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This study aims to enhance the H2/N2 separation performance of polysulfone (PSF) hollow fiber membranes by annealing. X-ray diffraction, SEM, and FESEM were utilized to evaluate morphological and structural changes in denser annealed PSF membranes. AFM topography images were used to investigate the annealed form and roughness of PSF membranes. Before testing membrane permeation, it was annealed at 120, 150, and 190 °C to optimize gas separation performance. Compare to all annealing temperatures (120, 150, and 190 °C), the annealed PSF membrane at 150 °C had the greatest H2/N2 selectivity (16.18), but its separation performance was unsatisfactory since it was below the Robeson upper limit curve. The PSF before annealing and the PSF annealed at 190 °C showed outstanding separation performance (2.60 and 2.71, respectively), despite their lower selectivity than the PSF annealed at 150 °C

Purification of functional DNA nanostructures is an essential step in achieving intended functions because misfolded structures and the remaining free DNA strands in a solution can interact and affect their behavior. However, due to hydrophobicity-mediated aggregation, it is difficult to purify DNA nanostructures modified with hydrophobic molecules by conventional methods. Herein, we report the purification of cholesterol-modified DNA nanostructures using a novel surfactant-assisted gel extraction. The addition of sodium cholate (SC) to the sample solution before structure folding prevented aggregation, which was confirmed by gel electrophoresis. We also found that adding sodium dodecyl sulfate (SDS) to the sample inhibited structural folding. The cholesterol-modified DNA nanostructures prepared with SC were successfully purified by gel extraction, and their ability to bind to the lipid membrane surfaces was maintained. This method will facilitate the purification of DNA nanostructures modified with hydrophobic molecules and expand their applicability in the construction of artificial cell-like systems.

Intaking molecular information from the external environment is essential for the normal functioning of artificial cells/molecular robots. Herein, we report the design and function of a membrane nanopore using a DNA origami square tube with a cross-section of 100 nm2. When the nanopore is added to a giant vesicle that mimics a cell membrane, the permeation of large external hydrophilic fluorescent molecules is observed. Furthermore, the addition of up to four ssDNA strands enables size-based selective transport of molecules. A controllable artificial nanopore should facilitate the communication between the vesicle components and their environment.

We demonstrate a method of pattern formation based on an artificial reaction diffusion system in hydrogel medium. By designing both the reaction term and the diffusion term of the system, we have succeeded in generating a sustainable DNA pattern in the gel. A DNA logic gate anchored in the gel detected the diffused molecules from distant source points to mark the equidistant region from the source points, which produced a Voronoi pattern in the gel. Combined with a diffusion modulation method for DNA molecules, the Voronoi pattern was modified as a weighted Voronoi pattern with different morphologies. The proposed framework will be useful in designing a structured gel system responsive to molecular signals.

An isothermal amplification circuit for specific DNA molecules was implemented in giant unilamellar vesicles. Using this circuit, over 5000-fold amplification of output DNAs was achieved, and the amplification behaviour depended on the concentration of input signal DNAs in a cell-sized compartment. Moreover, initiation of the amplification by photo-stimulation was demonstrated.

The most common way to fabricate DNA nanostructures is to mix individually synthesized DNA oligomers in one pot. However, if DNA nanostructures could be produced through enzymatic reactions, they could be applied in various environments, including in vivo. Herein, an enzymatic method developed to construct a DNA nanostructure from a simple motif called a T-motif is reported. A long, repeated structure was replicated from a circular template by rolling circle amplification and then cleaved into T-motif segments by restriction enzymes. These motifs have been successfully assembled into a ladder-like nanostructure without purification or controlled annealing. This approach is widely applicable to constructing a variety of DNA nanostructures through enzymatic reactions.

In vitro transcription–translation systems (TX–TL) can synthesize most of individual genes encoded in genomes by using strong promoters and translation initiation sequences. This fact raises a possibility that TX–TL using genome as a template can reconstitute the profile of RNA and proteins in living cells. By using cell extracts and genome prepared from different organisms, here we developed a system for in vitro genome transcription–translation (iGeTT) using bacterial genome and cell extracts, and surveyed de novo synthesis of RNA and proteins. Two-dimensional electrophoresis and nano LC–MS/MS showed that proteins were actually expressed by iGeTT. Quantitation of transcription levels of 50 genes for intracellular homeostasis revealed that the levels of RNA synthesis by iGeTT are highly correlated with those in growth phase cells. Furthermore, activity of iGeTT was influenced by transcription derived from genome structure and gene location in genome. These results suggest that intracellular profiles and characters of genome can be emulated by TX–TL using genome as a template.

