The Atmospheric Scanning Electron Microscope with open sample space observes dynamic phenomena in liquid or gas

Although conventional electron microscopy (EM) requires samples to be in vacuum, most chemical and physical reactions occur in liquid or gas. The Atmospheric Scanning Electron Microscope (ASEM) can observe dynamic phenomena in liquid or gas under atmospheric pressure in real time. An electron-permeable window made of pressure-resistant 100 nm-thick silicon nitride (SiN) film, set into the bottom of the open ASEM sample dish, allows an electron beam to be projected from underneath the sample. A detector positioned below captures backscattered electrons. Using the ASEM, we observed the radiation-induced self-organization process of particles, as well as phenomena accompanying volume change, including evaporation-induced crystallization. Using the electrochemical ASEM dish, we observed tree-like electrochemical depositions on the cathode. In silver nitrate solution, we observed silver depositions near the cathode forming incidental internal voids. The heated ASEM dish allowed observation of patterns of contrast in melting and solidifying solder. Finally, to demonstrate its applicability for monitoring and control of industrial processes, silver paste and solder paste were examined at high throughput. High resolution, imaging speed, flexibility, adaptability, and ease of use facilitate the observation of previously difficult-to-image phenomena, and make the ASEM applicable to various fields.     

Human colorectal carcinomas specifically accumulate Mr 42,000 ubiquitin-conjugated cytokeratin 8 fragments

Recent studies have shown that various tumor cells accumulate ubiquitin (Ub)-conjugated proteins, the profiles of which differ from those of normal cells. To identify the Ub-conjugated proteins accumulated specifically by human carcinoma cells, a two-dimensional immunoblot analysis of 31 surgically resected human primary colorectal carcinoma tissues was performed using an anti-Ub monoclonal antibody, KM691. Two distinct Mr 42,000 and 45,000 proteins in the Triton X-insoluble fractions of carcinoma tissues reacted with this antibody, whereas only one Mr 45,000 protein reacted in normal tissues. The Mr 42,000 Ub-conjugated proteins were specific to carcinoma tissues from 25 patients (80.6%). One of the purified Mr 42,000 proteins was digested with Achromobacter protease I. This protein was identified as a cytokeratin 8 (CK 8) fragment based on both molecular mass determination and molecular mass searching of Achromobacter protease I-digested fragments of proteins registered in a protein sequence data base. Two-dimensional immunoblot analysis with an anti-CK 8 antibody confirmed that all of the Mr 42,000 proteins were CK 8 degradation products. These results demonstrate that human colorectal carcinomas specifically accumulate Mr 42,000 Ub-conjugated CK 8 fragments. This accumulation was observed frequently not only in advanced (18/22, 81.8%), but also in early stage cases (7/9, 77.8%), suggesting that it occurs even in the early stages of colorectal carcinoma progression.     

Cloning of the gene for monogalactosyldiacylglycerol synthase and its evolutionary origin

Monogalactosyldiacylglycerol (MGDG) synthase (UDPgalactose:1,2-diacylglycerol 3-beta-D-galactosyltransferase; EC 2.4.1.46) catalyzes formation of MGDG, a major structural lipid of chloroplast. We cloned a cDNA for the synthase from cucumber cDNA library. The full-length cDNA clone was 2142 bp, and it contains a 1575-bp open reading frame encoding 525 aa. The open reading frame consists of the regions for a mature protein (422 aa; Mr of 46,552) and transit peptide to chloroplast (103 aa). Although the molecular weight of mature protein region matched that purified from cucumber cotyledons, it was quite different from those purified from spinach (approximately 20 kDa) reported by other groups. The mature region of the protein was expressed in Escherichia coli as a fusion protein with glutathione S-transferase. The expression in E. coli showed that the protein catalyzed MGDG synthesis very efficiently. Therefore, we concluded that the cDNA encodes MGDG synthase in cucumber. In addition, the deduced amino acid sequence of the MGDG synthase cDNA showed homology with MurG of Bacillus subtilis and E. coli, which encode a glycosyltransferase catalyzing the last step of peptidoglycan synthesis in bacteria. This sequence homology implies that the machinery of chloroplast membrane biosynthesis is evolutionarily derived from that of cell wall biosynthesis in bacteria. This is consistent with the endosymbiotic hypothesis of chloroplast formation.     

CAD-compatible high-speed CMOS/SIMOX gate array using field-shieldisolation

A specific 0.5 μm CMOS/SIMOX technology was developed for a gate array/sea of gate (SOG) using field-shield (FS) isolation to overcome a pending problem of source-to-drain breakdown voltage (BV_ds) lowering. FS isolation is capable of improving BV_ds because surplus holes generated by impact ionization at the drain region are collected through the body region under the FS gate. BV_ds was maintained at a level of junction breakdown before reaching the punchthrough limitation at a gate length of around 0.3 μm using the FS isolation. The FS isolation technique was successfully applied to an SOG gate array on a SIMOX substrate. The gate array has the same area as that on the bulk-Si and is compatible to a conventional bulk-Si CAD system because the layout is basically the same. A 53-stage ring oscillator fabricated on the FS isolated SOG gate array exhibited 1.7 times higher speed operation than that on a bulk-Si counterpart, keeping low power consumption characteristics up to a drain voltage of 3 V     

