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Chinese hamster ovary (CHO) cells are widely used for constructing expression systems to produce therapeutic proteins. However, the establishment of high-producer clones remains a laborious and time-consuming process, despite various progresses having been made in cell line development. We previously developed a new strategy for screening high monoclonal antibody (mAb)-producing cells using flow cytometry (FCM). We also reported that p180 and SF3b4 play key roles in active translation on the endoplasmic reticulum, and that the productivity of secreted alkaline phosphatase was enhanced by the overexpression of p180 and SF3b4. Here, we attempted to apply the translational enhancing technology to high mAb-producing cells obtained after high-producer cell sorting. A high mAb-producing CHO clone, L003, which showed an mAb production level of3 g/L in fed-batch culture, was established from a high mAb-producing cell pool fractionated by FCM. Clones generated by the overexpression of p180 and SF3b4 in L003 cells were evaluated by fed-batch culture. The specific productivity of clones overexpressing these two factors was ∼3.1-fold higher than that of parental L003 cells in the early phase of the culture period. Furthermore, the final mAb concentration was increased to 9.5 g/L during 17 days of fed-batch culture after optimizing the medium and culture process. These results indicate that the overexpression of p180 and SF3b4 would be promising for establishing high-producer cell lines applicable to industrial production.

Polysorbate 80 (PS80) is an amphipathic, nonionic surfactant commonly used in pharmaceutical protein formulations and is composed of fatty acid (FA) esters of polyethoxylated sorbitan. However, commercial PS80 products contain substantial amounts of byproducts. The development of simple and reliable methods for PS80 component analysis is challenging given the inherent heterogeneity.We developed a comprehensive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to profile the components of PS80. Semi-comprehensive LC-MS/MS analyses of 11 subspecies in three commercial PS80 products were performed to estimate the average degree of polymerization of the ethylene oxide units (Avg-n) in the molecules. Furthermore, three subspecies (polyoxyethylene sorbitan monoester, polyoxyethylene isosorbide monoester, and polyoxyethylene monoester) were analyzed to estimate the composition ratios of the seven ester-bonded FAs present in PS80.The Avg-n values of five polyoxyethylene sorbitan esters (none, mono, di, tri, and tetra), three polyoxyethylene isosorbide esters (none, mono, and di), and three polyoxyethylene esters (none, mono, and di) were 26.5-30.6, 12.1-14.6, and 11.4-15.8, respectively. These values were comparable regardless of the number of ester-bonded FAs. Each product had a similar FA composition ratio regardless of the differences in the subspecies. However, the obtained C18:2 values were higher than those reported in the product certificates.The proposed LC-MS/MS method evaluated the overall PS80 components, revealing the possibility of underestimation of ester-bonded linoleic acid by the conventional GCMS method. The similarity of Avg-n values and FA compositions amang subspecies suggested the high reliability of these results, indicating that the presented approach may help in the quality control of PS80 formulations.

Aiming realization of a carbon circulation society, we considered a social model in which the carbon dioxide (CO2) can be used effectively by recovering CO2 from the air and converting it to valuables. To realize Direct Air Capture (DAC), we developed amines for capturing low-concentration CO2, and developed a system applying the CO2 capture technology developed until now by Kawasaki Heavy Industries, Ltd. (Kawasaki CO2 Capture, KCC). Additionally, we performed a survey on the technology for the effective use of CO2 applied to DAC as well as constructing a carbon circulation society model, and we also created a lifecycle assessment model (LCA) to evaluate the effectiveness of this social model using a carbon life cycle assessment model. This project will be conducted with the consignment of the Ministry of the Environment, Government of Japan for three years from 2019, and here we will report on the achievements of the first year.

A new synthetic polyamine was found as a suitable compound for amine-impregnated solid sorbent through screening of amine compounds with the aid of density functional theory calculations. CO 2 capture performance of the sorbent was evaluated on a lab-scale CO 2 capture system. The solid sorbent was excellently regenerable even at low temperature condition. Then, Bench-scale demonstration will be started on current project, using the sorbent developed by Research Institute of Innovative Technology for the Earth (RITE) and moving bed system of Kawasaki Heavy Industries, Ltd (KHI).