Cell-sized liposomes and droplets coated with lipid layers have been used as platforms for understanding live cells, constructing artificial cells, and implementing functional biomedical tools such as biosensing platforms and drug delivery systems. However, these systems are very fragile, which results from the absence of cytoskeletons in these systems. Here, we construct an artificial cytoskeleton using DNA nanostructures. The designed DNA oligomers form a Y-shaped nanostructure and connect to each other with their complementary sticky ends to form networks. To undercoat lipid membranes with this DNA network, we used cationic lipids that attract negatively charged DNA. By encapsulating the DNA into the droplets, we successfully created a DNA shell underneath the membrane. The DNA shells increased interfacial tension, elastic modulus, and shear modulus of the droplet surface, consequently stabilizing the lipid droplets. Such drastic changes in stability were detected only when the DNA shell was in the gel phase. Furthermore, we demonstrate that liposomes with the DNA gel shell are substantially tolerant against outer osmotic shock. These results clearly show the DNA gel shell is a stabilizer of the lipid membrane akin to the cytoskeleton in live cells.

A novel grinding technique is proposed with the goal of improving the grindability of the titanium alloy Ti–6Al–4V. This technique is a combination of ultrasonic-assisted grinding and plasma-electrolytic oxidation. To determine its performance in the grinding of Ti–6Al–4V, experiments were conducted to investigate the effect of ultrasonic vibration on grinding forces and work-surface roughness under the presence of plasma oxidation. The results showed that the technique drastically reduced the normal and tangential grinding forces by 60% and 70%, respectively, and decreased the work-surface roughness by 46% compared to conventional grinding, which uses neither ultrasonic assisted grinding nor plasma-electrolytic oxidation.

An empirical formula for the stress concentration factor is developed for an unequal-sized cavity pair in an arbitrary orientation. Three-dimensional finite element linear elastic analyses are performed to evaluate the stress concentration factors for different sizes, orientations, and separations of cavities. A suitable mathematical function is chosen to fit the numerical results of the finite element analyses. An application is given for evaluating the maximum stress concentration factor, which governs fatigue crack initiation in aluminum die cast test pieces from an engine block. From the X-ray CT image, the location and geometry of the gas pores are evaluated so as to develop the proposed empirical formula for this actual multi-pore system simplified to a dual spherical pore system. A proof of the formula is shown by comparison with voxel finite element analysis. The proposed empirical formula can be satisfactorily used as a scientific guideline for selecting a casting method for car engine blocks from a fatigue crack initiation perspective.

Temporal variations in the stable isotopic compositions of nitrate dissolved in stream water eluted from a cool–temperate forested watershed (8 ha) were measured to quantify the biogeochemical effects of clear-cutting of trees and subsequent strip-cutting of the understory vegetation, dwarf bamboo (Sasa senanensis), with special emphasis on changes in the fate of atmospheric nitrate that had been deposited onto the watershed based on Δ17O values of nitrate. A significant increase in stream nitrate concentration to 15 μmol L−1 in spring of 2004 was correlated with a significant increase in the Δ17O values of nitrate. Additionally, the high Δ17O values of +14.3‰ suggest that the direct drainage of atmospheric nitrate accounted for more than 50% of total nitrate exported from the forested watershed peaking in spring. Similar increases in both concentrations and Δ17O values were also found in spring of 2005. Conversely, low Δ17O values less than +1.5‰ were observed in other seasons, regardless of increases in stream nitrate concentration, indicating that the majority of nitrate exported from the forested watershed during seasons other than spring was remineralized nitrate: those retained in the forested ecosystem as either organic N or ammonium and then been converted to nitrate via microbial nitrification. When compared with the values prior to strip-cutting, the annual export of atmospheric nitrate and remineralized nitrate increased more than 16-fold and fourfold, respectively, in 2004, and more than 13-fold and fivefold, respectively, in 2005. The understory vegetation (Sasa) was particularly important to enhancing biological consumption of atmospheric nitrate., Published by Copernicus Publications on behalf of the European Geosciences Union.

Replication of all living cells relies on the multirounds flow of the central dogma. Especially, expression of DNA replication proteins is a key step to circulate the processes of the central dogma. Here we achieved the entire sequential transcription–translation–replication process by autonomous expression of chromosomal DNA replication machineries from a reconstituted transcription–translation system (PURE system). We found that low temperature is essential to express a complex protein, DNA polymerase III, in a single tube using the PURE system. Addition of the 13 genes, encoding initiator, DNA helicase, helicase loader, RNA primase and DNA polymerase III to the PURE system gave rise to a DNA replication system by a coupling manner. An artificial genetic circuit demonstrated that the DNA produced as a result of the replication is able to provide genetic information for proteins, indicating the in vitro central dogma can sequentially undergo two rounds.

