Family M42 aminopeptidase from the syntrophic bacterium Symbiobacterium thermophilum: characterization using recombinant protein

The chromosomal DNA of the syntrophic thermophile Symbiobacterium thermophilum contains open reading frames of the genes encoding family M42 aminopeptidases, Pep1079, Pep1080, and Pep1081. To characterize these peptidases, the genes were cloned into Escherichia coli and overexpressed. Our experiments using the recombinant proteins confirmed that Pep1079, Pep1080, and Pep1081 are components of arginyl or lysinyl aminopeptidases that require Co2+ for enzymatic activity. Coexistence of Pep1079 and Pep1080 is necessary for expressing high peptidase activity. Pep1081 enhances the activity of Pep1079 and Pep1080.     

Genotoxicity of nano/microparticles in in vitro micronuclei, in vivo comet and mutation assay systems

Background

Recently, manufactured nano/microparticles such as fullerenes (C₆₀), carbon black (CB) and ceramic fiber are being widely used because of their desirable properties in industrial, medical and cosmetic fields. However, there are few data on these particles in mammalian mutagenesis and carcinogenesis. To examine genotoxic effects by C₆₀, CB and kaolin, an in vitro micronuclei (MN) test was conducted with human lung cancer cell line, A549 cells. In addition, DNA damage and mutations were analyzed by in vivo assay systems using male C57BL/6J or gpt delta transgenic mice which were intratracheally instilled with single or multiple doses of 0.2 mg per animal of particles.

Results

In in vitro genotoxic analysis, increased MN frequencies were observed in A549 cells treated with C₆₀, CB and kaolin in a dose-dependent manner. These three nano/microparticles also induced DNA damage in the lungs of C57BL/6J mice measured by comet assay. Moreover, single or multiple instillations of C₆₀ and kaolin, increased either or both of gpt and Spi^- mutant frequencies in the lungs of gpt delta transgenic mice. Mutation spectra analysis showed transversions were predominant, and more than 60% of the base substitutions occurred at G:C base pairs in the gpt genes. The G:C to C:G transversion was commonly increased by these particle instillations.

Conclusion

Manufactured nano/microparticles, CB, C₆₀ and kaolin, were shown to be genotoxic in in vitro and in vivo assay systems.     

Fabrication of damascene Cu wirings using solid acidic catalyst

The copper damascene process is one of the most promising technologies for fabricating Cu wirings for electronic devices such as LSIs. In this research, the fabrication of damascene Cu wirings was conducted using solid acidic catalyst. When a Cu-plated wafer, whose oxide is a basic oxide is dipped into a mixture of oxidizing solution and acidic solution, surface atoms are ionized and etched off into the solution. However, because conventional nonelectrolytic etching does not have a reference surface, it is difficult to utilize for planarization. Therefore, a new nonelectrolytic machining method using a cation-exchange fabric instead of an acidic solution was developed. To be more precise, the planarization of a Cu-plated wafer was carried out by rubbing with the cation-exchange fabric in ozone water. Basically, this method exploits chemical reactions so that the physical properties of the workpiece surface are not deteriorated. Furthermore, this method uses no chemicals except for ozone water, which easily dissociates into water and oxygen molecules; thus, this method is a low-cost, environmentally friendly process. In this paper, as a preliminary experiment, the nonelectrolytic etching of a Cu sample using solutions of O₃ and CO₂ was carried out to inspect the dependence of the etching rate on [O₃] and [H⁺]. The results indicate that the etching rate increased as [O₃] and [H⁺] increased. When [H⁺] was high relative to [O₃], a smooth etch-pit-free surface was achieved. Next, nonelectrolytic etching using a cation-exchange fabric was carried out, and properties similar to those in the case of etching using solutions were obtained. Finally, damascene Cu wirings were fabricated using ozone water and a cation-exchange catalyst.     

Fluticasone propionate reduced the production of GM-CSF, IL-6 and IL-8 generated from cultured nasal epithelial cells

The stages of mental rotation of complicated three-dimensional figures were studied using the intracortical interaction mapping. The role of the parietal areas in the mental rotation was shown. The frontal, central and the right temporal areas are involved in the prerotation setup. Both frontal areas and the left temporo-parieto-occipital areas participate in decision making and verbal response. In the case of unsuccessful task solving the process stops at the prerotation setup stage. During the verbal control the zones of the predominant connections at the first 3 stages are close to those in the background activity.     

