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.     

Effects of short- and long-term dexamethasone treatment on growth and growth hormone (GH)-releasing hormone (GRH)-GH-insulin-like growth factor-I axis in conscious rats

Although the inhibitory effects of a chronic excess of glucocorticoids (GC) on body growth and GH secretion are well established, the mechanisms involved remain unclear. In this study, we examined the chronic effects of a high dose of dexamethasone (DEX) on spontaneous GH secretion and insulin-like growth factor (IGF)-I in conscious rats. The animals were given daily i.p. injections of DEX (200 microg/day) for either one or four weeks. Body growth assessed by tibia length and serum IGF-I levels was significantly inhibited 1 week after treatment. By contrast, spontaneous GH secretion was not altered 1 week after the treatment. Neither hypothalamic GRH and somtatostain mRNA levels nor GH responses to GRH from single somatotropes were affected 1 week after the treatment. Four weeks after DEX treatment, body growth of the rats was noticeably suppressed. Interestingly, spontaneous GH secretion, hypothalamic GRH mRNA levels and GH responses to GRH were all inhibited 4 weeks after treatment. Pituitary GRH receptor mRNA levels were not altered 1 week after treatment, but increased after 4 weeks. These results indicate that a high dose of DEX initially impairs IGF-I production and subsequently inhibits spontaneous GH secretion in rats. Inhibition of spontaneous GH secretion resulting from chronic GC excess is due, at least in part, to the impairment of hypothalamic GRH synthesis and pituitary GH responsiveness. An increase in the pituitary GRH receptor may be caused by decreased GRH secretion.     

Non-linear current–voltage characteristics related to native defects in SrTiO3 and ZnO bicrystals

SrTiO₃ and ZnO bicrystals with various types of boundaries were fabricated in order to examine their current–voltage characteristics across single grain boundaries. Their grain boundary structures were also investigated by high-resolution transmission electron microscopy. In Nb-doped SrTiO₃, electron transport behaviors depend on the type of boundaries. Random type boundaries exhibit highly non-linear current–voltage characteristics, while low angle boundaries show a slight non-linearity. On the contrary, undoped ZnO does not exhibit non-linear current–voltage characteristics in any type of boundaries including random ones. It is suggested that the differences observed in current–voltage properties between the two systems are mainly due to the difference in the accumulation behavior of acceptor-like native defects at grain boundaries. A clear non-linearity is obtained by means of Co-doping even for the highly coherent Σ1 boundary in a ZnO bicrystal. This is considered to result from the production of acceptor-like native defects by Co-doping.     

Adsorption of urea,creatinine, and uric acid from three solution types using spherical activated carbon and its recyclability

In this paper, we propose that the urinary toxins from the wastewater be adsorbed on an adsorbent such as spherical activated carbon and the latter be regenerated by subjecting it to high temperatures to recycle activated carbon and also to recycle the water used in dialysis. We studied the adsorption of artificial waste dialysate, which is a mixed solution of urea, creatinine, and uric acid, and the separate solutions for each of these and found that their extents of adsorption onto the spherical activated carbon material were nearly identical. The amount of adsorption was approximately 1.4 mg·g^-1 for urea, 18 mg·g^-1 for creatinine, and 20 mg·g^-1 for uric acid. The urea, creatinine, and uric acid adsorbed onto the spherical activated carbon decomposed on heat treatment at 500℃, and the adsorption capacity of the spherical activated carbon was regenerated. Our study successfully demonstrated that the spherical activated carbon can be recycled in the waste dialysate treatment process.     

