Dibenzothiophene desulfurizing enzymes from moderately thermophilic bacterium Bacillus subtilis WU-S2B: purification, characterization and overexpression

The moderately thermophilic bacterium Bacillus subtilis WU-S2B desulfurized dibenzothiophene (DBT) at 50 degrees C through the selective cleavage of carbon-sulfur bonds. In this study, three enzymes involved in the microbial DBT desulfurization were purified and characterized. The first two enzymes, DBT monooxygenase (BdsC) and DBT sulfone monooxygenase (BdsA), were purified from the wild-type strain, and the last one, 2'-hydroxybiphenyl 2-sulfinic acid desulfinase (BdsB), was purified from the recombinant Escherichia coli overexpressing the gene, bdsB, with chaperonin genes, groEL/ES. The genes of BdsC and BdsA were also overexpressed. The molecular weights of BdsC and BdsA were determined to be 200 and 174 kDa, respectively, by gel filtration chromatography, suggesting that both enzymes had four identical subunits. BdsB had a monomeric structure of 40 kDa. The three enzymes were characterized and compared with the corresponding enzymes (DszC, DszA, and DszB) of mesophilic desulfurization bacteria. The specific activities of BdsC, BdsA, and BdsB were 84.2, 855, and 280 units/mg, respectively, and the latter two activities were higher than those of DszA and DszB. The heat stability and optimum temperature of BdsC, BdsA, and BdsB were higher than those of DszC, DszA, and DszB. Other enzymatic properties were investigated in detail.     

Transmission electron microscopic study on supramolecular nanostructures of bacteriochlorophyll self-aggregates in chlorosomes of green photosynthetic bacteria

Supramolecular nanostructures of bacteriochlorophyll (BChl) self-aggregates in major light-harvesting complexes (chlorosomes) of green photosynthetic bacteria were successfully observed by freeze-fracture transmission electron microscope. Rod-shaped nanostructures with approximately 10 nm in diameter could be visualized in three types of green sulfur bacteria (Chlorobium). Diameter of the rod-shaped nanostructures in Chlorobium chlorosomes was independent of the molecular structures of their light-harvesting pigments, namely BChl-c or d. In contrast, chlorosomes of the green filamentous bacterium Chloroflexus aurantiacus had rod-shaped nanostructures with approximately 5 nm in diameter. The present results support that BChl self-aggregates in chlorosomes form rod-shaped nanostructures called rod-elements with approximately 10- and 5-nm diameters for Chlorobium and Chloroflexus, respectively.     

Analytical technique for thermoelectroelastic field in piezoelectric bodies with D∞ symmetry in cylindrical coordinates

A general solution technique for non-axisymmetric thermoelectroelastic problems in cylindrical domains with D_∞ symmetry is constructed. The displacement and electric field are expressed in terms of the respective potential functions, and the thermoelectroelastic field quantities are expressed in terms of the elastic and piezoelastic potential functions, each of which essentially satisfies a Laplace equation with respect to the spatial coordinates, combined with the two thermoelastic displacement potential functions. As an application of the technique, the theoretical analysis of a solid cylinder subjected to combined mechanical and thermal loading is performed, and the necessity of the thermoelectroelastic analyses is demonstrated.     

Cold-start performance of an ammonia-fueled spark ignition engine with an on-board fuel reformer

This work demonstrated the first-ever cold-start operation of an ammonia (NH₃)-fueled four-cylinder spark ignition engine with an on-board fuel reformer, applying autothermal reforming. In this system, an electrically heated NH₃-air mixture was provided to a reforming catalyst and approximately 3 s was found to elapse between the start of engine rotation and the onset of combustion. Stable fast idle operation in conjunction with a cold start was realized with a H₂-to-NH₃ molar ratio of 2:1. Nearly zero NH₃ emissions were achieved during cold start and fast idle until the engine warmed up, by adsorbing unburned NH₃ passing through a three-way catalyst before the catalyst was sufficiently warmed up. The NH₃ adsorption capacity of this system could be regenerated during the engine warm-up when the engine was running under lean conditions.     

Characterisation of volatile sulphur-containing compounds generated in crushed leaves of Chinese chive (Allium tuberosum Rottler)

To characterise the composition of sulphur-containing constituents in Chinese chive (Allium tuberosum Rottler), GC–MS and HPLC analyses were carried out, using 12 varieties of Chinese origin, 4 varieties of Japanese origin and 6 commercially available cultivars. Five disulphides (dimethyl, allyl methyl, methyl 1-(E)-propenyl, diallyl, and allyl 1-(E)-propenyl), two trisulphides (dimethyl and allyl methyl), and two vinyldithiins (3-vinyl-[4H]-1,2- and 2-vinyl-[4H]-1,3-) were identified on the basis of their mass spectral and GC-retention data. Among the detected sulphur compounds, sulphides with a methyl group predominated over those with an allyl group. This was supported by the HPLC estimations, suggesting that all cultivars examined had much greater amounts of methiin than alliin. Also, two novel compounds were tentatively identified as [3H,4H]-1,2-dithiin and [2H,4H]-1,3-dithiin on the basis of their mass spectral features, GC-retention data, and some experimental results on their formation in reaction mixtures with different proportions of methiin and alliin. Their formation mechanism was proposed.     

