APP substrate ectodomain defines amyloid-ß peptide length by restraining γ-secretase processivity and facilitating product release.

Sequential proteolysis of the amyloid precursor protein (APP) by γ-secretases generates amyloid-ß (Aß) peptides and defines the proportion of short-to-long Aß peptides, which is tightly connected to Alzheimer's disease (AD) pathogenesis. Here, we study the mechanism that controls substrate processing by γ-secretases and Aß peptide length. We found that polar interactions established by the APPC99 ectodomain (ECD), involving but not limited to its juxtamembrane region, restrain both the extent and degree of γ-secretases processive cleavage by destabilizing enzyme-substrate interactions. We show that increasing hydrophobicity, via mutation or ligand binding, at APPC99 -ECD attenuates substrate-driven product release and rescues the effects of Alzheimer's disease-associated pathogenic γ-secretase and APP variants on Aß length. In addition, our study reveals that APPC99 -ECD facilitates the paradoxical production of longer Aßs caused by some γ-secretase inhibitors, which act as high-affinity competitors of the substrate. These findings assign a pivotal role to the substrate ECD in the sequential proteolysis by γ-secretases and suggest it as a sweet spot for the potential design of APP-targeting compounds selectively promoting its processing by these enzymes.

A Chemical Proteomic Strategy Reveals Inhibitors of Lipoate Salvage in Bacteria and Parasites.

The development of novel anti-infectives requires unprecedented strategies targeting pathways which are solely present in pathogens but absent in humans. Following this principle, we developed inhibitors of lipoic acid (LA) salvage, a crucial pathway for the survival of LA auxotrophic bacteria and parasites but non-essential in human cells. An LA-based probe was selectively transferred onto substrate proteins via lipoate protein ligase (LPL) in intact cells, and their binding sites were determined by mass spectrometry. Probe labeling served as a proxy of LPL activity, enabling in situ screenings for cell-permeable LPL inhibitors. Profiling a focused compound library revealed two substrate analogs (LAMe and C3) as inhibitors, which were further validated by binding studies and co-crystallography. Importantly, LAMe exhibited low toxicity in human cells and achieved killing of Plasmodium falciparum in erythrocytes with an EC50 value of 15 µM, making it the most effective LPL inhibitor reported to date.

An internal docking site stabilizes substrate binding to γ-secretase: Analysis by molecular dynamics simulations.

Amyloid precursor protein (APP) is cleaved and processed sequentially by γ-secretase yielding amyloid ß (Aß) peptides of different lengths. Longer Aß peptides are associated with the formation of neurotoxic plaques related to Alzheimer's disease. Based on the APP substrate-bound structure of γ-secretase, we investigated the enzyme-substrate interaction using molecular dynamics simulations and generated model structures that represent the sequentially cleaved intermediates during the processing reaction. The simulations indicated an internal docking site providing strong enzyme-substrate packing interaction. In the enzyme-substrate complex, it is located close to the region where the helical conformation of the substrate is interrupted and continues toward the active site in an extended conformation. The internal docking site consists of two non-polar pockets that are preferentially filled by large hydrophobic or aromatic substrate side chains to stabilize binding. Placement of smaller residues such as glycine can trigger a shift in the cleavage pattern during the simulations or results in destabilization of substrate binding. The reduced packing by smaller residues also influences the hydration of the active site and the formation of a catalytically active state. The simulations on processed substrate intermediates and a substrate G33I mutation offer an explanation of the experimentally observed relative increase of short Aß fragment production for this mutation. In addition, studies on a substrate K28A mutation indicate that the internal docking site opposes the tendency of substrate dissociation due to a hydrophobic mismatch at the membrane boundary caused by K28 during processing and substrate movement toward the enzyme active site. The proposed internal docking site could also be useful for the specific design of new γ-secretase modulators.

Psychometric evaluation of the Informed Consent Process Scale in Chinese.

