4081 Times Increase in Atmospheric Humidity Capture Capacity via Confinement of 2D Nanosheets in 3D Matrix

Atmospheric humidity capture is urgently needed in humidity-related process. Most of the traditional technology suffers the defects of large energy consumption, low capture capacity, etc. Here, an intelligent gel (IG), which was composed of super hygroscopic materials (SHM, for humidity detection, spontaneous capture and in-situ liquefaction), hydrophilic polymer PEG-6000 (polyethylene glycol, for humidity storage), and hydrophobic polymer PVDF (poly(vinylidene-fluoride), for fast water release), is shown. Based on such kind of integration, effective color responsive humidity detection, spontaneous humidity capture and in-situ liquefaction, under ambient conditions, have been achieved. The synergistic effects between two polymers have given birth to a 3D polymer framework that can shrink upon heating and swelling upon solvent immersion, which then act as a versatile matrix that dispersed evenly the 2D SHM into atomically scale. As a consequence, approximate 4081 times increase in working capacity than the individual SHM has been observed. Being an effective way to manipulate atmospheric humidity at the device level, the present work may open new avenue for next-generational water management systems.

Heterocomplexes between the Atypical Chemokine MIF and the CXC-Motif Chemokine CXCL4L1 Regulate Inflammation and Thrombus Formation

To fulfil their orchestrating function in immune cell trafficking in homeostasis and disease, a network of 49 chemokines and 23 receptors capitalizes on features of specificity, redundancy, and functional selectivity such as biased agonism. The discovery of the chemokine interactome, i.e. heteromeric chemokine-chemokine interactions, even across CC- and CXC-class borders, has further expanded the complexity within the network. Moreover, some inflammatory mediators, which are not structurally linked to classical CC-, CXC-, CX3C-, or C-chemokines, can bind to chemokine receptors and behave as atypical chemokines (ACKs). We identified the cytokine macrophage migration inhibitory factor (MIF) as an ACK that binds to the chemokine receptors CXCR2 and CXCR4 to promote atherogenic leukocyte recruitment. Here, we hypothesized that chemokine-chemokine interactions extend to ACKs and that MIF may form heterocomplexes with classical chemokines. We tested this hypothesis, applying an unbiased chemokine protein binding array. The platelet chemokine CXCL4L1, but not its variant CXCL4 or the CXCR2/CXCR4 ligands CXCL8 or CXCL12, was identified as a candidate interactor. MIF/CXCL4L1 complexation was verified by co-immunoprecipitation, surface plasmon-resonance analysis, and microscale thermophoresis, which also established high-affinity binding (KD=100-150 nM). The binding interface was predicted by peptide array-based mapping and molecular docking. We next determined whether heterocomplex formation modulates inflammatory and atherogenic activities of MIF. MIF-elicited T-cell chemotaxis as assessed in a 3D-matrix-based live cell-imaging set-up was abrogated, when cells were co-incubated with MIF and CXCL4L1. Heterocomplexation also blocked MIF-triggered migration of Egfp+ microglia in cortical cultures in situ. Of note, CXCL4L1 blocked the binding of Alexa-MIF to a soluble ectodomain mimic of CXCR4 and co-incubation with CXCL4L1 attenuated MIF-triggered dynamic mass redistribution in HEK293-CXCR4 transfectants, indicating that complex formation interferes with MIF/CXCR4 pathways. As MIF and CXCL4L1 are abundant platelet products, we finally tested their role in platelet activation. Multi-photon microscopy, FLIM-FRET, and proximity ligation assay visualized heterocomplexes in platelet aggregates and clinical human thrombus sections. Moreover, heterocomplex formation inhibited MIF-stimulated thrombus formation under flow and skewed the morphology of adhering platelets from a large to a small lamellipodia phenotype. Together, our study establishes a novel molecular interaction, adding to the complexity of the chemokine interactome and chemokine/receptor network. MIF/CXCL4L1, or more generally, ACK/CXC-motif chemokine heterocomplexes may be promising target structures to modulate inflammation and thrombosis.