Anti‐Atherogenic Effect of Stem Cell Nanovesicles Targeting Disturbed Flow Sites

Atherosclerosis development leads to irreversible cascades, highlighting the unmet need for improved methods of early diagnosis and prevention. Disturbed flow formation is one of the earliest atherogenic events, resulting in increased endothelial permeability and subsequent monocyte recruitment. Here, a mesenchymal stem cell (MSC)‐derived nanovesicle (NV) that can target disturbed flow sites with the peptide GSPREYTSYMPH (PREY) (PMSC‐NVs) is presented which is selected through phage display screening of a hundred million peptides. The PMSC‐NVs are effectively produced from human MSCs (hMSCs) using plasmid DNA designed to functionalize the cell membrane with PREY. The potent anti‐inflammatory and pro‐endothelial recovery effects are confirmed, similar to those of hMSCs, employing mouse and porcine partial carotid artery ligation models as well as a microfluidic disturbed flow model with human carotid artery‐derived endothelial cells. This nanoscale platform is expected to contribute to the development of new theragnostic strategies for preventing the progression of atherosclerosis.     

Exfoliation of non-swelling muscovite on dodecylammonium chloride intercalation between layers using wet-jet milling

The exfoliation of layered muscovite with non-swelling property has been performed by combining various processes, such as heating, intercalation, and wet-jet milling. The c axis of muscovite was expanded from 2004.0 to 2022.8 pm at 800 °C without the destruction of crystallinity of muscovite. The heating at 800 °C led to the weak attraction force between potassium ions and silicate layers by hydroxylation of muscovite. The muscovite heated at 800 °C progressed the intercalation of dodecylammonium chloride (DDAC) into the layers effectively. Furthermore, the DDAC molecules were inserted to the interlayer of muscovite effectively by suppressing the formation of micelle of DDAC. The sedimentation test of wet-jet milled muscovite slurry showed that the relative packing density of muscovite was decreasing with increasing the amount of the intercalated DDAC. As results, the aspect ratio of muscovite prepared with combining the heating, the intercalation and the wet-jet milling was increased by 253% as compared to the raw muscovite. The aspect ratio was calculated from laser particle size distribution and thickness size distribution which was estimated from field emission-scanning electron microscopic images. The expansion of the interlayer led to the effective exfoliation of muscovite with high aspect ratio.     

Mining web access patterns with super-pattern constraint

We consider the problem of mining web access patterns with super-pattern constraint. This constraint requires that the sequential patterns in the sequence database must contain a particular set of patterns as sub-patterns. One common application of this constraint is web usage mining which mines the user access behavior on the web. In this paper, we introduce an efficient strategy for mining web access patterns with super-pattern constraint that requires only one database scan. Firstly, we present the MWAPC (M ining W eb A ccess P atterns based on super-pattern C onstraint) algorithm, in which each frequent pattern has to be checked if it contains at least one pattern from a user-defined set of patterns. Then we develop an effective algorithm, called EMWAPC that prunes the search space at the beginning of mining process and avoids checking the constraints one by one based on three proposed propositions. We have conducted the experiments on real web log databases. The experimental results show that the proposed algorithms outperform the previous methods.     

Effect of water on vapor phase photografting on cellulose and its derivatives

Effect of water on vapor phase photografting on cellulose was investigated at 60°C. An activated grafting of methyl methacrylate by water contained in the sample was observed in the experiment. The effect of water was commonly recorded irrespective of the type of cellulose derivatives such as cellulose acetate (degree of substitution, DS = 0.18 and 0.33), cellulose nitrate (DS = 0.35 and 0.75), and carboxymethl cellulose (DS = 0.19 and 0.74). Organic solvents can also be used in place of water, indicating that the percent grafting decreases in the order, water > methanol > acetone > cyclohexane. From ESR studies, water in the sample was found to contribute to the decay of cellulose radicals rather than to the radical formation. The decay was accelerated by organic solvents, and the magnitude of the effect was in the order, water ≈ methanol > acetone > cyclohexane. Based on the above investigation, it was presumed that water contained in the sample cannot contribute directly to the formation of cellulose radicals which may initiate grafting, but mostly promotes the penetration of monomer into cellulose fibers. Such penetration could lead cellulose radicals to an effective initiation of grafting.     

The mechanism of formation of wear particles of polytetrafluoroethylene

To obtain information about the basic processes involved in the formation of wear particles of polytetrafluoroethylene (PTFE), the rubbing surface of a PTFE pin was observed through a glass disk.In the initial stages of rubbing, small PTFE fragments transferred to the glass disk from the PTFE pin but did not necessarily adhere strongly to the glass disk. Some wear fragments were observed moving against the PTFE pin at a speed lower than the sliding speed of the glass disk. The fragments at higher travelling speeds combined with the fragments at lower travelling speeds and increased in size. The fragments also increased in size by scratching the surface of the PTFE pin.In steady state rubbing, large and small fragments were observed and the enlarged fragments were successively detached from the glass disk. The variation in the volume and the travelling speed of the fragments with rubbing was also examined. The total increase in the volume of transferred fragments in a definite sliding distance was found to be similar to the volume of removed fragments.