastatic in 58.0 % of patients. Median total dose was 45 Gy (range, 16–60 Gy) in a median of 5 fractions, and the median BED10 was 100 Gy (range, 37.5–132.0 Gy). Adaptation was done in 87.4 % of delivered fractions based on the individual clinicians' judgement. The 1- and 2- year LPFS rates were 94.0 % (95 % CI: 90.7–97.3 %) and 88.3 % (95 % CI: 82.4–94.2 %), respectively and only 2 patients (0.8 %) experienced grade 3 + toxicity. No local recurrences were observed after treatment to a total dose of BED10 > 100 Gy, with single fraction or fractional dose of > 10 Gy. Conclusions: This is a large retrospective multi-institutional study to evaluate the treatment outcomes and toxicities with MRgSBRT in over 250 patients, demonstrating the need for frequent adaptation in 87.4 % of delivered fractions to achieve a 1- year LPFS rate of 94 % and less than 1 % rate of grade 3 + toxicity. Outcomes analysis in 269 adrenal lesions revealed improved outcomes with delivery of a BED10 > 100 Gy, use of single fraction SBRT and with fraction doses > 10 Gy, providing benchmarks for future clinical trials.

[This corrects the article DOI: 10.1371/journal.pone.0254817.].

Abstract Secondary transports of patients from one hospital to another are indicated for medical reasons or to address local constraints in capacity. In particular, interhospital transports of critically ill infectious patients present a logistical challenge and can be key in the effective management of pandemic situations. The state of Saxony in Germany has two characteristics that allow for an extensive evaluation of secondary transports in the pandemic year 2020/2021. First, all secondary transports are centrally coordinated by a single institution. Second, Saxony had the highest SARS-CoV-2 infection rates and the highest COVID-19 associated mortality in Germany. This study evaluates secondary interhospital transports from March 2019 to February 2021 in Saxony with a detailed analysis of transport behaviour during the pandemic phase March 2020 to February 2021. Our analysis includes secondary transports of SARS-CoV-2 patients and compares them to secondary transports of non-infectious patients. In addition, our data show differences in demographics, SARS-CoV-2- incidences, ICU occupancy of COVID-19 patients, and COVID-19 associated mortality in all three regional health clusters in Saxony. In total, 12,282 secondary transports were analysed between March 1st, 2020 and February 28th, 2021, of which 632 were associated with SARS-CoV-2 (5.1%) The total number of secondary transports changed slightly during the study period March 2020 to February 2021. Transport capacities for non-infectious patients were reduced due to in-hospital and out-of-hospital measures and could be used for transport of SARS-CoV-2 patients. Infectious transfers lasted longer despite shorter distance, occurred more frequently on weekends and transported patients were older. Primary transport vehicles were emergency ambulances, transport ambulances and intensive care transport vehicles. Data analysis based on hospital structures showed that secondary transports in correlation to weekly case numbers depend on the hospital type. Maximum care hospitals and specialized hospitals show a maximum of infectious transports approximately 4 weeks after the highest incidences. In contrast, standard care hospitals transfer their patients at the time of highest SARS-CoV-2 case numbers. Two incidence peaks were accompanied by two peaks of increased secondary transport. Our findings show that interhospital transfers of SARS-CoV-2 and non-SARS-CoV-2 patients differ and that different hospital care levels initiated secondary transports at different times during the pandemic.

The analysis of the function of acylamino acid-releasing enzymes (AARE) in Physcomitrella and Arabidopsis reveals a connection between AARE and plant ageing, suggesting a unified concept of ageing may exist across domains of life.

Abstract Objective To review our experience with a standardized dental management approach in patients with planned radiotherapy of the head and neck region based on preradiation and follow‐up data. Material and Methods Records of patients who underwent radiotherapy between June 2016 and November 2020 were reviewed. Data on dental findings and therapeutic recommendations were extracted from a prospectively managed database. Hospital records were used to obtain follow‐up data. Results Two hundred eighty‐one patient records were identified. After the exclusion of 81 patients because of incomplete data, 200 patients were included in the study. Dental findings relevant to radiotherapy were found in 144 cases (72.0%). Teeth extractions were recommended in 112 (56.0%) patients. Follow‐up data were available for 172 (86.0%) patients (mean follow‐up: 16.8 ± 10.7 months). Radiodermatitis was the most frequently observed sequela of radiotherapy (42.4%), followed by dysphagia (38.4%) and stomatitis (36.6%). Osteoradionecrosis was observed in only 2.3% of the patients. Conclusion Dental findings relevant to planned radiotherapy were frequent and in many cases resulted in recommendations for teeth extraction. Based on our standardized dental management protocol, we observed low rates of late oral complications after radiotherapy of the head and neck region.

Abstract 18F-FDG-PET/CT is standard to assess response in Hodgkin lymphoma by quantifying metabolic activity with the Deauville score. PET/CT, however, is time-consuming, cost-extensive, linked to high radiation and has a low availability. As an alternative, we investigated radiomics from non-contrast-enhanced computed tomography (NECT) scans. 75 PET/CT examinations of 43 patients on two different scanners were included. Target lesions were classified as Deauville score 4 positive (DS4+) or negative (DS4−) based on their SUVpeak and then segmented in NECT images. From these segmentations, 107 features were extracted with PyRadiomics. All further statistical analyses were then performed scanner-wise: differences between DS4+ and DS4− manifestations were assessed with the Mann–Whitney-U-test and single feature performances with the ROC-analysis. To further verify the reliability of the results, the number of features was reduced using different techniques. The feature median showed a high sensitivity for DS4+ manifestations on both scanners (scanner A: 0.91, scanner B: 0.85). It furthermore was the only feature that remained in both datasets after applying different feature reduction techniques. The feature median from NECT concordantly has a high sensitivity for DS4+ Hodgkin manifestations on two different scanners and thus could provide a surrogate for increased metabolic activity in PET/CT.

