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Spencer E Thomas,1,2,* Jeffrey Comer,1,* Min Jung Kim,1 Shanna Marroquin,1 Vaibhav Murthy,1 Meghana Ramani,1 Tabetha Gaile Hopke,2 Jayden McCall,1 Seong-O Choi,1 Robert K DeLong1 1Nanotechnology Innovation Center of Kansas State, Department of Anatomy and Physiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA; 2Department of Biomedical Science, Missouri State University, Springfield, MO, USA *These authors contributed equally to this work Introduction: Biomedical applications of nanoparticles (NPs) as enzyme inhibitors have recently come to light. Oxides of metals native to the physiological environment (eg, Fe, Zn, Mg, etc.) are of particular interest—especially the functional consequences of their enzyme interaction. Materials and methods: Here, Fe2O3, zinc oxide (ZnO), magnesium oxide (MgO) and nickel oxide (NiO) NPs are compared to copper (Cu) and boron carbide (B4C) NPs. The functional impact of NP interaction to the model enzyme luciferase is determined by 2-dimensional fluorescence difference spectroscopy (2-D FDS) and 2-dimensional photoluminescence difference spectroscopy (2-D PLDS). By 2-D FDS analysis, the change in maximal intensity and in 2-D FDS area under the curve (AUC) is in the order Cu~B4C>ZnO>NiO>>Fe2O3>MgO. The induced changes in protein conformation are confirmed by tryptic digests and gel electrophoresis. Results: Analysis of possible trypsin cleavage sites suggest that cleavage mostly occurs in the range of residues 112–155 and 372–439, giving a major 45 kDa band. By 2-D PLDS, it is found that B4C NPs completely ablate bioluminescence, while Cu and Fe2O3 NPs yield a unique bimodal negative decay rate, -7.67×103 and -3.50×101 relative light units respectively. Cu NPs, in particular, give a remarkable 271% change in enzyme activity. Molecular dynamics simulations in water predicted that the surfaces of metal oxide NPs become capped with metal hydroxide groups under physiological conditions, while the surface of B4C becomes populated with boronic acid or borinic acid groups. These predictions are supported by the experimentally determined zeta potential. Thin layer chromatography patterns further support this conception of the NP surfaces, where stabilizing interactions were in the order ionic>polar>non-polar for the series tested. Conclusion: Overall the results suggest that B4C and Cu NP functional dynamics on enzyme biochemistry are unique and should be examined further for potential ramifications on other model, physiological or disease-relevant enzymes. Keywords: 2-dimensional fluorescence difference, 2-D FDS, AUC, corrected light intensity emitted, emission wavelength, excitation wavelength, wavelength of max intensity, RLU

Susan R Childs,1,* Emma M Casely,2,* Bianca M Kuehler,1 Stephen Ward,1 Charlotte L Halmshaw,1 Sarah E Thomas,1 Ian D Goodall,1 Carsten Bantel1,3 1Pain Medicine, Chelsea and Westminster Hospital NHS Foundation Trust, London, 2Anaesthetic Department, Hillingdon Hospital, Uxbridge, 3Section of Anaesthetics, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Chelsea and Westminster Hospital Campus, London, UK *These authors contributed equally to this manuscript Abstract: Recent research has confirmed that between 25% and 33% of all hospitalized patients experience unacceptable levels of pain. Studies further indicate that this reduces patient satisfaction levels, lengthens hospital stays, and increases cost. Hospitals are aiming to discharge patients earlier, and this can interfere with adequate pain management. Therefore, the pain service at Chelsea and Westminster Hospital has adapted to this changing model of care. An increasing body of evidence demonstrates that psychological factors are key components of patients’ pain experiences in both acute and chronic pain. Therefore, it is reasonable to suggest a clinical psychologist should be involved in inpatient pain management. This small study discusses three cases that highlight how patient care could be improved by including a clinical psychologist as part of the inpatient pain team. Two cases particularly highlight the active role of the psychologist in the diagnosis and management of common conditions such as fear and anxiety, along with other psychiatric comorbidities. The management therefore employed an eclectic approach adapted from chronic pain and comprising of behavioral, cognitive behavioral, and dialectical behavioral therapeutic techniques blended with brief counseling. The third case exemplifies the importance of nurse-patient interactions and the quality of nurse-patient relationships on patient outcomes. Here, the psychologist helped to optimize communication and to resolve a difficult and potentially risk-laden situation. This small case series discusses the benefits derived from the involvement of a clinical psychologist in the management of inpatient pain, and therefore illustrates the need for novel initiatives for inpatient pain services. However, future research is warranted to validate this approach. Keywords: acute pain, aggression, anxiety, borderline personality disorder, psychology

Abstract Background Task-shifting from primary care physicians (PCPs) to nurses is a means of overcoming PCP shortages and meeting the needs of patients receiving primary care. The aim of this overview of systematic reviews is to assess the effects of delegation or substitution of PCPs’ activities by nurses on patient relevant, clinical, professional and health services-related outcomes. Methods We conducted a systematic literature search for secondary literature in Medline, Embase, Pubmed, the Cochrane Library, and the Cumulative Index of Nursing and Allied Health Literature (CINAHL). We included systematic reviews and meta-analyses that analysed randomised controlled trials (RCTs) and controlled, prospective trials in English and German. Abstracts and full-text articles were screened independently by two reviewers. Full-text articles were assessed using the Overview Quality Assessment Questionnaire. After data extraction a narrative synthesis was performed. We defined patient-relevant outcomes as our primary outcomes. Results We included six systematic reviews. The interventions included first contact, history taking and assessment, patient education, review of drug treatment, referrals to GPs and other health professionals, ordering further investigations and ongoing care. Two meta-analyses showed a relative risk reduction of mortality in favour of nurse-led care, whereby the reduction in one analysis was significant. The effect was highest in the group of more highly qualified nurse practitioners (RR 0.19), as opposed to nurse practitioners (RR 0.76) and registered nurses (RR 0.92). Two meta-analyses showed a relative risk reduction in hospital admissions and patient satisfaction. Whereas care conducted by physicians and registered nurses led to the same outcomes, care conducted by nurse practitioners led to better outcomes (RR 0.74). An analysis according to nursing group showed that patients were more satisfied with treatment by registered nurses (SMD 1.37) than with treatment conducted by nurse practitioners and more qualified nurse practitioners (SMD 0.17). In terms of patient-relevant outcomes, no differences were observed between physician-led care and nurse-led care in terms of physical function, quality of life and pain. Conclusion Nurse-led care is probably as safe or safer than physician-led care in terms of mortality and hospital admissions. However, the impact of nursing staff training has not been sufficiently examined.

Abstract Background Ductal pancreatic adenocarcinoma (PDAC) still has a dismal prognosis even when deemed resectable. A cancer free resection margin (R0) is associated with a more favourable prognosis than the presence of tumour cells at resection margin (R1). However, the precise definition of the R0 status is still a matter of debate in PDAC. For a more accurate determination of R0 in PDAC the concept of circumferential resection margins (CRM) has been established and has been incorporated into the German national S3 guideline on exocrine pancreatic cancer. However, an international standardized nomenclature of CRM is still missing, and the clinical value of the CRM concept is not yet fully established. Here we evaluate whether the CRM status as defined in the national German S3 guideline corresponds with overall and progression free survival in PDAC using data from the regional cancer registry of the State of Baden Württemberg in Germany. Methods Data from the cancer registry of the State of Baden-Württemberg, Germany, were used to assess the relationship between CRM-status and progression free survival (PFS) as well as 3-year overall survival (OS) using documented patients diagnosed with resectable ductal adenocarcinoma of the pancreas between 2015 and 2020. Patients were residents of the State of Baden-Württemberg and underwent surgery for PDAC. The R-status was assessed according to the national German S3 guideline with R0 wide/CRM- when CRM is > 1 mm from the tumour, R0 narrow/CRM + when CRM is ≤ 1 mm from the tumour and R1 when tumour cells are found at the resection margin. Results In total we identified 1098 cases surgically treated for pancreatic cancer and fulfilling the inclusion criteria. 340 patients had an R0 wide/CRM- resection, 410 patients an R0 narrow/CRM + resection, and 348 patients an R1 resection. The R0 wide/CRM- status was associated with a significantly increased median OS rate compared to the other two groups (51,5%, 37,4% and 26,7% for R0 wide/CRM-, R0 narrow/CRM + and R1, respectively). mPFS was also longer in the R0 wide/CRM- group. These findings were robust with regards to grading and tumour location. Conclusions CRM is prognostic for patients with resectable PDAC making the pathological assessment of the R-status according to the CRM concept worthwhile.

