Distinct promoter sequences of two precursor genes for salmon gonadotropin-releasing hormone in masu salmon

Three phase partitioning (TPP) uses t-butanol and ammonium sulfate to precipitate enzymes and proteins from aqueous solutions. The method is useful both upstream with crude samples and downstream where a scaleable simple step is needed. About 25 enzymes and proteins have been isolated by various laboratories using TPP-t-butanol. The relation of t-butanol used in TPP, with n-butanol used as an extraction agent from Morton's work, is reviewed. Some t-butanol appears bound to TPP-precipitated proteins which are actually protein-t-butanol coprecipitates. They float above denser aqueous salts because bound t-butanol increases their buoyancy, similar to the behavior of many lipoproteins. On redissolving TPP-precipitated enzymes, total and specific activities usually are regained and sometimes increased. Sulfate ion-in large concentrations-likely exerts itself through its kosmotropic action as in conventional salting out. t-Butanol likewise appears to be a kosmotrope and crowding agent at room temperature or above, whereas C1 and C2 cosolvents (e.g., ethanol) do not so behave except at near or below zero temperatures. However, kosmotropy is not the entire origin of TPP, nor probably of conventional salting out. Electrostatic forces, capacity to force protein conformation tightening and protein hydration shifts, also contribute. Electrostatic forces, and the tendency for salt ions to bind and tighten protein molecule conformation, are indicated by the sharp pH dependency of both conventional salting out and TPP, around pH regions where proteins undergo conformation changes. Sulfate anion is densely-perhaps extraordinarily-hydrated, adding much to its effective size, and therefore it has a tendency to crowd or exclude proteins, when sulfate concentrations are in the 0.5 to 3 M range.     

Effectiveness of spatial representation in the formation of network communities: experimental study on community organizer

In this paper, we present and discuss Community Organizer, a system designed to support network communities. The main characteristic of Community Organizer is the use of spatial representations for the relationships among community members including the communications exchanged among these members. These spatial representations reflect the degree of closeness of interests and concerns among the members, and are intended to provide users with clues on how to form network communities. In order to investigate the effectiveness of the proposed spatial representations, we conducted experiments with two different versions of the software. One version offered meaningful spatial representations while the other version did not. The subjects who used the former software version felt a greater sense of ‘community’, enjoyed using the software more, and actively used it longer than the subjects using the latter software version (control condition). These results indicated that the proposed spatial representations are effective in supporting network community formation.     

Influence of methylprednisolone on the intracellular antimicrobial activity of erythromycin and clindamycin against Legionella pneumophila

We have investigated the effect of methylprednisolone on the intracellular activity of erythromycin and clindamycin in vitro. An assay system was developed for the determination of intracellular activity of antibiotics against Legionella pneumophila using guinea pig resident alveolar macrophages. Erythromycin at a concentration of 0.625 mg/L (5 x MIC) and clindamycin at a concentration of 8 mg/L (MIC) inhibited the growth of a single strain of L. pneumophila in macrophages, whilst ceftizoxime at a concentration of 0.625 mg/L (5 x MIC) did not. Methylprednisolone at therapeutic concentrations did not affect the intracellular antibacterial activity of either erythromycin or clindamycin against L. pneumophila. We found no direct effect of methylprednisolone on the intracellular antibacterial activity of either erythromycin or clindamycin.     

Simplified quantitative assay system for measuring activities of drugs against intracellular Legionella pneumophila

We developed a new simple assay for the quantitation of the activities of drugs against intracellular Legionella pneumophila. The cells of a murine macrophage-like cell line (J774.1 cells) allowed the intracellular growth and replication of the bacteria, which ultimately resulted in cell death. The infected J774.1 cell monolayers in 96-well microplates were first treated with antibiotics and were further cultured for 72 h. The number of viable J774.1 cells in each well was quantified by a colorimetric assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and an enzyme-linked immunosorbent assay reader. The number of growing bacteria in each well was also determined by counting the numbers of CFU on buffered charcoal yeast extract-alpha agar plates. Viable J774.1 cell counts, determined by the colorimetric assay, were inversely proportional to the number of intracellular replicating bacteria. The minimum extracellular concentrations (MIECs) of 24 antibiotics causing inhibition of intracellular growth of L. pneumophila were determined by the colorimetric assay system. The MIECs of beta-lactams and aminoglycosides were markedly higher than the MICs in buffered yeast extract-alpha broth. The MIECs of macrolides, fluoroquinolones, rifampin, and minocycline were similar to the respective MICs. According to their intracellular activities, clarithromycin and sparfloxacin were the most potent among the macrolides or fluoroquinolones tested in this study. Our results indicated that the MTT assay system allows comparative and quantitative evaluations of the intracellular activities of antibiotics and efficient processing of a large number of samples.     

