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La displasia septo-óptica es una afección neurológica congénita de etiología multifactorial caracterizada por agenesia del septum pellucidum, disgenesia del cuerpo calloso o ambas, hipoplasia del quiasma o nervios ópticos y disfunción hormonal con alteraciones hipofisiarias o hipotalámicas. Para hacer el diagnóstico se requieren dos de estos criterios y la resonancia magnética es el examen de elección. La mayoría de los casos se presentan con anomalías del desarrollo cortical en la forma conocida como displasia septo-óptica plus. Si bien las convulsiones y los trastornos del neurodesarrollo son las manifestaciones neurológicas dominantes, es una entidad muy heterogénea con múltiples hallazgos clínicos y radiológicos que se deben considerar. Se presenta el caso de un hombre de 35 años con antecedentes de traumatismo craneoencefálico en la infancia y remisión por epilepsia focal resistente al tratamiento asociada con déficit cognitivo. En la evaluación inicial, la tomografía computarizada de cráneo simple se apreció agenesia del septum pellucidum y disgenesia del cuerpo calloso. Se practicó una resonancia magnética cerebral que confirmó la agenesia del septum pellucidum y, también, reveló irregularidad y engrosamiento anómalo de la corteza cerebral en los lóbulos frontales y la región perisilviana; además, se encontró sustancia gris heterotópica en los lóbulos frontales y la región frontoinsular izquierda, leve ventriculomegalia supratentorial, apariencia atípica del rostrum del cuerpo calloso e hipoplasia del quiasma y los nervios ópticos. Aunque la agenesia del septum pellucidum fue la clave en este caso, no es un hallazgo presente en todos los pacientes.

Abstract Macrophages are pleiotropic and diverse cells that populate all tissues of the body. Besides tissue-specific resident macrophages such as alveolar macrophages, Kupffer cells, and microglia, multiple organs harbor at least two subtypes of other resident macrophages at steady state. During certain circumstances, like tissue insult, additional subtypes of macrophages are recruited to the tissue from the monocyte pool. Previously, a recruited macrophage population marked by expression of Spp1, Cd9, Gpnmb, Fabp5, and Trem2, has been described in several models of organ injury and cancer, and has been linked to fibrosis in mice and humans. Here, we show that Notch2 blockade, given systemically or locally, leads to an increase in this putative pro-fibrotic macrophage in the lung and that this macrophage state can only be adopted by monocytically derived cells and not resident alveolar macrophages. Using a bleomycin and COVID-19 model of lung injury and fibrosis, we find that the expansion of these macrophages before lung injury does not promote fibrosis but rather appears to ameliorate it. This suggests that these damage-associated macrophages are not, by themselves, drivers of fibrosis in the lung.

Abstract Microalgae are emerging as functional feed ingredients in aquaculture due to their immune-stimulating and stress-modulating properties. We investigated the potential of the microalgae Chlorella vulgaris as a feed supplement to improve the health and modulate microbiota and stress responses of Atlantic salmon. Triplicate groups of Atlantic salmon (~ 126 g) were reared in a recirculating aquaculture system (RAS) at 15 °C and received diets supplemented with 2% (CV2) or 14% (CV14) spray-dried C. vulgaris daily, 14% once weekly (CV14w), or a control diet (CD) for 8 weeks. Subsequently, all groups were exposed to an acute one-hour peracetic acid (CH3CO3H; PAA) treatment, a commonly used disinfectant in RAS. While CV14 increased feed conversion (FCR) significantly, feeding the diets CV2 and CV14w improved protein retention efficiency. CV14 significantly modulated beta-diversity in the intestinal digesta and mucosa, but this effect was already visible in fish fed CV2. Feeding CV14 and, to a lesser degree, CV2 increased the relative abundances of Paenarthrobacter and Trichococcus in the digesta and mucosa, which are able to metabolize complex carbohydrates. However, the same diets reduced the abundance of the lactic acid bacteria Lactobacillus and Weissella in the digesta and Floricoccus in the mucosa. Peracetic acid exposure induced systemic stress (increase in plasma glucose and cortisol) and a local immune response in the gill, with the most prominent upregulation of several immune- and stress-regulated genes (clra, cebpb, marco, tnfrsf14, ikba, c1ql2, drtp1) 18 h after exposure in fish fed the control diet. Fish receiving CV14 once a week showed a reduced transcriptional response to PAA exposure. Catalase protein abundance in the liver increased following exposure to PAA, while superoxide dismutase abundance in the gill and liver was increased in response to C. vulgaris inclusion before stress. Overall, the results highlight that a high (14%) inclusion rate of C. vulgaris in feed for Atlantic salmon impairs feed conversion and shifts the intestinal microbiota composition in digesta and mucosa. Weekly feeding of C. vulgaris proves a viable approach in improving protein retention and improving transcriptional resilience towards oxidative stress in increasingly intensive production systems. Thereby this study may motivate future studies on optimizing temporal feeding schedules for health-promoting aquafeeds.

Abstract Human pluripotent stem cells (hPSCs) represent an excellent cell source for regenerative medicine and tissue engineering applications. However, there remains a need for robust and scalable differentiation of stem cells into functional adult tissues. In this paper, we sought to address this challenge by developing magnetic microcapsules carrying hPSC spheroids. A co-axial flow-focusing microfluidic device was employed to encapsulate stem cells in core-shell microcapsules that also contained iron oxide magnetic nanoparticles (MNPs). These microcapsules exhibited excellent response to an external magnetic field and could be held at a specific location. As a demonstration of utility, magnetic microcapsules were used for differentiating hPSC spheroids as suspension cultures in a stirred bioreactor. Compared to standard suspension cultures, magnetic microcapsules allowed for more efficient media change and produced improved differentiation outcomes. In the future, magnetic microcapsules may enable better and more scalable differentiation of hPSCs into adult cell types and may offer benefits for cell transplantation.

Global climate change threatens ecosystem functioning worldwide. Forest ecosystems are particularly important for carbon sequestration, thereby buffering climate change and providing socio-economic services. However, recurrent stresses, such as heat waves, droughts and floods can affect forests with potential cascading effects on their carbon sink capacity, drought resilience and sustainability. Knowledge about the stress impact on the multitude of processes driving soil-plant-atmosphere interactions within these complex forest systems is widely lacking and uncertainty about future changes extremely high. Thus, forecasting forest response to climate change will require a dramatically improved process understanding of carbon and water cycling across various temporal (minutes to seasons) and spatial (leaf to ecosystem) scales covering atmosphere, biosphere, pedosphere and hydrosphere components.Many relevant processes controlling carbon and water exchange occur at small scales (e.g. rhizosphere, single leaf) with a high spatial and temporal variability, which is poorly constrained. However, interactions and feedback loops can be key players that amplify or dampen a system’s response to stress. Moreover, spatial and temporal scaling rules for these non-linear processes in structurally and functionally diverse ecosystems are unknown. Legacy effects, for example, altered response after previous stress and retarded recovery of forests after climate extremes, are not captured in state-of-the-art models. Currently, we are lacking the appropriate and interconnected measurement, data assimilation and modelling tools allowing for a comprehensive, real-time quantification of key processes at high spatio-temporal coverage in heterogeneous environments. Moreover, since climate impacts are highly unpredictable with respect to timing and location, future research will require novel mobile, easily deployable and cost-efficient approaches. ECOSENSE, therefore, assembles expertise from environmental and engineering sciences, both being excellently paired at the University of Freiburg.Our interdisciplinary research project will investigate all relevant scales in a next-generation ecosystem research assessment (ECOSENSE). Our vision is to detect and forecast critical changes in ecosystem functioning, based on the understanding of hierarchical process interaction. In the first phase, ECOSENSE will explore these process interactions by investigating pools and fluxes of water and carbon, i.e. CO2 exchange, isotope discrimination and volatile organic compounds (VOC), as well as stress indicators by remotely and in situ sensed chlorophyll fluorescence.To address these research tasks, ECOSENSE will develop, implement and test a distributed, autonomous, intelligent sensor network, based on novel microsensors tailored to the specific needs in remote and harsh forest environments. They will measure the spatio-temporal dynamics of ecosystem pools and fluxes in a naturally complex structured forest system with minimal physiological impact. Measured data will be transferred in real-time into a sophisticated database, which will be explored for process analysis, conducted by Artificial Intelligence and close to real-time process-based ecosystem models for now- and forecasting applications. Thereby, ECOSENSE will: i) break new ground for integrative ecosystem research by identifying hierarchies and interactions of abiotic and physiological processes of forest carbon and water exchange, ii) provide a profound understanding of complex ecosystem responses to environmental stressors and iii) enable the prediction of process-based alterations in ecosystem functioning and sustainability.Our novel ECOSENSE toolkit, tested and validated in controlled climate extreme experiments and our ECOSENSE Forest, will open new horizons for rapid assessment in vast and remote ecosystems. Thereby, ECOSENSE will allow for a unique avenue of data acquisition and, consequently, for unprecedented scale-crossing ecosystem understanding and modelling.

