The flip-flop configuration of the PABP-dimer leads to switching of the translation function.

Poly(A)-binding protein (PABP) is a translation initiation factor that interacts with the poly(A) tail of mRNAs. PABP bound to poly(A) stimulates translation by interacting with the eukaryotic initiation factor 4G (eIF4G), which brings the 3' end of an mRNA close to its 5' m7G cap structure through consecutive interactions of the 3'-poly(A)-PABP-eIF4G-eIF4E-5' m7G cap. PABP is a highly abundant translation factor present in considerably larger quantities than mRNA and eIF4G in cells. However, it has not been elucidated how eIF4G, present in limited cellular concentrations, is not sequestered by mRNA-free PABP, present at high cellular concentrations, but associates with PABP complexed with the poly(A) tail of an mRNA. Here, we report that RNA-free PABPs dimerize with a head-to-head type configuration of PABP, which interferes in the interaction between PABP and eIF4G. We identified the domains of PABP responsible for PABP-PABP interaction. Poly(A) RNA was shown to convert the PABP-PABP complex into a poly(A)-PABP complex, with a head-to-tail-type configuration of PABP that facilitates the interaction between PABP and eIF4G. Lastly, we showed that the transition from the PABP dimer to the poly(A)-PABP complex is necessary for the translational activation function.

Cell-Based IL-15:IL-15Rα Secreting Vaccine as an Effective Therapy for CT26 Colon Cancer in Mice.

Interleukin (IL)-15 is an essential immune-modulator with high potential for use in cancer treatment. Natural IL-15 has a low biological potency because of its short half-life and difficulties in mass-production. IL-15Rα, a member of the IL-15 receptor complex, is famous for its high affinity to IL-15 and its ability to lengthen the half-life of IL-15. We have double-transfected IL-15 and its truncated receptor IL-15Rα into CT26 colon cancer cells to target them for intracellular assembly. The secreted IL-15:IL-15Rα complexes were confirmed in ELISA and Co-IP experiments. IL-15:IL15Rα secreting clones showed a higher anti-tumor effect than IL-15 secreting clones. Furthermore, we also evaluated the vaccine and therapeutic efficacy of the whole cancercell vaccine using mitomycin C (MMC)-treated IL-15:IL15Rα secreting CT26 clones. Three sets of experiments were evaluated; (1) therapeutics, (2) vaccination, and (3) longterm protection. Wild-type CT26-bearing mice treated with a single dose of MMC-inactivated secreted IL-15:IL-15Rα clones prolonged survival compared to the control group. Survival of MMC-inactivated IL-15:IL-15Rα clone-vaccinated mice (without any further adjuvant) exceeded up to 100%. This protection effect even lasted for at least three months after the immunization. Secreted IL-15:IL-15Rα clones challenging trigger anti-tumor response via CD4+ T, CD8+ T, and natural killer (NK) cell-dependent cytotoxicity. Our result suggested that cell-based vaccine secreting IL-15:IL-15Rα, may offer the new tools for immunotherapy to treat cancer.

Higd-1a regulates the proliferation of pancreatic cancer cells through a pERK/p27KIP1/pRB pathway.

Higd-1a/HIMP1-a/HIG1, a mitochondrial inner membrane protein, promotes cell survival under low glucose and hypoxic conditions. We previously reported that it interacts with Opa1, a factor involved in mitochondrial fusion, to regulate mitochondrial homeostasis. In the present study, we found that depletion of Higd-1a inhibited the proliferation of pancreatic cancer cells in vitro and in mice xenografts. Higd-1a knockdown did not itself lead to cell death but it caused cell cycle arrest through induction of p27KIP1 and hypo-phosphorylation of RB protein. Knockdown of Higd-1a also induced cellular senescence as shown by increased granularity and SA-ß-galactosidase activity. We further showed that the mitochondrial stress induced by Higd-1a led to reduced ERK phosphorylation. Inhibition of the ERK pathway with U0126 induced p27KIP1 expression in the pancreatic cancer cells, confirming that the cell cycle retardation was the result of inhibition of the ERK pathway. Array analysis of human pancreatic cancers revealed that expression of Higd-1a was significantly elevated in pancreatic cancer tissues compared to normal tissue. Collectively, our results demonstrate that Higd-1a plays an important role in the proliferation of pancreatic cancer cells by regulating the pERK/p27KIP1/pRB signaling pathway.

