• Title/Summary/Keyword: Rosetta

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Cooperativity of ${\alpha}$- and ${\beta}$-Subunits of Group II Chaperonin from the Hyperthermophilic Archaeum Aeropyrum pernix K1

  • Kim, Jeong-Hwan;Lee, Jin-Woo;Shin, Eun-Jung;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.212-217
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    • 2011
  • ${\alpha}$ and ${\beta}$-subunits (ApCpnA and ApCpnB) are group II chaperonins from the hyperthermophilic archaeum Aeropyrum pernix K1, specialized in preventing the aggregation and inactivation of substrate proteins under conditions of transient heat stress. In the present study, the cooperativity of ${\alpha}$- and ${\beta}$-subunits from the A. pernix K1 was investigated. The ApCpnA and ApCpnB chaperonin genes were overexpressed in E. coli Rosetta and Codonplus (DE3), respectively. Each of the recombinant ${\alpha}$- and ${\beta}$-subunits was purified to 92% and 94% by using anionexchange chromatography. The cooperative activity between purified ${\alpha}$- and ${\beta}$-subunits was examined using citrate synthase (CS), alcohol dehydrogenase (ADH), and malate dehydrogenase (MDH) as substrate proteins. The addition of both ${\alpha}$- and ${\beta}$-subunits could effectively protect CS and ADH from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively, and MDH from thermal inactivation at $80^{\circ}C$C and $85^{\circ}C$. Moreover, in the presence of ATP, the protective effects of ${\alpha}$- and ${\beta}$-subunits on CS from thermal aggregation and inactivation, and ADH from thermal aggregation, were more enhanced, whereas cooperation between chaperonins and ATP in protection activity on ADH and MDH (at $85^{\circ}C$) from thermal inactivation was not observed. Specifically, the presence of both ${\alpha}$- and ${\beta}$- subunits could effectively protect MDH from thermal inactivation at $80^{\circ}C$ in an ATP-dependent manner.

Effect of Recombinant Olive Flounder Stanniocalcin on Serum Calcium Levels (혈청 칼슘 농도에 미치는 넙치 유전자 재조합 스타니오칼신의 효과)

  • Shin, Ji-Hye;Jung, Yu-Jung;Han, Yoon-Hee;Lee, Kyun-Young;Lee, Kyung-Mi;Kaneko, Toyoji;Sohn, Young-Chang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.4
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    • pp.307-313
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    • 2010
  • Stanniocalcin 1 (STC1) is a glycoprotein hormone that is important in the maintenance of calcium and phosphate homeostasis in both fish and mammals. STC1 and its paralog STC2 are expressed in multiple tissues in fishes, although the physiological roles of piscine STCs are still unclear compared with those of mammals. In this study, we cloned olive flounder STC1 (ofSTC1) and ofSTC2 cDNAs into pET28a vector and used E. coli Rosetta (DE3) as the host strain for protein expression. Expression experiments were carried out using isopropyl-$\beta$-D-thiogalactoside (IPTG) and nickel affinity chromatography. We could identify the recombinant proteins as single 29.5 kDa (ofSTC1) and 33.2 kDa (ofSTC2) bands in the insoluble fraction on sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). These results indicate that ofSTC1 and ofSTC2 were expressed as insoluble proteins in E. coli. Furthermore, the injection of ofSTC1 protein into juvenile tilapia resulted in a decrease of the serum calcium level. These results suggest that the purified fish STC1 and STC2 proteins may be used to elucidate the physiological role of STCs in fishes.

