Reconstitution of hepatic tissue architectures from fetal liver cells obtained from a three-dimensional culture with a rotating wall vessel bioreactor

Reconstitution of tissue architecture in vitro is important because it enables researchers to investigate the interactions and mutual relationships between cells and cellular signals involved in the three-dimensional (3D) construction of tissues. To date, in vitro methods for producing tissues with highly ordered structure and high levels of function have met with limited success although a variety of 3D culture systems have been investigated. In this study, we reconstituted functional hepatic tissue including mature hepatocyte and blood vessel-like structures accompanied with bile duct-like structures from E15.5 fetal liver cells, which contained more hepatic stem/progenitor cells comparing with neonatal liver cells. The culture was performed in a simulated microgravity environment produced by a rotating wall vessel (RWV) bioreactor. The hepatocytes in the reconstituted 3D tissue were found to be capable of producing albumin and storing glycogen. Additionally, bile canaliculi between hepatocytes, characteristics of adult hepatocyte in vivo were also formed. Apart from this, bile duct structure secreting mucin was shown to form complicated tubular branches. Furthermore, gene expression analysis by semi-quantitative RT-PCR revealed the elevated levels of mature hepatocyte markers as well as genes with the hepatic function. With RWV culture system, we could produce functionally reconstituted liver tissue and this might be useful in pharmaceutical industry including drug screening and testing and other applications such as an alternative approach to experimental animals.     

Electrochemical DNA methylation detection for enzymatically digested CpG oligonucleotides

We describe the electrochemical detection of DNA methylation through the direct oxidation of both 5-methylcytosine (mC) and cytosine (C) in 5'-CG-3' sequence (CpG) oligonucleotides using a sputtered nanocarbon film electrode after digesting a longer CpG oligonucleotide with endonuclease P1. Direct electrochemistry of the longer CpG oligonucleotides was insufficient for obtaining the oxidation currents of these bases because the CG rich sequence inhibited the direct oxidation of each base in the longer CpG oligonucleotides, owing to the conformational structure and its very low diffusion coefficient. To detect C methylation with better quantitativity and sensitivity in the relatively long CpG oligonucleotides, we successfully used an endonuclease P1 to digest the target CpG oligonucleotide and yield an identical mononucleotide 2'-deoxyribonucleoside 5'-monophosphate (5'-dNMP). Compared with results obtained without P1 treatment, we achieved 4.4 times higher sensitivity and a wider concentration range for mC detection with a resolution capable of detecting a subtle methylated cytosine difference in the CpG oligonucleotides (60mer).     

Development of a new X-ray source using backscattered X-ray with the use of a cold cathode

The development of an intense X-ray source using backscattered X-ray produced using an advanced electrode configuration is described. The electrodes were composed of field emitters deposited on a wire mounted on a perforated plate as the cathode and a copper plate as the anode. Electrons from these emitters collided with the copper plate and X-ray was generated at collision points. The backscattered X-ray in the direction normal to the electron trajectory through a hole in the anode escaped from the vacuum chamber through a beryllium window. Continuous and characteristic X-rays were detected at an applied voltage lower than that of a conventional X-ray source from 3.0 to 9.4 kV, respectively. Moreover, the X-ray dosage measured with a survey meter reached 0.95 mSv/h at 5.0 kV of applied voltage. The transmission images of three types of material used as an X-ray source for the X-ray imaging system indicate three advantages; low power consumption, focal point controllable by adjusting applied voltage, and photographable motion picture of X-ray transmission.     

Electrical characterization of high-k gate dielectrics on Ge with HfGeN and GeO2 interlayers

Two kinds of HfSiO_x/interlayers (ILs)/Ge gate stack structures with HfGeN- and GeO₂-ILs were fabricated using electron cyclotron resonance (ECR) plasma sputtering and the subsequent post deposition annealing (PDA). It was found that HfGe was formed by the deposition of Hf metal on Ge and changed to HfGeN by N₂ ECR-plasma irradiation, which was used as IL. Another IL was GeO₂, which was grown by thermal oxidation at 500 °C. For dielectrics with HfGeN-IL, PDA of 550 °C resulted in effective oxide thickness (EOT) of 2.2 nm, hysteresis of 0.1 V, and interface state density (D_it) = 7 × 10¹² cm^− 2 eV^− 1. For dielectrics with GeO₂-IL, PDA of 500 °C resulted in EOT of 2.8 nm, hysteresis of 0.1 V, and D_it = 1 × 10¹² cm^− 2 eV^− 1. The structural change of HfSiO_x/GeO₂/Ge during the PDA was clarified by using X-ray photoelectron spectroscopy, and the gate stack formation for obtaining the good IL was discussed.     

