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.     

Topical Capsaicin in Poly(lactic-co-glycolic)acid (PLGA) Nanoparticles Decreases Acute Itch and Heat Pain

Background: Capsaicin, the hot pepper agent, produces burning followed by desensitization. To treat localized itch or pain with minimal burning, low capsaicin concentrations can be repeatedly applied. We hypothesized that alternatively controlled release of capsaicin from poly(lactic-co-glycolic acid) (PLGA) nanoparticles desensitizes superficially terminating nociceptors, reducing burning. Methods: Capsaicin-loaded PLGA nanoparticles were prepared (single-emulsion solvent evaporation) and characterized (size, morphology, capsaicin loading, encapsulation efficiency, in vitro release profile). Capsaicin-PLGA nanoparticles were applied to murine skin and evaluated in healthy human participants (n = 21) for 4 days under blinded conditions, and itch and nociceptive sensations evoked by mechanical, heat stimuli and pruritogens cowhage, β-alanine, BAM8-22 and histamine were evaluated. Results: Nanoparticles (loading: 58 µg capsaicin/mg) released in vitro 23% capsaicin within the first hour and had complete release at 72 h. In mice, 24 h post-application Capsaicin-PLGA nanoparticles penetrated the dermis and led to decreased nociceptive behavioral responses to heat and mechanical stimulation (desensitization). Application in humans produced a weak to moderate burning, dissipating after 3 h. A loss of heat pain up to 2 weeks was observed. After capsaicin nanoparticles, itch and nociceptive sensations were reduced in response to pruritogens cowhage, β-alanine or BAM8-22, but were normal to histamine. Conclusions: Capsaicin nanoparticles could be useful in reducing pain and itch associated with pruritic diseases that are histamine-independent.     

Bias-error reduction in nanometer resolution microscopic motion tracking vision systems

Microscopic Motion Tracking Systems utilize sub-pixel algorithms to achieve close to nanometer precision motion measurement. This paper characterizes the bias-error of template matching image registration techniques which are commonly used to achieve this level of sub-pixel accuracy. An accurate functional expression for such bias error is derived, which shows how the error depends on image content. Furthermore, this expression is directly utilized in a bias reduction scheme that is shown to significantly reduce the bias error. The newly derived expressions and reduction schemes for bias are applied to a Microscopic Vision System that measures micro-object motion with close to nanometer resolution.     

Application of laser-induced breakdown spectroscopy for detecting leakage of boric acid

Recently, many studies have been conducted on developing on-line leak detection techniques in the reactor vessel head of nuclear power plants. One of them, laser-induced breakdown spectroscopy (LIBS), an effective technique of leak detection, which is a kind of atomic emission spectroscopy that uses a highly energetic laser pulse as the excitation source, has been of interest due to the fast and reliable identification of chemical elements for the precipitates formed by the leakage of cooling water containing boric acid. Experimental setup and tests were performed for the boric acid precipitates formed on the steel for various conditions with a Q-switched 532 nm Nd:YAG laser, optical lenses, an Echelle spectrograph and an intensified charge-coupled device detector. The LIBS system could be applied to obtain a spectral line (～249. 2 nm) corresponding to the atomic boron emission line for the precipitates of boric acid formed on low alloy steels, SA508 and SA533, as a substrate. The LIBS technique based on characterizing boric acid deposits might be used for detecting the leakage of cooling water in reactor vessel head of nuclear power plants.     

Mechanical and moisture-dependent swelling properties of compressed Japanese cedar

In this research, five groups of compressed wood specimens were investigated, four made from compressed Japanese cedar (Cryptomeria japonica D. Don) with compression ratios (CR) of 33%, 50%, 67% and 70%, and one without compressing for control purpose. The measurements of moisture-dependent swelling of the specimens were carried out in the R, T and L directions, with typical strain values of 10%, 1.5% and 0.1% respectively corresponding to the CR = 67%. Mechanical property tests of the compressed wood were also undertaken. Material properties obtained cover radial and tangential swelling strain rates, shear moduli in LR, LT and RT planes, Young’s moduli in the L, R, T directions and Poisson ratios in all planes. The Young’s moduli along the longitudinal and radial directions have been increased by over 300% in relation to the CR of 70%.     

Preparation and photoluminescence properties of γ-KCaPO4: Eu phosphors for near UV-based white LEDs

Novel whitish-blue phosphors based on a phosphate host matrix, γ-KCaPO₄: Eu²⁺, were prepared by a conventional solid-state reaction method using slightly phosphorus deficient conditions and their photoluminescence properties were investigated. The concentration quenching process, temperature dependence of the luminescence and decay curve were also investigated. The γ-KCaPO₄: Eu²⁺ phosphor was efficiently excited by UV-Visible light at wavelengths of 200-450 nm and exhibited a bright whitish-blue emission with a maximum peak wavelength of 473 nm. All of these characteristics suggest that the γ-KCaPO₄: Eu²⁺ phosphors combined with red phosphors could be applicable to near UV-based white LEDs, i.e., only two kinds of phosphor powders are needed for the formation of white light.     

Molecular dynamics study on nanotube-resonators with mass migration applicable to both frequency-tuner and data-storage-media

We investigated the cantilevered carbon-nanotube-resonator including electromigratively movable nanoparticle via classical molecular dynamics simulations and continuum model. The change of the effective mass value, which was closely correlated with the position change of the encapsulated nanoparticle, could be regressed by a power function, the resonance frequency of the carbon-nanotube-resonator could be tuned by controlling the nanoparticle’s position, and the possible frequency-shift-ranges then reached 18–85%. The suggested device could be served as a data-storage-media for electromechanical nonvolatile-memory as well as a frequency-tuner.     

Electrospun PEDOT:PSS/PVP nanofibers as the chemiresistor in chemical vapour sensing

Herein, PEDOT:PSS/PVP nanofibers were produced by electrospinning. The presence of PEDOT:PSS in the nanofibers was confirmed by FT-Raman spectroscopy. The applied voltage-dependent diameter of PEDOT:PSS/PVP nanofibers was observed. Also, sensing behaviors of electrospun PEDOT:PSS/PVP nanofibers were explored by measuring its response upon cyclic exposure to organic vapours such as ethanol, methanol, THF, and acetone at room temperature. When PEDOT:PSS/PVP nanofibers were exposed to each solvent, the protic and aprotic solvents resulted in opposite electrical responses. These findings exhibit that electrospun PEDOT:PSS/PVP nanofibers are the promising candidate for the organic vapour sensing material.