Abstract Background In radiomics studies, differences in the volume of interest (VOI) are often inevitable and may confound the extracted features. We aimed to correct this confounding effect of VOI variability by applying parametric maps with a fixed voxel size. Methods Ten scans of a cup filled with sodium chloride solution were scanned using a multislice computed tomography (CT) unit. Sphere-shaped VOIs with different diameters (4, 8, or 16 mm) were drawn centrally into the phantom. A total of 93 features were extracted conventionally from the original images using PyRadiomics. Using a self-designed and pretested software tool, parametric maps for the same 93 features with a fixed voxel size of 4 mm3 were created. To retrieve the feature values from the maps, VOIs were copied from the original images to preserve the position. Differences in feature quantities between the VOI sizes were tested with the Mann-Whitney U-test and agreement with overall concordance correlation coefficients (OCCC). Results Fifty-five conventionally extracted features were significantly different between the VOI sizes, and none of the features showed excellent agreement in terms of OCCCs. When read from the parametric maps, only 8 features showed significant differences, and 3 features showed an excellent OCCC (≥ 0.85). The OCCCs for 89 features substantially increased using the parametric maps. Conclusions This phantom study shows that converting CT images into parametric maps resolves the confounding effect of VOI variability and increases feature reproducibility across VOI sizes.

Abstract Background The anti‐coronavirus disease 2019 (COVID‐19) vaccines are of paramount importance in the fight against the COVID‐19 pandemic. Both viral vector‐ and nucleic acid‐based vaccines are known to effectively induce protection against the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) virus by generating high antibody titers and effective T‐cell responses to the spike protein they encode. Although these vaccines are being applied worldwide and have been extensively investigated, the immunomorphological events at the vaccination site with respect to SARS‐CoV‐2 spike protein expression have not yet been described. Methods We had the opportunity to examine the deltoid muscles of three men who died shortly after vaccination for unrelated reasons. We examined the vaccination sites histologically and immunohistochemically with various antibodies. Furthermore we incubated two different cell lines with one vaccine and examined the expression of the spike protein. Results The vaccination sites show a dense lymphohistiocytic interstitial infiltrate which surrounds the small vessels and extends into the perimysium. The spike protein is expressed by histiocytic cells with a dendritic shape that are CD68‐positive and CD207‐negative, fibrocytes, and very rare S100‐positive cells. Interestingly, the skeletal muscle, being constitutively human leukocyte antigen (HLA)‐A,B,C‐negative, is induced at different levels in each specimen. In a cell culture experiment, we confirmed the ability of fibroblasts and interdigitating dendritic sarcoma cells to express spike protein in vitro after incubation with the Comirnaty vaccine. Conclusions Histiocytic cells and fibrocytes are the heralds of spike protein synthesis at the vaccination site. The underlying cause of this apparent cell specifity is unknown. This needs to be investigated in future experiments, for example in an animal model.

Abstract Background The transfer of classic concepts of competency-based medical education into clinical practice has been proven to be difficult in the past, being described as partially fragmented, misleading and inadequate. At the beginning of training, novice doctors commonly feel overwhelmed, overloaded and exposed to extreme time pressure. The discrepancy between expected and actual clinical competence of doctors at the start of their speciality training jeopardizes patient safety. The framework of Entrustable Professional Activities (EPAs) is a promising instrument to effectively integrate competency-based training into clinical practice and may help to close this gap and consequently to improve patient safety. Methods For anaesthesiology, we developed 5 EPAs for final-year medical students. The EPAs comprised the following seven categories: 1. Title, 2. Specifications, 3. Limitations, 4. Competency domains, 5. Knowledge, abilities and skills, professional attitudes, 6. Assessment and 7. Entrustment. Based on a modified, online-based Delphi study, we further developed and refined these EPAs. Education experts were recruited from the alumni network of the Master of Medical Education (MME) degree course from the University of Heidelberg, Germany. Results 28 data sets were evaluated in three Delphi rounds. 82% of study participants had previous experience with EPAs. Qualitative and quantitative data formed the basis during the iterative process and resulted in complete descriptions of 5 EPAs for final-year medical students in anaesthesiology. Conclusions Our study including the associated description of 5 EPAs represent a further step and starting point for EPA-based curricula in medical training in Germany linking undergraduate training, to residency training and continuous medical education.

Ruiz-Molina et al. design a synthetic protein, MFHR13, as a multi-target complement regulator. They demonstrate that it protects sheep erythrocytes from complement attack exhibiting 26 and 4-fold the regulatory activity of eculizumab and human Factor H, respectively, which highlights its therapeutic potential.

The solid foundation laid by Prof Theophilus Oladapo Ogunlesi and other founding fathers of the National Postgraduate Medical College of Nigeria (NPMCN) has consistently achieved its mandate of producing specialist medical practitioners in Nigeria. The ever-changing world requires that we have in Nigeria a critical mass of subspecialists to tackle the various old and emerging ailments that afflict the citizenry. Subspecialist training is popular among Nigerian specialists. While those who are not yet subspecialists crave for the training. Nigerian-born sub-specialists in the diaspora are willing to assist in the training. It is now left to policy makers and captains of the training institutions to seize the chance and move us forward with a view to improving the current embarrassingly low number of subspecialists in Nigeria. The NPMCN is hereby beckoned to take the lead. In doing so, it should combine excellence with relevance.

The spatial organization of cell surface receptors is critical for cell signaling and drug action. Here, the authors develop an optoproteomic method for mapping surface protein interactions, revealing cellular responses to antibodies, drugs and viral particles as well as immunosynapse signaling events.

The aim of this study was to develop a deep learning-based algorithm for fully automated spleen segmentation using CT images and to evaluate the performance in conditions directly or indirectly affecting the spleen (e.g., splenomegaly, ascites). For this, a 3D U-Net was trained on an in-house dataset (n = 61) including diseases with and without splenic involvement (in-house U-Net), and an open-source dataset from the Medical Segmentation Decathlon (open dataset, n = 61) without splenic abnormalities (open U-Net). Both datasets were split into a training (n = 32.52%), a validation (n = 9.15%) and a testing dataset (n = 20.33%). The segmentation performances of the two models were measured using four established metrics, including the Dice Similarity Coefficient (DSC). On the open test dataset, the in-house and open U-Net achieved a mean DSC of 0.906 and 0.897 respectively (p = 0.526). On the in-house test dataset, the in-house U-Net achieved a mean DSC of 0.941, whereas the open U-Net obtained a mean DSC of 0.648 (p < 0.001), showing very poor segmentation results in patients with abnormalities in or surrounding the spleen. Thus, for reliable, fully automated spleen segmentation in clinical routine, the training dataset of a deep learning-based algorithm should include conditions that directly or indirectly affect the spleen.

