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Background: The alpha-emitting radionuclide therapy [223Ra]RaCl2 (Radium-223) improves overall survival (OS) and time to symptomatic skeletal event (SSE) in patients with metastatic castration-resistant prostate cancer (mCRPC). Evidence suggests that the effect of Radium-223 is partly exerted through an impact on the surrounding bone matrix. We hypothesized that bone metabolism markers (BMM) could provide predictive information regarding response to Radium-223. Accordingly, the aim of this study was to investigate changes in BMM during Radium-223 therapy and evaluate association with clinical outcome. Methods: Prospective study of BMM in patients with mCRPC receiving Radium-223. Blood samples were collected before each administration of Radium-223 and the following BMM were quantified; bone-specific alkaline phosphatase (BALP), osteocalcin, procollagen type I N-propeptide (PINP), C-terminal telopeptide of type I collagen (CTX), C-terminal cross-linking telopeptide of type I collagen generated by matrix metalloproteinases (CTX-MMP), tartrate-resistant acid phosphatase isoform 5b (TRACP5b), receptor-activated nuclear factor κB ligand (RANKL), osteoprotegerin (OPG), and sclerostin. Clinical outcomes were scintigraphic progression during/after therapy, change in bone scan index (BSI), occurrence of SSE, and OS. Results: A total of 55 mCRPC patients were included. There was a significant linear association between skeletal extent of disease and CTX-MMP, PINP, BALP, and osteocalcin. No significant association between dynamics in BSI and BMM were detected. Median OS for the cohort was 14 months (95% CI: 10.7–16.8). Baseline levels of Log2-CTX-MMP (HR = 2.15 (95%CI: 1.1–4.1)) and Log2-BALP (HR = 1.59 (95%CI: 1.1–2.1)) were associated with OS. Patients with increasing CTX-MMP during therapy had significantly shorter OS (Median OS = 4 mo. (95%CI: 2.3–5.7)) than patients with stable or decreasing CTX-MMP (Median OS = 12 mo. (95%CI: 10.1–13.9), P < 0.001). Conclusion: BMM are significantly associated with scintigraphic extent of skeletal disease and OS in patients with mCRPC. Particularly, the bone resorption marker CTX-MMP is a promising surrogate marker for prediction of outcome in patients receiving Radium-223 therapy and could potentially improve selection of patients for therapy and assessment of response. Trial registration: Clinicaltrials.gov, NCT03247010. Registered 10th of August 2017, https://clinicaltrials.gov/study/NCT03247010?term=NCT03247010&rank=1. [ABSTRACT FROM AUTHOR]

A pivotal animal model for development of anticancer molecules is mice with subcutaneous tumors, grown by injection of xenografted tumor cells, where micro-Computed Tomography (µCT) of the mice is used to analyze the efficacy of the anticancer molecule. Manual delineation of the tumor region is necessary for the analysis, which is time-consuming and inconsistent, highlighting the need for automatic segmentation (AS) tools. This study introduces a preclinical µCT database, comprising 452 whole-body scans from 223 individual mice with subcutaneous tumors, spanning ten diverse µCT datasets conducted between 2014 and 2020 on a preclinical PET/CT scanner, making it the hitherto largest dataset of its kind. Each tumor is annotated manually by three expert annotators, allowing for robust model development. Inter-annotator agreement was analyzed, and we report an overall annotation agreement of 0.903 ± 0.046 (mean ± std) Fleiss' Kappa and a mean deviation in volume estimation of 0.015 ± 0.010 cm3 (6.9% ± 4.7), which establishes a human baseline accuracy for delineation of subcutaneous tumors, while showing good inter-annotator agreement. [ABSTRACT FROM AUTHOR]

Purpose: Assessing work functioning in patients with persistent low back pain (LBP) is important for understanding their ability to engage in work-related activities. This study aims to evaluate the item characteristics, factor structure, and internal consistency of the Work Rehabilitation Questionnaire (WORQ) in patients with persistent LBP. Methods: Four hundred and twenty-five individuals with LBP completed the WORQ. Item characteristics, exploratory factor analysis (EFA), and consistency were performed to identify the underlying factors. Results: Missing responses were < 2% for each item. The analysis revealed three factors: psychological wellbeing, physical functioning, and cognitive ability. The factors demonstrated strong internal consistency, with Cronbach's alpha values ranging from 0.88 to 0.93 and McDonald's Omega from 0.92 to 0.96. Fifteen items did not fit into any identified factors, suggesting their potential value in screening functioning levels beyond the factors. Conclusions: The WORQ is a valid instrument for evaluating work limitations in individuals with persistent LBP. Further research should assess its responsiveness to changes from interventions that target workability. Advancing this knowledge has the potential to promote work rehabilitation and improve the quality of life for patients with persistent LBP. [ABSTRACT FROM AUTHOR]

Background: Optimal balance between macro- and microcirculation in critically ill patients is crucial for ensuring optimal organ perfusion. Nitric oxide (NO) is a regulator of vascular hemostasis and tone. The availability of NO is controlled by asymmetric dimethylarginine (ADMA), symmetric dimethylarginine (SDMA), and the availability of the NO substrates arginine and homoarginine. We investigated the changes in plasma concentrations of ADMA, SDMA, arginine, and homoarginine days 1–5 of intensive care unit (ICU) admission and the association between the change in concentration days 1–3 and 30-day all-cause mortality. Methods: Single-center cohort study of adult critically ill patients from the ICU at Copenhagen University Hospital – North Zealand. ADMA, SDMA, arginine, and homoarginine (NO-biomarkers) were measured on days 1–5. Initially, we determined the changes in NO-biomarkers days 1–5 with linear mixed models, and subsequently how the changes in NO-biomarkers days 1–3 were associated with 30-day all-cause mortality. Post-hoc we analyzed the association between plasma concentration at admission and 30-day all-cause mortality. Results: In total 567 out of 577 patients had plasma samples from days 1–5. Plasma concentrations of ADMA and arginine increased from days 1–5. SDMA concentrations increased from days 1–2, followed by a decrease from days 2–5. Concentrations of homoarginine did not change from days 1–3 but slightly increased from days 3–5. In total 512 patients were alive 3 days after ICU admission. Among these patients, a daily twofold increase in ADMA concentration from days 1–3 was associated with decreased mortality in multivariate analysis (HR 0.45; 95% CI 0.21–0.98; p = 0.046). An increase in SDMA, arginine, or homoarginine was not associated with mortality. Post-hoc we found that a twofold increase in ADMA or SDMA concentrations at admission was associated with mortality (HR 1.78; 95% CI 1.24–2.57; p = 0.0025, and HR 1.41; 95% CI 1.05–1.90; p = 0.024, respectively). Conclusions: Increasing ADMA concentrations on days 1–3 are inversely associated with mortality, however not with the same strength as high ADMA or SDMA concentrations at admission. We suggest that admission concentrations are the focus of future research on ADMA and SDMA as predictors of mortality or potential therapeutical targets in ICU patients. [ABSTRACT FROM AUTHOR]

Engineered DNA will slow the growth of a host cell if it redirects limiting resources or otherwise interferes with homeostasis. Escape mutants that alleviate this burden can rapidly evolve and take over cell populations, making genetic engineering less reliable and predictable. Synthetic biologists often use genetic parts encoded on plasmids, but their burden is rarely characterized. We measured how 301 BioBrick plasmids affected Escherichia coli growth and found that 59 (19.6%) were burdensome, primarily because they depleted the limited gene expression resources of host cells. Overall, no BioBricks reduced the growth rate of E. coli by >45%, which agreed with a population genetic model that predicts such plasmids should be unclonable. We made this model available online for education (https://barricklab.org/burden-model) and added our burden measurements to the iGEM Registry. Our results establish a fundamental limit on what DNA constructs and genetic modifications can be successfully engineered into cells. Engineered DNA will slow the growth of a host cell if it redirects limiting resources or otherwise interferes with homeostasis. Here the authors measure how 301 BioBrick plasmids affected Escherichia coli growth and found that 19.6% were burdensome, primarily because they depleted the limited gene expression resources of host cells. [ABSTRACT FROM AUTHOR]

