A protein-based prognostic model for pancreatic ductal adenocarcinoma: Construction and validation.

BACKGROUND: Probing relevant proteomic biomarkers may facilitate effective pancreatic adenocarcinoma (PDAC) diagnosis, treatment and prevention. Here, we developed a protein-based prognostic model for PDAC by using relevant proteomic biomarkers data from The Cancer Genome Atlas (TCGA). METHODS: We obtained PDAC's proteomic and clinical data from TCGA and used various analytical tools to identify differentially expressed proteins between normal and cancer tissues. We constructed our protein-based prognostic model and confirmed its accuracy using receiver operating characteristic curve and Kaplan-Meier survival analyses. We elucidated clinical factor-signature protein correlations by clinical correlation assessments and protein coexpression networks. We also used immunohistochemistry (protein expression assessment), Gene Set Enrichment Analysis (protein role identification) and CIBERSORT (infiltrating immune cell distribution assessment). RESULTS: CIITA, BRAF_pS445, AR, YTHDF2, IGFBP2 and CDK1_pT14 were identified as PDAC-associated prognostic proteins. All risk scores calculated using our model provided 1-, 3-, 5-year survival probability at 70 % accuracy. The reliability of our model was validated by the GEO as well. In high- and low-risk groups, age, sex, T- and N- stage disparities were significant, and prognostic and coexpressed proteins correlated. PDAC tissues demonstrated significant CDK1_pT14 overexpression but significant BRAF_pS445, YTHDF2, and IGFBP2 underexpression. Downstream proteins of BRAF were validated by IHC. Low-risk tissues demonstrated more naïve B cells, eosinophils, activated NK cells and regulatory T cells, whereas high-risk tissues demonstrated more activated memory T cells, monocytes, neutrophils, dendritic cells and resting NK cells. CONCLUSIONS: Our protein-based prognostic model for PDAC, along with six signature proteins, might aid in predicting PDAC prognosis and therapeutic targets.

Dynamic label-free imaging of lipid droplets and their link to fatty acid and pyruvate oxidation in mouse eggs.

Mammalian eggs generate most of their ATP by mitochondrial oxidation of pyruvate from the surrounding medium or from fatty acids that are stored as triacylglycerols within lipid droplets. The balance between pyruvate and fatty acid oxidation in generating ATP is not established. We have combined coherent anti-Stokes Raman scattering (CARS) imaging with deuterium labelling of oleic acid to monitor turnover of fatty acids within lipid droplets of living mouse eggs. We found that loss of labelled oleic acid is promoted by pyruvate removal but minimised when ß-oxidation is inhibited. Pyruvate removal also causes a significant dispersion of lipid droplets, while inhibition of ß-oxidation causes droplet clustering. Live imaging of luciferase or FAD autofluorescence from mitochondria, suggest that inhibition of ß-oxidation in mouse eggs only leads to a transient decrease in ATP because there is compensatory uptake of pyruvate into mitochondria. Inhibition of pyruvate uptake followed by ß-oxidation caused a similar and successive decline in ATP. Our data suggest that ß-oxidation and pyruvate oxidation contribute almost equally to resting ATP production in resting mouse eggs and that reorganisation of lipid droplets occurs in response to metabolic demand.

The role of ATP in the differential ability of Sr2+ to trigger Ca2+ oscillations in mouse and human eggs.