The Kawasaki CO2 Capture (KCC) system involves a solid adsorbent method that captures CO2 in exhaust gases using low-temperature steam; thus, it can use waste heat easily and save more energy than conventional technology. In this paper, we establish a design method for a large-scale plant using the KCC moving-bed method, and conduct parametric testing using a 5 ton/d bench scale test plant to find the characteristics of the moving-bed method. Using the relationship between the residence time of the adsorbent material and the CO2 adsorption load, we found that design specifications could be applied to a large-scale plant.

To screen a high-producing recombinant Chinese hamster ovary (CHO) cell from transfected cells is generally laborious and time-consuming. We developed an efficient enrichment strategy for high-producing cell screening using flow cytometry (FCM). A stable pool that had possibly shown a huge variety of monoclonal antibody (mAb) expression levels was prepared by transfection of an expression vector for mAb production to a CHO cell. To enrich high-producing cells derived from a stable pool stained with a fluorescent-labeled antibody that binds to mAb presented on the cell surface, we set the cell size and intracellular density gates based on forward scatter (FSC) and side scatter (SSC), and collected the brightest 5% of fluorescein isothiocyanate (FITC)-positive cells from each group by FCM. The final product concentration in a fed-batch culture of cells sorted without FSC and SSC gates was 1.2-1.3-times higher than that of unsorted cells, whereas that of cells gated by FSC and SSC was 3.4-4.7-fold higher than unsorted cells. Surprisingly, the fraction with the highest final product concentration indicated the smallest value of FSC and SSC, and the middle value of fluorescence intensity among all fractionated cells. Our results showed that our new screening strategy by FCM based on FSC and SSC gates could achieve an efficient enrichment of high-producing cells with the smallest value of FSC and SSC.

In a Kawasaki CO2 capture (KCC) system, CO2 in an exhaust gas is captured using a solid sorbent system with low-temperature steam. Because this system reuses the waste heat, it conserves energy in comparison with other conventional technologies. While a moving bed system makes it possible to apply the KCC technology to a large-scale plant, technology development at a large test facility is necessary. Therefore, this study investigates the application of the solid sorbent developed by Research Institute of Innovative Technology for the Earth (RITE) to the KCC moving-bed system for practical use. This study also evaluates the performance of the solid sorbent in a bench-scale test plant and preliminarily examines the installation at a 40 t-CO2/d-scaled pilot test plant to identify technological challenges at coal-fired power stations.

We compared reactivity between CO2 and Li2CuO2 with that between CO2 and Li4SiO4 by using XRD, SEM observation, and thermogravimetry in order to study fundamentally a potential of Li2CuO2 oxide for application to solid CO2 absorbents. The CO2 absorption ability of Li2CuO2 was examined by measuring rate constants for the gas–solid reaction between perfectly single-phase Li2CuO2 and CO2 at various temperatures in 100 vol% CO2 atmosphere using the rate theory with thermogravimetry using rapid heating. The measured rate constants were compared with those reported previously for Li4SiO4 at various temperatures. Almost no difference was found between these rate constants of Li2CuO2 and Li4SiO4 at temperatures less than 660 °C, but at temperatures higher than 670 °C, the rate constants of Li2CuO2 were greater than those of Li4SiO4. These results demonstrate that Li2CuO2 can outstrip the ability of Li4SiO4 to absorb CO2. Moreover, different from Li4SiO4, the CO2 absorption ability of Li2CuO2 decays only slightly, even if the atmospheric CO2 concentration decreases.

Recently, generic object recognition that achieves human-like vision has being looked to for use in robot vision, automatic categorization of images, and image retrieval. In object recognition, semi-supervised learning, which incorporates a large amount of unsupervised training data (unlabeled data) along with a small amount of supervised data (labeled data), is regarded as an effective tool to reduce the burden of manual annotation. However, some unlabeled data in semi-supervised models contain outliers that negatively affect the parameter estimation during the training stage. Such outliers often cause an over-fitting problem especially when a small amount of training data is used. Furthermore, another problem that occurs when using the conventional methods is that when labeling an image based on super-pixel representation, the lack of discrimination of the image features and the scale variance of the objects decreases the recognition accuracy because the feature extraction is based on the mono-scale segmentation. In this paper, we propose an object recognition method for solving both problems. For the former problem, our method prevents the over-fitting associated with the semi-supervised based approach by using sparse representation to suppress existing outliers in the data. For the latter problem, we employ Tree Conditional Random Field to construct the hierarchical structure of an image. Experiment results using two datasets confirm the effectiveness of our method.