Yellow rust caused by Puccinia striiformis f. sp. tritici (Pst) is one of the most important wheat diseases. Adult plant resistance (APR) genes have gained the attention of breeders and scientists because they show higher durability compared to major race-specific genes. Here, we determined the effect of the APR genes Yr18, Yr29 and Yr46 in North-West European field conditions against three currently important Pst races. We used three pairs of sibling wheat lines developed at CIMMYT, which consisted of a line with the functional resistance gene and a sibling with its non-functional allele. All APR genes showed significant effects against the Pst races Warrior and Warrior (–), and a race of the highly aggressive strain PstS2. The effects of Yr18 and Yr46 were especially substantial in slowing down disease progress. This effect was apparent in both Denmark, where susceptible controls reached 100 percent disease severity, and in United Kingdom where disease pressure was lower. We further validated field results by quantifying fungal biomass in leaf samples and by micro-phenotyping of samples collected during early disease development. Microscopic image analyses using deep learning allowed us to quantify separately the APR effects on leaf colonization and pustule formation. Our results show that the three APR genes can be used in breeding yellow rust resistant varieties of spring wheat to be grown in North-West European conditions, and that deep learning image analysis can be an effective method to quantify effects of APR on colonisation and pustule formation. [ABSTRACT FROM AUTHOR]

Background: Bullous pemphigoid (BP) is a rare, autoimmune, blistering skin disease associated with high disease burden, profoundly decreased quality of life and increased morbidity. Emerging evidence supports an important role for type 2 inflammation in disease pathogenesis. Current management relies on topical and/or systemic corticosteroids, non-selective immunosuppressants and antibiotics with anti-inflammatory properties, which are all limited by side effects and toxicities. Therefore, targeted, efficacious and safe therapies are needed. Dupilumab blocks the shared receptor component for interleukin (IL)-4 and IL-13, key and central drivers of type 2 inflammation. Several reports of patients successfully treated with dupilumab have been published; however, dupilumab has not been formally assessed in a double-blind, placebo-controlled trial. Objectives: We report the design of LIBERTY-BP ADEPT, a multicenter, randomized, double-blind, placebo-controlled clinical trial to evaluate the efficacy and safety of dupilumab in adults with BP. Methods: LIBERTY-BP ADEPT comprises a 35-day screening, 52-week treatment and 12-week follow-up period. Approximately 98 adults aged 18–90 years with moderate-to-severe BP are being enrolled at 51 sites on 4 continents and randomized 1:1 to subcutaneous dupilumab or placebo every 2 weeks. All participants will receive concomitant oral corticosteroids (OCS). Planned Outcomes: The primary endpoint is the proportion of patients achieving complete remission off steroid therapy at week 36. Key secondary endpoints include total cumulative OCS dose to week 36, percent change and proportion of patients with ≥ 4-point reduction in the weekly average of daily Peak Pruritus Numerical Rating Scale from baseline to week 36 and percent change in Bullous Pemphigoid Area Index score from baseline to week 36. Conclusion: The trial results will provide evidence on whether the efficacy and safety of dupilumab support its use as a potential novel treatment approach for BP and will provide new insights into the role of type 2 inflammation in BP pathogenesis. Clinical Trial Registration: ClinicalTrials.gov identifier NCT04206553. [ABSTRACT FROM AUTHOR]

Introduction: The standard surgical procedure for unstable ankle fractures is fixation of the lateral malleolus with a plate and screws. This method has a high risk of complications, especially among patients with fragile skin conditions. The aim of this study was to estimate the re-operation rates and identify complications in patients with an unstable ankle fracture, surgically treated with an intramedullary screw or rush pin. Materials and methods: We identified all patients who were surgically treated with either a 3.5-mm screw or rush pin at Aarhus University Hospital, Denmark, from 2012 to 2018. Major complications were re-operations within three months. We included 80 patients, of which 55 (69%) were treated with a 3.5-mm intramedullary screw and 25 (31%) with a rush pin. The majority of the study population was female (59) and the mean age was 75 (range 24 to 100) years. Of the 80 patients included, 41 patients had more than 2 comorbidities. Results: Three patients underwent re-operation within three months due to either fracture displacement or hardware cutout. Radiographs obtained after six weeks showed that nine patients had loss of reduction. Additionally, four patients had superficial wound infections and six patients had delayed wound healing. Conclusions: Intramedullary fixation of distal fibula fractures with either a screw or rush pin has low re-operation rates. However, the high proportion of patients with radiological loss of reduction is concerning. [ABSTRACT FROM AUTHOR]

Background and Objective: Preliminary evidence suggests a possible preventive effect of tumor necrosis factor-α inhibitors (TNFi) on incident dementia. The objective of the analysis was to investigate the association between TNFi and the risk of incident dementia in a population undergoing treatment for rheumatological disorders. Methods: We followed patients aged ≥ 65 years with dementia and rheumatological conditions in two cohort studies, DANBIO (N = 21,538), a Danish clinical database, and AOK PLUS (N = 7112), a German health insurance database. We defined incident dementia using diagnostic codes and/or medication use and used Cox regression to compare the associations of TNFi with other rheumatological therapies on the risk of dementia. To ensure that the patients were receiving long-term medication, we included patients with rheumatic diseases and systemic therapies. Results: We observed similar trends towards a lower risk of dementia associated with TNFi versus other anti-inflammatory agents in both cohorts (hazard ratios were 0.92 [95% confidence interval 0.76, 1.10] in DANBIO and 0.89 [95% confidence interval 0.63, 1.24] in AOK PLUS, respectively). Conclusions: Tumor necrosis factor-α inhibitors may decrease the risk of incident dementia although the association did not reach statistical significance in this analysis. Further research, ideally with randomization, is needed to gauge the potential of repurposing TNFi for dementia prevention and/or treatment. [ABSTRACT FROM AUTHOR]

Oxide inclusions appear in steel as a subproduct of steelmaking. These are generally detrimental to alloy properties; however, variations exist in the extent to which different inclusions are harmful because their properties vary as a function of their chemical composition. We use nanoindentation to measure the local elastic modulus and hardness of individual oxide particles, produced by precipitation within liquid iron, that belong to the systems Al2O3–SiO2–CaO and MnO–SiO2–FeO. Measured inclusion hardness values are typically in the range of 8 to 13 GPa and can reach 26 GPa for alumina-rich inclusions. Calcium aluminates rich in alumina are significantly stiffer than iron, with elastic moduli that can reach 350 GPa. On the contrary, calcium aluminates that are expected as a result of successful calcium treatment (i.e., with less than about 80 wt pct Al2O3 content) have elastic moduli below that of iron. This is also the case for the wide range of calcium aluminosilicates and of manganese silicates studied here. In addition, silicates containing about 70 to 80 wt pct MnO are observed to have a fine multiphase structure and an elastic modulus of ≈ 180 GPa. Those inclusions thus emerge as possible candidates if one aims to minimise, in loaded steel, stress concentrations associated with matrix-inclusion elastic mismatch. [ABSTRACT FROM AUTHOR]

