Search

Objective: Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is associated with an enhanced risk for liver and cardiovascular diseases and mortality. NAFLD can progress from simple hepatic steatosis to non-alcoholic steatohepatitis (NASH). However, the mechanisms predisposing to this progression remain undefined. Notably, hepatic mitochondrial dysfunction is a common finding in patients with NASH. Due to a lack of appropriate experimental animal models, it has not been evaluated whether this mitochondrial dysfunction plays a causative role for the development of NASH. Methods: To determine the effect of a well-defined mitochondrial dysfunction on liver physiology at baseline and during dietary challenge, C57BL/6J-mtFVB/N mice were employed. This conplastic inbred strain has been previously reported to exhibit decreased mitochondrial respiration likely linked to a non-synonymous gene variation (nt7778 G/T) of the mitochondrial ATP synthase protein 8 (mt-ATP8). Results: At baseline conditions, C57BL/6J-mtFVB/N mice displayed hepatic mitochondrial dysfunction characterized by decreased ATP production and increased formation of reactive oxygen species (ROS). Moreover, genes affecting lipid metabolism were differentially expressed, hepatic triglyceride and cholesterol levels were changed in these animals, and various acyl-carnitines were altered, pointing towards an impaired mitochondrial carnitine shuttle. However, over a period of twelve months, no spontaneous hepatic steatosis or inflammation was observed. On the other hand, upon dietary challenge with either a methionine and choline deficient diet or a western-style diet, C57BL/6J-mtFVB/N mice developed aggravated steatohepatitis as characterized by lipid accumulation, ballooning of hepatocytes and infiltration of immune cells. Conclusions: We observed distinct metabolic alterations in mice with a mitochondrial polymorphism associated hepatic mitochondrial dysfunction. However, a second hit, such as dietary stress, was required to cause hepatic steatosis and inflammation. This study suggests a causative role of hepatic mitochondrial dysfunction in the development of experimental NASH. Keywords: NAFLD, Steatohepatitis, Mitochondrial gene polymorphism, Mitochondrial dysfunction, Lipid metabolism, TNFα

Laser Metal Deposition (LMD) offers new perspectives for the fabrication of metal matrix nanocomposites (MMnCs). Current methods to produce MMnCs by LMD systematically involve the premixing of the nanopowders and the micropowders or require in-situ strategies, thereby restricting the possibilities to adjust the nature, content and location of the nano-reinforcement during printing. The objective of this study is to overcome such restrictions and propose a new process approach by direct injection of nanoparticles into a metallic matrix. Alumina (n-Al2O3) nanoparticles were introduced into a titanium matrix by using two different direct dry injection modes in order to locally increase the hardness. Energy dispersive X-ray spectroscopy (EDS) analyses validate the successful incorporation of the n-Al2O3 at chosen locations. Optical and high resolution transmission electron microscopic (HR-TEM) observations as well as X-ray diffraction (XRD) analyses indicate that n-Al2O3 powders are partly or totally dissolved into the Ti melted pool leading to the in-situ formation of a composite consisting of fine α2 lamellar microstructure within a Ti matrix and a solid solution with oxygen. Mechanical tests show a significant increase in hardness with the increase of injected n-Al2O3 amount. A maximum of 620 HV was measured that is almost 4 times higher than the pure LMD-printed Ti structure.

