Proteomic Insights into Sulfur Metabolism in the Hydrogen-Producing Hyperthermophilic Archaeon Thermococcus onnurineus NA1

The hyperthermophilic archaeon Thermococcus onnurineus NA1 has been shown to produce H₂ when using CO, formate, or starch as a growth substrate. This strain can also utilize elemental sulfur as a terminal electron acceptor for heterotrophic growth. To gain insight into sulfur metabolism, the proteome of T. onnurineus NA1 cells grown under sulfur culture conditions was quantified and compared with those grown under H₂-evolving substrate culture conditions. Using label-free nano-UPLC-MS^E-based comparative proteomic analysis, approximately 38.4% of the total identified proteome (589 proteins) was found to be significantly up-regulated (≥1.5-fold) under sulfur culture conditions. Many of these proteins were functionally associated with carbon fixation, Fe–S cluster biogenesis, ATP synthesis, sulfur reduction, protein glycosylation, protein translocation, and formate oxidation. Based on the abundances of the identified proteins in this and other genomic studies, the pathways associated with reductive sulfur metabolism, H₂-metabolism, and oxidative stress defense were proposed. The results also revealed markedly lower expression levels of enzymes involved in the sulfur assimilation pathway, as well as cysteine desulfurase, under sulfur culture condition. The present results provide the first global atlas of proteome changes triggered by sulfur, and may facilitate an understanding of how hyperthermophilic archaea adapt to sulfur-rich, extreme environments.     

Effect of adding Ce on interfacial reactions between Sn–Ag solder and Cu

We investigated the effect of adding cerium (Ce) to low Ag content Sn–1.0wt.%Ag solder on the interfacial reactions between the Sn–1.0Ag solder and Cu substrate. The formation and growth of interfacial intermetallic compounds (IMCs) between the Sn–1.0Ag–0.3Ce solder and Cu substrate were studied and the results were compared to those obtained for the Ce-free Sn–1.0Ag/Cu and most promising Sn–3.0Ag–0.5Cu/Cu systems. The addition of Ce to the Sn–Ag solder significantly reduced the growth of the interfacial Cu–Sn IMCs, retarded the interfacial reactions between the solder and the substrate, and prevented the IMC from spalling from the interface. The Sn–1.0Ag–0.3Ce solder alloy had a good interfacial stability with the Cu substrate during solid-state isothermal aging in the viewpoint of IMC growth.     

Long-Lasting Growth Hormone Regulated by the Ubiquitin-Proteasome System

To increase the half-life of growth hormones, we proposed its long-lasting regulation through the ubiquitin-proteasome system (UPS). We identified lysine residues (K67, K141, and K166) that are involved in the ubiquitination of human growth hormone (hGH) using ubiquitination site prediction programs to validate the ubiquitination sites, and then substituted these lysine residues with arginine residues. We identified the most effective substituent (K141R) to prevent ubiquitination and named it AUT-hGH. hGH was expressed and purified in the form of hGH-His, and ubiquitination was first verified at sites containing K141 in the blood stream. Through the study, we propose that AUT-hGH with an increased half-life could be used as a long-lasting hGH in the blood stream.     

Effects of surface functional groups on film formation and tensile strength development in reactive latex systems

The effect of surface functional groups on chain interdiffusion at the interfacial zone of reactive latexes was investigated. A series of latexes with different surface functionalities was prepared by batch or shot-growth emulsion polymerizations. The particle surface properties were varied by changing the number density of functional groups. The rate of tensile strength development decreases with the increasing number density of surface functional groups, which indicated that inter-particle crosslinking restricted the interdiffusion of polymer chains during film formation and annealing. The average diffusion coefficient D of polymer chains across particle interfaces was obtained from dynamic mechanical analysis and compared well with the results from the rate of tensile strength development. The magnitude of D of the reactive latex film was lower than that of the homopolymer film. The lower chain mobility of reactive latexes prevented mechanical strength development.     

Characterization of prolyl oligopeptidase from hyperthermophilic archaeon Thermococcus sp. NA1

The prolyl oligopeptidase TNA1_POP was found to be encoded in the genome of the hyperthermophilic archaeon Thermococcus sp. NA1 and showed high similarities to its archaeal homologs (76-83%). The enzyme was found to be a single polypeptide composed of 616 amino acids with conserved signature domains. A recombinant TNA1_POP expressed in Escherichia coli was capable of hydrolyzing succinyl-Ala-Pro-p-nitroanilide (Suc-Ala-Pro-pNA) with temperature and pH optimums of 80 degrees C and 7, respectively. TNA1_POP activity appeared to be significantly activated by pre-incubation at 80 degrees C and 90 degrees C with the optimum temperature unchanged. The heat-activated enzyme exhibited a k(cat) approximately twofold higher than that of the unheated enzyme, however, both enzymes showed the same K(m). TNA1_POP was thermostable at 80 degrees C retaining 80% of its heat-activated activity even after 23 h, but it lost its enzymatic activity at 90 degrees C with a half-life of 3 h. The loss of the enzymatic activity at 90 degrees C seemed to be caused by the autodegradation of the enzyme, not by thermal denaturation, as supported by circular dichroism spectropolarimetry. Autodegradation fragments ranging from 2 to 18 kDa were mapped by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.     