Abstract Small open reading frames (smORFs) shorter than 100 codons are widespread and perform essential roles in microorganisms, where they encode proteins active in several cell functions, including signal pathways, stress response, and antibacterial activities. However, the ecology, distribution and role of small proteins in the global microbiome remain unknown. Here, we construct a global microbial smORFs catalog (GMSC) derived from 63,410 publicly available metagenomes across 75 distinct habitats and 87,920 high-quality isolate genomes. GMSC contains 965 million non-redundant smORFs with comprehensive annotations. We find that archaea harbor more smORFs proportionally than bacteria. We moreover provide a tool called GMSC-mapper to identify and annotate small proteins from microbial (meta)genomes. Overall, this publicly-available resource demonstrates the immense and underexplored diversity of small proteins.

Abstract Aims Right ventricular failure after left ventricular assist device (LVAD) implantation is a major concern that remains challenging to predict. We sought to investigate the relationship between preoperative pulmonary artery pulsatility index (PAPi) and mortality after LVAD implantation. Methods and results A retrospective analysis of the ASSIST‐ICD multicentre registry allowed the assessment of PAPi before LVAD according to the formula [(systolic pulmonary artery pressure − diastolic pulmonary artery pressure)/central venous pressure]. The primary endpoint was survival at 3 months, according to the threshold value of PAPi determined by the receiver operating characteristic (ROC) curve. A multivariate analysis including demographic, echographic, haemodynamic, and biological variables was performed to identify predictive factors for 2 year mortality. One hundred seventeen patients were included from 2007 to 2021. The mean age was 58.45 years (±13.16), with 15.4% of women (sex ratio 5.5). A total of 53.4% were implanted as bridge to transplant and 43.1% as destination therapy. Post‐operative right ventricular failure was observed in 57 patients (48.7%), with no significant difference between survivors and non‐survivors at 1 month (odds ratio 1.59, P = 0.30). The median PAPi for the whole study population was 2.83 [interquartile range 1.63–4.69]. The threshold value of PAPi determined by the ROC curve was 2.84. Patients with PAPi ≥ 2.84 had a higher survival rate at 3 months [PAPi

Abstract Background Social isolation and loneliness are urgent public health concerns associated with negative physical and mental health outcomes. Understanding effective remedies is crucial in addressing these problems. This umbrella review aimed to synthesize and critically appraise scientific evidence on the effectiveness of social isolation and loneliness interventions overall and across subgroups. We focused on systematic reviews (SRs) of randomized controlled trials (RCTs). Methods We searched seven databases (June 2022 and updated June 2023) and supplemented the search with grey literature and reference screening to identify SRs published since 2017. Screening, data extraction, and quality assessment using the AMSTAR2 tool were conducted independently by author pairs, with disagreements resolved through discussion. Results We included 29 SRs, 16 with meta-analysis and 13 with narrative synthesis. All SRs focused on loneliness, with 12 additionally examining social isolation. Four SRs focused on young people, 11 on all ages, and 14 on older adults. The most frequently examined intervention types were social (social contact, social support), psychological (therapy, psychoeducation, social skills training), and digital (e.g., computer use and online support). Meta-analyses indicated small-to-moderate beneficial effects, while narrative synthesis demonstrated mixed or no effect. Social interventions for social isolation and psychological interventions for loneliness were the most promising. However, caution is warranted due to the effects’ small magnitude, significant heterogeneity, and the variable quality of SRs. Digital and other interventions showed mixed or no effect; however, caution is advised in interpreting these results due to the highly diverse nature of the interventions studied. Conclusions This overview of SRs shows small to moderate effectiveness of social interventions in reducing social isolation and psychological ones in tackling loneliness. Further rigorously conducted RCTs and SRs are needed to guide policy decisions regarding the implementation of efficacious and scalable interventions. Evaluation should focus on both preventive structural interventions and tailored mitigating strategies that address specific types and causes of loneliness.

Exploring the capabilities of remote sensing technologies for identifying stress responses in trees due to environmental pressures is crucial for comprehension, management, and maintenance of forests that are productive, healthy, and resilient. In recent decades, research on forest health monitoring has been predominantly focused on data obtained remotely, either from satellites or crewed aircraft. During the last few years, Uncrewed Aerial Vehicles (UAVs) have gained prominence as invaluable remote sensing platforms, increasingly being employed for forest surveying. As intermediary between traditional remote sensing methods and ground-level observations, UAVs can capture high-resolution imagery from low altitudes, even below cloud cover, in unprecedented detail. This ability allows for the precise detection of stress responses at the individual tree scale. In our study, we have acquired a highly heterogenous, multispectral time-series dataset from the International Co-operative Programme on Assessment and Monitoring of Air Pollution Effects on Forests (ICP Forests) inventory plots across Bavaria, Germany, focusing on the main tree species. The data was recorded over three consecutive years from a UAV with the objective of monitoring tree physiological stress responses. Concurrently, with the drone flights, the ground-based forest condition surveying (Level-1 monitoring) was conducted, serving as ground-truth validation, and providing detailed information on tree health indicators, such as defoliation and discoloration. Our findings revealed that multispectral imagery obtained from a UAV closely aligns with field data, proving effective detection of physiological stress in trees. Remarkably, in conjunction to the red, red edge, and near-infrared band, the inclusion of the blue band emerged as a critical indicator of tree stress when incorporated into the Blue Normalized Difference Vegetation Index (BNDVI), depending on factors such as tree species, class division, and atmospheric conditions. Furthermore, the averaged values per sample tree over three years, alongside the 5th and 25th percentile of the data distribution, have proven to be of key importance. Based on spectral indices, we achieved good classification accuracies by training species-specific gradient boosting models (macro F1-scores ranging from 0.492 to 0.769). These models can assist in quantifying tree stress responses, thereby supporting the objectives of the ICP Forests program, potentially leading to substantial cost savings or increased coverage in the future.

Therapeutic antibodies (Ab) have revolutionized the management of multiple illnesses including respiratory tract infections (RTIs). However, anti-infectious Ab displayed several limitations including antigen restrictiveness, narrowed therapeutic windows, and limited dose in the vicinity of the target when delivered by parenteral routes. Strategies enhancing further Ab-dependent containment of infection are currently needed. Here we showed that a combination of inhaled anti-infectious Ab and probiotics is an efficient formulation to protect against lung infection. Using a mouse model of Pseudomonas aeruginosa-induced pneumonia, we demonstrated a synergistic effect reducing both bacterial burden and pro-inflammatory response affording protection against primary and secondary infections. This is the first study showing that the local combination in the airways of anti-infective Ab and probiotics subverts suboptimal potency of Ab monotherapy and provides protection against respiratory pathogen.

Abstract Poor treatment responses of pancreatic ductal adenocarcinoma (PDAC) are in large part due to tumor heterogeneity and an immunosuppressive desmoplastic tumor stroma that impacts interactions with cells in the tumor microenvironment (TME). Thus, there is a pressing need for models to probe the contributions of cellular and noncellular crosstalk. Organoids are promising model systems with the potential to generate a plethora of data including phenotypic, transcriptomic and genomic characterization but still require improvements in culture conditions mimicking the TME. Here, we describe an INTERaction with Organoid-in-MatriX ("InterOMaX") model system, that presents a 3D co-culture-based platform for investigating matrix-dependent cellular crosstalk. We describe its potential to uncover new molecular mechanisms of T cell responses to murine KPC (LSL-Kras G12D/+27 /Trp53 tm1Tyj/J /p48 Cre/+) PDAC cells as well as PDAC patient-derived organoids (PDOs). For this, a customizable matrix and homogenously sized organoid-in-matrix positioning of cancer cells were designed based on a standardized agarose microwell chip array system and established for co-culture with T cells and inclusion of stromal cells. We describe the detection and orthogonal analysis of murine and human PDAC cell populations with distinct sensitivity to T cell killing that is corroborated in vivo. By enabling both identification and validation of gene candidates for T cell resistance, this platform sets the stage for better mechanistic understanding of cancer cell-intrinsic resistance phenotypes in PDAC.

