PTC124 targets genetic disorders caused by nonsense mutations

Nonsense mutations promote premature translational termination and cause anywhere from 5-70% of the individual cases of most inherited diseases. Studies on nonsense-mediated cystic fibrosis have indicated that boosting specific protein synthesis from <1% to as little as 5% of normal levels may greatly reduce the severity or eliminate the principal manifestations of disease. To address the need for a drug capable of suppressing premature termination, we identified PTC124-a new chemical entity that selectively induces ribosomal readthrough of premature but not normal termination codons. PTC124 activity, optimized using nonsense-containing reporters, promoted dystrophin production in primary muscle cells from humans and mdx mice expressing dystrophin nonsense alleles, and rescued striated muscle function in mdx mice within 2-8 weeks of drug exposure. PTC124 was well tolerated in animals at plasma exposures substantially in excess of those required for nonsense suppression. The selectivity of PTC124 for premature termination codons, its well characterized activity profile, oral bioavailability and pharmacological properties indicate that this drug may have broad clinical potential for the treatment of a large group of genetic disorders with limited or no therapeutic options.     

Optimal isotope labelling for NMR protein structure determinations

Nuclear-magnetic-resonance spectroscopy can determine the three-dimensional structure of proteins in solution. However, its potential has been limited by the difficulty of interpreting NMR spectra in the presence of broadened and overlapping resonance lines and low signal-to-noise ratios. Here we present stereo-array isotope labelling (SAIL), a technique that can overcome many of these problems by applying a complete stereospecific and regiospecific pattern of stable isotopes that is optimal with regard to the quality and information content of the resulting NMR spectra. SAIL uses exclusively chemically and enzymatically synthesized amino acids for cell-free protein expression. We demonstrate for the 17-kDa protein calmodulin and the 41-kDa maltodextrin-binding protein that SAIL offers sharpened lines, spectral simplification without loss of information, and the ability to rapidly collect the structural restraints required to solve a high-quality solution structure for proteins twice as large as commonly solved by NMR. It thus makes a large class of proteins newly accessible to detailed solution structure determination.     

Control of excessive neural circuit excitability and prevention of epileptic seizures by endocannabinoid signaling

Progress in research on endocannabinoid signaling has greatly advanced our understanding of how it controls neural circuit excitability in health and disease. In general, endocannabinoid signaling at excitatory synapses suppresses seizures by inhibiting glutamate release. In contrast, endocannabinoid signaling promotes seizures by inhibiting GABA release at inhibitory synapses. The physiological distribution of endocannabinoid signaling molecules becomes disrupted with the development of epileptic focus in patients with mesial temporal lobe epilepsy and in animal models of experimentally induced epilepsy. Augmentation of endocannabinoid signaling can promote the development of epileptic focus at initial stages. However, at later stages, increased endocannabinoid signaling delays it and suppresses spontaneous seizures. Thus, the regulation of endocannabinoid signaling at specific synapses that cause hyperexcitability during particular stages of disease development may be effective for treating epilepsy and epileptogenesis.     

基于建筑类型学的南京中华门门西地区传统民居平面空间演变

以中国古都"城市组织"为研究背景,以古都南京中华门门西历史街区为考察对象,用"建筑类型学"的分析方法对传统民居进行分析,旨在为今后传统民居的保护、传承和利用提供依据.     

Understanding paternal genome demethylation through live-cell imaging and siRNA

Identification of a DNA demethylase responsible for zygotic paternal DNA demethylation has been one of the most challenging goals in the field of epigenetics. Several candidate molecules have been proposed, but their involvement in the demethylation remains controversial, partly due to the difficulty of preparing a sufficient quantity of materials for biochemical analysis. In this review, we utilize a recently developed method for live-cell imaging of mouse zygotes combined with RNA interference (RNAi) to search for factors that affect zygotic paternal DNA demethylation. The combined use of various fluorescent probes and RNAi is a useful approach for the study of not only DNA demethylation but also the spatiotemporal dynamics of histone depositions in zygotes, although it is not appropriate for large-scale screening or knockdown of genes that are abundantly expressed before fertilization. This new technique enables us to understand the epigenetic hierarchy during cellular response and differentiation in preimplantation embryos.     

Implementation of 3-Valued Paraconsistent Logic Programming Towards Decision Making System of Agents

Due to the rapid development of applications of artificial intelligence and robotics in recent years, the necessity of reasoning and decision making with uncertain and inaccurate information is increasing. Since robots in the real world are always exposed to behavioral inaccuracies and uncertainty arising from recognition methods, they may occasionally encounter contradictory facts during reasoning on action decision.Paraconsistent logic programming is promising to make appropriate action decisions even when an agent is exposed to such uncertain information or contradictory facts, but there has been no implementation of this programming to the best of our knowledge. We propose a resolution algorithm for the 3-valued paraconsistent logic programming system QMPT0 and its implementation on SWI-Prolog. We also describe an application of the 3-valued paraconsistent logic programming regarding agent decision making.     

Periodicity in fish otolith Sr,Na, and K corresponds with visual banding

Examination of a section of an otolith from a teleost fish by fast atom bombardment-secondary ion mass spectrometry (FAB-SIMS) revealed seasonal periodicity in Sr, Na, and K concentrations that corresponded with visually observed annual banding. Strontium maxima also corresponded with Na and K minima and vice versa. In addition there was a general, apparently age-related, trend in Sr levels, with concentrations at the edge of the otolith being higher than in the core.     

Current cases in which epitope mimicry is considered a component cause of autoimmune disease: Guillain-Barré syndrome

Some patients develop Guillain-Barré syndrome (GBS) after the administration of bovine gangliosides. Patients with GBS subsequent to Campylobacter jejuni enteritis frequently have IgG antibody to GM1 ganglioside. Miller Fisher syndrome (MFS), a variant of GBS, is associated with IgG antibody to GQ1b ganglioside. Molecular mimicry between GM1 and lipopolysaccharide of C. jejuni isolated from patients with GBS, and between GQ1b and C. jejuni lipopolysaccharides from patients with MFS have been demonstrated. The molecular mimicry between infectious agents and gangliosides may function in the production of anti-ganglioside antibodies. This sugar mimicry is one possible cause of the Guillain-Barré and Miller Fisher syndromes; however, unidentified host factors may contribute to the development of these syndromes.     

Measurement of blood flow by heated thermocouple with feedback controlled current

Zusammenfassung Eine neue Messmethode für Durchblutungsmessungen besteht darin, dass selbstgeregelter Strom mit Hilfe des Rückkopplungsregelungssystems für die Heizung gebraucht wird. Mit dieser Methode wird der Temperaturunterschied zwischen den geheizten und den gekühlten Kontaktplatten ganz gering und konstant gehalten, so dass der Effekt der lokalen Wärmeakkumulation, der bei Heizung mit konstantem Strom entsteht, am geprüften Gewebe wegfällt. Diese Methode hat eine höhere Linearität und bessere Anwendungsfähigkeit über weitere Durchblutungsgebiete, und sie spricht auch schneller an als die frühere Methode der Heizung mit konstantem Strom.