Enteroendocrine cells (EEC) produce neuropeptides, which are crucially involved in the maintenance of the intestinal barrier. Hence, EEC dysfunction is suggested to be involved in the complex pathophysiology of inflammatory bowel disease (IBD), which is characterized by decreased intestinal barrier function. However, the underlying mechanisms for EEC dysfunction are not clear and suitable models for a better understanding are lacking. Here, we demonstrate that Carboxypeptidase E (CPE) is specifically expressed in EEC of the murine colon and ileum and that its deficiency is associated with reduced intestinal levels of Neuropeptide Y (NPY) and Peptide YY (PYY), which are both produced by EEC. Moreover, cpe-/- mice exhibit an aggravated course of DSS-induced chronic colitis compared to wildtype littermates. In addition, we observed elevated mucosal IL-6 and KC transcript levels already at baseline conditions in cpe-/- mice. Moreover, supernatants obtained from isolated intestinal crypts of cpe-/- mice lead to increased IL-6 and KC expression in MODE-K cells in the presence of LPS. This effect was reversible by co-administration of recombinant NPY, suggesting a CPE mediated immunosuppressive effect in the intestines by influencing the processing of specific neuropeptides. In this context, the chemotaxis of bone marrow derived macrophages towards respective supernatants was enhanced. In conclusion, our data point to an anti-inflammatory role of CPE in the intestine by influencing local cytokine levels and thus regulating the migration of myeloid immune cells into the mucosa. These findings highlight the importance of EEC for intestinal homeostasis and propose EEC as potential therapeutic targets in IBD.

Abstract Background Network theory is largely applied in real-world systems to assess landscape connectivity using empirical or theoretical networks. Empirical networks are usually built from discontinuous individual movement trajectories without knowing the effect of relocation frequency on the assessment of landscape connectivity while theoretical networks generally rely on simple movement rules. We investigated the combined effects of relocation sampling frequency and landscape fragmentation on the assessment of landscape connectivity using simulated trajectories and empirical high-resolution (1 Hz) trajectories of Alpine ibex (Capra ibex). We also quantified the capacity of commonly used theoretical networks to accurately predict landscape connectivity from multiple movement processes. Methods We simulated forager trajectories from continuous correlated biased random walks in simulated landscapes with three levels of landscape fragmentation. High-resolution ibex trajectories were reconstructed using GPS-enabled multi-sensor biologging data and the dead-reckoning technique. For both simulated and empirical trajectories, we generated spatial networks from regularly resampled trajectories and assessed changes in their topology and information loss depending on the resampling frequency and landscape fragmentation. We finally built commonly used theoretical networks in the same landscapes and compared their predictions to actual connectivity. Results We demonstrated that an accurate assessment of landscape connectivity can be severely hampered (e.g., up to 66% of undetected visited patches and 29% of spurious links) when the relocation frequency is too coarse compared to the temporal dynamics of animal movement. However, the level of landscape fragmentation and underlying movement processes can both mitigate the effect of relocation sampling frequency. We also showed that network topologies emerging from different movement behaviours and a wide range of landscape fragmentation were complex, and that commonly used theoretical networks accurately predicted only 30–50% of landscape connectivity in such environments. Conclusions Very high-resolution trajectories were generally necessary to accurately identify complex network topologies and avoid the generation of spurious information on landscape connectivity. New technologies providing such high-resolution datasets over long periods should thus grow in the movement ecology sphere. In addition, commonly used theoretical models should be applied with caution to the study of landscape connectivity in real-world systems as they did not perform well as predictive tools.

Abstract Onoceroids are a rare family of triterpenes. One representative onoceroid is ambrein, which is the main component of ambergris used as a traditional medicine. We have previously identified the onoceroid synthase, BmeTC, in Bacillus megaterium and succeeded in creating ambrein synthase by introducing mutations into BmeTC. Owing to the structural similarity of ambrein to vitamin D, a molecule with diverse biological activities, we hypothesized that some of the activities of ambergris may be induced by the binding of ambrein to the vitamin D receptor (VDR). We demonstrated the VDR binding ability of ambrein. By comparing the structure–activity relationships of triterpenes with both the VDR affinity and osteoclastic differentiation-promoting activity, we observed that the activity of ambrein was not induced via the VDR. Therefore, some of the activities of ambergris, but not all, can be attributed to its VDR interaction. Additionally, six unnatural onoceroids were synthesized using the BmeTC reactions, and these compounds exhibited higher VDR affinity than that of ambrein. Enzymatic syntheses of onoceroid libraries will be valuable in creating a variety of bioactive compounds beyond ambergris.