BACKGROUND: Informed consent is essential for the ethical conduct of clinical research and is a culturally sensitive issue. But, a measurable Chinese version of the scale to evaluate the informed consent process has not yet been explored in the existing literature. RESEARCH OBJECTIVES: This study aimed to develop and psychometrically test the Chinese version of the Informed Consent Process Scale. RESEARCH DESIGN: Back-translation was conducted to develop the Chinese version of the questionnaire. A cross-sectional survey was administered, after which an exploratory factor analysis was conducted. PARTICIPANTS: We recruited a total of 375 participants who had experience in signing an informed consent form within the previous 3 years in Taiwan. ETHICAL CONSIDERATIONS: This study was approved by two Institutional Review Boards and the autonomy of the participants was respected. FINDINGS: The Chinese version of the Informed Consent Process Scale is composed of three factors with 23 items showing evidence of acceptable reliability and validity. Three major factors were extracted and labeled: Factor 1 - 'Understanding of the research', Factor 2 - 'Trust and confidence' and Factor 3 - 'Doubt and uncertainty'. The three factors accounted for is 52.954 of the total variance with Cronbach's α of .917. DISCUSSION AND CONCLUSION: The finding corroborates previous studies showing that participants had too little understanding on the informed consent forms they signed and implied the need to clarify the critical points in clinical research. The psychometric results indicated good internal consistency and validity for this newly constructed instrument, and it was found worthy of conducting further testing and application.

[Stimultaneous determination of multiple bioactive constituents in Panacis Japonici Rhizoma processed by different methods and grey relational analysis].

A method, for determination of saponins, amino acids and nucleosides in Panacis Japonici Rhizoma of ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry (UFLC-QTRAP-MS/MS), was established to investigate the effect of different processing methods on the target components of Panacis Japonici Rhizoma. The chromatographic separation was performed on a XBridgeC18(4.6 mm×100 mm, 3.5 µm) at 30 °C with a gradient elution of 0.1% formic acid solution-0.1% formic acid acetonitrile, and the flow rate was 0.8 mL·min⁻¹, using multiple-reaction monitoring (MRM) mode. The grey relational analysis was adopted for the analysis of different processing samples. The results showed that the thirty-three constituents were in a good linear range and the correlation coefficient was greater than 0.999 0; the precision, repeatability and stability were good; the average recovery rates were between 95.33% and 101.8%, and the relative standard deviations were less than 5%. The result of grey relational analysis showed that the complete rhizomes without peeling, which were adopted for the microwave dried method, had the best quality. The established method was accurate and reliable, which could be used to appraise the quality of Panacis Japonici Rhizoma. Our study may lay the way for the processing method of Panacis Japonici Rhizoma in optimization,normalization and standardization.

[Comparative analysis of multiple index constituents in Ophiopogonis Radix and Liriopes Radix].

An analytical method based on UFLC-QTRAP-MS/MS was established for simultaneous determination of thirty-three components including steroidal saponins, homoisoflavonoids, amino acids and nucleosides in Ophiopogonis Radix. Thirty-three target components of commercial medicinal materials of Maidong were comparative analysis. Synergi™ Hydro-RP 100 column (2.0 mm × 100 mm, 2.5 µm) was used with 0.1% formic acid solution-0.1% formic acid acetonitrile for gradient elution at a flow rate of 0.4 mL·min⁻¹. In addition, multiple reaction monitoring (MRM) mode was employed. The data were comprehensively processed and analyzed with hierarchical clustering analysis(HCA), principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) methods. All components showed good linearity(r>0.999 0) within the tested ranges. The average recoveries were between 96.23%-102.0%, and the relative standard deviation(RSD) were less than 5%. The results showed that there were significant differences in components between Ophiopogonis Radix and Liriopes Radix, with seven components obviously different. This method was useful for providing basis for the comprehensive evaluation and intrinsic quality control of Ophiopogonis Radix and Liriopes Radix , and may provide a new method reference for the identification of Ophiopogonis Radix and Liriopes Radix.

[Simultaneous determination of lignans and organic acids in Schisandrae Chinensis Fructus by UFLC-Q-TRAP-MS/MS].

