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Lytic polysaccharide monooxygenases (LPMOs) are monocopper enzymes of industrial and biological importance. In particular, LPMOs play important roles in fungal lifestyle. No inhibitors of LPMOs have yet been reported. In this study, a diverse library of 100 plant extracts was screened for LPMO activity-modulating effects. By employing protein crystallography and LC-MS we successfully identified a natural LPMO inhibitor. Extract screening revealed a significant LPMO inhibition by methanolic extract of Cinnamomum cassia (cinnamon), which inhibited LsAA9A LPMO from Lentinus similis in a concentration-dependent manner. With a notable exception, other microbial LPMOs from families AA9 and AA10 were also inhibited by this cinnamon extract. The polyphenol cinnamtannin B1 was identified as the inhibitory component by crystallography. Cinnamtannin B1 was bound to the surface of LsAA9A at two distinct binding sites, one close to the active site and another at a pocket on the opposite side of the protein. Independent characterization of cinnamon extract by LC-MS and subsequent activity measurements confirmed that the compound inhibiting LsAA9A was cinnamtannin B1. The results of this study show that specific natural LPMO inhibitors of plant origin exist in nature, providing the opportunity for future exploitation of such compounds within various biotechnological contexts. Lytic polysaccharide monooxygenases (LPMOs) are monocopper enzymes of industrial and biological importance. In particular, LPMOs play important roles in fungal lifestyle. No inhibitors of LPMOs have yet been reported.In this study, a diverse library of 100 plant extracts was screened for LPMO activity-modulating effects. By employing protein crystallography and LC–MS, we successfully identified a natural LPMO inhibitor.Extract screening revealed a significant LPMO inhibition by methanolic extract of Cinnamomum cassia (cinnamon), which inhibited LsAA9A LPMO from Lentinus similis in a concentration-dependent manner. With a notable exception, other microbial LPMOs from families AA9 and AA10 were also inhibited by this cinnamon extract. The polyphenol cinnamtannin B1 was identified as the inhibitory component by crystallography. Cinnamtannin B1 was bound to the surface of LsAA9A at two distinct binding sites: one close to the active site and another at a pocket on the opposite side of the protein. Independent characterization of cinnamon extract by LC–MS and subsequent activity measurements confirmed that the compound inhibiting LsAA9A was cinnamtannin B1.The results of this study show that specific natural LPMO inhibitors of plant origin exist in nature, providing the opportunity for future exploitation of such compounds within various biotechnological contexts.

The histidine brace (His-brace) is a copper-binding motif that is associated with both oxidative enzymes and proteinaceous copper chaperones. Here, we used biochemical and structural methods to characterize mutants of a His-brace-containing copper chaperone from Pseudomonas fluorescens (PfCopC). A total of 15 amino acid variants in primary and second-sphere residues were produced and characterized in terms of their copper binding and redox properties. PfCopC has a very high affinity for Cu(II) and also binds Cu(I). A high reorganization barrier likely prevents redox cycling and, thus, catalysis. In contrast, mutations in the conserved second-sphere Glu27 enable slow oxidation of ascorbate. The crystal structure of the variant E27A confirmed copper binding at the His-brace. Unexpectedly, Asp83 at the equatorial position was shown to be indispensable for Cu(II) binding in the His-brace of PfCopC. A PfCopC mutant that was designed to mimic the His-brace from lytic polysaccharide monooxygenase-like family X325 did not bind Cu(II), but was still able to bind Cu(I). These results highlight the importance of the proteinaceous environment around the copper His-brace for reactivity and, thus, the difference between enzyme and chaperone.

Purpose of Review This review provides an overview of the most recent robotic ultrasound systems that have contemporary emerged over the past five years, highlighting their status and future directions. The systems are categorized based on their level of robot autonomy (LORA). Recent Findings Teleoperating systems show the highest level of technical maturity. Collaborative assisting and autonomous systems are still in the research phase, with a focus on ultrasound image processing and force adaptation strategies. However, missing key factors are clinical studies and appropriate safety strategies. Future research will likely focus on artificial intelligence and virtual/augmented reality to improve image understanding and ergonomics. Summary A review on robotic ultrasound systems is presented in which first technical specifications are outlined. Hereafter, the literature of the past five years is subdivided into teleoperation, collaborative assistance, or autonomous systems based on LORA. Finally, future trends for robotic ultrasound systems are reviewed with a focus on artificial intelligence and virtual/augmented reality.

Ultrasound (US) imaging, in contrast to other image guidance techniques, offers the distinct advantage of providing volumetric image data in real-time (4D) without using ionizing radiation. The goal of this study was to perform the first quantitative comparison of three different 4D US systems with fast matrix array probes and real-time data streaming regarding their target tracking accuracy and system latency. Sinusoidal motion of varying amplitudes and frequencies was used to simulate breathing motion with a robotic arm and a static US phantom. US volumes and robot positions were acquired online and stored for retrospective analysis. A template matching approach was used for target localization in the US data. Target motion measured in US was compared to the reference trajectory performed by the robot to determine localization accuracy and system latency. Using the robotic setup, all investigated 4D US systems could detect a moving target with sub-millimeter accuracy. However, especially high system latency increased tracking errors substantially and should be compensated with prediction algorithms for respiratory motion compensation.

Over 70 million European pregnant women are smokers during their child-bearing years. Consumption of tobacco-containing products during pregnancy is associated with several negative behavioral outcomes for the offspring, including a higher susceptibility for the development of attention-deficit/hyperactive disorder (ADHD). In efforts to minimize fetal exposure to tobacco smoke, many women around the world switch to nicotine replacement therapies (NRTs) during the gestational period; however, prenatal nicotine exposure (PNE) in any form has been associated with alterations in cognitive processes, including learning, memory, and attention. These processes are controlled by glutamatergic signaling of hippocampal pyramidal neurons within the CA1 region, suggesting actions of nicotine on glutamatergic transmission in this region if present prenatally. Accordingly, we aimed to investigate hippocampal glutamatergic function following PNE treatment in NMRI mice employing molecular, cellular electrophysiology, and pharmacological approaches, as well as to evaluate cognition in the rodent continuous performance task (rCPT), a recently developed mouse task allowing assessment of learning, attention, and impulsivity. PNE induced increases in the expression levels of mRNA coding for different glutamate receptors and subunits within the hippocampus. Functional alterations in AMPA and NMDA receptors on CA1 pyramidal neurons of PNE mice were suggestive of higher GluA2-lacking and lower GluN2A-containing receptors, respectively. Finally, PNE was associated with reduced learning, attention, and enhanced impulsivity in the rCPT. Alterations in glutamatergic functioning in CA1 neurons parallel changes seen in the spontaneously hypertensive rat ADHD model and likely contribute to the lower cognitive performance in the rCPT.

Thermal treatment of electrostatic complexes, above the denaturation temperature of the protein, is one possible method to induce particle formation and improve their stability towards changes in environmental conditions. The purpose of this study was to examine the effect of thermal treatment on mixtures of commercial, food-grade potato protein (PP) and anionic polysaccharides. Mixed solutions (1 wt%) of PP and gum arabic (GA) or carboxymethylcellulose (CMC) were acidified (pH 9.5 to 3.5), homogenized (200/50 bar) and pasteurized (90 °C, 10 min). The physicochemical properties of the acidified and heated samples were compared on particle size, charge, composition, turbidity and stability. It was found that the acidification in itself initiated formation of PP—GA and PP—CMC microparticles (MP). Upon heating, PP–GA MPs became smaller (d4,3 from 2.5 to 1.3 μm), uniformer, and the amount of PP incorporated was increased with 33%, causing more than a doubling of the turbidity. Oppositely, PP–CMC MPs did not change upon heating; the particles remained small (d4,3 0.8 μm) and polydisperse, consisting of 2.5:1 PP:CMC. It was hypothesized that the heat resistance of PP—CMC MPs was caused by stronger electrostatic forces between PP and CMC, while the heat-induced salt and pH resistance of PP—GA particles indicated the formation of other types of interactions preserving the MPs. Negative zeta potentials suggested PS was present at the exterior of the MPs. The obtained results demonstrated PP's ability to create micron-sized particles with polysaccharides, which could potentially replace dairy proteins in various applications.