Metalenses are flat lenses, where sub-wavelength, so-called meta-atoms manipulate the electric field to perform a given lens function. Compared to traditional lenses, the two main drawbacks of metalenses are their achromatic limitations and low efficiencies. While an abundance of simulations show that efficiencies above 90% are attainable for low numerical apertures (NA), experimental reports showing such high efficiencies are limited. Here, we use electron-beam lithography (EBL) to realize a set of lenses with varying NA from 0.08 to 0.93. The low NAs were expected to fit the model, and the higher NAs determine the validity range of the model. We find that measured efficiencies above 92% for NA = 0.24 are achievable, and that a slight modification of the simulation model extends its validility to NA = 0.6. Based on our results, we discuss that the lower efficiencies reported in the literature are caused by low-fidelity manufacturing, closing the efficiency gap between measurements and simulation in metalens fabrication. While numerical simulations for metalenses frequently show efficiencies above 90% at low numerical apertures, the experimental counterpart struggles to reach such efficiencies. The authors modify the model for prediction and systematically realise a set of high-precision meta-lenses with high efficiencies across the whole numerical aperture range. [ABSTRACT FROM AUTHOR]

The integrity of the intestinal mucus barrier is crucial for human health, as it serves as the body's first line of defense against pathogens. However, postnatal development of the mucus barrier and interactions between maturity and its ability to adapt to external challenges in neonatal infants remain unclear. In this study, we unveil a distinct developmental trajectory of the mucus barrier in preterm piglets, leading to enhanced mucus microstructure and reduced mucus diffusivity compared to term piglets. Notably, we found that necrotizing enterocolitis (NEC) is associated with increased mucus diffusivity of our large pathogen model compound, establishing a direct link between the NEC condition and the mucus barrier. Furthermore, we observed that addition of sodium decanoate had varying effects on mucus diffusivity depending on maturity and health state of the piglets. These findings demonstrate that regulatory mechanisms governing the neonatal mucosal barrier are highly complex and are influenced by age, maturity, and health conditions. Therefore, our results highlight the need for specific therapeutic strategies tailored to each neonatal period to ensure optimal gut health. [ABSTRACT FROM AUTHOR]

The lowest energy states in transition metal dichalcogenide (TMD) monolayers follow valley selection rules, which have attracted vast interest due to the possibility of encoding and processing of quantum information. However, these quantum states are strongly affected by temperature-dependent intervalley scattering leading to complete valley depolarization, which hampers practical applications at room temperature. Therefore, for achieving clear and robust valley polarization in TMD monolayers one needs to suppress parasitic depolarization processes, which is the central challenge in the growing field of valleytronics. Here, in electron-doping experiments on TMD monolayers, we show that strong doping levels beyond 1013 cm−2 can induce 61% and 37% valley contrast at room temperature in tungsten diselenide and molybdenum diselenide monolayers, respectively. Our findings demonstrate that charged excitons in TMD monolayers hold the potential for the development of efficient valleytronic devices functional at 300 K. [ABSTRACT FROM AUTHOR]

We here report the results of a mitral valve transcriptome study designed to identify genes and molecular pathways involved in development of congestive heart failure (CHF) following myxomatous mitral valve disease (MMVD) in dogs. The study is focused on a cohort of elderly age-matched dogs (n = 34, age ~ 10 years) from a single breed—Cavalier King Charles Spaniels (CKCS)—with a high incidence of MMVD. The cohort comprises 19 dogs (10♀, 9♂) without MMVD-associated CHF, and 15 dogs (6♀, 9♂) with CHF caused by MMVD; i.e., we compare gene expression in breed and age-matched groups of dogs, which only differ with respect to CHF status. We identify 56 genes, which are differentially expressed between the two groups. In this list of genes, we confirm an enrichment of genes related to the TNFβ-signaling pathway, extracellular matrix organization, vascular development, and endothelium damage, which also have been identified in previous studies. However, the genes with the greatest difference in expression between the two groups are CNTN3 and MYH1. Both genes encode proteins, which are predicted to have an effect on the contractile activity of myocardial cells, which in turn may have an effect on valvular performance and hemodynamics across the mitral valve. This may result in shear forces with impact on MMVD progression. [ABSTRACT FROM AUTHOR]

Aims: Alzheimer's disease (AD) is characterized by the accumulation of amyloid beta (Aβ) in the brain. The deposition of Aβ is believed to initiate a detrimental cascade, including cerebral hypometabolism, accelerated brain atrophy, and cognitive problems—ultimately resulting in AD. However, the timing and causality of the cascade resulting in AD are not yet fully established. Therefore, we examined whether early Aβ accumulation affects cerebral glucose metabolism, atrophy rate, and age-related cognitive decline before the onset of neurodegenerative disease. Methods: Participants from the Metropolit 1953 Danish Male Birth Cohort underwent brain positron emission tomography (PET) imaging using the radiotracers [11C]Pittsburgh Compound-B (PiB) (N = 70) and [18F]Fluorodeoxyglucose (FDG) (N = 76) to assess cerebral Aβ accumulation and glucose metabolism, respectively. The atrophy rate was calculated from anatomical magnetic resonance imaging (MRI) scans conducted presently and 10 years ago. Cognitive decline was examined from neurophysiological tests conducted presently and ten or 5 years ago. Results: Higher Aβ accumulation in AD-critical brain regions correlated with greater visual memory decline (p = 0.023). Aβ accumulation did not correlate with brain atrophy rates. Increased cerebral glucose metabolism in AD-susceptible regions correlated with worse verbal memory performance (p = 0.040). Conclusions: Aβ accumulation in known AD-related areas was associated with subtle cognitive deficits. The association was observed before hypometabolism or accelerated brain atrophy, suggesting that Aβ accumulation is involved early in age-related cognitive dysfunction. The association between hypermetabolism and worse memory performance may be due to early compensatory mechanisms adapting for malfunctioning neurons by increasing metabolism. [ABSTRACT FROM AUTHOR]

We document that in unstressed, undeformed, samples of pure iron containing silica inclusions precipitated by melt deoxidation, the iron matrix and a small fraction of the silica inclusions are locally separated at room temperature by a void. Thermal cycling, picnometry, dilatometry, and nano-holotomography experiments are combined to demonstrate that those voids originate from silica inclusion/iron matrix interfacial debonding during cooldown from elevated processing temperatures, resulting from the volumetric expansion that accompanies the austenite-to-ferrite phase transformation. [ABSTRACT FROM AUTHOR]

Background: We examined the impact of early (0–4 weeks after discharge) versus late (> 4–8 weeks after discharge) initiation of adjuvant chemotherapy on pancreatic adenocarcinoma survival. Methods: We used Danish population-based healthcare registries to emulate a hypothetical target trial using the clone-censor-weight approach. All eligible patients were cloned with one clone assigned to 'early initiation' and one clone assigned to 'late initiation'. Clones were censored when the assigned treatment was no longer compatible with the actual treatment. Informative censoring was addressed using inverse probability of censoring weighting. Results: We included 1491 patients in a hypothetical target trial, of whom 32.3% initiated chemotherapy within 0–4 weeks and 38.3% between > 4 and 8 weeks after discharge for pancreatic adenocarcinoma surgery; 206 (13.8%) initiated chemotherapy after > 8 weeks, and 232 (15.6%) did not initiate chemotherapy. Median overall survival was 30.4 and 29.9 months in late and early initiators, respectively. The absolute differences in OS, comparing late with early initiators, were 3.2% (95% confidence interval [CI] − 1.5%, 7.9%), − 0.7% (95% CI − 7.2%, 5.8%), and 3.2% (95% CI − 2.8%, 9.3%) at 1, 3, and 5 years, respectively. Late initiators had a higher increase in albumin levels as well as higher pretreatment albumin values. Conclusions: Postponement of adjuvant chemotherapy up to 8 weeks after discharge from pancreatic adenocarcinoma surgery is safe and may allow more patients to receive adjuvant therapy due to better recovery. [ABSTRACT FROM AUTHOR]