At fertilization in mice and humans, the activation of the egg is caused by a series of repetitive Ca2+ oscillations which are initiated by phospholipase-C(zeta)ζ that generates inositol-1,4,5-trisphophate (InsP3). Ca2+ oscillations and egg activation can be triggered in mature mouse eggs by incubation in Sr2+ containing medium, but this does not appear to be effective in human eggs. Here, we have investigated the reason for this apparent difference using mouse eggs, and human eggs that failed to fertilize after IVF or ICSI. Mouse eggs incubated in Ca2+-free, Sr2+-containing medium immediately underwent Ca2+ oscillations but human eggs consistently failed to undergo Ca2+ oscillations in the same Sr2+ medium. We tested the InsP3-receptor (IP3R) sensitivity directly by photo-release of caged InsP3 and found that mouse eggs were about 10 times more sensitive to InsP3 than human eggs. There were no major differences in the Ca2+ store content between mouse and human eggs. However, we found that the ATP concentration was consistently higher in mouse compared to human eggs. When ATP levels were lowered in mouse eggs by incubation in pyruvate-free medium, Sr2+ failed to cause Ca2+ oscillations. When pyruvate was added back to these eggs, the ATP levels increased and Ca2+ oscillations were induced. This suggests that ATP modulates the ability of Sr2+ to stimulate IP3R-induced Ca2+ release in eggs. We suggest that human eggs may be unresponsive to Sr2+ medium because they have a lower level of cytosolic ATP.

A primary effect of palmitic acid on mouse oocytes is the disruption of the structure of the endoplasmic reticulum.

Exposure of mouse oocytes to saturated fatty acids (FAs) such as palmitic acid (PA) has been shown to increase lipid content and cause an endoplasmic reticulum (ER) stress response and changes in the mitochondrial redox state. PA can also disrupt Ca2+ stores in other cell types. The links between these intracellular changes, or whether they are prevented by mono-unsaturated FAs such as oleic acid (OA), is unclear. Here, we have investigated the effects of FAs on mouse oocytes, that are maturated in vitro, using coherent anti-Stokes Raman scattering and two-photon fluorescence microscopy. When oocytes were matured in the presence of PA, there were changes in the aggregation pattern and size of lipid droplets that were mitigated by co-incubation in OA. Maturation in PA alone also caused a distinctive disruption of the ER structure. This effect was prevented by incubation of OA with PA. In contrast, maturation of mouse oocytes in medium containing PA was not associated with any significant change in the redox state of mitochondria or the Ca2+ content of intracellular stores. These data suggest that a primary effect of saturated FAs such as PA on oocytes is to disrupt the structure of the ER and this is not due to an effect on the mitochondria or Ca2+ stores.

GAS2-like proteins mediate communication between microtubules and actin through interactions with end-binding proteins.

Crosstalk between the microtubule (MT) and actin cytoskeletons is fundamental to many cellular processes including cell polarisation and cell motility. Previous work has shown that members of the growth-arrest-specific 2 (GAS2) family mediate the crosstalk between filamentous actin (F-actin) and MTs, but the molecular basis of this process remained unclear. By using fluorescence microscopy, we demonstrate that three members of this family, GAS2-like 1, GAS2-like 2 and GAS2-like 3 (G2L1, G2L2 and G2L3, also known as GAS2L1, GAS2L2 and GAS2L3, respectively) are differentially involved in mediating the crosstalk between F-actin and MTs. Although all localise to actin and MTs, only the exogenous expression of G2L1 and G2L2 influenced MT stability, dynamics and guidance along actin stress fibres. Biochemical analysis and live-cell imaging revealed that their functions are largely due to the association of these proteins with MT plus-end-binding proteins that bind to SxIP or SxLP motifs located at G2L C-termini. Our findings lead to a model in which end-binding (EB) proteins play a key role in mediating actin-MT crosstalk.

High-resolution 3D spatial distribution of complex microbial colonies revealed by mass spectrometry imaging.

