Possible activation by the green tea amino acid theanine of mammalian target of rapamycin signaling in undifferentiated neural progenitor cells in vitro

AbstractWe have shown marked promotion of both proliferation and neuronal differentiation in pluripotent P19 cells exposed to the green tea amino acid theanine, which is a good substrate for SLC38A1 responsible for glutamine transport. In this study, we evaluated the activity of the mammalian target of rapamycin (mTOR) kinase pathway, which participates in protein translation, cell growth and autophagy in a manner relevant to intracellular glutamine levels, in murine neural progenitor cells exposed to theanine. Exposure to theanine promoted the phosphorylation of mTOR and downstream proteins in neurospheres from embryonic mouse neocortex. Although stable overexpression of SLC38A1 similarly facilitated phosphorylation of mTOR-relevant proteins in undifferentiated P19 cells, theanine failed to additionally accelerate the increased phosphorylation in these stable transfectants. Theanine accelerated the formation of neurospheres from murine embryonic neocortex and adult hippocampus, along with facilitation of both 5-bromo-2’-deoxyuridine incorporation and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide reduction in embryonic neurospheres. In embryonic neurospheres previously exposed to theanine, a significant increase was seen in the number of cells immunoreactive for a neuronal marker protein after spontaneous differentiation. These results suggest that theanine activates the mTOR signaling pathway for proliferation together with accelerated neurogenesis in murine undifferentiated neural progenitor cells

In vitroNeo-Genesis of Tendon/Ligament-Like Tissue by Combination of Mohawk and a Three-Dimensional Cyclic Mechanical Stretch Culture System

Tendons and ligaments are pivotal connective tissues that tightly connect muscle and bone. In this study, we developed a novel approach to generate tendon/ligament-like tissues with a hierarchical structure, by introducing the tendon/ligament-specific transcription factor Mohawk (MKX) into the mesenchymal stem cell (MSC) line C3H10T1/2 cells, and by applying an improved three-dimensional (3D) cyclic mechanical stretch culture system. In our developed protocol, a combination of stableMkxexpression and cyclic mechanical stretch synergistically affects the structural tendon/ligament-like tissue generation and tendon related gene expression. In a histological analysis of these tendon/ligament-like tissues, an organized extracellular matrix (ECM), containing collagen type III and elastin, was observed. Moreover, we confirmed thatMkxexpression and cyclic mechanical stretch, induced the alignment of structural collagen fibril bundles that were deposited in a fibripositor-like manner during the generation of our tendon/ligament-like tissues. Our findings provide new insights for the tendon/ligament biomaterial fields

Impact of frailty on long-term mortality in older patients receiving intensive care via the emergency department

The aim of this study was to evaluate whether frailty was associated with 6-month mortality in older adults who were admitted to the intensive care unit (ICU) with an illness requiring emergency care. The investigation was a prospective, multi-center, observational study conducted among the ICUs of 17 participating hospitals. Patients >= 65 years of age who were admitted to the ICU directly from an emergency department visit were assessed to determine their baseline Clinical Frailty Scale (CFS) scores before the illness and were surveyed 6 months after admission. Among 650 patients included in the study, the median age was 79 years old, and overall mortality at 6 months was as low as 21%, ranging from 6.2% in patients with CFS 1 to 42.9% in patients with CFS >= 7. When adjusted for potential confounders, CFS score was an independent prognostic factor for mortality (one-point increase in CFS, adjusted risk ratio with 95% confidence interval 1.19 [1.09-1.30]). Quality of life 6 months after admission worsened as baseline CFS score increased. However, there was no association between total hospitalization cost and baseline CFS. CFS is an important predictor of long-term outcomes among critically ill older patients requiring emergent admission

Effect of additional magnesium on mechanical and high-cycle fatigue properties of 6061–T6 alloy

The effect of additional solute magnesium (Mg) on mechanical and high-cycle-fatigue properties of 6061-T6 aluminum alloy is investigated in detail. By adding 0.5% and 0.8% Mg to the 6061-T6 alloy with a normal stoichiometric Mg2Si composition (base alloy), the alloy exhibits eminent strain-aging characteristics demonstrated by the emergence of serrated flow, the negative strain-rate-sensitivity and relatively weakened temperature dependency of flow stress. The Mg-added new alloy also shows higher work-hardening rate than the base alloy particularly at initial flow regime and at lower strain rate. The S-N curve of the new alloy shows a clear fatigue limit which is absent in the base alloy. The fatigue limit of the new alloy is shown to be controlled by the threshold against small crack growth. Moreover, the new alloy clearly exhibits a coaxing phenomenon (time-dependent strengthening) which is absent in the base alloy. The coaxing effect is attributed to the existence of a small quasi-non-propagating crack whose growth resistance gradually increases during stress amplitude step-ups.This study was financially supported in part by the Kansai University Grant-in-Aid for progress ofresearch in graduate course (2012) and also by the Kansai University Expenditures for Support ofTraining Young Scholars (2013)

