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It has been well documented that the content of the ECM can either induce or inhibit bone formation. For example, the size of bone matrix particles and the structure of collagen fibrils were found to affect the lineage commitment of MSCs . Meanwhile, the shape, crystallinity, surface charge, solubility, and mechanical properties of HA nanoparticles could also influence osteoblastic activities and the osteogenic potential of MSCs . It was theorized that MSCs were recruited to osteoclast-mediated bone resorption pits and contacted with the debris after resorption. HA nanoparticles were postulated to be one of the major components of the debris. Differentially shaped HA nanoparticles were suggested to provide dissimilar geometric cues and determine the fate of osteoblasts and MSCs. This was supported by findings suggesting that cigarette-like HA nanoparticles in OA bones could promote cytoskeletal changes of human MSCs and enhance their osteogenic potentials . As there were no significant differences between the HA nanoparticles in healthy and OA bone except for their shape, it is possible that the physical property of HA, rather than the chemical property, is deterministic for controlling the fate of MSCs in bone remodeling. Hence, osteocyte dysfunction may also indirectly promote osteogenic differentiation of MSCs through inducing aberrant mineralization by osteoblasts.
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The hybrid POM compounds, namely Ni-PYI, were isolated by the reaction of [BW12O40]5−, Ni ion, 4,4'-bipyridine, and L/D-BCIP (L/D-tert-butoxycarbonyl-2-(imidazole)-1-pyrrolidine). In the structure of Ni-PYI, the 4,4'-bipyridine molecules are connected by Ni ions into 2D layers which are further connected into a 3D rigid framework by L/D-PYI (L/D-pyrrolidin-2-ylimidazole) molecules derived from the decomposition of the L/D-BCIP precursor. The [BW12O40]5− polyoxoanions and the guest molecules are located in the channels of the framework. Two types of Ni-PYI (L or D type) crystals were isolated as enantiomers due to the configuration of chiral PYI molecules in their structures. The compounds ex-Ni-PYI without guest molecules could be easily obtained by soaking the crystals in dichloromethane solution containing diethylamine. The L-type ex-Ni-PYI compound showed high activity for asymmetric dihydroxylation of styrene with excellent enantioselectivity for (R)-phenyl-1,2-ethanediol (ee > 95%) after 60 h reaction at 313 K. The D-type ex-Ni-PYI exhibited similar catalytic activities but gave products with the opposite chirality as shown in Figure 14.
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IBM SPSS Statistics 25 (IBM SPSS Statistics) was used for the statistical analyses. Independent t tests (in the case of unequal variances: Welch’s tests) were conducted to identify any differences between casual users (OPDQ score < 5) and consumers with a SPP-OP use (OPDQ score ≥ 5). These groups were compared regarding Internet usage (h/week), OP usage (h/week) and psychological distress (BSI results). The raw values of the BSI were transformed into standardized T-scores using the available sex-specific norm tables in order to take into account sex-specific variations in reported psychopathological symptoms (Franke, 2000). This permits comparing the BSI results in the context of a standardized T-distribution, which facilitates the interpretation and comparability of the results with population values. Because the group sizes of consumers with a SPP-OP use and casual users differ considerably, we report Hedges g (Sawilowsky, 2009) as measure of effect size. Effects of g = 0.20 are regarded as small, g = 0.50 as medium, and g = 0.80 as large. Because multiple comparisons were conducted, a Bonferroni–Holm correction was applied to control the family-wise error rate (Holm, 1979). To evaluate the risk of common method bias Harman's Single Factor Score was calculated (Harman, 1976; Podsakoff et al., 2003). The test is conducted by loading all relevant variables into one factor in an exploratory factor analysis and then examine the unrotated factor solution. The basic assumption of this test is that common method variance is present when the single factor explains more than 50% of the variance (Podsakoff et al., 2003).
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Macrophages are important phagocytic and antigen-presenting cells. They fulfil the important roles of engulfing the invading fungi and function as the link between innate and adaptive immunity. In Aspergillus fumigatus infection, macrophages inhibit fungal spore germination by forming dense clusters around the spores . Macrophage phagocytosis of fungus into the phagosome is followed by phagosome–lysosome fusion that triggers the generation and release of antimicrobial agents that destroy the fungus . Other macrophages secrete cytokines that attract other inflammatory cells to the infection site . Macrophages, along with other inflammatory cells, form granulomas, which are typical in some fungal infections including cryptococcosis . Like macrophages, dendritic cells (DCs) are antigen-presenting cells but are less efficient in pathogen killing . DCs digest the antigen and present it to naïve T cells, causing their differentiation into different T-helper subsets, including T-helper 1 (Th1) and T-helper-17 (Th17), both of which play critical roles in immunity against fungal disease . Neutrophils are the most abundant of polymorphonuclear cells and the most important for innate antifungal immunity. This underscores the role of neutropenia in the predisposition to IFD . The antifungal property of neutrophils relates to their ability to produce chemokines that are chemoattractants aiding the chemotaxis of inflammatory cells to the infection site and soluble factors with antimicrobial, proteolytic, and nucleolytic properties that damage pathogenic fungi. The hyphal form of fungi is the tissue-invading phenotype. Fungal hyphae may be too large for phagocytosis. Neutrophils produce neutrophil extracellular traps (NET) for the extracellular trapping and killing of fungal hyphae . Natural killer (NK) cells are large lymphocytes that participate in host innate immunity. NK cells cause cytotoxicity by utilizing perforin and granzyme. NK cells also produce cytokines that regulate the function of other immune cells . The antifungal function of NK cells occurs via the damage of fungal hyphae, as seen in infections due to Candida albicans and Aspergillus fumigatus .
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2Review
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In the austenitic phase the NiTi alloy is characterized by a crystal structure with a body-centered cubic lattice that makes it stable at high temperatures and gives the alloy rigidity (high modulus of elasticity) and superelasticity, with very low plasticity characteristics. On the contrary, through a decrease of the temperature below the critical transformation temperature range (TTR) the NiTi alloy changes its crystallographic phase in the twinned martensitic phase, characterized by a closely packed hexagonal lattice (Figure 3). In this form the NiTi alloy shows stability at lower temperature with greater flexibility, cyclic fatigue resistance, softness and ductility, and exhibits the shape memory effect when heated . Instead, the intermediate R-phase does not have a precise crystalline conformation but is rather a hybrid between the two phases described above. It is characterized by the possibility of its atoms to move between adjacent crystalline planes, continuously oscillating between an austenitic and martensitic organization.
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(A, B) patient data including gender and age taken into the study are presented. (C) STK33 immunoreactivity was analyzed using standardized protocols. All cancer samples exhibited “moderate” to “strong” STK33 staining intensity and therefore grouped accordingly. Representative set of CRC is depicted. Asterisk indicates immune cells with strong STK33 immunoreactivity. (D) percentage of colorectal (CRC) and pancreatic (PDAC) tumor specimens classified according to STK33 staining intensity in “moderate” and “strong” is shown. (E) lysates of various cancer cell lines (pancreatic – BxPC3, Capan1, MIA PaCa2; colon – HCT-116, SW480; breast – MDA-MB-231 and lung – A549) were subjected to western blot analysis. Membranes were incubated with STK33-specific antibody. β-actin was used as quality control.
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Mechanical properties of nanoscale thin-films have drawn significant attention in the fields of science and engineering, not only for intellectual reasons but for practical applications in microelectromechanical systems (MEMS) and semiconductor devices. For example, the state-of-the-art three-dimensional memory devices require micro/nanomechanical stability design and simulation in order to prevent structural failures during the microfabrication process, unlike their predecessors based on planar device technologies . During the design, however, properties of bulk materials cannot be used because they frequently deviate from literature values due to the scale effect and also vary depending on process conditions. Therefore, it is essential to develop an evaluation technique that accounts for such material characteristics.
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We thoroughly examined our raw data. We agree that the Pomc(low)/Otp neurons share the common precursors as Pomc (high) / Prdm12 cluster, as shown by the RNA-velocity trajectory analysis in Figure 6A and Figure 6—figure supplement 1B and by the transcriptomic analysis at individual developmental stages (Figure 4—figure supplement 1). We have now addressed this issue in the revised manuscript (Line 528-530).
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However, in rare cases of suspected CTI-dependent flutter which cannot be terminated by ablation despite the presence of a confirmed BB by UHD mapping, alternative diagnosis of intra-isthmus reentry should be ruled out . Very recently, epicardial to endocardial breakthrough at the CTI level has been found in only 4% of the cases, using either conventional mapping (5 patients) , or a UHD system . Concerning posterolateral MI ablation, physicians may also be cautious about rare exceptions which are the endo-epicardial breakthrough circuits propagating through the Marshall bundle epicardial connection for instance . A recent study showed that residual conduction through CS and Marshall tract connections (which could be seen in 51% of the cases after endocardial ablation at the MI level) may be at the origin of apparent UB observation . Another example of endo-epicardial circuit (septo-pulmonary bundle) across a LA roof line with complete endocardial BB is provided in Figure 2 .
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Some neuropsychological profiles could provide some information about the underlying etiology of the MCI (Petersen et al., 1999) and the risk of progression to dementia (Petersen et al., 2001) or regression to normal cognition (Koepsell and Monsell, 2012). However, the information provided solely by the neuropsychological assessment is not enough to give an adequate risk assessment, and therefore some authors suggest complementing this information with the use of biomarkers (Campbell et al., 2013; Dunne et al., 2021). In this context, peripheral biomarkers could add or provide information on the possible etiology of the MCI, the individual risk of progression and help to decide about potential treatments.