INTRODUCTION: Bacterial living states and the distribution of microbial colony signaling molecules are widely studied using mass spectrometry imaging (MSI). However, current approaches often treat 3D colonies as flat 2D disks, inadvertently omitting valuable details. The challenge of achieving 3D MSI in biofilms persists due to the unique properties of microbial samples. OBJECTIVES: The study aimed to develop a new biofilm sample preparation method that can realize high-resolution 3D MSI of bacterial colonies to reveal the spatial organization of bacterial colonies. METHODS: This article introduces the moisture-assisted cryo-section (MACS) method, enabling embedding-free sectioning parallel to the growth plane. The MACS method secures intact sections by controlling ambient humidity and slice thickness, preventing molecular delocalization. RESULTS: Combined with matrix-assisted laser desorption ionization mass spectrometry (MALDI)-MSI, the MACS method provides high-resolution insights into endogenic and exogenous molecule distributions in Pseudomonas aeruginosa (P. aeruginosa) biofilms, including isomeric pairs. Moreover, analyzed colonies are revived into 3D models, vividly depicting molecular distribution from inner to outer layers. Additionally, we investigated metabolite spatiotemporal dynamics in multiple colonies, observing changes over time and distinct patterns in single versus merged colonies. These findings shed light on the repel-merge process for multi-colony formation. Furthermore, our study monitored chemical responses inside biofilms after antibiotic treatment, showing increased antibiotic levels in the outer biofilm layer over time while maintaining low levels in the inner region. Moreover, the MACS method demonstrated its universality and applicability to other bacterial strains. CONCLUSION: These results unveil complex cell activities within biofilm colonies, offering insights into microbe communities. The MACS method is universally applicable to loosely packed microorganism colonies, overcoming the limitations of previously reported MSI methods. It has great potential for studying bacterial-infected cancer tissues and artificial organs, making it a valuable tool in microbiological research.

Will R&D make investors more tolerant? Analysis based on the performance forecast of Chinese listed companies.

In the era of innovation dividends, whether investors, as the main participants in the capital market, can tolerate enterprise innovation activities is the key to whether the capital market can help enterprises innovate. This paper takes the listed companies of Shanghai and Shenzhen A-shares in China that disclosed quantitative performance forecasts from 2016 to 2021 as samples, obtains the market reaction of performance forecasts through the event study method and uses them as proxy variables of investors' short-term performance expectations, and uses multiple regression analysis to explore the impact of corporate R&D on investors' short-term performance expectations. The results are as follows: (1) corporate R&D investment significantly reduces investors' short-term performance expectations (that is, investors have a significant tolerance effect on enterprises with higher R&D investment); (2) the increase in the shareholding ratio of institutional investors weakens the tolerance effect; and (3) with the implementation of China's innovation-driven strategy, the tolerance effect of its capital market on enterprise R&D gradually increases, especially for high-tech companies, but has a low tolerance effect on state-owned companies' R&D risk. The results show that investors in China's capital market are not completely rational in their response to corporate R&D, and investors are willing to bear more short-term performance losses for high R&D investment, which is consistent with prospect theory.

3D-Printed Filters for Efficient Heavy Metal Removal from Water Using PLA@CS/HAP Composites.

Chitosan/Hydroxyapatite composites, enriched with relatively active -NH2 and -OH groups, have emerged as promising adsorbents for heavy metal removal. In this study, we harnessed the potential of CS/HAP composites by developing monolithic PLA@CS/HAP filters utilizing 3D printing and freeze-drying techniques. These filters possess both macroscopic and microscopic porous structures, endowing them with exceptional capabilities for removing heavy metals from water. The adsorption properties of CS/HAP composites were explored by varying the dosage, duration, and initial concentrations of copper ions. The maximum adsorption capacity for Cu2+ was determined to be approximately 119+/-1 mg/g at the natural pH and 298 K. Notably, the monolithic PLA@CS/HAP filters demonstrated remarkable efficiency in the removal of copper ions, with 90% of copper ions effectively removed within a mere 2-h period in a cyclic adsorption experiment. Furthermore, the PLA@CS/HAP filters exhibited a robust dynamic Cu2+ removal capacity (80.8% or even better in less than 35 min) in a dynamic adsorption experiment. Importantly, all materials employed in this study were environmentally friendly. In summary, the PLA@CS/HAP filter offers advantages such as ease of preparation, eco-friendliness, versatility, and broad applicability in diverse wastewater treatment scenarios, thereby presenting a significant potential for practical implementation.

All-perfluoropolymer, nonlinear stability-assisted monolithic surface combines topology-specific superwettability with ultradurability.