3D Cell Culture Systems: Tumor Application, Advantages, and Disadvantages

The traditional two-dimensional (2D) in vitro cell culture system (on a flat support) has long been used in cancer research. However, this system cannot be fully translated into clinical trials to ideally represent physiological conditions. This culture cannot mimic the natural tumor microenvironment due to the lack of cellular communication (cell-cell) and interaction (cell-cell and cell-matrix). To overcome these limitations, three-dimensional (3D) culture systems are increasingly developed in research and have become essential for tumor research, tissue engineering, and basic biology research. 3D culture has received much attention in the field of biomedicine due to its ability to mimic tissue structure and function. The 3D matrix presents a highly dynamic framework where its components are deposited, degraded, or modified to delineate functions and provide a platform where cells attach to perform their specific functions, including adhesion, proliferation, communication, and apoptosis. So far, various types of models belong to this culture: either the culture based on natural or synthetic adherent matrices used to design 3D scaffolds as biomaterials to form a 3D matrix or based on non-adherent and/or matrix-free matrices to form the spheroids. In this review, we first summarize a comparison between 2D and 3D cultures. Then, we focus on the different components of the natural extracellular matrix that can be used as supports in 3D culture. Then we detail different types of natural supports such as matrigel, hydrogels, hard supports, and different synthetic strategies of 3D matrices such as lyophilization, electrospiding, stereolithography, microfluid by citing the advantages and disadvantages of each of them. Finally, we summarize the different methods of generating normal and tumor spheroids, citing their respective advantages and disadvantages in order to obtain an ideal 3D model (matrix) that retains the following characteristics: better biocompatibility, good mechanical properties corresponding to the tumor tissue, degradability, controllable microstructure and chemical components like the tumor tissue, favorable nutrient exchange and easy separation of the cells from the matrix.

A noval lightsheet scattering microscopy for long-term visualization of cell-ECM interaction

Visualization cell interaction with the extracellular matrix (ECM) mesh works plays a central role in understanding cell behavior and the corresponding regulatory mechanisms by the environment in vivo. However, long term visualization of 3D matrix structures remains challenging mainly due to photobleaching or blind spot in the currently available approaches. In this paper, we developed a label-free method based on light-sheet microcopy, termed light-sheet scattering microscopy (LSSM), as a satisfactory solution to solve this problem. LSSM can reliably determine structure of collagen matrices from different origin including bovine, human and rat tail. We verified that the quality and intensity of collagen structure images acquired by LSSM did not decline with time. LSSM offers abundant wavelength choice for matrix structure, maximizing combination possibilities for fluorescence to label the cells. LSSM can be used for visualizing ECM-cell interaction in 3D for long term and characterization of cell-applied forces. Interestingly, we observed ultrathin thread-like structures between cells and matrix using LSSM, which was not to be seen by normal fluorescence microscopy. In summary, LSSM provides a robust approach to investigate the complex interplay between cells and ECM in vitro under in vivo-mimicking conditions.

Functional assessment of the V390F mutation in the CCTδ subunit of chaperonin containing tailless complex polypeptide 1

The chaperonin containing tailless complex polypeptide 1 (CCT) is a multi-subunit molecular chaperone. It is found in the cytoplasm of all eukaryotic cells, where the oligomeric form plays an essential role in the folding of predominantly the cytoskeletal proteins actin and tubulin. Both the CCT oligomer and monomeric subunits also display functions that extend beyond folding, which are often associated with microtubules and actin filaments. Here, we assess the functional significance of the CCTδ V390F mutation, reported in several cancer cell lines. Upon transfection into B16F1 mouse melanoma cells, GFP-CCTδV390F incorporates into the CCT oligomer more readily than GFP-CCTδ. Furthermore, unlike GFP-CCTδ, GFP-CCTδV390F does not interact with the dynactin complex component, p150Glued. As CCTδ has previously been implicated in altered migration in wound healing assays, we assessed the behaviour of GFP-CCTδV390F and other mutants of CCTδ, previously used to assess functional interactions with p150Glued, in chemotaxis assays. We developed the assay system to incorporate a layer of the inert hydrogel GrowDex® to provide a 3D matrix for chemotaxis assessment and found subtle differences in the migration of B16F1 cells, depending on the presence of the hydrogel.

Integrin β1 Promotes Pancreatic Tumor Growth by Upregulating Kindlin-2 and TGF-β Receptor-2

The tumor microenvironment plays a critical role in defining the growth and malignancy of solid tumors. Extracellular matrix (ECM) proteins such as collagen, vitronectin, and fibronectin are major components of the tumor microenvironment. Tumor growth-promoting reciprocal interaction between ECM and cytoplasmic proteins is regulated by the cell surface receptors called integrins. This study investigated the mechanism by which integrin β1 promotes pancreatic tumor growth. In MIA PaCa-2 pancreatic cancer cell line, the loss of integrin β1 protein reduced the ability of cells to proliferate in a 3D matrix and compromised the ability to form a focal adhesion complex. Decreased expression of integrin α5 was observed in KO cells, which resulted in impaired cell spreading and adhesion on vitronectin and fibronectin. Reduced expression of the integrin-associated protein, kindlin-2 was also recorded. The downregulation of kindlin-2 decreased the phosphorylation of Smad2/3 by reducing the expression of TGF-β receptor 2. These results unravel a new mechanism of integrin β1 in tumor growth by modifying the expression of kindlin-2 and TGF-β receptor 2 signaling.

