Structural basis of the initial binding of tRNA(Ile) lysidine synthetase TilS with ATP and L-lysine

In the bacterial genetic-code system, the codon AUA is decoded as isoleucine by tRNA(Ile)(2) with the lysidine residue at the wobble position. Lysidine is derived from cytidine, with ATP and L-lysine, by tRNA(Ile) lysidine synthetase (TilS), which is an N-type ATP pyrophosphatase. In this study, we determined the crystal structure of Aquifex aeolicus TilS, complexed with ATP, Mg2+, and L-lysine, at 2.5 A resolution. The presence of the TilS-specific subdomain causes the active site to have two separate gateways, a large hole and a narrow tunnel on the opposite side. ATP is bound inside the hole, and L-lysine is bound at the entrance of the tunnel. The conserved Asp36 in the PP-motif coordinates Mg2+. In these initial binding modes, the ATP, Mg2+, and L-lysine are held far apart from each other, but they seem to be brought together for the reaction upon cytidine binding, with putative structural changes of the complex.     

Flexible manipulation of microfluids using optically regulated adsorption/desorption of hydrophobic materials

To realize highly integrated micro total analysis systems (microTAS), a simply controlled miniaturized valve should be utilized on microfluidic device. In this paper, we describe the application of photo-induced super-hydrophilicity of titanium dioxide (TiO2) to microfluidic manipulation. In addition, we found a new phenomenon for reversibly converting the surface wettability using a polydimethylsiloxane (PDMS) matrix and the photocatalytic properties of TiO2. While PDMS polymer was irradiated with UV, it was confirmed that hydrophobic material was released from the polymer to air. Several prepolymers were identified as the hydrophobic material with a gas chromatograph and mass spectrometer (GC/MS). Here, we successfully demonstrated the flexible manipulation of microfluid in a branched microchannel using the reversible wettability as micro opto-switching valve (MOS/V). The simultaneous control of MOS/Vs was also demonstrated on a 256-MOS/V integrated disk. The MOS/V promises to be one of the most effective flow switching valves for advanced applications in highly integrated micro/nano fluidics.     

Feasibility study of a pilot-scale sewage treatment system combining an up-flow anaerobic sludge blanket (UASB) and an aerated fixed bed (AFB) reactor at ambient temperature

A feasibility test of a 17 m3-pilot-scale sewage treatment system was carried out by continuous feeding of raw municipal sewage under ambient temperature conditions. The system consisted of a UASB and an aerated fixed bed reactor. Some of the effluent from the fixed bed reactor was returned to the UASB influent in order to provide a sulfate source. The total BOD of 148-162 mg l(-1) in the influent was reduced to a more desirable 11-25 mg l(-1) in the final effluent. The levels of methane-producing activity from acetate and H2/CO2 gas at 10 degrees C were only 2% and 0% of those at 35 degrees C, respectively. On the other hand, the sulfate-reducing activity levels of the UASB sludge were relatively high at 10 degrees C, for example, 18% for acetate and 9% for H2/CO2 gas, compared to the activity levels at 35 degrees C. Therefore, BOD oxidization by sulfate reduction in the UASB was greater than that by methane production under low temperature conditions. This sulfate-reducing activity tended to be proportional to the copy number of adenosine-5'-phosphosulfate (APS) reductase genes in DNA extracted from the sludge.     

Synthesis of fluorescent-labeled aeruginosin derivatives for high-throughput fluorescence correlation spectroscopy assays

The design and solid-phase synthesis of effective fluorescent-labeled aeruginosin derivatives and their application to the fluorescence correlation spectroscopy (FCS)-based competitive binding assay of an aeruginosin library are described. The phenolic hydroxyl group on the (R)-3-(4-hydroxyphenyl)lactic acid (d-Hpla) residue was observed to be suitable for connecting Rhodamine green derivative with minimum loss of biological activity. In addition, the FCS-based binding assay of the library using fluorescent-labeled chemical probes was also achieved.     

