- 著者
- Yoshizumi Ishino Takashi Ueno Masaru Miyagi Takashi Uemori Mitsuo Imamura Susumu Tsunasawa Ikunoshin Kato
- 出版者
- The Japanese Biochemical Society
- 雑誌
- The Journal of Biochemistry (ISSN:0021924X)
- 巻号頁・発行日
- vol.116, no.5, pp.1019-1024, 1994 (Released:2008-11-18)
- 参考文献数
- 18
We cloned the pol gene from the Thermus aquaticus YT-1 strain into a plasmid vector and constructed a high-level expression system of the gene in Escherichia coli. Six codons in the translational start region were changed to simple AT-type codons or codons which are most frequently used in E. coli by the genetic engineering techniques with retention of the amino acid sequence of the native enzyme. The modified pol genes were expressed under the lac promoter of pUC-type plasmid and 266, 418 units of activity was obtained in a sonicated and heated crude extract from 2g of E. coli cells bearing one of the recombinant plasmids, pTAQ9. Highly purified protein was subjected to structural analysis using a protein sequencer and an ion-spray mass spectrometer combined with reversed-phase HPLC (LC-MS). The primary structure of the DNA polymerase was identical with the amino acid sequence deduced from the nucleotide sequence of the pol gene as far as examined (about 95% of the sequence); though, several regions where small peptides of less than 5 residues were produced by lysyl endopeptidase digestion could not be sequenced.
- 著者
- Takashi Ueno Tokuya Omi Eiji Uchida Hiroyuki Yokota Seiji Kawana
- 出版者
- 日本医科大学医学会
- 雑誌
- Journal of Nippon Medical School (ISSN:13454676)
- 巻号頁・発行日
- vol.81, no.1, pp.4-11, 2014 (Released:2014-03-10)
- 参考文献数
- 23
- 被引用文献数
- 7 20
Background: Treating chronic wounds is challenging. Despite standard wound care, some chronic wounds fail to heal. Therefore, hyperbaric oxygen therapy (HBOT) was developed as an adjunct to standard wound care. Objective: To evaluate the efficacy of HBOT for treating chronic wounds due to a variety of causes at our institution. Methods: We reviewed the medical records of patients with chronic wounds treated with HBOT in addition to standard wound care at the Department of Dermatology, Nippon Medical School Hospital, from 2009 through 2012. Twenty-nine patients were reviewed (14 men and 15 women; mean age, 64.1±14.4 years). The cause of chronic wounds was diabetes mellitus (DM) in 13 patients, venous stasis in 10, polyarteritis nodosa cutanea in 2, and livedoid vasculopathy, pyoderma gangrenosum, chronic renal failure, and systemic sclerosis in 1 patient each. The patients underwent HBOT for 60 minutes with 100% oxygen delivered via a mask in a hyperbaric chamber pressurized to 2.8 atmospheres of absolute pressure. The response of the chronic wounds to HBOT was evaluated according to the following criteria: "excellent": more than 90% wound healing; "good": a greater than 30% reduction in wound size, and wound healing was confirmed on follow-up visits within 6 weeks; "fair": wound healing was achieved with a combination of further invasive interventions; and "poor": the wound showed a less than 30% reducion or worsened during HBOT, or wound healing had not been completed by follow-up visits within 6 weeks. Results: The response to HBOT was "excellent" in 6 patients, "good" in 8, "fair" in 11, and "poor" in 4. All 4 patients with a "poor" response had DM and had undergone hemodialysis. Conclusions: HBOT is an effective treatment for patients with chronic wounds, due to a variety of causes, when used in combination with conventional standard therapy or further interventions. However, HBOT is less effective in patients with DM than in patients with venous stasis because hemodialysis, which is more common in patients with DM, has negative effects on wound healing.