We aimed to evaluate the stability of radiomic features in the liver of healthy individuals across different three-dimensional regions of interest (3D ROI) sizes in T1-weighted (T1w) and T2-weighted (T2w) images from different MR scanners. We retrospectively included 66 examinations of patients without known diseases or pathological imaging findings acquired on three MRI scanners (3 Tesla I: 25 patients, 3 Tesla II: 19 patients, 1.5 Tesla: 22 patients). 3D ROIs of different diameters (10, 20, 30 mm) were drawn on T1w GRE and T2w TSE images into the liver parenchyma (segment V–VIII). We extracted 93 radiomic features from the different ROIs and tested features for significant differences with the Mann–Whitney-U (MWU)-test. The MWU-test revealed significant differences for most second- and higher-order features, indicating a systematic difference dependent on the ROI size. The features mean, median, root mean squared (RMS), 10th percentile, and 90th percentile were not significantly different. We also assessed feature robustness to ROI size variation with overall concordance correlation coefficients (OCCCs). OCCCs across the different ROI-sizes for mean, median, and RMS were excellent (>0.90) in both sequences on all three scanners. These features, therefore, seem robust to ROI-size variation and suitable for radiomic studies of liver MRI.

Aim was to develop a user-friendly method for creating parametric maps that would provide a comprehensible visualization and allow immediate quantification of radiomics features. For this, a self-explanatory graphical user interface was designed, and for the proof of concept, maps were created for CT and MR images and features were compared to those from conventional extractions. Especially first-order features were concordant between maps and conventional extractions, some even across all examples. Potential clinical applications were tested on CT and MR images for the differentiation of pulmonary lesions. In these sample applications, maps of Skewness enhanced the differentiation of non-malignant lesions and non-small lung carcinoma manifestations on CT images and maps of Variance enhanced the differentiation of pulmonary lymphoma manifestations and fungal infiltrates on MR images. This new and simple method for creating parametric maps makes radiomics features visually perceivable, allows direct feature quantification by placing a region of interest, can improve the assessment of radiological images and, furthermore, can increase the use of radiomics in clinical routine.

Orexins are neuropeptides originating from the hypothalamus that serve broad physiological roles, including the regulation of autonomic function, sleep-wake states, arousal and breathing. Lack of orexins may lead to narcolepsy and sleep disordered breathing. Orexinergic hypothalamic neurons send fibers to Kӧlliker-Fuse (KF) neurons that directly project to the rostroventral respiratory group, and phrenic and hypoglossal motor neurons. These connections indicate a potential role of orexin-modulated KF neurons in functionally linking the control of wakefulness/arousal and respiration. In a reduced preparation of juvenile rats Orexin B microinjected into the KF led to a transient increase in respiratory rate and hypoglossal output, however Orexin B modulation of the KF in intact preparations has not been explored. Here, we performed microinjections of the Orexin B mouse peptide and the synthetic Orexin 2 receptor agonist, MDK 5220, in the KF of spontaneously breathing, isoflurane anesthetized wild type mice. Microinjection of Orexin-2 receptor agonists into the KF led to transient slowing of respiratory rate, which was more exaggerated in response to Orexin-B than MDK 5220 injections. Our data suggest that Orexin B signaling in the KF may contribute to arousal-mediated respiratory responses.

Top et al. report the expression of a human blood-clotting factor IX-encoding cDNA in the moss Physcomitrium patens and demonstrate that in addition to the unspliced full-length transcript, the splicing machinery produced several different transcripts, and thus different protein isoforms, which the authors designate as heterosplicing. The authors present a web application, physCO, to automatize the process of P. patens-specific codon optimization, a tool that can assist in maximized functional protein amounts for biotechnology.

We aimed to evaluate radiomic features’ stability across different region of interest (ROI) sizes in CT and MR images. We chose a phantom with a homogenous internal structure so no differences for a feature extracted from ROIs of different sizes would be expected. For this, we scanned a plastic cup filled with sodium chloride solution ten times in CT and per MR sequence (T1-weighted-gradient-echo and T2-weighted-turbo-inversion-recovery-magnitude). We placed sphere-shaped ROIs of different diameters (4, 8, and 16 mm, and 4, 8, and 16 pixels) into the phantom’s center. Features were extracted using PyRadiomics. We assessed feature stability across ROI sizes with overall concordance correlation coefficients (OCCCs). Differences were tested for significance with the Mann–Whitney U-test. Of 93 features, 87 T1w-derived, 87 TIRM-derived, and 70 CT-derived features were significantly different between ROI sizes. Among MR-derived features, OCCCs showed excellent (>0.90) agreement for mean, median, and root mean squared for ROI sizes between 4 and 16 mm and pixels. We further observed excellent agreement for 10th and 90th percentile in T1w and 10th percentile in T2w TIRM images. There was no excellent agreement among the OCCCs of CT-derived features. In summary, many features indicated significant differences and only few showed excellent agreement across varying ROI sizes, although we examined a homogenous phantom. Since we considered a small phantom in an experimental setting, further studies to investigate this size effect would be necessary for a generalization. Nevertheless, we believe knowledge about this effect is crucial in interpreting radiomics studies, as features that supposedly discriminate disease entities may only indicate a systematic difference in ROI size.

Impaired chemoreflex responses are a central feature of opioid-induced respiratory depression, however, the mechanism through which mu opioid receptor agonists lead to diminished chemoreflexes is not fully understood. One brainstem structure involved in opioid-induced impairment of chemoreflexes is the nucleus of the solitary tract (NTS), which contains a population of neurons that express mu opioid receptors. Here, we tested whether caudal NTS neurons activated during the chemoreflex challenge express mu opioid receptors and overlap with neurons activated by opioids. Using genetic labeling of mu opioid receptor-expressing neurons and cFos immunohistochemistry as a proxy for neuronal activation, we examined the distribution of activated NTS neurons following hypercapnia, hypoxia, and morphine administration. The main finding was that hypoxia and hypercapnia primarily activated NTS neurons that did not express mu opioid receptors. Furthermore, concurrent administration of morphine with hypercapnia induced cFos expression in non-overlapping populations of neurons. Together these results suggest an indirect effect of opioids within the NTS, which could be mediated through mu opioid receptors on afferents and/or inhibitory interneurons.

