Mechanism‐Based Trapping of the Quinonoid Intermediate by Using the K276R Mutant of PLP‐Dependent 3‐Aminobenzoate Synthase PctV in the Biosynthesis of Pactamycin

Mutational analysis of the pyridoxal 5′‐phosphate (PLP)‐dependent enzyme PctV was carried out to elucidate the multi‐step reaction mechanism for the formation of 3‐aminobenzoate (3‐ABA) from 3‐dehydroshikimate (3‐DSA). Introduction of mutation K276R led to the accumulation of a quinonoid intermediate with an absorption maximum at 580 nm after the reaction of pyridoxamine 5′‐phosphate (PMP) with 3‐DSA. The chemical structure of this intermediate was supported by X‐ray crystallographic analysis of the complex formed between the K276R mutant and the quinonoid intermediate. These results clearly show that a quinonoid intermediate is involved in the formation of 3‐ABA. They also indicate that Lys276 (in the active site of PctV) plays multiple roles, including acid/base catalysis during the dehydration reaction of the quinonoid intermediate.     

Nanoparticle formation in H2O/N2 and H2O/Ar mixtures under irradiation by 20 MeV protons and positive corona discharge

To investigate the contribution of ions to gas nucleation, we have performed experiments on the formation of water droplets in H₂O/N₂ and H₂O/Ar gas mixtures by irradiation with a 20 MeV proton beam and by positive corona discharge. The size of the formed nanoparticles was measured using a differential mobility analyzer equipped with a Faraday cup electrometer. Using the proton beam, droplets around 10 nm in diameter were observed for both positively and negatively charged particles, but none were found when the corona discharge was used. This implies the importance of the presence of both positive and negative ions for the formation of nanosize droplets, which attract each other by Coulomb interactions, enhancing the collision frequency and leading to the formation of the 10 nm droplets.     

Homogeneous and Disordered Assembly of Densely Packed Nanocrystals

A homogeneous and disordered assembly of densely packed nanocrystals is generated without the assistance of organic molecules, from an aqueous solution at room temperature. The densely packed nanocrystals of tin dioxide (SnO₂) and titanium dioxide (TiO₂), 2–3 nm in size, form glassy macroscopic transparent objects of 2–10 mm. The monodisperse nanocrystals, which have a surface hydrated layer and are synthesized in aqueous solution, are densely packed in an homogeneous and disordered assembly through the evaporation of water as the dispersion medium. The formation of transparent bulk objects on the macroscopic scale originates from the homogeneous and disordered nature of the densely packed nanocrystals. The resultant homogeneous architectures are distinguishable from the typical inhomogeneous and disordered aggregates of nanoparticles observed after evaporation of the dispersion medium. Among superstructures based on nanocrystals, such as superlattices and mesocrystals, the resultant homogeneous and disordered assembly can be regarded as a new class of solid‐state material.     

Cloning and Characterization of the Biosynthetic Gene Cluster of 16‐Membered Macrolide Antibiotic FD‐891: Involvement of a Dual Functional Cytochrome P450 Monooxygenase Catalyzing Epoxidation and Hydroxylation

FD‐891 is a 16‐membered cytotoxic antibiotic macrolide that is especially active against human leukemia such as HL‐60 and Jurkat cells. We identified the FD‐891 biosynthetic (gfs) gene cluster from the producer Streptomyces graminofaciens A‐8890 by using typical modular type I polyketide synthase (PKS) genes as probes. The gfs gene cluster contained five typical modular type I PKS genes (gfsA, B, C, D, and E), a cytochrome P450 gene (gfsF), a methyltransferase gene (gfsG), and a regulator gene (gfsR). The gene organization of PKSs agreed well with the basic polyketide skeleton of FD‐891 including the oxidation states and α‐alkyl substituent determined by the substrate specificities of the acyltransferase (AT) domains. To clarify the involvement of the gfs genes in the FD‐891 biosynthesis, the P450 gfsF gene was inactivated; this resulted in the loss of FD‐891 production. Instead, the gfsF gene‐disrupted mutant accumulated a novel FD‐891 analogue 25‐O‐methyl‐FD‐892, which lacked the epoxide and the hydroxyl group of FD‐891. Furthermore, the recombinant GfsF enzyme coexpressed with putidaredoxin and putidaredoxin reductase converted 25‐O‐methyl‐FD‐892 into FD‐891. In the course of the GfsF reaction, 10‐deoxy‐FD‐891 was isolated as an enzymatic reaction intermediate, which was also converted into FD‐891 by GfsF. Therefore, it was clearly found that the cytochrome P450 GfsF catalyzes epoxidation and hydroxylation in a stepwise manner in the FD‐891 biosynthesis. These results clearly confirmed that the identified gfs genes are responsible for the biosynthesis of FD‐891 in S. graminofaciens.     