Background Lytic polysaccharide monooxygenases (LPMOs) are important industrial enzymes known for their catalytic degradation of recalcitrant polymers such as cellulose or chitin. Their activity can be measured by lengthy HPLC methods, while high-throughput methods are less specific. A fast and specific LPMO assay would simplify screening for new or engineered LPMOs and accelerate biochemical characterization. Results A novel LPMO activity assay was developed based on the production of the dye phenolphthalein (PHP) from its reduced counterpart (rPHP). The colour response of rPHP oxidisation catalysed by the cellulose-specific LPMO from Thermoascus aurantiacus (TaAA9A), was found to increase tenfold by adding dehydroascorbate (DHA) as a co-substrate. The assay using a combination of rPHP and DHA was tested on 12 different metallo-enzymes, but only the LPMOs catalysed this reaction. The assay was optimized for characterization of TaAA9A and showed a sensitivity of 15 nM after 30 min incubation. It followed apparent Michaelis–Menten kinetics with kcat = 0.09 s−1 and KM = 244 µM, and the assay was used to confirm stoichiometric copper–enzyme binding and enzyme unfolding at a temperature of approximately 60 °C. DHA, glutathione and fructose were found to enhance LPMO oxidation of rPHP and in the optimized assay conditions these co-substrates also enabled cellulose degradation. Conclusions This novel and specific LPMO assay can be carried out in a convenient microtiter plate format ready for high-throughput screening and enzyme characterization. DHA was the best co-substrate tested for oxidation of rPHP and this preference appears to be LPMO-specific. The identified co-substrates DHA and fructose are not normally considered as LPMO co-substrates but here they are shown to facilitate both oxidation of rPHP and degradation of cellulose. This is a rare example of a finding from a high-throughput assay that directly translate into enzyme activity on an insoluble substrate. The rPHP-based assay thus expands our understanding of LPMO catalysed reactions and has the potential to characterize LPMO activity in industrial settings, where usual co-substrates such as ascorbate and oxygen are depleted.

International audience; BACKGROUND: Patients who have had a stroke with unknown time of onset have been previously excluded from thrombolysis. We aimed to establish whether intravenous alteplase is safe and effective in such patients when salvageable tissue has been identified with imaging biomarkers. METHODS: We did a systematic review and meta-analysis of individual patient data for trials published before Sept 21, 2020. Randomised trials of intravenous alteplase versus standard of care or placebo in adults with stroke with unknown time of onset with perfusion-diffusion MRI, perfusion CT, or MRI with diffusion weighted imaging-fluid attenuated inversion recovery (DWI-FLAIR) mismatch were eligible. The primary outcome was favourable functional outcome (score of 0-1 on the modified Rankin Scale [mRS]) at 90 days indicating no disability using an unconditional mixed-effect logistic-regression model fitted to estimate the treatment effect. Secondary outcomes were mRS shift towards a better functional outcome and independent outcome (mRS 0-2) at 90 days. Safety outcomes included death, severe disability or death (mRS score 4-6), and symptomatic intracranial haemorrhage. This study is registered with PROSPERO, CRD42020166903. FINDINGS: Of 249 identified abstracts, four trials met our eligibility criteria for inclusion: WAKE-UP, EXTEND, THAWS, and ECASS-4. The four trials provided individual patient data for 843 individuals, of whom 429 (51%) were assigned to alteplase and 414 (49%) to placebo or standard care. A favourable outcome occurred in 199 (47%) of 420 patients with alteplase and in 160 (39%) of 409 patients among controls (adjusted odds ratio [OR] 1·49 [95% CI 1·10-2·03]; p=0·011), with low heterogeneity across studies (I(2)=27%). Alteplase was associated with a significant shift towards better functional outcome (adjusted common OR 1·38 [95% CI 1·05-1·80]; p=0·019), and a higher odds of independent outcome (adjusted OR 1·50 [1·06-2·12]; p=0·022). In the alteplase group, 90 (21%) patients were severely disabled or died (mRS score 4-6), compared with 102 (25%) patients in the control group (adjusted OR 0·76 [0·52-1·11]; p=0·15). 27 (6%) patients died in the alteplase group and 14 (3%) patients died among controls (adjusted OR 2·06 [1·03-4·09]; p=0·040). The prevalence of symptomatic intracranial haemorrhage was higher in the alteplase group than among controls (11 [3%] vs two [\textless1%], adjusted OR 5·58 [1·22-25·50]; p=0·024). INTERPRETATION: In patients who have had a stroke with unknown time of onset with a DWI-FLAIR or perfusion mismatch, intravenous alteplase resulted in better functional outcome at 90 days than placebo or standard care. A net benefit was observed for all functional outcomes despite an increased risk of symptomatic intracranial haemorrhage. Although there were more deaths with alteplase than placebo, there were fewer cases of severe disability or death. FUNDING: None.

Pharmacological treatment modalities for non-alcoholic fatty liver disease (NAFLD) and steatohepatitis (NASH) are scarce, and discoveries are challenged by lack of predictive animal models adequately reflecting severe human disease stages and co-morbidities such as obesity and type 2 diabetes. To mimic human NAFLD/NASH etiology, many preclinical models rely on specific dietary components, though metabolism may differ considerably between species, potentially affecting outcomes and limiting comparability between studies. Consequently, understanding the physiological effects of dietary components is critical for high translational validity. This study investigated the effects of high fat, cholesterol, and carbohydrate sources on NASH development and metabolic outcomes in guinea pigs. Diet groups (n = 8/group) included: low-fat low-starch (LF-LSt), low-fat high-starch (LF-HSt), high-fat (HF) or HF with 4.2%, or 8.4% sugar water supplementation. The results showed that caloric compensation in HF animals supplied with sugar water led to reduced feed intake and a milder NASH phenotype compared to HF. The HF group displayed advanced NASH, weight gain and glucose intolerance compared to LF-LSt animals, but not LF-HSt, indicating an undesirable effect of starch in the control diet. Our findings support the HF guinea pig as a model of advanced NASH and highlights the importance in considering carbohydrate sources in preclinical studies of NAFLD.

Many bacteria secrete toxic protein complexes that modify and disrupt essential processes in the infected cell that can lead to cell death. To conduct their action, these toxins often need to cross the cell membrane and reach a specific substrate inside the cell. The investigation of these protein complexes is essential not only for understanding their biological functions but also for the rational design of targeted drug delivery vehicles that must navigate across the cell membrane to deliver their therapeutic payload. Despite the immense advances in experimental techniques, the investigations of the toxin entry mechanism have remained challenging. Computer simulations are robust complementary tools that allow for the exploration of biological processes in exceptional detail. In this review, we first highlight the strength of computational methods, with a special focus on all-atom molecular dynamics, coarse-grained, and mesoscopic models, for exploring different stages of the toxin protein entry mechanism. We then summarize recent developments that are significantly advancing our understanding, notably of the glycolipid–lectin (GL-Lect) endocytosis of bacterial Shiga and cholera toxins. The methods discussed here are also applicable to the design of membrane-penetrating nanoparticles and the study of the phenomenon of protein phase separation at the surface of the membrane. Finally, we discuss other likely routes for future development.