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2Review
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Our simulations indicated that homodimers formed preferentially in antiparallel configuration, whereas L18W-PGLa/MG2a formed mainly parallel heterodimers (Fig. 7), which is in agreement with a previous study (52). Interestingly, heterodimer formation has been reported for significantly different lipid bilayers. Thus, dimerization of PGLa and MG2a does not appear to be highly specific to lipid composition, although we cannot comment on the onset of dimerization in other lipid bilayers from our study. The analysis of enthalpic interactions between different amino acids showed that dimer formation is stabilized by salt bridges between MG2a-Glu19 and Lys12 or Lys15 residues of L18W-PGLa (Fig. S22), in agreement with (52). Moreover, there are significant hydrophobic interactions between the peptides. For example, Ulmschneider et al. reported a stabilization of PGLa homodimers by Gly-Ala interactions at high P:L ratios (56). It appears, however, that the sum of all these interactions leads to a preferential formation of L18W-PGLa/MG2a heterodimers as compared to L18W-PGLa/L18W-PGLa or MG2a/MG2a homodimers, which is consistent with previous observations (6).
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In Table 3 the performance of the constituent models on data sets A and B are shown in terms of the micro-F and macro-F measures. The discriminative SVM-based models clearly outperform the probabilistic model (LLDA). More precisely, the Meta-Labeler outperforms all other models in both data sets, exhibiting a notable difference in both metrics compared to them. The prevalence of this method over the other SVM variants, particularly if we take into account the challenging properties of the BioASQ indexing task, suggests that ranking the scores of the different labels for every instance followed by some thresholding strategy is clearly more successful than the traditional classification of instances for every label (i.e. assigning 0 or 1 to an instance for every label). It should be also noted that the Meta-Labeler does not have any particular tuning to cope with the class imbalance (the base models are Vanilla SVMs), opposite to the Tuned SVMs, but still outperforms them. Table 3Performance of component models for the test sets of data sets A and BMicro-FMacro-FModelABABMeta-Labeler0.585550.498530.548840.43381Vanilla SVM0.556750.412540.478910.35355Tuned SVM0.566530.456310.510220.37922LLDA0.369830.388730.301000.37140
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We also examined the relation between activity level and physiologic UE functions to gain insight into the role of different physiologic functions on task performance. The maximal active joint angle, maximal muscle torque, and maximal sEMG amplitude showed high correlations with the activity level, indicating that both the ability to use the full range of motion as well as sufficient muscle strength and activation are needed for adequate task performance. Close monitoring of both muscle function and active range of motion could, therefore, play an important role in starting interventions that minimize functional UE decline in a timely manner. Also, we found a moderate correlation between the PUL score and hand function (i.e., the Nine-Hole Peg Test and Timed TIHM), indicating that both scales measure different aspects of UE function and therefore should be examined separately in clinical practices. This finding is in line with the findings of de Vries et al.,. who found a moderate correlation between the Nine-Hole Peg Test and the Timed TIHM and stated that the tests measure different aspects. They found that the Timed-TIHM evaluates complex patterns, including hand manipulation skills and that the Nine-Hole Peg Test evaluates simple patterns of fine motor coordination, e.g., picking up, placing, and releasing pegs . The occurrence of contractures, i.e., decreased passive range of motion, was relatively uncommon. Hence, it is likely that contractures are not the limiting factor for task performance, which also reflects the fact that we did not find a relation between passive range of motion and UE activity level. In fact, the lack of contractures and occurrence of hypermobility in people with SMA could even have a negative effect on task performance . Although contractures limit the range of motion, they result from tissue changes due to inactivity and muscle weakness, e.g., the infiltration of fatty and connective tissue in the muscle. The stiffening of the tissue, especially at the end range of motion, may allow for better energy storing capacity and the ability to more functionally position the joints .
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Chicken manure compost was collected from the Jiangsu Tianmiaoli Fertilizer Co., Ltd., Long straw-like switchgrass was air dried before transferred into lab, then washed with water and dried at 60°C in oven, which then crushed using a high-speed pulverizer to 40 mesh and oven-dried for further experiments. The chemicals used in this study were purchased from Kelong Chemical Reagent Co., (China), Aladdin (China), or Sigma (United States). The bacterial genomic DNA extraction and plasmid extraction kits were purchased from OMEGA (United States).
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Parallel Mediation Model with path estimates and 95% Confidence Interval. The following variables were included as covariates: individualism, ethnicity, gender, age, occupational status, primary caregiver status, number of parents alive. *p < 0.05; **p < 0.01.
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HPLC chromatogram of QLXZD. (A) Overlapping chromatograms of six batches of QLXZD. (B) Chromatogram at λmax = 210 nm. The reference standards are shown on top, while QLXZD is shown below. (C) Chromatogram at λmax = 254 nm. The reference standards are shown on top, while QLXZD is shown below.
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Sagittal T2 MRI image through the third ventricle, cerebral aqueduct and fourth ventricle showing typical findings for LIAS. Note the convex shape of the tuber cinereum and lamina terminalis, dilated third ventricle, stenosis of the distal aqueduct and normal sized fourth ventricle
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1Other
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For the sample size (N = 10 pairs), we aimed at using the minimum number of subjects that can allow us to carry out meaningful statistical analyses. Using larger numbers of wild animals is not easy to achieve compared with laboratory-based animals. Ethics guidelines often favour minimum samples. Also, given that our study system is female-biased, removing 10 males must have induced some potential disturbances on the sites of capture, wherein these males have lost their harems possibly to other male competitors or to the next dominant female.
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Neutral lipids were estimated by Nile red fluorescence according to Huang et al. (2019). Pellets were resuspended in dimethyl sulfoxide (DMSO) 25% and stained with 1.5 μL Nile red (Sigma-Aldrich, Saint-Quentin Fallavier, France) solution 1 mg mL−1 in DMSO (100%). The 298 K fluorescence at 580 nm was excited at 530 nm (slits 5 nm) (Perkin Elmer LS-55, Villebon sur Yvette, France). DMSO diluted glyceryl trioleate (Sigma-Aldrich T7140-500MG, purity ≥99%) was used for calibration (Supplementary Figure 2).
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A factor loading can be interpreted as a Pearson correlation coefficient between the original variable and the factor. Factor 1 was strongly correlated with Dengue 2 (0.876) and Dengue 3 (0.871). Based on these variables loading highly with Factor 1, and given that Dengue is transmitted by mosquitoes , we classified Factor 1 as a latent variable representing vector-transmitted disease. Factor loadings for ETEC and V. cholerae were strongly correlated with Factor 2 (ETEC = 0.872, V. cholerae = 0.871). Given that ETEC and V. cholerae are transmitted when food or water are contaminated with feces [45, 46], we classified Factor 2 as a latent variable representing food/water-transmitted enteric disease. Lastly, norovirus Norwalk and St. Cloud strains were strongly correlated and loaded most highly with Factor 3 (Norwalk = 0.7426, St. Cloud = 0.7422). Norovirus infection occurs by ingesting stool or vomit from an infected person. Although foodborne and waterborne transmission is possible, norovirus is considered primarily a person-to-person transmitted disease [47–49]; as such, we classified Factor 3 as a person-to-person transmitted enteric disease latent variable.
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In addition to these research papers, three review articles were included in this Special Issue that all discussed epigenetic mechanisms of memory consolidation. Leonetti and colleagues reviewed the role of methylation of the adenosine base (m6A) of RNA in neuronal development and memory formation, emphasizing the overlap in functionality of m6A in these two processes . Importantly, they discuss how m6A-mediated “translational priming” during consolidation could regulate memory locally at synapses. Creighton and colleagues reviewed the epigenetic factors involved in memory consolidation in young, healthy adults and how these mechanisms are altered with age . They conclude by discussing the potential source for age-related changes in the epigenome and how targeting of this has important implications for the development of therapeutic interventions for age-associated memory loss. Finally, Lopez and colleagues review the role of histone modifying and chromatin remodeling enzymes in memory consolidation, highlighting the emerging evidence implicating ATP-dependent chromatin remodeling activity-dependent plasticity in the brain . Additionally, they discuss how chromatin remodeling complexes may contribute to the development of substance use disorders.
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2Review
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Anxiety-like behavior is commonly assessed using the EPM, OFT or light-dark box test. These tests can visualize alterations in locomotor activity, explorative behavior, and risk-taking behavior. The latter can also be assessed using the novelty suppressed feeding test, in which the latency to eat familiar food in an aversive novel environment is indicative for anxiety (Samuels and Hen, 2011). In addition, consumption of ethanol has anxiolytic properties and can as such be used as a measure for anxiety-related behavior (Spanagel et al., 1995).
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In addition to the TVMEM, we postulate a relative risk model for T1i (time to the event of interest) as5\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$ {h}_i(t)={h}_0(t) exp\left\{{\gamma}_1{A}_{1i}+{\gamma}_2{W}_i+\alpha {m}_i(0)\right\} $$\end{document}hit=h0texpγ1A1i+γ2Wi+αmi0where Wi is a vector of the baseline covariates, which could be different from vector Zi in model (1), and h0(⋅) is the baseline risk function. The underlying longitudinal measurement mi(0) at baseline (i.e., at time point t = 0), as approximated by the TVMEM, and at the first-stage intervention A1i are included as predictors in model (5) because the time point at which an individual responds to the first-stage intervention (i.e., T1i) depends only on the type of first-stage intervention the subject received and the baseline characteristics.
