Hetero-oligomerization of TDP-43 carboxy-terminal fragments with cellular proteins contributes to proteotoxicity.

Carboxy terminal fragments (CTFs) of TDP-43 contain an intrinsically disordered region (IDR) and form cytoplasmic condensates containing amyloid fibrils. Such condensates are toxic and associated with pathogenicity in amyotrophic lateral sclerosis. However, the molecular details of how the domain of TDP-43 CTFs leads to condensation and cytotoxicity remain elusive. Here, we show that truncated RNA/DNA-recognition motif (RRM) at the N-terminus of TDP-43 CTFs leads to the structural transition of the IDR, whereas the IDR itself of TDP-43 CTFs is difficult to assemble even if they are proximate intermolecularly. Hetero-oligomers of TDP-43 CTFs that have recruited other proteins are more toxic than homo-oligomers, implicating loss-of-function of the endogenous proteins by such oligomers is associated with cytotoxicity. Furthermore, such toxicity of TDP-43 CTFs was cell-nonautonomously affected in the nematodes. Therefore, misfolding and oligomeric characteristics of the truncated RRM at the N-terminus of TDP-43 CTFs define their condensation properties and toxicity.

Design and Fabrication of Mature Engineered Pre-Cardiac Tissue Utilizing 3D Bioprinting Technology and Enzymatically Crosslinking Hydrogel.

The fabrication of mature engineered cardiac tissue is one of the major challenges in cardiac tissue engineering. For this purpose, we attempted to apply the 3D bioprinting approach. Aiming to construct an oriented tissue, a fine fiber-shaped scaffold with a support structure was first designed using CAD software. Then, a 3D bioprinter and cell-adhesive bio-inks were utilized to fabricate this structure. The cell-adhesive bio-inks were synthesized by combining sodium alginate and gelatin with tyramine, respectively, to form pre-gel materials that allow enzymatic crosslinking by horseradish peroxidase. By absorbance measurements, we confirmed that the tyramine modification rate of each polymer was 0.535 mmol/g-alginate and 0.219 mmol/g-gelatin. The width of the fiber-shaped scaffold was 216.8 ± 24.3 µm for the fabricated scaffold, while the design value was 200 µm. After 3D printing and adhesion-adding treatment of the scaffold with these bio-ink materials, cardiomyocytes were seeded and cultured. As a result, the cells spread onto the scaffold, and the entire pre-tissue contracted synchronously by day 6 of culture, showing a greater pulsatility than in the early days. Video analysis showed that the beating rate of pre-myocardial tissue on day 6 was 31 beats/min. In addition, we confirmed that the cardiomyocytes partially elongated along the long axis of the fiber-shaped scaffold in the pre-tissue cultured for 15 days by staining actin, suggesting the possibility of cell orientation. Furthermore, treatment with adrenaline resulted in a 7.7-fold increase in peak beating rate compared to that before treatment (from 6 beats/min to 46 beats/min), confirming the responsiveness of the pre-tissues to the drug. These results indicate that 3D bioprinting effectively produces mature cultured myocardial tissue that is oriented, contracts synchronously, and is responsive to drugs.

Exportin-inspired artificial cell nuclear-exporting nanosystems.

Inspired by the structural and chemical features of naturally occurring importin/exportin that allows them to pass through the nuclear pore complexes, we successfully developed an artificial nuclear-exporting nanosystem capable of eliminating compounds accumulated abnormally in the nucleus.

A long-term study of the effects of SLC12A1 homozygous mutation (g.62382825G>A, p.Pro372Leu) in Japanese Black cattle.

Recessive missense mutation in the solute carrier family 12, member 1 (SLC12A1) gene (g.62382825G>A) is associated with hydrallantois, which is the accumulation of fluid in the allantoic cavity of a pregnant animal, and usually causes fetal death in Japanese Black cattle. However, the symptoms of a homozygote with this mutation that do not result in fetal death have not previously been tracked and evaluated. In the present study, we observed a homozygote with the SLC12A1 risk allele over a long-term period. The calf did not show any obvious clinical symptoms, although it did exhibit a slight growth retardation that accompanied mild calciuria. At 28 months of age, the homozygote showed renal dysfunction, which in turn resulted in hydronephrosis. The time course of the symptoms was consistent with the phenotype of Bartter syndrome in humans. Additionally, the risk heterozygous genotype did not any effects on carcass traits, which indicates that eliminating the risk allele would not have any unfavorable effects. Therefore, we emphasize that both the fetal- and late-stage symptoms associated with the SLC12A1 risk allele compromise animal welfare, and consequently may result in severe economic losses for individual farmers if the SLC12A1 risk allele is not eliminated from the population.

Weak Gravity Conjecture from Unitarity and Causality.

The weak gravity conjecture states that quantum gravity theories have to contain a charged state with a charge-to-mass ratio bigger than unity. By studying unitarity and causality constraints on higher derivative corrections to the charge-to-mass ratio of extremal back holes, we demonstrate that heavy extremal black holes can play the role of the required charged state under several assumptions. In particular, our argument is applicable when the higher-spin states Reggeizing graviton exchange are subdominant in the photon scattering. It covers (1) theories with light neutral bosons such as dilaton and moduli, and (2) UV completion where the photon and the graviton are accompanied by different sets of Regge states just like open string theory. Our result provides an existence proof of the weak gravity conjecture in a wide class of theories, including generic string theory setups with the dilaton or other moduli stabilized below the string scale.

Infinite Set of Soft Theorems in Gauge-Gravity Theories as Ward-Takahashi Identities.

We show that the soft photon, gluon, and graviton theorems can be understood as the Ward-Takahashi identities of large gauge transformation, i.e., diffeomorphism that does not fall off at spatial infinity. We found infinitely many new identities which constrain the higher order soft behavior of the gauge bosons and gravitons in scattering amplitudes of gauge and gravity theories. Diagrammatic representations of these soft theorems are presented.

Higgs inflation is still alive after the results from BICEP2.

The observed value of the Higgs boson mass indicates that the Higgs potential becomes small and flat at the scale around 10(17) GeV. Having this fact in mind, we reconsider the Higgs inflation scenario proposed by Bezrukov and Shaposhnikov. It turns out that the nonminimal coupling ξ of the Higgs squared to the Ricci scalar can be smaller than 10. For example, ξ=7 corresponds to the tensor-to-scalar ratio r≃0.2, which is consistent with the recent observation by BICEP2.