Higher expression of topoisomerase II in lung cancers than normal lung tissues: different expression pattern from topoisomerase I

To examine the expression of topoisomerase I and topoisomerase II in primary lung cancer specimens at mRNA level, we carried out Northern blot analysis. As for topoisomerase I expression, there was no remarkable difference between lung cancer specimens and non-cancerous lung tissues. On the other hand, we could detect topoisomerase II mRNA in almost all lung cancer specimens, but not in non-cancerous tissues. By Southern blot analysis, we could not detect large deletion nor rearrangement in DNA level. These results suggest that the expression of topoisomerase II is highly increased in lung cancer at mRNA level and drugs against topoisomerase II might be more tumor-specific than those against topoisomerase I.     

Purification and characterization of a dehydrogenase catalyzing conversion of N alpha-benzyloxycarbonyl-L-aminoadipic-delta-semialdehyde to N alpha-benzyloxycarbonyl-L-aminoadipic acid from rhodococcus sp. AIU Z-35-1

The enzyme catalyzing conversion of N alpha-benzyloxycarbonyl-L-aminoadipic-delta-semialdehyde (N alpha-Z-L-AASA) to N alpha-benzyloxycarbonyl-L-aminoadipic acid (N alpha-Z-L-AAA) in Rhodococcus sp. AIU Z-35-1 was identified, and its characteristics were revealed. This reaction was catalyzed by a dehydrogenase with a molecular mass of 59 kDa. The dehydrogenase exhibited enzyme activity on not only N alpha-Z-L-AASA but also N alpha-Z-D-AASA and short chain aliphatic aldehydes, but not on aromatic aldehydes and alcohols. The apparent Km values for N alpha-Z-L-AASA, N alpha-Z-D-AASA and NAD+ were estimated to be 3.8 mM, 14.1 mM and 0.16 mM, respectively. The NH2 terminal amino acid sequence of this enzyme exhibited a similarity to those of a piperidein-6-carboxylate dehydrogenase from Streptomyces clavuligerus and a putative dehydrogenase from Rhodococcus sp. RHA 1, but not to those of other microbial aldehyde dehydrogenases.     

Lactoperoxidase activity in milk is correlated with somatic cell count in dairy cows

Lactoperoxidase (LPO) is a milk protein with antimicrobial function. The present study was undertaken to examine the correlation between LPO activity and somatic cell count (SCC) in milk to use LPO activity as an indicator of mastitis. Composite milk of 36 cows and quarter milk of 3 cows were collected once per week from 0 to 300 d postpartum and twice per day for 1 wk, respectively. For the measurement of LPO activity, milk was mixed with tetramethylbenzidine solution and incubated at 37°C for 30 min, followed by the measurement of optical density. When only milk with low SCC (132 ± 12 × 10³ cells/mL) was used, a significant decrease in LPO activity was detected in primiparous cows from 0 to 4 mo postpartum. Lactoperoxidase activities of primiparous cows in mo 1, 2, and 3 postpartum were significantly higher than those in multiparous cows. When composite milk was divided based on LPO activity, the SCC was significantly higher in the groups with LPO activity >5 and from 3 to 3.9 U/mL in the second- and fourth-parity cows, respectively, compared with the group with LPO activity <2 U/mL. Extremely high SCC were found in the ≥fifth-parity cows, even in low-LPO activity groups. In the case of quarter milk, higher LPO activity was associated with increased SCC in all 3 cows. The percentage of quarter milk samples with high SCC (4,062 ± 415 × 10³ cells/mL) increased with an increase in the LPO activity. The percentage of quarter milk samples with high SCC was 50.0 to 100% in the milk with LPO activity ≥5 U/mL. These results indicate that the correlation of LPO activity to the SCC in bovine milk may point to the potential use of the former as an indicator of SCC.     

System for augmented human–robot interaction through mixed reality and robot training by non-experts in customer service environments

Human–robot interaction during general service tasks in home or retail environment has been proven challenging, partly because (1) robots lack high-level context-based cognition and (2) humans cannot intuit the perception state of robots as they can for other humans. To solve these two problems, we present a complete robot system that has been given the highest evaluation score at the Customer Interaction Task of the Future Convenience Store Challenge at the World Robot Summit 2018, which implements several key technologies: (1) a hierarchical spatial concepts formation for general robot task planning and (2) a mixed reality interface to enable users to intuitively visualize the current state of the robot perception and naturally interact with it. The results obtained during the competition indicate that the proposed system allows both non-expert operators and end users to achieve human–robot interactions in customer service environments. Furthermore, we describe a detailed scenario including employee operation and customer interaction which serves as a set of requirements for service robots and a road map for development. The system integration and task scenario described in this paper should be helpful for groups facing customer interaction challenges and looking for a successfully deployed base to build on.     

