Molecular Mechanisms and Tumor Biological Aspects of 5-Fluorouracil Resistance in HCT116 Human Colorectal Cancer Cells

5-Fluorouracil (5-FU) is a cornerstone drug used in the treatment of colorectal cancer (CRC). However, the development of resistance to 5-FU and its analogs remain an unsolved problem in CRC treatment. In this study, we investigated the molecular mechanisms and tumor biological aspects of 5-FU resistance in CRC HCT116 cells. We established an acquired 5-FU-resistant cell line, HCT116R^F10. HCT116R^F10 cells were cross-resistant to the 5-FU analog, fluorodeoxyuridine. In contrast, HCT116R^F10 cells were collaterally sensitive to SN-38 and CDDP compared with the parental HCT16 cells. Whole-exome sequencing revealed that a cluster of genes associated with the 5-FU metabolic pathway were not significantly mutated in HCT116 or HCT116R^F10 cells. Interestingly, HCT116R^F10 cells were regulated by the function of thymidylate synthase (TS), a 5-FU active metabolite 5-fluorodeoxyuridine monophosphate (FdUMP) inhibiting enzyme. Half of the TS was in an active form, whereas the other half was in an inactive form. This finding indicates that 5-FU-resistant cells exhibited increased TS expression, and the TS enzyme is used to trap FdUMP, resulting in resistance to 5-FU and its analogs.     

Biosynthesis of Trehangelin in Polymorphospora rubra K07–0510: Identification of Metabolic Pathway to Angelyl‐CoA

Trehangelins are trehalose angelates discovered from endophytic actinomycete Polymorphospora rubra K07‐0510. We identified the trehangelin biosynthetic gene cluster, including genes that encode a glycoside hydrolase‐like protein (thgC), α‐amylase (thgD), 3‐ketoacyl‐ACP synthase III (thgI), 3‐ketoacyl‐ACP reductase (thgK), enoyl‐CoA hydratase (thgH) and acyl transferase (thgJ). Heterologous expression of thgH, thgI, thgJ and thgK confirmed the importance of these genes in the biosynthesis of trehangelin A. Enzymatic activity studies showed that ThgI catalyses the condensation of acetyl‐CoA and methylmalonyl‐CoA to 2‐methylacetoacetyl‐CoA (MAA‐CoA), ThgK catalyses NADPH‐dependent reduction of MAA‐CoA to 3‐hydroxy‐2‐methylbutyryl‐CoA (HMB‐CoA) and ThgH catalyses the dehydration of HMB‐CoA to angelyl‐CoA (AN‐CoA). This is the first report on the elucidation of the enzymatic formation of AN‐CoA.     

Contact force detection on a minute aperture mounted optical contact slider using an acoustic emission sensor

Advances in the digital network society require both higher density and higher transfer rates in all sorts of storage systems. Especially in optical recording, the trend toward higher density and larger capacity requires novel surface-recording technologies that can drastically diminish head-to-medium spacing, resulting in an improvement in spatial resolution and, finally, a higher recording density. To this end, we have already proposed a novel contact optical head slider that is able to almost cancel the suspension load by generating hydrodynamic pressure, thus realizing a lower net contact force. Evaluating the dynamic contact force is requisite in realizing its stable sliding operation and higher signal readout performance. In this study, a continuous acoustic emission (AE) signal was utilized to detect the dynamic contact force. AE signal modulation was compared with the applied out-of-plane acceleration on a medium generated by a spindle combined piezo-electric actuator. It was clarified that the detected AE modulation amplitude was approximately proportional to the applied acceleration, and that utilizing AE modulation will be a useful method for evaluating dynamic contact force.     

Hypothesis testing in rank-size rule regression

This note examines testing methods for Paretoness in the framework of rank-size rule regression. Rank-size rule regression describes a relationship found in the analysis of various topics such as city population, words in texts, scale of companies and so on. In terms of city population, it is basically an empirical rule that log?(_S(i))$\log ?(S_{(i)})$ is approximately a linear function of log?(i)$\log ?(i)$ where _S(i)$S_{(i)}$ is the number of population of i  th largest city in a country. This is closely related to the so-called Zipf’s law. It is known that this kind of empirical observation is found when the city population is a random variable following a Pareto distribution. Thus one may be willing to test if city size has a Pareto distribution or not. Rosen and Resnick [K.T. Rosen, M. Resnick, The size distribution of cities: an explanation of the Pareto law and primacy, Journal of Urban Economics 8 (1980), 165–186] and Soo [K.T. Soo, Zipf’s law for cities: a cross country investigation, Regional Science and Urban Economics (35) 2005, 239–263] regress log?(_S(i))$\log ?(S_{(i)})$ on log?(i)$\log ?(i)$ and log?²(i)${\log ?}^2(i)$ and test the null of Paretoness by standard t-test for the latter regressor. It is found that t-statistics take large values and the Paretoness is rejected in many countries. We study the statistical properties of the t-statistic and show that it explodes asymptotically, in fact, by simulation and thus the t-test does not provide a reasonable testing procedure. We propose an alternative test statistic which seems to be asymptotically normally distributed. We also propose a test with the null hypothesis that the city size distribution is Pareto with exponent unity, which is a modification of the F-test.     

