Development of a novel method for operating magnetic particles, Magtration Technology, and its use for automating nucleic acid purification

Magnetic particles are useful for simple and efficient nucleic acid extraction. To achieve fully automated nucleic acid extraction and purification using magnetic particles, a new method for operating magnetic particles, Magtration Technology, was developed. In this method, magnetic separation is performed in a specially designed disposable tip. This enables high recovery of magnetic particles with high reproducibility. The features of this technology are (i) a simple mechanism for process control and (ii) flexible software to enable adaptation to commercially available reagents. Automated instruments based on Magtration Technology were developed and used for nucleic acid extraction. Total DNA, total RNA and plasmids were purified by Magtration Technology at an efficiency comparable to that of manual methods.     

Drying process in a solvent-based paint analyzed by phase-shifting digital holography and an estimation of time for tack free

A technique to study the drying of paints, based on phase-shifting digital holography, is presented. The technique is applied to the drying process of solvent-based paint on a three-dimensional surface at different substrate temperatures. For processing the data, a cross-correlation function and phase change derived from reconstructed complex amplitudes are calculated to visualize and to evaluate the local variations in the dryness of paint. The relationship between the optical signal obtained by the holographic method and the actual microscopic variations occurring in the paint film is also investigated using the gravimetric technique and a microscope. It is shown that the holographic technique can determine the stationary state of a painted surface corresponding to the end of the falling rate period in the drying process. The holographic technique detects mainly the activity on the surface and is applicable to assessment of the early drying process of paint.     

Method for determination of aflatoxin M₁ in cheese and butter by HPLC using an immunoaffinity column

A rapid, sensitive convenient method for determination of aflatoxin M? (AFM?) in cheese and butter by HPLC was developed and validated. The method employs a safe extraction solution (mixture of acetonitrile, methanol and water) and an immunoaffinity column (IAC) for clean-up. Compared with the widely used method employing chloroform and a Florisil column, the IAC method has a short analytical time and there are no interference peaks. The limits of quantification (LOQ) of the IAC method were 0.12 and 0.14 μg/kg, while those of the Florisil column method were 0.47 and 0.23 μg/kg in cheese and buffer, respectively. The recovery and relative standard deviation (RSD) for cheese (spiked at 0.5 μg/kg) in the IAC method were 92% and 7%, respectively, while for the Florisil column method the corresponding values were 76% and 10%. The recovery and RSD for butter (spiked at 0.5 μg/kg) in the IAC method were 97% and 9%, and those in the Florisil method were 74% and 9%, respectively. In the IAC method, the values of in-house precision (n=2, day=5) of cheese and butter (spiked at 0.5 μg/kg) were 9% and 13%, respectively. The IAC method is superior to the Florisil column method in terms of safety, ease of handling, sensitivity and reliability. A survey of AFM? contamination in imported cheese and butter in Japan was conducted by the IAC method. AFM? was not detected in 60 samples of cheese and 30 samples of butter.     

Profile of non-volatiles in whisky with regard to superoxide dismutase activity

SOD (Superoxide dismutase)-like activities of 23 kinds of single malt whisky (Scotch and Japanese) were evaluated. There was a positive correlation between SOD-like activity and the maturation age of whisky that exceeded the difference resulting from the manufacturing region. The SOD-like activity of Yamazaki 18, a typical single malt whisky in Japan, was approximately 1333 U/ml and that of non-volatile components in the whisky was 388U/mg, indicating that single malt whisky generally has a very strong SOD-like activity. To elucidate their contribution to SOD-like activity, the non-volatile components of whisky (Yamazaki 18) were ultrafiltered and separated with a Diaion HP20/water-EtOH system. Elution of the fraction less than 5000 molecular weight (<5000 MW fraction) with 60% (v/v) EtOH contributed most to SOD-like activity of the whisky. As this elution contained a considerable amount of polyphenolics, the content and SOD-like specific activity of ellagic acid, gallic acid, and lyoniresinol--the main whisky polyphenolics--were evaluated. The contribution of these compounds to the SOD-like activity of whisky was approximately 15%. Polyphenolics in whisky were relatively distributed to a higher MW fraction compared to carbohydrates in whisky, and specific activity (SOD-like activity per weight) of the >10,000 MW fraction was greater than that of the <5000 MW fraction, although the content of this fraction was low. These results indicate that various polyphenolics with higher molecular weights also contribute to the SOD-like activity of whisky together with main whisky polyphenolics.     