Objectives: Lung volume reduction surgery (LVRS) has proven an effective treatment for emphysema, by decreasing hyperinflation and improving lung function, activity level and reducing dyspnoea. However, postoperative air leak is an important complication, often leading to reoperation. Our aim was to analyse reoperations after LVRS and identify potential predictors. Methods: Consecutive single-centre unilateral VATS LVRS performed from 2017 to 2022 were included. Typically, 3–5 minor resections were made using vascular magazines without buttressing. Data were obtained from an institutional database and analysed. Multivariable logistic regression was used to identify predictors of reoperation. Number and location of injuries were registered. Results: In total, 191 patients were included, 25 were reoperated (13%). In 21 patients, the indication for reoperation was substantial air leak, 3 patients bleeding and 1 patient empyema. Length of stay (LOS) was 21 (11–33) vs. 5 days (3–11), respectively. Only 3 injuries were in the stapler line, 13 within < 2cm and 15 injuries were in another site. Multivariable logistic regression analysis showed that decreasing DLCO increased risk of reoperation, OR 1.1 (1.03, 1.18, P = 0.005). Resections in only one lobe, compared to resections in multiple lobes, were also a risk factor OR 3.10 (1.17, 9.32, P = 0.03). Patients undergoing reoperation had significantly increased 30-day mortality, OR 5.52 (1.03, 26.69, P = 0.02). Conclusions: Our incidence of reoperation after LVRS was 13% leading to prolonged LOS and increased 30-day mortality. Low DLCO and resections in a single lobe were significant predictors of reoperation. The air leak was usually not localized in the stapler line. Key question: What characterizes reoperations after lung volume reduction surgeries? Key findings: Lung injuries were predominantly located away from the original surgical site. Take home message: Lung injuries remote from the stapler line is frequent during reoperation after lung volume reduction surgery. Location of lung injuries found during reoperation after lung volume reduction surgery [ABSTRACT FROM AUTHOR]

Neutrophils play a significant role in sustaining chronic inflammation in Inflammatory Bowel Disease. The intestinal basement membrane acts as a barrier for immunological homeostasis, where the α3 and α4 chains of type IV collagen are expressed on the mucosal surface. We wanted to develop a biomarker reflecting early tissue injury, providing an opportunity for intervention. Two competitive enzyme-linked immunosorbent assays (ELISAs) quantifying human neutrophil elastase (HNE) degraded neo-epitopes of COL4A3 and COL4A4 were developed and investigated in two observational cohorts (n = 161, n = 100). A biomarker of MMP-mediated degradation of COL4A1 (C4M) was used for comparison. In Cohort 1, patients with mild endoscopic ulcerative colitis showed elevated levels of C4A3-HNE compared to those with severe disease. C4M had a strong positive correlation with disease activity. C4A3-HNE/C4M provided superior discrimination between mild and severe endoscopic disease and negatively correlated to disease activity. In Cohort 2, C4A4-HNE and C4A4-HNE/C4M showed similar trends. C4A3-HNE and C4A4-HNE possibly reflect early intestinal tissue injury. Combining the markers with a biomarker of another α-chain of the same collagen provides information on two distinct stages of mucosal damage. These biomarkers may be used to monitor disease flare-up in patients in remission, reducing the need for frequent endoscopic procedures. [ABSTRACT FROM AUTHOR]

Major migration events in Holocene Eurasia have been characterized genetically at broad regional scales1–4. However, insights into the population dynamics in the contact zones are hampered by a lack of ancient genomic data sampled at high spatiotemporal resolution5–7. Here, to address this, we analysed shotgun-sequenced genomes from 100 skeletons spanning 7,300 years of the Mesolithic period, Neolithic period and Early Bronze Age in Denmark and integrated these with proxies for diet (13C and 15N content), mobility (87Sr/86Sr ratio) and vegetation cover (pollen). We observe that Danish Mesolithic individuals of the Maglemose, Kongemose and Ertebølle cultures form a distinct genetic cluster related to other Western European hunter-gatherers. Despite shifts in material culture they displayed genetic homogeneity from around 10,500 to 5,900 calibrated years before present, when Neolithic farmers with Anatolian-derived ancestry arrived. Although the Neolithic transition was delayed by more than a millennium relative to Central Europe, it was very abrupt and resulted in a population turnover with limited genetic contribution from local hunter-gatherers. The succeeding Neolithic population, associated with the Funnel Beaker culture, persisted for only about 1,000 years before immigrants with eastern Steppe-derived ancestry arrived. This second and equally rapid population replacement gave rise to the Single Grave culture with an ancestry profile more similar to present-day Danes. In our multiproxy dataset, these major demographic events are manifested as parallel shifts in genotype, phenotype, diet and land use.Integrated data, including 100 human genomes from the Mesolithic, Neolithic and Early Bronze Age periods show that two major population turnovers occurred over just 1,000 years in Neolithic Denmark, resulting in dramatic changes in the genes, diet and physical appearance of the local people, as well as the landscape in which they lived. [ABSTRACT FROM AUTHOR]

The health status of cold-water coral reefs on the West African coast was investigated with the main objective of obtaining knowledge of the adaptive capacity of Lophelia pertusa to environmental stressors. Three coral sites were studied, in Northern Morocco, in the Morocco/Mauritania region (both in 2020) and, in the Ghana and Ivory coast region (visited in 2012, 2017, and 2019). Area cover of live colonies, explored through underwater videos, was used as an indicator of reef health and compared with the environmental variables: reef position, depth, water mass, temperature, dissolved oxygen concentration (DO), carbonate chemistry (pH, aragonite saturation (ΩAr), macronutrients and particles (visual). For a broader picture of the adaptations presented by Lophelia our results were compared with reefs in contrasting environments. Off Ghana and Mauritania healthy reefs (i.e., having areas with more than 20 % cover of live colonies) were found to reside at DO concentrations between 1.1 and 1.6 ml L−1, in corrosive waters (pH 7.7 and ΩAr 1.0) with high nutrient concentrations. By contrast, the reefs off the North of Morocco, sitting in well-oxygenated waters with oversaturated ΩAr, had no or few live colonies. Our findings together with data from other studies show that Lophelia has a wide tolerance to hypoxia and acidification, and that in relation to climate change increased temperature and silting could pose more serious threats. These findings highlight the importance of continued studies of Lophelia reefs in contrasting environmental conditions to better understand their adaptation potential to climate change-related stressors. [ABSTRACT FROM AUTHOR]

Purpose: We assessed long-term outcomes in acutely admitted adult patients with delirium treated in intensive care unit (ICU) with haloperidol versus placebo. Methods: We conducted pre-planned analyses of 1-year outcomes in the Agents Intervening against Delirium in the ICU (AID-ICU) trial, including mortality and health-related quality of life (HRQoL) assessed by Euroqol (EQ) 5-dimension 5-level questionnaire (EQ-5D-5L) index values and EQ visual analogue scale (EQ VAS) (deceased patients were assigned the numeric value zero). Outcomes were analysed using logistic and linear regressions with bootstrapping and G-computation, all with adjustment for the stratification variables (site and delirium motor subtype) and multiple imputations for missing HRQoL values. Results: At 1-year follow-up, we obtained vital status for 96.2% and HRQoL data for 83.3% of the 1000 randomised patients. One-year mortality was 224/501 (44.7%) in the haloperidol group versus 251/486 (51.6%) in the placebo group, with an adjusted absolute risk difference of − 6.4%-points (95% confidence interval [CI] − 12.8%-points to − 0.2%-points; P = 0.045). These results were largely consistent across the secondary analyses. For HRQoL, the adjusted mean differences were 0.04 (95% CI − 0.03 to 0.11; P = 0.091) for EQ-5D-5L-5L index values, and 3.3 (95% CI − 9.3 to 17.5; P = 0.142) for EQ VAS. Conclusions: In acutely admitted adult ICU patients with delirium, haloperidol treatment reduced mortality at 1-year follow-up, but did not statistically significantly improve HRQoL. [ABSTRACT FROM AUTHOR]

