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1.  Crystal structure of endo-1,4-β-glucanase from Eisenia fetida  
Journal of Synchrotron Radiation  2013;20(Pt 6):884-889.
The crystal structure of endo-1,4-β-glucanase from the earthworm Eisenia fetida, which retained sufficient cellulase activity at low temperature, was determined at 1.5 Å resolution.
The saccharification process is essential for bioethanol production from woody biomass including celluloses. Cold-adapted cellulase, which has sufficient activity at low temperature (<293 K), is capable of reducing heating costs during the saccharification process and is suitable for simultaneous saccharification and fermentation. Endo-1,4-β-glucanase from the earthworm Eisenia fetida (EF-EG2) belonging to glycoside hydrolase family 9 has been shown to have the highest activity at 313 K, and also retained a comparatively high activity at 283 K. The recombinant EF-EG2 was purified expressed in Pichia pastoris, and then grew needle-shaped crystals with dimensions of 0.02 × 0.02 × 1 mm. The crystals belonged to the space group P3221 with unit-cell parameters of a = b = 136 Å, c = 55.0 Å. The final model of EF-EG2, including 435 residues, two ions, seven crystallization reagents and 696 waters, was refined to a crystallographic R-factor of 14.7% (free R-factor of 16.8%) to 1.5 Å resolution. The overall structure of EF-EG2 has an (α/α)6 barrel fold which contains a putative active-site cleft and a negatively charged surface. This structural information helps us understand the catalytic and cold adaptation mechanisms of EF-EG2.
PMCID: PMC3795549  PMID: 24121333
endoglucanase; glycoside hydrolase family 9; cold-adapted enzyme; Eisenia fetida
2.  Auricular Acupuncture at the “Shenmen” and “Point Zero” Points Induced Parasympathetic Activation 
Purpose. Since auricular acupuncture is a diagnostic and treatment system based on normalizing the body's dysfunction, auricular acupuncture has been applied for pain relief, relaxation, and so on. These techniques would modulate the autonomic nerve system, thereby inducing the above-mentioned effects. The aim was to see the effect of auricular acupuncture applied to the “Shenmen” and “Point Zero” points on the postoperative heart rate variability (HRV). Methods. Twenty-six patients who underwent hemicolectomy under general anesthesia were randomized into the control or the acupuncture group. After the operation and before emergence, the acupuncture group received auricular acupuncture. An electrocardiographic unit was placed for recording the autonomic nervous activities. Results. The low frequency (LF)/high frequency (HF) ratio of HRV increased (P = 0.0007) in the control, but the ratio in the acupuncture did not change. There were significant differences between the ratios of the two groups at 3 : 00, 4 : 00, and 5 : 00. HF of the acupuncture group tended to be higher. HFs of the acupuncture group were significantly higher than those of the control group at 3 : 00, 4 : 00, and 5 : 00. Conclusion. Auricular acupuncture kept the LF/HF ratio at lower levels and HF at higher levels during postoperative period in the patients who had undergone hemicolectomy.
PMCID: PMC3687596  PMID: 23861718
3.  Comprehensive predictions of target proteins based on protein-chemical interaction using virtual screening and experimental verifications 
BMC Chemical Biology  2012;12:2.
Identification of the target proteins of bioactive compounds is critical for elucidating the mode of action; however, target identification has been difficult in general, mostly due to the low sensitivity of detection using affinity chromatography followed by CBB staining and MS/MS analysis.
We applied our protocol of predicting target proteins combining in silico screening and experimental verification for incednine, which inhibits the anti-apoptotic function of Bcl-xL by an unknown mechanism. One hundred eighty-two target protein candidates were computationally predicted to bind to incednine by the statistical prediction method, and the predictions were verified by in vitro binding of incednine to seven proteins, whose expression can be confirmed in our cell system.
As a result, 40% accuracy of the computational predictions was achieved successfully, and we newly found 3 incednine-binding proteins.
This study revealed that our proposed protocol of predicting target protein combining in silico screening and experimental verification is useful, and provides new insight into a strategy for identifying target proteins of small molecules.
PMCID: PMC3471015  PMID: 22480302
4.  Transcutaneous Electrical Nerve Stimulation on the PC-5 and PC-6 Points Alleviated Hypotension after Epidural Anaesthesia, Depending on the Stimulus Frequency 
Neuraxial blockade causes arterial hypotension. Transcutaneous electrical nerve stimulation (TENS) at the Neiguan (PC-6) and Jianshi (PC-5) reduces the severity of hypotension after spinal anaesthesia, but did not clarify the optimal stimulus frequency. We hypothesized that the stimulus frequency of TENS at the PC-6 and PC-5 points would influence the severity of hypotension after epidural anaesthesia. 65 ASA I or II male patients presenting for inguinal hernia repair were randomized to five groups: the control group received no treatment; the 2 Hz, 10 Hz, 20 Hz, and 40 Hz groups received TENS at a frequency of 2 Hz, 10 Hz, 20 Hz, and 40 Hz, respectively. The lowest SBP was significantly higher in the 40 Hz group [the control, 84 (74–110) mmHg; the 2 Hz, 96 (62–116) mmHg; the 10 Hz, 100 (68–110) mmHg; the 20 Hz, 96 (64–115) mmHg; the 40 Hz, 104 (75–140) mmHg: P = 0.004]. Significantly less patients experienced hypotension in the 40 Hz group [the control, 78%; the 2 Hz, 43%; the 10 Hz, 38%; the 20 Hz, 38%; the 40 Hz, 8%: P = 0.008]. TENS on the PC-6 and PC-5 points reduced the severity and incidence of hypotension after epidural anaesthesia, depending on the stimulus frequency.
PMCID: PMC3348533  PMID: 22611431
5.  Auricular Acupunctures are Effective for the Prevention of Postoperative Agitation in Old Patients 
Postoperative cognitive problems and delirium are not uncommon in the elderly. We reported four cases in which auricular acupunctures on the ‘Shenmen’ and ‘Point Zero’ points successfully managed postoperative problematic behaviors of the three patients with dementia and the one patient postoperatively demonstrating an agitated behavior.
PMCID: PMC2887336  PMID: 19875431
agitation; auricular acupuncture; delirium; dementia
6.  Enhanced HSP70 lysine methylation promotes proliferation of cancer cells through activation of Aurora kinase B 
Nature Communications  2012;3:1072-.
Although heat-shock protein 70 (HSP70), an evolutionarily highly conserved molecular chaperone, is known to be post-translationally modified in various ways such as phosphorylation, ubiquitination and glycosylation, physiological significance of lysine methylation has never been elucidated. Here we identify dimethylation of HSP70 at Lys-561 by SETD1A. Enhanced HSP70 methylation was detected in various types of human cancer by immunohistochemical analysis, although the methylation was barely detectable in corresponding non-neoplastic tissues. Interestingly, methylated HSP70 predominantly localizes to the nucleus of cancer cells, whereas most of the HSP70 protein locates to the cytoplasm. Nuclear HSP70 directly interacts with Aurora kinase B (AURKB) in a methylation-dependent manner and promotes AURKB activity in vitro and in vivo. We also find that methylated HSP70 has a growth-promoting effect in cancer cells. Our findings demonstrate a crucial role of HSP70 methylation in human carcinogenesis.
HSP70 is a molecular chaperone that aids protein folding. In this study, HSP70 is shown to be methylated and this post-translationally modified protein is elevated in expression in human cancers and promotes the activity of Aurora kinase B.
PMCID: PMC3658001  PMID: 22990868

Results 1-6 (6)