Structural interlock between ligand-binding site and stalk-like region of beta1 integrin revealed by a monoclonal antibody recognizing conformation-dependent epitope

Integrin activation and sebsequent ligand binding to it are regulated by intracellular mechanisms called inside-out signaling, which are not fully understood and are accompanied by dynamic structural changes of the integrin molecule itself. A monoclonal antibody recognizing a conformation-dependent epitope on human beta1 integrin was produced and characterized in detail. This antibody, AG89, reacted with human integrin beta1 chain regardless of the alpha subunit. AG89 can recognize resting state beta1 integrin on the cells, but the reactivity is increased approximately 2-fold upon integrin activation by activating anti-beta1 antibodies and approximately 3-fold by Mn2+. Furthermore, occupation of the ligand-binding pocket by a soluble ligand (RGD peptide for alpha(v)beta1 and CS-1 peptide for alpha4beta1) resulted in maximum binding of AG89, indicating that the epitope for AG89 is exposed during the conformational changes of beta1 integrin upon activation/ligation. Epitope mapping by using interspecies chimeric beta1 revealed that the epitope for AG89 lies within residues 426-587, which corresponds to the cysteine-rich repeat structure located in the middle of the beta1 chain. The fact that binding of AG89 itself could activate the resting beta1 integrin indicates that exposure of the AG89 epitope in the membrane-proximal stalk-like domain and "opening" of the ligand-binding pocket at the outermost domain are physically linked. We propose that the integrin "signaling" is mediated by this direct physical transduction of conformational information along the integrin molecule.     

Imaging the passage of a single hydrocarbon chain through a nanopore

Molecular transport through nanoscale pores in films, membranes and wall structures is of fundamental importance in a number of physical, chemical and biological processes. However, there is a lack of experimental methods that can obtain information on the structure and orientation of the molecules as they pass through the pore, and their interactions with the pore during passage. Imaging with a transmission electron microscope is a powerful method for studying structural changes in single molecules as they move and for imaging molecules confined inside carbon nanotubes. Here, we report that such imaging can be used to observe the structure and orientation of a hydrocarbon chain as it passes through nanoscale defects in the walls of a single-walled carbon nanotube to the vacuum outside, and also to study the interactions between the chain and the nanopore. Based on experiments at 293 K and 4 K we conclude that the major energy source for the molecular motions observed at 4 K is the electron beam used for the imaging.     

Effect of dispersant on paste rheology in preparation of porous alumina with oriented pores by extrusion method

The effect of additives on paste rheology was investigated for preparation of porous ceramics with unidirectionally aligned cylindrical pores. Ammonium poly-carboxylic acid (APA) used as a dispersant and it was adsorbed on alumina powder surface. The adsorption isotherm of APA was fitted by Langmuir equation. The saturated monolayer adsorption was 5.9 mg/g. The apparent viscosity became a minimum at 0.8 mass % of APA corresponding to 71.2 mPa⋅s. This APA amount of 5.6 mg/g, is in good agreement with the observed APA amount. Since the nylon 66 fibers (0–35 vol. %) mixed with the alumina powder have a strong interaction with each other, they became twisted and agglomerated. This agglomeration increased with increasing fiber content but decreased by adding oleic acid. The pastes with added oleic acid were capable of being extruded at higher pressure. The obtained porous alumina ceramics showed highly oriented cylindrical pores parallel to the extrusion direction. The pore orientation was higher in the oleic acid added pastes than those without oleic acid. The added nylon 66 fibers are mostly converted to pores while maintaining the original shape after sintering. The pore size distribution of the obtained porous ceramics measured by mercury porosimetry method showed a peak at about 4 μm which is apparently smaller than that observed in the SEM photographs and the obtained result is considered to be corresponding to the necks formed by fiber contacts.     

Structural characterization of photoreduced complete and incomplete cubane-type molybdenum oxide clusters in CO and their catalytic behavior in propene metathesis reaction

SiO2-impregnated complete and incomplete cubane-type molybdenum oxide clusters such as [(RhCp*)₄Mo₄O₁₆] (I) and [(RhCp*)₂Mo₃O₉(OMe)₄] (II) were photoreduced with a UHP-Hg lamp (> 365 nm) in CO, resulting in the formation of catalytically active species for propene metathesis at 300 K. The CO-photoreduced Mo oxide clusters were characterized by EXAFS, IR, XPS and TPD technique. The results suggest that the Mo-O_4c (four-centered bridging oxygens) in I were specifically reduced with CO under the illumination to produce two sets of subcarbonyl species, e.g., those characteristic of the IR bands at 2061 and 2021 cm^–1, which were thermally inactive for the¹³CO exchange reaction but very active under illumination at 300 K, possibly assignable to Mo(CO)x (x = 2, 3), whereas those at 2092 and 2035 cm^–1 due to Rh(CO)₂, which were readily exchangeable with¹³CO at 300 K in dark. Removal of both carbonyls attached on Rh and Mo in photoreduced I and II by evacuation at 375–440 K led to the formation of oxygen-deficient Mo⁴+/Mo⁵⁺ sites, which exhibited high catalytic activities in propene metathesis at 300 K to produce an equimolar mixture of ethene and 2-butenes. The CO-photoreduced incomplete cubane Mo oxide cluster (II) exhibited higher activities and higher trans/cis ratios of 2-butenes in the reaction, compared with those on the photoreduced I.