Adverse outcome pathways (AOPs) provide evidence for demonstrating and assessing causality between measurable toxicological mechanisms and human or environmental adverse effects. AOPs have gained increasing attention over the past decade and are believed to provide the necessary steppingstone for more effective risk assessment of chemicals and materials and moving beyond the need for animal testing. However, as with all types of data and knowledge today, AOPs need to be reusable by machines, i.e., machine-actionable, in order to reach their full impact potential. Machine-actionability is supported by the FAIR principles, which guide findability, accessibility, interoperability, and reusability of data and knowledge. Here, we describe why AOPs need to be FAIR and touch on aspects such as the improved visibility and the increased trust that FAIRification of AOPs provides. Plain language summary New approach methodologies (NAMs) can detect biological phenomena that occur before they add up to serious problems like cancer, infertility, death, and others. NAMs detect key events (KE) along well-proven and agreed adverse outcome pathways (AOP). If a substance tests positive in a NAM for an upstream KE, this signals an early warning that actual adversity might follow. However, what if the knowledge about these AOPs is a well-kept secret? And what if decision-makers find AOPs too exotic to apply in risk assessment? This is where FAIR comes in! FAIR stands for making information findable, accessible, interoperable and re-useable. It aims to increase availability, usefulness, and trustworthiness of data. Here, we show that by interpreting the FAIR principles beyond a purely technical level, AOPs can ring in a new era of 3Rs applicability by increasing their visibility and making their creation process more transparent and reproducible. [ABSTRACT FROM AUTHOR]

Class-switching to IgG2a/c in mice is a hallmark response to intracellular pathogens. T cells can promote class-switching and the predominant pathway for induction of IgG2a/c antibody responses has been suggested to be via stimulation from Th1 cells. We previously formulated CAF®01 (cationic liposomes containing dimethyldioctadecylammonium bromide (DDA) and Trehalose-6,6-dibehenate (TDB)) with the lipidated TLR7/8 agonist 3M-052 (DDA/TDB/3M-052), which promoted robust Th1 immunity in newborn mice. When testing this adjuvant in adult mice using the recombinant Chlamydia trachomatis (C.t.) vaccine antigen CTH522, it similarly enhanced IgG2a/c responses compared to DDA/TDB, but surprisingly reduced the magnitude of the IFN-γ+Th1 response in a TLR7 agonist dose-dependent manner. Single-cell RNA-sequencing revealed that DDA/TDB/3M-052 liposomes initiated early transcription of class-switch regulating genes directly in pre-germinal center B cells. Mixed bone marrow chimeras further demonstrated that this adjuvant did not require Th1 cells for IgG2a/c switching, but rather facilitated TLR7-dependent T-bet programming directly in B cells. This study underlines that adjuvant-directed IgG2a/c class-switching in vivo can occur in the absence of T-cell help, via direct activation of TLR7 on B cells and positions DDA/TDB/3M-052 as a powerful adjuvant capable of eliciting type I-like immunity in B cells without strong induction of Th1 responses. [ABSTRACT FROM AUTHOR]

The mechanisms leading to changes in mesoscale chromatin organization during cellular aging are unknown. Here, we used transcriptional activator-like effectors, RNA-seq and superresolution analysis to determine the effects of genotoxic stress on oocyte chromatin structure. Major satellites are organized into tightly packed globular structures that coalesce into chromocenters and dynamically associate with the nucleolus. Acute irradiation significantly enhanced chromocenter mobility in transcriptionally inactive oocytes. In transcriptionally active oocytes, irradiation induced a striking unfolding of satellite chromatin fibers and enhanced the expression of transcripts required for protection from oxidative stress (Fermt1, Smg1), recovery from DNA damage (Tlk2, Rad54l) and regulation of heterochromatin assembly (Zfp296, Ski-oncogene). Non-irradiated, senescent oocytes exhibit not only high chromocenter mobility and satellite distension but also a high frequency of extra chromosomal satellite DNA. Notably, analysis of biological aging using an oocyte-specific RNA clock revealed cellular communication, posttranslational protein modifications, chromatin and histone dynamics as the top cellular processes that are dysregulated in both senescent and irradiated oocytes. Our results indicate that unfolding of heterochromatin fibers following acute genotoxic stress or cellular aging induced the formation of distended satellites and that abnormal chromatin structure together with increased chromocenter mobility leads to chromosome instability in senescent oocytes. Super-resolution imaging and RNA clock analyses show that exposure of mouse oocytes to ionizing radiation compromises chromatin organization and dramatically accelerates biological aging. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the spatial distribution of micropollutants in wastewater related to catchment area, and their environmental risks and fate. About 24-h flow proportional effluent (n = 26) wastewater samples were collected from eight WWTPs across Denmark. From five of these WWTPs corresponding influent samples (n = 20) were collected. Samples were enriched by multi-layer solid phase and analysed by liquid chromatography–high-resolution mass spectrometry and comprehensive two-dimensional gas chromatography with high-resolution mass spectrometry detection. We detected and quantified 79 micropollutants from a list of 291 micropollutants in at least one influent or effluent wastewater sample. From this we found that 54 micropollutants decreased in concentrations during wastewater treatment, while O-desmethylvenlafaxine, carbamazepine, amitriptyline, benzothiazole, terbutryn, and citalopram increased in concentrations through the WWTP. The toxicity of effluent wastewater samples was assessed by EC50 using Raphidocelis subcapitata (R. subcapitata) and LC50 using the crustacean Daphnia magna (D. Magna), for which six micropollutants were detected above the predicted no-effect concentration. Our study demonstrates that catchment area influences the micropollutant composition of wastewater. Out of 19 pharmaceuticals, the measured concentration in influent wastewater was predicted within a factor of 10 from sale numbers and human excretion, which demonstrates the strong influence of catchment area on micropollutant composition. [ABSTRACT FROM AUTHOR]

Larval cod require live prey as food, and prey concentration (PC) and feeding frequency (FF) often affect their growth and survival. Apart from this, water exchange rates/water flow (WER/WF) and water current (WC) also affect the prey resident time in the tank and larval/early juvenile behaviour, respectively. High water current is also known to induce stress in finfish larvae, and this stress response is believed to be dependent on larval developmental stage. Thus, we conducted a study to evaluate three larval rearing protocols varying in prey concentration, feeding frequency, and water current/exchange rate. Three protocols were used: low prey concentration (PC), low feeding frequency (FF), and low water flow (protocol 1); medium PC, medium FF, and medium WF (protocol 2); and high PC, high FF, and high WF (protocol 3) (see Table 1). Larvae were sampled periodically for growth and cortisol measurements. Cortisol extraction and radioimmunoassay (RIA) were conducted using methods previously validated for cod larvae. Larvae reared using protocols 2 and 3 were significantly bigger and heavier than larvae reared using protocol 1. Rearing protocol had a significant effect on the cortisol level in larval cod. Larvae showed a developmental stage-dependent stress response. Protocol 2 had significantly higher survival than protocols 1 and 3. Our results indicate that an intermediate PC, FF, and WF (protocol 2) is suitable during cod larval rearing. [ABSTRACT FROM AUTHOR]

Background: Feedback is important for surgical trainees but it can be biased and time-consuming. We examined crowd-sourced assessment as an alternative to experienced surgeons' assessment of robot-assisted radical prostatectomy (RARP). Methods: We used video recordings (n = 45) of three RARP modules on the RobotiX, Simbionix simulator from a previous study in a blinded comparative assessment study. A group of crowd workers (CWs) and two experienced RARP surgeons (ESs) evaluated all videos with the modified Global Evaluative Assessment of Robotic Surgery (mGEARS). Results: One hundred forty-nine CWs performed 1490 video ratings. Internal consistency reliability was high (0.94). Inter-rater reliability and test–retest reliability were low for CWs (0.29 and 0.39) and moderate for ESs (0.61 and 0.68). In an Analysis of Variance (ANOVA) test, CWs could not discriminate between the skill level of the surgeons (p = 0.03–0.89), whereas ES could (p = 0.034). Conclusion: We found very low agreement between the assessments of CWs and ESs when they assessed robot-assisted radical prostatectomies. As opposed to ESs, CWs could not discriminate between surgical experience using the mGEARS ratings or when asked if they wanted the surgeons to perform their robotic surgery. [ABSTRACT FROM AUTHOR]