Attachment of flagellin enhances the immunostimulatory activity of a hemagglutinin-ferritin nano-cage.

Hemagglutinin (HA) displayed on a ferritin nano-cage has been shown to be effective in generating a potent immune response against a broad range of influenza infections. Here, we showed that conjugation of flagellin together with HA to the exterior surface of the ferritin cage greatly enhanced not only the humoral immune response in mice but also antigen-specific T cell responses that include Th1 cytokine secretion. The effect of flagellin remained essentially unchanged when the molar ratio of flagellin to HA was reduced from 1:1 to 1:3. Injection of the ferritin-HA-flagellin cage provided protection against lethal virus challenge in mice. We used a small immunoglobulin fragment VL12.3 as a convenient method for attaching HA and flagellin to the ferritin cage. This attachment method can be used for rapid screening of a variety of protein cages and nano-assemblies to identify the most suitable carrier and adjuvant proteins for the target antigen.

Coexistence of Low Damping and Strong Magnetoelastic Coupling in Epitaxial Spinel Ferrite Thin Films.

Low-loss magnetization dynamics and strong magnetoelastic coupling are generally mutually exclusive properties due to opposing dependencies on spin-orbit interactions. So far, the lack of low-damping, magnetostrictive ferrite films has hindered the development of power-efficient magnetoelectric and acoustic spintronic devices. Here, magnetically soft epitaxial spinel NiZnAl-ferrite thin films with an unusually low Gilbert damping parameter (<3 × 10-3 ), as well as strong magnetoelastic coupling evidenced by a giant strain-induced anisotropy field (≈1 T) and a sizable magnetostriction coefficient (≈10 ppm), are reported. This exceptional combination of low intrinsic damping and substantial magnetostriction arises from the cation chemistry of NiZnAl-ferrite. At the same time, the coherently strained film structure suppresses extrinsic damping, enables soft magnetic behavior, and generates large easy-plane magnetoelastic anisotropy. These findings provide a foundation for a new class of low-loss, magnetoelastic thin film materials that are promising for spin-mechanical devices.

Feet distance and static postural balance: implication on the role of natural stance.

The purpose of this study was to investigate 1) the effect of feet distance on static postural balance and 2) the location of natural feet distance and its possible role in the relationship of feet distance and postural balance. Static balance tests were performed on a force platform for 100 s with six different feet distances (0, 5, 10, 15, 20, 25 cm). Measures of postural balance included mean amplitude of horizontal ground reaction force (GRF) as well as the mean distance and velocity of the center of pressure (COP). All measures were discomposed into anterioposterior and mediolateral directions. ANOVA and post-hoc comparison were performed for all measures with feet distance as an independent factor. Also measured was the feet distance at the natural stance preferred by each subject. All measures significantly varied with feet distance (p<0.001). Mean distance of COP showed monotonic decrease with feet distance. Mean amplitude of horizontal GRF as well as mean velocity of COP showed U-shaped pattern (decrease followed by increase) with the minimum at the feet distance of 15 cm or 20 cm, near which the natural feet distance of 16.5 (SD 3.8) cm was located. COP is regarded to be an approximation of the center of mass (hence the resultant performance of postural control) in an inverted pendulum model with the horizontal GRF ignored. On the other hand, horizontal GRF is the direct cause of horizontal acceleration of a center of mass. The present result on horizontal GRF shows that the effort of postural control is minimized around the feet distance of natural standing and implies why the natural stance is preferred.

Four-dimensional visualization of rising microbubbles.

Four-dimensional imaging, which indicates imaging in three spatial dimensions as a function of time, provides useful evidence to investigate the interactions of rising bubbles. However, this has been largely unexplored for microbubbles, mostly due to problems associated with strong light scattering and shallow depth of field in optical imaging. Here, tracking x-ray microtomography is used to visualize rising microbubbles in four dimensions. Bubbles are tracked by moving the cell to account for their rise velocity. The sizes, shapes, time-dependent positions, and velocities of individual rising microbubbles are clearly identified, despite substantial overlaps between bubbles in the field of view. Our tracking x-ray microtomography affords opportunities for understanding bubble-bubble (or particle) interactions at microscales - important in various fields such as microfluidics, biomechanics, and floatation.