Background Barrett's esophagus (BE)—intestinal metaplasia in the esophagus—may progress to low-grade dysplasia (LGD), high-grade dysplasia (HGD), and ultimately, invasive esophageal adenocarcinoma (EAC). The course of BE in immunosuppressed lung transplant recipients is unknown. Methods This study retrospectively reviewed the records of patients who underwent lung transplant at a single center, Norton Thoracic Institute in Phoenix, Arizona, between January 1, 2010 and October 31, 2016. Pretransplant esophagram, esophagogastroduodenoscopy, 24-hour pH monitoring, high-resolution manometry, and gastric emptying studies were analyzed. Results Of the 466 patients who underwent lung transplant during the study period, 54 (11.59%) had BE on pretransplant esophagogastroduodenoscopy. Of these, 1 patient had HGD before lung transplant. The median age of patients was 64 years (interquartile range, 58.25 to 68.75 years); 66.7% were men. Median follow-up duration was 29.48 months (interquartile range, 19.69 to 37.98 months). Sixteen of 54 patients (29.62%) underwent antireflux surgery after lung transplant. LGD or EAC developed in 3 patients during posttransplant surveillance. One patient had a diagnosis of HGD 24 months after retransplant. She underwent complete endoscopic ablation and was dysplasia-free for 5 months, but ultimately the condition recurred, and she underwent esophagectomy for invasive cancer. Two patients had a diagnosis of LGD 7 and 13 months after lung transplant and were successfully treated with radiofrequency ablation. The rate of progression to dysplasia or EAC was 2.3% per patient-year. Conclusions BE seems to be more prevalent in lung transplant recipients than in the general population. The study findings suggest that patients with BE have a higher risk of BE-to-EAC progression after lung transplant and that HGD may progress rapidly in immunosuppressed patients. More intensive surveillance endoscopy may be required in patients with BE after lung transplant.

Background: MicroRNAs (miRNAs) represent important post-transcriptional regulators with a dynamic expression profile during health and disease. Objectives: We explored the miRNA profile of human mast cells (MCs) during sen­sitization with IgE, during activation through IgE, and relat ed it to prostaglandin D2 synthesis and histamine release. Method: We investigated the expression pattern of 762 miRNAs during the IgE-mediated sensitization and activation of MCs cultured from CD133+ stem cells that were isolated from allergic asthmatic patients and nonatopic controls. Results: IgE-mediated sensitization increased the expression of miRNA-210 eight-fold. This increase was sustained during IgE-mediated MC activation. Furthermore, we confirmed the increase of the miRNA-132/212 cluster after MC activation. Predicted target genes of miRNA-210/132/212 were enriched in several pathways known to be involved in MC activation. Histamine release was significantly higher in MCs from allergic patients when compared to controls, and a number of miRNAs correlated with histamine release and prostaglandin D2 synthesis during MC activation. Conclusion: The miRNAs and analysis presented here can help to elucidate the role of miRNAs in mediator release during MC activation. We speculate that miRNA-210 could be important in MC sensitization that leads to allergic symptoms.

Non‐alcoholic fatty liver disease is becoming a major health burden, as prevalence increases and there are no approved treatment options. Thiazolidinediones target the nuclear receptor peroxisome proliferator‐activated receptor γ (PPARγ) and have been investigated in several clinical trials for their potential in treating non‐alcoholic fatty liver disease (NAFLD) and non‐alcoholic steatohepatitis (NASH). PPARγ has specialized roles in distinct tissues and cell types, and although the primary function of PPARγ is in adipose tissue, where the highest expression levels are observed, hepatic expression levels of PPARγ are significantly increased in patients with NAFLD. Thus, NAFLD patients receiving treatment with PPARγ agonists might have a liver response apart from the one in adipose tissue. Owing to the different roles of PPARγ, new treatment strategies include development of compounds harnessing the beneficial effects of PPARγ while restricting PPARγ unwanted effects such as adipogenesis resulting in weight gain. Furthermore, dual or pan agonists targeting two or more of the PPARs have shown promising results in pre‐clinical research and some are currently proceeding to clinical trials. This MiniReview explores adipose‐ and liver‐specific actions of PPARγ, and how this knowledge may contribute in the search for new treatment modalities in NAFLD/NASH.

Hepatic fibrosis is the primary predictor of mortality in patients with non-alcoholic steatohepatitis (NASH). In this process, the activated hepatic stellate cells (HSCs) constitute the principal cells responsible for the deposition of a fibrous extracellular matrix, thereby driving the hepatic scarring. HSC activation, migration, and proliferation are controlled by a complex signaling network involving growth factors, lipotoxicity, inflammation, and cellular stress. Conversely, the clearance of activated HSCs is a prerequisite for the resolution of the extracellular fibrosis. Hence, pathways regulating the fate of the HSCs may represent attractive therapeutic targets for the treatment and prevention of NASH-associated hepatic fibrosis. However, the development of anti-fibrotic drugs for NASH patients has not yet resulted in clinically approved therapeutics, underscoring the complex biology and challenges involved when targeting the intricate cellular signaling mechanisms. This narrative review investigated the mechanisms of activation and inactivation of HSCs with a focus on NASH-associated hepatic fibrosis. Presenting an updated overview, this review highlights key cellular pathways with potential value for the development of future treatment modalities.

OBJECTIVES: To evaluate diagnostic accuracy of 18F-fluorodeoxyglucose positron emission tomography/computed tomography (FDG PET/CT) compared to contrast-enhanced CT in assessment of inguinal lymph node (ILN) metastases, distant metastases and synchronous cancers in patients with penile squamous cell carcinoma (pSCC).PATIENTS AND METHODS: During a 4-year period, patients with pSCC were scheduled for FDG PET/CT prior to surgical treatment at two referral centres that manage all penile cancers in Denmark. The primary endpoint was diagnostic accuracy of FDG PET/CT and of CT alone with histopathology or Response Evaluation Criteria In Solid Tumors (RECIST) as reference.RESULTS: We evaluated 171 patients for distant metastases and synchronous incident cancers and examined 286 groins in 143 patients for LN metastases by FDG PET/CT. Six groins disclosed false negatives. FDG PET/CT sensitivity was 85.4% per patient. In 135 patients (270 groins), CT images were evaluated separately and 22 groins disclosed false negatives. CT sensitivity was 47.5% per patient. FDG PET/CT detected pSCC distant metastases in seven patients. Distant metastases from other cancers were newly detected in three patients. In eight patients, an incidental synchronous cancer was detected. Seven out of the 18 distant malignancies detected depended on FDG PET information.CONCLUSION: This study underlines the increased diagnostic accuracy of FDG PET/CT compared to CT alone in the evaluation of ILN status. In patients with palpable LNs, the advantage of FDG PET/CT over CT is less pronounced. FDG PET/CT may play a role in penile cancer evaluation.

