Possible Biosynthetic Pathways for all <Emphasis Type="Italic">cis</Emphasis>-3,6,9,12,15,19,22,25,28-Hentriacontanonaene in Bacteria

A very long chain polyunsaturated hydrocarbon, hentriacontanonaene (C31:9), was detected in an eicosapentaenoic acid (EPA)-producing marine bacterium, which was isolated from the mid-latitude seashore of Hokkaido, Japan, and was tentatively identified as mesophilic Shewanella sp. strain osh08 from 16S rRNA gene sequencing. The geometry and position of the double bonds in this compound were determined physicochemically to be all cis at positions 3, 6, 9, 12, 15, 19, 22, 25, and 28. Although C31:9 was detected in all of the seven EPA- or/and docosahexaenoic acid-producing bacteria tested, an EPA-deficient mutant (strain IK-1Δ8) of one of these bacteria had no C31:9. Strain IK-1Δ8 had defects in the pfaD gene, one of the five pfa genes responsible for the biosynthesis of EPA. Although Escherichia coli DH5α does not produce EPA or DHA inherently, cells transformed with the pfa genes responsible for the biosynthesis of EPA and DHA produced EPA and DHA, respectively, but not C31:9. These results suggest that the Pfa protein complex is involved in the biosynthesis of C31:9 and that pfa genes must not be the only genes responsible for the formation of C31:9. In this report, we determined for the first time the molecular structure of the C31:9 and discuss the possible biosynthetic pathways of this compound.     

Multiple scattering effects of MeV electrons in very thick amorphous specimens

Multiple scattering has an important influence on the analysis of microns-thick specimens with MeV electrons. In this paper, we report on effects of multiple scattering of MeV electrons on electron transmission and imaging of tilted and thick amorphous film specimens by experiment and theoretical analysis. Electron transmission for microns-thick epoxy-resin and SiO₂ specimens calculated by the multiple elastic-scattering theory is in good agreement with measurements in the ultrahigh voltage electron microscope (ultra-HVEM) at Osaka University. Electron transmission and electron energy are then presented in an approximate power law. The bright-field ultra-HVEM images of gold particles on the top or bottom surfaces of 5 and 15 μm thick specimens further illustrate the effect of multiple scattering on image quality. The observed top‐bottom effect for the very thick specimens appears to be mainly caused by multiple elastic scattering. With increase in the accelerating voltage from 1 to 2 MV, image blurring, contrast, the signal-to-noise ratio, and the top‐bottom effect are improved because of reduction in the influence of multiple scattering. However, the effect of specimen thickness on image blurring is shown to be stronger than that of accelerating voltage. At the 2 MV accelerating voltage, the 100 nm gold particle can be imaged with less blurring of ∼4 nm when located at the bottom surface of a 15 μm thick epoxy-resin specimen.     

Practical cell labeling with magnetite cationic liposomes for cell manipulation

Personalization of the cell culture process for cell therapy is an ideal strategy to obtain maximum treatment effects. In a previous report, we proposed a strategy using a magnetic manipulation device that combined a palm-top size device and a cell-labeling method using magnetite cationic liposomes (MCLs) to enable feasible personalized cell processing. In the present study, we focused on optimizing the MCL-labeling technique with respect to cell manipulation in small devices. From detailed analysis with different cell types, 4 pg/cell of MCL-label was found to be obtained immediately after mixing with MCLs, which was sufficient for magnetic cell manipulation. The amount of label increased within 24 h depending on cell type, although in all cases it decreased along with cell doubling, indicating that the labeling potential of MCLs was limited. The role of free MCLs not involved in labeling was also investigated; MCLs' role was found to be a supportive one that maximized the manipulation performance up to 100%. We also determined optimum conditions to manipulate adherent cells by MCL labeling using the MCL dispersed in trypsin solution. Considering labeling feasibility and practical performance with 10³–10⁵ cells for personalized cell processing, we determined that 10 μg/ml of label without incubation time (0 h incubation) was the universal MCL-labeling condition. We propose the optimum specifications for a device to be combined with this method.     

