Endometrial Epithelial ARID1A Is Required for Uterine Immune Homeostasis during Early Pregnancy

A growing body of work suggests epigenetic dysregulation contributes to endometriosis pathophysiology and female infertility. The chromatin remodeling complex subunit AT-rich interaction domain 1A (ARID1A) must be properly expressed to maintain normal uterine function. Endometrial epithelial ARID1A is indispensable for pregnancy establishment in mice through regulation of endometrial gland function; however, ARID1A expression is decreased in infertile women with endometriosis. We hypothesized that ARID1A performs critical operations in the endometrial epithelium necessary for fertility besides maintaining gland function. To identify alterations in uterine gene expression resulting from loss of epithelial ARID1A, we performed RNA-sequencing analysis on pre-implantation uteri from Ltf^iCre/+Arid1a^f/f and control mice. Differential expression analysis identified 4181 differentially expressed genes enriched for immune-related ingenuity canonical pathways including agranulocyte adhesion and diapedesis and natural killer cell signaling. RT-qPCR confirmed an increase in pro-inflammatory cytokine and macrophage-related gene expression but a decrease in natural killer cell signaling. Immunostaining confirmed a uterus-specific increase in macrophage infiltration. Flow cytometry delineated an increase in inflammatory macrophages and a decrease in uterine dendritic cells in Ltf^iCre/+Arid1a^f/f uteri. These findings demonstrate a role for endometrial epithelial ARID1A in suppressing inflammation and maintaining uterine immune homeostasis, which are required for successful pregnancy and gynecological health.     

Generalized Approach towards Secretion-Based Protein Production via Neutralization of Secretion-Preventing Cationic Substrate Residues

Many heterologous proteins can be secreted by bacterial ATP-binding cassette (ABC) transporters, provided that they are fused with the C-terminal signal sequence, but some proteins are not secretable even though they carry the right signal sequence. The invention of a method to secrete these non-secretable proteins would be valuable both for understanding the secretory physiology of ABC transporters and for industrial applications. Herein, we postulate that cationic “supercharged” regions within the target substrate protein block the secretion by ABC transporters. We also suggest that the secretion of such substrate proteins can be rescued by neutralizing those cationic supercharged regions via structure-preserving point mutageneses. Surface-protruding, non-structural cationic amino acids within the cationic supercharged regions were replaced by anionic or neutral hydrophilic amino acids, reducing the cationic charge density. The examples of rescued secretions we provide include the spike protein of SARS-CoV-2, glutathione-S-transferase, streptavidin, lipase, tyrosinase, cutinase, growth factors, etc. In summary, our study provides a method to predict the secretability and a tool to rescue the secretion by correcting the secretion-blocking regions, making a significant step in understanding the physiological properties of ABC transporter-dependent protein secretion and laying the foundation for the development of a secretion-based protein-producing platform.     

Single-Step Fast Tissue Clearing of Thick Mouse Brain Tissue for Multi-Dimensional High-Resolution Imaging

Recent advances in optical clearing techniques have dramatically improved deep tissue imaging by reducing the obscuring effects of light scattering and absorption. However, these optical clearing methods require specialized equipment or a lengthy undertaking with complex protocols that can lead to sample volume changes and distortion. In addition, the imaging of cleared tissues has limitations, such as fluorescence bleaching, harmful and foul-smelling solutions, and the difficulty of handling samples in high-viscosity refractive index (RI) matching solutions. To address the various limitations of thick tissue imaging, we developed an Aqueous high refractive Index matching and tissue Clearing solution for Imaging (termed AICI) with a one-step tissue clearing protocol that was easily made at a reasonable price in our own laboratory without any equipment. AICI can rapidly clear a 1 mm thick brain slice within 90 min with simultaneous RI matching, low viscosity, and a high refractive index (RI = 1.466), allowing the imaging of the sample without additional processing. We compared AICI with commercially available RI matching solutions, including optical clear agents (OCAs), for tissue clearing. The viscosity of AICI is closer to that of water compared with other RI matching solutions, and there was a less than 2.3% expansion in the tissue linear morphology during 24 h exposure to AICI. Moreover, AICI remained fluid over 30 days of air exposure, and the EGFP fluorescence signal was only reduced to ~65% after 10 days. AICI showed a limited clearing of brain tissue >3 mm thick. However, fine neuronal structures, such as dendritic spines and axonal boutons, could still be imaged in thick brain slices treated with AICI. Therefore, AICI is useful not only for the three-dimensional (3D) high-resolution identification of neuronal structures, but also for the examination of multiple structural imaging by neuronal distribution, projection, and gene expression in deep brain tissue. AICI is applicable beyond the imaging of fluorescent antibodies and dyes, and can clear a variety of tissue types, making it broadly useful to researchers for optical imaging applications.     

Intratracheal Transplantation of Mesenchymal Stem Cells Attenuates Hyperoxia-Induced Microbial Dysbiosis in the Lungs,Brain, and Gut in Newborn Rats

We attempted to determine whether intratracheal (IT) transplantation of mesenchymal stem cells (MSCs) could simultaneously attenuate hyperoxia-induced lung injuries and microbial dysbiosis of the lungs, brain, and gut in newborn rats. Newborn rats were exposed to hyperoxia (90% oxygen) for 14 days. Human umbilical cord blood-derived MSCs (5 × 10⁵) were transplanted via the IT route on postnatal day (P) five. At P14, the lungs were harvested for histological, biochemical, and microbiome analyses. Bacterial 16S ribosomal RNA genes from the lungs, brain, and large intestine were amplified, pyrosequenced, and analyzed. IT transplantation of MSCs simultaneously attenuated hyperoxia-induced lung inflammation and the ensuing injuries, as well as the dysbiosis of the lungs, brain, and gut. In correlation analyses, lung interleukin-6 (IL-6) levels were significantly positively correlated with the abundance of Proteobacteria in the lungs, brain, and gut, and it was significantly inversely correlated with the abundance of Firmicutes in the gut and lungs and that of Bacteroidetes in the lungs. In conclusion, microbial dysbiosis in the lungs, brain, and gut does not cause but is caused by hyperoxic lung inflammation and ensuing injuries, and IT transplantation of MSCs attenuates dysbiosis in the lungs, brain, and gut, primarily by their anti-oxidative and anti-inflammatory effects.     