New class of cellulose fiber spun from the novel solution of cellulose by wet spinning method

A novel cellulose solution, prepared by dissolving an alkali-soluble cellulose, which was obtained by the steam explosion treatment on almost pure natural cellulose (soft wood pulp), into the aqueous sodium hydroxide solution with specific concentration (9.1 wt %) was employed for the first time to prepare a new class of multifilament-type cellulose fiber. For this purpose a wet spinning system with acid coagulation bath was applied. The mechanical properties and structural characteristics of the resulting cellulose fibers were compared with those of regenerated cellulose fibers such as viscose rayon and cuprammonium rayon commercially available. X-ray analysis shows that the new cellulose fiber is crystallographically cellulose II, and its crystallinity is higher but its crystalline orientation is slightly lower than those of other commercial regenerated fibers. The degree of breakdown of intramolecular hydrogen bond at C₃[X_am(C₃)] of the cellulose fiber, as determined by solid-state cross-polarization magic-angle sample spinning (CP/MAS) ¹³C NMR, is much lower than other, and the NMR spectra of its dry and wet state were significantly different from each other, indicating that cellulose molecules in the new cellulose fiber are quite mobile when wet. This phenomenon has not been reported for so-called regenerated cellulose fibers.     

Note: Accuracy of velocity correction for impact of a laser-accelerated miniature flyer with lithium fluoride shock-compressed along the [100] axis

We performed miniature flyer impact experiments to investigate the relationship between the apparent (u(a)) and actual (u(A)) particle velocities measured by a velocity interferometer in single-crystal lithium fluoride (LiF) that was shock-compressed along the [100] axis. The miniature flyer was accelerated to velocities in the range 652.5-1937.6 m/s by a tabletop pulsed laser. An empirical relationship of u(a) = (1.2749 ± 0.0102)u(A) was obtained. The obtained relationship agreed well with the results of a previous study within the experimental errors and its uncertainty was less than ±1%. This result indicates that the present experimental technique is effective for measuring the relationship between u(a) and u(A) of shocked transparent materials with a comparable accuracy to conventional methods.     

High-aspect-ratio through-hole array microfabricated in a PMMA plate for monodisperse emulsion production

Microchannel (MC) emulsification is a promising technique for producing monodisperse emulsions consisting of highly uniform droplets. The authors developed a high-aspect-ratio microstructure (HARMST) made of poly(methyl methacrylate) (PMMA) as a new MC emulsification device. A PMMA straight-through MC array plate consisting of 31,250 through-holes with a 7.3 × 22.9-μm oblong section and a 200-μm depth was fabricated by a process of synchrotron radiation (SR) lithography and etching. Oblong MCs fabricated in a PMMA straight-through MC array were highly uniform with a coefficient of variation of less than 2%. The fabricated PMMA straight-through MC array plate was used to produce water-in-oil (W/O) emulsions. Monodisperse W/O emulsions with average droplet diameters of approximately 25 μm and a minimum coefficient of variation of 3.2% were produced via a hydrophobic PMMA straight-through MC array. The PMMA straight-through MC array plate also produced monodisperse W/O emulsions at droplet production rates of up to 1.7 × 10⁴/s. The PMMA straight-through MC array plates developed in this work are expected to expand the application field of emulsification using straight-through MC array plates, which have previously been made of single-crystal silicon.     

Production of monodisperse water-in-oil emulsions consisting of highly uniform droplets using asymmetric straight-through microchannel arrays

This paper reports the production of monodisperse water-in-oil (W/O) emulsions using new microchannel emulsification (MCE) devices, asymmetric straight-through MC arrays that were hydrophobically modified. The silicon asymmetric straight-through MC arrays consisted of numerous pairs of microslots and circular microholes whose cross-sectional sizes were 10 μm. This paper primarily focused on investigating the effect of the osmotic pressure of a dispersed phase (Π_d) on MCE. This paper also investigated the effects of the type of continuous-phase oils and the dispersed-phase flux (J _d) on MCE. The dispersed phases were Milli-Q water and Milli-Q water solutions containing sodium chloride. The continuous phases were decane (as control), hexane, medium chain triacylglyceride (MCT), and refined soybean oil (RSO) solutions containing tetraglycerin monolaurate condensed ricinoleic acid ester (TGCR) as a surfactant. At Π_d of exceeding threshold, highly uniform aqueous droplets with coefficients of variation of less than 3% were stably generated via hydrophobic asymmetric straight-through MCs. Monodisperse W/O emulsions with average droplet diameters between 32 and 45 μm were produced using the alkane–oil and triglyceride–oil solutions as the continuous phase. This work also demonstrated that the hydrophobic asymmetric straight-through MC array had remarkable ability to produce highly uniform aqueous droplets at very high J _d of up to 1,200 L m⁻² h⁻¹.     

A low wall-loading DEMO reactor design with high priority for early and reliable realization of a tokamak fusion reactor over the cost performance

Based on scientific databases adopted for designing ITER plasmas and on the advancement of fusion nuclear technology from the recent R&D program, a low wall-loading DEMO fusion reactor has been designed, where high priority has been given to the early and reliable realization of a tokamak fusion plasma over the cost performance. Since the major radius of this DEMO reactor is chosen to be 10 m, plasma ignition is achievable with a low fusion power of 0.8 GW and an operation period of 4–5 hours is available only with inductive current drive. The low ignition power makes it possible to adopt a first wall with an austenitic stainless steel, for which significant databases and operating experience exists, due to its use in the presence of neutron irradiation in fission reactors. In step with development of advanced materials, a step-wise increase of the fusion power seems to be feasible and realistic, because this DEMO reactor has the potential to produce a fusion power of 5 GW.