Understanding SARS-CoV-2 breakthrough infections in vaccinated healthcare workers is of key importance in mitigating the effects of the COVID-19 pandemic in healthcare facilities. An observational prospective cohort study was conducted in vaccinated employees with acute SARS-CoV-2 infection between October 2021 and February 2022. Serological and molecular testing was performed to determine SARS-CoV-2 viral load, lineage, antibody levels, and neutralizing antibody titers. A total of 571 (9.7%) employees experienced SARS-CoV-2 breakthrough infections during the enrolment period, of which 81 were included. The majority (n = 79, 97.5%) were symptomatic and most (n = 75, 92.6%) showed Ct values < 30 in RT-PCR assays. Twenty-four (30%) remained PCR-positive for > 15 days. Neutralizing antibody titers were strongest for the wildtype, intermediate for Delta, and lowest for Omicron variants. Omicron infections occurred at higher anti-RBD-IgG serum levels (p = 0.00001) and showed a trend for higher viral loads (p = 0.14, median Ct difference 4.3, 95% CI [−2.5–10.5]). For both variants, viral loads were significantly higher in participants with lower anti-RBD-IgG serum levels (p = 0.02). In conclusion, while the clinical course of infection with both the Omicron and Delta variants was predominantly mild to moderate in our study population, waning immune response over time and prolonged viral shedding were observed.

The geochemical composition of glauconitic clasts is a provenance marker to distinguish the origin of gravity flows in sedimentary basins. Their geochemical variation is visible in major and trace elements composition, where the former can be visualized by element mapping of glauconitic clasts. By applying automated quantitative mineralogy on a Scanning Electron Microscope equipped with the ZEISS Mineralogic™ software platform, we developed a new way to visualize the element distribution in selected minerals, while masking out the other minerals simultaneously. This software applies energy dispersive spectroscopy spectrum deconvolution for each analysis point, therefore quantitative concentrations (wt%) of each element are determined for each pixel and visualized in the false-coloured element map with reproducible results for individual grains and zonations in these grains. The investigated glauconitic clasts were collected from eleven drill cores, covering a 1400 m-depth interval, and crosscutting four different gravity flow members. The clasts show three different trends: first, cores of glauconitic clasts show a variation of compositions that are mainly dependent on the conditions during their formation. This is most visible for Si and Al. Secondly, the composition of the glauconitic clasts changes with depth, their cores become first more, than less Fe-rich and more K-rich. This is probably an alteration effect, depending on temperature, and therefore mineral stability. And, thirdly, most glauconitic clasts are zoned, where rims are richer in Al and poorer in Fe and K. The visualization of the chemical variation in the glauconitic clasts proofs to be a useful tool to separate these conflicting element exchange processes.

Abstract Aims Heart failure (HF) is frequent in patients with acute ischaemic stroke (AIS) and associated with higher morbidity and mortality. Assessment of cardiac function in AIS patients using cardiovascular MRI (CMR) may help to detect HF. We report the rate of systolic and diastolic dysfunction in a cohort of patients with AIS using CMR and compare cine real‐time (CRT) sequences with the reference of segmented cine steady‐state free precession sequences. Methods and results Patients with AIS without known atrial fibrillation were prospectively enrolled in the HEart and BRain Interfaces in Acute Ischemic Stroke (HEBRAS) study (NCT 02142413) and underwent CMR at 3 Tesla within 7 days after AIS. Validity of CRT sequences was determined in 50 patients. A total of 229 patients were included in the analysis (mean age 66 years; 35% women; HF 2%). Evaluation of cardiac function was successful in 172 (75%) patients. Median time from stroke onset to CMR was 82 h (interquartile range 56–111) and 54 h (interquartile range 31–78) from cerebral MRI to CMR. Systolic dysfunction was observed in 43 (25%) and diastolic dysfunction in 102 (59%) patients. Diagnostic yield was similar using CRT or segmented cine imaging (no significant difference in left ventricular ejection fraction, myocardial mass, time to peak filling rate, and peak filling rate ratio E/A). Intraobserver and interobserver agreement was high (κ = 0.78–1.0 for all modalities). Conclusions Cardiovascular MRI at 3 Tesla is an appropriate method for the evaluation of cardiac function in a selected cohort of patients with AIS. Systolic and diastolic dysfunction is frequent in these patients. CRT imaging allows reliable assessment of systolic and diastolic function.

As biopharmaceuticals, recombinant proteins have become indispensable tools in medicine. An increasing demand, not only in quantity but also in diversity, drives the constant development and improvement of production platforms. The N-glycosylation pattern on biopharmaceuticals plays an important role in activity, serum half-life and immunogenicity. Therefore, production platforms with tailored protein N-glycosylation are of great interest. Plant-based systems have already demonstrated their potential to produce pharmaceutically relevant recombinant proteins, although their N-glycan patterns differ from those in humans. Plants have shown great plasticity towards the manipulation of their glycosylation machinery, and some have already been glyco-engineered in order to avoid the attachment of plant-typical, putatively immunogenic sugar residues. This resulted in complex-type N-glycans with a core structure identical to the human one. Compared to humans, plants lack the ability to elongate these N-glycans with β1,4-linked galactoses and terminal sialic acids. However, these modifications, which require the activity of several mammalian enzymes, have already been achieved for Nicotiana benthamiana and the moss Physcomitrella. Here, we present the first step towards sialylation of recombinant glycoproteins in Physcomitrella, human β1,4-linked terminal N-glycan galactosylation, which was achieved by the introduction of a chimeric β1,4-galactosyltransferase (FTGT). This chimeric enzyme consists of the moss α1,4-fucosyltransferase transmembrane domain, fused to the catalytic domain of the human β1,4-galactosyltransferase. Stable FTGT expression led to the desired β1,4-galactosylation. However, additional pentoses of unknown identity were also observed. The nature of these pentoses was subsequently determined by Western blot and enzymatic digestion followed by mass spectrometric analysis and resulted in their identification as α-linked arabinoses. Since a pentosylation of β1,4-galactosylated N-glycans was reported earlier, e.g., on recombinant human erythropoietin produced in glyco-engineered Nicotiana tabacum, this phenomenon is of a more general importance for plant-based production platforms. Arabinoses, which are absent in humans, may prevent the full humanization of plant-derived products. Therefore, the identification of these pentoses as arabinoses is important as it creates the basis for their abolishment to ensure the production of safe biopharmaceuticals in plant-based systems.