Abstract Ultrafast electron diffraction using MeV energy beams(MeV-UED) has enabled unprecedented scientific opportunities in the study of ultrafast structural dynamics in a variety of gas, liquid and solid state systems. Broad scientific applications usually pose different requirements for electron probe properties. Due to the complex, nonlinear and correlated nature of accelerator systems, electron beam property optimization is a time-taking process and often relies on extensive hand-tuning by experienced human operators. Algorithm based efficient online tuning strategies are highly desired. Here, we demonstrate multi-objective Bayesian active learning for speeding up online beam tuning at the SLAC MeV-UED facility. The multi-objective Bayesian optimization algorithm was used for efficiently searching the parameter space and mapping out the Pareto Fronts which give the trade-offs between key beam properties. Such scheme enables an unprecedented overview of the global behavior of the experimental system and takes a significantly smaller number of measurements compared with traditional methods such as a grid scan. This methodology can be applied in other experimental scenarios that require simultaneously optimizing multiple objectives by explorations in high dimensional, nonlinear and correlated systems.

We examined relations between foreign funding of U.S. colleges and universities and campus political developments. Seven studies investigated associations between foreign funding and campus liberal democratic norms, specifically, deterioration of free speech and academic freedom, and presence of antisemitism. Study I found that 349 colleges and universities received a total of almost $18 billion from foreign sources between 2014 and 2019. Study II examined relationships of foreign funding to campus deplatforming of speakers and punitive actions for speech protected by academic freedom. Main results were: 1. overall foreign funding was not strongly related to campus speech outcomes; 2. higher levels of deplatforming and speech punishment occurred on campuses that received funding from member states of the Organization of Islamic Cooperation and from authoritarian countries. Study III found weak evidence that foreign funding was associated with college students' reported exposure to antisemitic and anti-Zionist tropes. After demonstrating substantial correlations among three national measures of antisemitic incidents (Study IV), Study V found that foreign funding provided by member countries of the Organization for Islamic Cooperation or by authoritarian countries was associated with elevated levels of campus antisemitism and anti-Zionist incidents. Studies VI and VII found that antisemitic incidents on campus were associated with antisemitic incidents across the country. This research highlighted troubling possibilities about the potential role of foreign funding in higher education that deserve further investigation.

The stability of catalyst layers and membranes in proton exchange membrane water electrolysis (PEMWE) cells represents an ongoing challenge, compounded by the dissolution of components and migration of elements within the catalyst‐coated membrane (CCM). Conventional microscopy methods often struggle to efficiently evaluate large cross‐sections of PEMWE membranes, which is essential for representative analysis of technical scale CCMs. Herein, synchrotron radiation‐based X‐Ray fluorescence microscopy is exploited to analyze the stability of CCMs with around 1 μm resolution and a field of view of ≈200 × 75 μm2. Three application scenarios are investigated: 1) migration of catalyst elements, 2) dissolution of components, and 3) contaminated water supply containing Fe2+ ions. XRF is performed at three different X‐Ray energies (11.7, 11.4, and 11.0 keV), revealing the local elemental composition, including Pt, Ir, Ti, and Fe, under different stressing conditions. Notable observations include the distribution of Ir across the membrane and in the cathode catalyst layer, localization of Pt within the membrane, accumulation of Ti in the cathode catalyst layer, and minimal presence of Fe. XRF has been demonstrated to be a powerful analytical tool for accurate and high throughput imaging of catalyst degradation in PEMWE scenarios, particularly of technical scale devices.

IntroductionWorked examples support initial skill acquisition. They often show skill application on content knowledge from another, “exemplifying” domain (e.g., argumentation skills have to be applied to some contents). Although learners’ focus should remain on the skill, learners need to understand the content knowledge to benefit from worked examples. Previous studies relied on exemplifying domains that are familiar and contain simple topics, to keep learners’ focus on skill acquisition.AimWe examined whether using a relevant exemplifying domain would allow learners to acquire both skills and content knowledge simultaneously, or whether relevant content distracts from the main learning goal of skill acquisition.Methods and resultsIn a training study with 142 psychology students, we used example-based learning materials with an exemplifying domain that was either relevant or irrelevant for participants’ course outcomes. We assessed cognitive load, declarative knowledge about skills and course-related content knowledge, and argumentation quality. Incorporating relevant content knowledge in worked examples did not reduce learning outcomes compared to a condition using an irrelevant exemplifying domain.DiscussionContrary to previous research, the results suggest that worked examples with a relevant exemplifying domain could possibly be an efficient teaching method for fostering skills and content knowledge simultaneously.

Abstract Background Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a severely debilitating condition which markedly restricts activity and function of affected people. Since the beginning of the COVID-19 pandemic ME/CFS related to post-acute COVID-19 syndrome (PACS) can be diagnosed in a subset of patients presenting with persistent fatigue 6 months after a mostly mild SARS-CoV-2 infection by fulfillment of the Canadian Consensus Criteria (CCC 2003). Induction of autoimmunity after viral infection is a mechanism under intensive investigation. In patients with ME/CFS, autoantibodies against thyreoperoxidase (TPO), beta-adrenergic receptors (ß2AR), and muscarinic acetylcholine receptors (MAR) are frequently found, and there is evidence for effectiveness of immunomodulation with B cell depleting therapy, cyclophosphamide, or intravenous immunoglobulins (IVIG). Preliminary studies on the treatment of ME/CFS patients with immunoadsorption (IA), an apheresis that removes antibodies from plasma, suggest clinical improvement. However, evidence from placebo-controlled trials is currently missing. Methods In this double-blinded, randomized, sham-controlled, exploratory trial the therapeutic effect of five cycles of IA every other day in patients with ME/CFS, including patients with post-acute COVID-19 chronic fatigue syndrome (PACS-CFS), will be evaluated using the validated Chalder Fatigue Scale, a patient-reported outcome measurement. A total of 66 patients will be randomized at a 2:1 ratio: 44 patients will receive IA (active treatment group) and 22 patients will receive a sham apheresis (control group). Moreover, safety, tolerability, and the effect of IA on patient-reported outcome parameters, biomarker-related objectives, cognitive outcome measurements, and physical parameters will be assessed. Patients will be hospitalized at the clinical site from day 1 to day 10 to receive five IA treatments and medical visits. Four follow-up visits (including two visits at site and two visits via telephone call) at month 1 (day 30), 2 (day 60), 4 (day 120), and 6 (day 180; EOS, end of study visit) will take place. Discussion Although ME/CFS including PACS-CFS causes an immense individual, social, and economic burden, we lack efficient therapeutic options. The present study aims to investigate the efficacy of immunoadsorption and to contribute to the etiological understanding and establishment of diagnostic tools for ME/CFS. Trial registration Registration Number: NCT05710770 . Registered on 02 February 2023.