An analytical method based on UFLC-QTRAP-MS/MS was developed for simultaneous determination of fifteen components including eleven lignans (schizantherin B, schisandrol B, schizandrin C, γ-schisandrin, deoxyschizandrin, schisantherin, schisandrin, schisanhenol, gomisin D, gomisin J, and angeloylgomisin H) and organic acids (S)-malic acid, D(-)-tartaric acid, protocatechuic acid, and quinic acid) in Schisandrae Chinensis Fructus. Samples from different product specifications were evaluated and analyzed. The chromatographic separation was performed on a Synergi™ Hydro-RP 100Å column (2.0 mm×100 mm, 2.5 µm) at 40 °C with a gradient elution by employing 0.1% aqueous formic acid (A)-acetonitrile (B) as the mobile phase, and the flow rate was 0.4 mL·min⁻¹, using an electrospray ionization (ESI) source and multiple reaction monitoring (MRM) mode. Fifteen components were evaluated synthetically by TOPSIS and gray related degree. The results showed that fifteen components had good linearity (r>0.999 90), and the limits of detection were all satisfactory. The average recoveries of standard addition for the compounds were between 95.42 % and 98.86 %, and the relative standard deviations were less than 5%. The greatest difference of ri in grey related degree was 58.1%, whilst the greatest difference of Ci value in TOPSIS method was 94.8%. The results of these two methods showed that the holistic quality of No. 14 sample was the best. The developed method was accurate and reliable, which was suitable for the simultaneous determination of multiple functional substances and able to provide a new basis for the comprehensive assessment and overall control of the quality of Schisandrae Chinensis Fructus.

[Thoughts of omics research on quality formation in Dao-di herbs].

Dao-di herbs have been recognized as "quality models" with a firmly stable status. The formation of Dao-di herbs quality is involved from the genetic inheritance on the molecular level to the metabolic phenotype of final products, and the full material-based biosynthetic pathway remains unknown. In recent years, an increasing variety of omics technologies has provided new methods and ideas for the analysis of complex life systems and are suitable for explanation of quality formation in Dao-di herbs as well. In order to alleviate the scarcity of natural resources and offer scientific guidance of transplanting varieties, achievements of omics in the aspects of Dao-di herbs from genetics to phenotyping, the biosynthetic pathway of secondary metabolites, the interaction with human body and the new methods of quality evaluation have been summarized. It will be a fundamental work for protection and utilization of Chinese medicine resources.

Influence of calcium hydroxide dressing and acid etching on the push-out bond strengths of three luting resins to root canal dentin.

OBJECTIVES: This study aims to investigate the effects of calcium hydroxide (Ca(OH)2) dressing in root canals and the effects of subsequent acid etching on the adhesion of luting resins to root canals. MATERIALS AND METHODS: Root specimens were prepared from extracted human permanent molars. Specimen canals were (1) filled with etch-and-rinse (Nexus® third generation (NX3)) and two self-adhesive (RelyX Unicem, Maxcem Elite) luting resins, respectively; (2) dressed with Ca(OH)2 before Ca(OH)2 removal and luting resin filling; (3) dressed with Ca(OH)2 before Ca(OH)2 removal and post-cementation; or (4) treated as described in item (2) except that the canals were further etched with phosphoric acid before luting resin filling. Push-out bond strengths were measured and analyzed using one-way analysis of variance, and Fisher's multiple comparison tests provided a follow-up comparison among these four canal treatments. Attenuated total reflectance-Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to analyze the specimen surfaces. RESULTS: Ca(OH)2 dressing adversely affected the bond strengths to canal dentin of the three luting resins tested. Acid etching did not increase the bond strengths. Infrared analysis revealed that Ca(OH)2 dressing caused no structural changes on the dentin surface. XPS and SEM analyses revealed Ca(OH)2 remnants as the ultimate chemical cause leading to the decrease in bond strength. CONCLUSIONS: The bond strength of luting resin to dentin was affected by Ca(OH)2 dressing. Acid etching treatment could not increase the bond strength. CLINICAL RELEVANCE: Adhesion of the fiber post to the root canal wall may be compromised after Ca(OH)2 dressing. An effective method for complete removal of Ca(OH)2 dressing or increase of bond strength for luting resin needs to be developed.