Developing versatile and robust surfaces that mimic the skins of living beings to regulate air/liquid/solid matter is critical for many bioinspired applications. Despite notable achievements, such as in the case of developing robust superhydrophobic surfaces, it remains elusive to realize simultaneously topology-specific superwettability and multipronged durability owing to their inherent tradeoff and the lack of a scalable fabrication method. Here, we present a largely unexplored strategy of preparing an all-perfluoropolymer (Teflon), nonlinear stability-assisted monolithic surface for efficient regulating matters. The key to achieving topology-specific superwettability and multilevel durability is the geometric-material mechanics design coupling superwettability stability and mechanical strength. The versatility of the surface is evidenced by its manufacturing feasibility, multiple-use modes (coating, membrane, and adhesive tape), long-term air trapping in 9-m-deep water, low-fouling droplet transportation, and self-cleaning of nanodirt. We also demonstrate its multilevel durability, including strong substrate adhesion, mechanical robustness, and chemical stability, all of which are needed for real-world applications.

MDST-DGCN: A Multilevel Dynamic Spatiotemporal Directed Graph Convolutional Network for Pedestrian Trajectory Prediction.

Pedestrian trajectory prediction is an essential but challenging task. Social interactions between pedestrians have an immense impact on trajectories. A better way to model social interactions generally achieves a more accurate trajectory prediction. To comprehensively model the interactions between pedestrians, we propose a multilevel dynamic spatiotemporal digraph convolutional network (MDST-DGCN). It consists of three parts: a motion encoder to capture the pedestrians' specific motion features, a multilevel dynamic spatiotemporal directed graph encoder (MDST-DGEN) to capture the social interaction features of multiple levels and adaptively fuse them, and a motion decoder to produce the future trajectories. Experimental results on public datasets demonstrate that our model achieves state-of-the-art results in both long-term and short-term predictions for both high-density and low-density crowds.

Quantitatively linking morphology and optical response of individual silver nanohedra.

The optical response of metal nanoparticles is governed by plasmonic resonances, which are dictated by the particle morphology. A thorough understanding of the link between morphology and optical response requires quantitatively measuring optical and structural properties of the same particle. Here we present such a study, correlating electron tomography and optical micro-spectroscopy. The optical measurements determine the scattering and absorption cross-section spectra in absolute units, and electron tomography determines the 3D morphology. Numerical simulations of the spectra for the individual particle geometry, and the specific optical set-up used, allow for a quantitative comparison including the cross-section magnitude. Silver nanoparticles produced by photochemically driven colloidal synthesis, including decahedra, tetrahedra and bi-tetrahedra are investigated. A mismatch of measured and simulated spectra is found in some cases when assuming pure silver particles, which is explained by the presence of a few atomic layers of tarnish on the surface, not evident in electron tomography. The presented method tightens the link between particle morphology and optical response, supporting the predictive design of plasmonic nanomaterials.

Microfluidic synthesis as a new route to produce novel functional materials.

By geometrically constraining fluids into the sub-millimeter scale, microfluidics offers a physical environment largely different from the macroscopic world, as a result of the significantly enhanced surface effects. This environment is characterized by laminar flow and inertial particle behavior, short diffusion distance, and largely enhanced heat exchange. The recent two decades have witnessed the rapid advances of microfluidic technologies in various fields such as biotechnology; analytical science; and diagnostics; as well as physical, chemical, and biological research. On the other hand, one additional field is still emerging. With the advances in nanomaterial and soft matter research, there have been some reports of the advantages discovered during attempts to synthesize these materials on microfluidic chips. As the formation of nanomaterials and soft matters is sensitive to the environment where the building blocks are fed, the unique physical environment of microfluidics and the effectiveness in coupling with other force fields open up a lot of possibilities to form new products as compared to conventional bulk synthesis. This Perspective summarizes the recent progress in producing novel functional materials using microfluidics, such as generating particles with narrow and controlled size distribution, structured hybrid materials, and particles with new structures, completing reactions with a quicker rate and new reaction routes and enabling more effective and efficient control on reactions. Finally, the trend of future development in this field is also discussed.