Current control of a PWM power amplifier by an approximate 2-degree-of-freedom digital controller

There is a need for robust current control of a pulse width modulation (PWM) power amplifier whose transient response characteristics do not deteriorate with extensive load changes and/or direct-current power supply voltage changes. In this article, we propose a digital robust controller with bumpless mode switching to control the current of a PWM power amplifier to satisfy the demands and extend the range of an inductive load width. It is necessary to measure the value of the load in order to implement this bumpless mode switching automatically depending on the load range. Therefore, a method of estimating the inductive load is shown. The bumpless mode switching is automatically performed by estimating an inductive value without specifying the value of the inductive load beforehand. That is, the value of the inductive load is estimated during the DDC execution, and the control mode is automatically switched bumplessly according to this estimated value. A digital controller equipped with inductance estimation and bumpless mode switching is realized by a DSP. Some experiments show that the digital controller with the proposed bumpless mode switching can satisfy larger specifications.     

Development of a chirped pulse amplification laser with zigzag slab Nd:glass amplifiers dedicated to x-ray laser research

A chirped pulse amplification laser with zigzag slab Nd:glass amplifiers dedicated to x-ray laser research is described. The laser provides a 1.6 ps duration pulse with approximately 7 J energy at a repetition rate of 0.1 Hz. In the power amplifier system, laser light is amplified in a two-step manner: The first step is image-relayed multipass amplification up to approximately 1 J with a 10 mm x 10 mm beam. The second step is double-pass amplification up to >10 J with a 10 mm x 90 mm beam. By using this laser system, the saturated amplification of the Ni-like Ag laser at a wavelength of 13.9 nm has been successfully demonstrated.     

Vertical seismic response of overhead crane

Vertical seismic response behavior is an important issue for the seismic design of equipments. The equipment, which is comparatively soft and unrestrained vertically, may resonate and its response is significantly magnified under vertical seismic excitation. Overhead crane is an example of equipment that is unrestrained vertically. The dynamic behavior of an 150-ton-capacity overhead crane under vertical seismic excitation was investigated by scale model excitation test and nonlinear time history analysis. The excitation tests were performed with several input levels and the vertical response with each input level was obtained. The simulation analysis approximately corresponded to the results of the excitation test.     

Precise metabolic flux analysis of coryneform bacteria by gas chromatography-mass spectrometry and verification by nuclear magnetic resonance

Precise metabolic flux analysis (MFA) by gas chromatography-mass spectrometry (GC-MS) and computer calculation was performed, and the consistency of the estimated results was verified by independently performed nuclear magnetic resonance (NMR) analysis. The precise estimation of flux by the integration method of the mass isotopomer signal, defined as the coefficient of variance (CV) of multiple determination, was investigated, and the results estimated using different data sets with the same magnitude of error were confirmed. The CV of multiple determinations was sufficiently small to discuss and compare the fluxes of a metabolic pathway. The estimated fluxes using the GC-MS data were cross-validated with the NMR data that were independently measured and not used for MFA. The developed method was successfully applied to the MFA of the growth phase of two different glutamate-producing coryneform bacteria, Corynebacterium glutamicum and C. efficiens. The difference in the growth rate between these two bacterial species was discussed while considering the results of MFA, including forward and backward (exchange) fluxes.     

Occurrence of paralytic shellfish poison (PSP)-producing dinoflagellate Alexandrium tamarense in Hiroshima Bay, Hiroshima Prefecture, Japan, during 1993-2004 and its PSP profiles

To assess levels of shellfish intoxication by the paralytic shellfish poison (PSP)-producing dinoflagellate Alexandrium tamarense, potential health risks to human shellfish consumers and the possible need for regulatory intervention, yearly variations of maximum cell density of this species were examined from 1993 to 2004 in Kure Bay and Kaita Bay, which are located within Hiroshima Bay, Hiroshima Prefecture, Japan. The seawater temperature was determined concomitantly. In Kure Bay, maximum concentrations of 1,400 and 1,300 cells/mL at 0 and 5 m depths were observed on 21 and 24 April 1997. In Kaita Bay, remarkably high concentrations above 1,000 cells/mL of A. tamarense were observed in two out of three years investigated. These facts suggest that the environment in both bays is favorable for the propagation of A. tamarense. The temperature range at which the natural population of A. tamarense blooms was generally from 12 to 16 degrees C. Four strains (ATKR-94, -95, -97 and -01) from Kure Bay and one strain (ATKT-97) from Kaita Bay were established. The strain ATKR-94, cultured in modified SW-2 medium at 15 degrees C for 15 days, showed a specific toxicity of 33.8 x 10(-6) MU/cell. The toxins in all five strains exist almost exclusively as beta-epimers (C2 (PX2 or GTX8), GTX3, dcGTX3 and GTX4), which accounted for 54.9 to 73.0 mol% of the total. The corresponding a-epimers (C1 (PX1 or epi-GTX8), GTX2, dcGTX2 and GTX1) accounted for 6.0 to 28.9 mol%. The toxin profiles of ATKR-97 and ATKT-97 were characterized by unusually high proportions of low-potency sulfocarbamoyl toxin, which comprised 62.4 and 68.2 mol%, respectively, of total toxins. In the toxic bivalves, the low-toxicity sulfocarbamoyl components, major components of A. tamarense, were present in amounts of only a few percent, suggesting that in vivo conversion of PSP occurs after ingestion. A comparison of the toxin profiles of the causative dinoflagellate and contaminated bivalves showed that PSP components exist in the bivalves in the form of alpha-epimers, presumably owing to accumulation or storage of the toxins.