Design, synthesis, and biological activity of folate receptor-targeted prodrugs of thiolate histone deacetylase inhibitors

Aiming to develop selective anticancer drugs, we designed and synthesized three disulfides bearing a folic acid moiety as candidate folate receptor (FR)-targeted prodrugs of thiolate histone deacetylase inhibitors. Among them, compound 1 displayed growth-inhibitory activity toward folate receptor-positive MCF-7 breast cancer cells. The activity of 1 was significantly reduced by free folic acid, suggesting that cellular uptake of 1 is mediated by FR.     

Arabidopsis KAM2/GRV2 is required for proper endosome formation and functions in vacuolar sorting and determination of the embryo growth axis

We isolated an Arabidopsis thaliana mutant, katamari2 (kam2), that has a defect in the organization of endomembranes. This mutant had deformed endosomes and formed abnormally large aggregates with various organelles. Map-based cloning revealed that kam2 is allelic to gravitropism defective 2 (grv2). The KAM2/GRV2 gene encodes a homolog of a DnaJ domain-containing RECEPTOR-MEDIATED ENDOCYTOSIS-8, which is considered to play a vital role in the endocytotic pathway from the plasma membrane to lysosomes in animal cells. Immunofluorescent staining showed that KAM2/GRV2 protein localizes on punctate structures, which did not merge with any markers for Golgi, trans-Golgi network, endosomes, or prevacuolar compartments. KAM2/GRV2, which does not have a predicted transmembrane domain, was peripherally associated with the membrane surface of uncharacterized compartments. KAM2/GRV2 was expressed at the early to middle stages of seed maturation. We found kam2 mis-sorted seed storage proteins by secreting them from cells, indicating that KAM2/GRV2 is involved in the transport of the proteins into protein storage vacuoles. kam2 had another defect in embryogenesis. Half of the developing kam2-1 cotyledons grew into the opposite space of the seeds before the walking stick-shaped embryo stage. Our findings suggest that KAM2/GRV2 is required for proper formation of the endosomes involving protein trafficking to the vacuoles and determination of growth axis of the embryo.     

The development of trunk muscles in male wrestlers assessed by magnetic resonance imaging

The purpose of this study was to compare the development of trunk musculature among Elite, Sub-elite, and Elite junior wrestlers. The performance level of these groups, ordered highest to lowest, is as follows: Elite (n = 20), Sub-elite (n = 25), and Elite junior (n = 39). A magnetic resonance imaging device was used to measure the cross-sectional area (CSA) of the trunk muscles. The whole trunk muscle cross-sectional area (t-MCSA) of the Elite group was significantly larger than that of the Elite junior group (p < 0.05). The rectus abdominis muscle CSA of the Elite group was significantly larger than that of the Elite junior group (p < 0.01). The psoas major muscle CSA of the Elite group was significantly larger than that of the Elite junior group (p < 0.05). There were no significant differences in the CSA of any of the trunk muscles between the Elite and Sub-elite groups. In conclusion, compared with Elite junior wrestlers, it is conceivable that a greater CSA of trunk flexors of Elite wrestlers is one factor which supports increased performance.     

Intranasal immunization with H5N1 vaccine plus Poly I:Poly C12U, a Toll-like receptor agonist, protects mice against homologous and heterologous virus challenge

The avian H5N1 influenza virus has the potential to cause a new pandemic. Since it is difficult to predict which strain of influenza will cause a pandemic, it is advantageous to produce vaccines that confer cross-protective immunity. Mucosal vaccine administration was reported to induce cross-protective immunity by inducing secretion of IgA at the mucosal surface. Adjuvants can also enhance the development of fully protective mucosal immunity. Here we show that a new mucosal adjuvant, poly I:poly C12U (Ampligen), a Toll-like receptor 3 agonist proven to be safe in a Phase III human trial, is an effective adjuvant for H5N1 influenza vaccination. Intranasal administration of a candidate influenza vaccine with Ampligen resulted in secretion of IgA, and protected mice that were subsequently challenged with homologous A/Vietnam/1194/2004 and heterologous A/HK/483/97 and A/Indonesia/6/2005 virus.