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Jihye Seok,Kyung-Jin Kim 한국구조생물학회 2020 Biodesign Vol.8 No.3
Glycine oxidase (GO) is an enzyme that catalyzes the oxidation reaction of the primary and secondary amine of glycine. In this study, we overexpressed GO from Bacillus cereus ATCC 14579 (BcGO) and purified the protein to homogeneity by Ni-NTA affinity and size-exclusion chromatography. The BcGO protein was crystallized using hanging-drop vapordiffusion method in the presence of 15% (v/v) Tacsimate pH 7.0, 0.1 M HEPES pH 6.5, 6% (w/v) PEG 3350 at 293 K. X-ray diffraction data were collected to a maximum resolution of 2.36 Å. The BcGO crystals belong to the space group C2221 with unit cell parameters a = 82.18 Å, b = 132.81 Å, c = 165.212 Å, α = 90 °, β = 90 °, γ = 90 °. With two molecules of BcGO per asymmetric unit, the crystal volume per unit of protein mass is 2.74 Å3 Da-1, which correspond to a solvent content is approximately 55.82%.
Jihye Seok,Kyung-Jin Kim 한국구조생물학회 2022 Biodesign Vol.10 No.1
Feruloyl-CoA hydratase/lyase (FerB) is an enzyme that catalyzes the final step of converting ferulic acid to biovanillin. In this study, we overexpressed FerB from Pseudomonas putida KT2440 (PpFerB) and purified the protein by Ni-NTA affinity and size-exclusion chromatography. The PpFerB protein was crystallized using hanging-drop vapor-diffusion method in the presence of 25% 1,2-propanediol, 0.1 M sodium phosphate dibasic/citric acid pH 4.2, 5% (w/v) PEG 3000, and 10% (v/v) glycerol at 293 K. X-ray diffraction data were collected to a maximum resolution of 1.65 Å. The PpFerB crystals belong to the space group R3 with unit cell parameters a = 87.7 Å, b = 87.7 Å, c = 198.0 Å, α = 90°, β = 90°, γ = 120°. With six molecules of PpFerB per asymmetric unit, the crystal volume per unit of protein mass is 2.37 Å3 Da–1, corresponding to a solvent content of approximately 48.08%.
Jihye Kim,Wonseok Kang,Dong Hyun Sinn,Geum-Youn Gwak,Yong-Han Paik,Moon Seok Choi,Joon Hyeok Lee,Kwang Cheol Koh,Seung Woon Paik 대한간학회 2020 Clinical and Molecular Hepatology(대한간학회지) Vol.26 No.4
Background/Aims: Although hepatocellular carcinoma (HCC) is notorious for its high recurrence rate, some patients do not experience recurrence for more than 5 years after resection or radiofrequency ablation for early-stage HCC. For those with five recurrence-free period, the risk of HCC recurrence within the next 5 years remains unknown. Methods: A total of 1,451 consecutive patients (median, 55 years old; males, 79.0%; hepatitis B virus-related, 79.3%) with good liver function (Child-Pugh class A) diagnosed with early-stage HCC by Barcelona Clinic Liver Cancer Staging and received radiofrequency ablation or resection as an initial treatment between 2005 and 2010 were analyzed. Results: During a median follow-up period of 8.1 years, 961 patients (66.2%) experienced HCC recurrence. The cumulative recurrence rates increased to 39.7%, 60.3%, and 71.0% at 2, 5, and 10 years, respectively, and did not reach a plateau. Five years after HCC diagnosis, 487 patients were alive without experiencing a recurrence. Among them, during a median of 3.9 additional years of follow-up (range, 0.1–9.0 years), 127 patients (26.1%) experienced recurrence. The next 5-year cumulative recurrence rate (5–10 years from initial diagnosis) was 27.0%. Male sex, higher fibrosis-4 scores, and alpha-fetoprotein levels at 5 years were associated with later HCC recurrence among patients who did not experience recurrence for more than 5 years. Conclusions: The HCC recurrence rate following 5 recurrence-free years after HCC treatment was high, indicating that HCC patients warrant continued HCC surveillance, even after 5 recurrence-free years.