Source/drain junction fabrication for Ge metal-oxide-semiconductor field-effect transistors

We established fabrication methods for high-quality Ge n⁺/p and p⁺/n junctions using thermal diffusion of P and implantation of B, respectively. The carrier concentrations in n⁺ and p⁺ layers were as high as 4 × 10¹⁹ and 2 × 10¹⁹ cm^− 3, respectively. It was found that a peripheral surface-state current dominates the reverse leakage current in an n⁺/p junction diode. The protection of junction surfaces from plasma damage during the SiO₂ deposition was essential to achieve high-quality source/drain junctions. The surface passivation with a GeO₂ interlayer was harmful to an n-channel metal-oxide-semiconductor field-effect transistor (MOSFET) because of an increase in a surface leakage current due to inversion carriers. For a p-channel MOSFET, on the other hand, the GeO₂ interlayer plays a role in decreasing the surface leakage current.     

One-pass separation of single-wall carbon nanotubes by gel chromatography with a gradient of surfactant concentration

We have investigated the diameter-selective separation of carbon nanotubes by one-pass gel chromatography with a gradient of surfactant concentration. The formation of surfactant gradient in a column was successfully measured and is explained by a simple diffusion process even in the gel. We found that the diameter of eluted nanotubes is inversely proportional to the surfactant concentration of eluate. The detailed analysis of the movement of the nanotubes in the gel revealed that the separation mechanism was qualitatively explained by a model based on the trapping and de-trapping events of the nanotube–surfactant micelle on the gel surface,where the probability of the trapping and de-trapping events is proportional to the product of the diameter of the nanotubes and the surfactant concentration.     

Direct plasma metabolite analysis of positron emission tomography radioligands by micellar liquid chromatography with radiometric detection

Determination of radio-metabolites in plasma samples taken during a positron emission tomography (PET) study is an important component in the pharmacokinetic evaluation of PET radioligands. We have developed and validated a new analytical procedure for the plasma metabolite analysis of PET radioligands based on micellar liquid chromatography using an anionic surfactant mobile phase. Chromatographic separation was performed on an octadecyl semipreparative column (10 mm I.D. × 160 mm, 10 μm) using 100 mM sodium dodecyl sulfate (SDS) and 1-butanol in 10 mM sodium-phosphate (pH 7.2) at a flow rate of 5 mL/min. The samples taken from monkey or human plasma during PET measurements were directly injected into a liquid chromatographic (LC) system coupled to an online radiometric detector under micellar conditions using 1-2% (v/v) 1-butanol mobile phase to remove plasma proteins and concentrate the analytes at the column head. At 2 min, mobile phase was changed to elute and separate PET radioligand and its radiometabolites with high peak capacity under high submicellar conditions (10-25% 1-butanol). This procedure allowed direct plasma injection (up to 2 mL) into the LC column without any pretreatment with a short analysis-time of 8-10 min. Satisfactory reproducibility, linearity, sensitivity, accuracy and recovery were obtained in the validation study. The developed method was successfully applied to study the metabolism for diverse groups of PET radioligands and provided reliable determination of PET radioligands in human and monkey plasma. This method is advantageous in terms of simplifying and shortening the processes required to analyze short-lived radioligands as well as in providing a more accurate estimation of the metabolite corrected input function, especially for the radioligands with lower recoveries or degradation potential during the deproteination process in a conventional procedure.     

Combination Treatment of OSI-906 with Aurora B Inhibitor Reduces Cell Viability via Cyclin B1 Degradation-Induced Mitotic Slippage

Insulin-like growth factor 1 receptor (IGF1R), a receptor-type tyrosine kinase, transduces signals related to cell proliferation, survival, and differentiation. We recently reported that OSI-906, an IGF1R inhibitor, in combination with the Aurora B inhibitor ZM447439 suppresses cell proliferation. However, the mechanism underlying this suppressive effect is yet to be elucidated. In this study, we examined the effects of combination treatment with OSI-906 and ZM447439 on cell division, so as to understand how cell proliferation was suppressed. Morphological analysis showed that the combination treatment generated enlarged cells with aberrant nuclei, whereas neither OSI-906 nor ZM447439 treatment alone caused this morphological change. Flow cytometry analysis indicated that over-replicated cells were generated by the combination treatment, but not by the lone treatment with either inhibitors. Time-lapse imaging showed mitotic slippage following a severe delay in chromosome alignment and cytokinesis failure with furrow regression. Furthermore, in S-trityl-l-cysteine–treated cells, cyclin B1 was precociously degraded. These results suggest that the combination treatment caused severe defect in the chromosome alignment and spindle assembly checkpoint, which resulted in the generation of over-replicated cells. The generation of over-replicated cells with massive aneuploidy may be the cause of reduction of cell viability and cell death. This study provides new possibilities of cancer chemotherapy.     