Thyroid cancer is the most common endocrine malignancy, affecting nearly 600,000 new patients worldwide. Treatment with the BRAF inhibitor sorafenib partially prolongs progression-free survival in thyroid cancer patients, but fails to improve overall survival. This study examines enhancing sorafenib efficacy by combination therapy with the novel HSP90 inhibitor onalespib. In vitro efficacy of sorafenib and onalespib monotherapy as well as in combination was assessed in papillary (PTC) and anaplastic (ATC) thyroid cancer cells using cell viability and colony formation assays. Migration potential was studied in wound healing assays. The in vivo efficacy of sorafenib and onalespib therapy was evaluated in mice bearing BHT-101 xenografts. Sorafenib in combination with onalespib significantly inhibited PTC and ATC cell proliferation, decreased metabolic activity and cancer cell migration. In addition, the drug combination approach significantly inhibited tumor growth in the xenograft model and prolonged the median survival. Our results suggest that combination therapy with sorafenib and onalespib could be used as a new therapeutic approach in the treatment of thyroid cancer, significantly improving the results obtained with sorafenib as monotherapy. This approach has the potential to reduce treatment adaptation while at the same time providing therapeutic anti-cancer benefits such as reducing tumor growth and metastatic potential. [ABSTRACT FROM AUTHOR]

Objectives : Machine learning (ML) for medical imaging is emerging for several organs and image modalities. Our objectives were to provide clinicians with an overview of this field by answering the following questions: (1) How is ML applied in liver computed tomography (CT) imaging? (2) How well do ML systems perform in liver CT imaging? (3) What are the clinical applications of ML in liver CT imaging? Methods: A systematic review was carried out according to the guidelines from the PRISMA-P statement. The search string focused on studies containing content relating to artificial intelligence, liver, and computed tomography. Results: One hundred ninety-one studies were included in the study. ML was applied to CT liver imaging by image analysis without clinicians' intervention in majority of studies while in newer studies the fusion of ML method with clinical intervention have been identified. Several were documented to perform very accurately on reliable but small data. Most models identified were deep learning-based, mainly using convolutional neural networks. Potentially many clinical applications of ML to CT liver imaging have been identified through our review including liver and its lesion segmentation and classification, segmentation of vascular structure inside the liver, fibrosis and cirrhosis staging, metastasis prediction, and evaluation of chemotherapy. Conclusion: Several studies attempted to provide transparent result of the model. To make the model convenient for a clinical application, prospective clinical validation studies are in urgent call. Computer scientists and engineers should seek to cooperate with health professionals to ensure this. Key Points: • ML shows great potential for CT liver image tasks such as pixel-wise segmentation and classification of liver and liver lesions, fibrosis staging, metastasis prediction, and retrieval of relevant liver lesions from similar cases of other patients. • Despite presenting the result is not standardized, many studies have attempted to provide transparent results to interpret the machine learning method performance in the literature. • Prospective studies are in urgent call for clinical validation of ML method, preferably carried out by cooperation between clinicians and computer scientists. [ABSTRACT FROM AUTHOR]

Molecular radiotherapy combines the advantages of systemic administration of highly specific antibodies or peptides and the localized potency of ionizing radiation. A potential target for molecular radiotherapy is the cell surface antigen CD44v6, which is overexpressed in numerous cancers, with limited expression in normal tissues. The aim of the present study was to generate and characterize a panel of human anti-CD44v6 antibodies and identify a suitable candidate for future use in molecular radiotherapy of CD44v6-expressing cancers. Binders were first isolated from large synthetic phage display libraries containing human scFv and Fab antibody fragments. The antibodies were extensively analyzed through in vitro investigations of binding kinetics, affinity, off-target binding, and cell binding. Lead candidates were further subjected to in vivo biodistribution studies in mice bearing anaplastic thyroid cancer xenografts that express high levels of CD44v6. Additionally, antigen-dependent tumor uptake of the lead candidate was verified in additional xenograft models with varying levels of target expression. Interestingly, although only small differences were observed among the top antibody candidates in vitro, significant differences in tumor uptake and retention were uncovered in in vivo experiments. A high-affinity anti-CD44v6 lead drug candidate was identified, mAb UU-40, which exhibited favorable target binding properties and in vivo distribution. In conclusion, a panel of human anti-CD44v6 antibodies was successfully generated and characterized in this study. Through comprehensive evaluation, mAb UU-40 was identified as a promising lead candidate for future molecular radiotherapy of CD44v6-expressing cancers due to its high affinity, excellent target binding properties, and desirable in vivo distribution characteristics. [ABSTRACT FROM AUTHOR]

Proportional hazards models have been proposed to analyse time-to-event phenotypes in genome-wide association studies (GWAS). However, little is known about the ability of proportional hazards models to identify genetic associations under different generative models and when ascertainment is present. Here we propose the age-dependent liability threshold (ADuLT) model as an alternative to a Cox regression based GWAS, here represented by SPACox. We compare ADuLT, SPACox, and standard case-control GWAS in simulations under two generative models and with varying degrees of ascertainment as well as in the iPSYCH cohort. We find Cox regression GWAS to be underpowered when cases are strongly ascertained (cases are oversampled by a factor 5), regardless of the generative model used. ADuLT is robust to ascertainment in all simulated scenarios. Then, we analyse four psychiatric disorders in iPSYCH, ADHD, Autism, Depression, and Schizophrenia, with a strong case-ascertainment. Across these psychiatric disorders, ADuLT identifies 20 independent genome-wide significant associations, case-control GWAS finds 17, and SPACox finds 8, which is consistent with simulation results. As more genetic data are being linked to electronic health records, robust GWAS methods that can make use of age-of-onset information will help increase power in analyses for common health outcomes. Robust genome-wide association study (GWAS) methods that can utilise time-to-event information such as age-of-onset will help increase power in analyses for common health outcomes. Here, the authors propose a computationally efficient time-to-event model for GWAS. [ABSTRACT FROM AUTHOR]

Purpose: The aim of the study was to perform a systematic review and best knowledge synthesis of the present literature concerning biomechanical risk factors for developing first-time and recurrent patella dislocation. Methods: The study was performed as a systematic review following PRISMA guidelines. PubMed and EMBASE were systematically searched. Studies investigating participants with risk factors for first-time as well as recurrent patella dislocation were included. The records were screened, and data extracted independently by two researchers supervised by a third independent assessor. The study was registered in PROSPERO. Results: A total of 6233 records were screened, and 50 studies met the inclusion criteria. The biomechanical risk factors: trochlear dysplasia, increased tibial tuberosity–trochlear groove distance (TT-TG), and patella alta were found to be statistically significantly associated with patella dislocation in several publications and were thus recognized as risk factors for patella dislocation. The soft-tissue stabilizers: longer and thinner MPFL ligament, increased number of type 2C and decreased number of type 1 muscle fibers, and joint laxity were found to be statistically significantly associated with patella dislocation in a few publications, but due to limited evidence, no conclusion was made on this matter. Conclusion: There is strong evidence in the literature that abnormalities of bony stabilizers, trochlear dysplasia, increased TT–TG distance, and patella alta are risk factors for patella dislocation. There is less evidence that soft-tissue stabilizers are risk factors. The study emphasizes the importance of a thorough investigation of bony stabilizers in clinical decision-making. Level of evidence: Level IV. [ABSTRACT FROM AUTHOR]