Are textbooks facilitators or barriers for teachers' teaching and instructional change? An investigation of secondary mathematics teachers in Shanghai, China.

In this paper we report on a survey study of teachers' perceptions of how mathematics textbooks, including broadly student books, teacher manuals, and exercise books, facilitate or hinder teachers' teaching in Shanghai educational settings. For the study we established a conceptual framework about teachers' teaching, partly drawing on Shulman's model of pedagogical reasoning and action of teachers. The data were collected from a stratified random sample of 133 mathematics teachers in 13 secondary schools through a questionnaire, and follow-up interviews with 24 of them. The results revealed the following: the textbooks were highly regarded and used by the Shanghai mathematics teachers as facilitators rather than barriers for their teaching and instructional change, and the facilitation was most evident in the process of transformation and comprehension, and least evident in evaluation in teachers' pedagogical reasoning and action; compared with student books and exercise books, teacher manuals played a larger facilitating role in teachers' teaching; and finally, compared with teacher characteristics, school characteristics had a greater influence on the extent to which textbooks played a role of facilitation or hindrance for teachers' instructional practice. Implications and suggestions are given at the end of the paper. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11858-021-01306-6.

"Barcode" cell sensor microfluidic system: Rapid and sample-to-answer antimicrobial susceptibility testing applicable in resource-limited conditions.

Many rapid antimicrobial susceptibility testing (AST) methods have been proposed to contain clinical antimicrobial resistance (AMR) and preserve the effectiveness of remaining antimicrobials. However, far fewer methods have been proposed to test AMR in resource-limited conditions, such as for frequent safety screenings of water/food/public facilities, urgent surveys of massive samples during a pandemic, or AMR tests in low-income countries. Rapid AST methods realized thus far have a variety of drawbacks when used for such surveys, e.g., high cost and the requirement of expensive instruments such as microscopy. A more reasonable strategy would be to screen samples via onsite testing first, and then send any sample suspected to contain AMR bacteria for advanced testing. Accordingly, a cost-efficient AST is demanded, which can rapidly process a large number of samples without using expensive equipment. To this end, current work demonstrates a novel "barcode" cell sensor based on an adaptive linear filter array as a fully automatic and microscope-free method for counting very small volumes of cells (~1.00 × 104 cells without pre-incubation), wherein suspended cells concentrate into microbars with length proportional to the number of cells. We combined this sensor with an on-chip culture approach we had demonstrated for rapid and automated drug exposure and realized a low-cost and resource-independent platform for portable AST, from which results can be obtained simply through a cell phone. This method has a much shorter turnaround time (2-3 h) than that of standard methods (16-24 h). Thanks to its microscopy-free analysis, affordability, portability, high throughput, and user-friendliness, our "barcode" AST system has the potential to fulfill the various demands of AST when advanced facilities are not available, making it a promising new tool in the fight against AMR.

Quantitative optical microspectroscopy, electron microscopy, and modelling of individual silver nanocubes reveal surface compositional changes at the nanoscale.

The optical response of metal nanoparticles is governed by plasmonic resonances, which depend often intricately on the geometry and composition of the particle and its environment. In this work we describe a method and analysis pipeline unravelling these relations at the single nanoparticle level through a quantitative characterization of the optical and structural properties. It is based on correlating electron microscopy with microspectroscopy measurements of the same particle immersed in media of different refractive indices. The optical measurements quantify the magnitude of both the scattering and the absorption cross sections, while the geometry measured in electron microscopy is used for numerical simulations of the cross section spectra accounting for the experimental conditions. We showcase the method on silver nanocubes of nominal 75 nm edge size. The large amount of information afforded by the quantitative cross section spectra and measuring the same particle in two environments, allows us to identify a specific degradation of the cube surface. We find a layer of tarnish, only a few nanometers thick, a fine surface compositional change of the studied system which would be hardly quantifiable otherwise.