A New Space-Time Multiple Trellis Coded Modulation Scheme Using Transmit Symbol Phase Rotation

Space-time multiple trellis coded modulation (ST-MTCM) has been introduced in order to achieve maximum transmit diversity gain and larger coding gain with the existence of parallel paths. In our previous research, we designed a new coded modulation scheme for ST-MTCM which simultaneously maximizes the coding gain and diversity gain utilizing Hadamard Matrix giving the maximum determinant. This scheme, however, cannot achieve full transmit rate. In this paper, we extend our research so as to achieve full rate transmission as well as maximum coding and diversity gain. In addition, Super-Orthogonal Space-Time Trellis Code (SO-STTC) is well known for its high coding gain, full diversity gain and full transmit rate. Even though our proposed scheme is essentially the same as SO-STTC, we show in this paper that our proposed code design is different from SO-STTC, and achieves better performance. Our proposed code design utilizes transmit symbol phase rotation at a certain time slot so as to avoid same path transition in trellis, which occurs with conventional SO-STTC scheme. We design codes with different way of phase rotation for different MPSK modulation scheme, and simulation results show the improvements of our proposed codes for MPSK modulation with different number of states. Susu Jiang was born in Jilin, China, in 1979. She received the B.E. and M.S. degrees in electrical and computer engineering from Yokohama National University, Yokohama, Japan, in 2001 and 2003, respectively. She is currently working toward the Ph.D. degree in electrical and computer engineering at Yokohama National University, Yokohama, Japan. Her research interests include space-time coding, channel coding in wireless communications, and information theory. She is a student member of the IEICE and IEEE. Ryuji Kohno received the Ph.D. degree from the University of Tokyo in 1984. Dr. Kohno is currently a Professor of the Division of Physics, Electrical and Computer Engineering, Yokohama National University. In his currier he was a director of Advanced Telecommunications Laboratory of SONY CSL during 1998–2002 and currently a director of UWB Technology institute of National Institute of Information and Communications Technology (NICT). In his academic activities, he was elected as a member of the Board of Governors of IEEE Information Theory (IT) Society in 2000 and 2003. He has played a role of an editor of the IEEE Transactions on IT, Communications, and Intelligent Transport Systems (ITS). He is a fellow of IEICE, vice-president of Engineering Sciences Society of IEICE and has been the Chairman of the IEICE Technical Committee on Spread Spectrum Technology, that on ITS, and that on Software Defined Radio (SDR). Prof. Kohno has contributed for organizing many international conferences, such as an chair-in honor of 2002 & 2003 International Conference of SDR (SDR'02 & SDR'03), a TPC co-chair of 2003 International Workshop on UWB Systems (IWUWBS'03), and a general co-chair of 2003 IEEE International Symposium on IT (ISIT'03), that of Joint UWBST&IWUWB'04 and so on. He was awarded IEICE Greatest Contribution Award and NTT DoCoMo Mobile Science Award in 1999 and 2002, respectively.     

Model analysis of an inter-industrial and inter-regional waste recycling system in Japan

In this study, we investigate an inter-industrial and inter-regional recycling system for industrial waste by the cement industry in Japan. We develop a linear programming model that represents cement production processes and waste transportation of all cement factories in Japan. We simulate cost and CO₂-minimizing systems. The result implies that making waste transportation more efficient in cost is an effective means for CO₂ reduction.     

Hidden Markov model-based prediction of antigenic peptides that interact with MHC class II molecules

Elucidating the interaction between major histocompatibility complex (MHC) molecules and antigenic peptides is fundamental to better understanding of the processes involved in immune responses and for the development of innovative immunotherapies. In the present study, hidden Markov models (HMM) were combined with the successive state splitting (SSS) algorithm for optimization of the HMM structure, to predict peptide binders to the human MHC class II molecule HLA-DRB1*0101. The predictive performance of our model (S-HMM) was compared with fully connected HMM and artificial neural network (ANN) methods using the relative operating characteristic (ROC) analysis. The S-HMM predictions had values of ROC > or = 0.85 which was at least as good, or better than the comparison methods. In addition, S-HMM is trained on positive data only and does not require exhaustive data preprocessing, such as peptide alignment. Our results demonstrated that S-HMM combines the high accuracy of predictions with the simplicity of implementation and is therefore useful for analyzing MHC class II binding peptides. In particular the S-HMM may be trained using only positive data and, the preprocessing of training data, such as peptide alignment and the selection of binding cores, is not required in this method.