( Jihye Kim ),( Wonseok Kang ),( Dong Hyun Sinn ),( Geum-youn Gwak ),( Yong-han Paik ),( Moon Seok Choi ),( Joon Hyeok Lee ),( Kwang Cheol Koh ),( Seung Woon Paik ) 대한내과학회 2020 The Korean Journal of Internal Medicine Vol.35 No.1
Background/Aims: We systematically evaluated the clinical characteristics, prevalence of cirrhosis, and mode of detection in virus-unrelated (non-B non-C, NBNC) hepatocellular carcinoma (HCC) patients in Korea. Methods: A total of 447 consecutive treatment-naïve NBNC-HCC adult patients who were registered at the Samsung Medical Center HCC registry in Korea from 2010 to 2013 were analyzed. NBNC was defined as negative hepatitis B surface antigen and negative anti-hepatitis C virus antibody. Presence of cirrhosis was determined based on histological, radiological, endoscopic, and serologic results. Mode of detection was classified as either under surveillance, incidental, or symptomatic. Results: Heavy alcohol use was the most common potential etiology in NBNC-HCC (NBNC-A, alcohol) (59.7%). Ten patients had other identif iable causes (NBNC-O, other identifiable cause) such as autoimmune hepatitis. The rest (38.0%) had no-identifiable cause (NBNC-NA-NO, non-alcohol, no-other identifiable cause). In NBNC-NA-NO group, 83.5% (96/115) of patients with available hepatitis B core immunoglobulin G antibody (HBcIgG) showed HBcIgG positivity, and 80.6% (137/170) had metabolic risk factors (diabetes, obesity, hypertension, and/ or dyslipidemia). Cirrhosis was present in 90.0%, 70.4%, and 60.0% of NBNC-O, NBNC-A, and NBNC-NA-NO patients, respectively. The proportion of patients diagnosed under surveillance was 25.5% across all patients, with specific proportions being 80.0%, 27.7%, and 18.8% for NBNC-O, NBNC-A, and NBNC-NA-NO, respectively. Conclusions: Among NBNC-HCC patients, heavy alcohol use or any other identifiable cause was not found in 38.0%. These NBNC-NA-NO HCC patients showed a high prevalence of HBcIgG positivity and metabolic risk factors, suggesting that prior hepatitis B virus infection and metabolic risk factors may be major contributing factors in the hepatocarcinogenesis in NBNC-NA-NO patients.
Effects of Repeated Ovarian Stimulation on Ovarian Function and Aging in Mice
Jihye Whang,Cheyoung Ahn,Soohyun Kim,Eunji Seok,Yunjeong Yang,Goeun Han,Haeun Jo,양현원 한국발생생물학회 2021 발생과 생식 Vol.25 No.4
Controlled ovarian hyperstimulation (COH) is routinely used in the in vitro fertilization and embryo transfer (IVF-ET) cycles to increase the number of retrieved mature oocytes. However, the relationship between repeated COH and ovarian function is still controversial. Therefore, we investigated whether repeated ovarian stimulation affects ovarian aging and function, including follicular development, autophagy, and apoptosis in follicles. Ovarian hyperstimulation in mice was induced by intraperitoneal injection with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Mice subjected to ovarian stimulation once were used as a control group and 10 times as an experimental group. Repeated injections with PMSG and hCG significantly reduced the number of primary follicles compared to a single injection. The number of secondary and antral follicles increased slightly, while the number of corpus luteum increased significantly with repeated injections. On the other hand, repeated injections did not affect apoptosis in follicles associated with follicular atresia. The expression of autophagy-related genes Atg5, Atg12, LC3B, and Beclin1, cell proliferation-related genes mTOR, apoptosis-related genes Fas, and FasL was not significantly different between the two groups. In addition, the expression of the agingrelated genes Dnmt1, Dnmt3a, and AMH were also not significantly different. In this study, we demonstrated that repeated ovarian stimulation in mice affects follicular development, but not autophagy, apoptosis, aging in ovary. These results suggest that repetition of COH in the IVF-ET cycle may not result in ovarian aging, such as a decrease in ovarian reserve in adult women.