Abstract Background Lactic acid bacteria are commonly used as protective starter cultures in food products. Among their beneficial effects is the production of ribosomally synthesized peptides termed bacteriocins that kill or inhibit food-spoiling bacteria and pathogens, e.g., members of the Listeria species. As new bacteriocins and producer strains are being discovered rapidly, modern automated methods for strain evaluation and bioprocess development are required to accelerate screening and development processes. Results In this study, we developed an automated workflow for screening and bioprocess optimization for bacteriocin producing lactic acid bacteria, consisting of microcultivation, sample processing and automated antimicrobial activity assay. We implemented sample processing workflows to minimize bacteriocin adsorption to producer cells via addition of Tween 80 and divalent cations to the cultivation media as well as acidification of culture broth prior to cell separation. Moreover, we demonstrated the applicability of the automated workflow to analyze influence of media components such as MES buffer or yeast extract for bacteriocin producers Lactococcus lactis B1629 and Latilactobacillus sakei A1608. Conclusions Our automated workflow provides advanced possibilities to accelerate screening and bioprocess optimization for natural bacteriocin producers. Based on its modular concept, adaptations for other strains, bacteriocin products and applications are easily carried out and a unique tool to support bacteriocin research and bioprocess development is provided.

Summary: Background: Two important questions regarding the genetics of pancreatic adenocarcinoma (PDAC) are 1. Which germline genetic variants influence the incidence of this cancer; and 2. Whether PDAC causally predisposes to associated non-malignant phenotypes, such as type 2 diabetes (T2D) and venous thromboembolism (VTE). Methods: In this study of 8803 patients with PDAC and 67,523 controls, we first performed a large-scale transcriptome-wide association study to investigate the association between genetically determined gene expression in normal pancreas tissue and PDAC risk. Secondly, we used Mendelian Randomization (MR) to analyse the causal relationships among PDAC, T2D (74,124 cases and 824,006 controls) and VTE (30,234 cases and 172,122 controls). Findings: Sixteen genes showed an association with PDAC risk (FDR

Staphylococcus aureus is a common colonizer of the skin and nares of healthy individuals, but also a major cause of severe human infections. During interaction with the host, pathogenic bacteria must adapt to a variety of adverse conditions including nutrient deprivation. In particular, they encounter severe iron limitation in the mammalian host through iron sequestration by haptoglobin and iron-binding proteins, a phenomenon called “nutritional immunity.” In most bacteria, including S. aureus, the ferric uptake regulator (Fur) is the key regulator of iron homeostasis, which primarily acts as a transcriptional repressor of genes encoding iron acquisition systems. Moreover, Fur can control the expression of trans-acting small regulatory RNAs that play an important role in the cellular iron-sparing response involving major changes in cellular metabolism under iron-limiting conditions. In S. aureus, the sRNA IsrR is controlled by Fur, and most of its predicted targets are iron-containing proteins and other proteins related to iron metabolism and iron-dependent pathways. To characterize the IsrR targetome on a genome-wide scale, we combined proteomics-based identification of potential IsrR targets using S. aureus strains either lacking or constitutively expressing IsrR with an in silico target prediction approach, thereby suggesting 21 IsrR targets, of which 19 were negatively affected by IsrR based on the observed protein patterns. These included several Fe-S cluster- and heme-containing proteins, such as TCA cycle enzymes and catalase encoded by katA. IsrR affects multiple metabolic pathways connected to the TCA cycle as well as the oxidative stress response of S. aureus and links the iron limitation response to metabolic remodeling. In contrast to the majority of target mRNAs, the IsrR-katA mRNA interaction is predicted upstream of the ribosome binding site, and further experiments including mRNA half-life measurements demonstrated that IsrR, in addition to inhibiting translation initiation, can downregulate target protein levels by affecting mRNA stability.

Current therapy protocols fail to cure high-grade gliomas and prevent recurrence. Therefore, novel approaches need to be developed. A re-programing of glioma cell fate is an alternative attractive way to stop tumor growth. The two-step protocol applies the antiproliferative GQ bi-(AID-1-T) and small molecule inducers with BDNF to trigger neural differentiation into terminally differentiated cells, and it is very effective on GB cell cultures. This original approach is a successful example of the “differentiation therapy”. To demonstrate a versatility of this approach, in this publication we have extended a palette of cell cultures to gliomas of II, III and IV Grades, and proved an applicability of that version of differential therapy for a variety of tumor cells. We have justified a sequential mode of adding of GQIcombi components to the glioma cells. We have shown a significant retardation of tumor growth after a direct injection of GQIcombi into the tumor in rat brain, model 101/8. Thus, the proposed strategy of influencing on cancer cell growth is applicable to be further translated for therapy use.

Abstract Background Streptococcus pyogenes (group A streptococcus, GAS) causes a variety of diseases ranging from mild superficial infections of the throat and skin to severe invasive infections, such as necrotizing soft tissue infections (NSTIs). Tissue passage of GAS often results in mutations within the genes encoding for control of virulence (Cov)R/S two component system leading to a hyper-virulent phenotype. Dendritic cells (DCs) are innate immune sentinels specialized in antigen uptake and subsequent T cell priming. This study aimed to analyze cytokine release by DCs and other cells of monocytic origin in response to wild-type and natural covR/S mutant infections. Methods Human primary monocyte-derived (mo)DCs were used. DC maturation and release of pro-inflammatory cytokines in response to infections with wild-type and covR/S mutants were assessed via flow cytometry. Global proteome changes were assessed via mass spectrometry. As a proof-of-principle, cytokine release by human primary monocytes and macrophages was determined. Results In vitro infections of moDCs and other monocytic cells with natural GAS covR/S mutants resulted in reduced secretion of IL-8 and IL-18 as compared to wild-type infections. In contrast, moDC maturation remained unaffected. Inhibition of caspase-8 restored secretion of both molecules. Knock-out of streptolysin O in GAS strain with unaffected CovR/S even further elevated the IL-18 secretion by moDCs. Of 67 fully sequenced NSTI GAS isolates, 28 harbored mutations resulting in dysfunctional CovR/S. However, analyses of plasma IL-8 and IL-18 levels did not correlate with presence or absence of such mutations. Conclusions Our data demonstrate that strains, which harbor covR/S mutations, interfere with IL-18 and IL-8 responses in monocytic cells by utilizing the caspase-8 axis. Future experiments aim to identify the underlying mechanism and consequences for NSTI patients.

In recent times, both geopolitical challenges and the need to counteract climate change have led to an increase in generated renewable energy as well as an increased demand for clean electrical energy. The resulting variability of electricity production and demand as well as an overall demand increase, put additional stress on the existing grid infrastructure. This leads to strongly increased maintenance demands for distribution system operators (DSOs). Today, condition monitoring is used to address these challenges. Researchers have already explored solutions for monitoring critical assets like switchgear and circuit breakers. However, with a shrinking knowledgeable technical workforce and increasing maintenance requirements, mere monitoring is insufficient. Already today, DSOs ask for actionable recommendations, optimization strategies, and prioritization methods to manage the growing task backlog effectively. In this paper we propose a vision of a grid-level cognitive assistance system that translates the outcome of diagnosis and prognosis systems into actionable work tasks for the grid operator. The solution is highly interdisciplinary and based on empirical studies of real-world requirements. We also describe the related work relevant to the multi-disciplinary aspects and summarize the research gaps that need to be closed over the next years.

Abstract To understand neurological complications of COVID-19 better both acutely and for recovery, we measured markers of brain injury, inflammatory mediators, and autoantibodies in 203 hospitalised participants; 111 with acute sera (1–11 days post-admission) and 92 convalescent sera (56 with COVID-19-associated neurological diagnoses). Here we show that compared to 60 uninfected controls, tTau, GFAP, NfL, and UCH-L1 are increased with COVID-19 infection at acute timepoints and NfL and GFAP are significantly higher in participants with neurological complications. Inflammatory mediators (IL-6, IL-12p40, HGF, M-CSF, CCL2, and IL-1RA) are associated with both altered consciousness and markers of brain injury. Autoantibodies are more common in COVID-19 than controls and some (including against MYL7, UCH-L1, and GRIN3B) are more frequent with altered consciousness. Additionally, convalescent participants with neurological complications show elevated GFAP and NfL, unrelated to attenuated systemic inflammatory mediators and to autoantibody responses. Overall, neurological complications of COVID-19 are associated with evidence of neuroglial injury in both acute and late disease and these correlate with dysregulated innate and adaptive immune responses acutely.