Objective Lipomatous hypertrophy of the interatrial septum (LHIS) is a distinct section of epicardial adipose tissue. However, its association with COPD is poorly documented. Methods Patients undergoing coronary computed tomography angiography (CTA) for clinical indications were recruited retrospectively and screened for LHIS and COPD. LHIS density and the coronary artery disease profile were quantified by CTA: stenosis severity (coronary artery disease radiological reporting system (CADRADS)), coronary artery calcium (CAC) and high-risk plaque (HRP). COPD patients with LHIS were matched for age and sex, the major cardiovascular risk factors (CVRFs), and compared to controls. Results The prevalence of LHIS in all 5466 patients was 5.9%. 151 (72.6%) of 208 patients with COPD had LHIS. LHIS density in COPD patients was higher (−10.93 HU versus −21.1 HU; p30 kg·m−2) patients (20.4 versus 13.6 HU; p=0.02). BMI was inversely correlated with LHIS density (BetaR −0.031; 95% CI: −0.054– −0.008; p=0.007). LHIS density was associated with COPD, but not with BMI on multivariate models. CAC and coronary stenosis severity (CADRADS and >50% stenosis) were not different (p=0.106, p=0.156 and p=0.350, respectively). HRPs were observed more frequently in COPD patients with severe Global Initiative for Chronic Obstructive Lung Disease (GOLD) stages ≥2 (32.3% versus 20.1%; p=0.044), but not when adding mild GOLD stages. Conclusions The prevalence of LHIS in COPD patients is high (72.6%), and the adipose tissue density is higher, indicating a higher brown fat component. In obese, patients LHIS density is lower and declines along with BMI. Coronary stenosis severity and calcium were not different; however HRPs were more frequent in severe COPD.

Summary: Pancreatic ductal adenocarcinoma (PDAC) presents significant challenges for targeted clinical interventions due to prevalent KRAS mutations, rendering PDAC resistant to RAF and MEK inhibitors (RAFi and MEKi). In addition, responses to targeted therapies vary between patients. Here, we explored the differential sensitivities of PDAC cell lines to RAFi and MEKi and developed an isogenic pair comprising the most sensitive and resistant PDAC cells. To simulate patient- or tumor-specific variations, we constructed cell-line-specific mechanistic models based on protein expression profiling and differential properties of KRAS mutants. These models predicted synergy between two RAFi with different conformation specificity (type I½ and type II RAFi) in inhibiting phospho-ERK (ppERK) and reducing PDAC cell viability. This synergy was experimentally validated across all four studied PDAC cell lines. Our findings underscore the need for combination approaches to inhibit the ERK pathway in PDAC.

In this study, fluid flow predictions using three different methods were compared: DeepCFD, an artificial intelligence code; computational fluid dynamics (CFD) using Ansys Fluent and OpenFOAM; and two-dimensional, two-component particle image velocimetry (PIV) measurements. The airfoils under investigation were the NACA 0012 with a 10° angle of attack and the NACA 6412 with a 0° angle of attack. To train DeepCFD, 763, 2585, and 6283 OpenFOAM simulations based on primitives were utilized. The investigation was conducted at a free stream velocity of 10 m/s and a Reynolds number of 82000. Results show that once the DeepCFD network is trained, prediction times are negligible, enabling real-time optimization of airfoils. The mean absolute error between CFD and DeepCFD, with 6283 trained primitives, for NACA 0012 predictions resulted in velocity components Ux = 1.08 m/s, Uy = 0.43 m/s, and static pressure p = 4.57 Pa. For NACA 6412, the corresponding mean absolute errors are Ux = 0.81 m/s, Uy = 0.59 m/s, and p = 7.5 Pa. Qualitative agreement was observed between PIV measurements, DeepCFD, and CFD. Results are promising that artificial intelligence has the potential for real-time fluid flow optimization of NACA airfoils in the future. The main goal was not just to train a network specifically for airfoils, but also for variant shapes. Airfoils are used since they are highly sophisticated in fluid dynamics and experimental data was available.

The author introduced models of linear logic known as ''Interaction Graphs'' which generalise Girard's various geometry of interaction constructions. In this work, we establish how these models essentially rely on a deep connection between zeta functions and the execution of programs, expressed as a cocycle. This is first shown in the simple case of graphs, before begin lifted to dynamical systems. Focussing on probabilistic models, we then explain how the notion of graphings used in Interaction Graphs captures a natural class of sub-Markov processes. We then extend the realisability constructions and the notion of zeta function to provide a realisability model of second-order linear logic over the set of all (discrete-time) sub-Markov processes.

Lab Automation facilitates high-throughput processes and improves reproducibility and efficiency while removing human action, primary source of contaminating particles. Handling poses a risk of contamination due to close contact with the objects. We propose a novel gripper (CrocoGrip) relying on compliant mechanisms to reduce the amount of contaminating particles generated by the gripper rather than preventing their emission, the latter being the common approach in current grippers. Our novel gripper is actuated by linear solenoids and purely relies on deformation for its motion. As a result, abrasive behavior and, therefore, the generation of particles is reduced without the need for additional sealing. We experimentally proved that only particles smaller than 3.0µm are emitted by the gripper, with a large proportion of the particles being generated by the actuation. The CrocoGrip fulfills the demands of ISO14644 class 5. The gripping relies on the deformation energy of the compliant mechanism, making the gripping energy-efficient and safe. The maximum gripping force achieved by the CrocoGrip was 5.5N. Because the force transmitted to the handling object depends on the design of the gripping jaws, which are interchangeable, the force can be reduced for more sensible handling objects. Using three different sets of jaws, CrocoGrip was able to handle a microplate in SBS-standard, a 50mL Falcon tube, and a Ø60mm Petri dish using a robotic arm. Due to the monolithic design of the CrocoGrip and, as a result, the need for few components, we achieve a simplicity of design, making cleaning, sterilization and maintenance easy, even for nonexperts. The CrocoGrip exploits the advantages of compliant mechanisms, especially for applications requiring clean-room environments. This approach of compliant-mechanism-based grippers enables an increase in the cleanliness of handling processes without an increase in system complexity of the gripper to facilitate the lab automation of highly sensible processes, such as in tissue engineering.

This study aims to evaluate the clinical outcome of stereotactic radiosurgery as the sole treatment for brain metastases and to assess prognostic factors influencing survival. A total of 108 consecutive patients with 213 metastases were retrospectively analyzed. Treatment was determined with close-meshed MRI follow-up. Various prognostic factors were assessed, and several prognostic indices were compared regarding their reliability to estimate overall survival. Median overall survival was 15 months; one-year overall survival was 50.5%. Both one- and two-year local controls were 90.9%. The rate of new metastases after SRS was 49.1%. Multivariate analysis of prognostic factors revealed that the presence of extracranial metastases, male sex, lower KPI, and progressive extracranial disease were significant risk factors for decreased survival. Of all evaluated prognostic indices, the Basic Score for Brain Metastases (BSBMs) showed the best correlation with overall survival. A substantial survival advantage was found for female patients after SRS when compared to male patients (18 versus 9 months, p = 0.003). SRS of brain metastasis is a safe and effective treatment option when frequent monitoring for new metastases with MRI is performed. Common prognostic scores lack reliable estimation of survival times. Female sex should be considered as an additional independent positive prognostic factor influencing survival.