Improving Lipid Production of Yarrowia lipolytica by the Aldehyde Dehydrogenase-Mediated Furfural Detoxification

Yarrowia lipolytica, the non-conventional yeast capable of high lipogenesis, is a microbial chassis for producing lipid-based biofuels and chemicals from renewable resources such as lignocellulosic biomass. However, the low tolerance of Y. lipolytica against furfural, a major inhibitory furan aldehyde derived from the pretreatment processes of lignocellulosic biomass, has restricted the efficient conversion of lignocellulosic hydrolysates. In this study, the furfural tolerance of Y. lipolytica has been improved by supporting its endogenous detoxification mechanism. Specifically, the endogenous genes encoding the aldehyde dehydrogenase family proteins were overexpressed in Y. lipolytica to support the conversion of furfural to furoic acid. Among them, YALI0E15400p (FALDH2) has shown the highest conversion rate of furfural to furoic acid and resulted in two-fold increased cell growth and lipid production in the presence of 0.4 g/L of furfural. To our knowledge, this is the first report to identify the native furfural detoxification mechanism and increase furfural resistance through rational engineering in Y. lipolytica. Overall, these results will improve the potential of Y. lipolytica to produce lipids and other value-added chemicals from a carbon-neutral feedstock of lignocellulosic biomass.     

Obtaining depth map from segment-based stereo matching using graph cuts

In the paper, the algorithm of segment-based stereo matching using graph cuts is developed for extracting depth information from the stereo image pairs. The first step of the algorithm employs the mean-shift algorithm to segment the reference image, which ensures our method to correctly estimate in large untextured regions and precisely localize depth boundaries, followed by the use of Adaptive Support Weighted Self-Adaptation dissimilarity algorithm (ASW-SelfAd) for the estimation of initial disparity. This is followed by application of Singular Value Decomposition (SVD) in solving the robust disparity plane fitting. In order to ensure reliable pixel sets for the segment, we filter out outliers which contain occlusion region through three main rules, namely; cross-checking, judging reliable area and disparity distance measurement. Lastly, we apply improved clustering algorithm to merge the neighboring segments. The geometrical relationship of adjacent planes such as parallelism and intersection is employed for determination of whether two planes shall be merged. A new energy function is subsequently formulated with the use of graph cuts for the refinement of the disparity map. Finally, the depth information is extracted from the final disparity map. Experimental results on the Middlebury dataset demonstrate that our approach is effective in improving the state of the art.     

Red bean extract reduces inflammation and increases survival in a murine sepsis model

This study used in vitro and in vivo experimental models to investigate the anti-inflammatory potential of the butanol fraction of red bean ethanol extract (BF-RBEE), which contains the biologically active molecule catechin-7-β-d-glucopyranoside. Treatment with BF-RBEE inhibited nitric oxide production in lipopolysaccharide (LPS)-stimulated macrophages through suppression of extracellular signalregulated kinase and IκBα activation. In an in vivo murine sepsis model, oral administration of BF-RBEE improved mouse survival. Specifically, the survival rate of mice injected with LPS was 0 and 40% in ICR and BALB/c mice, respectively, whereas the survival of mice co-treated with BF-RBEE was 100% in both mouse types. This increase in survival with the BF-RBEE administration was correlated with decreased tumor-necrosis factor-α and interferon-γ and increased interleukin-10. Oral administration of BF-RBEE also inhibited total and ovalbumin-specific IgE production in experimental mice. These results suggest the possible usefulness of red beans in the treatment of inflammatory diseases.     

Dieless drawing steel wires using a dielectric heating method and modeling the process dynamics

Modern industries require the production of multi-functional, inorganic, micron-sized metal wires. This study suggests a novel method that could potentially offer a highly efficient dieless drawing technology for manufacturing thin stainless steel fibers. The method is based on a hot-working principle, using microwaves as the heat source and SiC as the susceptor. Experimental trials with a laboratory rig showed that the new system worked effectively for drawing the stainless steel wires and should be able to realize the diameter attenuation with a diameter reduction of up to 21%. The theoretical model describing the deformation behavior of the stainless steel wires in the working zone along with the constitutive equation of Bingham model modified with a power law and Zener–Hollomon parameter turned out to match very good with the actual results of the experiment. The coefficient of variation of the drawn wire diameter increased, as the draw ratio increased, which could be attributed to the occurrence of the narrow necking zone.     

Turbo multiuser detection for differentially modulated CDMA

In this letter, we study differentially modulated, iteratively decoded CDMA. The iterative multiuser receiver proposed consists of an additional soft-input soft-output (SISO) differential decoder, when compared to turbo multiuser detectors for absolutely modulated systems. Algorithms for iterative decoding with and without phase information at the receiver are developed. The resulting turbo receivers with differential modulation outperform coherent receivers with absolute modulation at moderate to high signal to noise ratios due to the interleaver gain associated with recursive inner encoders in serially concatenated encoding structures.