Lytic polysaccharide monooxygenases (LPMOs) are mononuclear copper enzymes that catalyse the oxidative cleavage of glycosidic bonds. They are characterised by two histidine residues that coordinate copper in a configuration termed the Cu-histidine brace. Although first identified in bacteria and fungi, LPMOs have since been found in all biological kingdoms. LPMOs are now included in commercial enzyme cocktails used in industrial biorefineries. This has led to increased process yield due to the synergistic action of LPMOs with glycoside hydrolases. However, the introduction of LPMOs makes control of the enzymatic step in industrial stirred-tank reactors more challenging, and the operational stability of the enzymes is reduced. It is clear that much is still to be learned about the interaction between LPMOs and their complex natural and industrial environments, and fundamental scientific studies are required towards this end. Several atomic-resolution structures have been solved providing detailed information on the Cu-coordination sphere and the interaction with the polysaccharide substrate. However, the molecular mechanisms of LPMOs are still the subject of intense investigation; the key question being how the proteinaceous environment controls the copper cofactor towards the activation of the O-O bond in O2 and cleavage of the glycosidic bonds in polysaccharides. The need for biochemical characterisation of each putative LPMO is discussed based on recent reports showing that not all proteins with a Cu-histidine brace are enzymes.

Environmentally friendly sources of energy and chemicals are essential constituents of a sustainable society. An important step towards this goal is the utilization of non-edible biomass as supply of building blocks for future biorefineries. Lytic polysaccharide monooxygenases (LPMOs) are enzymes that play a critical role in breaking the chemical bonds in the most abundant polymers found in recalcitrant biomass, such as cellulose and chitin. Predicting optimal strategies for producing LPMOs is often non-trivial, and methods allowing for screening several strategies simultaneously are therefore needed. Here, we present a standardized platform for cloning LPMOs. The platform allows users to combine gene fragments with different expression vectors in a simple 15-minute reaction, thus enabling rapid exploration of several gene contexts, hosts and expression strategies in parallel. The open-source LyGo platform is accompanied by easy-to-follow online protocols for both cloning and expression. As a demonstration, we utilize the LyGo platform to explore different strategies for expressing several different LPMOs in Escherichia coli, Bacillus subtilis, and Komagataella phaffii.

Tools for automatic image analysis are gaining importance in the clinical workflow, ranging from time-saving tools in diagnostics to real-time methods in image-guided interventions. Over the last years, ultrasound (US) imaging has become a promising modality for image guidance due to its ability to provide volumetric images of soft tissue in real-time without using ionizing radiation. One key challenge in automatic US image analysis is the identification of suitable features to describe the image or regions within, e.g. for recognition, alignment or tracking tasks. In recent years, features that were learned data-drivenly provided promising results. Even though these approaches outperformed hand-crafted feature extractors in many applications, there is still a lack of feature learning for local description of three-dimensional US (3DUS) images. In this work, we present a completely data-driven feature learning approach for 3DUS images for usage in target tracking. To this end, we use a 3D convolutional autoencoder (AE) with a custom loss function to encode 3DUS image patches into a compact latent space that serves as a general feature description. For evaluation, we trained and tested the proposed architecture on 3DUS images of the liver and prostate of five different subjects and assessed the similarity between the decoded patches and the original ones. Subject-and organ-specific as well as general AEs are trained and evaluated. Specific AEs could reconstruct patches with a mean Normalized Cross Correlation of 0.85 and 0.81 at maximum in liver and prostate, respectively. It can also be shown that the AEs are transferable across subjects and organs, with a small accuracy decrease to 0.83 and 0.81 (liver, prostate) for general AEs. In addition, a first tracking study was performed to show feasibility of tracking in latent space. In this work, we could show that it is possible to train an AE that is transferable across two target regions and several subjects. Hence, convolutional AEs present a promising approach for creating a general feature extractor for 3DUS.

Lytic polysaccharide monooxygenase (LPMO) and copper binding protein CopC share a similar mononuclear copper site. This site is defined by an N-terminal histidine and a second internal histidine side chain in a configuration called the histidine brace. To understand better the determinants of reactivity, the biochemical and structural properties of a well-described cellulose-specific LPMO from Thermoascus aurantiacus (TaAA9A) is compared with that of CopC from Pseudomonas fluorescens (PfCopC) and with the LPMO-like protein Bim1 from Cryptococcus neoformans. PfCopC is not reduced by ascorbate but is a very strong Cu(II) chelator due to residues that interacts with the N-terminus. This first biochemical characterization of Bim1 shows that it is not redox active, but very sensitive to H2O2, which accelerates the release of Cu ions from the protein. TaAA9A oxidizes ascorbate at a rate similar to free copper but through a mechanism that produce fewer reactive oxygen species. These three biologically relevant examples emphasize the diversity in how the proteinaceous environment control reactivity of Cu with O2.

Phase separation of immiscible fluids is a common phenomenon in polymer chemistry, and is recognized as an important mechanism by which cells compartmentalize their biochemical reactions. Biomolecular condensates are condensed fluid droplets in cells that form by liquid-liquid phase separation of intrinsically-disordered proteins. They have a wide range of functions and are associated with chronic neurodegenerative diseases in which they become pathologically rigid. However, it remains unclear how their material properties depend on the molecular structure of the proteins. Here we explore the phase behaviour and structure of a model biomolecular condensate composed of semi-flexible polymers with attractive end-caps using coarse-grained simulations. The model contains the minimal molecular features that are sufficient to observe liquid-liquid phase separation of soluble polymers into a porous, three-dimensional network in which their end-caps reversibly bind at junctions. The distance between connected junctions scales with the polymer length as a self-avoiding random walk over a wide range of concentration with a weak affinity-dependent prefactor. By contrast, the average number of polymers that meet at the junctions depends on the end-cap affinity but only weakly on the polymer length. The structured porosity of the condensed phase suggests a mechanism for cells to regulate biomolecular condensates. Protein interaction sites may be turned on or off to modulate the condensate's porosity and therefore the diffusion and interaction of additional proteins. This journal is

Nonalcoholic steatohepatitis (NASH) is one of the most common chronic liver diseases in the world, yet no pharmacotherapies are available. The lack of translational animal models is a major barrier impeding elucidation of disease mechanisms and drug development. Multiple preclinical models of NASH have been proposed and can broadly be characterized as diet-induced, deficiency-induced, toxin-induced, genetically induced, or a combination of these. However, very few models develop advanced fibrosis while still reflecting human disease etiology or pathology, which is problematic since fibrosis stage is considered the best prognostic marker in patients and an important endpoint in clinical trials of NASH. While mice and rats predominate the NASH research, several other species have emerged as promising models. This review critically evaluates animal models of NASH, focusing on their ability to develop advanced fibrosis while maintaining their relevance to the human condition.

Variability in disease development due to differences in strains and breeders constitutes a substantial challenge in preclinical research. However, the impact of the breeder on non-alcoholic steatohepatitis (NASH) is not yet fully elucidated. This retrospective study investigates NASH development in guinea pigs from Charles River or Envigo fed a high fat diet (20% fat, 15% sucrose, 0.35% cholesterol) for 16 or 24/25 weeks. Charles River animals displayed more severe NASH, with higher steatosis (p Ccl2, Cxcl8) and fibrotic (Pdgf, Serpine1, Col1a1) genes was also increased (p

Introduction In animal models of non-alcoholic fatty liver disease (NAFLD), assessment of disease severity and treatment effects of drugs rely on histopathological scoring of liver biopsies. However, little is known about the sampling variation in liver samples from animal models of NAFLD, even though several histopathological hallmarks of the disease are known to be affected by sampling variation in patients. The aim of this study was to assess the sampling variation in multiple paired liver biopsies from three commonly used diet-induced rodent models of NAFLD. Methods Eight male C57BL/6 mice, 8 male Sprague Dawley rats and 16 female Hartley guinea pigs were fed a NAFLD-inducing high-fat diet for 16 weeks (mice and rats), 20 or 24 weeks (guinea pigs). After the initial diet period, liver sections were sampled and subsequently assessed by histopathological scoring and biochemical analyses. Results Fibrosis was heterogeneously distributed throughout the liver in mice, manifesting as both intra- and interlobular statistically significant differences. Hepatic triglyceride content showed interlobular differences in mice, and both intra- and interlobular differences in guinea pigs (24-week time point) all of which were statistically significant. Also, hepatic cholesterol content was subject to significant intra-lobular sampling variation in mice, and hepatic glycogen content differed significantly between lobes in mice and guinea pigs. Discussion Dependent on animal model, both histopathological and biochemical end-points differed between sampling sites in the liver. Based on these findings, we recommend that sample site location is highly standardized and properly reported in order to minimize potential sampling variation and to optimize reproducibility and meaningful comparisons of preclinical studies of NAFLD.