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As a consequence, glycobiologists have proposed different graphical representations, with symbols or chemical structures replacing monosaccharides. The description of carbohydrate structures using standard symbolic nomenclature enables easy understanding and communication within the scientific community. Research groups working on carbohydrates have developed schematic depictions with symbols and expansions with greyscale colouring as the so-called Oxford nomenclature (UOXF) [4–5], and even fully coloured schemes later on. Among these, some of the proposed representation forms have been accepted and implemented by several groups and initiatives, namely the Consortium for Functional Glycomics (CFG) . Whereas the initial versions of such representation were limited to mammalian glycans, an extension of the graphical representation of glycans, called SNFG Symbol Nomenclature for Glycans (SNFG) [7–8] resulted from a joint international agreement. The newly proposed nomenclature covers 67 monosaccharides aptly represented in eleven shapes and ten colours. There is the hope that it will cope better with the rapidly growing information on the structure and functions of glycans and polysaccharides from microbes, plants and algae. The rendering of glycan drawing and symbol representations motivated the development of several computer applications using a standardised notation. The earliest glycan editors allowed manual drawing similar to ChemDraw or used input files with glycan sequence KCF (KEGG Chemical Function) in text format for similarity search against other structures deposited in the databases. Later developments supported the construction and representation of glycan structures in symbolic form by computational tools like GlycanBuilder . Since then, several advancements have been made to allow the user to both draw glycans manually or by importing and exporting the structure files in different text formats .
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The study respects all ethical requirements in its objectives and methodologies. We have strictly complied with widely recognized international codes of practice, such as the Nuremberg code, the Helsinki agreement, the conventions of the Council of Europe on human rights and biomedicine, with particular attention to EU legislation: 2001/83/EC, 86/609/EEC, and FP7 Decision nr 1982/2006EC. Human biological samples were required because we needed to test human cells, which have unique biological characteristics, distinct from those of animals. The overall intention of the project was to reduce the number of animal experiments. Only adult patients who were able to give consent were included. All the patients, which were the subjects of our study, gave their consent with regards to scientific treatment and publication of their clinic situation and images. We obtained written informed consent from all patients. This study was approved by our Internal Ethical Committee (CERM committee – 2019/27/Rig1), without any registration in public registry, because this study is not a clinical trial.
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Well waters contained a higher abundance of Peronosporomycetes (Aphanomyces) than aquifer samples. These fungi are known to live in damp soils and aquatic environments and contribute to significant loss of crops due to root rot particularly on legumes72. The higher relative abundance of this taxon within wells may indicate contact with the terrestrial environment and could potentially be linked with landuse and irrigation. Amphitremida (Stramenopile: Labyrinthulomycetes) were present in both well and aquifer samples. Taxa from this group are ecologically significant as decomposers or parasites73. They are well known from marine environments however poorly studied in freshwater. Fungi from the order Pezizomycotina (including the class Dothimdeomycetes) had higher relative abundance in aquifers than wells. This taxon is known to occur within aquifers74, potentially having a significant role in nutrient cycling and biofilm production75.
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Heat map display of murine ABC transporter normalized expression level in five highly purified immunocyte populations based upon ImmGen Deep RNA-seq data (GEO accession: GSE122597), namely NK Cells, Follicular B, Naive CD4+, αβT, γδT cells, and peritoneal macrophages
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a Schematic illustration of the fabrication of PM/Pt microrobots for cell manipulation, anticancer doxorubicin (DOX) drug loading, and delivery. Adapted from Ref. with permission, copyright Wiley Ltd., 2018. b Enzyme-powered nanobots enhance anticancer drug delivery. Adapted from Ref. with permission, copyright Wiley Ltd., 2017. c An overview on the loading and release of doxorubicin on/from n-rGO/Pt micromachines. (1) Physical adsorption of doxorubicin on the outer layer of micromachine (n-rGO). (2) Bubble-propelled swimming of micromachines loaded with doxorubicin in the presence of cancer cells (yellow). (3) The release of doxorubicin upon applying an electrical potential and delivering the drugs to the cancer cells. Adapted from Ref. with permission, copyright Wiley Ltd., 2019. (Color figure online)
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The participants are union leaders who were directly involved in a process aimed at reorganizing two important local federations in Northern Italy. They all have managerial responsibility for work groups but also a role in the steering group at the regional and national level. They are therefore in a position to exert influence on how the guidelines of the national trade union organization are interpreted locally. In the course of the interviews, we asked them to tell us which paths of development they thought the union should focus upon in order to reinforce its capacity to influence the labor market and enhance its image in the eyes of its members.
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We examined only a single record from Argentina, consisting of an adult skull (MACN 33.23) from Tabacal, Orán (Salta). However, Wetzel and Mondolfi mentioned other records from La Victoria and Misión Tacaaglé in Formosa. They also reported a record from Pampa del Indio, Chaco (MACN 30.18), but we identified that specimen as D. s. hybridus.
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Next we used our estimates of time-varying R𝑒𝑓𝑓 to forecast the short-term clinical burden in Australia. Estimates were input into a mathematical model of disease dynamics that was extended to account for imported cases. A sequential Monte Carlo method was used to infer the model parameters and appropriately capture the uncertainty (Moss et al., 2019a), conditional on each of a number of sampled R𝑒𝑓𝑓 trajectories up to 5 April, from which point they were assumed to be constant. The model was subsequently projected forward from April 14 to April 28, to forecast the number of reported cases, assuming a symptomatic detection probability of 80%.
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In 2012, World Health Organization (WHO) recommended that countries should consider pregnant women as a priority group for vaccination . Several studies have shown that maternal influenza immunization could protect pregnant women from severe complications related to influenza virus infection [7–9], and that infants up to 6 months of age from vaccinated women may also benefit [10–13]. For example, in a pooled analysis of three maternal influenza immunization trials in South Africa, Mali and Nepal, there was 20% reduction in all-cause severe clinical pneumonia in infants under 6 months . In addition, a recent systematic review and meta-analysis reported that maternal influenza vaccination was associated with a 48 and 72% reduced risk of infants having laboratory-confirmed influenza infection and associated hospitalization, respectively .
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Genotypes: 3849+10KbC-T/ F508del (n=5), 3849+10KbC-T4382delA (n=1);2789+5G->A /F508del (n=3), 2789+5G->A /M1V (n=2), 2789+5G->A /W1282X (N=2), 2789+5G->A /1602delCT (n=1)D579G /F508del (n=3)D1152H /F508del (n=2), D1152H /D110H (n=1), D1152H /N1303K (n=1)R352Q /Delexon22-23 (n=1), R352Q /F508del (n=1)3272-26A→G /E585X (n=1); R117C /N1303K (n=1)
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Single whole-cell sequencing of microglia presumably circumvents the problems identified by Thrupp et al., and we compared our modules to two recent single-cell studies to further examine the applicability of our findings to the AD autopsy literature. Olah et al. recently identified 9 distinct microglial clusters from a set of AD and non-AD tissue20, and the two most relevant clusters for our findings are shown in Fig. 7 (see Supplementary Data 7 for analysis with all clusters from this paper)20. Cluster 2 is the cluster most enriched for genes from homeostatic groups, which Olah et al. also defined as a homeostatic group using a Naive Bayes classifier. Interestingly, Olah et al. found that cluster 2 is the cluster most enriched for genes that positively correlate with β-amyloid and tau and is also the cluster with the second strongest enrichment for genes that positively associate with clinical AD20. This is consistent with the finding repeated several times in this manuscript that homeostatic genes positively correlate with AD traits in autopsy datasets. The cluster most enriched for DAM stage 2 genes is cluster 5, which they identified as the cluster with the strongest positive association with genes that correlate with clinical AD and the second strongest positive association with genes that correlate with β-amyloid (second to cluster 2). Note that, although clusters 2 and 5 are the clusters most enriched for homeostatic and DAM stage 2 genes, respectively, both clusters are also enriched for some mouse gene lists from other categories (Fig. 7). In general, the clusters from Olah et al. are enriched for a broad range of microglial gene lists from the mouse literature (Supplementary Data 7). This suggests that multiple clusters of microglia contribute to the homeostatic or DAM signal in this tissue, and may partially explain why these gene lists co-correlate in late-stage neocortical AD autopsy tissue (Fig. 5). Alsema et al. also recently profiled single-cell microglia41 and found no relevant differences in microglial composition or gene expression between AD and controls. While this could be for a variety of reasons (as discussed in their paper), we note here that the microglial clusters identified in their manuscript show less overlap with our NPH modules as well as all six mouse microglial gene lists in comparison to the microglial clusters from Olah et al. (see Supplementary Data 7). Thus, the Alsema et al. analysis is less consistent with both our data and the mouse data in comparison to the Olah et al. analysis.Fig. 7Human single-cell microglial RNA-seq data is enriched for our NPH modules and mouse microglial gene lists.Clusters 2 and 5 are from single-cell sequencing of microglia from autopsy tissue (Olah et al.20) and the homeostatic and DAM cluster defining genes from Hasselmann et al.43 are from a xenograph mouse model (see text for details). In each panel, the top two modules in gray (mediumpurple3 and saddlebrown) are NPH modules from this paper; mouse gene lists from Keren-Shaul et al.37 (red) and Mathys et al.38 (blue) are also shown. Cluster 2 from Olah et al. is the cluster from this paper most enriched for homeostatic genes from the mouse literature, and cluster 5 is most enriched for DAM stage 2 genes. Nevertheless, both clusters are also enriched for genes from opposing gene lists (i.e., cluster 2 is enriched for DAM stage 2 genes and cluster 5 is enriched for homeostatic genes), and both are enriched for mediumpurple3. The homeostatic and DAM xenograph clusters are enriched exclusively for mediumpurple3 and saddlebrown, although even here there is some crossover enrichment with mouse lists (i.e., DAM stage 2 gene enrichment with the homeostatic cluster). See main text for discussion and Supplementary Data 7 for all analyses with these datasets, including the analysis displayed in this figure. Each analysis in this figure uses a two-sided Fisher’s exact test and is separately Bonferroni adjusted—dotted line in all panels is p value = 0.05.