Comparison of YAG: Eu phosphor synthesized by supercritical water and solid-state methods in a batch reactor

Phosphor yttrium aluminum garnet (Y₃Al₅O₁₂, YAG) doped with Eu (10 at%) was synthesized by supercritical water (SCW) and solid-state methods in a batch reactor. The crystals, morphologies and luminescent properties of the phosphors obtained from different pH conditions in the SCW were studied. Pure YAG phase was successfully synthesized at the alkaline condition in the SCW without formation of intermediate phases, while the size and luminescent property of YAG: Eu were strongly affected by pH condition. The luminescent intensity of YAG: Eu by the SCW method without further thermal treatment is stronger than that by the solid-state method with thermal treatment.     

Development of head detection and tracking systems for visual surveillance

This paper proposes a technique for the detection of head nod and shake gestures based on eye tracking and head motion decision. The eye tracking step is divided into face detection and eye location. Here, we apply a motion segmentation algorithm that examines differences in moving people’s faces. This system utilizes a Hidden Markov Model-based head detection module that carries out complete detection in the input images, followed by the eye tracking module that refines the search based on a candidate list provided by the preprocessing module. The novelty of this paper is derived from differences in real-time input images, preprocessing to remove noises (morphological operators and so on), detecting edge lines and restoration, finding the face area, and cutting the head candidate. Moreover, we adopt a K-means algorithm for finding the head region. Real-time eye tracking extracts the location of eyes from the detected face region and is performed at close to a pair of eyes. After eye tracking, the coordinates of the detected eyes are transformed into a normalized vector of x-coordinate and y-coordinate. Head nod and shake detector uses three hidden Markov models (HMMs). HMM representation of the head detection can estimate the underlying HMM states from a sequence of face images. Head nod and shake can be detected by three HMMs that are adapted by a directional vector. The directional vector represents the direction of the head movement. The vector is HMMs for determining neutral as well as head nod and shake. These techniques are implemented on images, and notable success is notified.     

Influence of phosphorous addition on Bi<Subscript>3</Subscript>Mo<Subscript>2</Subscript>Fe<Subscript>1</Subscript> oxide catalysts for the oxidative dehydrogenation of 1-butene

Bi₃Mo₂Fe₁P _x oxide catalysts were prepared by a co-precipitation method and the influence of phosphorous content on the catalytic performance in the oxidative dehydrogenation of 1-butene was investigated. The addition of phosphorous up to 0.4mole ratio to Bi₃Mo₂Fe₁ oxide catalyst led to an increase in the catalytic performance; however, a higher phosphorous content (above P=0.4) led to a decrease of conversion. Of the tested catalysts, Bi₃Mo₂Fe₁P_0.4 oxide catalyst exhibited the highest catalytic performance. Characterization results showed that the catalytic performance was related to the quantity of a π-allylic intermediate, facile desorption behavior of adsorbed intermediates and ability for re-oxidation of catalysts.     

Biochemical monitoring of black raspberry (Rubus coreanus Miquel) fruits according to maturation stage by H NMR using multiple solvent systems

Nuclear magnetic resonance (NMR) techniques coupled with multivariate data analysis were used to conduct monitoring of biochemical changes of black raspberry fruits at different stages of maturation and under various extraction and NMR dissolution solvent conditions: extraction with 50% methanol and D₂O as an NMR dissolution solvent, extraction with 50% methanol and 50% methanol-d₄ as an NMR dissolution solvent, and extraction with 100% ethyl acetate and 100% methanol-d₄ as an NMR dissolution solvent. Partial least-squares discriminant analysis reliably distinguished black raspberry fruits according to the maturation stage, whereby the relative levels of various compounds such as amino acids, organic acids, sugars and phenolic compounds were compared using analysis of variance. Sucrose and most of the amino acids, and organic acids decreased, whereas fructose, glucose, and cyanidins increased in relative concentration according to maturation of black raspberry fruits. The total number and kinds of assigned compounds of the three solvent systems were also compared. This research demonstrates that the metabolic profile of black raspberry fruits changes during maturation, and provides objective criteria for determining the stage of black raspberry fruit maturation via a ¹H NMR-based metabolomics technique using multiple solvent systems.     

Effect of Ca doping on the electrical conductivity of the high temperature proton conductor LaNbO4

The sintering properties, crystal structure and electrical conductivity of La_1−xCa_xNbO_4−δ (x = 0, 0.005, 0.01, 0.015, 0.02 and 0.025), prepared by a solid-state reaction, have been investigated using powder X-ray diffraction (XRD), scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDS), and electrochemical impedance spectroscopy (EIS). In 2.5% Ca-doped samples, a small amount of impurities Ca₂Nb₂O₇ were observed from the XRD patterns. Impedance spectra show that the grain boundary resistance increases with increasing Ca content, while the bulk resistance remains essentially constant below 550 °C. Despite the higher degree of grain growth observed for higher Ca doping levels, the total conductivity of the La_1−xCa_xNbO_4−δ series decreases with increasing Ca content from 0.5 to 2.0 mol%. The activation energy for the total conductivity decreases with increasing Ca content from 0.71 eV (x = 0) to 0.54 eV (x = 0.01) for the high temperature tetragonal phase, then it increases to 0.60 eV for x = 0.02. For the monoclinic phase, the activation energy exhibits similar trend except La_0.995Ca_0.005NbO_4−δ shows the lowest value of 1.26 eV. The Ca and Nb content present at the grain boundaries for La_0.99Ca_0.01NbO_4−δ are much higher than that on the grain surface, as determined from the EDS analysis. These results imply that the solubility of CaO in LaNbO₄ is in the range from 0.5 to 1.0 mol%. By increasing the sintering temperature from 1500 °C to 1550 °C, the proton conductivity of the Ca-doped LaNbO₄ was improved with enlarged grain size due to a reduction in the resistive grain boundary contribution.