Two kinds of immobilized ruthenium complex with tetradentate BABP (6,6′-bis(benzoylamino)-2,2′-bipyridine) ligand are prepared. One is compound 1 (FSM-Ru) directly bound to silica of the FSM-surface through Ru–O–Si bond, and others are compounds 2 (FSM-Aps-Ru) and 3 (FSM-Ims-Ru) with monodentate nitrogen ligands bound to silica surface. These ruthenium complexes encapsulated into mesopore of FSM showed interesting activity for oxygen transfer reaction of cyclohexene using tert-butylhydroperoxide (TBHP) as an oxidant. The FSM-Ru catalyst ( 1 ) having oxygen donor gave 1,2-epoxycyclohexane (epoxide) selectively through stepwise addition of TBHP, although 2 and 3 having nitrogen donor gave 2-cyclohexen-1-yl tert-butyl peroxide (peroxide) mainly. This remarkable selectivity control in the oxygen transfer reaction is interpreted in terms of the trans -influence of the ruthenium complex with the axially coordinating monodentate ligand on the silica support. Moreover, the heterogeneous catalyst 1 was recycled and used three times in epoxidation of cyclohexene, since leaching of the ruthenium species from 1 was not observed under argon atmosphere.

Particle size dependence of CO2 absorption rate of powdered Li4SiO4 with four kinds of particle sizes was studied by SEM observation, particle size distribution measurement, and thermogravimetry. Average particle sizes of the powdered single phase Li4SiO4 samples were estimated about 26 μm, 34 μm, 53 μm, 70 μm, respectively by the particle size distribution measurement and SEM observation. The measurement of apparent rate constant k for the CO2 absorption reaction showed that k values of all the powdered Li4SiO4 samples increase with the increasing measurement temperature T, following the Arrhenius equation in the region of 620-660°C, while the k values abruptly increase deviating the Arrhenius equation over 670°C. It was also noted that the k values seem to depend on the particle size of the powdered Li4SiO4 samples, that is, the k values increase with the decreasing particle size. Moreover, difference between the k values of all the powdered samples at same temperature was found small in the region of 620-660°C, while the difference was found large over 670°C. For Li4SiO4 with smaller particle size, Δk = k700°C - k660°C was found larger, vice versa, the Δk was found smaller for Li4SiO4 with larger particle size. The values seem to correspond with reaction mechanism for CO2 absorption of Li4SiO4 particle.

Three new complexes with FeIII2(μ-O)(μ-OH) core surrounded with one, two, and three amino groups, that is, (6-amino-2-pyridylmethyl)bis(pyridylmethyl)amine (MAPA), bis(6-amino-2-pyridylmethyl)(pyri...

Ruthenium(II) complexes, [Ru(babp)(dmso)L], consisting of tetradentate square-planar ligand (babp = 6,6′-bis(benzoylamino)-2,2′-bipyridinato) and two monodentate axial ones, DMSO and L (L = dmso or...

The CO2 absorption property of Li4SiO4 was investigated by thermogravimetry. Two kinds of sintered Li4SiO4 samples were prepared at 700°C and 1000°C. The CO2 absorption rate of the powdered Li4SiO4 samples was measured in 100 vol% CO2 atmosphere at nine selected temperatures in the 620~700°C region, and the measured data were discussed by the rate theory. The CO2 absorption reaction of the Li4SiO4 would be a pseudo first order reaction. For these samples, apparent reaction rate constants k were measured at different measurement temperatures, and activation energies were estimated by the Arrhenius plot. For both the samples, the estimated activation energies were about 115 kJ/mol in the 620-660°C region and about 56 kJ/mol in the 680-700°C region. The large difference between these activation energies seems to depend on the surface state of the Li4SiO4 particle reacting with CO2.

