Visualization of mitochondria with green fluorescent protein in cultured fibroblasts from patients with mitochondrial diseases

cDNAs for green fluorescent protein (GFP) and for a GFP fusion protein containing the presequence of human ornithine transcarbamylase (pOTC-GFP) were transfected into cultured human fibroblasts. GFP cDNA gave diffuse fluorescence throughout the cytoplasm and the nucleus, whereas pOTC-GFP cDNA gave mitochondria-associated fluorescence. Fluorescent mitochondrial structures could be classified into five patterns: thread-like mitochondria, fine thread-like ones, rod-like ones, granular ones, and granular ones with weak cytosolic fluorescence. pOTC-GFP mutants resulted in a loss of mitochondrial fluorescence and an appearance of weak fluorescence throughout the cytoplasm. pOTC-GFP cDNA was transfected into fibroblasts from patients with various mitochondrial diseases. Higher ratios of fibroblasts with granular mitochondria and those with fine thread-like ones were observed in a patient with Reye's syndrome and a patient with Kearns-Sayre syndrome. Weak cytosolic fluorescence was sometimes observed in fibroblasts from these patients. This method will be useful to analyze mitochondrial structural alterations and disorders of mitochondrial protein import.     

Practical method for generation expansion planning based on dynamic programming

This paper presents an efficient method which provides the optimal generation mix and the optimal generation construction process. The approximation method in which the dynamic programming technique and gradient method are combined is applied to determine the optimal generation mix with hydropower generation technologies. The successive approximations dynamic programming (SADP) technique, which is very suitable for high-dimensional multistage decision process problems, is used for obtaining the optimal generation construction process. The effectiveness and feasibility of the developed technique are demonstrated on a practical power system model which has five types of generation technologies including a hydropower generation technology.     

Effects of nondensifying inclusions on the densification and microstructure of zinc oxide matrix composites

The densification and microstructure development of ZnO containing Zn₇Sb₂O₁₂, ZrO₂, and aggregated ZnO were investigated to elucidate the effect of nondensifying inclusions on the sintering of ceramic/ceramic composites. The inclusion retarded the densification, and the degree of retardation was found to depend on the chemical species of inclusion; Zn₇Sb₂O₁₂ had the largest effect, followed by ZrO₂ and then aggregated ZnO last. The experimental results for aggregated ZnO was explained by the theory which predicts the generation of backstresses. The backstresses give a less significant effect on the densification. For Zn₇Sb₂O₁₂ and ZrO₂, the microstructure of the matrix varied with distance from an inclusion particle; much porosity was observed in the region surrounding the inclusion. Circumferential voids, which are responsible for the suppression of densification, form during the initial stage of sintering. Inclusion particles generate an anchoring effect which retards the densification of the matrix immediately surrounding the inclusion particle during the intermediate stage.Supported by the Inamori Foundation.     

Gel chromism and anomalous luminescence in poly(3-alkylthiophene)

The absorption spectrum of poly(3-alkylthiophene) gel changes drastically in association with a volume instability upon changing the solvent composition and also the temperature. In the expanded state in chloroform and at high temperature, the photoluminescence is much enhanced compared with the shrunken state in ethanol and at low temperature. This gel chromism and anomalous luminescence are discussed in terms of the change of effective conjugation length induced by the steric hindrance and the dynamics of the photoexcited species.     

Interconnected power systems with superconducting magnetic energy storage

The objective of this work is to discuss the concept of back‐to‐back interconnection systems with energy storage, especially with a Superconducting Magnetic Energy Storage (SMES) incorporated into a back‐to‐back DC link. In this case, each converter of the back‐to‐back system is used as a power conditioning system for the SMES coils. Since the AC–DC converter can be designed independently of the frequency of the power system, a two‐way switch is connected to the AC side of each converter. This two‐way switch can select the interconnected power systems. By using the two‐way switches, this system can provide the stored energy in the SMES system to each interconnected power system through two AC–DC converters. For instance, lower‐cost power of each power network can be stored through two converters during the off‐peak hours and made available for dispatch to each power network during periods of demand peak. Then this system increases the reliability of electric power networks and enables the economical operations depending on the power demand. This paper describes the unique operations of the back‐to‐back interconnection with SMES and discuses the optimal SMES configuration for a 300‐MW‐class back‐to‐back interconnection. © 2008 Wiley Periodicals, Inc. Electr Eng Jpn, 164(2): 37–43, 2008; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.20482     

A Novel Artificially Humanized Anti-Cripto-1 Antibody Suppressing Cancer Cell Growth

Cripto-1 is a member of the EGF-CFC/FRL1/Cryptic family and is involved in embryonic development and carcinogenesis. We designed a novel anti-Cripto-1 artificial antibody and assessed the recognition to the antigen and the potential to suppress the growth of cancer stem cells. First, single chain antibody clones were isolated by bio-panning with the affinity to recombinant Cripto-1 protein from our original phage-display library. Then, the variable regions of heavy chain VH and light chain VL in each clone were fused to constant regions of heavy chain CH and light chain CL regions respectively. These fused genes were expressed in ExpiCHO-S cells to produce artificial humanized antibodies against Cripto-1. After evaluation of the expression levels, one clone was selected and the anti-Cripto-1 antibody was produced and purified. The purified antibody showed affinity to recombinant Cripto-1 at 1.1 pmol and immunoreactivity to cancer tissues and cell lines. The antibody was available to detect the immunoreactivity in tissue microarrays of malignant tumors as well as in Cripto-1 overexpressing cells. Simultaneously, the antibody exhibited the potential to suppress the growth of human colon cancer derived GEO cells overexpressing Cripto-1 with IC₅₀ at approximately 110 nM. The artificially humanized antibody is proposed to be a good candidate to target cancer cells overexpressing Cripto-1.     