Text2Face: Text-based Face Generation with Geometry and Appearance Control.

Recent years have witnessed the emergence of various techniques proposed for text-based human face generation and manipulation. Such methods, targeting bridging the semantic gap between text and visual contents, provide users with a deft hand to turn ideas into visuals via text interface and enable more diversified multimedia applications. However, due to the flexibility of linguistic expressiveness, the mapping from sentences to desired facial images is clearly many-to-many, causing ambiguities during text-to-face generation. To alleviate these ambiguities, we introduce a local-to-global framework with two graph neural networks (one for geometry and the other for appearance) embedded to model the inter-dependency among facial parts. This is based upon our key observation that the geometry and appearance attributes among different facial components are not mutually independent, i.e., the combinations of part-level facial features are not arbitrary and thus do not conform to a uniform distribution. By learning from the dataset distribution and enabling recommendations given partial descriptions of human faces, these networks are highly suitable for our text-to-face task. Our method is capable of generating high-quality attribute-conditioned facial images from text. Extensive experiments have confirmed the superiority and usability of our method over the prior art.

Effect of Fe and Zn co-doping on LiCoPO4 cathode materials for High-Voltage Lithium-Ion batteries.

Lithium cobalt phosphate (LiCoPO4) has great potential to be developed as a cathode material for lithium-ion batteries (LIBs) due to its structural stability and higher voltage platform with a high theoretical energy density. However, the relatively low diffusion of lithium ions still needs to be improved. In this work, Fe and Zn co-doped LiCoPO4: LiCo0.9-xFe0.1ZnxPO4/C is utilized to enhance the battery performance of LiCoPO4. The electrochemical properties of LiCo0.85Fe0.1Zn0.05PO4/C demonstrated an initial capacity of 118 mAh/g, with 93.4 % capacity retention at 1C after 100 cycles, and a good capacity of 87 mAh/g remained under a high current density of 10C. In addition, the diffusion rate of Li ions was investigated, proving the improvement of the materials with doping. The impedance results also showed a smaller resistance of the doped materials. Furthermore, operando X-ray diffraction displayed a good reversibility of the structural transformation, corresponding to cycling stability. This work provided studies of both the electrochemical properties and structural transformation of Fe and Zn co-doped LiCoPO4, which showed that 10 % Fe and 5 % Zn co-doping enhanced the electrochemical performance of LiCoPO4 as a cathode material in LIBs.

Evaluation of a critical appraisal program for clinical nurses: a controlled before-and-after study.

BACKGROUND: Evidence-based practice is important to.clinical health care professionals. Clinical nurses can make informed decisions by applying the best evidence to their care. However, standardized curricula on evidence-based medicine are lacking in nursing education programs. This study evaluated a critical appraisal education program and assessed its value in increasing nurses' knowledge of critical appraisal and confidence in their critical appraisal skills. METHODS: A controlled before-and-after study design was used. The education program integrated lectures,practice, and group discussion. A questionnaire was used to evaluate participants' knowledge and confidence in critical appraisal pre- and postintervention. RESULTS: Participants' knowledge and confidence in critical appraisal improved significantly postintervention(both p < .001 ). CONCLUSION: A 1-day, small group discussion education program can effectively improve nurses' knowledge and confidence in critical appraisal. Educators and administrators may replicate this education program to improve the quality of nursing care

What Makes a Good Protein-Protein Interaction Stabilizer: Analysis and Application of the Dual-Binding Mechanism.

Protein-protein interactions (PPIs) are essential for biological processes including immune reactions and diseases. Inhibition of PPIs by drug-like compounds is a common basis for therapeutic approaches. In many cases the flat interface of PP complexes prevents discovery of specific compound binding to cavities on one partner and PPI inhibition. However, frequently new pockets are formed at the PP interface that allow accommodation of stabilizers which is often as desirable as inhibition but a much less explored alternative strategy. Herein, we employ molecular dynamics simulations and pocket detection to investigate 18 known stabilizers and associated PP complexes. For most cases, we find that a dual-binding mechanism, a similar stabilizer interaction strength with each protein partner, is an important prerequisite for effective stabilization. A few stabilizers follow an allosteric mechanism by stabilizing the protein bound structure and/or increase the PPI indirectly. On 226 protein-protein complexes, we find in >75% of the cases interface cavities suitable for binding of drug-like compounds. We propose a computational compound identification workflow that exploits new PP interface cavities and optimizes the dual-binding mechanism and apply it to 5 PP complexes. Our study demonstrates a great potential for in silico PPI stabilizers discovery with a wide range of therapeutic applications.