Background This randomized, double‐blind trial was conducted to determine the optimal dose for clinical efficacy of the SQ tree SLIT‐tablet. An environmental exposure chamber (EEC) was used to reduce variability of allergen exposure and allow investigation of symptom reduction towards different species from the birch homologous group in separate EEC sessions. Methods Eligible subjects (N = 219) were randomized to receive treatment with placebo or the SQ tree SLIT‐tablet (2, 7, or 12 DU) for 24 weeks. EEC pollen challenges were conducted outside the birch pollen season and included four birch and two oak EEC sessions. The primary efficacy endpoint was the average allergic rhinoconjunctivitis (ARC) total symptom score (TSS) after 24 weeks of treatment. Results There was a statistically significantly lower TSS during the 24‐week birch EEC session for 7 DU and 12 DU compared to placebo with relative differences of 24% (P = 0.03) and 25% (P = 0.02). For the 24‐week oak EEC session, there was a statistically significant difference for 12 DU (24%, P = 0.03). IgE and IgG4 measurements supported these findings and demonstrated cross‐reactivity to all other species within the birch homologous group. Treatment was well‐tolerated with the most frequently reported adverse reactions being the local reactions in the oral cavity of mild‐to‐moderate severity. Conclusion This trial demonstrates that the SQ tree SLIT‐tablet reduce ARC symptoms triggered by birch or oak pollen. The optimal dose for further development was 12 DU. Clinical and immunological findings suggest that the tablet may be used to treat allergies to all species within the birch homologous group.

Communities of practice (CoPs) have been adopted by several industries, in particular healthcare, and researchers have proposed explanatory frameworks and guidelines for development. However, first‐line managers are left with sparse research‐based advice on how to develop CoP due to a lack of empirical studies from operations. In response, this paper proposes and tests a framework for intentional development of CoPs in healthcare operations. The proposed framework is structured in six steps, which incorporate seven factors for CoP development as identified in the literature. A double case study test demonstrates that the proposed framework can be applied to develop CoPs within operations, resulting in knowledge sharing between CoP participants, creation of new knowledge, and improvement of practice. In particular, the tests find the following steps to be especially crucial for CoP development: An introduction to the manager of the functions and benefits of CoPs to ensure resource allocation (e.g., time to participate and a meeting room), choosing a practice with relevance for manager and employees, appointing a coordinator, and inviting employees to participate voluntarily. This paper contributes to CoP theory with explanations of how and why specific actions develop CoP within operations. The contribution to practitioners is a step‐by‐step framework to intentionally develop CoPs within operations to improve knowledge sharing and practice improvement.

Nicotine exposure during gestation is associated with a higher risk of adverse behavioral outcomes including a heightened liability for dependency to drugs of abuse, which can exhibit drug-specificity influenced by gender. This enhanced liability suggests that nicotine use during pregnancy alters neural development in circuits involved in motivation and reward-based learning. The ventral tegmental area (VTA) is critical in motivated behaviors and we hypothesized that gestational exposure to nicotine alters the development of excitatory circuits in this nucleus. Accordingly, in VTA brain slices from male and female mice exposed to nicotine during the prenatal period (PNE) and controls, we compared cellular rises in calcium induced by AMPA receptor and nicotinic acetylcholine receptor (nAChR) stimulation by use of the ratiometric calcium binding dye, Fura-2AM. We found that AMPA induced smaller amplitude calcium rises in the PNE VTA, which was an effect only detected in males. Further, while the amplitude did not vary between treatment and control in females, a greater number of cells responded with rises in calcium in the PNE. Conversely, the proportions of cells responding with calcium rises induced by nAChR stimulation did not change in either gender according to treatment. However, larger rises in calcium in PNE females were detected. When taken together our data show that excitatory signaling in the VTA is altered in a gender-specific manner by PNE and suggest that alterations in signaling could play a role in drug-specific differences in maladaptive, motivated behaviors exhibited by males and females born to mothers exposed to nicotine during pregnancy. © 2018 Wiley Periodicals, Inc. Develop Neurobiol 2018.

Background Disability prevalence estimates are used to identify populations, establish priorities and allocate funding for a broad range of federal, state, and local initiatives. Increasingly, these estimates are based on a set of six questions developed and tested for use in the American Community Survey (ACS). A key assumption about the ACS disability screeners is that they sufficiently capture the entire population of people with disabilities, but some studies indicate that certain disability groups are underrepresented. Objectives The objective of this study is to explore potential underrepresentation of certain disability groups identified by the ACS disability questions. Methods We compared disability prevalence rates from two data sources for adolescents with disabilities, aged 14 to 16, who receive Supplemental Security Income (SSI, n = 2051). The Social Security Administration (SSA) provided disability determination data for each adolescent, and adolescents (or proxy-rater) provided baseline self-report data about functional limitation based on the six ACS disability questions. Results Approximately 17% of the sample did not endorse any ACS questions. Excluding SSA categories with cell counts less than 10, the top five conditions not captured by ACS questions included respiratory conditions (38%), mood disorders (28%), other mental disorders (27%), schizophrenic and other psychotic disorders (27%) and developmental disorders (20%). Conclusions Our findings suggest that people with mental health and developmental disabilities and those with respiratory conditions are among those groups under-represented by the ACS disability questions. Changes or additions to ACS questions should be considered to ensure that all disability groups are addressed in public health planning.

A juvenile, wild-caught prairie falcon ( Falco mexicanus) kept for falconry was presented to a veterinary hospital for intermittent opisthotonos and torticollis. Clinical examination, complete blood count, serum biochemistry panel, and fecal analysis were unremarkable. Clinical signs did not resolve, and the bird was euthanized 6 mo after the appearance of clinical signs. Autopsy revealed a mild, unilateral hydrocephalus and nematodes within the thoracic air sac. Histopathology demonstrated mild, unilateral hydrocephalus; scattered glial nodules; meningeal nematode sections; and meningeal and intraventricular embryonated eggs. Morphology and molecular characterization were consistent with the air sac nematode Serratospiculum or Serratospiculoides spp. Air sac nematode infection can be associated with air sacculitis or pneumonia in falcons. Aberrant migration of air sac filariid nematodes Serratospiculum or Serratospiculoides spp. into the nervous system resulting in clinical disease is rare, but should be included in the differential diagnosis of neurologic diseases in falcons.

Transglutaminase (TG) catalyzes formation of covalent bonds between lysine and glutamine side chains and has applications in manipulation of food structure. Physical properties of a whey protein mixture (SPC) denatured either at elevated pH or by heat-treatment and followed by TG catalyzed crosslinking, have been characterised using dynamic light scattering, size exclusion chromatography, flourescence spectroscopy and atomic force microscopy. The degree of enzymatic crosslinking appeared higher for pH- than for heat-denatured SPC. The hydrophobic surface properties depended on the treatment, thus heating caused the largest exposure of the hydrophobic core of SPC proteins, which was decreased by crosslinking. The particle size of the treated SPC samples increased upon crosslinking by TG. Moreover, the particle morphology depended on the type of denaturing treatment, thus heat-treated SPC contained fibrillar structures, while pH-denatured SPC remained globular as documented by using atomic force microscopy. Finally, the in vitro digestability of the different SPC samples was assessed under simulated gastric and intestinal conditions. Notably heat-treatment was found to lower the gastric digestion rate and enzymatic crosslinking reduced both the gastric and the intestinal rate of digestion. These characteristics of the various SPC samples provide a useful basis for design of isoenergic model foods applicable in animal and human studies on how food structure affects satiety.