Analysis of molecular dynamics of colloidal particles in transported dilute samples by self-mixing laser Doppler velocimetry

Colloidal particles in a liquid medium are transported with constant velocity, and dynamic light scattering experiments are performed on the samples by self-mixing laser Doppler velocimetry. The power spectrum of the modulated wave induced by the motion of the colloidal particles cannot be described by the well-known formula for flowing Brownian motion systems, i.e., a combination of Doppler shift, diffusion, and translation. Rather, the power spectrum was found to be described by the q-Gaussian distribution function. The molecular mechanism resulting in this anomalous line shape of the power spectrum is attributed to the anomalous molecular dynamics of colloidal particles in transported dilute samples, which satisfy a nonlinear Langevin equation.     

Novel cephalosporin derivatives possessing a bicyclic heterocycle at the 3-position. Part I: Synthesis and biological activities of 3-(benzothiazol-2-yl)thiocephalosporin derivatives, CP0467 and related compounds

Insertional mutagenesis in transgenic mice is a powerful method to study structure/function relationships between individual genes and complex developmental traits in the whole organism. Unlike spontaneous or chemical-induced mutations, insertional mutations have the advantage that the mutant locus is "tagged" with the transgene and, therefore, readily accessible at the molecular level. Starting with the work on the limb deformity locus, we describe here the characterization of several mouse mutants generated by insertional mutagenesis with the pronuclear microinjection procedure. These transgenic lines have proven to be ideal as models for human disease and for studying the function of novel genes during development. We also describe the unique features of insertional mutations that arise in transgenic mice produced with the pronuclear microinjection procedure and provide recommendations on how to clone and characterize these mutations at the molecular level. Finally, we discuss future prospects for the use of this unique form of germline mutagenesis in the mouse.     

Modifications of a macrolide antibiotic midecamycin. II. Reaction of midecamycin and 9-acetylmidecamycin with dimethylsulfoxide and acetic anhydride

Treatment of 9,2'-diacetylmidecamycin (2) with DMSO and acetic anhydride afforded 3'-methylthiomethyl derivative (3) preferably in the presence of pyridine. Reaction of midecamycin (1) with DMSO and acetic anhydride gave 2'-acetyl-9-dehydro-3'-methylthiomethyl derivative (9) indicating that the three hydroxyl groups reacted in a different way to the reagent. When compound 2 was reacted with DMSO and acetic anhydride in the presence of CCl4, 3'-acetoxymethyl derivative (13) was a major product, which was formed via 3 through the Pummerer rearrangement. The structures of 3, 9 and 13 were confirmed by examining NMR and mass spectra of these compounds and their deuterio analogue. They showed antimicrobial spectra similar to 1 but superior in vivo activity.     

Silicon-on-sapphire m.o.s.f.e.t.s fabricated by back-surface laser-anneal technology

Self-aligned aluminium-gate silicon-on sapphire (s.o.s.) m.o.s.f.e.t.s have been fabricated by applying laser-anneal technology from the back surface for activation of the ion-implanted channel layer. The threshold voltage and surface electron mobility were similar to the conventional silicon-gate m.o.s.f.e.t.s, but the low resistivity of gate and interconnection electrodes using aluminium is advantageous for high switching speed m.o.s. l.s.i.     

A relation between calculated human body exergy consumption rate and subjectively assessed thermal sensation

Application of the exergy concept to research on the built environment is a relatively new approach. It helps to optimize climate conditioning systems so that they meet the requirements of sustainable building design. As the building should provide a healthy and comfortable environment for its occupants, it is reasonable to consider both the exergy flows in building and those within the human body.Until now, no data have been available on the relation between human-body exergy consumption rates and subjectively assessed thermal sensation. The objective of the present work was to relate thermal sensation data, from earlier thermal comfort studies, to calculated human-body exergy consumption rates.The results show that the minimum human body exergy consumption rate is associated with thermal sensation votes close to thermal neutrality, tending to the slightly cool side of thermal sensation.Generally, the relationship between air temperature and the exergy consumption rate, as a first approximation, shows an increasing trend. Taking account of both convective and radiative heat exchange between the human body and the surrounding environment by using the calculated operative temperature, exergy consumption rates increase as the operative temperature increases above 24 °C or decreases below 22 °C. With the data available so far, a second-order polynomial relationship between thermal sensation and the exergy consumption rate was established.     

Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase)

The small guanosine triphosphatase Rho is implicated in myosin light chain (MLC) phosphorylation, which results in contraction of smooth muscle and interaction of actin and myosin in nonmuscle cells. The guanosine triphosphate (GTP)-bound, active form of RhoA (GTP.RhoA) specifically interacted with the myosin-binding subunit (MBS) of myosin phosphatase, which regulates the extent of phosphorylation of MLC. Rho-associated kinase (Rho-kinase), which is activated by GTP.RhoA, phosphorylated MBS and consequently inactivated myosin phosphatase. Overexpression of RhoA or activated RhoA in NIH 3T3 cells increased phosphorylation of MBS and MLC. Thus, Rho appears to inhibit myosin phosphatase through the action of Rho-kinase.