Adjusting Heterodimeric Coiled-Coils (K/E Zipper) to Connect Autophagy-Inducing Peptide with Cell-Penetrating Peptide.

A connection of a functional peptide with a cell-penetrating peptide (CPP) used a heterodimeric coiled-coil as a molecular zipper can improve the intracellular delivery and activity of the functional peptide. However, the chain length of the coiled coil required for functioning as the molecular zipper is unknown at present. To solve the problem, we prepared an autophagy-inducing peptide (AIP) that conjugates with the CPP via heterodimeric coiled-coils consisting of 1 to 4 repeating units (K/E zipper; AIP-Kn and En-CPP), and we investigated the optimum length of the K/E zipper for effective intracellular delivery and autophagy induction. Fluorescence spectroscopy showed that K/E zippers with n = 3 and 4 formed a stable 1:1 hybrid (AIP-K3/E3-CPP and AIP-K4/E4-CPP, respectively). Both AIP-K3 and AIP-K4 were successfully delivered into cells by the corresponding hybrid formation with K3-CPP and K4-CPP, respectively. Interestingly, autophagy was also induced by the K/E zippers with n = 3 and 4, more intensively by the former than by the latter. The peptides and K/E zippers used in this study did not show significant cytotoxicity. These results indicate that the effective induction of autophagy occurs via an exquisite balance of the association and dissociation of the K/E zipper in this system.

Stability Study of mRNA-Lipid Nanoparticles Exposed to Various Conditions Based on the Evaluation between Physicochemical Properties and Their Relation with Protein Expression Ability.

Lipid nanoparticles (LNPs) are currently in the spotlight as delivery systems for mRNA therapeutics and have been used in the Pfizer/BioNTech and Moderna COVID-19 vaccines. mRNA-LNP formulations have been indicated to require strict control, including maintenance at fairly low temperatures during their transport and storage. Since it is a new pharmaceutical modality, there is a lack of information on the systematic investigation of how storage and handling conditions affect the physicochemical properties of mRNA-LNPs and their protein expression ability. In this study, using the mRNA-LNPs with standard composition, we evaluated the effects of temperature, cryoprotectants, vibration, light exposure, and syringe aspiration from the vials on the physicochemical properties of nanoparticles in relation to their in vitro/in vivo protein expression ability. Among these factors, storage at -80 °C without a cryoprotectant caused a decrease in protein expression, which may be attributed to particle aggregation. Exposure to vibration and light also caused similar changes under certain conditions. Exposure to these factors can occur during laboratory and hospital handling. It is essential to have sufficient knowledge of the stability of mRNA-LNPs in terms of their physical properties and protein expression ability at an early stage to ensure reproducible research and development and medical care.

Demonstration of a diffraction-based optical diffuser inspired by the Morpho butterfly.

Optical diffusers are widely used in a variety of light sources to create uniform illumination over a wide field of view. Inspired by the diffraction-based light diffusion of the Morpho butterfly, here we demonstrate a novel diffuser which fulfils (i) high transmittance, (ii) wide angular spread, and (iii) low color dispersion. Two-dimensional nanopatterns were designed using optical simulations to enable simple fabrication. By introducing anisotropy into the surface nanopatterns, we achieved control of anisotropic light diffusion, which has been challenging for conventional diffusers. Next, the designed diffuser was implemented over a large area (100 × 100 mm2) via nanoimprint lithography. The obtained diffuser demonstrated a high transmittance of ∼85% and full width at half maximum (FWHM) of >60° with low color dispersion, outperforming conventional diffusers. Since the presented diffuser has the controllable diffusion properties with low light loss, it has many applications including LED lighting, displays, and daylight harvesting systems.

Nonleisure-Time Physical Activity Guidance Following Minor Ischemic Stroke: A Randomized Clinical Trial.

The purpose of this single-masked randomized clinical trial was to examine whether nonleisure-time physical activity guidance (NLTPAG) improves physical activity levels in patients after minor ischemic stroke. Patients who had been hospitalized for minor ischemic stroke in an acute care hospital (National Health Institute Stroke Scale ≤ 5) were randomized to either an NLTPAG group (n = 17) or a leisure-time physical activity guidance group (n = 16). NLTPAG focused on reducing sedentary behavior and increasing the frequency of walking for shopping and household activities to improve physical activity levels in daily life. Physical activity levels significantly improved only in participants in the NLTPAG group (initial assessment: metabolic equivalents of task = 12.6; final assessment: metabolic equivalents of task = 14.8; p = .035, r = .51). These results suggest that NLTPAG may be effective for improving physical activity levels in patients after minor ischemic stroke.

Valgamicin C, a novel cyclic depsipeptide containing the unusual amino acid cleonine, and related valgamicins A, T and V produced by Amycolatopsis sp. ML1-hF4.

In the course of optimizing pargamicin A production in Amycolatopsis sp. ML1-hF4, we discovered novel cyclic depsipeptide compounds in the broth and designated them valgamicins A, C, T and V. The structures of these molecules were determined by spectroscopic studies, advanced Marfey's method and X-ray crystal structural analysis. Valgamicin C contains the extremely rare amino acid cleonine. To our knowledge, this is the first report of a cleonine-containing metabolite from a naturally isolated microorganism without any breeding or mutation treatment. None of the valgamicins showed potent antibacterial activity against either Gram-positive or -negative bacteria. Valgamicins A, C and T exhibited moderate cytotoxicity against human tumor cell lines.The Journal of Antibiotics advance online publication, 15 November 2017; doi:10.1038/ja.2017.135.