Interactions of exopolysaccharides and proteins are of great importance in food science, but complicated to analyze and quantify at the molecular level. A surface plasmon resonance procedure was established to characterize binding of seven structure-determined, branched hetero-exopolysaccharides (HePSs) of 0.14–4.9 MDa from lactic acid bacteria to different milk proteins (β-casein, κ-casein, native and heat-treated β-lactoglobulin) at pH 4.0–5.0. Maximum binding capacity (RUmax) and apparent affinity (KA,app) were HePS- and protein-dependent and varied for example 10- and 600-fold, respectively, in the complexation with native β-lactoglobulin at pH 4.0. Highest RUmax and KA,app were obtained with heat-treated β-lactoglobulin and β-casein, respectively. Overall, RUmax and KA,app decreased 6- and 20-fold, respectively, with increasing pH from 4.0 to 5.0. KA,app was influenced by ionic strength and temperature, indicating that polar interactions stabilize HePS–protein complexes. HePS size as well as oligosaccharide repeat structure, conferring chain flexibility and hydrogen bonding potential, influence the KA,app.

Experimental and theoretical studies on ion-lipid interactions, predict that binding of calcium ions to cell membranes leads to macroscopic mechanical effects and membrane remodeling. Herein, we provide experimental evidence that a point-source of Ca2+ acting upon a negatively charged membrane, generates spontaneous curvature and triggers the formation of tubular protrusions that point away from the ion source. This behavior is rationalized by strong binding of the divalent cations to the surface of the charged bilayer which effectively neutralizes the surface charge density of outer leaflet of the bilayer. The mismatch in the surface charge density of the two leaflets, leads to non-zero spontaneous curvature. We probe this mismatch through the use of molecular dynamics simulations, and validate that calcium ion binding to a lipid membrane is sufficient to generate inward spontaneous curvature, bending the membrane. Additionally, we demonstrate that the formed tubular protrusions can be translated along the vesicle surface in a controlled manner by repositioning the site of localized Ca2+ exposure. The findings demonstrate lipid membrane remodeling in response to local chemical gradients and offer potential insights into the cell membrane behavior under conditions of varying calcium ion concentrations.

The microbial communities in spontaneously fermented camel milk from Ethiopia were characterised through metagenomic 16S rRNA sequencing and lactic acid bacteria were isolated with the goal of selecting strains suitable as starter cultures. The fermented camel milk microbiota was dominated either by Lactobacillales or by Enterobacteriaceae, depending on incubation temperature and the provider of the milk. Strains of species with a potential use as starter cultures i.e., Lactococcus lactis, Lactobacillus plantarum, and Pediococcus acidilactici, were isolated. Fast acidifiers of camel milk have been isolated from the species of Lc. lactis, P. acidilactici, and Streptococcus infantarius. Gram-negative and potentially pathogenic microorganisms were frequent in spontaneously fermented camel milk, indicating the need for improved hygiene in Ethiopian camel farms. The profiled microbiota of spontaneously fermented camel milk and the isolated LAB strains will significantly contribute towards improving food safety and food security in dry regions that depend on camel milk production.

The aim of the study was to investigate how microparticulated and nanoparticulated whey proteins mixed with alginate respond to simulated in vitro gastric digestion conditions at pH 3.0. Initially, particle size distributions and zeta potential were measured in all mixtures at pH 3.0. Particle size distributions as well as SDS-PAGE were used to investigate the rate of protein degradation by pepsin during simulated in vitro gastric digestion. The complexation of nanoparticulated and microparticulated whey protein with alginates causes formation of insoluble and soluble complexes, which can resist pepsin degradation to a different degree. These results highlight the potential of developing new food products, which can enhance satiety.

Ultrasound is being increasingly investigated for real-time target localization in image-guided interventions. Yet, in-vivo validation remains challenging due to the difficulty to obtain a reliable ground truth. For this purpose, real-time volumetric (4D) ultrasound imaging was performed simultaneously with electromagnetic localization of three wireless transponders implanted in the liver of a radiotherapy patient. 4D ultrasound and electromagnetic tracking were acquired at framerates of 12Hz and 8Hz, respectively, during free breathing over 8 min following treatment. The electromagnetic antenna was placed directly above and the ultrasound probe on the right side of the patient to visualize the liver transponders. It was possible to record 25.7 s of overlapping ultrasound and electromagnetic position data of one transponder. Good spatial alignment with 0.6 mm 3D root-mean-square error between both traces was achieved using a rigid landmark transform. However, data acquisition was impaired since the electromagnetic tracking highly influenced the ultrasound equipment and vice versa. High intensity noise streaks appeared in the ultrasound scan lines irrespective of the chosen frequency (1.7-3.3 MHz, 2/4 MHz harmonic). To allow for target visualization and tracking in the ultrasound volumes despite the artefacts, an online filter was designed where corrupted pixels in the newest ultrasound frame were replaced with non-corrupted pixels from preceding frames. Aside from these artefacts, the recorded electromagnetic tracking data was fragmented and only the transponder closest to the antenna could be detected over a limited period of six consecutive breathing cycles. This problem was most likely caused by interference from the metal holder of the ultrasound probe and was solved in a subsequent experiment using a 3D-printed non-metal probe fixation. Real-time wireless electromagnetic tracking was compared with 4D ultrasound imaging in-vivo for the first time. For stable tracking, large metal components need to be avoided during data acquisition and ultrasound filtering is required.

Given vocational rehabilitation’s (VR) substantial role in workforce development, it follows that agencies would encourage and support consumers using the most current job-seeking methods, including social media. Recent data, however, show that online strategies are limited in VR practice and that many agencies lack comprehensive policies to guide social media use. We conducted a qualitative analysis to examine current VR social media policies and procedures and offer recommendations for policy development. Through an examination of open coding of 22 policies and one guideline, we identified seven themes and 25 subthemes, which mapped onto recommended social media policy elements from research. Only two VR policies, however, covered themes in a comprehensive way, and only one policy acknowledged social media as a tool for consumer job search. This outcome is an oversight given the emphasis social media plays in employment networking, proof of performance, and skills demonstration when seeking employment.

The bacterial Shiga toxin interacts with its cellular receptor, the glycosphingolipid globotriaosylceramide (Gb3 or CD77), as a first step to entering target cells. Previous studies have shown that toxin molecules cluster on the plasma membrane, despite the apparent lack of direct interactions between them. The precise mechanism by which this clustering occurs remains poorly defined. Here, we used vesicle and cell systems and computer simulations to show that line tension due to curvature, height, or compositional mismatch, and lipid or solvent depletion cannot drive the clustering of Shiga toxin molecules. By contrast, in coarse-grained computer simulations, a correlation was found between clustering and toxin nanoparticle-driven suppression of membrane fluctuations, and experimentally we observed that clustering required the toxin molecules to be tightly bound to the membrane surface. The most likely interpretation of these findings is that a membrane fluctuation-induced force generates an effective attraction between toxin molecules. Such force would be of similar strength to the electrostatic force at separations around 1 nm, remain strong at distances up to the size of toxin molecules (several nanometers), and persist even beyond. This force is predicted to operate between manufactured nanoparticles providing they are sufficiently rigid and tightly bound to the plasma membrane, thereby suggesting a route for the targeting of nanoparticles to cells for biomedical applications.

