Integrated molecule-format bioluminescent probe for visualizing androgenicity of ligands based on the intramolecular association of androgen receptor with its recognition Peptide

Ligand-induced conformational changes of nuclear hormone receptors (NRs) are an important start-up of various hormone signaling. However, little is known of the bioanalytical use of the hormone-induced conformational changes of NRs. Here, we describe a generally applicable bioluminescence assay with a genetically encoded bioluminescent indicator to determine androgenicity of ligands based on the intramolecular association of the ligand binding domain of androgen receptor (AR LBD) with the "FQNLF" motif in the N-terminal domain of AR (AR NTD). Firefly luciferase (FLuc) was dissected into N-terminal (1-415 AA) and C-terminal (416-550 AA) fragments. The AR LBD and FQNLF motif of AR NTD were sandwiched between the dissected fragments of FLuc to construct a single molecule-format bioluminescent probe. Androgens induce the association of AR LBD with the FQNLF motif in the NTD, and the subsequent complementation of N- and C-terminal fragments of FLuc partially restores the activities of FLuc. A 10-5 M solution of 5alpha-dihydroxytestosterone (DHT) induced a quick increase in the luminescence intensities from cervical carcinoma-derived HeLa cells carrying the genetic indicator, which reached a plateau in 9 min, whereas DHT withdrawal from the cells by a medium change decreased the luminescence with a slower time course, i.e., approximately 2 h until returning to the background luminescence. The present luminescent indicator was found to exhibit high agonist selectivity and reproducible recovery of the luminescence to a repeated androgen addition and withdrawal. This is the first contribution that cellular signaling steps can be imaged with bioluminescence using a single molecule-format bioluminescence probe (Simbi), in which all the components required for a signal sensing and visualization are integrated. Simbi is applicable to developing biotherapeutic agents effecting to the AR signaling, and for screening adverse chemicals that possibly influence the signal transduction of AR.     

Air‐Stable n‐Type Organic Field‐Effect Transistors Based on Carbonyl‐Bridged Bithiazole Derivatives

An electronegative conjugated compound composed of a newly designed carbonyl‐bridged bithiazole unit and trifluoroacetyl terminal groups is synthesized as a candidate for air‐stable n‐type organic field‐effect transistor (OFET) materials. Cyclic voltammetry measurements reveal that carbonyl‐bridging contributes both to lowering the lowest unoccupied molecular orbital energy level and to stabilizing the anionic species. X‐ray crystallographic analysis of the compound shows a planar molecular geometry and a dense molecular packing, which is advantageous to electron transport. Through these appropriate electrochemical properties and structures for n‐type semiconductor materials, OFET devices based on this compound show electron mobilities as high as 0.06 cm² V^?1 s^?1 with on/off ratios of 10⁶ and threshold voltages of 20 V under vacuum conditions. Furthermore, these devices show the same order of electron mobility under ambient conditions.     

Ultra-high-speed deformation by impact of a projectile flying at speeds on the order of km s

An apparatus was developed to facilitate application of an electro-thermo-chemical accelerator to high-speed deformation experiments. The apparatus is designed on the principle of sequential collision of elastic bodies. Speeds ranging from 600 to 780 m s⁻¹ were achieved, and estimated strain rate of deformation is 10⁷ s⁻¹. The newly developed apparatus can be applied to various types of accelerators for attaining deformation speeds as high as several km s⁻¹. Transmission electron microscopy of aluminum deformed at high speed by use of the apparatus revealed the formation of very small stacking fault tetrahedra (SFTs). This observation is quite new for aluminum; previously, SFTs had not been observed in aluminum, although deformation had been carried out at strain rates lower than 10⁶ s⁻¹. Use of the apparatus promises to provide new insight into high-speed deformation.     

Findings of MR and MR cholangiopancreatography in acute cholecystitis

We retrospectively reviewed magnetic resonance cholangiopancreatography (MRCP) of 25 patients with acute cholecystitis based on clinical, sonographic and surgical findings. Intramural high signal intensity on MRCP was demonstrated in 22 of the 25 patients (88%), and pericholedochal high signal intensity was observed in 6 of the 25 patients (24%). Pericholecystic or perihepatic fluid was demonstrated in 6 of the 25 patients (24%). Gallbladder stones were identified in all 21 patients (100%) by sonography, in 19 of the 21 (90%) by MRCP and in 11 of 18 patients by CT (CT was not performed in other 3 patients). Common bile duct calculi were detected in all 6 patients (100%) by MRCP, in 2 of the 6 (33%) by sonography, and in 5 of the 6 (50%) by CT with confirmation of surgical finding or endoscopic retrograde cholangiography (ERC). MRCP had a high accuracy in diagnosing acute cholecystitis with the finding of intramural high signal intensity. MRCP is an excellent method to evaluate acute biliary disease and may replace CT and ERC in the preoperative evaluation of acute cholecystitis.     