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RAW264.7 cells, splenocytes from MOG-reactive TCR transgenic 2D2 mice or lymphocytes from immunized wild-type C57BL/6 mice were cultured under standard conditions and assayed for proliferation using H3-methyl-thymidine incorporation as previously described (Warnecke et al. 2017).
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0Study
12,895
Strengths of our meta-analysis include the strict inclusion criteria, especially restriction to prospective cohort studies, which greatly reduced the likelihood of recall and selection biases. In addition, as individual studies had limited statistical power, our study expanded the sample size and provided more reliable estimates. Finally, the reliability of the findings in sensitivity analyses and Galbraith plot analysis, as well as the significant dose-response relationship, further strengthened our findings.
4
0biomedical
0Study
354,143
XRD investigations revealed that the prepared samples are mixed oxide systems, with typical reflexes in the X-ray diffraction pattern clearly visible for the main components CuO, ZnO, or NiO and ZrO2 (Figure 1). The most notable diffraction peaks in XRD patterns of the samples occurred around 2θ = 32.5°, 35.5°, 38.6°, 48.8°, 61.6°, and 66.3°. According to literature, these peaks can be provided by the monoclinic structure of CuO (JCPDS card No. 48-1548) .
4
0biomedical
0Study
108,536
To address this question, herein, bilateral measurements of popliteal artery flow‐mediated dilation (FMD) were performed to determine endothelial function before and after a 3‐h lying‐down period during which one leg was bent (i.e., 90‐degree angles at the hip and knee) and the contralateral leg remained straight, serving as internal control. The subject was placed in the lying‐down position with the attempt to exclusively examine the effects of arterial bending, independent of gravitational forces naturally occurring with upright sitting. We hypothesized that endothelial dysfunction would be manifested in the popliteal artery of the bent, but not straight, leg. We reasoned that impaired popliteal artery endothelial function in the bent leg would be secondary to the hemodynamic changes subjected to that leg.
4
0biomedical
0Study
154,140
Micronutrients, along with their role in the maintenance and development of physical barriers, assist in enhancing immune function by being involved in the production of antimicrobial proteins and mediation of inflammatory processes . Vitamin and mineral supplements help in neutralizing the damaging effects of a few oxidative agents that can damage cells . Deficiencies of certain nutrients suppress the immune function and increase the susceptibility to various infections [12–14].
4
0biomedical
0Study
35,120
However, Fig. 4b inset shows the K–L plots for Co3O4/CNTs electrode and the slopes of all K–L plots remain approximately constant over the studied potential range. This result indicates the number of electrons transferred in the ORR remained constant. Based on equations (6) and (7), the average n value in ORR was estimated to be 4, suggesting a four-electron (4e−) pathway for electrocatalytic ORR61–63. The ORR dynamics on the CNTs electrode were also investigated by RDE and the average n value in ORR was estimated to be 3.6 from corresponding K–L plots (Figure S4). The jk obtained from the intercept of the K–L plots for the Co3O4/CNTs (16.5 mA cm−2 at 0.8 V) was 3.1-magnitudes larger than that of bare CNTs (5.2 mA cm−2) catalyst and similar to that of Pt/C (17.3 mA cm−2). The ORR activities on as-synthesized Co3O4/CNTs at various pH values were also investigated (Figure S5a). Although the Co3O4/CNTs @ pH 12 has the highest Co3O4 (Table S2), the relatively low Co3+/Co2+ might lead to a high charge-transfer (Figure S5b). Hence, a relatively better electrocatalytic activity was observed at Co3O4/CNTs @ pH 10 in based on the higher jk among all pH-dependent Co3O4/CNTs (Figure S5a inset).
4
0biomedical
0Study
196,771
Besides changes in species composition and functional pathways, the amounts of bacteria in saliva were also a point of focus worth studying as it could influence the stability of oral microbiome as well. To that end, an in-vitro experiment was designed to measure the amounts of bacteria in saliva samples incubated with the 3 beverages and water with a concentration gradient by tracing the changes of optical density (OD) values using the Biolog technology in 4 designate time points from 5 h to 72 h (Fig. 2F and 2G). The average well color development (AWCD) value of each group was calculated using observed value minus that of Group W at the same time point and with the same dilution ratio before statistical analysis commenced (Fig. 2H). In Group T, no significant proliferation was found among AWCD of the 4 timepoints regardless of beverage concentration (P > 0.05). As for the other two groups with interference (orange juice and yoghurt), bacteria commenced proliferating at the beginning and plateaued after 48 h only if the beverage was added to saliva without dilution (P < 0.05); however, when the dilution ratio became higher than 10, no significant proliferation was observed. Based on previous studies’ findings that the concentration of sucrose or citric acid would be diluted for 1,000–10,000 times after 15 min in saliva (Lagerlöf and Dawes, 1985; Bashir et al., 1995), and in combination with our findings of resistance and functional stability of oral microbiome against beverage interference, it was reasonable that lower concentration of beverages had less impact on oral microbiome, but this state might vary if beverages are taken more frequently. This indicated the significance of keeping lower concentration of beverages on maintaining the stable state of oral microbiome, directing a future research orientation to explore the changes of oral microbiome confronting long-term or persistent interference of ingesta, which might result in substantial influence on the characteristics of oral microbiome.
4
0biomedical
0Study
282,883
(11) Predorsal Bones and Scutes. There are ten or eleven predorsal bones with thin anterior and posterior bony expansions. The anterior bones are broader than the posterior ones, and the first three stretch almost vertically (Figs. 2A–2B, 3A) or somewhat postero-ventrally (Fig. 6), whereas the rest are oriented antero-ventrally.
3
0biomedical
0Study
91,812
The pseudo code of RPSNC is shown in Algorithm 1. Two linear programming computations are adopted by Line 14 and Line 20 before and after the equilibrium method, respectively. The linear programming computation before equilibrium aims to obtain the topology, while the one after equilibrium can help to decide when to stop RPSNC. The two linear programming models are the same, but their input complete graphs are different. The balanced relay nodes that are obtained by equilibrium method of Phase II (Line 18 of Algorithm 1) represent local optimization to some extent. Therefore, a retention mechanism that can speedup the convergence is adopted by Line 13. In addition, the value of MINCOSTII will decrease monotonically, which can also help to decide when RPSNC can stop.
1
2other
1Other
352,813
Nine clinical serum specimens were measured by the SBSM. The measured concentration was compared with the concentration provided by the hospital, which was tested through ELISA, Figure 8a,b. The data point closed to the blue dash line means that two results were approximate. The concentration of CA125 measured by the SBSM was slightly higher than the concentration provided by the hospital. The cause of this result may be that the reference curve of CA125 was not exact. However, the total tendency of the result was correct. On the other hand, the concentration of CA15-3 measured by the SBSM fitted to the concentration provided by the hospital. Based on the result presented in Figure 8a,b, the SBSM could be regarded as reliable equipment to test protein concentration. The performance of the SBSM is enough for the POCT.
4
0biomedical
0Study
124,478
Data analyses were conducted using the per-protocol (PP) study population. The PP study population excluded patients who were withdrawn after randomization. For sample size calculation, it was estimated that at a statistical power of 80% at a 5% significance level, 108 patients were required to detect 25% difference in primary end point, where 45% in PVI and 20% in GPA are predicted to have recurrent AF/AT. We enrolled 116 patients to allow for ∼5% not completing follow-up.
4
0biomedical
1Other
151,328
The study population included PD patients followed at the Parkinson and Movement Disorder Program (PMDP) at the University of Alberta between 1999 and 2018, and deceased before 2018. Inclusion criteria included: Diagnosis of PD using UK brain bank criteria (14); followed for at least 5 years. Exclusion criteria included: <5 UPDRS III scores from different years, and no UPDRS-III score in the 2 years prior to time of death and those only had off UPDRS scores. Since we were interested in the trajectory patterns approaching death, five or more assessments and scores recorded close to death were deemed necessary. A flowchart shows original data to the final patient sample (Figure 1).
4
0biomedical
0Study
349,769
Within flavonoids, antioxidant activities differ in relation to the number and positions of hydroxy groups, especially those located on ring B; a catechol or pyrogallol arrangement of ring B enhances the flavonoid antioxidant activity . Moreover, an enolic 3-hydroxy group located on ring C affects remarkably the observable ROS scavenging effects . Accordingly, the presence of a pyrogallol moiety turns myricetin and tricetin (Figure 1) into renowned ROS scavengers.
4
0biomedical
0Study
49,189
We demonstrate our proposed method on a Korean GWAS dataset, which has genotype information of 440,094 SNPs for 435 unrelated Korean patients. The genotypic data and clinical information of the patient were previously collected with written informed consent and with the approval of the Ethics Review Board of the Ajou University Hospital (AJIRB-GEN-GEN-11-304) in the genome-wide association study of AERD . The SNP data were anonymized and then used for an epistasis analysis for this study. The topological properties of the generated networks are examined for identifying statistically significant edges. For further validation, pathway and gene ontology enrichment tests are performed in the Allergy and Asthma Portal (AAP). AAP is built upon InnateDB that is a previously developed integrated analysis platform for innate immune responses.