Purpose To assess the outcomes of treatment with a dermal regeneration template (DRT) in a cohort of combat casualties with severe upper extremity injuries. Methods Records of all active duty military patients treated with DRT at our institution between November 2009 and July 2013 were screened. Inclusion criteria were upper extremity open wounds sustained during combat, requiring split-thickness or full-thickness skin grafting for closure. The primary outcome measure was wound healing after the first attempt at definitive treatment (defined as the first application of split-thickness or full-thickness skin graft). Independent variables collected included time from injury to arrival at our facility, mechanism of injury, wound infection, tobacco use, location of wound, number of operative debridements, and patient demographics. Results A total of 60 patients with 69 wounds met the inclusion criteria. Most wounds were to the wrist or forearm (54%) or fingers (19%). All wounds were heavily contaminated, requiring a mean of 2.5 operative debridements before DRT placement. All wounds treated with full-thickness skin grafting after DRT healed completely without further complication. Split-thickness skin grafting was successful in 96% of patients. Conclusions DRT wound dressings are a helpful adjunct in the treatment of contaminated war wounds to the upper extremity. Type of study/level of evidence Therapeutic IV.

The structure of functional lipid domains (rafts) in biological membranes has for long time been unresolved due to their small length scales and transient nature. These cooperative properties of the lipid bilayer matrix are modelled by free-standing giant unilammellar vesicles (GUVs) with well-characterized lipid composition. We review a series of recent advances in preparation and analysis of GUVs, which allows for characterization of small domains by high-resolution imaging techniques. These includes a new GUV preparation method with a desired overall lipid composition achieved by mixing small unilammellar vesicles (SUVs), test of the lipids compositional uniformity in GUVs and swift adsorption of GUVs to solid support by kinetically arresting the lateral structure of membrane prior to collapse for subsequent imaging. The techniques are applied to the analysis of membrane domains in GUVs formed from mixtures of DOPC/DPPC/cholesterol with and without Na,K-ATPase (NKA), a transmembrane protein known to be associated with rafts. Two mechanisms of domain formation are revealed: 1) close to lo/ld phase coexistence, domains in size up to 100 nm appear as thermally induced droplet fluctuations, 2) NKA shows interfacial activity and cluster in lo/ld micro-emulsion droplets. Some perspectives for the application of the techniques and the understanding of the nature of raft domains are outlined.

BACKGROUND: Results-based funding models can improve Vocational Rehabilitation (VR) client employment outcomes, but evidence suggests that gaps in services occur when provider risk is not compensated. OBJECTIVE: The purpose of this qualitative study is to learn how VR agencies deliver and pay for job development and placement services, the factors that shape decision-making, and the outcomes of such decisions. METHODS: Administrators from 40 VR agencies identified informants to take part in qualitative interviews about how their agency delivers, contracts, and pays for job development services. Interview notes were analyzed to identify themes and dimensions in the data. RESULTS: Gaps in provider services exist in rural communities, where provider risk is increased due to economic, transportation, and delivery barriers. Strategies to overcome barriers focused on expanding available providers by lowering requirement for provider entry, increasing recruitment activities by the VR agency, reducing financial risk in the form of incentives, tiered payments, and shared funding models, and increasing agency capacity and support. CONCLUSIONS: Strategies for effective VR service delivery are shaped by multiple factors and choice points at the agency level. A deeper understanding of the intended and unintended consequences of service delivery models is an important contribution to informed decision-making.

In rodents, prenatal nicotine exposure (PNE) has been associated with increased risk for development of cognitive and emotional disturbances, but the findings are somewhat conflicting. Lack of behavioral alterations following PNE could be due to the variety of methods available for nicotine delivery, exposure time and species used, with inbred strains being mostly employed. Such differences suggest the need to investigate the behavioral phenotype in each PNE model available if we are to find models with enhanced translational value. In this study, we assessed sex-dependent effects of PNE on ADHD-related behaviors and on the levels of mRNA coding for glutamate receptor subunits within the prefrontal cortex in the outbred NMRI mice exposed to nicotine via maternal drinking water during gestation. Cotinine levels were assessed in newborn pups. Behaviors related to anxiety, compulsivity, working memory, and locomotion were evaluated in both sexes of young adult offspring using the elevated zero maze, marble burying, spontaneous alternation behavior, and locomotor activity tests. Expression of mRNA coding for different glutamate receptors subunits within the prefrontal cortex (PFC) was measured using RT-qPCR. Cotinine levels in the serum of newborns confirmed fetal nicotine exposure. Both male and female offspring showed ADHD-like behaviors, such as deficit in the SAB test and hyperactivity. In addition, PNE male mice displayed anxiety- and compulsive-like behaviors, effects that were absent in female offspring. Finally, PNE reduced the mRNA expression of GluN1-, GluN2B-, and mGluR2-related genes within the PFC of male offspring, whereas it reduced the expression of mRNA coding for GluA2 subunit in female mice. PNE in NMRI mice induced sex-dependent behavioral changes, which parallels clinical findings following maternal cigarette smoke exposure. Alterations detected in PFC mRNA glutamate receptor proteins could contribute to the abnormal behavioral responses observed, but other signaling pathways or brain regions are likely involved in the behavioral susceptibility of PNE individuals.

ObjectiveTo evaluate the relative risk (RR) of serious and non-serious adverse events in patients treated with exercise therapy compared with those in a non-exercising control group.DesignSystematic review and meta-analysis.Data sourcesPrimary studies were identified based on The Cochrane Database of Systematic Reviews investigating the effect of exercise therapy.Eligibility criteriaAt least two of the authors independently evaluated all identified reviews and primary studies. Randomised controlled trials were included if they compared any exercise therapy intervention with a non-exercising control. Two authors independently extracted data. The RR of serious and non-serious adverse events was estimated separately.Results180 Cochrane reviews were included and from these, 773 primary studies were identified. Of these, 378 studies (n=38 368 participants) reported serious adverse events and 375 studies (n=38 517 participants) reported non-serious adverse events. We found no increase in risk of serious adverse events (RR=0.96 (95%CI 0.90 to 1.02, I2: 0.0%) due to exercise therapy. There was, however, an increase in non-serious adverse events (RR=1.19 (95%CI 1.09 to 1.30, I2: 0.0%). The number needed to treat for an additional harmful outcome for non-serious adverse events was 6 [95%CI 4 to 11).ConclusionParticipating in an exercise intervention increased the relative risk of non-serious adverse events, but not of serious adverse events. Exercise therapy may therefore be recommended as a relatively safe intervention.PROSPERO registration numberCRD42014014819.

We present a multi-scale simulation procedure to describe membrane-related biological processes that span over a wide range of length scales. At macroscopic length-scale, a membrane is described as a flexible thin film modeled by a dynamic triangulated surface with its spatial conformations governed by an elastic energy containing only a few model parameters. An implicit protein model allows us to include complex effects of membrane-protein interactions in the macroscopic description. The gist of this multi-scale approach is a scheme to calibrate the implicit protein model using finer scale simulation techniques e.g., all atom and coarse grain molecular dynamics. We previously used this approach and properly described the formation of membrane tubular invaginations upon binding of B-subunit of Shiga toxin. Here, we provide a perspective of our multi-scale approach, summarizing its main features and sketching possible routes for future development.