DrawingInStyles: Portrait Image Generation and Editing With Spatially Conditioned StyleGAN.

The research topic of sketch-to-portrait generation has witnessed a boost of progress with deep learning techniques. The recently proposed StyleGAN architectures achieve state-of-the-art generation ability but the original StyleGAN is not friendly for sketch-based creation due to its unconditional generation nature. To address this issue, we propose a direct conditioning strategy to better preserve the spatial information under the StyleGAN framework. Specifically, we introduce Spatially Conditioned StyleGAN (SC-StyleGAN for short), which explicitly injects spatial constraints to the original StyleGAN generation process. We explore two input modalities, sketches and semantic maps, which together allow users to express desired generation results more precisely and easily. Based on SC-StyleGAN, we present DrawingInStyles, a novel drawing interface for non-professional users to easily produce high-quality, photo-realistic face images with precise control, either from scratch or editing existing ones. Qualitative and quantitative evaluations show the superior generation ability of our method to existing and alternative solutions. The usability and expressiveness of our system are confirmed by a user study.

How Modulator Binding at the Amyloidß-γ-Secretase Interface Enhances Substrate Binding and Attenuates Membrane Distortion.

Inhibition of γ-secretase, an intramembrane protease, to reduce secretion of Amyloid-ß (Aß) peptides has been considered for treating Alzheimer's disease. However, γ-secretase inhibitors suffer from severe side effects. As an alternative, γ-secretase modulators (GSM) reduce the generation of toxic peptides by enhancing the cleavage processivity without diminishing the enzyme activity. Starting from a known γ-secretase structure without substrate but in complex with an E2012 GSM, we generated a structural model that included a bound Aß43 peptide and studied interactions among enzyme, substrate, GSM, and lipids. Our result suggests that E2012 binding at the enzyme-substrate-membrane interface attenuates the membrane distortion by shielding the substrate-membrane interaction. The model predicts that the E2012 modulation is charge-dependent and explains the preserved hydrogen acceptor and the aromatic ring observed in many imidazole-based GSM. Predicted effects of γ-secretase mutations on E2012 modulation were confirmed experimentally. We anticipate that the study will facilitate the future development of effective GSMs.

Identity-Aware and Shape-Aware Propagation of Face Editing in Videos.

The development of deep generative models has inspired various facial image editing methods, but many of them are difficult to be directly applied to video editing due to various challenges ranging from imposing 3D constraints, preserving identity consistency, ensuring temporal coherence, etc. To address these challenges, we propose a new framework operating on the StyleGAN2 latent space for identity-aware and shape-aware edit propagation on face videos. In order to reduce the difficulties of maintaining the identity, keeping the original 3D motion, and avoiding shape distortions, we disentangle the StyleGAN2 latent vectors of human face video frames to decouple the appearance, shape, expression, and motion from identity. An edit encoding module is used to map a sequence of image frames to continuous latent codes with 3D parametric control and is trained in a self-supervised manner with identity loss and triple shape losses. Our model supports propagation of edits in various forms: I. direct appearance editing on a specific keyframe, II. implicit editing of face shape via a given reference image, and III. existing latent-based semantic edits. Experiments show that our method works well for various forms of videos in the wild and outperforms an animation-based approach and the recent deep generative techniques.

Reference-Based Deep Line Art Video Colorization.