4
0biomedical
0Study
253,795
The proximity-dependent biotinylation assay, reported here, has captured the association between nsp1 and proteins involved in mRNA processing and stability. The association of nsp1 with the pre-mRNA processing complex is particularly interesting as any role of nsp1 in transcription regulation and maturation of mRNA has not been observed previously (Fig. 1). MERS-CoV nsp1 is known to modulate mRNAs in the nucleus so that only transcribed mRNAs undergo cleavage and degradation (37). Similarly, the influenza A virus endonuclease PA-X, also a host shutoff protein, interacts with multiple subunits of the CPSF complex (38). While other nsps of SARS-CoV-2 have been identified to modulate pre-mRNA splicing (52), our experiment uncovers an association between nsp1 and multiple members of pre-mRNA processing pathways using proximity labeling. Mainly, nsp1 associates with CPSF1, CPSF2, NUDT21, and XRN2, proteins that engage with the RNA polymerase II to induce the termination of pre-mRNA transcription and facilitate pre-mRNA cleavage (Figs. 2 and 4). It is important to note that the proximity-dependent labeling does not distinguish between a stable interaction and a more dynamic transient association.
4
0biomedical
0Study
133,572
The fatigue properties of laser-additive manufactured Ti6Al4V were investigated by Wycisk et al. . Specimens in stress relieved as well as HIPed conditions were analysed for crack initiation site, mean stress sensitivity and overall fatigue performance. Fatigue tests in tension-compression loading at 59 Hz and 20 kHz in the HCF and VHCF regimes until 109 cycles revealed a clear shift of crack initiation from surface to internal initiation with increasing cycles to failure, regardless of the testing principle. They also stated that no effects of test frequency on life span could be determined.
4
0biomedical
0Study
125,672
Similar to colon cancer, NO has a dual function in the development of pancreatic cancer. NO leads to protective or anti-cancer cells effects exhibiting a profile associated with iNOS expression, which mainly induces a large amount of NO to respond to the stimulations.
2
0biomedical
1Other
81,129
The Health Lens Analysis projects varied in the extent to which they incorporated equity. Similarly, while the new public health legislation itself included equity as a core principle, the focus of its implementation has been, in the case of regional public health planning, more generally on the local social determinants of health rather than equity, and in the case of establishing Public Health Partner Authorities, has been on building collaborative relationships with other agencies. The intent of these relationships is to contribute to key priority areas within the State Public Health Plan. For example, to support implementation of a component of the State Public Health Plan on stronger and healthier communities and neighbourhoods and in line with its own planning priorities, the Planning Department has signed a Public Health Partner Authority agreement with the Health Department with the aim to work with Health and local government to provide quality green public open spaces to support people being active, strengthen their contact with nature, and provide places for them to relax and interact. The population health and equity intentions and outcomes of these collaborative relationships are not yet known.
2
2other
1Other
238,437
The target for the indicator “nonpolio AFP rate in children ≤15 years old” was met at the national level and for all governorates in all years, and even exceeded the WHO-established minimum nonpolio AFP rate. Thus, the sensitivity of AFP surveillance has been good before and after the war. A similar finding was reported from Nigeria, the eastern Mediterranean region, and other WHO regions . The lack of an effect of war on this indicator has been found for other countries in conflict, such as Iraq . Similarly, the indicator “investigation within 48 hours of report of cases” met the target at the national level and in all governorates in all years, and was not affected by war in this study, in line with findings from other countries in conflict, such as Iraq .
3
0biomedical
0Study
83,278
Larval source reduction using reservoir management is also cost-effective and is not based on a large workforce or intensive resources required by other conventional malaria control tools. Keiser et al. reviewed the literature to evaluate the efficacy of larval management (i.e. methods creating temporary unfavorable conditions for mosquito breeding–e.g. water or vegetation management) in reducing malaria morbidity and mortality, and found that the risk of malaria was reduced by 88% in areas that actively applied environmental management tools for malaria control. The World Health Organization has been encouraging countries to use integrated vector control in their rational decision-making process for the optimal use of resources for vector control .
4
0biomedical
2Review
231,476
Before conducting a national survey using a PRP, pilot studies should first be conducted in a subset sample of a specific population with an adequate representation of subgroups [2, 8]. In these pilot studies, several accuracy measures should be used, as well as absolute bias and relative bias. If necessary, a correction factor for the PRP can be calculated to acquire valid estimates of disease prevalence in a specific population [7, 44].
3
0biomedical
0Study
43,875
In short, previous literature and the current study suggest that there is a relationship between dACC and the value of switching to an alternative behavior as opposed to the action actually taken. In the revised manuscript we have tried to express this without using the word “deactivation” which carries entirely inappropriate connotations of an overall decrement in activity during task performance. To take this to the domain of neurotransmitters, an enhanced learning signal in dACC would mean a stronger signal indicating the relative value of information to be learnt for an alternative as opposed to the current behavior. If glutamate increases a signal and GABA decreases it then we should see a larger instance of such a signal as glutamate increases and as GABA decreases. This is exactly what we find.
4
0biomedical
0Study
123,317
Several transcriptional proteins were detected only in either SqCC (32) or ADC (53) tissues. However, most proteins had negligible regulation (0.67–1.50-fold change) in either lung subtype, except for JUNB and ANKLE2 proteins. JUNB is uniquely overexpressed in ADC and it is involved in regulating gene activity following the primary factor response. JUNB can promote cell invasion and angiogenesis in cancer cell carcinoma . ANKLE2 was upregulated only in SqCC (1.5-fold), indicating the unique characteristic of this transcriptional protein in SqCC.
4
0biomedical
0Study
212,097
Expression of 13 S. bicolor GRAS genes under different abiotic stresses (strong UV radiation, flooding, PEG, NaCl, heat and cold treatments) at the seedling stage. Error bars were obtained from three measurements. Lowercase letters above the bars indicate significant differences (α = 0.05, LSD) among the treatments
3
0biomedical
0Study
102,370
Statistical analyses of 16S rRNA gene data were conducted as described by Wu et al. (2012). Alpha diversity indices, including the Chao 1, ACE, Shannon, and Simpson metrics were calculated in addition to Good’s coverage of diversity, and rarefaction analyses which were all assessed on OTU abundances using the Mothur software package3 (Schloss et al., 2009). The weighted UniFrac metric was computed for bacteria and archaea separately. Principal component analysis (PCA) was conducted using the genus-level classifications to evaluate compositional similarity among the microbial communities. A Venn diagram of shared and unique OTUs was generated using Mothur to visualize dissimilarity among microbial communities of the four sediments. Heatmap analysis was also performed to visualize OTU distributions and compare abundances across the four sediment samples. Shared taxa present in all four samples (100% threshold) were defined as the core microbiome.
4
0biomedical
0Study
155,278
This phase of the IS system operational model is presented third in sequence to emphasize that the other 2 phases should be used to the greatest extent possible, for reasons of efficiency, practicality, and timeliness. However, in some situations various forms of IR may be needed as complements during the earlier phases as well. For instance, a PAG workshop to identify bottlenecks in the delivery system may need to be complemented by a focused ethnographic study (46) or other inquiries to ensure that bottlenecks in the user system also are identified. Experience in this initiative reveals the need for a careful and systematic process to identify when and for what purpose IR is needed, as well as the most practical and appropriate methods to be used. This requires a close collaboration and negotiation between implementers and researchers, recognizing that the decision to undertake any form of IR will require ethical approval which could entail significant staff effort and delays, and diminish the time and attention for promoting action based on other steps in the cycle. Knowledge brokers can play an important role in designing and facilitating this decision process to ensure that the IR is relevant, pragmatic, and timely in relation to the needs of implementers, and to facilitate the translation and utilization of findings in collaboration with policy and program actors.
2
2other
1Other
210,919
Figure 4 relates existing lesion‐based biomarkers and brain volume‐based biomarkers to PDDS evaluated over all MS subtypes. The Z score (across the entire cohort) of each biomarker was entered into regression analysis with PDDS as the independent variable. The obtained regression coefficients (β) quantitate magnitude of the imaging biomarker relation to disability and allow comparison between alternative biomarkers. Among the six evaluated imaging biomarkers, lesion volume (Fig. 4A) exhibited the weakest relation to PDDS. BPF (Fig. 4C), followed by GMF (Fig. 4E), thalamic volume (Fig. 4D), and WMF (Fig. 4F), all out‐performed lesion volume.
4
0biomedical
0Study
368,259
A total of 257 genome sequences of IBV strains isolated in China or used as a vaccine were collected from GenBank on 22 January 2018. These sequences were aligned using MAFFT v7.313 , and the consensus nucleotide sequence of spike (S-CON) was constructed by the Jalview program and listed in Supplementary Materials (Figures S1 and S2) . Furthermore, the phylogenetic analysis of S-CON was carried out using MEGA (version 5.2.2).
4
0biomedical
0Study
258,317
We searched Embase via Ovid, MEDLINE via Ovid, and Cochrane Central Register electronic databases from their respective dates of conception through to June 5, 2020, limiting our search to English records and human studies. Our search strategy is comprised of key terms for acne, acne treatments, and microbiome. Our detailed search strategy can be found in Supplemental Tables S1-S3. Cited studies from included studies and relevant review articles were screened for additional studies not included in the original search.
4
0biomedical
0Study
77,207
Eight variables were found to be associated independently with diarrhoeal illness in the final multivariate model. Two were related to the handwashing category (children not washing hands with soap and/or not having a handwashing facility in the home: adjusted odds ratio (aOR) 6); four were related to other exposure within the home (children not exclusively breastfed for their first six months; using a wide-mouthed container to store water; children having eaten food cooked the previous day; and not having a rack for drying dishes in the home: aOR 7–40); and three were related to exposure outside the home (a neighbour having had diarrhoea during the previous two weeks; children sharing a toilet with a diarrhoea patient; and children drinking water from outside the home during the five days before enrolment: aOR 10–40) (Table 4).