Fish embryos may be vulnerable to seawater acidification resulting from anthropogenic carbon dioxide (CO2) emissions or from excessive biological CO2 production in aquaculture systems. This study investigated CO2 effects on embryos of the European eel (Anguilla anguilla), a catadromous fish that is considered at risk from climate change and that is targeted for hatchery production to sustain aquaculture of the species. Eel embryos were reared in three independent recirculation systems with different pH/CO2 levels representing “control” (pH 8.1, 300 μatm CO2), end-of-century climate change (“intermediate”, pH 7.6, 900 μatm CO2) and “extreme” aquaculture conditions (pH 7.1, 3000 μatm CO2). Sensitivity analyses were conducted at 4, 24, and 48 hours post-fertilization (hpf) by focusing on development, survival, and expression of genes related to acute stress response (crhr1, crfr2), stress/repair response (hsp70, hsp90), water and solute transport (aqp1, aqp3), acid-base regulation (nkcc1a, ncc, car15), and inhibitory neurotransmission (GABAAα6b, Gabra1). Results revealed that embryos developing at intermediate pH showed similar survival rates to the control, but egg swelling was impaired, resulting in a reduction in egg size with decreasing pH. Embryos exposed to extreme pH had 0.6-fold decrease in survival at 24 hpf and a 0.3-fold change at 48 compared to the control. These observed effects of acidification were not reflected by changes in expression of any of the here studied genes. On the contrary, differential expression was observed along embryonic development independent of treatment, indicating that the underlying regulating systems are under development and that embryos are limited in their ability to regulate molecular responses to acidification. In conclusion, exposure to predicted end-of-century ocean pCO2 conditions may affect normal development of this species in nature during sensitive early life history stages with limited physiological response capacities, while extreme acidification will negatively influence embryonic survival and development under hatchery conditions.

As of late 2019, the COVID-19 pandemic has been a challenge to health care systems worldwide. Rapidly rising local COVID-19 incidence rates, result in demand for high hospital and intensive care bed capacities on short notice. A detailed up-to-date regional surveillance of the dynamics of the pandemic, precise prediction of required inpatient capacities of care as well as a centralized coordination of the distribution of regional patient fluxes is needed to ensure optimal patient care. In March 2020, the German federal state of Saxony established three COVID-19 coordination centers located at each of its maximum care hospitals, namely the University Hospitals Dresden and Leipzig and the hospital Chemnitz. Each center has coordinated inpatient care facilities for the three regions East, Northwest and Southwest Saxony with 36, 18 and 29 hospital sites, respectively. Fed by daily data flows from local public health authorities capturing the dynamics of the pandemic as well as daily reports on regional inpatient care capacities, we established the information and prognosis tool DISPENSE. It provides a regional overview of the current pandemic situation combined with daily prognoses for up to seven days as well as outlooks for up to 14 days of bed requirements. The prognosis precision varies from 21% and 38% to 12% and 15% relative errors in normal ward and ICU bed demand, respectively, depending on the considered time period. The deployment of DISPENSE has had a major positive impact to stay alert for the second wave of the COVID-19 pandemic and to allocate resources as needed. The application of a mathematical model to forecast required bed capacities enabled concerted actions for patient allocation and strategic planning. The ad-hoc implementation of these tools substantiates the need of a detailed data basis that enables appropriate responses, both on regional scales in terms of clinic resource planning and on larger scales concerning political reactions to pandemic situations.

As a part of a major reform of the medical curriculum in Germany, the national catalogue of learning objectives is being revised with the focus shifting from theory-based learning to teaching practical skills. Therefore, we conducted an online survey to answer the question, which practical skills are essential in anesthesia. Participants were asked to rate the relevance of several skills, that medical students should be able to perform at the time of graduation. A total of 2898 questionnaires could be evaluated. The highest ratings were made for “bringing a patient into lateral recumbent position” and “diagnosing a cardiac arrest”. All learning objectives regarding regional anesthesia were rated as irrelevant. Furthermore, learning objectives like “performing a bronchoscopy” or “performing a rapid sequence induction” had low ratings. In the subgroup analysis, physicians with advanced training and those who were working at university hospitals rated most skills with higher relevance compared to others. Our survey provides a good prioritization of practical skills for the development of new curricula and assessment frameworks. The results can also help to establish our discipline as a cross-sectional subject in competency-based medical education, thus further increasing the attractiveness for medical students.

Inflammatory bowel disease (IBD) is the most common form of intestinal inflammation associated with a dysregulated immune system response to the commensal microbiota in a genetically susceptible host. IBD includes ulcerative colitis (UC) and Crohn’s disease (CD), both of which are remarkably heterogeneous in their clinical presentation and response to treatment. This translates into a notable diagnostic challenge, especially in underdeveloped countries where IBD is on the rise and access to diagnosis or treatment is not always accessible for chronic diseases. The present work characterized, for the first time in our region, epigenetic biomarkers and gut microbial profiles associated with UC and CD patients in the Buenos Aires Metropolitan area and revealed differences between non-IBD controls and IBD patients. General metabolic functions associated with the gut microbiota, as well as core microorganisms within groups, were also analyzed. Additionally, the gut microbiota analysis was integrated with relevant clinical, biochemical and epigenetic markers considered in the follow-up of patients with IBD, with the aim of generating more powerful diagnostic tools to discriminate phenotypes. Overall, our study provides new insights into data analysis algorithms to promote comprehensive phenotyping tools using quantitative and qualitative analysis in a transkingdom interactions network context.