Abstract Background This paper details the results of an evaluation of the level of consensus amongst clinicians on the use of ataluren in both ambulatory and non-ambulatory patients with nonsense mutation Duchenne muscular dystrophy (nmDMD). The consensus was derived using a modified Delphi methodology that involved an exploration phase and then an evaluation phase. Methods The exploration phase involved 90-minute virtual 1:1 interviews of 12 paediatric neurologists who cared for 30–120 DMD patients each and had patient contact every one or two weeks. The respondents managed one to ten nmDMD patients taking ataluren. The Discussion Guide for the interviews can be viewed as Appendix A. Following the exploration phase interviews, the interview transcripts were analysed by an independent party to identify common themes, views and opinions and developed 43 draft statements that the Steering Group (authors) reviewed, refined and endorsed a final list of 42 statements. Details of the recruitment of participants for the exploration and evaluation phases can be found under the Methods section. Results A consensus was agreed (> 66% of respondents agreeing) for 41 of the 42 statements using results from a consensus survey of healthcare professionals (n = 20) experienced in the treatment of nmDMD. Conclusions The statements with a high consensus suggest that treatment with ataluren should be initiated as soon as possible to delay disease progression and allow patients to remain ambulatory for as long as possible. Ataluren is indicated for the treatment of Duchenne muscular dystrophy that results from a nonsense mutation in the dystrophin gene, in ambulatory patients aged 2 years and older (see Summary of Product Characteristics for each country)

Abstract Dysfunctional Ca2+ signaling affects the myocardial systole and diastole, may trigger arrhythmia and cause transcriptomic and proteomic modifications in heart failure. Thus, synchronous real-time measurement of Ca2+ and force is essential to investigate the relationship between contractility and Ca2+ signaling and the alteration of excitation-contraction coupling (ECC) in human failing myocardium. Here, we present a method for synchronized acquisition of intracellular Ca2+ and contraction force in long-term cultivated slices of human failing myocardium. Synchronous time series of contraction force and intracellular Ca2+ were used to calculate force-calcium loops and to analyze the dynamic alterations of ECC in response to various pacing frequencies, post-pause potentiation, high mechanical preload and pharmacological interventions in human failing myocardium. We provide an approach to simultaneously and repeatedly investigate alterations of contractility and Ca2+ signals in long-term cultured myocardium, which will allow detecting the effects of electrophysiological or pharmacological interventions on human myocardial ECC.

Neural network-based nonlinear system identification is crucial for various multi-step ahead prediction tasks, including model predictive control and digital twins. These applications demand models that are not only accurate but also efficient in training and deployment. While current state-of-the-art neural network-based methods can identify accurate models, they often become prohibitively slow when scaled to achieve high accuracy, limiting their use in resource-constrained or time-critical applications. We propose FranSys, a Fast recurrent neural network-based method for multi-step ahead prediction in non-autoregressive System Identification. FranSys comprises three key innovations: 1) the first non-autoregressive RNN model structure for multi-step ahead prediction that enables much faster training and inference compared to autoregressive RNNs by separating state estimation and prediction into two specialized sub-models, 2) a state distribution alignment training technique that enhances generalizability and 3) a prediction horizon scheduling method that accelerates training by progressively increasing the prediction horizon. We evaluate FranSys on three publicly available benchmark datasets representing diverse systems, comparing its speed and accuracy against state-of-the-art RNN-based multi-step ahead prediction methods. The evaluation includes various prediction horizons, model sizes, and hyperparameter optimization settings, using both our own implementations and those from related work. Results demonstrate that FranSys is 10 to 100 times faster in training and inference with the same and often higher accuracy on test data than state-of-the-art RNN-based multi-step ahead prediction methods, particularly with long prediction horizons. This substantial speed improvement enables the application of larger neural network-based models with longer prediction horizons on resource-constrained systems in time-critical tasks, such as model predictive control and online learning of digital twins. The code of FranSys is publicly available.

Harmonization in the analytical framework is needed to detect, define and further categorize plastics released into the environment. In the range of particles smaller than 200 μm, hydrocyclones (HCs) have proven their capacity in removing microplastics efficiently by offering technical advantages at low operational costs. This publication aims to expand scientific knowledge by introducing four commercially available, low-priced microplastics to a pilot-scale HC setting. The physicochemical characteristics of particles as well as the separation efficiency of the test rig were investigated in depth. Particles with a density of >1000 kg/m3 passed the primary vortex and were discharged into the underflow, allowing us to employ standard mode operation. Particles with a density of 3 entered the secondary vortex and were removed through the overflow. As expected, separation efficiencies were found to be higher for particles revealing a greater density difference when compared with the mobile phase water. Furthermore, an increase in the inlet volume flow revealed significant positive impacts on the separation efficiency for three plastics to a certain threshold. Data on standard and reverse mode operations presented in this publication can lay out an important source for the harmonization and standardization of future HC research, with the goal of overcoming plastic pollution by developing economically competitive separation processes.

We propose a method to estimate information loss when conducting histogram binning and principal component analysis (PCA) sequentially, as usually done in practice for fleet analytics. Coarser-grained histogram binning results in less data volume, fewer dimensions, but more information loss. Considering fewer principal components (PCs) results in fewer data dimensions but increased information loss. Although information loss with each step is well understood, little guidance exists on the overall information loss when conducting both steps sequentially. We use Monte Carlo simulations to regress information loss on the number of bins and PCs, given few parameters of a dataset related to its scale and correlation structure. A sensitivity study shows that information loss can be approximated well given sufficiently large datasets. Using the number of bins, PCs, and two correlation measures, we derive an empirical loss model with high accuracy. Furthermore, we demonstrate the benefits of estimating information losses and the representativeness of total loss in evaluating the accuracy of k-means clustering for a real-world customer fleet dataset. For preprocessing sensor data which are aggregated from sufficient number of samples, continuously distributed, and can be represented by Beta-distributions, we recommend not to coarsen the histogram binning before PCA.

A polymicrobial biofilm model of Komagataeibacter hansenii and Pseudomonas aeruginosa was developed to understand whether a pre-existing matrix affects the ability of another species to build a biofilm. P. aeruginosa was inoculated onto the preformed K. hansenii biofilm consisting of a cellulose matrix. P. aeruginosa PAO1 colonized and infiltrated the K. hansenii bacterial cellulose biofilm (BC), as indicated by the presence of cells at 19 μm depth in the translucent hydrogel matrix. Bacterial cell density increased along the imaged depth of the biofilm (17-19 μm). On day 5, the average bacterial count across sections was 67 ± 4 % P. aeruginosa PAO1 and 33 ± 6 % K. hansenii. Biophysical characterization of the biofilm indicated that colonization by P. aeruginosa modified the biophysical properties of the BC matrix, which inlcuded increased density, heterogeneity, degradation temperature and thermal stability, and reduced crystallinity, swelling ability and moisture content. This further indicates colonization of the biofilm by P. aeruginosa. While eDNA fibres - a key viscoelastic component of P. aeruginosa biofilm - were present on the surface of the co-cultured biofilm on day 1, their abundance decreased over time, and by day 5, no eDNA was observed, either on the surface or within the matrix. P. aeruginosa-colonized biofilm devoid of eDNA retained its mechanical properties. The observations demonstrate that a pre-existing biofilm scaffold of K. hansenii inhibits P. aeruginosa PAO1 eDNA production and suggest that eDNA production is a response by P. aeruginosa to the viscoelastic properties of its environment.

BackgroundAnti-IgLON5 disease is a rare chronic autoimmune disorder characterized by IgLON5 autoantibodies predominantly of the IgG4 subclass. Distinct pathogenic effects were described for anti-IgLON5 IgG1 and IgG4, however, with uncertain clinical relevance.MethodsIgLON5-specific IgG1-4 levels were measured in 46 sera and 20 cerebrospinal fluid (CSF) samples from 13 HLA-subtyped anti-IgLON5 disease patients (six females, seven males) using flow cytometry. Intervals between two consecutive serum or CSF samplings (31 and 10 intervals, respectively) were categorized with regard to the immunomodulatory treatment active at the end of the interval, changes of anti-IgLON5 IgG1 and IgG4 levels, and disease severity. Intrathecal anti-IgLON5 IgG4 synthesis (IS) was assessed using a quantitative method.ResultsThe median age at onset was 66 years (range: 54–75), disease duration 10 years (range: 15–156 months), and follow-up 25 months (range: 0–83). IgLON5-specific IgG4 predominance was observed in 38 of 46 (83%) serum and 11 of 20 (55%) CSF samples. Anti-IgLON5 IgG4 levels prior clinical improvement in CSF but not serum were significantly lower than in those prior stable/progressive disease. Compared to IgLON5 IgG4 levels in serum, CSF levels in HLA-DRB1*10:01 carriers were significantly higher than in non-carriers. Indeed, IgLON5-specific IgG4 IS was demonstrated not only in four of five HLA-DRB1*10:01 carriers but also in one non-carrier. Immunotherapy was associated with decreased anti-IgGLON5 IgG serum levels. In CSF, lower anti-IgLON5 IgG was associated with immunosuppressive treatments used in combination, that is, corticosteroids and/or azathioprine plus intravenous immunoglobulins or rituximab.ConclusionOur findings might indicate that CSF IgLON5-specific IgG4 is frequently produced intrathecally, especially in HLA-DRB1*10:01 carriers. Intrathecally produced IgG4 may be clinically relevant. While many immunotherapies reduce serum IgLON5 IgG levels, more intense immunotherapies induce clinical improvement and may be able to target intrathecally produced anti-IgLON5 IgG. Further studies need to confirm whether anti-IgLON5 IgG4 IS is a suitable prognostic and predictive biomarker in anti-IgLON5 disease.