Synthesis and CO2 absorption property of Li4TiO4 oxide were studied by X-ray powder diffraction and thermogravimetry. Samples were prepared by the solid state method using Li2CO3 and Anatase or Rutile TiO2 as starting reagents. From the results of XRD, it was found that single phase Li4TiO4 can be obtained from Anatase TiO2 and Li2CO3 with 1 : 2.1 molar ratio in an alumina cell in atmosphere of Ar : H2=80 vol% : 20 vol% gas mixture (150 mL/min of flow rate) by heat-treatment up to 1000°C with 5°C/min of heating rate. By the results of thermogravimetry, it was found that single phase Li4TiO4 can absorb CO2 in the temperature range of 300°C to 856°C, and the maximum absorption is 42 mass% at 856°C. From the results of XRD, LiTiOx and Li2CO3 phases were found in the Li4TiO4 sample after complete CO2 absorption. Therefore, it is supposed that following reaction occurs in the reaction process between Li4TiO4 and CO2. 42 mass% of the maximum absorption could be predicted on the basis of the reaction. Li4TiO4(s)+1.5CO2(g)↔LiTiOx(s)+1.5Li2CO3(s)+(2.5-x)/2 O2(g) x=2~2.5

A square planar iron complex with H2babp was synthesized and its coordination behaviors with monodentate axial ligands were studied. Cl– or imidazole ligand coordinated at the axial positions of the complex, but pyridine could not be introduced. The heterocyclic nitrogen donors, such as pyridine and imidazole, abstracted the amide protons of the ligand H2babp [1]. The oxidation reactivity was controlled by the presence or absence of amide protons of planar ligand H2babp rather than the axial ligands. The oxidation of cyclohexene by their complexes in the presence of PhIO proceeded catalytically when the iron complexes with protonated H2babp was used, while only a slight amount of oxidation products were obtained when the complexes with deprotonated babp2– were used.

A new diferrous complex with asymmetric coordination sites, [Fe2(μ-OBz)(L)](ClO4)2 (1) (L = N,N-bis(6-methyl-2-pyridylmethyl)-N’,N’-bis(6-pivalamido-2-pyridylmethyl)-1,3-diaminopropan- 2-ol), was synthesized, which was immobilized in the internal surface of mesoporous silica (FSM-16) to construct a functional material (1A). The reaction of 1 with O2 in acetone at –50°C generated a peroxo complex 2 [λmax = 589 nm (ε = 2200 M-1 cm-1), ν16O-16O = 890 and 908 cm-1 (ν18O-18O = 852 cm-1)]. The μ-peroxo complex 2 was decomposed by bubbling of Ar gas without releasing O2, but the reaction of 1A with O2 at ambient temperature also formed a peroxo complex (2A) without decomposition. The reflectance spectrum of 2A showed a band assignable to the LMCT band from peroxo to a Fe(III) ion at 588 nm. Compound 2A is very stable, whose half-life time is 45 min, in contrast to a short life of 2 (310 sec). 2A released dioxygen by heating up to 75°C in vacuo, which was regenerated by bubbling of dioxygen at ambient temperature.

RGB is the standard color format for most digital video input and output devices, such as display monitors. However, each component consisting of a natural image has a lot of redundancy. So, a color conversion, which is RGB to YCrCb, has been being used to reduce its redundancy. However, each component of YCrCb signals has no correlation under the higher compression rate, so compression performance is decreased. So, we propose a new color conversion. By implementing the proposed method, which exploits the correlations between the color components, the PSNR of traditional way is higher than that of proposed way under low compression performance. However, it can not be found that the difference of reconstructed images quality is invisible. And the PSNR of traditional way becomes worse quickly under a high compression performance, however, that of proposed way becomes worse slowly, therefore proposed way can produce a better quality of reconstructed image in a low bit-rate. As a result, the examples demonstrated that a better performance of color image coding is obtained.