Augmented reality-based block piling game with superimposed collapse prediction

Understanding what cannot be seen is difficult. Physical behavior can be explained on the basis of physical theories even if the behavior cannot be observed. Explanation of what is physically happening in the real world would become easy, however, if annotations were superimposed on the real objects. Herein, the authors demonstrate how an understanding of a physical event can be facilitated by overlapping a real-world situation with a simulation that predicts a future state. This idea is demonstrated in a game application in which a player stacks blocks into a pile until it collapses. In general, it is easy to estimate whether a block on the edge of a table will fall or not. However, it is more difficult to predict whether a stack of many blocks will collapse, and in what manner the stack will collapse. Even though previous research has demonstrated that the problem of how two-dimensionally stacked blocks collapse can be reduced to solving a sequence of convex quadratic programs, algorithms for convex quadratic programs require massive computational resources. Hence, the authors developed a fast and new algorithm based on a linear program. The proposed algorithm realizes real-time simulation based on physics that superimposes predicted collapse. The block that is predicted to fall is superimposed on the real block with a lit background projection. The system was evaluated in an experiment, and superimposed augmented reality annotation was observed to be efficient. The system was also demonstrated in game contests and received positive feedback and comments.     

Ultra micro glutamate sensor using platinized carbon-fiber electrode and integrated counter electrode

An integrated ultra micro glutamate sensor has been constructed with a 7 μm diameter platinized carbon-fiber disk (PCD) electrode and a platinum thin-film (PTF) counter electrode fabricated on the glass capillary tube. By platinization, the electrode activity of the carbon fiber is improved. In order to obtain a stable response, a pulse potential is applied for hydrogen peroxide measurement. The PTF counter electrode shows good stability and can be used as a substitute for a silver-silver chloride electrode. Since the integrated PCD electrode shows good characteristics as a hydrogen peroxide sensor, glutamate oxidase is immobilized onto the tip of the PCD electrode to construct the ultra micro glutamate sensor. The sensor shows stable response to glutamate and a response time within 12 s. The calibration range for glutamate measurement is 50–800 μM.     

Structural changes of glass-ceramics of the Cu2O-Al2O3-SiO2 system on heating in air

The behaviour of copper ions in low thermal expansion glass-ceramics prepared from Cu₂O · Al₂O₃ ·_nSiO₂ glasses, or in-spodumene type Cu₂O · Ae₂O₃ ·nSiO₂ crystals included in the glass-ceramics on heating in air was investigated. On hewing at 300 to 500° C, the copper ions behaved as in the corresponding glasses. Cuprous ions in the glass-ceramics or-spondumene type crystals were oxidized into the cupric state, and at the same time an equal amount of cuprous ions to those oxidized were expelled out of the specimen for the requirement of electrical charge neutrality and then reacted with oxygen to form CuO on the surface. The oxidation of cuprous ions and the decrease of the copper content of the-spondumene type crystals brought about a considerable decrease in the lattice spooings.     

Preparation of multiple oxide BaTiO3 fibres by the sol-gel method

Multiple oxide BaTiO₃ gel fibres were prepared by the sol-gel method from Ba(OC₂H₅)₂-Ti(O-isoC₃H₇)₄-H₂O-C₂H₅OH-CH₃COOH and Ba(CH₃COO)₂-Ti(O-isoC₃H₇)₄-H₂O-CH₃COOH solutions. Relatively long gel fibres of 10cm length were obtained from both solutions in the limited composition region. The latter solution in particular showed a spinnability even when it contained no water. Therefore, the occurrence of spinnability of the solution was considered to be due to the formation of linear polymers composed of bridging acetate groups such as TiO-C(CH₃)-O-Ti rather than metalloxane bonding as Ti-O-Ti. Addition of water to the solutions seems to break the bridging acetate bonds and replace some of them by bridging oxygen bonds. The as-drawn gel fibres which were X-ray amorphous crystallized into BaTiO₃ ceramic fibres of 5mm average length upon heating above 600 ° C. However, the gel fibres drawn from the sols without water became powdery on heating because of the lack of Ti-O-Ti metalloxane bonds. The crystallization behaviour of the BaTiO₃ gel fibres is discussed based on the infrared spectroscopy, X-ray diffraction analysis and thermogravimetric-differential thermal analysis.     

Lysophosphatidylinositol Induced Morphological Changes and Stress Fiber Formation through the GPR55-RhoA-ROCK Pathway

We previously reported that lysophosphatidylinositol (LPI) functions as an endogenous agonist of GPR55, a novel cannabinoid receptor. However, the physiological roles of LPI-GPR55 have not yet been elucidated in detail. In the present study, we found that LPI induced morphological changes in GPR55-expressing HEK293 cells. LPI induced the cell rounding of GPR55-expressing HEK293 cells but not of empty-vector-transfected cells. LPI also induced the activation of small GTP-binding protein RhoA and increased stress fiber formation in GPR55-expressing HEK293 cells. The inhibition of RhoA and Rho kinase ROCK by the C3 exoenzyme and the ROCK inhibitor reduced LPI-induced cell rounding and stress fiber formation. These results clearly indicated that the LPI-induced morphological changes and the assembly of the cytoskeletons were mediated through the GPR55-RhoA-ROCK pathway.