Tannins, which can be extracted from certain tree barks, are polyphenolic compounds, that gain a lot of interest in industry, as they can be utilized to replace petroleum-based chemicals. Currently, they are used in the leather industry, as flocculants or as adhesives. To further increase their potential, an increase in purity is necessary. To do so, various methods are available, in this study we present a 2-step approach consisting of (i) depolymerization and cleavage of glycosidic bonds and (ii) separation of phenolic-rich and phenolic-poor fractions. Commercially available mimosa tannin is treated with various concentrations of hydrochloric and sulfuric acid (0.4 M and 0.8 M) and separated with chromatographic resin. Resulting fractions are characterized by UV/Vis spectroscopy for phenolic content and antioxidant activity, infrared spectroscopy, NMR, HPLC-DAD and GPC to determine their composition and molecular size as well as their reactivity towards formaldehyde. It was observed that mild acidic cleavage with 0.4 M HCl results in more phenol-rich material (TPC 1124 µgGAE mg−1) while 0.8 M sulfuric acid leads to tannin fractions with higher antioxidative properties. Amberlite resin yields fractions with higher reactivity towards formaldehyde (gel time below 60 s compared to over 90 originally) while the Sephadex fractions present the highest values for total phenolic content.

Simultaneous gas phase temperature and water vapor concentration measurement are important to understand reacting flows such as combustion or gas reforming processes. Here, coherent anti-Stokes Raman scattering (CARS) offers the possibility for non-intrusive measurements with a high temporal and spatial resolution. Therefore, this work demonstrates the simultaneous measurement of temperature and relative water vapor–nitrogen concentrations by using dual-pump vibrational coherent anti-Stokes Raman scattering (DPVCARS). A calibration procedure is developed for a temperature range of 473 K to 673 K and a water vapor concentration of 24% to 46% at ambient pressure. This setup is tested with 500 CARS single pulse spectra taken in a gas cell at a known temperature and concentration. Based on these results, information about precision and accuracy can be delivered.

Abstract Expression of CYP3A5 protein is a basal and acquired resistance mechanism of pancreatic ductal adenocarcinoma cells conferring protection against the CYP3A and CYP2C8 substrate paclitaxel through metabolic degradation. Inhibition of CYP3A isozymes restores the cells sensitivity to paclitaxel. The combination of gemcitabine and nab‐paclitaxel is an established regimen for the treatment of metastasized or locally advanced inoperable pancreatic cancer. Cobicistat is a CYP3A inhibitor developed for the pharmacoenhancement of protease inhibitors. The addition of cobicistat to gemcitabine and nab‐paclitaxel may increase the antitumor effect. We will conduct a phase I dose escalation trial with a classical 3 + 3 design to investigate the safety, tolerability, and pharmacokinetics (PKs) of gemcitabine, nab‐paclitaxel, and cobicistat. Although the doses of gemcitabine (1000 mg/m2) and cobicistat (150 mg) are fixed, three dose levels of nab‐paclitaxel (75, 100, and 125 mg/m2) will be explored to account for a potential PK drug interaction. After the dose escalation phase, we will set the recommended dose for expansion (RDE) and treat up to nine patients in an expansion part of the trial. The trial is registered under the following identifiers EudraCT‐Nr. 2019‐001439‐29, drks.de: DRKS00029409, and ct.gov: NCT05494866. Overcoming resistance to paclitaxel by CYP3A5 inhibition may lead to an increased efficacy of the gemcitabine and nab‐paclitaxel regimen. Safety, efficacy, PK, and RDE data need to be acquired before investigating this combination in a large‐scale clinical study.

Abstract The extracellular matrix of bacterial biofilms consists of diverse components including polysaccharides, proteins and DNA. Extracellular RNA (eRNA) can also be present, contributing to the structural integrity of biofilms. However, technical difficulties related to the low stability of RNA make it difficult to understand the precise roles of eRNA in biofilms. Here, we show that eRNA associates with extracellular DNA (eDNA) to form matrix fibres in Pseudomonas aeruginosa biofilms, and the eRNA is enriched in certain bacterial RNA transcripts. Degradation of eRNA associated with eDNA led to a loss of eDNA fibres and biofilm viscoelasticity. Compared with planktonic and biofilm cells, the biofilm matrix was enriched in specific mRNA transcripts, including lasB (encoding elastase). The mRNA transcripts colocalised with eDNA fibres in the biofilm matrix, as shown by single molecule inexpensive FISH microscopy (smiFISH). The lasB mRNA was also observed in eDNA fibres in a clinical sputum sample positive for P. aeruginosa. Thus, our results indicate that the interaction of specific mRNAs with eDNA facilitates the formation of viscoelastic networks in the matrix of Pseudomonas aeruginosa biofilms.

Abstract Background Transjugular intrahepatic portosystemic shunt (TIPS) is a well-validated treatment option for clinically significant portal hypertension (CSPH) in the context of liver cirrhosis. Its high efficacy and safety in the management of treatment-refractory ascites and variceal bleeding have been extensively proven. Contraindications for TIPS include severe right heart failure, hepatic encephalopathy, and sepsis. However, the role of liver malignancy in TIPS is debatable. Mostly, primary liver malignancies such as hepatocellular carcinoma (HCC) emerge from advanced liver diseases. Coexisting portal hypertension in HCC often results in limited treatment options and a poor prognosis. Summary Previous studies have shown that TIPS implantation in patients with HCC is technically feasible and is usually not associated with major adverse events. Furthermore, TIPS may help in bridging the time to liver transplantation in early HCC and allow for locoregional treatment in advanced HCC. However, several studies suggest that seeding tumour cells to the lungs by TIPS placement might worsen the prognosis. Conclusions TIPS placement in patients with coexisting liver malignancy remains a case-by-case decision, and there is no profound evidence allowing general recommendations. This review aims to provide a state-of-the-art overview of the potential risks and benefits of TIPS placement in patients with liver malignancies.

Abstract Objectives It is uncertain, if omitting post-dilatation and stent oversizing (stent optimization) is safe and may decrease the risk for distal thrombus embolization (DTE) in STEMI patients with large thrombus burden (LTB). Background In patients with ST-segment elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (pPCI) with stenting, (DTE) and flow deterioration are common and increase infarct size leading to worse outcomes. Methods From a prospective registry, 74 consecutive STEMI patients with LTB undergoing pPCI with stenting and intentionally deferred stent optimization were analyzed. Imaging data and outcomes up to 2 years follow-up were analyzed. Results Overall, 74 patients (18% females) underwent deferred stent optimization. Direct stenting was performed in 13 (18%) patients. No major complications occurred during pPCI. Staged stent optimization was performed after a median of 4 (interquartile range (IQR) 3; 7) days. On optical coherence tomography, under-expansion and residual thrombus were present in 59 (80%) and 27 (36%) cases, respectively. During deferred stent optimization, we encountered no case of flow deterioration (slow or no-reflow) or side branch occlusion. Minimal lumen area (mm2) and stent expansion (%) were corrected from 4.87±1.86mm to 6.82±2.36mm (p

Abstract Global cooling has been proposed as a driver of the Great Ordovician Biodiversification Event, the largest radiation of Phanerozoic marine animal Life. Yet, mechanistic understanding of the underlying pathways is lacking and other possible causes are debated. Here we couple a global climate model with a macroecological model to reconstruct global biodiversity patterns during the Ordovician. In our simulations, an inverted latitudinal biodiversity gradient characterizes the late Cambrian and Early Ordovician when climate was much warmer than today. During the Mid-Late Ordovician, climate cooling simultaneously permits the development of a modern latitudinal biodiversity gradient and an increase in global biodiversity. This increase is a consequence of the ecophysiological limitations to marine Life and is robust to uncertainties in both proxy-derived temperature reconstructions and organism physiology. First-order model-data agreement suggests that the most conspicuous rise in biodiversity over Earth’s history – the Great Ordovician Biodiversification Event – was primarily driven by global cooling.