The integration of deep learning-based tools into diagnostic workflows is increasingly prevalent due to their efficiency and reproducibility in various settings. We investigated the utility of automated nuclear morphometry for assessing nuclear pleomorphism (NP), a criterion of malignancy in the current grading system in canine pulmonary carcinoma (cPC), and its prognostic implications. We developed a deep learning-based algorithm for evaluating NP (variation in size, i.e., anisokaryosis and/or shape) using a segmentation model. Its performance was evaluated on 46 cPC cases with comprehensive follow-up data regarding its accuracy in nuclear segmentation and its prognostic ability. Its assessment of NP was compared to manual morphometry and established prognostic tests (pathologists’ NP estimates (n = 11), mitotic count, histological grading, and TNM-stage). The standard deviation (SD) of the nuclear area, indicative of anisokaryosis, exhibited good discriminatory ability for tumor-specific survival, with an area under the curve (AUC) of 0.80 and a hazard ratio (HR) of 3.38. The algorithm achieved values comparable to manual morphometry. In contrast, the pathologists’ estimates of anisokaryosis resulted in HR values ranging from 0.86 to 34.8, with slight inter-observer reproducibility (k = 0.204). Other conventional tests had no significant prognostic value in our study cohort. Fully automated morphometry promises a time-efficient and reproducible assessment of NP with a high prognostic value. Further refinement of the algorithm, particularly to address undersegmentation, and application to a larger study population are required.

To gain insights into phosphorus (P) dynamics in soils and the ability to predict soil responses to varying fertilizer inputs, mass balance models prove to be valuable tools. In this study, a new dynamic mass balance model, PBalD8, was used to describe the change in extracted P in the A horizon of soils subjected to diverse fertilizer treatments over a period of 50 to 60 years in five soil fertility experiments. The model employed a Freundlich equation to describe soil-solution partitioning of P and assumed that acid-lactate-extractable P represented a labile pool of P in instant equilibrium with soil solution P. Additionally, oxalate-extractable inorganic P was presumed to comprise the sum of the labile and stable pools of P, with mass flux to and from the latter described by Fick’s first law. The model was evaluated using results from extractions and P K-edge XANES spectroscopy. Notably, organic P, as revealed by P K-edge XANES, did not substantially contribute to long-term changes in soil P content and was therefore excluded from consideration. In general, the model offered reasonable fits to the extracted P concentrations. However, for the P-depleted treatments, a prerequisite was that the P removal through harvest was lower compared to measurements. Conversely, in three of the soils, the modelled fertilizer inputs needed to be reduced to 70 % to 85 % of the known additions. These discrepancies may be attributed to the involvement of deeper soil horizons, including deep crop uptake and mixing with lower soil layers, although other factors such as lateral dispersion and inaccuracies in estimating applied fertilizers cannot be discounted. These results underscore the necessity of gaining a more comprehensive understanding of how deeper soil horizons influence P mass balances in agricultural soils. In one of the soils, Fjärdingslöv, P K-edge XANES results demonstrated the formation of calcium phosphate over time in the highest fertilization treatment, consistent with the model. Additionally, in two soils, Kungsängen and the P-depleted Vreta Kloster soil, the model predicted a significant contribution from mineral weathering. However, the PBalD8 model also projected higher P leaching rates than those observed, suggesting that the model may not fully capture this P output term.

