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( Suzuki Hirokazu ),( Shunji Takahashi ),( Hiroyuki Osada ),( Ken Ichi Yoshida ) 한국미생물 · 생명공학회 2011 Journal of microbiology and biotechnology Vol.21 No.7
DNA methylation in Streptomyces griseus IFO 13350 was analyzed by high-performance liquid chromatographic analysis and bisulfite-based analysis to reveal two methylation sites, 5`-GC5mCGGC-3` and 5`-GAG5mCTC-3`. The methylation was reconstituted in Escherichia coli by simultaneous expression of S. griseus SGR4675 and S. achromogenes M.SacI. The E. coli cells produced plasmids that mimicked the methylation profile of S. griseus DNA, which was readily introduced into S. griseus. The results of this study raise the possibility of a promising approach to establish efficient transformation in several streptomycetes.
( Suzuki Hirokazu ),( Ken Ichi Yoshida ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.9
We established an efficient transformation method for thermophile Geobacillus kaustophilus HTA426 using conjugative transfer from Escherichia coli of host-mimicking plasmids that imitate DNA methylation of strain HTA426 to circumvent its DNA restriction barriers. Two conjugative plasmids, pSTE33T and pUCG18T, capable of shuttling between E. coli and Geobacillus spp., were constructed. The plasmids were first introduced into E. coli BR408, which expressed one inherent DNA methylase gene (dam) and two heterologous methylase genes from strain HTA426 (GK1380-GK1381 and GK0343-GK0344). The plasmids were then directly transferred from E. coli cells to strain HTA426 by conjugative transfer using pUB307 or pRK2013 as a helper plasmid, pUCG18T was introduced very efficiently (transfer efficiency, 10-5-10-3 recipient-1), pSTE33T showed lower efficiency (10-7-10-6 recipient-1) but had a high copy number and high segregational stability. Methylase genes in the donor substantially affected the transfer efficiency, demonstrating that the host-mimicking strategy contributes to efficient transformation. The transformation method, along with the two distinguishing plasmids, increases the potential of G. kaustophilus HTA426 as a thermophilic host to be used in various applications and as a model for biological studies of this genus. Our results also demonstrate that conjugative transfer is a promising approach for introducing exogenous DNA into thermophiles.
TAKASHI SUZUKI,SHIGENOBU SHINOHARA,HIROFUMI YOSHIDA,HIROAKI IKEDA,YASUHIRO SAITOH,KEN-ICHI NISHIDE,MASAO SUMI 대한전자공학회 1992 대한전자공학회 학술대회 Vol.1992 No.10
An infrared range finder using a self-mixing laser diode (SM-LD), which has been proposed and developed by the authors, can measure not only a range of a moving target but its velocity simultaneously. In this paper, described is that the precise mode-hop pulse train can be obtained by employing a new signal processing circuit even when the backscattered light returning into the SM-LD is much more weaker. As a result, the distance to a tilted square sheet made from aluminium or white paper, which is placed 10 ㎝ through 60 ㎝ from the SM-LD, is measured with accuracy of a few percent even when the tilting angle is less than 75 degrees or 85 degrees, respectively.<br/> And in this paper, described is the range-image recognition of a plane object under the condition of standstill. The output laser beam is scanned by scanning two plane mirrors equipped with each stepping motor. And we succeeded in the acquisition of the range-image of a plane object in a few tens of seconds. Furthermore, described is a feasibility study about the range-image recognition of a slowly moving plane object.
Mahmood A. Hamed,Seiichi Nakata,Kazuya Shiogama,Kenji Suzuki,Ramadan H. Sayed,Yoichi Nishimura,Noboru Iwata,Kouhei Sakurai,Badawy S. Badawy,Ken-ichi Inada,Hayato Tsuge,Yutaka Tsutsumi 대한이비인후과학회 2017 Clinical and Experimental Otorhinolaryngology Vol.10 No.3
Objectives. Cholesteatoma is a nonneoplastic destructive lesion of the temporal bone with debated pathogenesis and bone resorptive mechanism. Both molecular and cellular events chiefly master its activity. Continued research is necessary to clarify factors related to its aggressiveness. We aimed to investigate the expression of Ki-67, cytokeratin 13 (CK13) and cytokeratin 17 (CK17) in acquired nonrecurrent human cholesteatoma and correlate them with its bone destructive capacity. Methods. A prospective quantitative immunohistochemical study was carried out using fresh acquired cholesteatoma tissues (n=19), collected during cholesteatoma surgery. Deep meatal skin tissues from the same patients were used as control (n=8). Cholesteatoma patients were divided into 2 groups and compared (invasive and noninvasive) according to a grading score for bone resorption based upon clinical, radiologic and intraoperative findings. To our knowledge, the role of CK17 in cholesteatoma aggressiveness was first investigated in this paper. Results. Both Ki-67 and CK17 were significantly overexpressed in cholesteatoma than control tissues (P<0.001 for both Ki-67 and CK17). In addition, Ki-67 and CK17 were significantly higher in the invasive group than noninvasive group of cholesteatoma (P=0.029, P=0.033, respectively). Furthermore, Ki-67 and CK17 showed a moderate positive correlation with bone erosion scores (r=0.547, P=0.015 and r=0.588, P=0.008, respectively). In terms of CK13, no significant difference was found between cholesteatoma and skin (P=0.766). Conclusion. Both Ki-67 and CK17 were overexpressed in cholesteatoma tissue and positively correlated with bone resorption activity. The concept that Ki-67 can be a predictor for aggressiveness of cholesteatoma was supported. In addition, this is the first study demonstrating CK17 as a favoring marker in the aggressiveness of acquired cholesteatoma.