Abstract Background The aim of our study was the identification of genetic variants associated with postoperative complications after cardiac surgery. Methods We conducted a prospective, double-blind, multicenter, randomized trial (RIPHeart). We performed a genome-wide association study (GWAS) in 1170 patients of both genders (871 males, 299 females) from the RIPHeart-Study cohort. Patients undergoing non-emergent cardiac surgery were included. Primary endpoint comprises a binary composite complication rate covering atrial fibrillation, delirium, non-fatal myocardial infarction, acute renal failure and/or any new stroke until hospital discharge with a maximum of fourteen days after surgery. Results A total of 547,644 genotyped markers were available for analysis. Following quality control and adjustment for clinical covariate, one SNP reached genome-wide significance (PHLPP2, rs78064607, p = 3.77 × 10− 8) and 139 (adjusted for all other outcomes) SNPs showed promising association with p

The significance of mineralogical maturity as a provenance indicator has long been debated and we use this study to demonstrate that it can indeed be a powerful tool to track the distribution of sandstone reservoirs. We investigate the cause of the pronounced geographic and stratigraphic differences in mineralogical composition that are found in the Upper Triassic–Lower Jurassic Gassum Formation across the Norwegian–Danish Basin and surrounding areas. Zircon U-Pb dating of 46 sandstone samples including analysis of 4816 detrital grains are combined with quantifications of the detrital mineralogical composition and placed in a sequence stratigraphic framework. The results show that the Gassum Formation can be divided into a southeastern region with high mineralogical maturity and a less mature region to the northwest with more feldspars, rock fragments, micas, and heavy minerals. Both the mineralogical assemblage and the provenance signature have been thoroughly homogenized in the SE region where sediment supplies from the Fennoscandian Shield and the Variscan Orogen are evident. In the NW region, sediment was initially supplied from Fennoscandia only, but the provenance abruptly changed from the Telemarkia Terrane to comprising also the more distant Caledonian Orogen resulting in a different mineralogical assemblage. The change occurred during a basinwide regression and may be caused by tectonic movements in the hinterland that permanently changed the composition of the sediment supplied to the basin.

BackgroundPercutaneous hepatic melphalan perfusion (PHMP) for the selective treatment of hepatic metastases is known to be associated with procedural hypotension and coagulation disorders. Studies on anesthetic management, perioperative course, complications, and postoperative recovery in the intensive care unit (ICU) have not been published.MethodsIn a retrospective observational study, we analyzed consecutive patients who were admitted for PHMP over a 6-year period (2016-2021). Analyses included demographic, treatment, and outcome data with regard to short-term complications until ICU discharge.ResultsFifty-three PHMP procedures of 16 patients were analyzed. In all of the cases, procedure-related hypotension required the median (range) highest noradrenaline infusion rate of 0.5 (0.17-2.1) μg kg min-1 and fluid resuscitation volume of 5 (3-14) liters. Eighty-four PHMP-related complications were observed in 33 cases (62%), of which 9 cases (27%) involved grade III and IV complications. Complications included airway constriction (requiring difficult airway management), vascular catheterization issues (which resulted in the premature termination of PHMP, as well as to the postponement of PHMP and to the performance of endovascular bleeding control after PHMP), and renal failure that required hemodialysis. Discharge from the ICU was possible after one day in most cases (n = 45; 85%); however, in 12 cases (23%), prolonged mechanical ventilation was required. There were no procedure-related fatalities.ConclusionsPHMP is frequently associated with challenging cardiovascular conditions and complications that require profound anesthetic skills. For safety reasons, PHMP should only be performed in specialized centers that provide high-level hospital infrastructures and interdisciplinary expertise.

Background Extracorporeal Membrane Oxygenation (ECMO) use is increasing despite limited evidence. The aim of this study was to demonstrate heterogeneity of ECMO use and its association with hospital size and annual frequency in Germany. Methods This is a database analysis of all ECMO cases in Germany from 2010 to 2016 using the German Diagnosis Related Groups (DRG) coding system for ECMO. Results During the study period, 510 hospitals performed 29,929 ECMO runs (12,572 vvECMO, 11,504 vaECMO, 1993 pECLA) with an increase over time. Mortality ranged between 58% and 66% for vaECMO cases and 66% and 53% for vvECMO cases. 304 (61%) hospitals performed only one ECMO per year. 78%% of all ECMO runs were performed in centres with more than 20 cases per year and more than half of all ECMO runs were performed in hospitals with >1.000 beds. Mortality for vv and vaECMO was highest in very small hospitals (< 200 beds; 70%; 74%) and very large hospitals (>1000 beds; 60%; 62%). Conclusions Use of ECMO is still increasing and a substantial proportion of hospitals performs very few ECMO runs. Small hospitals had a significantly higher mortality, but dependence on hospital size and ECMO mortality was irregular.

Recombinantly produced proteins are indispensable tools for medical applications. Since the majority of them are glycoproteins, their N-glycosylation profiles are major determinants for their activity, structural properties and safety. For therapeutical applications, a glycosylation pattern adapted to product and treatment requirements is advantageous. Physcomitrium patens (Physcomitrella, moss) is able to perform highly homogeneous complex-type N-glycosylation. Additionally, it has been glyco-engineered to eliminate plant-specific sugar residues by knock-out of the β1,2-xylosyltransferase and α1,3-fucosyltransferase genes (Δxt/ft). Furthermore, Physcomitrella meets wide-ranging biopharmaceutical requirements such as GMP compliance, product safety, scalability and outstanding possibilities for precise genome engineering. However, all plants, in contrast to mammals, lack the capability to perform N-glycan sialylation. Since sialic acids are a common terminal modification on human N-glycans, the property to perform N-glycan sialylation is highly desired within the plant-based biopharmaceutical sector. In this study, we present the successful achievement of protein N-glycan sialylation in stably transformed Physcomitrella. The sialylation ability was achieved in a Δxt/ft moss line by stable expression of seven mammalian coding sequences combined with targeted organelle-specific localization of the encoded enzymes responsible for the generation of β1,4-galactosylated acceptor N-glycans as well as the synthesis, activation, transport and transfer of sialic acid. Production of free (Neu5Ac) and activated (CMP-Neu5Ac) sialic acid was proven. The glycosidic anchor for the attachment of terminal sialic acid was generated by the introduction of a chimeric human β1,4-galactosyltransferase gene under the simultaneous knock-out of the gene encoding the endogenous β1,3-galactosyltransferase. Functional complex-type N-glycan sialylation was confirmed via mass spectrometric analysis of a stably co-expressed recombinant human protein.

