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.     

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.     

Spontaneous assimilation of continuous values and temporal information in causal induction.

Existing models of causal induction primarily rely on the contingency between the presence and the absence of a causal candidate and an effect. Yet, classification of observations into these four types of covariation data may not be straightforward because (a) most causal candidates, in real life, are continuous with ambiguous, intermediate values and because (b) effects may unfold after some temporal lag, providing ambiguous contingency information. Although past studies suggested various reasons why ambiguous information may not be used during causal induction, the authors examined whether learners spontaneously use ambiguous information through a process called causal assimilation. In particular, the authors examined whether learners willingly place ambiguous observations into one of the categories relevant to the causal hypothesis, in accordance with their current causal beliefs. In Experiment 1, people's frequency estimates of contingency data reflected that information ambiguous along a continuous quantity dimension was spontaneously categorized and assimilated in a causal induction task. This assimilation process was moderated by the strength of the upheld causal hypothesis (Experiment 2), could alter the overall perception of a causal relationship (Experiment 3), and could occur over temporal sequences (Experiment 4). (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Mobile health monitoring system based on activity recognition using accelerometer

We propose a new method to recognize a user’s activities of daily living with accelerometers and RFID sensor. Two wireless accelerometers are used for classification of five human body states using decision tree, and detection of RFID-tagged objects with hand movements provides additional instrumental activity information. Besides, we apply our activity recognition module to the health monitoring system. We derive linear regressions for each activity by finding the correlations between the attached accelerometers and the expended calories calculated from gas exchange analyzer under different activities. Finally, we can predict the expended calories more efficiently with only accelerometer sensor depend on the recognized activity. We implement our proposed health monitoring module on smart phones for better practical use.     

A study on squeal noise reduction considering the pad shape of the disc brake system for urban railway vehicles

Journal of Mechanical Science and Technology - In this paper, to help reduce the squeal noise produced during the braking of urban railway vehicles, the shape of the disc brake pad was investigated...