Purpose: The aim of our study was to perform a systematic review and best knowledge synthesis of the present literature concerning the familial association and epidemiological factors as risk factors for developing first-time and recurrent patella dislocation. Methods: The study was conducted according to the PRISMA guidelines and registered in PROSPERO. EMBASE and PubMed were systematically searched on the 5th of May 2022. Studies investigating participants with genetic and epidemiological risk factors for the first time as well as recurrent patella dislocation were included. The records were screened, and data were extracted independently by two researchers supervised by a third independent assessor. Results: A total of 6,649 records were screened, and 67 studies were included. Familial association was described as a risk factor for patella dislocation in 17 studies. One study found that participants with a family history of patella dislocation had a 3.7 higher risk for patella dislocation in the contralateral asymptomatic knee, and another study found a family history of PD in 9% of 74 participants. Eleven studies found an accumulation of patella dislocation across generations in specific families. Additionally, a range of genetic syndromes was associated with patella dislocation. Young age is a well-investigated risk factor for patella dislocation, but the results are inconsistent. Only five and eight studies investigated skeletal immaturity and gender as risk factors for patella dislocation, respectively. Conclusion: There may be a familial association with patella dislocation, but further investigation is necessary to determine the strength and etiology of the association. There is weak evidence that epidemiological risk factors, such as age, skeletal immaturity, gender, and BMI are risk factors for patella dislocation. Level of evidence: IV. [ABSTRACT FROM AUTHOR]

The upper limit for partial hepatectomy (PH) in rats is 90%, which is associated with an increased risk of post-hepatectomy liver failure (PHLF), correlating with high mortality. Sixty-eight rats were randomized to 90% PH, sham operation, or no surgery. Further block randomization was performed to determine the time of euthanasia, whether 12, 24, or 48 h after surgery. A general distress score (GDS) was calculated to distinguish between rats with reversible (GDS < 10) and irreversible PHLF (GDS ≥ 10). At euthanasia, the liver remnant and blood were collected. Liver-specific biochemistry and regeneration ratio were measured. Hepatocyte proliferation and volume were estimated using stereological methods. All rats subjected to 90% experienced biochemical PHLF. The biochemical and morphological liver responses did not differ between the groups until 48 h after surgery. At 48 h, liver regeneration and function were significantly improved in survivors. The peak mean regeneration ratio was 15% for rats with irreversible PHLF compared to 26% for rats with reversible PHLF. The 90% PH rat model was associated with PHLF and high mortality. Irreversible PHLF was characterized by impaired liver regeneration capacity and an insufficient ability to metabolize ammonia. [ABSTRACT FROM AUTHOR]

The predictive performance of polygenic scores (PGS) is largely dependent on the number of samples available to train the PGS. Increasing the sample size for a specific phenotype is expensive and takes time, but this sample size can be effectively increased by using genetically correlated phenotypes. We propose a framework to generate multi-PGS from thousands of publicly available genome-wide association studies (GWAS) with no need to individually select the most relevant ones. In this study, the multi-PGS framework increases prediction accuracy over single PGS for all included psychiatric disorders and other available outcomes, with prediction R2 increases of up to 9-fold for attention-deficit/hyperactivity disorder compared to a single PGS. We also generate multi-PGS for phenotypes without an existing GWAS and for case-case predictions. We benchmark the multi-PGS framework against other methods and highlight its potential application to new emerging biobanks. Polygenic scores (PGS) have high potential for clinical use but are currently underpowered for many applications. Here, the authors develop an approach that leverages an agnostic library of hundreds of PGS to increase prediction of complex diseases and other traits. This multi-PGS framework is ideal for emerging biobank data. [ABSTRACT FROM AUTHOR]

Volumetric muscle loss (VML) results in permanent functional deficits and remains a substantial regenerative medicine challenge. A coordinated immune response is crucial for timely myofiber regeneration, however the immune response following VML has yet to be fully characterized. Here, we leveraged dimensionality reduction and pseudo-time analysis techniques to elucidate the cellular players underlying a functional or pathological outcome as a result of subcritical injury or critical VML in the murine quadriceps, respectively. We found that critical VML resulted in a sustained presence of M2-like and CD206hiLy6Chi 'hybrid' macrophages whereas subcritical defects resolved these populations. Notably, the retained M2-like macrophages from critical VML injuries presented with aberrant cytokine production which may contribute to fibrogenesis, as indicated by their co-localization with fibroadipogenic progenitors (FAPs) in areas of collagen deposition within the defect. Furthermore, several T cell subpopulations were significantly elevated in critical VML compared to subcritical injuries. These results demonstrate a dysregulated immune response in critical VML that is unable to fully resolve the chronic inflammatory state and transition to a pro-regenerative microenvironment within the first week after injury. These data provide important insights into potential therapeutic strategies which could reduce the immune cell burden and pro-fibrotic signaling characteristic of VML. The immune cell dynamics that influence the pathological phenotype of a critical volumetric muscle loss (VML) injury that cannot regenerate versus a subcritical injury capable of regeneration are characterized using quadriceps VML mouse models. [ABSTRACT FROM AUTHOR]

We report here the discovery and structure-guided optimization of a novel series of AKT kinase inhibitors. Based on docking studies for the predicted active bound-conformation of 2, a potent series of N-substituted-5-(isoquinolin-6-yl)-1,3,4-thiadiazol-2-amines was developed. Compounds in the series achieve AKT pathway inhibition in cancer cells, as measured by inhibition of pathway proteins pGSK and pFKHR. Compound 12 was further evaluated in a single dose PK/PD in vivo study in tumor-bearing mice and demonstrated inhibition of phosphorylation of the direct substrate GSK and pathway biomarker S6. [ABSTRACT FROM AUTHOR]

Purpose: The purpose was to investigate the diagnostic performance of bimodal optical and radio-guided sentinel node biopsy (SNB) for oral squamous cell carcinoma (OSCC) sub-sites in the anterior oral cavity. Methods: Prospective study of 50 consecutive patients with cN0 OSCC scheduled for SNB was injected with the tracer complex Tc99m:ICG:Nacocoll. A near-infrared camera was applied for optical SN detection. Endpoints were modality for intraoperative SN detection and false omission rate at follow-up. Results: In all patients, a SN could be detected. In 12/50 (24%) of cases, the SPECT/CT showed no focus in level 1, but intraoperatively a SN in level 1 was optically detected. In 22/50 cases (44%), an additional SN was identified only due to the optical imaging. At follow-up, the false omission rate was 0%. Conclusion: Optical imaging appears to be an effective tool to allow real-time SN identification comprising level 1 unaffected by possible interference of radiation site from the injection. [ABSTRACT FROM AUTHOR]

Background: Patients with gastric adenocarcinoma (GAC) are at high risk of peritoneal recurrence despite perioperative chemotherapy and radical resection. This study evaluated feasibility and safety of laparoscopic D2 gastrectomy in combination with pressurized intraperitoneal aerosol chemotherapy (PIPAC). Methods: This was a prospective, controlled bi-institutional study in patients with GAC at high risk of recurrence treated with PIPAC with cisplatin and doxorubicin (PIPAC C/D) after laparoscopic D2 gastrectomy. High risk was defined as a poorly cohesive subtype with predominance of signet-ring cells, clinical stage ≥ T3 and/or ≥ N2, or positive peritoneal cytology. Peritoneal lavage fluid was collected before and after resection. Cisplatin (10.5 mg/m2) and doxorubicin (2.1 mg/m2) were aerosolized after anastomosis (flow 0.5–0.8 ml/s, maximum pressure 300 PSI). Treatment was feasible and safe if ≤ 20% had Dindo–Clavien ≥ 3b surgical complications or CTCAE ≥ 4 medical adverse events within 30 days. Secondary outcomes were length of stay (LOS), peritoneal lavage cytology, and completion of postoperative systemic chemotherapy. Results: Twenty-one patients were treated with a D2 gastrectomy and PIPAC C/D. The median age was 61 years (range 24–76), there were eleven female patients, and 20 patients had preoperative chemotherapy. There was no mortality. Two patients had grade 3b complications that were potentially related to PIPAC C/D (one anastomotic leakage, and one late duodenal blow-out). One patient had severe neutropenia, and nine patients had moderate pain. The LOS was 6 days (4–26). One patient had positive peritoneal lavage cytology before resection, and none were positive after. Fifteen patients had postoperative chemotherapy. Conclusions: Laparoscopic D2 gastrectomy in combination with PIPAC C/D is feasible and safe. [ABSTRACT FROM AUTHOR]