The slant-shaped squish chamber recently developed by our company improves charging efficiency and raises compression ratio of its gasoline-engines. We clarified its knock suppression mechanism using rapid compression-and-expansion machine, autoignition simulation, and 3-dimensional turbulent flame propagation simulation. The results indicate the following: 1. Timing of flame acceleration is a key of knock suppression, that is, optimum timing of flame acceleration exists in the latter half of combustion, because expansion stroke suppresses the temperature rise in the end gas. This universal mechanism improves knock-limited ignition timing and compression ratio of the above engines. 2. A cause of the flame acceleration in the slant-shaped squish chamber is strengthened turbulence occurring in front of the flame and in reverse squish flow which separates at the squish lip. 3. The reverse squish flow, which is well known to be induced by the suction due to the larger expansion ratio of squish region than the other region of combustion chamber, is enhanced by the thermal expansion of burning mixture.

A miniature torque sensor based on magnetostrictive effect, has been developed to be mounted inside an automobile engine (TOYOTA 4A-FE; 1.6-liter 4-cylinder). The sensor was installed in a last main bearing of the engine crankshaft so as to directly detect the torque generated by engine. This sensor was composed of two magnetic head-type elements, which were placed opposite to each other through the crankshaft in order to eliminate the influence of the off-center motion of the shaft on the sensor output. Furthermore, a commercial crankshaft made of cast nodular iron (JIS standard: FCD70) was adopted, taking account of high sensor sensitivity and mechanical stiffness.It was confirmed by tests on an engine-dynamometer arrangement that this sensor could clearly detect instantaneous torque waveform with four peaks during two crankshaft rotations, corresponding to four cylinder combustions, and peculiar torque fluctuation due to engine lean burn. In addition, it was found that true torque produced by engine could be detected in in-vehicle experiment. This sensor is useful for various automobile applications.

Recently, the absorption chiller/heater is attracting interest because it delivers airconditioning without using fluorocarbons. Practically, it requires periodical or non-periodical maintenance. It is difficult to detect failures except in summertime with large refrigerant load and, moreover, human expertise is needed for discrimination of their origin. This paper reports on a system that automatically predicts the possibility of failures and discriminates their origins, regardless of the driving situation.

An enzymatic hydrolysate of wheat gluten was further digested in vitro with porcine pepsin and pancreatin to obtain an indigestible peptide. Indigestible pyroglutamyl peptide was isolated from the digest by strong cation-exchange, size-exclusion, and reversed-phase chromatographies. The pyroglutamyl peptide was digested with pyroglutamate aminopeptidase, and the digest was reacted with phenyl isothiocyanate. The resultant phenylthiocarbamyl (PTC) peptides were purified by reversed-phase HPLC by using binary gradient elution with ammonium acetate buffer, pH 6.0, and acetonitrile. The PTC peptides were analyzed with an automatic peptide sequencer on the basis of the Edman degradation method with a modified program. Some pyroglutamyl peptides were also analyzed by fast-atom bombardment ionization mass spectrometry without the pyroglutamate amino peptidase digestion. Consequently, pyroGlu-Asn-Pro-Gln, pyroGlu-Gln-Gln-Pro-Gln, pyroGlu-Gln-Pro-Gln, pyroGlu-Gln-Pro-Gly-Gln-Gly-Gln, pyroGlu-Gln, pyroGlu-Gln-Pro, pyroGlu-Ile-Pro-Gln, pyroGlu-Ile-Pro, pyroGlu-Gln-Pro-Leu, pyroGlu-Gln-Phe-Pro-Gln, pyroGlu-Ser-Phe-Pro-Gln, pyroGlu-Phe-Pro-Gln, and pyroGlu-Gln-Pro-Pro-Phe-Ser were identified.

Synthesis and CO2 absorption properties of single-phase Li2CuO2 oxide were studied by X-ray powder diffraction and thermogravimetry. Samples of Li2CuO2 were prepared in solid state using Li2CO3 and...

A novel iron complex with a distorted coordination geometry, [FeII((−)-sp)(OBz)2] (1) ((−)-sp = (−)-sparteine, OBz = benzoate anion), was prepared and structurally characterized by X-ray analysis. ...