Rail infrastructure plays an important role in fulfilling the demand for freight and passenger transportation. Increases in traffic volume, heavier axles and vehicles, higher speeds, and increasing climate extremes all contribute to the constant strain on the infrastructure. Due to their major importance in the transportation of people and freight, they are subject to continuous condition monitoring. This is an essential requirement for the selective planning of maintenance tasks and ultimately for safe and reliable operation. Various measuring systems have been developed for this purpose. These must measure precisely, quickly, and robustly under difficult conditions. Whether installed from mobile or stationary platforms, they have to cope with a wide range of ambient temperatures and lighting conditions, harsh environmental influences, and varying degrees of reflection. Despite these circumstances, railway operators require precise measurement data, high data densities even at high traveling speeds, and a user-friendly presentation of the results. Photogrammetry, laser scanning, and fiber optics are light-based measurement methods that are used in this sector. They are able to record with high precision rail infrastructure such as overhead contact systems, clearance profiles, rail tracks, and much more. This article provides an overview of the established and modern optical sensing methods, as well as the use of artificial intelligence as an evaluation method, and highlights their advantages and disadvantages.

The utilization of robust, pre-trained foundation models enables simple adaptation to specific ongoing tasks. In particular, the recently developed Segment Anything Model (SAM) has demonstrated impressive results in the context of semantic segmentation. Recognizing that data collection is generally time-consuming and costly, this research aims to determine whether the use of these foundation models can reduce the need for training data. To assess the models’ behavior under conditions of reduced training data, five test datasets for semantic segmentation will be utilized. This study will concentrate on traffic sign segmentation to analyze the results in comparison to Mask R-CNN: the field’s leading model. The findings indicate that SAM does not surpass the leading model for this specific task, regardless of the quantity of training data. Nevertheless, a knowledge-distilled student architecture derived from SAM exhibits no reduction in accuracy when trained on data that have been reduced by 95%.

As nutrient-rich water in aquaponic systems cannot supply growing plants with all the required trace elements, supplementation with specific fertilizers is performed to make up for this deficit. While chelated fertilizers such as ethylenediaminetetraacetic acid–zinc disodium complex (Zn-EDTA) are becoming more popular in this context for improving plant growth in aquaponic systems, little is known about their effects on fish. During two experiments, a total of 576 individuals of catfish fry (0.19 g) and fingerlings (220.01 g) of the African catfish (Clarias gariepinus; Burchell, 1822) were kept separately for 32 days under experimental aquarium conditions. The fry was exposed to 0.125 and 0.5 mg/L, while the fingerlings were exposed to 0.5 and 2.0 mg/L Zn-EDTA in a plantless aquaponic system. The third treatment group consisted of a control group without Zn-EDTA. The growth, mortality, and ethological indicators were assessed for all growth stages, while the leukocyte distribution and histopathological changes were additionally determined for the fingerlings. As the feed intake in the experiment was limited, the investigations were focused on the effects of Zn-EDTA and not on the growth process of a respective fish growth stage. While the growth, mortality, and behavior were not significantly different in both growing stages, the number of mature neutrophils changed significantly in all treatments in fingerlings. Zn was not detected in the histologically investigated organs at the tested concentrations using the staining method. However, morphological alterations of the gill epithelium were found on the secondary lamellae. Quantitative multiplex PCR was used to simultaneously evaluate the expression of 17 genes related to Zn metabolism and stress physiology in head kidney samples. The transcripts of several selected genes changed by up to 70-fold. Due to high individual variances, only the copy numbers of the KMT2A (lysine-specific methyltransferase 2a) gene were significantly different across treatment groups and sampling points. However, the present results indicate that the addition of Zn-EDTA at the tested concentrations can be considered relatively benign for the health and welfare of C. gariepinus, as no toxic effects of Zn-EDTA were observed in moderately hard to hard water.

BackgroundMobile health (mHealth) wearable devices are increasingly being adopted by individuals to help manage and monitor physiological signals. However, the current state of wearables does not consider the needs of racially minoritized low–socioeconomic status (SES) communities regarding usability, accessibility, and price. This is a critical issue that necessitates immediate attention and resolution. ObjectiveThis study’s aims were 3-fold, to (1) understand how members of minoritized low-SES communities perceive current mHealth wearable devices, (2) identify the barriers and facilitators toward adoption, and (3) articulate design requirements for future wearable devices to enable equitable access for these communities. MethodsWe performed semistructured interviews with low-SES Hispanic or Latine adults (N=19) from 2 metropolitan cities in the Midwest and West Coast of the United States. Participants were asked questions about how they perceive wearables, what are the current benefits and barriers toward use, and what features they would like to see in future wearable devices. Common themes were identified and analyzed through an exploratory qualitative approach. ResultsThrough qualitative analysis, we identified 4 main themes. Participants’ perceptions of wearable devices were strongly influenced by their COVID-19 experiences. Hence, the first theme was related to the impact of COVID-19 on the community, and how this resulted in a significant increase in interest in wearables. The second theme highlights the challenges faced in obtaining adequate health resources and how this further motivated participants’ interest in health wearables. The third theme focuses on a general distrust in health care infrastructure and systems and how these challenges are motivating a need for wearables. Lastly, participants emphasized the pressing need for community-driven design of wearable technologies. ConclusionsThe findings from this study reveal that participants from underserved communities are showing emerging interest in using health wearables due to the COVID-19 pandemic and health care access issues. Yet, the needs of these individuals have been excluded from the design and development of current devices.

Ultrasonic welding is an efficient method of joining thermoplastic fabrics or other textile semi-finished products in a watertight manner. It is applied in the making of functional clothing, such as chemical protective clothing, sportswear or smart clothing, or other technical products. Another special field of application developed at the Chair of Development and Assembly of Textile Products is the use of coated textiles as temperature control elements. For this type of product, it is necessary to design the ultrasonic welds in such a way that the media-tight coating is not damaged and the joint is continuous. This paper presents the option of evaluating and optimizing the ultrasonic welding process for the production of the textile tempering systems using transient thermal analysis in order to improve the overall quality of the seam and ensure media tightness along the seam. In the following article, the status of the welding process, in particular the ultrasonic process, the textile materials, the heating of the textile and the joining process of welding is presented. In addition, the transient heat flow through the textile is investigated with the aid of FEM methods; taking into account, various seam structures.

IntroductionCarp edema virus (CEV) is a fish poxvirus that primarily infects the gills of common carp. CEV causes koi sleepy disease (KSD), which is highly contagious and can result in mortality of up to 100%.MethodsIn the present study, we analyzed the stress and immune responses during KSD in two strains of common carp with different resistance to CEV: susceptible koi and resistant Amur sazan. Experiments were performed at two temperatures: 12°C and 18°C. In the case of koi carp, we also analyzed the effect of supplementation of 0.6% NaCl into tank water, which prevents mortality of the CEV-infected fish (salt rescue model).ResultsWe found that CEV-infected koi kept at 18°C had the highest viral load, which correlated with the most severe histopathological changes in the gills. CEV infection resulted in the activation of stress response reflected by the upregulated expression of genes involved in stress response in the stress axis organs and increased levels of cortisol and glucose in the blood plasma. These changes were the most pronounced in CEV-infected koi kept at 18°C. At both temperatures, the activation of antiviral immune response was observed in koi kept under freshwater and NaCl conditions upon CEV infection. Interestingly, a clear downregulation of the expression of adaptive immune genes was observed in CEV-infected koi kept under freshwater at 18°C.ConclusionCEV induces a stress response and modulates adaptive immune response in koi, and this is correlated with the level of viral load and disease development.