Beyond-Birthday-Bound Secure Cryptographic Permutations from Ideal Ciphers with Long Keys

Coron et al. showed a construction of a 3-round 2n-bit cryptographic permutation from three independent n-bit ideal ciphers with n-bit keys (TCC 2010). Guo and Lin showed a construction of a (2d − 1)-round dn-bit cryptographic permutation from 2d − 1 independent n-bit ideal ciphers with kn-bit keys, where d = k + 1 (Cryptography and Communications, 2015). These constructions have an indifferentiability security bound of O(q2/2n) against adversaries that make at most q queries. The bound is commonly referred to as birthday-bound security. In this paper, we show that a 5-round version of Coron et al.’s construction and (2d+1)-round version of Guo and Lin’s construction yield a cryptographic permutation with an indifferentiability security bound of O(q2/22n), i.e., by adding two more rounds, these constructions have beyond-birthday-bound security. Furthermore, under the assumption that q ≤ 2n, we show that Guo and Lin’s construction with 2d+2l−1 rounds yields a cryptographic permutation with a security bound of O(q2/2(l+1)n), where 1 ≤ l ≤ d − 1, i.e., the security bound exponentially improves by adding every two more rounds, up to 4d − 3 rounds. To the best of our knowledge, our result gives the first cryptographic permutation that is built from n-bit ideal ciphers and has a full n-bit indifferentiability security bound

Iterative Block Ciphers from Tweakable Block Ciphers with Long Tweaks

We consider a problem of constructing a secure block cipher from a tweakable block cipher (TBC) with long tweaks. Given a TBC with n-bit blocks and Γn-bit tweaks for Γ ≥ 1, one of the constructions by Minematsu in DCC 2015 shows that a simple iteration of the TBC for 3d rounds yields a block cipher with dn-bit blocks that is secure up to 2dn/2 queries, where d = Γ + 1. In this paper, we show three results.1. Iteration of 3d − 2 rounds is enough for the security up to 2dn/2 queries, i.e., the security remains the same even if we reduce the number of rounds by two.2. When the number of queries is limited to 2n, d+1 rounds are enough, and with d + l rounds for 1 ≤ l ≤ d − 1, the security bound improves as l grows.3. A d-round construction gives a block cipher secure up to 2n/2 queries, i.e., it achieves the classical birthday-bound security. Our results show that a block cipher with beyond-birthday-bound (BBB) security (with respect to n) is obtained as low as d + 1 rounds, and we draw the security spectrum of d + l round version in the range of 1 ≤ l ≤ d−1 and l = 2d−2 for BBB security, and l = 0 for birthday-bound security

Promoted neuronal differentiation after activation of alpha4/beta2 nicotinic acetylcholine receptors in undifferentiated neural progenitors.

BACKGROUND: Neural progenitor is a generic term used for undifferentiated cell populations of neural stem, neuronal progenitor and glial progenitor cells with abilities for proliferation and differentiation. We have shown functional expression of ionotropic N-methyl-D-aspartate (NMDA) and gamma-aminobutyrate type-A receptors endowed to positively and negatively regulate subsequent neuronal differentiation in undifferentiated neural progenitors, respectively. In this study, we attempted to evaluate the possible functional expression of nicotinic acetylcholine receptor (nAChR) by undifferentiated neural progenitors prepared from neocortex of embryonic rodent brains. METHODOLOGY/PRINCIPAL FINDINGS: Reverse transcription polymerase chain reaction analysis revealed mRNA expression of particular nAChR subunits in undifferentiated rat and mouse progenitors prepared before and after the culture with epidermal growth factor under floating conditions. Sustained exposure to nicotine significantly inhibited the formation of neurospheres composed of clustered proliferating cells and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction activity at a concentration range of 1 µM to 1 mM without affecting cell survival. In these rodent progenitors previously exposed to nicotine, marked promotion was invariably seen for subsequent differentiation into cells immunoreactive for a neuronal marker protein following the culture of dispersed cells under adherent conditions. Both effects of nicotine were significantly prevented by the heteromeric α4β2 nAChR subtype antagonists dihydro-β-erythroidine and 4-(5-ethoxy-3-pyridinyl)-N-methyl-(3E)-3-buten-1-amine, but not by the homomeric α7 nAChR subtype antagonist methyllycaconitine, in murine progenitors. Sustained exposure to nicotine preferentially increased the expression of Math1 among different basic helix-loop-helix proneural genes examined. In undifferentiated progenitors from embryonic mice defective of NMDA receptor subunit-1, nicotine was still effective in significantly inhibiting the proliferation. CONCLUSIONS/SIGNIFICANCE: Functional α4β2 nAChR subtype would be constitutively expressed to play a role in the mechanism underlying the determination of proliferation and subsequent differentiation fate into a neuronal lineage in association with preferential promotion of Math1 expression in undifferentiated neural progenitors of developing rodent neocortex independently of NMDA receptor activation