Quinine Actinometry as a Method for Calibrating Ultraviolet Radiation Intensity in Light-Stability Testing of Pharmaceuticals

A collaborative study was carried out by seven different laboratories to evaluate quinine actinometry as a universal, standardized method for calibrating UV radiation intensity from light sources used in light-stability testing of pharmaceutical products. Near UV fluorescent lamps, white fluorescent lamps, metal halide lamps and xenon arc lamps were employed as light sources. The increase in absorbance at 400 nm of aqueous quinine solutions was found to be proportional to the integrated UV energy emitted from the light sources. The linearity observed between absorbance and integrated UV energy indictates that quinine actinometry can be used to measure the intensity of UV radiation at wavelengths around 330 nm. The slopes of regression curves of absorbance vs integrated UV energy varied among the lamps used due to differing spectral distributions. Light degradation of nifedipine, a model photosensitive drug, was studied based on quinine actinometry.     

The mechanism of myometrial contractions induced by endothelin-1 in rat

Experiments were performed to characterize endothelin-1-induced contractions and the role of endothelin (ET) receptor subtypes in rat myometrium. The binding sites of [(125)I]-ET-1 were saturable with high affinity. Scatchard plot analysis revealed that ET-1 binding sites in the myometrium constituted a single population. The dissociation equilibrium constant (Kd) and the maximum binding sites (Bmax) were determined to be 48.9+/-3.0 pM and 1364.0+/-210.3 fmol/mg protein respectively. Specific [(125)I]-ET-1 binding was inhibited completely by unlabelled ET-1 and Ro 46-2005 (mixed-type ET receptor antagonist), but not fully (90.7+/-1.4%) by BQ 123 (a selective ETA receptor antagonist), and not at all by RES 701-1 (a selective ETB receptor antagonist). ET-1 induced myometrial contractions were composed of two types, an increase in resting tone and rhythmic contractions. These contractions were inhibited by BQ 123 and Ro 46-2005, but not by RES 701-1. ET-1-induced contractions were greatly reduced in Ca2+-free Krebs' solution. Nifedipine abolished the rhythmic contractions without affecting the increase in resting tone. These results suggest that ETA receptors are predominantly localized in rat myometrium and that excitation of ETA receptors evokes two types of contractions by increasing the cytoplasmic Ca2+ concentration.     

Dynamical characteristics of linear cellular automata

Dynamical characteristics of linear cellular automata are discussed algebraically, whose cell space and state space are an Abelian group and a finite commutative ring, respectively, instead of a lattice space and a residue class. One of the main results is a characterization of the dynamical structures with relation to what the unit configuration is. It is also shown that a linear cellular automaton with the state space of a residue class of an integer m can be decomposed in parallel into automata with the one of a power of a prime which is a factor of m. Using those results, the proofs of known results are improved concerning C-surjectivity, C-injectivity, and finite-order property for linear cellular automata and presented in a unified manner.     

Evolutionary design of high signal integrity interconnection based on eye-diagram

Frequency of the digital signal in PCBs has remained at about 1 GHz, while that in VLSIs become more than several GHz. The frequency gap comes from signal integrity (SI) deterioration in PCB in GHz-domain. To overcome this problem, we have already proposed segmental transmission line, in which genetic algorithm is used to solve the combinatorial explosion problem in the design. In this paper, we propose a novel fitness based on the eye-diagram, which is widely and generally used for SI evaluation in the high-speed interconnection design field. And we also propose a high-speed eye-diagram calculation method based on virtual eye-diagram. This eye-diagram is constructed by superposing the single-shot pulse responses and can reduce the simulation time. The simulation experiments demonstrate that our proposed fitness increases the eye height and eye width by 3.07 and 1.06 times, respectively, compared with the conventional one.     

Pattern recognition using boundary data of component distributions

In statistical pattern recognition, a Gaussian mixture model is sometimes used for representing the distribution of vectors. The parameters of the Gaussian mixture model are usually estimated from given sample data by the expectation maximization algorithm. However, when the number of data attributes is large, the parameters cannot be estimated correctly. In this paper, we propose a novel approach for estimating the parameters of the Gaussian mixture model by using sample data located on the boundary of regions defined by the component density functions. Experiments are carried out to show the characteristics of the proposed method.     

Development of a novel crossover of hybrid genetic algorithms for large-scale traveling salesman problems

This article proposes a novel crossover operator of hybrid genetic algorithms (HGAs) with a Lin-Kernighan (LK) heuristic for solving large-scale traveling salesman problems (TSPs). The proposed crossover, tentatively named sub-tour recombination crossover (SRX), collects many short sub-tours from both parents under some set of rules, and reconnects them to construct a new tour of the TSP. The method is evaluated from the viewpoint of tour quality and CPU time for ten well-known benchmarks, e.g., dj38, qa194, …, ch71009.tsp, in the TSP website of the Georgia Institute of Technology. We compare the SRX with three conventional crossover operators, a variant of the maximal preservative crossover operator (MPX3), a variant of the greedy sub-tour crossover operator (GSX2), and a variant of the edge recombination crossover operator (ERX6), and show that the SRX succeeded in finding a better solution and running faster than the conventional methods mentioned above.