4
0biomedical
0Study
381,541
In this work, we use synchrotron X-ray imaging, with high spatial and temporal resolutions, to investigate the pore-scale dynamics during immiscible GI in an oil-wet reservoir rock at subsurface conditions (8 MPa and 60°C). This is the three-phase extension of an analysis of two-phase displacement using the apparatus and experimental methodology applied to a quarry carbonate . First, we characterize the fluid–fluid contact angles and pore occupancy to confirm the hydrophobic nature of the rock surfaces and infer the wettability order of the system. Then, we use fast imaging to examine, in situ, during GI, the evolution of (i) gas connectivity; (ii) direct, double and multiple displacements, events; (iii) water connectivity and trapping; and (iv) spreading layers. Finally, we quantify the change in Minkowski functionals—fluid saturations, interfacial areas and curvatures—with time to provide a complete description of the fluid topology in the pore space, i.e. fluid–fluid connectivity and trapping.
4
0biomedical
0Study
155
To conclude, the results in this study show that the response reduction by the precursor is approximately 20–30%. We found that the reduction is fairly independent of masker type, masker level and probe level. These results support psychoacoustical paradigms that are designed to probe the efferent system as indeed activating that system.
4
0biomedical
0Study
215,363
* Relationship with industry reported by at least 1 person. SCI = statement about conflicts of interest of panel members present; SCIR = statement about conflicts of interest of external peer reviews present; DIR = disclosure of the identities of peer reviews; DSFS = disclosure of the specific sources of funding for all stages of guideline development; DTCO = disclosure the types of COI (financial and nonfinancial) that are relevant to the guidelines; DEMC = disclosure of the evaluation and management of the COI; DADI = disclosure of how to access the declarations of interests; CAD = coronary artery disease; STE-ACS = ST-Elevation Acute Coronary Syndromes; NSTE-ACS = Non-ST-Elevation Acute Coronary Syndromes; SCAD = Stable Coronary Artery Disease; ACS = Acute Coronary Syndromes; AF = Atrial Fibrillation.
2
0biomedical
1Other
88,794
We have downloaded all DENV-2 E gene sequences with known time (year) and geographical locality (country) of isolation from GenBank (http://www.ncbi.nlm.nih.gov/). Strains used in this study are presented in the following format: two letters ISO country code/strain name/year of isolation. In order to minimize the problem of over-representation of some countries, such as the Puerto Rico, 194 sequences were subsampled from the larger set of 946 sequences by location and time to create a more equitable spatiotemporal distribution, excluding sequences with 100% identity and removing the recombinants detected by RDP, Chimaera, BootStan, GENECONV, MaxChi and SiScan methods implemented in RDP4 program45 to avoid inferential biases. The sequences, which covers a temporal range of 71 years (1944–2014) from 30 countries, were aligned by MAFFT46. The final alignments included 1,485 nucleotides, and were edited manually with Bioedit47. The strain names and accession numbers are listed in Table S4. The specific sampling problem is described in detail in the supporting information Text S1.
4
0biomedical
0Study
293,608
The second profile, described as “problems with psychological distress”, comprising 8.6% of the adolescents, was characterized by high proportions of indicators related to psychological distress. In this group, 100.0% of the adolescents experienced loneliness and 64.4% had anxiety. Adolescents over the age of 15 were the most represented (85.6%). This profile is the only one in which the majority of the adolescents had low socioeconomic status.
2
0biomedical
0Study
175,632
The most common drugs employed are diuretics, betahistine (a mild H1 agonist and strong H3 antagonist, that reduces both endolymphatic pressure and vestibular nuclei activity), and antiemetic or vestibular suppressants (benzodiazepines, anticholinergics, antihistamines, phenothiazines, and ondansetron) . Surgery embraces different choices, depending on the gravity of the illness: intratympanic steroid perfusion, ablative techniques, endolymphatic sac decompression, intratympanic gentamycin perfusion, vestibular neurectomy, and labyrinthectomy . Another potential treatment strategy is intravenous glycerol .
3
0biomedical
1Other
305,693
In the 10-participant 7T in vivo dataset, we again saw strong commissural connections through dorsal IC (Figure 10). In this dataset, the brachium of the IC and the lateral lemniscus shared connectivity patterns, with similar streamline proportions reaching central and external IC nuclei.
4
0biomedical
0Study
180,015
The fluoropyrimidine capecitabine (CAP) and the vinca alkaloid vinorelbine (VNR) have been shown to be at least as active as iv. counterparts. The two drugs have been compared in an European Organisation for Research and Treatment of Cancer (EORTC) prospective Phase II trials in patients pretreated with taxanes and anthacyclines showing equi-activity but a different toxicity profile, being hand foot syndrome and diarrhea more frequent with CAP and neutropenia more incident for VNR . Dose-finding studies of oral VNR plus CAP have consistently shown that the combination is feasible and well tolerated. Severe neutropenia was the dose-limiting toxicity and the recommended doses of VNR and CAP were respectively 80 mg/m2 day 1 and 8 and 1000–1200 mg/m2 bid day 1→14 every 3 weeks .
4
0biomedical
0Study
254,525
Project archiving and image export. A, Previously constructed networks (Fig. 5) can be stored in a database within NEST Desktop or exported to a file for later reloading. An example list of saved, loadable projects is shown. B, The network graph and its description can be captured as screenshot. C, Charts visualizing activity data (Fig. 7) allow for export to either a rendered image (.png) or a vector graphic (.svg).
2
0biomedical
1Other
245,449
As HNC near the skull base often consists of complex target volumes surrounded by dose-limiting critical organs, highly modulated proton beams with steep dose gradients are required to create optimal treatment plans. This may potentially lead to high LET and increased biological effect in critical structures compared with what is indicated by the fixed RBE weighted dose distribution.6,11 Moreover, our group recently characterized a cohort of patients with skull base HNC with RAIC events after treatment with proton therapy.12 These lesions were overlapping or located just outside the CTV border, indicating a potential increased biological effectiveness of protons due to elevated LET in the dose fall-off area. Therefore, the purpose of the current study was to explore dose and LET correlations with RAIC in a subgroup of patients treated with IMPT for skull base HNC.
4
0biomedical
0Study
46,681
NS1-BP expression was assessed by two independent pathologists who were blinded to patient data. Upon disagreement, the two pathologists reassessed the results until a consensus was reached. A staining index obtained as the intensity of positive staining (moderate low, 1; moderate high, 2; strong, 3) and the proportion of immune-positive cells of interest (0%–25%, 1; 25%–50%, 2; 50%–75%, 3; 75%–100%, 4) was calculated. The two scores were multiplied to yield the total score (1, 2, 3, 4, 6, 8, 9, and 12). Expression level was classified into either low (score 1–4) or high (score 6–12).
4
0biomedical
0Study
393,411
Across the large population‐based studies, ACE‐I/ARBs use was 51–91%, beta‐blockers 52–90%, MRAs 12–56%, and loop diuretics 53–81%. 12 , 13 , 14 , 15 , 17 , 19 In our study, this was 85%, 83%, 40%, and 83%, respectively. Notably, there is a considerably large variation across studies, which might be related to differences in definition. Use of most of the aforementioned medications was particularly lower in the US registries, most likely due to lower guideline adherence. 17 , 19 Prescription rates in our study were comparable with the contemporary Dutch CHECK‐HF registry, a more clearly defined HF population, although we found lower rates for MRAs (40% vs. 56%) and loop diuretics (71% vs. 81%), which can be attributed to focus on HFrEF patients who are possibly more symptomatic (26%, New York Health Association III). 15
4
0biomedical
0Study
299,094
Where i indicates stage and j indicates subnetwork. The SSscore is the stage-specificity score, BCRscore is the breast cancer-relation score, and BCRNRscore is the breast cancer-related non-coding RNA score. All scores are introduced in the following sections. The subnetwork with maximum value was selected as the stage-specific breast cancer-related subnetwork.
4
0biomedical
0Study
270,606
IHC assays were performed using the following antibodies: CDK1, NUSAP1, CEP55, TOP2A, MELK, PBK, RRM2, and MAD2L1 (Affinity Biosciences, dilution 1:200). Allred scores were used to analyze immunohistochemistry. The positive staining intensity (0: negative; 1: weak; 2: moderate; and 3: strong) was multiplied by the staining area (0: <5%; 1: 5–25%; 2: 26–50%; 3: 51–75%; and 4: >75%). The final score (on a scale of 0 to 12) was converted to a scale from 0 to 3 [a score of 0–1 was considered negative (0); 2–4 was considered weakly positive (1); 5–8 was considered medium (2); and 9–12 was considered highly positive (3)]. After immunohistochemical analysis, the sections were scanned to obtain high-resolution (40X) digital images using a 3DHISTECH scanner (Pannoramic, TaiBei) in the pathology laboratory of West China Hospital.
4
0biomedical
0Study
262,480
Materiality conveys a fundamental re-conceptualisation of material things, objects, artefacts and technologies, which are no longer regarded to merely represent or reflect human behaviours and thoughts or to articulate ecological needs, but instead come into view as constituents, stimuli and catalysts of human life and culture themselves (Henare et al. 2007; Carlile and Langley 2013; Lemonnier 2014). Materiality draws attention to the ‘thingness’ of things, the ‘objecthood’ of objects and the ‘artefactuality’ of artefacts, flagging up the possibility of material factors to make a real difference for people’s lives, behaviours and thoughts (Meskell 2005; Jones and Boivin 2018). Broadly conceived, materiality theory seeks to overcome ‘hylomorphism’ (cf. Johnston 2006)—the ancient Greek dualism between ‘form’ (eidos or morphê) and ‘matter’ (hulê)—and the influential Descartian rendering of things as res extensa. Instead, material things are recognised as actively contributing to the makeup of shared lifeworlds and as vital media of thought and action, as much ‘constituting’ as ‘constituted’ (Tilley 2007). Materiality theory acknowledges the agency of things and larger object assemblages and probes into their power to shape human realities and calibrate long-term trajectories of biocultural evolution (Knappett 2005; Boivin 2008; Soentgen 2014).