Oral mucositis is one of the most common side-effects of 5-fluorouracil (5-FU)-based chemotherapy. The objective of this study was to evaluate the effects of irsogladine maleate (IM) on fluorouracil-induced oral mucositis through a double-blind, placebo controlled trial.Patients (N = 66) were randomly assigned to receive either placebo or IM (4 mg/day for 14 consecutive days). The incidence and maximum severity of fluorouracil-induced oral mucositis and safety of the irsogladine dosing regimen were evaluated.A cohort of 33 patients received placebo and 33 patients received IM. The incidence of oral mucositis was significantly lower for IM than for placebo (27% versus 73%; P0.001 by chi-square test). Specific adverse events considered related to IM were not found.IM significantly reduced the incidence and maximum severity of oral mucositis in patients treated with 5-FU-chemotherapy.

Digital cinema is a promising application that utilizes high-speed optical networks to transfer super-high-definition (SHD) images. The networks are primarily used for distributing digital cinema contents in packet data form, and are also used to support new services such as the live streaming of musicals and sport games to movie theaters. While current transfer services offer high-definition (HD) quality video, live-streaming applications will soon shift to providing cinema quality 4K content to both business and movie theaters users. The extra-high-quality 4K format enables a realistic telepresence, and will be combined with special tools such as video editing systems to realize effective remote collaboration for business workspaces. This paper introduces successive research on SHD image transmission and its application, especially in digital cinema and associated application fields.

Temperature dependence of 256 within-die random gate delay variations in sub-threshold logic circuits is measured in 40-nm CMOS test chips. When the temperature is reduced from 25 °C to -40°C, the sigma/average (σ/μ) of the gate delay at 0.3 V increases by 1.4 times. A newly developed model shows that σ/μ of the gate delay is proportional to 1/T for the first time, where T is the absolute temperature.

The 7th edition of the American Joint Committee on Cancer staging system does not include lymph node size in the guidelines for staging patients with esophageal cancer. The objectives of this study were to determine the prognostic impact of the maximum metastatic lymph node diameter (ND) on survival and to develop and validate a new staging system for patients with esophageal squamous cell cancer who were treated with definitive chemoradiotherapy (CRT).Information on 402 patients with esophageal cancer undergoing CRT at two institutions was reviewed. Univariate and multivariate analyses of data from one institution were used to assess the impact of clinical factors on survival, and recursive partitioning analysis was performed to develop the new staging classification. To assess its clinical utility, the new classification was validated using data from the second institution.By multivariate analysis, gender, T, N, and ND stages were independently and significantly associated with survival (p0.05). The resulting new staging classification was based on the T and ND. The four new stages led to good separation of survival curves in both the developmental and validation datasets (p0.05).Our results showed that lymph node size is a strong independent prognostic factor and that the new staging system, which incorporated lymph node size, provided good prognostic power, and discriminated effectively for patients with esophageal cancer undergoing CRT.

Both 3D topographic and 2D adhesive network patterns were used to guide the growth and differentiation of embryonic and mesenchymal stem cells towards a neurogenic lineage. In the case of topographic patterns, lattices composed of square micro-wells for single cell trapping combined with connection channels for neurite outgrowth guidance were designed, with optimal conditions being obtained with large well sizes and short channel lengths. In the case of adhesive micro-contact printed patterns, a similar network design can be applied, but the ''well'' size should be significantly smaller. Our results also proved the neuron-like characteristics of the induced cellular networks.

Clock skew is a major cause of severe timing yield degradation for sub-/near-threshold digital circuits. We report for the first time on employing hot-carrier injection (HCI) for post silicon clock-deskew trimming. An HCI trimmed clock buffer, which can be individually selected and stressed to adjust the clock edge, is proposed. In addition, it can be used in conjunction with on-chip skew monitoring circuits to achieve auto-stressing. Our approach is proven to be effective through a representative 1.1-mm × 0.8-mm clock tree in a 40-nm high-k complimentary metal-oxide-semiconductor process. On average, it reduces the clock skew by eight times at 0.4 V Vdd. No significant recovery is noticed two weeks after trimming.