The Ru complex, [RuII(babp)(dmso)2] (1) (H2babp = 6,6′-bis(benzoylamino)-2,2′-bipyridine), was immobilized into mesoporous silicate FSM-16. The heterogeneous catalyst demonstrated a more selective ...

Using the evaluation form of the SICOT-IDES (Societe Internationale de Chirurgie Orthopedique et de Traumatologie-International Documentation and Evaluation System), clinical evaluation and radiological evaluation were performed on 50 joints of South East Asia Pacific (SEAP) Total Knee Arthroplasty in 31 patients. The clinical results of the SEAP total knee series could be satisfactorily described in detail, and are easily utilized for further investigation.

Detection of surgically manipulated extension is sometimes difficult during skull-base surgery. It is, however, essential to preserve important structures and to excise the lesion precisely. To solve this problem we developed a frameless, armless neurosurgical navigational system (“CANS navigator”). The CANS navigator utilizes electromagnetic coupling technology and detects the spatial position and orientation angle of a suction tube attached to a magnetic sensor. It displays the tip of the suction tube as well as its direction at a real-time basis on the preoperative images. Because the suction tube is continually directed toward the point of surgical manipulation, the navigator is able to demonstrate the extent of the surgical manipulation by accumulating the manipulation points on the display (“surgical track”). Before the skin incision is made, this system is useful in optimizing the patient’s head-position and the microscope angle and in determining the extent of the skin incision. After the craniotomy, this system assists the operator in clarifying surgical orientation, in preserving important structures encased in the tumors, and in confirming the dissectable range of neoplasms with unclear or irregular margins. Surgical track display is especially powerful in recurrent surgery or staged surgery in which the normal anatomy has been destroyed. The CANS navigator introduces precise localization of CT or MR to conventional skull-base surgery without limiting the operative field or interfering with the surgical procedures and assists the surgeon in reducing surgical invasion.

Two genes, petB and petD , encoding cytochrome b 6 and subunit IV of the cytochrome b 6 /ƒ complex, respectively, of the chloroplast genome of Marchantia polymorpha , a liverwort, have been mapped between the psbH and rpoA genes and each of the petB and petD genes has been predicted to contain a group II intron in its coding sequence [(1986) Nature 322, 572–574]. Primer extension analysis of the liverwort chloroplast RNA using two synthetic oligodeoxyribonucleotides, complementary to parts of the coding sequences of the petB and petD transcripts, as primers has provided evidence that the two genes are co-transcribed to a precursor mRNA which is then precisely spliced at the predicted sites.

In the process of capturing CO 2 from flue gas (combustion exhaust gas), the lowering of CO 2 capture energy is considered a significant issue. If some or all of the CO 2 capture energy can be compensated with the waste heat, a significant energy saving is possible. In our proposed CO 2 adsorption process, because the CO 2 is captured using low-temperature steam, an energy-s ving process that makes it is easy to utilize the waste heat can be created. In this paper, we conduct bench tests aimed at developing a mo ing-bed system suitable for large-scale plants in order to verify the performance of the adsorbent. The results demonstrated that an moving-bed system could be established to capture 1.6t/day of CO 2 from coal combustion exhaust gas.

A general and versatile method for the preparation of a variety of β-hydroxybenzothiazoles (aldol-type adducts) by the reaction of α-lithio-2-methylbenzothiazole with carbonyl compounds was proposed. As an example of the use of α-lithio-2-methylbenzothiazole as a masked enolate, the product obtained by the reaction of this lithium reagent with 5-chloro-2-pentanone could be effectively transformed into the corresponding β-oxyaldehyde [2-methyl-2-(2-oxoethyl)-tetrahydrofuran].

A helical port with swirl control valve (SCV) has been developed to satisfy two inconsistent requirements of achieving sufficient swirl generation to improve the combustion and still maintaining high volumetric efficiency. Their effects on combustion were confirmed in a single cylinder engine using high speed flame photography and cylinder pressure diagram analysis which has demonstrated faster combustion. As a result of a hot wire anemometer study, the differences in gas motion were clarified between two helical port with and without SCV, and more active movement of the center of swirl was measured in the case of helical port with SCV which suggests generation of higher turbulence in the cylinder.