A collection of 30 Phytophthora cactorum and 12 P. pseudotsugae (subclade 1a) strains isolated from several recent surveys across California was phylogenetically compared to a worldwide collection of 112 conspecific strains using sequences from three barcoding loci. The surveys baited P. cactorum from soil and water across a wide variety of forested ecosystems with a geographic range of more than 1000 km. Two cosmopolitan lineages were identified within the widespread P. cactorum, one being mainly associated with strawberry production and the other more closely associated with apple orchards, oaks and ornamental trees. Two other well-sampled P. cactorum lineages, including one that dominated Californian restoration outplantings, were only found in the western United States, while a third was only found in Japan. Coastal California forest isolates of both Phytophthora species exhibited considerable diversity, suggesting both may be indigenous to the state. Many isolates with sequence accessions deposited as P. cactorum were determined to be P. hedraiandra and P. ×serendipita, with one hybrid lineage appearing relatively common across Europe and Asia. This study contains the first report of P. pseudotsugae from the state of California and one of the only reports of that species since its original description.

INTRODUCTION:Posttraumatic pneumothorax (PTX) is often overseen in anteroposterior chest X-ray. Chest sonography and Electrical Impedance Tomography (EIT) can both be used at the bedside and may provide complementary information. We evaluated the performance of EIT for diagnosing posttraumatic PTX in a pig model. METHODS:This study used images from an existing database of images acquired from 17 mechanically ventilated pigs, which had sustained standardized blunt chest trauma and had undergone repeated thoracic CT and EIT. 100 corresponding EIT/CT datasets were randomly chosen from the database and anonymized. Two independent and blinded observers analyzed the EIT data for presence and location of PTX. Analysis of the corresponding CTs by a radiologist served as reference. RESULTS:87/100 cases had at least one PTX detected by CT. Fourty-two cases showed a PTX > 20% of the sternovertebral diameter (PTXtrans20), whereas 52/100 PTX showed a PTX>3 cm in the craniocaudal diameter (PTXcc3), with 20 cases showing both a PTXtranscc and a PTXcc3. We found a very low agreement between both EIT observers considering the classification overall PTX/noPTX (κ = 0.09, p = 0.183). For PTXtrans20, sensitivity was 59% for observer 1 and 17% for observer 2, with a specificity of 48% and 50%, respectively. For PTXcc3, observer 1 showed a sensitivity of 60% with a specificity of 51% while the sensitivity of observer 2 was 17%, with a specificity of 89%. By programming a semi-automatized detection algorithm, we significantly improved the detection rate of PTXcc3, with a sensitivity of 73% and a specificity of 70%. However, detection of PTXtranscc was not improved. CONCLUSION:In our analysis, visual interpretation of EIT without specific image processing or comparison with baseline data did not allow clinically useful diagnosis of posttraumatic PTX. Multimodal imaging approaches, technical improvements and image postprocessing algorithms might improve the performance of EIT for diagnosing PTX in the future.

Summary: CRISPR-Cas systems adapt their immunological memory against their invaders by integrating short DNA fragments into clustered regularly interspaced short palindromic repeat (CRISPR) loci. While Cas1 and Cas2 make up the core machinery of the CRISPR integration process, various class I and II CRISPR-Cas systems encode Cas4 proteins for which the role is unknown. Here, we introduced the CRISPR adaptation genes cas1, cas2, and cas4 from the type I-D CRISPR-Cas system of Synechocystis sp. 6803 into Escherichia coli and observed that cas4 is strictly required for the selection of targets with protospacer adjacent motifs (PAMs) conferring I-D CRISPR interference in the native host Synechocystis. We propose a model in which Cas4 assists the CRISPR adaptation complex Cas1-2 by providing DNA substrates tailored for the correct PAM. Introducing functional spacers that target DNA sequences with the correct PAM is key to successful CRISPR interference, providing a better chance of surviving infection by mobile genetic elements. : Kieper et al. demonstrate that the ubiquitous protein Cas4 assists Cas1 and Cas2 in the selection of new CRISPR spacers with a PAM licensing efficient CRISPR interference. Keywords: CRISPR adaptation, Cas4, spacer acquisition, type I-D CRISPR-Cas system

This study retrospectively analyzed the performance of artificial neural networks (ANN) to predict overall survival (OS) or locoregional failure (LRF) in HNSCC patients undergoing radiotherapy, based on 2-[18F]FDG PET/CT and clinical covariates. We compared predictions relying on three different sets of features, extracted from 230 patients. Specifically, (i) an automated feature selection method independent of expert rating was compared with (ii) clinical variables with proven influence on OS or LRF and (iii) clinical data plus expert-selected SUV metrics. The three sets were given as input to an artificial neural network for outcome prediction, evaluated by Harrell’s concordance index (HCI) and by testing stratification capability. For OS and LRF, the best performance was achieved with expert-based PET-features (0.71 HCI) and clinical variables (0.70 HCI), respectively. For OS stratification, all three feature sets were significant, whereas for LRF only expert-based PET-features successfully classified low vs. high-risk patients. Based on 2-[18F]FDG PET/CT features, stratification into risk groups using ANN for OS and LRF is possible. Differences in the results for different feature sets confirm the relevance of feature selection, and the key importance of expert knowledge vs. automated selection.

The mid- to late Holocene aridification pattern of NW Africa remains a matter of controversial debate. While many marine climate reconstructions indicate a relatively abrupt aridification at ∼5.5 ka BP, terrestrial palaeoclimate records rather show a spatially and temporally heterogeneous transition towards the modern arid state. To bridge conflicting evidence, we analysed high-resolution (sub-seasonal) ontogenetic oxygen isotope (δ18O) records and bulk 87Sr/86Sr ratios of bivalve shells and fish otoliths of estuarine-associated species. Samples were excavated from archaeological deposits formed during the mid- to late Holocene in a large palaeo-estuary east of the Banc d’Arguin, Mauritania. The mid-Holocene (∼5.2 ka BP) δ18O records indicated unrealistically high water temperatures when assuming a modern value for δ18OSeawater, suggesting a substantial input of isotopically lighter water to the study area. Respective salinity estimates consistently indicated persistent monsoonal discharge. Moreover, 87Sr/86Sr ratios of bivalve shells deviated considerably from the rather stable global seawater composition, further supporting the presence of significant terrestrial runoff between 5.0 and 5.3 ka BP. Altogether, our results support doubts regarding an abrupt termination of the African Humid Period (AHP) in the coastal areas of NW Africa, and show that fully marine conditions were established along the Banc d’Arguin by ∼3.0 ka BP.