Objective: To further validate a serum proteomics panel for predicting radiographic (structural) knee OA progression. Design: Serum peptides were targeted by multiple-reaction-monitoring mass spectrometry in the New York University cohort (n ​= ​104). Knee OA progression was defined as joint space narrowing ≥1 in the tibiofemoral compartment of one knee per study participant over a 24-month follow-up. The discriminative ability of an 11-peptide panel was evaluated by multivariable logistic regression and area under the receiver operating characteristic curve (AUC), without and with demographic characteristics of age, sex, and body mass index. The association of each peptide with OA progression was assessed by odds ratios (OR) in multivariable logistic regression models adjusted for demographics. Results: The cohort included 46 (44%) knee OA progressors. The panel of 11 peptides alone yielded AUC ​= ​0.66 (95% CI [0.55, 0.77]) for discriminating progressors from non-progressors; demographic traits alone yielded AUC ​= ​0.66 (95% CI [0.55, 0.77]). Together the 11 peptides and demographics yielded AUC ​= ​0.72 (95% CI [0.62, 0.83]). CRAC1 had the highest odds for predicting OA progression (OR 2.014, 95% CI [0.996, 4.296], p ​= ​0.058). Conclusions: We evaluated a parsimonious serum proteomic panel and found it to be a good discriminator of knee radiographic OA progression from non-progression. Since these biomarkers are quantifiable in serum, they could be deployed relatively easily to provide a simple, cost-effective strategy for identifying and monitoring individuals at high risk of knee OA progression.

Abstract The intestinal lumen is filled with diverse chemical and physical stimuli. Intestinal epithelial cells sense these stimuli and signal to enteric neurons which coordinate a range of physiologic processes required for normal digestive tract function. Yet, the neuro-epithelial connections remain poorly resolved, in part because the tools for orchestrating interactions between these cellular compartments are lacking. We describe the development of a two-compartment microfluidic device for co-culturing enteric neurons with intestinal epithelial cells. The device contains epithelial and neuronal compartments connected by microgrooves. The epithelial compartment was designed for cell seeding via injection and confinement of intestinal epithelial cells derived from human intestinal organoids. We demonstrated that organoids planarized effectively and retained epithelial phenotype for over a week. In the second chamber we dissociated and cultured intestinal myenteric neurons including intrinsic primary afferent neurons (IPANs) from transgenic mice that expressed the fluorescent protein tdTomato. IPANs extended projections into microgrooves, surrounded and frequently made contacts with epithelial cells. The density and directionality of neuronal projections were enhanced by the presence of epithelial cells in the adjacent compartment. Our microfluidic device represents a platform that may, in the future, be used to dissect structure and function of neuro-epithelial connections in the gut and other organs (skin, lung, bladder, and others) in health and disease.

The ability to maintain functional hepatocytes has important implications for bioartificial liver development, cell-based therapies, drug screening, and tissue engineering. Several approaches can be used to restore hepatocyte function in vitro, including coating a culture substrate with extracellular matrix (ECM), encapsulating cells within biomimetic gels (Collagen- or Matrigel-based), or co-cultivation with other cells. This paper describes the use of bioactive heparin-based core-shell microcapsules to form and cultivate hepatocyte spheroids. These microcapsules are comprised of an aqueous core that facilitates hepatocyte aggregation into spheroids and a heparin hydrogel shell that binds and releases growth factors. We demonstrate that bioactive microcapsules retain and release endogenous signals thus enhancing the function of encapsulated hepatocytes. We also demonstrate that hepatic function may be further enhanced by loading exogenous hepatocyte growth factor (HGF) into microcapsules and inhibiting transforming growth factor (TGF)-β1 signaling. Overall, bioactive microcapsules described here represent a promising new strategy for the encapsulation and maintenance of primary hepatocytes and will be beneficial for liver tissue engineering, regenerative medicine, and drug testing applications.

Abstract The human polysialyltransferases ST8Sia II and ST8Sia IV catalyze the transfer of several Neu5Ac residues onto glycoproteins forming homopolymers with essential roles during different physiological processes. In salmonids, heterogeneous set of sialic acids polymers have been described in ovary and on eggs cell surface and three genes st8sia4, st8sia2-r1 and st8sia2-r2 were identified that could be implicated in these heteropolymers. The three polysialyltransferases from the salmonid Coregonus maraena were cloned, recombinantly expressed in HEK293 cells and the ST8Sia IV was biochemically characterized. The MicroPlate Sialyltransferase Assay and the non-natural donor substrate CMP-SiaNAl were used to demonstrate enzyme activity and optimize polysialylation reactions. Polysialylation was also carried out with natural donor substrates CMP-Neu5Ac, CMP-Neu5Gc and CMP-Kdn in cell-free and cell-based assays and structural analyses of polysialylated products using the anti-polySia monoclonal antibody 735 and endoneuraminidase N and HPLC approaches. Our data highlighted distinct specificities of human and salmonid polysialyltransferases with notable differences in donor substrates use and the capacity of fish enzymes to generate heteropolymers. This study further suggested an evolution of the biological functions of polySia. C. maraena ST8Sia IV of particular interest to modify glycoproteins with a variety of polySia chains.

Abstract Background About 50% of older patients with acute myeloid leukemia (AML) fail to attain complete remission (CR) following cytarabine plus anthracycline-based induction therapy. Salvage chemotherapy regimens are based on high-dose cytarabine (HiDAC), which is frequently combined with mitoxantrone (HAM regimen). However, CR rates remain low, with less than one-third of the patients achieving a CR. FLT3-ITD has consistently been identified as an unfavorable molecular marker in both relapsed and refractory (r/r)-AML. One-quarter of patients who received midostaurin are refractory to induction therapy and relapse rate at 2 years exceeds 40%. The oral second-generation bis-aryl urea tyrosine kinase inhibitor quizartinib is a very selective FLT3 inhibitor, has a high capacity for sustained FLT3 inhibition, and has an acceptable toxicity profile. Methods In this multicenter, upfront randomized phase II trial, all patients receive quizartinib combined with HAM (cytarabine 3g/m2 bidaily day one to day three, mitoxantrone 10mg/m2 days two and three) during salvage therapy. Efficacy is assessed by comparison to historical controls based on the matched threshold crossing approach with achievement of CR, complete remission with incomplete hematologic recovery (CRi), or complete remission with partial recovery of peripheral blood counts (CRh) as primary endpoint. During consolidation therapy (chemotherapy and allogeneic hematopoietic cell transplantation), patients receive either prophylactic quizartinib therapy or measurable residual disease (MRD)-triggered preemptive continuation therapy with quizartinib according to up-front randomization. The matched threshold crossing approach is a novel study-design to enhance the classic single-arm trial design by including matched historical controls from previous clinical studies. It overcomes common disadvantages of single-armed and small randomized studies, since the expected outcome of the observed study population can be adjusted based on the matched controls with a comparable distribution of known prognostic and predictive factors. Furthermore, balanced treatment groups lead to stable statistical models. However, one of the limitations of our study is the inability to adjust for unobserved or unknown confounders. Addressing the primary endpoint, CR/CRi/CRh after salvage therapy, the maximal sample size of 80 patients is assessed generating a desirable power of the used adaptive design, assuming a logistic regression is performed at a one-sided significance level α=0.05, the aspired power is 0.8, and the number of matching partners per intervention patient is at least 1. After enrolling 20 patients, the trial sample size will be recalculated in an interim analysis based on a conditional power argument. Conclusion Currently, there is no commonly accepted standard for salvage chemotherapy treatment. The objective of the salvage therapy is to reduce leukemic burden, achieve the best possible remission, and perform a hemopoietic stem-cell transplantation. Thus, in patients with FLT3-ITD mutation, the comparison of quizartinib with intensive salvage therapy versus chemotherapy alone appears as a logical consequence in terms of efficacy and safety. Ethics and dissemination Ethical approval and approvals from the local and federal competent authorities were granted. Trial results will be reported via peer-reviewed journals and presented at conferences and scientific meetings. Trial registration ClinicalTrials.gov NCT03989713; EudraCT Number: 2018-002675-17.