Synthesis of an agarose-gelatin conjugate for use as a tissue engineering scaffold

We synthesized a conjugate in which gelatin was covalently crosslinked to agarose using 1,1-carbonyldiimidazole (CDI) in dimethyl sulfoxide in order to obtain gels with cellular adhesiveness that showed a sol-to-gel transition, but no gel-to-sol transition, around body temperature. The gelatin content of the conjugate increased by 2.7-fold when the concentration of CDI was increased from 1.3 to 32.7 mM. Aqueous solutions of the conjugate gelled upon cooling from 40 degrees C to 20 degrees C, but did not remelt at 37 degrees C. The percentage of adhered cells after 4 h of culture on a gel created from a conjugate containing about 25 wt% gelatin was similar to that for cells grown on tissue culture dishes. The adhered cells proliferated on the conjugate gel during culture for a further 5 d. In addition, the conjugate used in this study did not result in mechanical instability of the resultant gel compared to that of an unmodified agarose gel. These results demonstrate that this agarose-gelatin conjugate is a good candidate material for tissue engineering.     

Elucidation of the effect of welding speed on melt flows in high-brightness and high-power laser welding of stainless steel on basis of three-dimensional X-ray transmission in situ observation

This research was performed with the objective of clarifying the effect of welding speed on melt flows during melt-run welding of SUS304 stainless steel plates with a 6-kW power laser beam on the basis of three-dimensional X-ray transmission in situ observation. As welding speed increased from 25 to 250 mm/s, three kinds of welds characterized by porosity formation and no defects or underfilling due to spatters were produced. The average and the maximum values of measured melt flow velocity were 3 and 10 times higher than the welding speed, respectively. Two kinds of circulation flows at the inlet or the tip of a keyhole were confirmed to control heat transfer in a molten pool. It was found that the circulation flows were so sensitive to the welding speed that bubbles resulting in porosity or spatters were often formed. According to X-ray observation of the formation of spatters with tungsten carbide (WC) tracers, as the melt flow rose along the keyhole wall, the velocity was accelerated from 0.24–0.54 m/s near the keyhole inlet. Consequently, the melt flows made the convex surface behind the keyhole grow higher, resulting in spattering.     

Novel Inhibitor for Downstream Targeting of Transforming Growth Factor-β Signaling to Suppress Epithelial to Mesenchymal Transition and Cell Migration

Cancer metastasis accounts for most of the mortality associated with solid tumors. However, antimetastatic drugs are not available on the market. One of the important biological events leading to metastasis is the epithelial to mesenchymal transition (EMT) induced by cytokines, namely transforming growth-factor-β (TGF-β). Although several classes of inhibitors targeting TGF-β and its receptor have been developed, they have shown profound clinical side effects. We focused on our synthetic compound, HPH-15, which has shown anti-fibrotic activity via the blockade of the TGF-β Smad-dependent signaling. In this study, 10 μM of HPH-15 was found to exhibit anti-cell migration and anti-EMT activities in non-small-cell lung cancer (NSCLC) cells. Although higher concentrations are required, the anti-EMT activity of HPH-15 has also been observed in 3D-cultured NSCLC cells. A mechanistic study showed that HPH-15 inhibits downstream TGF-β signaling. This downstream inhibition blocks the expression of cytokines such as TGF-β, leading to the next cycle of Smad-dependent and -independent signaling. HPH-15 has AMPK-activation activity, but a relationship between AMPK activation and anti-EMT/cell migration was not observed. Taken together, HPH-15 may lead to the development of antimetastatic drugs with a new mechanism of action.     

The prognosis of patients with gastric cancer possessing sex hormone receptors

Estrogen receptors (ER) and progesterone receptors (PgR) were immunohistologically investigated in 107 patients with gastric cancer who underwent curative resection. Both ER and PgR were detected only in the cancer cell nucleus. The ER positive rate was 27.7% for males and 31.0% for females, while the PgR positive rate was 9.2% for males and 11.9% for females. Clinicopathologically, the ER positive rate was slightly higher in young females and in cases of poorly differentiated gastric cancer. When cumulative survival rates were analyzed in relation to the presence or absence of receptors, the 10-year cumulative survival rate after surgery was significantly lower in the ER positive cases, being 15.7% cent, than in the ER negative cases, being 62.7%, and also significantly lower in the PgR positive cases, being 18.2%, than in the PgR negative cases, being 48.3%. The coexistence of ER and PgR in gastric cancer tissues suggests that the ER is physiologically active, or that ER positive gastric cancer is hormone-dependent. The poor prognosis of patients with receptor positive gastric cancer suggests that gastric cancer with these receptors is highly malignant.     

MORI-A CPS: 3D printed soft actuators with 4D assembly simulation

Artificial Life and Robotics - Soft modular robotics combines soft materials and modular mechanisms. We are developing a vacuum-driven actuator module, MORI-A, which combines a 3D-printed flexible...     

Hardenability Improvement Mechanisms and Hardenability-Controlling Factors in Steels With Combined Mo–B Addition

Metallurgical and Materials Transactions A - To elucidate the hardenability improvement mechanisms and hardenability-controlling factors in low-carbon (C) steels with the combined addition of...