Structural significance of the alkyl substituent at the C-4 of (+)-trans-verbenyl acetate in sex pheromonal activity of the American cockroach

The significance of the alkyl group at the C-4 of (+)-trans-verbenyl acetate, which is the sex pheromone mimic of the American cockroach, was investigated. Seven alcohols possessing an ethyl, propyl, or dimethyl group at this position of the 6,6-dimethylbicyclo[3.1.1]heptane skeleton were synthesized and evaluated by behavioral assay. All of the alcohols were inactive, while three of four acetates of the 2-alcohols induced sexual behavior in male cockroaches at the 0.02 or 0.5 mg dosage level, either of which is many orders of magnitude higher than the threshold level of the natural sex pheromones (10^–8 mg). Among the acetates, the compounds with a methyl group or an -oriented ethyl group at C-4 showed the highest activity. The results are discussed in terms of spatial requirements of the molecules for interactions with the receptor.Studies on the sex pheromone mimic of the American cockroach, (+)-trans-verbenyl acetate. Part VIII. For Part VII, seeComp. Biochem. Physiol.,70A: 229–234 (1981).     

Preparation of poly(p-phenylene terephthalamide)/poly(vinyl chloride) molecular composite and its thermal and mechanical properties

Poly(p-phenylene terephthalamide) (PPTA) was blended with poly(vinyl chloride) (PVC) by solution-blending method. PPTA was metalated for dissolving in dimethyl sulfoxide. Dimethyl sulfoxide was used as a common solvent. In PPTA/PVC composite, PPTA accelerated the thermal degradation of PVC. PPTA molecules are aggregated as microfibrillar form in PVC matrix. Such microfibrils are dispersed homogeneously in PVC matrix, according to polarizing microscopic observation. The average diameter of the microfibrils becomes smaller in the composite with lower content of PPTA. In the surface region of PPTA microfibrils the intermolecular hydrogen bonds between C? Cl of PVC and N? H of PPTA are formed. Young's modulus and the yield stress at room temperature were higher in the composites than those in PVC. The modulus of the composites was higher, especially at the high temperatures above their glass transition temperatures, than that in PVC. The temperature dependence of modulus can be calculated by using the mechanical model equivalent to the quasi-3-dimensional microfibrillar model which will be approximately applied to the composite structure. It becomes apparent that the modulus of the PPTA microfibrils evaluated by using the mechanical model is higher in the higher molecular weight PPTA.     

Mapping evapotranspiration using MODIS and MM5 Four-Dimensional Data Assimilation

Evapotranspiration (ET), the sum of evaporation from soil and transpiration from vegetation, is of vital importance in the hydrologic cycle and must be taken into consideration in assessments of the water resources of any region. The MODerate resolution Imaging Spectroradiometer (MODIS) sensor offers a promising opportunity for estimating daily ET with a 1 km spatial resolution, but is hampered by frequent cloud contamination or data gaps from other factors. In this study, 1) a stand-alone ET model was applied and tested during clear or partial cloudy sky conditions using MODIS-based inputs of land surface and atmospheric data and 2) meteorological simulations by using Four-Dimensional Data Assimilation (FDDA) system between MODIS and the 5th Generation Meso-scale Meteorological Model (MM5) was used in cloudy conditions to facilitate continuous daily ET estimates. The MODIS ET algorithm modified from Mu et al. (2007) is based on the Penman-Monteith equation and was applied to predict ET at flux measurement sites. This algorithm considers both the effects of surface energy partitioning processes and environmental constraints on ET. We devised gap-filling approaches for MODIS aerosol and albedo data that were identified as bottlenecks to determine retrieval rates of insolation and ET. MODIS-derived input variables (i.e., meteorological variables and radiation components) for estimating ET showed a good agreement with flux tower observations at each site. The retrieval rate of MODIS ET doubled at four flux measurement sites after gap-filling with negligible compensation was undertaken for accuracy. In spite of the high accuracy of MODIS-derived input variables, MODIS ET showed meaningful errors at the four flux measurement sites. These errors were mainly associated with errors in the estimated canopy conductance. During clear sky conditions, MODIS was used to calculate ET, while the MODIS-MM5 FDDA system provided input variables for the calculation of ET under cloudy sky conditions. The performance of the MODIS-MM5 FDDA system was evaluated by comparing ET based on MODIS, which showed a good agreement with the MODIS ET for various land cover types. Our results indicate that MODIS can be applied to monitor the land surface energy budget and ET with reasonable accuracy and that MODIS-MM5 FDDA has the potential to provide reasonable input data of ET estimation under cloudy conditions.     

Two-stage nuclear demagnetization refrigerator

Two-stage nuclear demagnetization has been performed using PrCu₆ and Cu as coolants. The Cu nuclear stage reached temperatures as low as 10 K with a rate of temperature rise of less than a few K/h. The corresponding conduction electron temperature is estimated to be less than 50 K. A nuclear orientation thermometer of Al ⁵⁴Mn has been successfully used to measure temperatures down to 10 K.Supported by the Japan Society for the Promotion of Science.