Three Dimensional Measuring Points Locating Algorithm Based Texture-Patched Matrix Completion Algorithm for Indoor 3D REM Design

In this paper, a novel texture-patch transformed (TPT) three dimensional (3D) matrix completion (MC) method has been proposed with the support of novel 3D measuring points (MPs) locating algorithm to generate practical received signal strength (RSS) database assisted indoor 3D radio environment map (REM) of ultra-high frequency (UHF) television (TV)-band. The exploration of TV-band results in TV white and grey space (TV-WS and TV-GS), which are competent resolution to recoup excess data traffic through cognitive radio networks (CRNs) by dynamic spectrum access (DSA) by secondary user (SU). Maximum wireless data traffic generates in indoor and altitude considered exploration of REM achieves high data rate, so selecting interpolation algorithm is important for getting accurate and timely generated REM. Many MC algorithm shows better results than standard interpolation methods. Instead of using layer-by-layer MC algorithm, TPT-MC algorithm could be used through 3D↔2D conversion. Patch size has been considered through symmetric dataset profile. MC criteria based analysis shows TPT-MC algorithm takes lesser no. of MPs than layer-by-layer MC algorithm. Singular value thresholding (SVT) algorithm is used MC algorithm. TPT-SVT shows advantage over layer-by-layer SVT algorithm on RMSE, correlation, best-fit-line and simulation time on same no. of dataset. The result analysis shows that TPT-SVT algorithm is better in RMSE, closest best-fit-line and correlation coefficient than 2D IDW2, 2D K-NN, 2D kriging, TPT-IDW2, TPT-K-NN, TPT-kriging, 3D IDW2 and layer-by-layer SVT algorithm. Computation time of TPT-SVT is better than 3D IDW2 and SVT. TPT-SVT algorithm takes lesser no. of dataset than SVT algorithm for faithful MC.

Optimized Estimation of Leaf Mass per Area with a 3D Matrix of Vegetation Indices

Leaf mass per area (LMA) is a key plant functional trait closely related to leaf biomass. Estimating LMA in fresh leaves remains challenging due to its masked absorption by leaf water in the short-wave infrared region of reflectance. Vegetation indices (VIs) are popular variables used to estimate LMA. However, their physical foundations are not clear and the generalization ability is limited by the training data. In this study, we proposed a hybrid approach by establishing a three-dimensional (3D) VI matrix for LMA estimation. The relationship between LMA and VIs was constructed using PROSPECT-D model simulations. The three-VI space constituting a 3D matrix was divided into cubical cells and LMA values were assigned to each cell. Then, the 3D matrix retrieves LMA through the three VIs calculated from observations. Two 3D matrices with different VIs were established and validated using a second synthetic dataset, and two comprehensive experimental datasets containing more than 1400 samples of 49 plant species. We found that both 3D matrices allowed good assessments of LMA (R2 = 0.76 and 0.78, RMSE = 0.0016 g/cm2 and 0.0017 g/cm2, respectively for the pooled datasets), and their results were superior to the corresponding single Vis, 2D matrices, and two machine learning methods established with the same VI combinations.

Ovarian Cell Encapsulation in an Enzymatically Crosslinked Silk-Based Hydrogel with Tunable Mechanical Properties

An artificial ovary is a promising approach for preserving fertility in prepubertal girls and women who cannot undergo current cryopreservation strategies. However, this approach is in its infancy, due to the possible challenges of creating a suitable 3D matrix for encapsulating ovarian follicles and stromal cells. To maintain the ovarian stromal cell viability and proliferation, as a first step towards developing an artificial ovary, in this study, a double network hydrogel with a high water swelling capacity (swelling index 15–19) was developed, based on phenol conjugated chitosan (Cs-Ph) and silk fibroin (SF) through an enzymatic crosslinking method using horseradish peroxidase. The addition of SF (1%) to Cs (1%) decreased the storage modulus (G’) from 3500 Pa (Cs1) to 1600 Pa (Cs-SF1), and the hydrogels with a rapid gelation kinetic produced a spatially homogeneous distribution of ovarian cells that demonstrated 167% proliferation after 7 days. This new Cs-SF hydrogel benefits from the toughness and flexibility of SF, and phenolic chemistry could provide the potential microstructure for encapsulating human ovarian stromal cells.