4
2other
2Review
50,060
The question of whether the EFRs for the older adults showed evidence of increased “neural noise” was also examined in the frequency domain. A mixed-effects model with condition (quiet, SS noise, and AM noise), group (young, old), frequency (F0, H2, and H3), and number of sweeps as fixed factors and participant as a random factor was conducted on the spectral noise floor (calculated across the EFR in response to the stimulus, not the prestimulus baseline). The analysis revealed no significant main effect of group [F(1, 35) = 2.2, p = 0.1] nor significant interactions with group and condition and/or frequency [all p > 0.4].
4
0biomedical
0Study
18,707
These and similar findings that purportedly support movement averaging can all plausibly be explained in terms of participants selecting a single, optimized plan. Conversely, there are many observations that are consistent with the optimization theory, but are difficult to explain in terms of movement averaging. Although optimal behaviour often resembles an averaged movement, there are instances where the optimal solution is to not average. Moving quickly, as we tested here, is one example. Another example is to simply place a virtual barrier in between potential goals to penalize intermediate movements11. In this case, participants easily stop generating intermediate movements and instead move directly to one target or the other. These results demonstrate that intermediate movements are not obligatory, and are therefore unlikely to arise from a low-level movement-averaging process.
4
0biomedical
0Study
186,945
The follow-up evaluation included functional and radiographic assessment, which was carried out by two orthopedic surgeons who were not involved in the operation. The functional assessment consisted of measurements of range of motion (ROM) in the elbow, Visual Analog Scale score (VAS), Elbow Self-Assessment score (ESAS) , Mayo Elbow Performance score (MEPS) and Disabilities of the Arm, Shoulder, and Hand (DASH)Outcome Measure score . The ESAS mainly evaluated the patient’s satisfaction with the elbow using a scale of 1–6: 1, very good; 2, good; 3, satisfied; 4, sufficient; 5, insufficient; and 6, poor.
4
0biomedical
0Study
319,700
Venn diagram of placental microRNAs identified by microRNA profiling. Venn diagram showing the numbers of detected and differentially expressed microRNAs identified in pooled probes from female placentas of each group (Con, control group; N3, intervention group) by qPCR TaqMan low density human microRNA assay. Each pool (n = 1) representing 3 female placentas from each group was screened for 667 human microRNAs. Venn diagram represents the number of separately detected microRNAs in each group and the number of different microRNAs in both groups together (numbers outside of the circles). Total number of different microRNAs are separated in (1) microRNAs detected in one group but not in the other group (numbers exclusively inside the group-specific circle but outside of the intersection and (2) microRNAs common in both groups (numbers in the circle intersection). Within the group of microRNAs common in both groups, the numbers for microRNAs showing differential expression levels between the groups are depicted with upwards and downwards arrow for upregulation and downregulation of microRNAs of the intervention versus control group, respectively. Percent values for respective numbers of detected microRNAs related to 667 microRNAs (total number of screened microRNAs) for each group are shown in parentheses
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65,309
77% of respondents reported obtaining a good fit (accurate impression, comfort and getting enough pressure for orbital growth), cosmesis (symmetry and position of the iris), understanding parents and the child patients' concerns, setting realistic goals and showing compassion as important factors in fitting artificial eyes in children.
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108,687
Sleep and waking activity was measured as previously described . dyrk1aa KO embryos and control WT embryos were raised in a light- and temperature-controlled incubator. Five-day-old larvae were placed in a 24-well plate in the observation chamber of Danio Vision tracking system for acclimation under controlled lighting conditions (12 h–12 h light-dark cycles). Starting from 5 dpf, locomotion of each larva during day and night phases were tracked and analyzed by EthoVision XT7 software over a course of 2 days. Locomotor activity was analyzed by converting raw data into the velocity (cm/s) of each larva per 30-min time-bins.
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For B.1.617.1, 8 mAbs (58, 88, 170, 278, 281, 316, 384, and 398) showed a more than 5-fold reduction in neutralization titers, with most of these showing almost complete knockout of activity (Figure 2A; Table S1). Neutralization of the B.1.617.2 virus, which shares the L452R RBD mutation with B.1.617.1, was measured using an FRNT and compared with the Victoria viral isolate. Neutralization of B.1.617.2 was reduced more than 5-fold for 6 antibodies, neutralization by NTD mAb 159 was lost completely, and neutralization by mAbs 58, 170, 278, 281, and 384 was reduced in common with neutralization of B.1.617.1, suggesting that these antibodies may share an epitope overlapping the RBD L452R substitution. Interestingly, mAb 253 showed increased neutralization of B.1.617.2.Figure 2Neutralization of B.1.617.1 and B.1.617.2 by mAbs(A) Neutralization of B.1.617.1-B and B.1.617.2 by a panel of 20 potent human mAbs. Neutralization of B.1.617.1-B, as measured by pseudovirus assay, is shown as open triangles, and neutralization of B.1.617 virus, as measured by FRNT, is shown as closed circles; comparison is made with neutralization curves for Victoria, which we have generated previously (Supasa et al., 2021). Neutralization titers are reported in Table S1.(B) Equivalent plots for the Vir, Regeneron, AstraZeneca, Lilly, and Adagio antibodies.
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The questionnaire includes questions about typical clinical symptoms of COVID-19, travel history to Wuhan city and other badly affected areas or countries, contact history with confirmed or suspected COVID-19 patients within the past 14 days, etc. (Fig. 1). c)Remind patients to properly put on the mask
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1Other
179,484
We assembled the long reads with Flye (12), using default parameters, to generate chromosome-scale scaffolds. Then, using Minimap2 (13) and SAMtools (14), we mapped the short reads onto the assembled scaffolds to compensate for erroneous bases within the long reads and to create consensus sequences. After polishing of the assembly with Pilon (15), another round of consensus short-read mapping was performed. Then, we removed duplicated contigs and sorted the remainder according to length using Funannotate (16). Finally, we separated the chimeric sequences and performed scaffolding using RaGOO (17) with the Leishmania major Friedlin strain genome (GCA_000002725.2) (18) as a reference guide, aligning all 36 chromosomes for our assembly with the exception of six small contigs totaling 70,152 bp.
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Cartoon representation of the pathways in HIV budding. Multimerisation of Gag with Gag/Pol and the genomic RNA at the plasma membrane causes the change in membrane curvature. Two late domain motifs (PTAP and YPXL) located within the p6 domain of Gag recruit subunits of ESCRT‐I and/or ALIX to the budding site. ESCRT‐II forms as a part of membrane sensing supercomplex and functions through the ALIX pathway. The two pathways converge at the recruitment of CHMP4B and VPS4 for the final scission. ESCRT‐II (together with other early ESCRT proteins) is also a prerequisite for Gag/Pol recruitment. Once the budding neck is sealed, HIV virus‐mediated protease cleavage occurs transforming immature particles to mature infectious virions
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0biomedical
1Other
323,056
As the international guidelines for test adaptations suggested , an iterative procedure to develop the different versions of the questionnaires across languages was followed. In a previous project [see 36 for more information], two psychologists proficient in English and Spanish and with expertise in test adaptation and addictive behaviors translated the most updated version of the scale, the ABS , into English. Then a bilingual teacher unfamiliar with the inventory did a back translation. The analysis of the back translation indicated that the English version could be considered comparable to the original scale. Some minor changes were made in the Castilian Spanish version of the ABS to ensure that all the items were adequate for the Argentinian participants. The same procedure was followed in the project of which this study forms part to adapt the marijuana outcomes from English to Spanish [see 34 for more details]. To adapt the measure to Dutch, research team members, who are bicultural and proficient in English, Spanish and Dutch, with expertise in test adaptations, translated the original English/Spanish versions of the ABS, the personality questionnaires, the mental health measure and the marijuana outcomes questionnaires into Dutch (see below for a detailed description of the scales). Then the research team members compared the versions and, after a thorough discussion, composed a preliminary version of the instruments. Items from the English, Dutch and Spanish ABS versions are presented in S1 Table.
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Applequist pointed out that the Code provided a simple mechanism to save the current use of Astragalus penduliflorus – conservation with a conserved type. As it stood, if Phaca alpina was in synonymy we could know how that name was supposed to be applied. If preponderant usage could change the meaning of the name regardless of what was cited in the protologue, there would be endless disputes and many names would be thrown into doubt. She was concerned that the Committee for Vascular Plants, on which she served, would be getting hundreds of requests to determine the meaning of these names.
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1Other
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In order to stratify the COVID19-positive cohort, we measured seroconversion with multiplex immunoassays detecting IgGs against four different SARS-CoV-2 peptides: full length nucleocapsid, full length spike protein (spike), as well as smaller peptides encompassing the N-terminus domain (NTD) and the receptor-binding domain (RBD) of the spike protein (see Materials and methods). The COVID19 cohort displayed significantly elevated levels of anti-SARS-CoV-2 IgGs in all four assays, with strong inter-individual variability (Figure 1b). As a control, levels of antibodies against the Flu A Hong Kong H3 virus strain were no higher in COVID19 patients (Figure 1—figure supplement 1a). In COVID19 patients, reactivity against the spike protein correlated positively with reactivity against the other three peptides (Figure 1c). Therefore, we generated a seroconversion index by summing Z-scores for each individual seroconversion assay, which enabled us to assign a quantitative seroconversion value to each patient (Figure 1d). For the purpose of this study, we divided the COVID19 cohort into equally sized groups of low and high seroconversion indices, referred hereto as sero-low and sero-high groups, respectively.