Defect-induced activity enhancement of enzyme-encapsulated metal-organic frameworks revealed in microfluidic gradient mixing synthesis.

Mimicking the cellular environment, metal-organic frameworks (MOFs) are promising for encapsulating enzymes for general applications in environments often unfavorable for native enzymes. Markedly different from previous researches based on bulk solution synthesis, here, we report the synthesis of enzyme-embedded MOFs in a microfluidic laminar flow. The continuously changed concentrations of MOF precursors in the gradient mixing on-chip resulted in structural defects in products. This defect-generating phenomenon enables multimodal pore size distribution in MOFs and therefore allows improved access of substrates to encapsulated enzymes while maintaining the protection to the enzymes. Thus, the as-produced enzyme-MOF composites showed much higher (~one order of magnitude) biological activity than those from conventional bulk solution synthesis. This work suggests that while microfluidic flow synthesis is currently underexplored, it is a promising strategy in producing highly active enzyme-MOF composites.

Clinical study of combined mirror and extracorporeal shock wave therapy on upper limb spasticity in poststroke patients.

Mirror therapy is a simple, inexpensive, and patient-oriented method that has been shown to reduce phantom sensations and pain caused by amputation and improve range of motion, speed, and accuracy of arm movement and function. Extracorporeal shock wave therapy (ESWT) is a new, reversible, and noninvasive method for the treatment of spasticity after stroke. To investigate the therapeutic effect of the combination of mirror and extracorporeal shock wave therapy on upper limb spasticity in poststroke patients. We randomly assigned 120 patients into four groups: A, B, C, and D. All groups received conventional rehabilitation training for 30 min per day, five times a week, for 4 weeks. Moreover, participants in groups A, B, and C also added mirror therapy, ESWT, and a combination of mirror and ESWT, respectively, for 20 min per day. Motor recovery and spasticity were measured using Fugl-Meyer assessment and modified Ashworth scale. The differences in the Fugl-Meyer assessment and modified Ashworth scale scores in group C were significantly greater than those of group D at all observed time points after treatment and were significantly greater than those of groups A and B (P<0.05), but no significant differences were observed between groups A and B until 12 months. Upper extremity spasticity was improved by combined mirror and ESWT.

Reliable and reusable whole polypropylene plastic microfluidic devices for a rapid, low-cost antimicrobial susceptibility test.

Using an antimicrobial susceptibility test (AST) as an example, this work demonstrates a practical method to fabricate microfluidic chips entirely from polypropylene (PP) and the benefits for potential commercial use. Primarily caused by the misuse and abuse of antibiotics, antimicrobial resistance (AMR) is a major threat to modern medicine. The AST is a promising technique to help with the optimal use of antibiotics for reducing AMR. However, current phenotypic ASTs suffer from long turnaround time, while genotypic ASTs suffer from low reliability, and both are unaffordable for routine use. New microfluidics based AST methods are rapid but still unreliable as well as costly due to the PDMS chip material. Herein, we demonstrate a convenient method to fabricate whole PP microfluidic chips with high resolution and fidelity. Unlike PDMS chips, the whole PP chips showed better reliability due to their inertness; they are solvent-compatible and can be conveniently reused and recycled, which largely decreases the cost, and are environmentally friendly. We specially designed 3D chambers that allow for quick cell loading without valving/liquid exchange; this new hydrodynamic design satisfies the shear stress requirement for on-chip bacterial culture, which, compared to reported designs for similar purposes, allows for a simpler, more rapid, and high-throughput operation. Our system allows for reliable tracking of individual cells and acquisition of AST results within 1-3 hours, which is among the group of fastest phenotypic methods. The PP chips are more reliable and affordable than PDMS chips, providing a practical solution to improve current culture-based AST and benefiting the fight against AMR through helping doctors prescribe effective, narrow-spectrum antibiotics; they will also be broadly useful for other applications wherein a reliable, solvent-resistant, anti-fouling, and affordable microfluidic chip is needed.