BACKGROUND: Social isolation has been compared to smoking in terms of risk to public health. Some groups are at particularly high risk for these feelings, including people with disabilities and rural residents. Few studies have considered the potentially compounding effects of disability status and rural residency. OBJECTIVE: To evaluate how reported satisfaction with social participation and perceived isolation relate to the health of rural and urban people with disabilities, and to consider whether number of disabilities, living arrangement, and employment status were associated with differences in reported satisfaction with social participation and perceived isolation. METHODS: This observational, cross-sectional analysis utilized data from working-age adults with disabilities (n = 1246) collected by the Collaborative on Health Reform and Independent Living (CHRIL). RESULTS: There were significant associations between reported health and measures of satisfaction with social participation and perceived isolation (all ps < .001). Increased number of disability issues, not being employed, and living with at least one other person were associated with reduced satisfaction with social participation (ps < .01), and number of disability issues and not being employed were associated with increased perceived isolation (ps < .01). Urban residents reported feeling more isolated (ps < .05) and there were multiple predictor x geographic residency (rural versus urban) interactions. CONCLUSION: These results underscore the importance of considering geography as a factor in understanding satisfaction with social participation and perceived isolation and how these factors relate to health in people with disabilities.

This study was conducted to evaluate the safety and bacterial profile of Dhanaan (Ethiopian traditional fermented camel milk). The composition of the microbial community in Dhanaan samples was analysed by a metagenomic approach of 16S rRNA gene amplicon sequencing. Metagenomic profiling identified 87 different bacterial microorganisms (OTUs) in six samples analysed. Although the Dhanaan samples contained various lactic acid bacteria (LAB), they also all contained undesirable microorganisms in large proportions. The following LAB genera were identified: Streptococcus, Lactococcus and Weissella. One Streptococcus species represented by OTU-1 (operational taxonomic unit) was found in all Dhanaan samples and the dominating species in four out of six samples. This common isolate was found to be closely related to S. lutetiensis and S. infantarius. Undesirable microorganisms from genera such as Escherichia, Klebsiella, Enterobacter, Acinetobacter and Clostridium were, however, also frequent, or even dominant in Dhanaan samples. Thus, this calls for a change in the Dahnaan manufacturing practice to an improved and safer production system. Starter cultures suitable for Dhanaan production might be developed from the Streptococcus, Weissella and Lactococcus microorganisms identified in this study. However, further safety evaluation and technological characterization need to be conducted on strains defined by OTU-1, OTU-2, OTU-3, OTU-8 and OTU-35 before they can be used as food grade starter cultures.

Background: Preoperative staging with 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) combined with computed tomography (CT) is used for the evaluation of metastatic disease in patients with invasive bladder cancer. The use of quantification with maximum standardized uptake value (SUVmax) of regional lymph nodes (LNs) has been suggested to increase the diagnostic ability for detection of malignancy. Objective: Assessment of the utility and clinical relevance of SUVmax in 18F-FDG PET in detecting regional nodal metastases in patients considered for radical cystectomy. Design, setting, and participants: From 2011 to 2014, we identified a total of 119 patients with urothelial carcinoma who underwent radical cystectomy with extended LN dissection; additionally, 12 patients were identified by preoperative biopsy. All patients underwent 18F-FDG PET/CT before treatment recommendation. Pathological findings were compared with preoperative PET/CT staging and analysed in a regional- or patient-based model according to SUVmax values. Outcome measurements and statistical analysis: In total, 2291 LNs were identified in 131 patients; locoregional involvement of 85 LNs were confirmed in 34 patients. Results and limitations: SUVmax >2 analysis: sensitivity. ±. 95% confidence interval of 79.4% (62.1-91.3) and specificity 66.5% (55.7-75.3). SUVmax >4 based analysis: sensitivity was 61.8% (43.6-77.8) and specificity was 84.5% (75.8-91.1). Two years of follow-up implied that higher SUVmax is correlated with higher recurrence risk, independent of conventional pathological findings. Conclusions: 18F-FDG PET/CT using SUVmax of LNs is a useful tool for preoperative evaluation of pelvic LN metastases from invasive bladder cancer and contributes to the selection of patients for personalized treatment. Patient summary: In this report, we establish that it is possible to identify disease from bladder cancer in the lymphatic tissue surrounding the bladder using a scan analysis. This assists in the selection of treatment for patients with bladder cancer and may spare patients from unnecessary procedures. In this report, we found that 18F-fluorodeoxyglucose positron emission tomography/computed tomography is able to identify metastatic disease in regional lymph nodes in bladder cancer patients. The use of maximum standardized uptake values improve the evaluation of lymph nodes prior to treatment.

Real-time volumetric (4D) ultrasound has shown high potential for diagnostic and therapy guidance tasks. One of the main drawbacks of ultrasound imaging to date is the reliance on manual probe positioning and the resulting user dependence. Robotic assistance could help overcome this issue and facilitate the acquisition of long-term image data to observe dynamic processes in vivo over time. The aim of this study is to assess the feasibility of robotic probe manipulation and organ motion quantification during extended imaging sessions. The system consists of a collaborative robot and a 4D ultrasound system providing real-time data access. Five healthy volunteers received liver and prostate scans during free breathing over 30 min. Initial probe placement was performed with real-time remote control with a predefined contact force of 10 N. During scan acquisition, the probe position was continuously adjusted to the body surface motion using impedance control. Ultrasound volumes, the pose of the end-effector and the estimated contact forces were recorded. For motion analysis, one anatomical landmark was manually annotated in a subset of ultrasound frames for each experiment. Probe contact was uninterrupted over the entire scan duration in all ten sessions. Organ drift and imaging artefacts were successfully compensated using remote control. The median contact force along the probe’s longitudinal axis was 10.0 N with maximum values of 13.2 and 21.3 N for liver and prostate, respectively. Forces exceeding 11 N only occurred in 0.3% of the time. Probe and landmark motion were more pronounced in the liver, with median interquartile ranges of 1.5 and 9.6 mm, compared to 0.6 and 2.7 mm in the prostate. The results show that robotic ultrasound imaging with dynamic force control can be used for stable, long-term imaging of anatomical regions affected by motion. The system facilitates the acquisition of 4D image data in vivo over extended scanning periods for the first time and holds the potential to be used for motion monitoring for therapy guidance as well as diagnostic tasks.

Background Trust of information shapes adherence to recommended practices and speed of public compliance during public health crises. This is particularly important for groups with higher rates of high-risk health conditions, including those aged 65 and over and people with disabilities. Objective We examined trust in information sources and associated adherence to COVID-19 public health recommendations among people with disabilities living in metropolitan, micropolitan, and noncore counties. Methods We recruited participants using Amazon’s Mechanical Turk (MTurk) and screened for disability status (n = 408). We compared sociodemographic groups with t-tests, Pearson’s correlations, and Chi-square, as appropriate. We used linear regression to examine factors associated with trust in information and compliance with CDC recommended COVID-19 practices. Results Nonmetro respondents had the lowest trust ratings among all demographic groups, and reported significantly less trust in most information sources. Respondents aged 65 and over reported the highest compliance with CDC recommended practices, while those from nonmetro areas reported the lowest. A regression model for adherence to CDC recommended practices was significant (F = 11.87, P ≤ .001), and explained 33% of the variance. Specifically, increased adherence was associated with being over 65, female, and higher general trust scores. Decreased practices were associated with being nonwhite, nonmetro, higher trust scores in President Trump, and having a communication disability. Conclusions Trust in information sources is associated with action. It is important to provide clear, consistent, and non-polarizing messages during public health emergencies to promote widespread community action.