Expression and Characterization of Protein Latcripin-3, an Antioxidant and Antitumor Molecule from Lentinula edodes C91-3

  • Ann, Xiao-Hua;Lun, Yong-Zhi;Zhang, Wei;Liu, Ben;Li, Xing-Yun;Zhong, Min-Tao;Wang, Xiao-Li;Cao, Jing;Ning, An-Hong;Huang, Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.5055-5061
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    • 2014
  • In this study, an anti-oxidant and anti-tumor protein Latcripin-3 of Lentinula edodes C91-3 was expressed in Escherichia coli. for the first time. According to the cDNA library, the full-length gene of Latcripin-3 was cloned by the methods of 3'-full rapid amplification of cDNA Ends (RACE) and 5'-full RACE. The structural domain gene of Latcripin-3 was inserted into the pET32 a(+). The functional protein of Latcripin-3 was expressed in Rosetta-gami (DE3) E. coli, evaluated by Western blotting and mass spectrometry. DPPH testing showed that the protein Latcripin-3 can scavenge free radicals remarkably well. The activity of functional protein Latcripin-3 on A549 cells was studied with flow cytometry and the MTT method. The MTT assay results showed that there was a decreases in cell viability in a dose-dependent and time-dependent manner in protein Latcripin-3 treated groups. Flow cytometry demonstrated that Latcripin-3 can induce apoptosis and block S phase dramatically in human A549 lung cancer cells as compared to the control group. At the same time, the cell ultrastructure observed by transmission electron microscopy supported the results of flow cytometry. This research offers new insights and advantages for identifying anti-oxidant and anti-tumor proteins.

A Novel Approach to Cloning and Expression of Human Thymidylate Synthase

  • Lv, Ying-Tao;Du, Pei-Juan;Wang, Qiao-Yan;Tan, Yuan;Sun, Zong-Bin;Su, Zhong-Liang;Kang, Cong-Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7523-7527
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    • 2013
  • Thymidylate synthase (TS) catalyzes the transfer of a methyl group from methylenetetrahydrofolate to dUMP to form dTMP. It is a primary target in the chemotherapy of colorectal cancers and some other neoplasms. In order to obtain pure protein for analysis of structure and biological function, an expression vector TS-pET28b (+) was constructed by inserting wild-type human thymidylate synthase (hTS) cDNA into pET28b (+). Then an expression strain was selected after transformation of the recombined plasmid into Rosetta (DE3). Fusion protein with His-tag was efficiently expressed in the form of inclusion bodies after IPTG induction and the content was approximately 40.0% of total bacteria proteins after optimizing expression conditions. When inclusion bodies were washed, dissolved and purified by Ni-NTA under denatured conditions, the purity was up to 90%. On SDS-PAGE and West-blotting, the protein band was found to match well with the predicted relative molecular mass-36kDa. Bioactivity was 0.1 U/mg. The results indicated that high-level expression of wild-type hTS cDNA can be achieved in prokaryotes with our novel method, facilitating research into related chemotherapy.

Meaning of 'Labor' and Aesthetic in Dardenne Brothers' Films (다르덴 형제 영화에서 '노동'의 의미와 형식미학)

  • Ahn, Soong-Beum
    • The Journal of the Korea Contents Association
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    • v.12 no.9
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    • pp.93-105
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    • 2012
  • Dardenne brothers' films deals with minorities' issues who are shoved out to periphery of society, and especially takes interest in value-deprived human life in labor market of today. This study examines such social awareness of Dardenne brothers targeting their latest works awarded in Cannes Film Festival, , , , , . And analysis is attempted about Dardenne brothers' unique formal aesthetical style that exactly captures such social awareness. In Dardenne brothers' films a person shows up who faces abject matter of living. Their agony is closely connected to unemployment, and shows images of irresistible anxiety and alienation. Dardenne brothers' images are not only simple platform with contents embodied, but designed form showing essential thematic consciousness. Location and gaze, motion of camera capturing a character makes us to reflect the situation a character is facing in the world. There are almost no regard about plots, no dramatic devices, superficial and even straightforward, but Dardenne brothers' films have such independent formal aesthetic, which makes able to give endless tension.