Image-based 3D reconstruction enables laparoscopic applications as image-guided navigation and (autonomous) robot-assisted interventions, which require a high accuracy. The review’s purpose is to present the accuracy of different techniques to label the most promising. A systematic literature search with PubMed and google scholar from 2015 to 2023 was applied by following the framework of “Review articles: purpose, process, and structure”. Articles were considered when presenting a quantitative evaluation (root mean squared error and mean absolute error) of the reconstruction error (Euclidean distance between real and reconstructed surface). The search provides 995 articles, which were reduced to 48 articles after applying exclusion criteria. From these, a reconstruction error data set could be generated for the techniques of stereo vision, Shape-from-Motion, Simultaneous Localization and Mapping, deep-learning, and structured light. The reconstruction error varies from below one millimeter to higher than ten millimeters—with deep-learning and Simultaneous Localization and Mapping delivering the best results under intraoperative conditions. The high variance emerges from different experimental conditions. In conclusion, submillimeter accuracy is challenging, but promising image-based 3D reconstruction techniques could be identified. For future research, we recommend computing the reconstruction error for comparison purposes and use ex/in vivo organs as reference objects for realistic experiments.

Cell–cell interactions between fibroblasts and immune cells, like macrophages, are influenced by interaction with the surrounding extracellular matrix during wound healing. In vitro hydrogel models that mimic and modulate these interactions, especially of soluble mediators like cytokines, may allow for a more detailed investigation of immunomodulatory processes. In the present study, a biomimetic extracellular matrix model based on fibrillar 3D collagen I networks with a functionalization with heparin or 6-ON-desulfated heparin, as mimics of naturally occurring heparan sulfate, was developed to modulate cytokine binding effects with the hydrogel matrix. The constitution and microstructure of the collagen I network were found to be stable throughout the 7-day culture period. A coculture study of primary human fibroblasts/myofibroblasts and M-CSF-stimulated macrophages was used to show its applicability to simulate processes of progressed wound healing. The quantification of secreted cytokines (IL-8, IL-10, IL-6, FGF-2) in the cell culture supernatant demonstrated the differential impact of glycosaminoglycan functionalization of the collagen I network. Most prominently, IL-6 and FGF-2 were shown to be regulated by the cell culture condition and network constitution, indicating changes in paracrine and autocrine cell–cell communication of the fibroblast–macrophage coculture. From this perspective, we consider our newly established in vitro hydrogel model suitable for mechanistic coculture analyses of primary human cells to unravel the role of extracellular matrix factors in key events of tissue regeneration and beyond.

The electric arc furnace (EAF) is considered the second most important process for the production of crude steel and is usually used for the melting of scrap. With the current emphasis on defossilization, its share in global steelmaking is likely to further increase. Due to the large production quantities, minor improvements to the EAF process can still accumulate into a significant reduction in overall energy and resource consumption. A major aspect in the efficient operation of the EAF is achieving beneficial slag properties, as the slag influences the composition of the steel and can reduce energy losses as well as the maintenance cost. In order to investigate the EAF operation, a dynamic process model is applied. Within the model, the chemical reactions of the metal–slag system are calculated based on the activities of the involved species. In this regard, multiple models for the calculation of the chemical activities have been implemented. However, depending on the chosen model, the computation of the slag activities can be computationally demanding. For this reason, the application of a neural network for the calculation of the chemical activities within the slag is investigated. The performance of the neural network is then compared to the results of the previously applied models by using the commercial software FactSage as a reference. The validation shows that the surrogate model achieves great accuracy while keeping the computation demand low.

Abstract Background Nutrition has not only an impact on the general wellbeing of an animal but can also affect reproductive processes. In cattle, feeding regimes can influence the age of puberty onset and alter gonadal development. We analyzed effects of different milk replacer (MR) feeding regimes during rearing on ovarian physiology with specific emphasis on the numbers as well as gene expression characteristics of granulosa cells (GCs) at the age of puberty onset. Two groups of calves received either 10% or 20% of bodyweight MR per day during their first 8 weeks. After weaning, both groups were fed the same mixed ration ad libitum until slaughter at 8 months. Results Animals of the 20% feeding group had a significantly higher body weight, but the proportion of animals having a corpus luteum at the time of slaughter was not different between groups, suggesting a similar onset of puberty. Calves of the 10% group showed a constant GC count regardless of the number of follicles (r = 0.23) whereas in the 20% group increasing numbers of GCs were detected with a higher follicle count (r = 0.71). As a first effort to find a possible molecular explanation for this unexpected limitation of GC numbers in the 10% group, we comparatively analyzed GC transcriptomes in both diet groups. The mRNA microarray analysis revealed a total of 557 differentially expressed genes comparing both groups (fold change > |1.5| and p

Abstract Ketamine is a rapid-acting antidepressant that also influences neural reactivity to affective stimuli. However, the effect of ketamine on behavioral affective reactivity is yet to be elucidated. The affect-modulated startle reflex paradigm (AMSR) allows examining the valence-specific aspects of behavioral affective reactivity. We hypothesized that ketamine alters the modulation of the startle reflex during processing of unpleasant and pleasant stimuli and weakens the resting-state functional connectivity (rsFC) within the modulatory pathway, namely between the centromedial nucleus of the amygdala and nucleus reticularis pontis caudalis. In a randomized, double-blind, placebo-controlled, cross-over study, thirty-two healthy male participants underwent ultra-high field resting-state functional magnetic resonance imaging at 7 T before and 24 h after placebo and S-ketamine infusions. Participants completed the AMSR task at baseline and one day after each infusion. In contrast to our hypothesis, ketamine infusion did not impact startle potentiation during processing of unpleasant stimuli but resulted in diminished startle attenuation during processing of pleasant stimuli. This diminishment significantly correlated with end-of-infusion plasma levels of ketamine and norketamine. Furthermore, ketamine induced a decrease in rsFC within the modulatory startle reflex pathway. The results of this first study on the effect of ketamine on the AMSR suggest that ketamine might attenuate the motivational significance of pleasant stimuli in healthy participants one day after infusion.

Microorganisms have the potential to address environmental pollution, but the interaction mechanism between microorganisms and mine tailings is not well understood. This work was aimed at determining the bacterial isolates in soils and mine tailings and evaluating the distribution of metals, antimony (Sb), and arsenic (As) in the soils around the Komsomolsk tailings. Areas with high concentrations of As, Sb, cadmium (Cd), and lead (Pb) were found. Assessment based on the value of the contamination factor (CF) indicated large-scale As, Sb, Pb, Cd, iron (Fe), bismuth (Bi), and beryllium (Be) pollution, especially in soils sampled from the northeast direction of the mine tailings. Soils had a higher number of CFUs per g of dry weight than did the tailings, ranging from 84 × 106 to 3.1 × 109 and from 20 × 106 to 1.7 × 109, respectively. Arsenic exhibited a positive statistical correlation with the number of CFUs of Agrococcus and Staphylococcus. In addition, a positive correlation was found between the concentration of Co and the number of CFUs of Moraxella and Microbacterium. The Sb exhibited a positive correlation with Streptomyces. These results can be used to develop methods for waste reclamation, including the use of isolated bacterial strains for arsenic removal by precipitation.

Abstract Background Osteoarthritis (OA) is a chronic debilitating disease impacting a significant percentage of the global population. While there are numerous surgical and non-invasive interventions that can postpone joint replacement, there are no current treatments which can reverse the joint damage occurring during the pathogenesis of the disease. While many groups are investigating the use of stem cell therapies in the treatment of OA, we still don’t have a clear understanding of the role of these cells in the body, including heterogeneity of tissue resident adult mesenchymal progenitor cells (MPCs). Methods In the current study, we examined MPCs from the synovium and individuals with or without a traumatic knee joint injury and explored the chondrogenic differentiation capacity of these MPCs in vitro and in vivo. Results We found that there is heterogeneity of MPCs with the adult synovium and distinct sub-populations of MPCs and the abundancy of these sub-populations change with joint injury. Furthermore, only some of these sub-populations have the ability to effect cartilage repair in vivo. Using an unbiased proteomics approach, we were able to identify cell surface markers that identify this pro-chondrogenic MPC population in normal and injured joints, specifically CD82LowCD59+ synovial MPCs have robust cartilage regenerative properties in vivo. Conclusions The results of this study clearly show that cells within the adult human joint can impact cartilage repair and that these sub-populations exist within joints that have undergone a traumatic joint injury. Therefore, these populations can be exploited for the treatment of cartilage injuries and OA in future clinical trials.