Aims: Gamma oscillations (≈25–100 Hz) are believed to play an essential role in cognition, and aberrant gamma oscillations occur in brain aging and neurodegeneration. This study examined age-related changes in visually evoked gamma oscillations at two different time points 5 years apart and tested the hypothesis that the power of gamma oscillations correlated to cognitive skills. Methods: The cohort consists of elderly males belonging to the Metropolit 1953 Danish Male Birth Cohort (first visit, N=124; second visit, N=88) over a 5-year period from 63 to 68 years of age. Cognitive functions were assessed using a neuropsychological test battery measuring global cognition, intelligence, memory, and processing speed. The power of steady-state visual evoked potentials (SSVEP) was measured at 8 Hz (alpha) and 36 Hz (gamma) frequencies using EEG scalp electrodes. Results: Over the 5-year period cognitive performance remained relatively stable while the power of visually evoked gamma oscillations shifted from posterior to anterior brain regions with increasing age. A higher-than-average cognitive score was correlated with higher gamma power in parieto-occipital areas and lower in frontocentral areas, i.e., preserved distribution of the evoked activity. Conclusions: Our data reveal that the distribution of visually evoked gamma activity becomes distributed with age. Preserved posterior-occipital gamma power in participants with a high level of cognitive performance is consistent with a close association between the ability to produce gamma oscillations and cognition. The data may contribute to our understanding of the mechanisms that link evoked gamma activity and cognition in the aging brain. [ABSTRACT FROM AUTHOR]

The only licensed tuberculosis (TB) vaccine, Bacillus Calmette Guerin (BCG), fails to reliably protect adolescents and adults from pulmonary TB, resulting in ~1.6 million deaths annually. Protein subunit vaccines have shown promise against TB in clinical studies. Unfortunately, most subunit vaccines require multiple administrations, which increases the risk of loss to follow-up and necessitates more complex and costly logistics. Given the well-documented adjuvant effect of BCG, we hypothesized that BCG co-administration could compensate for a reduced number of subunit vaccinations. To explore this, we developed an expression-optimized version of our H107 vaccine candidate (H107e), which does not cross-react with BCG. In the CAF®01 adjuvant, a single dose of H107e induced inferior protection compared to three H107e/CAF®01 administrations. However, co-administering a single dose of H107e/CAF®01 with BCG significantly improved protection, which was equal to BCG co-administered with three H107e/CAF®01 doses. Importantly, combining BCG with a single H107e/CAF®01 dose also increased protection in previously BCG-primed animals. Overall, a single dose of H107e/CAF®01 with BCG induced long-lived immunity and triggered BCG-specific Th17 responses. These data support co-administration of BCG and subunit vaccines in both BCG naïve and BCG-primed individuals as an improved TB vaccine strategy with reduced number of vaccination visits. [ABSTRACT FROM AUTHOR]

Background: Pressurized Intraperitoneal Aerosol chemotherapy (PIPAC) is a local treatment for peritoneal metastasis (PM). Prospective data are scarce and evaluation of treatment response remains difficult. This study evaluated the use of the Peritoneal Regression Grading score (PRGS) and its prognostic value. Patients and Methods: This was a prospective, controlled phase II trial in patients with PM from gastrointestinal, gynaecological, hepatopancreatobiliary, primary peritoneal, or unknown primary cancer. Patients in performance status 0–1, with a non-obstructed gastrointestinal tract, and a maximum of one extraperitoneal metastasis were eligible. Colorectal or appendiceal PM had PIPAC with oxaliplatin, other primaries had PIPAC with cisplatin and doxorubicin. Biopsies were taken at each PIPAC and evaluated using the PRGS. Quality-of-life questionnaires were reported at baseline and after three PIPACs. Results: One hundred ten patients were treated with 336 PIPACs (median 3, range 1–12). One hundred patients had prior palliative chemotherapy and 45 patients received bidirectional treatment. Complete or major histological response to treatment (PRGS 1–2) was observed in 38 patients (61%) who had three PIPACs, which was the only independent prognostic factor in a multivariate analysis. The median overall survival (mOS) from PIPAC 1 was 10 months, while patients with PM from gastric, colorectal, and pancreatic cancer had a mOS of 7.4, 16.7, and 8.2 months, respectively. Global health scores were significantly reduced, but patients were less fatigued, nauseated, constipated, and had better appetite after three PIPACs. Conclusions: PIPAC with oxaliplatin or cisplatin and doxorubicin was able to induce a major or complete histological response during three PIPACs, which may provide significant prognostic information, both at baseline and after treatment. [ABSTRACT FROM AUTHOR]

Early intervention and short-duration treatments with antibiotics in premature infants are reported to reduce the incidence of necrotizing enterocolitis (NEC), a terrible disease with severe inflammation and impaired intestinal barrier properties. Yet, it is unclear how antibiotics exposure, as well as route of administration used for dosing, can minimize the risk of NEC. With this study, we aimed to investigate if and how administration of antibiotics may affect the barrier properties of intestinal mucosa and mucus. We compared how parenteral (PAR) and a combination of enteral and parenteral (ENT+PAR) ampicillin and gentamicin given to preterm born piglets within 48 h after birth affected both barrier and physical properties of ex vivo small intestinal mucosa and mucus. Permeation of the markers mannitol, metoprolol, and fluorescein-isothiocyanate dextran of 4 kDa (FD4) and 70 kDa (FD70) through the mucosa and mucus was evaluated. For all markers, permeation through the mucosa and mucus collected from PAR piglets tended to be reduced when compared to that observed using untreated piglets. In contrast, permeation through the mucosa and mucus collected from ENT+PAR piglets tended to be similar to that observed for untreated piglets. Additionally, rheological measurements on the mucus from PAR piglets and ENT+PAR piglets displayed a decreased Gʹ and Gʹ/Gʺ ratio and decreased viscosity at 0.4 s−1 as well as lower stress stability compared to the mucus from untreated piglets. [ABSTRACT FROM AUTHOR]

Biologics, such as antibodies and enzymes, are crucial in research, biotechnology, diagnostics, and therapeutics. Often, biologics with suitable functionality are discovered, but their development is impeded by developability issues. Stability and solubility are key biophysical traits underpinning developability potential, as they determine aggregation, correlate with production yield and poly-specificity, and are essential to access parenteral and oral delivery. While advances for the optimisation of individual traits have been made, the co-optimization of multiple traits remains highly problematic and time-consuming, as mutations that improve one property often negatively impact others. In this work, we introduce a fully automated computational strategy for the simultaneous optimisation of conformational stability and solubility, which we experimentally validate on six antibodies, including two approved therapeutics. Our results on 42 designs demonstrate that the computational procedure is highly effective at improving developability potential, while not affecting antigen-binding. We make the method available as a webserver at www-cohsoftware.ch.cam.ac.uk. Antibodies find key applications in research, diagnostics, and therapeutics, but their development can be impeded by poor stability or solubility. Here the authors developed a computational strategy that enables antibody optimisation, without affecting functionality. [ABSTRACT FROM AUTHOR]