Industrial Transformation in digital economy: A Case Study on PC and Comsumer Industries (디지털경제와 산업 전환: PC와 가전 산업의 사례 연구)

  • 배영자
    • Proceedings of the Technology Innovation Conference
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    • 2002.02a
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    • pp.133-149
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    • 2002
  • This study aims to investigate the impact of wide use of digital technology, in particular, the Internet, on innovation process and corporate strategy in electronics industry. The introduction of digital technology has changed innovation process, business model and organizational structure of the electronics companies. With the introduction of digital technology, the entire value chain of electronics industry from procurement, sales, and marketing to R&D and manufacturing has been restructured. E-commerce has been a major agenda for e-business. Recently, collaboration among electronics companies through e-marketplace has emerged as an important issue. A web-based e-commerce standard, so called RosettaNet, has been developed for facilitating e-transactions of electronics firms. The development of digital technology has dramatically increased the processing speed and sophisticated the virtual reality technology. As simulation becomes easier and more effective, the uncertainty and risk involved in R&D has decreased significantly. Another positive impact is closer cooperation between R&D and manufacturing functions. Taking advantage of automated and flexible production technology, has a new type of firm, so called, EMS (Electronics Manufacturing Services) emerged, whose strategic focus is on manufacturing only. The EMS can be seen as a kind of innovative organization, that is, a modular organization for production function. Digital technology has made convergence of computer and communication possible at early years but right now the convergence has been accelerated in extensive areas of communication, broadcasting, information appliances, software, contents, and services. Firms' effort for an innovative product and service has been intensified and the competition for a new standard product and service has become severe in electronics industry. Business activities are always realized in a specific organizational context. Accordingly building up innovation-friendly organization has emerged as a critical concern. Due to the striking decrease of transaction cost, a network type of organization has proliferated, and a business function turns into a modular organization. As a whole, digital technology has pushed electronics firms into developing their own business model, which takes consideration of standardization of business platform and their core competency.

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The effect of repeat processing on the color stability of lithium disilicate glass-ceramic (리튬디실리케이트 글라스-세라믹의 반복 성형이 색 안정성에 미치는 영향)

  • Jeon, Byung-Wook
    • Journal of Technologic Dentistry
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    • v.43 no.4
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    • pp.175-180
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    • 2021
  • Purpose: This study investigates the effect of repeated use of lithium disilicate glass-ceramics on color stability. Methods: In this study, 2 types of lithium disilicate glass-ceramics were used and they were divided into 4 groups after 1st, 2nd, 3rd, and 4th heat pressing. The L*, a*, and b* values of all specimens were measured using the VITA Easyshade Advance 4.0. These values was used to determine the ∆E value and transparency. IBM SPSS Statistics ver. 24.0, a statistical significance level of 0.05 and one-way ANOVA were used for all data. Results: In all groups, the L* and b* values were the lowest in the specimens subjected to the 4th heat pressing and the corresponding a* value was the highest. All specimens showed ∆E values of ≤3.3, indicating color changes that were not visible with the naked eye, and the color difference increased with the number of repeated heat pressing. The transparency of all specimens decreased as the number of repeated uses increased. Conclusion: According to the number of repeated heat pressing, the color difference of Rosetta SP (HASSBIO) was larger than that of IPS e.max Press (Ivoclar Vivadent), but it was clinically acceptable in all groups. Moreover, transparency decreased as the number of heat presses increased in all groups. According to the above findings, the lithium disilicate glass-ceramic has high color stability due to repeated heat pressing, implying that it can be used in clinical settings regularly.

Translucency and masking ability of translucent zirconia; comparison with conventional zirconia and lithium disilicate

  • Park, Joon Hee;Bang, Hyun Ji;Choi, Nak-Hyun;Park, Eun-Jin
    • The Journal of Advanced Prosthodontics
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    • v.14 no.5
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    • pp.324-333
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    • 2022
  • PURPOSE. The purpose of this study is to evaluate translucency and masking ability of translucent zirconia compared to conventional zirconia and lithium disilicate materials. MATERIALS AND METHODS. Three types of zirconia blocks with different yttria contents (3Y, 4Y, 5.5Y) and LS blocks (Rosetta SM) were used. Ten specimens for each group were fabricated with 10 mm diameter, with both 0.8 mm and 1.5 mm thicknesses (± 0.02 mm). All groups of zirconia specimens were sintered and polished according to the manufacturer's instructions. To calculate the translucency parameter (TP), CIELAB value was measured with a spectrophotometer on black and white backgrounds. To investigate the color masking abilities, background shades of A2, normal dentin, discolored dentin, and titanium were used. The color difference (ΔE) was calculated with the CIELAB values of A2 shade background as a reference compared with the values in the various backgrounds. One-way ANOVA and Bonferroni tests were conducted (P < .05). RESULTS. The TP values of zirconia specimens increased as the yttria content increased. All materials used in the study were able to adequately mask normal dentin shade (ΔE < 5.5), but were incapable of masking severely discolored dentin (ΔE > 5.5). On the titanium background, all materials of 1.5 mm thickness were able to mask the background shade, but with a thickness of 0.8 mm, only 3Y-TZP and 4Y-PSZ were able to mask titanium background. CONCLUSION. All zirconia materials and lithium disilicate specimens used in this study were unable to adequately mask the shade of severely discolored dentin. It is recommended to use 3Y-TZP or 4Y-PSZ with a sufficient thickness of 0.8 mm or more to mask titanium.