Smoltification was found to impact both immune and stress responses of farmed Atlantic salmon (Salmo salar), but little is known about how salinity change affects salmon months after completed smoltification. Here, we examined (1) the effect of salinity change from brackish water to seawater on the stress and immune responses in Atlantic salmon and (2) evaluated if functional diets enriched with microalgae can mitigate stress- and immune-related changes. Groups of Atlantic salmon were fed for 8 weeks with different microalgae-enriched diets in brackish water and were then transferred into seawater. Samples of the head kidney, gill, liver and plasma were taken before seawater transfer (SWT), 20 h after SWT, and 2 weeks after SWT for gene-expression analysis, plasma biochemistry and protein quantification. The salmon showed full osmoregulatory ability upon transfer to seawater reflected by high nkaα1b levels in the gill and tight plasma ion regulation. In the gill, one-third of 44 investigated genes were reduced at either 20 h or 2 weeks in seawater, including genes involved in cytokine signaling (il1b) and antiviral defense (isg15, rsad2, ifit5). In contrast, an acute response after 20 h in SW was apparent in the head kidney reflected by increased plasma stress indicators and induced expression of genes involved in acute-phase response (drtp1), antimicrobial defense (camp) and stress response (hspa5). However, after 2 weeks in seawater, the expression of antiviral genes (isg15, rsad2, znfx1) was reduced in the head kidney. Few genes (camp, clra, c1ql2) in the gill were downregulated by a diet with 8% inclusion of Athrospira platensis. The results of the present study indicate that salinity change months after smoltification evokes molecular stress- and immune responses in Atlantic salmon. However, microalgae-enriched functional diets seem to have only limited potential to mitigate the related changes.

The mechanisms underlying Parkinson’s disease (PD) are complex and not fully understood, and the box-and-arrow model among other current models present significant challenges. This paper explores the potential role of the allocentric brain and especially its grid cells in several PD motor symptoms, including bradykinesia, kinesia paradoxa, freezing of gait, the bottleneck phenomenon, and their dependency on cueing. It is argued that central hubs, like the locus coeruleus and the pedunculopontine nucleus, often narrowly interpreted in the context of PD, play an equally important role in governing the allocentric brain as the basal ganglia. Consequently, the motor and secondary motor (e.g., spatially related) symptoms of PD linked with dopamine depletion may be more closely tied to erroneous computation by grid cells than to the basal ganglia alone. Because grid cells and their associated central hubs introduce both spatial and temporal information to the brain influencing velocity perception they may cause bradykinesia or hyperkinesia as well. In summary, PD motor symptoms may primarily be an allocentric disturbance resulting from virtual faulty computation by grid cells revealed by dopamine depletion in PD.

Abstract Patient‐derived cancer organoids (PDOs) hold considerable promise for personalizing therapy selection and improving patient outcomes. However, it is challenging to generate PDOs in sufficient numbers to test therapies in standard culture platforms. This challenge is particularly acute for pancreatic ductal adenocarcinoma (PDAC) where most patients are diagnosed at an advanced stage with non‐resectable tumors and where patient tissue is in the form of needle biopsies. Here the development and characterization of microfluidic devices for testing therapies using a limited amount of tissue or PDOs available from PDAC biopsies is described. It is demonstrated that microfluidic PDOs are phenotypically and genotypically similar to the gold‐standard Matrigel organoids with the advantages of 1) spheroid uniformity, 2) minimal cell number requirement, and 3) not relying on Matrigel. The utility of microfluidic PDOs is proven by testing PDO responses to several chemotherapies, including an inhibitor of glycogen synthase kinase (GSKI). In addition, microfluidic organoid cultures are used to test effectiveness of immunotherapy comprised of NK cells in combination with a novel biologic. In summary, our microfluidic device offers considerable benefits for personalizing oncology based on cancer biopsies and may, in the future, be developed into a companion diagnostic for chemotherapy or immunotherapy treatments.

Bacteriocins are antimicrobial peptides applied in food preservation and are interesting candidates as alternatives to conventional antibiotics or as microbiome modulators. Recently, we established Corynebacterium glutamicum as a suitable production host for various bacteriocins including garvicin Q (GarQ). Here, we establish secretion of GarQ by C. glutamicum via the Sec translocon achieving GarQ titers of about 7 mg L–1 in initial fermentations. At neutral pH, the cationic peptide is efficiently adsorbed to the negatively charged envelope of producer bacteria limiting availability of the bacteriocin in culture supernatants. A combination of CaCl2 and Tween 80 efficiently reduces GarQ adsorption to C. glutamicum. Moreover, cultivation in minimal medium supplemented with CaCl2 and Tween 80 improves GarQ production by C. glutamicum to about 15 mg L–1 but Tween 80 resulted in reduced GarQ activity at later timepoints. Using a reporter strain and proteomic analyses, we identified HtrA, a protease associated with secretion stress, as another potential factor limiting GarQ production. Transferring production to HtrA-deficient C. glutamicum K9 improves GarQ titers to close to 40 mg L–1. Applying conditions of low aeration prevented loss in activity at later timepoints and improved GarQ titers to about 100 mg L–1. This is about 50-fold higher than previously shown with a C. glutamicum strain employing the native GarQ transporter GarCD for secretion and in the range of levels observed with the native producer Lactococcus petauri B1726. Additionally, we tested several synthetic variants of GarQ and were able to show that exchange of the methionine in position 5 to a phenylalanine (GarQM5F) results in markedly increased activity against Lactococcus lactis and Listeria monocytogenes. In summary, our findings shed light on several aspects of recombinant GarQ production that may also be of relevance for production with natural producers and other bacteriocins.

ObjectiveTo assess the cost-effectiveness and budget impact of olaparib as a maintenance therapy in platinum-responsive, metastatic pancreatic cancer patients harboring a germline BRCA1/2 mutation, using the Swiss context as a model.MethodsBased on data from the POLO trial, published literature and local cost data, we developed a partitioned survival model of olaparib maintenance including full costs for BRCA1/2 germline testing compared to FOLFIRI maintenance chemotherapy and watch-and-wait. We calculated the incremental cost-effectiveness ratio (ICER) for the base case and several scenario analyses and estimated 5-year budget impact.ResultsComparing olaparib with watch-and wait and maintenance chemotherapy resulted in incremental cost-effectiveness ratios of CHF 2,711,716 and CHF 2,217,083 per QALY gained, respectively. The 5-year costs for the olaparib strategy in Switzerland would be CHF 22.4 million, of which CHF 11.4 million would be accounted for by germline BRCA1/2 screening of the potentially eligible population. This would amount to a budget impact of CHF 15.4 million (USD 16.9 million) versus watch-and-wait.ConclusionsOlaparib is not a cost-effective maintenance treatment option. Companion diagnostics are an equally important cost driver as the drug itself.

ABSTRACTThe iron-regulated surface determinant protein B (IsdB) of Staphylococcus aureus is involved in the acquisition of iron from hemoglobin. Moreover, IsdB elicits an adaptive immune response in mice and humans. Here, we show that IsdB also has impact on innate immunity. IsdB induces the release of proinflammatory cytokines, including IL-6 and IL-1β, in innate immune cells of humans and mice. In silico analysis and thermophoresis show that IsdB directly binds to TLR4 with high affinity. TLR4 sensing was essential for the IsdB-mediated production of IL-6, IL-1β, and other cytokines as it was abolished by blocking of TLR4-MyD88-IRAK1/4-NF-κB signaling. The release of IL-1β additionally required activation of the NLRP3 inflammasome. In human monocytes infected with live S. aureus, IsdB was necessary for maximal IL-1β release. Our studies identify S. aureus IsdB as a novel pathogen-associated molecular pattern that triggers innate immune defense mechanisms.IMPORTANCEThe prevalence of multidrug-resistant Staphylococcus aureus is of global concern, and vaccines are urgently needed. The iron-regulated surface determinant protein B (IsdB) of S. aureus was investigated as a vaccine candidate because of its essential role in bacterial iron acquisition but failed in clinical trials despite strong immunogenicity. Here, we reveal an unexpected second function for IsdB in pathogen-host interaction: the bacterial fitness factor IsdB triggers a strong inflammatory response in innate immune cells via Toll-like receptor 4 and the inflammasome, thus acting as a novel pathogen-associated molecular pattern of S. aureus. Our discovery contributes to a better understanding of how S. aureus modulates the immune response, which is necessary for vaccine development against the sophisticated pathogen.