Abstract Background Alcohol is a good and environment-friendly fuel that can be microbially produced, capable of eliminating many of the limitations of the present-day fossil fuels. However, the inherent toxic nature of alcohols to the microbial cells leads to end-product inhibition that limits large-scale alcohol production by fermentation. Fundamental knowledge about the stress responses of microorganisms to alcohols would greatly facilitate to improve the microbial alcohol tolerance. The current study elucidates and compares the changes in the membrane proteome of Escherichia coli in response to a range of alcohols. Results Although alcohol toxicity increased exponentially with alcohol chain length (2–6 carbon), similar stress responses were observed in the inner and outer membrane proteome of E. coli in the presence of 2-, 4- and 6-carbon alcohols at the MIC50. This pertains to: (1) increased levels of inner membrane transporters for uptake of energy-producing metabolites, (2) reduced levels of non-essential proteins, associated with anaerobic, carbon starvation and osmotic stress, for energy conservation, (3) increased levels of murein degrading enzymes (MltA, EmtA, MliC and DigH) promoting cell elongation and 4) reduced levels of most outer membrane β-barrel proteins (LptD, FadL, LamB, TolC and BamA). Major outer membrane β-barrel protein OmpC, which is known to contribute to ethanol tolerance and membrane integrity, was notably reduced by alcohol stress. While LPS is important for OmpC trimerisation, LPS release by EDTA did not lower OmpC levels. This suggests that LPS release, which is reported under alcohol stress, does not contribute to the reduced levels of OmpC in the presence of alcohol. Conclusions Since alcohol primarily targets the integrity of the membrane, maintenance of outer membrane OmpC levels in the presence of alcohol might help in the survival of E. coli to higher alcohol concentrations. The study provides important information about the membrane protein responses of E. coli to a range of alcohols, which can be used to develop targeted strategies for increased microbial alcohol tolerance and hence bioalcohol production.

Contemporary art consumption is predominantly online, driven by algorithmic recommendation systems that dictate artwork visibility. Despite not being designed for curation, these algorithms’ machinic ways of seeing play a pivotal role in shaping visual culture, influencing artistic creation, visibility, and associated social and financial benefits. The Algorithmic Pedestal was a gallery, practice-based research project that reported gallerygoers’ perceptions of a human’s curation and curation achieved by Instagram’s algorithm. This paper presents a technical analysis of the same exhibit using computer vision code, offering insights into machines’ perception of visual art. The computer vision code assigned values on various metrics to each image, allowing statistical comparisons to identify differences between the collections of images selected by the human and the algorithmic system. The analysis reveals statistically significant differences between the exhibited images and the broader Metropolitan Museum of Art digital collection. However, the analysis found minimal distinctions between human-curated and Instagram-curated images. This study contributes insights into the perceived value of the curation process, shedding light on how audiences perceive artworks differently from machines using computer vision.

Background: Intracellular tracking is commonly used in trafficking research. Until today, the respective techniques have remained complex, and complicated, mostly transgenic target protein changes are necessary, often requiring expensive equipment and expert knowledge. Methods: We present a novel method, which we term “cell-sonar”, that enables the user to track expression changes of specific protein markers that serve as points of interaction. Our study provides comparable analyses of expression changes of these marker proteins by in-cell Western analyses in two otherwise isogenic cell lines that only differ in the overexpression of the tracked target protein. Using the overexpressed human adult muscle-type nicotinic acetylcholine receptor as an example, we demonstrate that cell-sonar can cover multiple intracellular compartments such as the endoplasmic reticulum, the pathway between it and the Golgi apparatus, and the endocytic pathway. Results: We provide evidence for receptor maturation in the Golgi and storage in recycling endosomes, rather than the fate of increased insertion into the plasma membrane. Additionally, we demonstrate with the implementation of nicotine that the receptor’s destiny is exasperated up to secondary degradation. Conclusions: Cell-sonar is an affordable, easy-to-implement, and cheap method that can be adapted to a broad variety of proteins and cellular pathways of interest to researchers.

Cyclones are pivotal in mechanical process engineering and crucial in the complex field of separation technology. Their robustness and compact spatial requirements render them universally applicable and versatile across various industrial domains. Depending on the utilized fluid and field of application, both gas-based cyclones and hydrocyclones (HCs) are well established. Regarding HC design, enduring elongated flat cones have seen minimal alterations in shape and structure since their introduction over more than a hundred years ago. Experimental investigations regarding unconventional cone designs within scientific studies remain the exception. Therefore, this study focuses on alternative geometric configurations of the separation chambers and highlights their impact on separation and energy efficiency. To achieve this objective, different geometric shapes are investigated and retrofitted into HCs. The geometric foundation is derived from upcycled glass bottles. The repurposed bottles with a volume of 750 mL are used in conjunction with an inlet part, following the established Rietema design. Experimental tests are conducted with dilute phase separation, using 0.1–200 µm test particles in water. Comparisons between a bottle-based HC and a conventional Rietema design were conducted, establishing a benchmark against the standard. The findings revealed a noticeable correlation between separation efficiency and cone geometry. Conical designs demonstrated enhanced separation, particularly at lower volume flows. At the highest volume flow of 75 L min−1, the best performing bottle cyclones showed separation efficiencies of 78.5%, 78.4% and 77.9% and therefore are in a competitive range with 78.0% efficiency, achieved using the commercial Rietema design. Minimal disparities in cut sizes were observed in terms of separation grade efficiency among the models tested. Variations in separation efficiency and fractional efficiency curves indicated nuanced differences in classification efficiency.

Abstract Background Real-world data on treatment patterns/outcomes for metastatic pancreatic cancer (mPAC) are limited. This study aims to assess real-world treatment patterns, survival outcomes, and prognostic/predictive factors in patients with mPAC. Methods Retrospective, observational, chart-review involving medical oncologists and gastroenterologists from five European countries. Physicians reported information on disease and patient characteristics, diagnosis, and treatment for patients diagnosed with mPAC from January-October 2016. Outcomes included median progression-free survival (mPFS), median overall survival (mOS), and the impact of baseline performance status on survival. Univariate/multivariate regression analyses were undertaken to identify prognostic/predictive factors. Results Three hundred four physicians and 3432 patients were included. First-line therapies included modified (m)FOLFIRINOX (28.4%), gemcitabine + nab-paclitaxel (28.0%), and gemcitabine monotherapy (23.0%). Frequent second-line therapies were gemcitabine monotherapy (25.0%), fluorouracil (5-FU) + oxaliplatin (21.8%), and gemcitabine + nab-paclitaxel (16.7%). Most frequent first- to second-line treatment sequences were gemcitabine + nab-paclitaxel followed by fluoropyrimidine combinations. Longest unadjusted estimated mOS was observed with (m)FOLFIRINOX followed by gemcitabine-based combinations (19.1 months). Multivariate analysis identified significant prognostic/predictive factors for OS and PFS including performance status and carbohydrate antigen 19–9 (CA 19–9) levels. Conclusions Treatment and treatment sequences were generally in accordance with guidelines at the time of the study. Identification of prognostic/predictive factors for survival may help inform the individualised management of mPAC patients in the future.

Abstract Climatic and edaphic effects are increasingly being discussed in the context of biodiversity-ecosystem functioning. Here we use data from West African semi-arid tree savannas and contrasting climatic conditions (lower vs. higher mean annual precipitation-MAP and mean annual temperature-MAT) to (1) determine how climate modulates the effects of species richness on aboveground carbon (AGC); (2) explore how species richness and AGC relate with soil variables in these contrasting climatic conditions; and (3) assess how climate and soil influence directly, and/or indirectly AGC through species richness and stand structural attributes such as tree density and size variation. We find that greater species richness is generally associated with higher AGC, but more strongly in areas with higher MAP, which also have greater stem density. There is a climate-related influence of soils on AGC, which decreases from lower to higher MAP conditions. Variance partitioning analyses and structural equation modelling show that, across all sites, MAP, relative to soils, has smaller effect on AGC, mediated by stand structural attributes whereas soil texture and fertility explain 14% of variations in AGC and influence AGC directly and indirectly via species richness and stand structural attributes. Our results highlight coordinated effects of climate and soils on AGC, which operated primarily via the mediation role of species diversity and stand structures.