Coloring line art images based on the colors of reference images is a crucial stage in animation production, which is time-consuming and tedious. This paper proposes a deep architecture to automatically color line art videos with the same color style as the given reference images. Our framework consists of a color transform network and a temporal refinement network based on 3U-net. The color transform network takes the target line art images as well as the line art and color images of the reference images as input and generates corresponding target color images. To cope with the large differences between each target line art image and the reference color images, we propose a distance attention layer that utilizes non-local similarity matching to determine the region correspondences between the target image and the reference images and transforms the local color information from the references to the target. To ensure global color style consistency, we further incorporate Adaptive Instance Normalization (AdaIN) with the transformation parameters obtained from a multiple-layer AdaIN that describes the global color style of the references extracted by an embedder network. The temporal refinement network learns spatiotemporal features through 3D convolutions to ensure the temporal color consistency of the results. Our model can achieve even better coloring results by fine-tuning the parameters with only a small number of samples when dealing with an animation of a new style. To evaluate our method, we build a line art coloring dataset. Experiments show that our method achieves the best performance on line art video coloring compared to the current state-of-the-art methods.

LC-NeRF: Local Controllable Face Generation in Neural Radiance Field.

3D face generation has achieved high visual quality and 3D consistency thanks to the development of neural radiance fields (NeRF). However, these methods model the whole face as a neural radiance field, which limits the controllability of the local regions. In other words, previous methods struggle to independently control local regions, such as the mouth, nose, and hair. To improve local controllability in NeRF-based face generation, we propose LC-NeRF, which is composed of a Local Region Generators Module (LRGM) and a Spatial-Aware Fusion Module (SAFM), allowing for geometry and texture control of local facial regions. The LRGM models different facial regions as independent neural radiance fields and the SAFM is responsible for merging multiple independent neural radiance fields into a complete representation. Finally, LC-NeRF enables the modification of the latent code associated with each individual generator, thereby allowing precise control over the corresponding local region. Qualitative and quantitative evaluations show that our method provides better local controllability than state-of-the-art 3D-aware face generation methods. A perception study reveals that our method outperforms existing state-of-the-art methods in terms of image quality, face consistency, and editing effects. Furthermore, our method exhibits favorable performance in downstream tasks, including real image editing and text-driven facial image editing.

Enzyme-substrate hybrid ß-sheet controls geometry and water access to the γ-secretase active site.

γ-Secretase is an aspartyl intramembrane protease that cleaves the amyloid precursor protein (APP) involved in Alzheimer's disease pathology and other transmembrane proteins. Substrate-bound structures reveal a stable hybrid ß-sheet immediately following the substrate scissile bond consisting of ß1 and ß2 from the enzyme and ß3 from the substrate. Molecular dynamics simulations and enhanced sampling simulations demonstrate that the hybrid ß-sheet stability is strongly correlated with the formation of a stable cleavage-compatible active geometry and it also controls water access to the active site. The hybrid ß-sheet is only stable for substrates with 3 or more C-terminal residues beyond the scissile bond. The simulation model allowed us to predict the effect of Pro and Phe mutations that weaken the formation of the hybrid ß-sheet which were confirmed by experimental testing. Our study provides a direct explanation why γ-secretase preferentially cleaves APP in steps of 3 residues and how the hybrid ß-sheet facilitates γ-secretase proteolysis.

Keyframe Control of Music-driven 3D Dance Generation.

For 3D animators, choreography with artificial intelligence has attracted more attention recently. However, most existing deep learning methods mainly rely on music for dance generation and lack sufficient control over generated dance motions. To address this issue, we introduce the idea of keyframe interpolation for music-driven dance generation and present a novel transition generation technique for choreography. Specifically, this technique synthesizes visually diverse and plausible dance motions by using normalizing flows to learn the probability distribution of dance motions conditioned on a piece of music and a sparse set of key poses. Thus, the generated dance motions respect both the input musical beats and the key poses. To achieve a robust transition of varying lengths between the key poses, we introduce a time embedding at each timestep as an additional condition. Extensive experiments show that our model generates more realistic, diverse, and beat-matching dance motions than the compared state-of-the-art methods, both qualitatively and quantitatively. Our experimental results demonstrate the superiority of the keyframe-based control for improving the diversity of the generated dance motions.