ObjectiveWe apply the superiorization methodology to the constrained intensity-modulated radiation therapy (IMRT) treatment planning problem. Superiorization combines a feasibility-seeking projection algorithm with objective function reduction: The underlying projection algorithm is perturbed with gradient descent steps to steer the algorithm towards a solution with a lower objective function value compared to one obtained solely through feasibility-seeking.ApproachWithin the open-source inverse planning toolkit matRad, we implement a prototypical algorithmic framework for superiorization using the well-established Agmon, Motzkin, and Schoenberg (AMS) feasibility-seeking projection algorithm and common nonlinear dose optimization objective functions. Based on this prototype, we apply superiorization to intensity-modulated radiation therapy treatment planning and compare it with (i) bare feasibility-seeking (i.e., without any objective function) and (ii) nonlinear constrained optimization using first-order derivatives. For these comparisons, we use the TG119 water phantom, the head-and-neck and the prostate patient of the CORT dataset.Main resultsBare feasibility-seeking with AMS confirms previous studies, showing it can find solutions that are nearly equivalent to those found by the established piece-wise least-squares optimization approach. The superiorization prototype solved the linearly constrained planning problem with similar dosimetric performance to that of a general-purpose nonlinear constrained optimizer while showing smooth convergence in both constraint proximity and objective function reduction.SignificanceSuperiorization is a useful alternative to constrained optimization in radiotherapy inverse treatment planning. Future extensions with other approaches to feasibility-seeking, e.g., with dose-volume constraints and more sophisticated perturbations, may unlock its full potential for high performant inverse treatment planning.

A method to isolate and sequence individual nuclei from human and mouse muscle biopsies provides further insight into the mechanisms of dystrophin loss and repair, in the context of Duchenne muscular dystrophy.

Introduction: In the Belgian part of the North Sea, sand for beach and foreshore nourishments is extracted from the Hinder Banks, about 40 km offshore. The extraction is not screened for shells and other very coarse material, as is the case for sand extraction for industrial use. Intermittent but intensive sand extraction took place from 2012 to 2019 on the Oosthinder, and since 2019 on the Noordhinder.Methods: To better understand sediment variability during human-induced disturbance of the seabed, both sandbanks have been monitored using multibeam bathymetry and backscatter, and Reineck box cores on board of the Research Vessel Belgica A962. Acoustic and sediment data were preferably jointly collected within a one-week period to substantiate the interrelationships.Result: Very well-sorted medium sands with low backscatter values (−28 to −33 dB) are detected near the top of the sandbank, while (moderately) well-sorted coarser sands and shell fragments with high backscatter values (−20 to −24 dB) are detected in the deeper parts of the sandbank slope. Although natural conditions such as sandwave movement and storms may mask early sediment changes, 7 years of intermittent and intensive sand extraction on the upper parts of a gentle sandbank slope caused a seabed deepening of at least 2 m, a backscatter decrease between 5 and 8 dB, a reduction in carbonate content by 1%–5% and an increase in organic matter content by 1%–2%. Two years after the cessation of the operations, the carbonate content increased and organic matter content decreased by 1%–2%.Discussion: Natural variability of sediments remains dependent on depth and geomorphology in both undisturbed, as well as recently and longer disturbed sandbank systems. However, a spreading of the more homogeneous, very well-sorted medium sands with limited carbonate content from the sandbank crest to the upper parts of the sandbank slope was observed, as sand extraction progressed. Subsequently, possible first signs of a slow return to the original sediment characteristics were detected.

Duchenne Muscular Dystrophy (DMD)’s complex multi-system pathophysiology, coupled with the cost-prohibitive logistics of multi-year drug screening and follow-up, has hampered the pursuit of new therapeutic approaches. Here we conducted a systematic historical and text mining-based pilot feasibility study to explore the potential of established or previously tested drugs as prospective DMD therapeutic agents. Our approach utilized a Swanson linking-inspired method to uncover meaningful yet largely hidden deep semantic connections between pharmacologically significant DMD targets and drugs developed for unrelated diseases. Specifically, we focused on molecular target-based MeSH terms and categories as high-yield bioinformatic proxies, effectively tagging relevant literature with categorical metadata. To identify promising leads, we comprehensively assembled published reports from 2011 and sampling from subsequent years. We then determined the earliest year when distinct MeSH terms or category labels of the relevant cellular target were referenced in conjunction with the drug, as well as when the pertinent target itself was first conclusively identified as holding therapeutic value for DMD. By comparing the earliest year when the drug was identifiable as a DMD treatment candidate with that of the first actual report confirming this, we computed an Index of Delayed Discovery (IDD), which serves as a metric of Swanson-linked latent knowledge. Using these findings, we identified data from previously unlinked articles subsetted via MeSH-derived Swanson linking or from target classes within the DrugBank repository. This enabled us to identify new but untested high-prospect small-molecule candidates that are of particular interest in repurposing for DMD and warrant further investigations.