Abstract Key message We provide a set of allometric models for wild cherry trees (Prunus avium L.) established in agroforestry systems. A total of 70 trees in southwestern Germany were surveyed using terrestrial laser scanning and analysed using quantitative structure models. The derived allometric models provide a stable base for biomass estimation in comparable agroforestry systems. Our biomass model, based on volume estimates converted to biomass, shows no significant differences to a previous study in the same region on the same species, although it was conducted on agroforestry trees under a different management regime. Context Wild cherry (Prunus avium L.) is a common tree species in agroforestry systems (AFS). Utilised for either fruit production or for high-value timber production, it is a highly relevant species, yet even basic allometric models are lacking. Aims The aim of this study was to develop a set of allometric models for wild cherry trees in AFS. Within this context, we present an innovative non-destructive approach to estimate bark and wood volume separately by applying bark thickness models to 3D models of trees. To assess model applicability to different AFS, we compared our allometric model for above-ground biomass with a previous biomass model for wild cherry trees under different management in the same region. Methods Wild cherry trees (n = 70) located within AFS in southern Germany were scanned with a terrestrial laser scanner. Quantitative structure models were used to derive tree dimensions and above-ground volume per tree. Using additional auxiliary data, the target variables were derived, and corresponding allometric models were fitted. Results The allometric models estimating above-ground volume, oven-dry biomass, carbon content and nutrient content based on diameter at breast height (DBH) showed excellent fits (R 2 adj ≥ 0.97). The comparisons with a similar study conducted in the same region suggested that management practices such as pruning have only a minor influence on the relationship between DBH and above-ground tree biomass. The nutrient content in the trees decreased in the order Ca > N > K > Mg > P. Conclusions The derived allometric models provide valuable information on this important agroforestry tree species. Our findings can both inform management practices in AFS and advance ecological understanding of these systems. Future research should focus on developing allometric models for other tree species relevant to AFS.

Introduction: The plant-based alkaloid muscimol is a potent agonist of inhibitory GABAA-neurotransmitter receptors. GABAA receptors are a heterogeneous family of pentameric complexes, with 5 out of 19 subunits assembling around the central anion pore. Muscimol is considered to bind to all receptor subtypes at the orthosteric drug binding site at the β+/α− interface. Recently, we observed that the antipsychotic drugs clozapine (CLZ), loxapine (LOX) and chlorpromazine (CPZ) although exerting functional inhibition on multiple GABAA receptor subtypes showed diverging results in displacing 3H-muscimol. While a complete displacement could be observed in hippocampal membranes by bicuculline (BIC), and no displacement with CPZ, the compounds CLZ and LOX competed partially. Non-sigmoidal, complex dose response curves were indicative of multiple sites. In the current study we now aimed to investigate more extensively this heterogeneity of bicuculline sensitive muscimol sites in rat brain.Methods: We tested membranes from four different brain regions (hippocampus, cerebellum, thalamus and striatum) and selected recombinantly expressed subunit combinations with displacement assays. 3H-muscimol displacement was tested with BIC, LOX, CLZ and CPZ. In silico ligand structural analysis and computational docking was performed.Results: We observed a unique pharmacology of each tested compound in the studied brain regions. Combining two of the tested ligands suggests that in striatum all CLZ sites are contained in the pool of LOX sites, while the CPZ sites may in part be non-overlapping with LOX sites. Experiments on recombinantly expressed receptors indicate, that BIC can displace 3H-muscimol from all tested receptors, while LOX and CLZ display different and variable competition indicative of multiple sites. Molecular docking produced structural correlates of the observed diversity of muscimol sites on the basis of bicuculline bound experimental structures.Discussion: These findings indicate that 3H-muscimol binding sites in rat brain are heterogeneous, with different populations of receptors, which are CPZ, LOX or CLZ sensitive or insensitive. These binding sites show a varying distribution in different rat brain regions. Molecular docking suggests that the so-called loop F region of α subunits drives the observed differences.

Introduction Task-shifting from primary care physicians (PCPs) to nurses is one option to better and more efficiently meet the needs of the population in primary care and to overcome PCP shortages. This protocol outlines an overview of systematic reviews to assess the effects of delegation or substitution by nurses of PCPs’ activities regarding clinical, patient-relevant, professional and health services-related outcomes.Methods and analysis We will conduct a systematic literature search for secondary literature in PubMed/MEDLINE, EMBASE, CINAHL and Cochrane databases. Systematic reviews, meta-analyses and Health Technology Assessments in German and English comprising randomised controlled trials and prospective controlled trials will be considered for inclusion. Search terms will include Medical Subject Headings combined with free text words. At least one-third of abstracts and full-text articles are reviewed by two independent reviewers. Methodological quality will be assessed using the Overview Quality Assessment Questionnaire. We will only consider reviews if they include controlled trials, if the profession that substituted or delegated tasks was a nurse, if the profession of the control was a PCP, if the assessed intervention was the same in the intervention and control group and if the Overview Quality Assessment Questionnaire score is ≥5. The corrected covered area will be calculated to describe the degree of overlap of studies in the reviews included in the study. We will report the overview according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses.Ethics and dissemination The overview of secondary literature does not require the approval of an Ethics Committee and will be published in a peer-reviewed journal.PROSPERO registration number CRD42020183327.

IntroductionThe understanding of health-related information is essential for making informed decisions. However, providing health information in an understandable format for everyone is challenging due to differences in consumers’ health status, disease knowledge, skills, and preferences. Tailoring health information to individual needs can improve comprehension and increase health literacy.ObjectiveThe aim of our research was to analyze the extent to which consumers can customize consumer health information materials (CHIMs) for type-2 diabetes mellitus through various media types.MethodsWe conducted a comprehensive search for various CHIMs across various media types, such as websites, apps, videos, and printed or printable forms. A representative sample of CHIMs was obtained for analysis through blocked randomization across the various media types. We conducted a quantitative content analysis to determine the frequency of user-centered customization options. Cross-comparisons were made to identify trends and variations in modifiable features among the media.ResultsIn our representative sample of 114 CHIMs, we identified a total of 24 modifiable features, which we grouped into five main categories: (i) language, (ii) text, (iii) audiovisual, (iv) presentation, and (v) medical content. Videos offered the most customization opportunities (95%), while 47% of websites and 26% of apps did not allow users to tailor health information. None of the printed or printable materials provided the option to customize the information. Overall, 65% of analyzed CHIMs did not allow users to tailor health information according to their needs.ConclusionOur results show that CHIMs for type-2 diabetes mellitus could be significantly improved by providing more customization options for users. Further research is needed to investigate the effectiveness and usability of these options to enhance the development and appropriate provision of modifiable features in health information.

Trees are able to adapt to changing environmental conditions through modifications in their crown morphology and branch architecture. In light-limited environments, tree structures are mainly optimised to increase light interception. This might have side effects on other properties such as the hydraulic system that determines water conduction and tree reactions to drought events. Given the increasing exposure of forest ecosystems to drought stress it is thus crucial to investigate possible correlations between the crown morphology and branch architecture and drought responses. Our study aimed to compare different crown archetypes and branch characteristics of European beech (Fagus sylvatica L.) in their reaction to drought, which was assessed by measuring tree-ring widths of increment cores sampled at breast height. Using Terrestrial Laser Scanning and Quantitative Structure Models, we explored various crown morphological and branch architectural characteristics of 67 beech trees and identified three species-specific crown archetypes. The crown archetypes and branch variables were contrasted with growth responses using linear mixed models. While productivity levels differed, the negative impact of drought stress on radial growth was consistent across all crown archetypes. Nevertheless, certain branch architectural variables were important predictors for radial growth in drought situations. Specifically, long water conduction paths and many branching nodes along those paths were positively influencing growth. Our results indicate that trees showing these characteristics might have a competitive edge regarding drought-affected radial growth compared to others. They could be promoted through thinning measures, which would allow improving the adaption of existing beech forests in situ to climate change and drought stress.