Mechanical strengthening of Si cantilever by chemical KOH etching and its surface analysis by TEM and AFM

Mechanical strengthening of a Si cantilever by applying KOH wet etching was investigated. Two kinds of Si cantilever specimens having the different crystallographic orientations of the sidewall surfaces, i.e., Si{100} and Si{110}, were fabricated from the same SOI wafer by a Bosch process. The typical height and pitch of the scalloping formed on the sidewall were 248 and 917 nm, respectively. A 50 % KOH (40 °C) chemical wet etching was applied to increase the fracture stress of the Si cantilever. The fracture stress in the both of Si{100} and Si{110} cantilevers increased with the advance of the etching. The obtained maximum fracture stress in Si{100} and Si{110} were 4.2 and 3.7 GPa, respectively. Sidewall surface of the cantilever was analyzed to investigate the mechanical strengthening of Si cantilever by wet etching. The etched surface crystalline was analyzed by the transmission electron microscope (TEM), and confirmed that the thickness of the affected flow layer was less than 10 nm from the obtained TEM image. Then the change of the surface roughness by the KOH etching was analyzed by the atomic force microscope. The surface was smoothened with the advance of the KOH etching. The roughness value of Ra in Si{100} and Si{110} decreased to 12.1 and 37.7 nm, respectively.     

Experimental measurement on tangential momentum accommodation coefficient in a single microtube

The tangential momentum accommodation coefficient (TMAC) was investigated experimentally from the mass flow rate through a single microtube under the slip flow and the early part of the transition regime. The measurements were carried out by the constant-volume method under the mean Knudsen number smaller than 0.3, which is based on the mean pressure of the inlet and the outlet of the microtube, to apply the second-order slip boundary condition. To measure TMACs on various materials, quite large microtube was employed, which require the reduction in leakage. TMAC was obtained from the slip coefficient determined by the relation of the mass flow rate to the mean Knudsen number. The obtained mass flow rate was well explained by the theoretical equation. TMACs of deactivated-fused silica with argon, nitrogen, and oxygen were measured, showing the tangential momentum was not accommodated completely to the surface, and the values showed good agreement with previous studies. From the comparison between microtubes with different inner diameter, it is showed that TMAC is determined mainly by gas species and surface material.     

Effect of the dilation caused by helium bubbles on edge dislocation motion in α-iron: molecular dynamics simulation

Various types of nanometric defects such as voids and helium (He) bubbles produced by high-energy neutron irradiations are known to degrade the mechanical properties of irradiated materials. In this study, we have evaluated the obstacle strength of He bubbles to the mobility of an edge dislocation in α-iron for 2 and 4 nm bubbles with He-to-vacancy (He/V) ratios ranging from 0 to 1 at 300 and 500 K, by molecular dynamics simulation. Results showed that as the He/V ratio increases, the obstacle strength needed for the release of a dislocation from the bubble becomes stronger up to a moderate He/V ratio (0.6 and 0.4 for 2 and 4 nm bubbles, respectively, at both temperatures), and a further increase in the He/V ratio leads to weakening of the obstacle strength. For He/V = 1, the obstacle strengths are 10–30% weaker than those at moderate He/V ratios depending on the bubble size and temperature. The extent of obstacle strength was found to be correlated with the dilation caused by He bubbles depending on the bubble size, He/V ratio, and temperature.     

Methylcyclohexane dehydrogenation for hydrogen production via a bimodal catalytic membrane reactor

The dehydrogenation of methylcyclohexane (MCH) to toluene (TOL) for hydrogen production was theoretically and experimentally investigated in a bimodal catalytic membrane reactor (CMR), that combined Pt/Al₂O₃ catalysts with a hydrogen‐selective organosilica membrane prepared via sol‐gel processing using bis(triethoxysilyl) ethane (BTESE). Effects of operating conditions on the membrane reactor performance were systematically investigated, and the experimental results were in good agreement with those calculated by a simulation model with a fitted catalyst loading. With H₂ extraction from the reaction stream to the permeate stream, MCH conversion at 250°C was significantly increased beyond the equilibrium conversion of 0.44–0.86. Because of the high H₂ selectivity and permeance of BTESE‐derived membranes, a H₂ flow with purity higher than 99.8% was obtained in the permeate stream, and the H₂ recovery ratio reached 0.99 in a pressurized reactor. A system that combined the CMR with a fixed‐bed prereactor was proposed for MCH dehydrogenation. © 2015 American Institute of Chemical Engineers AIChE J, 61: 1628–1638, 2015