Muscle damage and fibro-fatty replacement of skeletal muscles is a main pathologic feature of Duchenne muscular dystrophy (DMD) with more proximal muscles affected earlier and more distal affected later in the disease course, suggesting that different skeletal muscle groups possess distinctive characteristics that influence their susceptibility to disease. To explore transcriptomic factors driving differential gene expression and modulating DMD skeletal muscle severity, we characterized the transcriptome of vastus lateralis (VL), a more proximal and susceptible muscle, relative to tibialis anterior (TA), a more distal and protected muscle, in 15 healthy individuals using bulk RNA sequencing to identify gene expression differences that may mediate their relative susceptibility to damage with loss of dystrophin. Matching single nuclei RNA sequencing data was generated for 3 of the healthy individuals, to infer cell composition in the bulk RNA sequencing dataset and to improve mapping of differentially expressed genes to their cell source of expression. A total of 3,410 differentially expressed genes were identified and mapped to cell type using single nuclei RNA sequencing of muscle, including long non-coding RNAs and protein coding genes. There was an enrichment of genes involved in calcium release from the sarcoplasmic reticulum, particularly in the myofibers and these myofiber genes were higher in the VL. There was an enrichment of genes in “Collagen-Containing Extracellular Matrix” expressed by fibroblasts, endothelial, smooth muscle and pericytes, with most genes higher in the TA, as well as genes in “Regulation Of Apoptotic Process” expressed across all cell types. Previously reported genetic modifiers were also enriched within the differentially expressed genes. We also identify 6 genes with differential isoform usage between the VL and TA. Lastly, we integrate our findings with DMD RNA sequencing data from the TA, and identify “Collagen-Containing Extracellular Matrix” and “Negative Regulation Of Apoptotic Process” as differentially expressed between DMD compared to healthy. Collectively, these findings propose novel candidate mechanisms that may mediate differential muscle susceptibility in muscular dystrophies and provide new insight into potential therapeutic targets.

Acute scrotal pain (ASP) requiring surgical exploration is common in the pediatric population, but little has been reported on this subject with regard to the adult population. The aim of this study was to investigate the demographic and clinical characteristics and outcomes of scrotal explorations performed on adult patients. Patients over 21 years of age who underwent surgical exploration for ASP with suspected testicular torsion (TT) at 14 French hospitals between January 2005 and December 2019 were included in this study. The main outcome measures were demographic characteristics, pathology found during scrotal exploration, and perioperative outcomes. Logistic regression was used to perform univariate and multivariate analyses to identify predictors of TT. Data for 1329 men were analyzed. The median age was 30 (interquartile range [IQR]: 25–35; range: 21–89) years. Regarding the clinical examination, 867 (65.2%) patients presented with an elevation of the testicle, 613 (46.1%) patients with scrotal edema or erythema, and 211 (15.9%) patients with nausea or vomiting. Operative findings identified TT in only 684 (51.5%) patients, epididymo-orchitis in 112 (8.4%) patients, a tumor in 16 (1.2%) patients, and no causes in 475 (35.7%) patients. Orchiectomy for nonviable testes was required in 101 (7.6%) patients. In multivariate analysis, an elevation of the testicle, erythema/swelling, and the presence of nausea/vomiting were found to be associated with the occurrence of TT. Testicular torsion is not exclusive to children and adolescents, so must be considered in males of any age with acute scrotal findings. However, one-third of scrotal explorations in adults did not lead to a diagnosis.