Chronic kidney disease (CKD) is an increasing health burden (affecting approximately 13.4% of the population). Currently, no curative treatment options are available and treatment is focused on limiting the disease progression. The accumulation of senescent cells has been implicated in the development of kidney fibrosis by limiting tissue rejuvenation and through the secretion of pro-fibrotic and pro-inflammatory mediators termed as the senescence-associated secretory phenotype. The clearance of senescent cells in aging models results in improved kidney function, which shows promise for the options of targeting senescent cells in CKD. There are several approaches for the development of “senotherapies”, the most rigorous of which is the elimination of senescent cells by the so-called senolytic drugs either newly developed or repurposed for off-target effects in terms of selectively inducing apoptosis in senescent cells. Several chemotherapeutics and checkpoint inhibitors currently used in daily oncological practice show senolytic properties. However, the applicability of such senolytic compounds for the treatment of renal diseases has hardly been investigated. A serious concern is that systemic side effects will limit the use of senolytics for kidney fibrosis. Specifically targeting senescent cells and/or targeted drug delivery to the kidney might circumvent these side effects. In this review, we discuss the connection between CKD and senescence, the pharmacological options for targeting senescent cells, and the means to specifically target the kidney.

The human complement system is an important part of the immune system responsible for lysis and elimination of invading microorganisms and apoptotic body cells. Improper activation of the system due to deficiency, mutations, or autoantibodies of complement regulators, mainly factor H (FH) and FH-related proteins (FHRs), causes severe kidney and eye diseases. However, there is no recombinant FH therapeutic available on the market. The first successful recombinant production of FH was accomplished with the moss bioreactor, Physcomitrella patens. Recently, a synthetic regulator, MFHR1, was designed to generate a multitarget complement inhibitor that combines the activities of FH and the FH-related protein 1 (FHR1). The potential of MFHR1 was demonstrated in a proof-of-concept study with transiently transfected insect cells. Here, we present the stable production of recombinant glyco-engineered MFHR1 in the moss bioreactor. The key features of this system are precise genome engineering via homologous recombination, Good Manufacturing Practice-compliant production in photobioreactors, high batch-to-batch reproducibility, and product stability. Several potential biopharmaceuticals are being produced in this system. In some cases, these are even biobetters, i.e., the recombinant proteins produced in moss have a superior quality compared to their counterparts from mammalian systems as for example moss-made aGal, which successfully passed phase I clinical trials. Via mass spectrometry-based analysis of moss-produced MFHR1, we now prove the correct synthesis and modification of this glycoprotein with predominantly complex-type N-glycan attachment. Moss-produced MFHR1 exhibits cofactor and decay acceleration activities comparable to FH, and its mechanism of action on multiple levels within the alternative pathway of complement activation led to a strong inhibitory activity on the whole alternative pathway, which was higher than with the physiological regulator FH.

Direct catalytic methanation of CO2 (from CO2/CH4 biogas mixture) to produce biomethane was conducted in a pilot demonstration plant. In the demonstration project (MeGa-StoRE), a biogas desulfurization process and thermochemical methanation of biogas using hydrogen produced by water electrolysis were carried out at a fully operational biogas plant in Denmark. The main objective of this part of the project was to design and develop a reactor system for catalytic conversion of CO2 in biogas to methane and feed biomethane directly to the existing natural gas grid. A process was developed in a portable container with a 10 Nm3/h of biogas conversion capacity. A test campaign was run at a biogas plant for more than 6 months, and long-time operation revealed a stable steady-state conversion of more than 90% CO2 conversion to methane. A detailed catalytic study was performed to investigate the high activity and stability of the applied catalyst.

We report the modular synthesis of three different types of neutral κ2-P,N-ligands comprising an imine and a phosphine binding site. These ligands were reacted with rhodium, iridium and palladium metal precursors and the structures of the resulting complexes were elucidated by means of X-ray crystallography. We observed that subtle changes of the ligand backbone have a significant influence on the binding geometry und coordination properties of these bidentate P,N-donors.

Digestive system functionality of fish larvae relies on the onset of genetically pre-programmed and extrinsically influenced digestive functions. This study explored how algal supplementation (green-water) until 14 days post hatch (dph) and the ingestion of food [enriched rotifer (Brachionus plicatilis) paste] from 15 dph onward affects molecular maturation and functionality of European eel larval ingestion and digestion mechanisms. For this, we linked larval biometrics to expression of genes relating to appetite [ghrelin (ghrl), cholecystokinin (cck)], food intake [proopiomelanocortin (pomc)], digestion [trypsin (try), triglyceride lipase (tgl), amylase (amyl)], energy metabolism [ATP synthase F0 subunit 6 (atp6), cytochrome-c-oxidase 1 (cox1)], growth [insulin-like growth factor (igf1)] and thyroid metabolism [thyroid hormone receptors (thrαA, thrβB)]. Additionally, we estimated larval nutritional status via nucleic acid analysis during transition from endogenous and throughout the exogenous feeding stage. Results showed increased expression of ghrl and cck on 12 dph, marking the beginning of the first-feeding window, but no benefit of larviculture in green-water was observed. Moreover, expression of genes relating to protein (try) and lipid (tgl) hydrolysis revealed essential digestive processes occurring from 14 to 20 dph. On 16 dph, a molecular response to initiation of exogenous feeding was observed in the expression patterns of pomc, atp6, cox1, igf1, thrαA and thrβB. Additionally, we detected increased DNA contents, which coincided with increased RNA contents and greater body area, reflecting growth in feeding compared to non-feeding larvae. Thus, the here applied nutritional regime facilitated a short-term benefit, where feeding larvae were able to sustain growth and better condition than their non-feeding conspecifics. However, RNA:DNA ratios decreased from 12 dph onward, indicating a generally low larval nutritional condition, probably leading to the point-of-no-return and subsequent irreversible mortality due to unsuccessful utilization of exogenous feeding. In conclusion, this study molecularly identified the first-feeding window in European eel and revealed that exogenous feeding success occurs concurrently with the onset of a broad array of enzymes and hormones, which are known to regulate molecular processes in feeding physiology. This knowledge constitutes essential information to develop efficient larval feeding strategies and will hopefully provide a promising step toward sustainable aquaculture of European eel.