Ethylamine content of fresh tea shoots and made tea determined by high performance liquid chromatography

The ethylamine contents of tea shoots and made teas were derived with o-phthalaldehyde and 2-mercaptoethanol, and quantified by high performance liquid chromatography. The coefficient of variation of this method was 1.97% from eight analyses. The levels of ethylamine in tea shoots declined markedly during growth and senescence after plucking; they also declined in the extracts of fresh tea during the incubation at 37°C, but this decrease was completely inhibited by amine oxidase inhibitors. The content of ethylamine in high grade green teas was higher than in low grade teas, but was almost identical in high and low grade black tea. The mean contents of ethylamine in 22 green teas and 17 black teas were 6.76±3.07 μmol g^?1 and 6.40±1.82 μmol g^?1, respectively.     

Gaussian process motion graph models for smooth transitions among multiple actions

We propose a unified model for human motion prior with multiple actions. Our model is generated from sample pose sequences of the multiple actions, each of which is recorded from real human motion. The sample sequences are connected to each other by synthesizing a variety of possible transitions among the different actions. For kinematically-realistic transitions, our model integrates nonlinear probabilistic latent modeling of the samples and interpolation-based synthesis of the transition paths. While naive interpolation makes unexpected poses, our model rejects them (1) by searching for smooth and short transition paths by employing the good properties of the observation and latent spaces and (2) by avoiding using samples that unexpectedly synthesize the nonsmooth interpolation. The effectiveness of the model is demonstrated with real data and its application to human pose tracking.     

An experimental study for transmitting MPEG-2 streams over ATM networks

This paper presents a comparative study of different techniques aiming at transmitting MPEG-2 streams over ATM networks, with emphasis on the presentation of practical results achieved on an experimental platform available at LSI, a laboratory of the University of São Paulo, Brazil. This study has the purpose to determine some tuning parameters for optimizing the implementation of distributed multimedia applications on different network technologies. The focus of this experimental study has been on non-native ATM techniques like “Classical IP over ATM” and “LAN Emulation”, as well as native ATM techniques based on direct access service primitives to AAL5. The influence of different parameters, like packet length, buffer size, CPU speed, on the resulting throughput has been studied. Furthermore, a testing tool is being developed for assessing the transmission of MPEG-2 streams based on the practical experience achieved so far. These results will permit the evaluation of different network technologies and also help in the development of distributed multimedia applications like video-on-demand, videoconferencing and telemedicine.     

Pb-free surface-finishing on electronic components’ terminals for Pb-free soldering assembly

Pb-free solderable surface finishing is essential to implement Pb-free solder assembly in order to meet with the growing demand of environmental consciousness to eliminate Pb from electronic products. Two types of widely applicable Pb-free surface finishing technologies are developed. One is the multilayer-system including Pd with Ni undercoat. Heat-resistance of Pd enables whole-surface-plating on to leadframe before IC-assembling process. The other is the double-layer-system with low-melting-point-materials, for example, thicker Sn underlayer and thinner Sn-Bi alloy overlayer, dilutes Sn-Bi alloy’s defects of harmful reactivity along with substrate metal and mechanical brittleness with keeping its advantages of solder-wettability and no whisker.     

Temporal segmentation and assignment of successive actions in a long-term video

Temporal segmentation of successive actions in a long-term video sequence has been a long-standing problem in computer vision. In this paper, we exploit a novel learning-based framework. Given a video sequence, only a few characteristic frames are selected by the proposed selection algorithm, and then the likelihood to trained models is calculated in a pair-wise way, and finally segmentation is obtained as the optimal model sequence to realize the maximum likelihood. The average accuracy on IXMAS dataset reached to 80.5% at frame level, using only 16.5% of all frames in computation time of 1.57 s per video which has 1160 frames on the average.