Generation of 80-nm Wavelength-Tunable 100-fs Pulse Based on Comblike Profiled Fiber Comprised of HNLF and Zero Dispersion-Slope NZDSF

To realize extremely wideband wavelength-tunable femtosecond (fs) pulse generation, we propose a dispersion-flattened comblike profiled fiber that is a pulse compressor comprised of low dispersion-slope highly nonlinear fiber and zero dispersion-slope nonzero dispersion-shifted fiber (ZDS-NZDSF). We also apply ZDS-NZDSF to a soliton converter to reshape and compress a seed pulse train generated from a tunable laser source and an intensity modulator. Using the proposed scheme, we successfully generate 100-fs optical pulse from the seed pulse with the varied wavelengths of 1530, 1570, and 1610 nm     

Carinal resection for primary lung cancer--with special attention to a modified double-barrel method

Carinal resection for primary lung cancer was clinically evaluated. Carinal resection was performed in 18 patients, 17 males and one female, with a mean age of 64 years. Nine patients underwent carinal reconstruction and the other 9 sleeve or wedge pneumonectomy. The carinal reconstruction was of the montage type in one patient, the one-stoma type in 2, and the modified double-barrel method in 6. The modified double-barrel method is a technique that we developed by adding bronchial end-to-side anastomosis to the tracheobronchial end-to-end anastomotic site. A pedicled intercostal muscle flap was used for covering the anastomotic site. The postoperative respiratory complications after carinal reconstruction were anastomosis failure in 4 patients (pin-hole in 3) and respiratory failure in 2. However, no anastomosis stricture occurred, and recovery was satisfactory. There were no respiratory complications after pneumonectomy. One patient had renal failure before surgery and died of multiple organ failure 23 days after a montage type carinal reconstruction. The other 17 patients did well and could be discharged from the hospital and the overall mortality rate was 5.6%. No anastomosis stricture occurred in the modified double-barrel method. By carinal reconstruction covering of the anastomotic site is mandatory to prevent fatal postoperative complications.     

Genetic organization of GBV-C]

The chimaeric molecule rscu-PA-40 kDA/Hir (M23) comprises the kringle and protease domain of saruplase (rscu-PA) and a thrombin inhibitory domain fused to the C-terminus of the protease domain. The 27 amino cid long thrombin inhibitory domain contains a sequence directed to the active site of thrombin and a fragment from the C-terminal region of hirudin. 125I-radiolabelled M23 (0.03 microM) bound to thrombin that was immobilised onto CNBr-activated sepharose beads. Unlabelled M23 (0.01-10 microM) and hirudin (0.001-10 microM) concentration-dependently displaced 125I-M23 from its binding to thrombin. Saruplase (up to 10 microM) did not influence the thrombin binding of M23. The fibrinolytic properties of M23 and saruplase were compared in anaesthetized dogs with femoral artery and saphenous vein thrombosis. Under concomitant heparinization, the intravenous bolus injections of 1 mg/kg M23 or saruplase induced reperfusion of thrombotically occluded femoral arteries in 4 out of 5 treated animals in each case. There was one reocclusion in the M23-treated group. Time to reperfusion (23 +/- 4 vs 25 +/- 11 min) and maximal height of reperfusion blood flow (98 +/- 21 vs 108 +/- 15% of baseline flow) did not differ significantly between the treatment groups. The time course of the lysis of incorporated 125I-fibrin radioactivity in thrombosed saphenous-veins was similar after bolus injections of M23 and saruplase. The maximal dissolution of 125I-fibrin in the venous thrombosis model was 91 +/- 1% in M23- and 88 +/- 5% in saruplase-treated animals. Plasma levels of fibrinogen were not influenced and alpha 2-antiplasmin levels were slightly reduced (-27 +/- 3%) after bolus injection of M23. In contrast, bolus injection of saruplase was accompanied by a significant decrease of fibrinogen (-55 +/- 19%) and alpha 2-antiplasmin (-75 +/- 11%) plasma levels. Template bleeding times virtually did not differ before (2.8 +/- 0.3 min) and 60 min after bolus injection of M23 (3.1 +/- 0.3 min), whereas treatment with saruplase resulted in a significant prolongation of template bleeding time from 2.6 +/- 0.2 min to 28 +/- 13 min. It is concluded that the saruplase derivative M23, while inducing equieffective thrombolysis after intravenous bolus injection in dogs, causes much fewer haemostatic side effects than its parent molecule. The high thrombus-specific activity of M23 is tentatively attributed to its affinity to clot-bound thrombin.     

Association between dopamine D4 receptor (D4DR) exon III polymorphism and novelty seeking in Japanese subjects

Only five cases of paraganglioma of the prostate have been reported hitherto. The tumours may be functional (phaeochromocytomas) or non-functional and benign or malignant. We present a sixth case--a non-functional, benign paraganglioma of the prostate--with a review of the literature.