Proteoliposomes were directly prepared by synthesizing membrane proteins with the use of minimal protein synthesis factors isolated from Escherichia coli (the PURE system) in the presence of liposomes. Connexin-43 (Cx43), which is a water-insoluble integral membrane protein that forms a hexameric complex in membranes, was cotranslationally integrated with an essentially uniform orientation in liposomes. The addition of liposomes following protein expression (post-translational presence of liposomes) did not lead to the integration of Cx43 into the liposome membranes. The amount of integrated Cx43 increased as the liposome concentration increased. The presence of liposomes did not influence the total amount of synthesized Cx43. The Cx43 integrated into the liposome membranes formed open membrane pores. These results indicate that the liposomes act in a chaperone-like manner by preventing Cx43 from aggregating in solution, because of integration into the bilayer, and also by functionalization of the integrated Cx43 in the membrane. This is the first report that cell-free-synthesized water-insoluble membrane protein is directly integrated with a uniform orientation as a functional oligomer into liposome membranes. This simple proteoliposome preparation procedure should be a valuable approach for structural and functional studies of membrane proteins. Structured digital abstract • MINT-7900670: Cx-43 (uniprotkb:P08050) and Cx-43 (uniprotkb:P08050) bind (MI:0407) by cross-linking study (MI:0030)

Objective: To evaluate the effect of maternal oral hydration on amniotic fluid index (AFI) in pregnancies with fetal gastroschisis. Methods: AFI was evaluated at 24-hour intervals, during 4 consecutive days, under a continuous maternal oral water hydration regimen, in singleton pregnancies with isolated fetal gastroschisis. Results: Nine pregnancies were examined at a mean gestational age of 31.6 weeks (±1.4) and mean maternal daily oral water intake was 3,437 (±810) ml. Mean AFI on days 0–3 were 13.2 (±2.9), 14.8 (±3.3), 14.5 (±3.1) and 14.8 (±2.6), respectively. AFI on day 0 was significantly lower compared to all the other 3 days (p = 0.01 and 0.02). Significant correlation was found in relative difference in AFI between day 0 and day 1 and gestational age (r = –0.67, p = 0.05) and the amount of water intake in the previous 24 h (r = 0.76, p = 0.02). Conclusion: Maternal oral water hydration significantly increases AFI in pregnancies with isolated fetal gastroschisis.

Purpose To identify a set of genes related to the progression and metastasis of advanced cervical cancer after radiotherapy and to establish a predictive method. Methods and Materials A total of 28 patients with cervical cancer (15 stage IIIB, 13 stage IVA patients) who underwent definitive radiotherapy between May 1995 and April 2001 were included in this study. All patients were positive for human papillomavirus infection and harbored the wild-type p53 gene. The expression profiles of 14 tumors with local failure and multiple distant metastasis and 14 tumors without metastasis (cancer free) obtained by punch biopsy were compared before treatment, using a cDNA microarray consisting of 23,040 human genes. Results Sixty-three genes were selected on the basis of a clustering analysis, and the validity of these genes was confirmed using a cross-validation test. The most accurate prediction was achieved for 63 genes (sensitivity, 78.8%; specificity, 38.1%). Some of these genes were already known to be associated with metastasis via chromosomal instability ( TTK , BUB1B ), extracellular matrix components (matrix metalloproteinase 1 [ MMP-1 ]), and carcinogenesis (protein phosphatase 1 regulatory subunit 7 [ PPP1R7 ]). A "predictive score" system was developed that could predict the probability for development of metastases using leave-one-out cross-validation methods. Conclusions The present results may provide valuable information for identified predictive markers and novel therapeutic target molecules for progression and metastasis of advanced cervical cancer.

Gardenia yellow powders A, B and C, containing geniposide at 0.284%, 0.938% and 2.783%, respectively, were administered orally to male and female SD rats as 3% feed admixtures for 13-weeks to evaluate any potential toxicity. Mean geniposide intake values were 5.72, 18.9 and 56.3 mg/kg/day in groups receiving these feed admixtures, respectively. All animals survived the duration of the study. The following findings were evident in the gardenia yellow C group: chromatouria, slightly increased plasma total bilirubin, blackish brown discoloration of the kidneys and liver, brown pigments in the proximal tubular epithelium of the kidneys. Slightly increased plasma total bilirubin was considered to be due to interference of metabolite of geniposide with the system of measurement and not to be a toxic effect since there were no related changes in histopathology of the liver or in any blood chemistry parameters. Other findings were limited to pigmentations or discolorations attributable to metabolites of geniposide. No treatment-related effects were evident on body weight, food consumption, ophthalmology, hematology or organ weights in any group. Therefore, it was concluded that 3-month ingestion of the gardenia yellow powder containing geniposide at 2.783% (approximately 60 mg/kg/day as geniposide intake) does not cause any severe toxic effects.