Desmoplastic small round blue cell tumor (DSRCT) is a highly aggressive fatal sarcoma without evidence-based therapeutic guidelines. We present here seven patients with DSRCT including immunohistochemistry combined with fluorescence in situ hybridization (FISH), next generation sequencing (NGS, n = 6) as well as OncoScan array (n = 3) analyses and show consecutive therapeutic approaches. All seven DSRCT patients presented with an extended abdominal mass; median age at diagnosis was 24.8 years. NGS analyses revealed five class 4 or 5 sequence variants. Remarkably, OncoScan and targeted analyses by FISH identified genomic gains of CCND1 in two cases. Cyclin D1 expression was present in all seven tumors as shown by immunohistochemical staining. Multimodal therapeutic concepts included systemic therapies, resection, and radiation. Six patients were treated as first-line therapy with conventional chemotherapy. All except one patient had a dismal therapy response. Subsequent therapy lines consisted of chemotherapeutic combinations followed by targeted therapies. Due to Cyclin D1 expression, the CDK4/6 inhibitor palbociclib was applied to four patients. The median therapy duration until disease progression in these patients was 4.5 months (range, 1.5–5 months). So, CCND1 genomic gain and Cyclin D1 expression are common features pointing to cell-cycle deregulation as a possible therapeutic target.

BackgroundAnti-vascular endothelial growth factor (VEGF) monoclonal antibodies (mAbs) are widely used for tumor treatment, including metastatic colorectal cancer (mCRC). So far, there are no biomarkers that reliably predict resistance to anti-VEGF mAbs like bevacizumab. A biomarker-guided strategy for early and accurate assessment of resistance could avoid the use of non-effective treatment and improve patient outcomes. We hypothesized that repeated analysis of multiple cytokines and angiogenic growth factors (CAFs) before and during treatment using machine learning could provide an accurate and earlier, i.e., 100 days before conventional radiologic staging, prediction of resistance to first-line mCRC treatment with FOLFOX plus bevacizumab.Patients and methods15 German and Austrian centers prospectively recruited 50 mCRC patients receiving FOLFOX plus bevacizumab as first-line treatment. Plasma samples were collected every two weeks until radiologic progression (RECIST 1.1) as determined by CT scans performed every 2 months. 102 pre-selected CAFs were centrally analyzed using a cytokine multiplex assay (Luminex, Myriad RBM).ResultsUsing random forests, we developed a predictive machine learning model that discriminated between the situations of "no progress within 100 days before radiological progress" and "progress within 100 days before radiological progress". We could further identify a combination of ten out of the 102 CAF markers, which fulfilled this task with 78.2% accuracy, 71.8% sensitivity, and 82.5% specificity.ConclusionsWe identified a CAF marker combination that indicates treatment resistance to FOLFOX plus bevacizumab in patients with mCRC within 100 days prior to radiologic progress.

BackgroundWe have developed a new clinical research approach for the quantification of cellular proliferation in human infants to address unanswered questions about tissue renewal and regeneration. The approach consists of oral 15N-thymidine administration to label cells in S-phase, followed by Multi-isotope Imaging Mass Spectrometry for detection of the incorporated label in cell nuclei. To establish the approach, we performed an observational study to examine uptake and elimination of 15N-thymidine. We compared at-home label administration with in-hospital administration in infants with tetralogy of Fallot, a form of congenital heart disease, and infants with heart failure.MethodsWe examined urine samples from 18 infants who received 15N-thymidine (50 mg/kg body weight) by mouth for five consecutive days. We used Isotope Ratio Mass Spectrometry to determine enrichment of 15N relative to 14N (%) in urine.Results/findings15N-thymidine dose administration produced periodic rises of 15N enrichment in urine. Infants with tetralogy of Fallot had a 3.2-fold increase and infants with heart failure had a 4.3-fold increase in mean peak 15N enrichment over baseline. The mean 15N enrichment was not statistically different between the two patient populations (p = 0.103). The time to peak 15N enrichment in tetralogy of Fallot infants was 6.3 ± 1 hr and in infants with heart failure 7.5 ± 2 hr (mean ± SEM). The duration of significant 15N enrichment after a dose was 18.5 ± 1.7 hr in tetralogy of Fallot and in heart failure 18.2 ± 1.8 hr (mean ± SEM). The time to peak enrichment and duration of enrichment were also not statistically different (p = 0.617 and p = 0.887).ConclusionsThe presented results support two conclusions of significance for future applications: (1) Demonstration that 15N-thymidine label administration at home is equivalent to in-hospital administration. (2) Two different types of heart disease show no differences in 15N-thymidine absorption and elimination. This enables the comparative analysis of cellular proliferation between different types of heart disease.

BackgroundIn resource-constrained settings, limited antibiotic options make treating carbapenem-resistant bacterial infections difficult for healthcare providers. This study aimed to assess carbapenemase expression in Gram-negative bacteria isolated from clinical samples in Jimma, Ethiopia.MethodsA cross-sectional study was conducted to assess carbapenemase expression in Gram-negative bacteria isolated from patients attending Jimma Medical Center. Totally, 846 Gram-negative bacteria were isolated and identified using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Phenotypic antibiotic resistance patterns were determined using the Kirby-Bauer disk diffusion method and Etest strips. Extended-spectrum β-lactamase phenotype was determined using MAST disks, and carbapenemases were characterized using multiplex polymerase chain reactions (PCR).ResultsAmong the isolates, 19% (157/846) showed phenotypic resistance to carbapenem antibiotics. PCR analysis revealed that at least one carbapenemase gene was detected in 69% (107/155) of these strains. The most frequently detected acquired genes were blaNDM in 35% (37/107), blaVIM in 24% (26/107), and blaKPC42 in 13% (14/107) of the isolates. Coexistence of two or more acquired genes was observed in 31% (33/107) of the isolates. The most common coexisting acquired genes were blaNDM + blaOXA-23, detected in 24% (8/33) of these isolates. No carbapenemase-encoding genes could be detected in 31% (48/155) of carbapenem-resistant isolates, with P. aeruginosa accounting for 85% (41/48) thereof.ConclusionThis study revealed high and incremental rates of carbapenem-resistant bacteria in clinical samples with various carbapenemase-encoding genes. This imposes a severe challenge to effective patient care in the context of already limited treatment options against Gram-negative bacterial infections in resource-constrained settings.

Abstract Sulfur intercalation of a carbon rich (63×63)R30∘ reconstruction on silicon carbide, also known as buffer layer, is reported. In a two‐zone furnace a sulfur rich precursor is heated and the gaseous species is transported for intercalation by an argon flow to the sample. Successful intercalation can be confirmed by X‐ray photoelectron spectroscopy and low‐energy electron diffraction. Angle‐resolved photoelectron spectroscopy reveals a p‐type doping of the intercalated samples. In some cases only partial intercalation appears with non‐intercalated sulfur on top of the remaining buffer layer areas. Further annealing of such samples leads to a migration of the non‐intercalated sulfur under the buffer layer areas, indicating that the sulfur bonded to the buffer layer constitutes a transition state.

Introduction: Regulatory T cells (Tregs) play a critical role in the maintenance of immune homeostasis but also protect tumors from immune-mediated growth control or rejection and pose a significant barrier to effective immunotherapy. Inhibition of MALT1 paracaspase activity can selectively reprogram immune-suppressive Tregs in the tumor microenvironment to adopt a proinflammatory fragile state, which offers an opportunity to impede tumor growth and enhance the efficacy of immune checkpoint therapy (ICT). Methods: We performed preclinical studies with the orally available allosteric MALT1 inhibitor (S)-mepazine as a single-agent and in combination with anti-programmed cell death protein 1 (PD-1) ICT to investigate its pharmacokinetic properties and antitumor effects in several murine tumor models as well as patient-derived organotypic tumor spheroids (PDOTS). Results: (S)-mepazine demonstrated significant antitumor effects and was synergistic with anti-PD-1 therapy in vivo and ex vivo but did not affect circulating Treg frequencies in healthy rats at effective doses. Pharmacokinetic profiling revealed favorable drug accumulation in tumors to concentrations that effectively blocked MALT1 activity, potentially explaining preferential effects on tumor-infiltrating over systemic Tregs. Conclusions: The MALT1 inhibitor (S)-mepazine showed single-agent anticancer activity and presents a promising opportunity for combination with PD-1 pathway-targeted ICT. Activity in syngeneic tumor models and human PDOTS was likely mediated by induction of tumor-associated Treg fragility. This translational study supports ongoing clinical investigations (ClinicalTrials.gov Identifier: NCT04859777) of MPT-0118, (S)-mepazine succinate, in patients with advanced or metastatic treatment-refractory solid tumors.