Abstract Background Over the past decade, DNA methylation (DNAm)-based deconvolution methods that leverage cell-specific DNAm markers of immune cell types have been developed to provide accurate estimates of the proportions of leukocytes in peripheral blood. Immune cell phenotyping using DNAm markers, termed immunomethylomics or methylation cytometry, offers a solution for determining the body’s immune cell landscape that does not require fresh blood and is scalable to large sample sizes. Despite significant advances in DNAm-based deconvolution, references at the population level are needed for clinical and research interpretation of these additional immune layers. Here we aim to provide some references for immune populations in a group of multi-ethnic post-menopausal American women. Results We applied DNAm-based deconvolution to a large sample of post-menopausal women enrolled in the Women’s Health Initiative (baseline, N = 58) or the ancillary Long Life Study (WHI-LLS, N = 1237) to determine the reference ranges of 58 immune parameters, including proportions and absolute counts for 19 leukocyte subsets and 20 derived cell ratios. Participants were 50–94 years old at the time of blood draw, and N = 898 (69.3%) self-identified as White. Using linear regression models, we observed significant associations between age at blood draw and absolute counts and proportions of naïve B, memory CD4+, naïve CD4+, naïve CD8+, memory CD8+ memory, neutrophils, and natural killer cells. We also assessed the same immune profiles in a subset of paired longitudinal samples collected 14–18 years apart across N = 52 participants. Our results demonstrate high inter-individual variability in rates of change of leukocyte subsets over this time. And, when conducting paired t tests to test the difference in counts and proportions between the baseline visit and LLS visit, there were significant changes in naïve B, memory CD4+, naïve CD4+, naïve CD8+, memory CD8+ cells and neutrophils, similar to the results seen when analyzing the association with age in the entire cohort. Conclusions Here, we show that derived cell counts largely reflect the immune profile associated with proportions and that these novel methods replicate the known immune profiles associated with age. Further, we demonstrate the value this methylation cytometry approach can add as a potential application in epidemiological studies.

Abstract This review deals with the selection of the electric propulsion system (EPS) for the internationally developed and designed, primary nuclear-electric space tug International Nuclear Power and Propulsion System (INPPS). INPPS is scheduled for interplanetary missions to Mars and Jupiter moon Europa missions by the end of decade 2020. Regarding specific technical and mission parameters preselected electric thruster (ET) types, developed by international companies and institutions, are analysed, evaluated and investigated for a possible application as propulsion system (PS), the so-called CET (Cluster of Electric Thrusters). It is analysed whether solely electric thrusters, combined in an adequate CET, enable the envisaged interplanetary missions—robotic and astronautic/crewed with the INPPS flagship. Thruster clusters with strategic consortium considerations are analysed as a feasible PS of the INPPS. The studied CET consists of the following: (a) only European ETs, (b) combination of German and European ETs, (c) Japanese and European ETs or at least (d) Japanese, European and US thrusters. The main results are (1) Robotic and crewed INPPS mission to Mars/Europa are realizable with EPS only (no chemical propulsion is needed), (2) that every CET, except (c) of only Japanese and part of European thrusters, is capable to perform the main part of envisaged INPPS flagship mission orbit to Mars, back to Earth and to Jupiter/Europa moon.

Altered expression and function of the extracellular matrix protein PRG4 have been associated with osteoarthritis. Here, the authors show that mast cell tryptase β cleaves PRG4, resulting in a reduction of lubrication and activation of inflammation in this context.

In-situ splitting of the liver before extended resection has gained broad attention. This two-step procedure requires several measures to make an effective and safe procedure. Although the procedure is performed in many institutions, there is no consensus on a uniform technique. The two steps can be divided into different parts and a standardized technique may render the procedure safer and the results will be easier to evaluate. In this paper, we describe a detailed approach to in-situ splitting that allows making both procedures safe, avoids liver necrosis, and is easily reproducible. In the first procedure the portal branches to segments I and IV to VIII are divided, the arterial branches and bile ducts to these segments are preserved and encircled and the parenchyma between segments II/III and IVa/b is divided. This avoids necrosis and bile leaks of segments I and IV and avoids urgent completion operations. In particular, the handling of vital structures close to the dissection line seems important to us. Complete splitting and securing the right and middle hepatic vein will make the second step of this procedure a minimal-risk procedure at a stage where the patient is still recovering from the more demanding first step.

The Fusarium fungi is found in cereals and feedstuffs and may produce mycotoxins, which are secondary metabolites, such as the T-2 toxin (T-2). In this work, we explored the hepatotoxicity of T-2 using microfluidic 3D hepatic cultures. The objectives were: (i) exploring the benefits of microfluidic 3D cultures compared to conventional 3D cultures available commercially (Aggrewell plates), (ii) establishing 3D co-cultures of hepatic cells (HepG2) and stellate cells (LX2) and assessing T-2 exposure in this model, (iii) characterizing the induction of metabolizing enzymes, and (iv) evaluating inflammatory markers upon T-2 exposure in microfluidic hepatic cultures. Our results demonstrated that, in comparison to commercial (large-volume) 3D cultures, spheroids formed faster and were more functional in microfluidic devices. The viability and hepatic function decreased with increasing T-2 concentrations in both monoculture and co-cultures. The RT-PCR analysis revealed that exposure to T-2 upregulates the expression of multiple Phase I and Phase II hepatic enzymes. In addition, several pro- and anti-inflammatory proteins were increased in co-cultures after exposure to T-2.

This work explores the possibility of automating the aerial survey of bridges to generate high-resolution images necessary for digital damage inspection. High-quality unmanned aerial vehicle (UAV) based 3D reconstruction of bridges is an important step towards autonomous infrastructure inspection. However, the calculation of optimal camera poses remains challenging due to the complex structure of bridges and is therefore often conducted manually. This process is time-consuming and can lead to quality losses. Research in this field to automate this process is yet sparse and often requires high informative models of the bridge as the base for calculations, which are not given widely. Therefore, this paper proposes an automated camera pose calculation method solely based on an easily accessible polygon mesh of the bridge. For safe operation, point cloud data of the environment are used for automated ground detection and obstacle avoidance including vegetation. First, an initial set of camera poses is generated based on a voxelized mesh created in respect to the quality requirements for 3D reconstruction using defined camera specification. Thereafter, camera poses not fulfilling safety distances are removed and specific camera poses are added to increase local coverage quality. Evaluations of three bridges show that for diverse bridge types, near-complete coverage was achieved. Due to the low computational effort of the voxel approach, the runtime was kept to a minimum, even for large bridges. The subsequent algorithm is able to find alternative camera poses even in areas where the optimal pose could not be placed due to obstacles.

Chimeric antigen receptor (CAR)-T cell therapy is a groundbreaking immunotherapy for cancer. However, the intricate and costly manufacturing process remains a hurdle. Improving the transduction rate is a potential avenue to cut down costs and boost therapeutic efficiency. Peptide nanofibrils (PNFs) serve as one such class of transduction enhancers. PNFs bind to negatively charged virions, facilitating their active engagement by cellular protrusions, which enhances virion attachment to cells, leading to increased cellular entry and gene transfer rates. While first-generation PNFs had issues with aggregate formation and potential immunogenicity, our study utilized in silico screening to identify short, endogenous, and non-immunogenic peptides capable of enhancing transduction. This led to the discovery of an 8-mer peptide, RM-8, which forms PNFs that effectively boost T cell transduction rates by various retroviral vectors. A subsequent structure-activity relationship (SAR) analysis refined RM-8, resulting in the D4 derivative. D4 peptide is stable and assembles into smaller PNFs, avoiding large aggregate formation, and demonstrates superior transduction rates in primary T and NK cells. In essence, D4 PNFs present an economical and straightforward nanotechnological tool, ideal for refining ex vivo gene transfer in CAR-T cell production and potentially other advanced therapeutic applications.