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Recent research on the relationship between sucrose import into the vacuole and the accumulation of monosaccharides in leaves lends some support to the idea that the high glucose concentrations in cinv1 cinv2 leaves may be generated by sucrose hydrolysis in the vacuole. Increasing the transport of sucrose into vacuoles (through heterologous expression of the vacuolar sucrose loader TST2.1 from sugar beet) resulted in increased levels of monosaccharides, due to sucrose hydrolysis by vacuolar invertases (Vu et al., 2020). We also considered whether altered expression of genes encoding proteins involved in movement of sugars across the tonoplast and its hydrolysis within the vacuole (Martinoia, 2018; Vu et al., 2020) might contribute to glucose accumulation in the cinv1 cinv2 mutant. No consistent picture emerged. Transcripts for two genes encoding transporters responsible for sugar entry into the vacuole—Tonoplast Sugar Transporter1 (TST1, syn. TMT1; Wormit et al., 2006) and Vacuolar Glucose Transporter (VGT1; Aluri and Büttner, 2007)—were reduced in the mutant, but this was also true of the transcript for the major tonoplast glucose exporter Early Responsive to Dehydration-Like6 (ERDL6; Poschet et al., 2011) (Supplementary Table S5). The transcript for one of the two vacuolar acid invertases, VI2, was also reduced, but transcript levels of VI1, the other vacuolar invertase (Weiszmann et al., 2018), and two further proteins implicated in sugar export (SWEET16 and SUC4; Endler et al., 2006; Klemens et al., 2013) were unaffected (Supplementary Table S5). Further research is required to establish conclusively the reasons for and location of the high glucose contents of cinv1 cinv2 seedling roots, taking into account that both tonoplast sugar transporters and vacuolar invertase activity are regulated at post-translational as well as transcriptional levels (e.g. Wingenter et al., 2011; Vu et al., 2020).
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Effect of employed basis on the isotropic part (Aiso) of HFCs (in MHz) for the coordinating N atoms with B3LYP (model 1 in Fig. 3, optimized without substrate and TPSS-D3/def2-SV(P) and MM region relaxed). CGTOs is the number of contracted Gaussian functions in the basis set.
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We next aim to understand the relationship between the chemistry and the HyBOP model performance for any underlying element. We project the complete cluster test dataset in a 2D principal component (PC) space as shown in Fig. 3a. A popular fingerprinting scheme, SOAP or smooth overlap of atomic positions32, that transforms the structural arrangement of a cluster to a unique vector representation was utilized (see Methods). A larger span in the PC space by an element is characteristic of its higher structural diversity; for example, some of the well-known elements with the most diverse chemical bonding, such as B, C, and S, can be seen to have a much larger PC span. We find a strong correlation of this PC area with the element position in the periodic table; moving down the columns of alkali, alkaline-earth, Group IIIA, Group IVA, Group VA, and Group VIA elements, we note that the configurational diversity systematically decreases. Similarly, moving across the 3d, 4d, and 5d transition rows, the spanned PC area first increases and then decreases in accordance with the number of valence d-electrons. A good match between the expected chemical diversity and the spanned PC area of an element is indicative of a well-sampled and comprehensive (training and test) cluster dataset. Another notable aspect from Fig. 3a is that many of the clusters with high prediction errors lie at the boundary of the PC space which are expected to belong to under-represented regions of the potential energy surface. An improvement in the model performance is thus expected if more clusters from such regions are included in the training dataset. When grouped according to the position in the periodic table, the errors in the model energy prediction roughly evolve as per the element chemistry (see Fig. 3b). Moving down the columns of alkali, alkaline-earth, Group IIIA, Group IVA, Group VA, and Group VIA elements the model prediction errors systematically decrease (within certain approximation), while moving across the 3d, 4d, and 5d transition rows, such errors first increase and then decrease. These trends are consistent with the above discussion on configurational diversity, as captured by the spanned PC area. To further quantify this correlation, the area in the 2D PC space was computed using a convex hull construction and can be seen to match well with the errors in model predictions (Fig. 3b)—with the notable exception of Mo, which had a relatively high model error than expected from the respective configurational diversity value. We argue that developing potentials for elements that show large configurational diversity, or large PC area, should be more difficult. This is because finding a unique set of the HyBOP parameters that capture all such high-energy regions of the energy surface is nontrivial.Fig. 3Trends in model prediction errors with structural diversity of the element.a SOAP fingerprint representation of clusters across different elements as projected along with the first two principal components (PCs). The area covered by an element in the PC space is proportional to its structural diversity, and thus, the difficulty to train an accurate potential model. Also, the structural diversity of an element is closely related to its position in the periodic table. Regions of higher errors mostly occur at the boundary of the PC space which are expected to belong to under-represented configurations. b Trend in the prediction errors for different elements when grouped across different columns or rows of the periodic table. A similar trend can be seen for the elemental PC area, suggesting that elemental structural diversity correlates well with the model errors. The dotted lines capture the approximate trend within a group of elements and guide the eye. c Correlation between the prediction error and the inverse of the smoothness of the element energy surface. d Elements with high prediction errors display either high structural diversity or exhibit highly corrugated energy surfaces. Elements with mean absolute error (MAE) >100 meV/atom have been marked in c and d.
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Glycemia, insulinemia and HOMA index were also evaluated before and after DCI treatment. While no significant differences were found between t0 and t1, a trend towards a decrease was observed for all three outcomes (Figs. 6 and 7). DCI treatment restored glycemia to normal values in 3 out of 4 males with abnormal baseline levels (the individual values decreased from 117, 109 and 110 to 101, 101 and 92, respectively). No significant changes were found for LH and FSH serum levels after DCI treatment, compared to the baseline. (Fig. 8). Fig. 6Effect of D-chiro-inositol (DCI) treatment on glycemia levels. Glycemia levels from 10 male volunteers at baseline (t0) and after 30 days of treatment (t1) with 1 g DCI per day are presented as: a median values of glycemia (box plots) (p = 0.1113; Wilcoxon signed rank sum test). Symbol explanation: see caption of Fig. 1a; b individual glycemia levels (before-after treatment). The graph reports the values for each volunteer participating in the studyFig. 7Effect of D-chiro-inositol (DCI) treatment on insulinemia and HOMA index levels. Insulinemia and HOMA index from 10 male volunteers at baseline (t0) and after 30 days of treatment (t1) with 1 g DCI per day are presented as median values of (a) insulinemia and (b) HOMA index (box plots) (insulinemia, p = 0.3750; HOMA index, p = 0.3594; Wilcoxon signed rank sum test). Symbol explanation: see caption of Fig. 1aFig. 8Effect of D-chiro-inositol (DCI) treatment on Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels. LH and FSH levels from 10 male volunteers at baseline (t0) and after 30 days of treatment (t1) with 1 g DCI per day are presented as median values of (a) LH and (b) FSH levels (box plots) (LH, p = 0.3340; FSH, p = 0.5566; Wilcoxon signed rank sum test). Symbol explanation: see caption of Fig. 1a
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All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.
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Table 3.RINVOQ 30 vs 15 mg review issue input/output and process activitiesInput #DisciplineInput descriptionI1Clinical pharmacologyExposure-response analysisI2Clinical, clinical pharmacology, and statisticsIntegrated safety analysesI3Clinical, clinical pharmacology, and statisticsResults from five phase 3 studies Process activity # Process description (inputs involved) Purposive Integrative nature P1Analysis/assessment of safety at both 15 and 30 mg independently (I1, I2, I3)YesCombinatorialP2Analysis/assessment of benefit at both 15 and 30 mg independently (I1, I3)YesCombinatorialP3Comparative analysis of adverse event profile between 15 and 30 mg doses (I1, I2, I3)YesCombinatorial Output # Description O1FDA approval and labeling for the 15 mg dose
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120,475
Labeled peptide samples from secretome experiments were analyzed by LC–MS/MS using a C-18 reversed phase nano-column (75 µm I.D. × 50 cm, 2 µm particle size, nanoEASY 100 C18: Thermo Fisher Scientific) and a continuous acetonitrile gradient of: 0–30% B in 420 min, 30–43% A in 5 min, and 43–90% B in 1 min (where: A = 0.5% formic acid; B = 90% acetonitrile, 0.5% formic acid). A flow rate of 200 nL/min was used to elute the peptides from the reverse phase nano-column to an emitter nanospray needle for real time ionization and peptide fragmentation in a QExactive Hybrid Quadrupole-Orbitrap mass spectrometer (Thermo Fisher Scientific).
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A–B MLK4 knock-down was induced by doxycycline in HCC1806_sh6 and SUM149PT_sh2 cells grown in non-adherent conditions and subsequently cells were treated with doxorubicin at indicated concentrations for 48 h. The activity of caspases 3/7 was measured using bioluminescence assay. Representative pictures of mammospheres upon doxorubicin treatment are shown. Error bars indicate ±SEM from three independent experiments, performed in triplicates (n = 9). Significance was calculated using an unpaired two-tailed t-test, *p < 0.05, **p < 0.01, ****p < 0.0001. C HCC1806_sh6 cells were injected into mammary fat pads of RAG2−/− mice. Doxycycline was administered one day after the injection to induce MLK4 knock-down. Mice were treated with doxorubicin (4 mg/kg, i.p.) or saline at 5-day intervals, starting from day 7 of the experiment. Tumors were measured twice a week. Error bars indicate ±SEM (n = 10 for control group, n = 8 for doxorubicin-treated group, n = 8 for doxycycline-treated group, n = 10 for combination treatment group). Statistical comparison was performed using two-way ANOVA, *p < 0.05, **p < 0.01, ****p < 0.0001. D Weight of tumors resected at the end of the study. Significance was calculated using one-way ANOVA followed by Tukey multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001.
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