A Study on the Establishment of the Korean Women Doctor's Training Course in the Modern Period (근대시기 한국의 여의사 양성과정 성립 연구)

  • SHIN Eun-jeong
    • The Journal of Korean Medical History
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    • v.36 no.1
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    • pp.113-127
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    • 2023
  • The Gyeongseong Women's Medical Training Center was created as the result of the efforts of our internal visionaries with meaningful foreign missionaries to cultivate female doctors, yet the systematic structure of the institution developed primarily out of Korean efforts. Koreans have tried hard to cultivate their descendants and the skills of the Korean people within this framework, challenging the oppression of the ruling class in a given environment, and the results have continued to this day. First, during the Early period (1890-1909), Korea began to establish women's education and the first female doctors were trained with the help of foreign missionaries. Second, during the Growth period (1910-1919), while it was difficult for women's education to be easily expressed during Japanese colonial era, the need for women's education was growing as part of the patriotic enlightenment movement, and female students who wanted to become doctors began to go abroad. In addition, during this period, the means to train female doctors in Korea was available, but this system was not recognized by the Japanese colonial government. Third, during the Preparatory period (1920-1928), the Gyeongseong Women's Medical Class, which gave practical training to female doctors, was established and centered on Rosetta Hall and female doctors who studied abroad. Fourth, a women's medical school was established during the Establishment period (1929-1938), which created a foundation for stable supply of professional women's medical personnel. In this article, we studied the process of women who were marginalized in education until they were trained as professional intellectuals, and we hope that it will help them understand the current women's education in Korea and draw directions in the future.

Effect of machining precision of single ceramic restorations on the marginal and internal fit (단일 도재 수복물의 가공 정밀도가 변연 및 내면 적합도에 미치는 영향)

  • Son, Keunbada;Yu, Beom-Young;Lee, Kyu-Bok
    • The Journal of Korean Academy of Prosthodontics
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    • v.58 no.4
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    • pp.313-320
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    • 2020
  • Purpose: The purpose of this study was to evaluate the machining precision and the marginal and internal fit of single restorations fabricated with three types of lithium disilicate ceramic blocks and to evaluate the correlation. Materials and methods: Single restorations were designed using a CAD software program. The crown designed model file was extracted from the CAD software program. Three types of lithium disilicate blocks (Rosetta; HASS, IPS e.max CAD; Ivoclar vivadent, VITA Suprinity; VITA) were milled using a milling machine. For the fabrication of the crown scanned model file, the intaglio surface of the restoration was digitized using a contact scanner. Then, using the three-dimensional inspection software (Geomagic control X; 3D Systems), the process of the overlap of the crown designed model and the scanned model and 3-dimensional analysis was conducted. In addition, the marginal and internal fit of the crowns was evaluated by a silicone replication method. The difference among three types of single ceramic crown was analyzed using a Kruskal-Wallis H test, and Spearman correlation analysis was performed to analyze the correlation between machining precision and fitness (α=.05). Results: There was a significant difference in the machining precision and the marginal and internal fit according to the type of ceramic block (P<.001). In addition, the machining precision and the marginal and internal fit were positively correlated (P<.001). Conclusion: The marginal fit of crowns fabricated according to the types of ceramic blocks was within the clinically acceptable range (< 120 ㎛), so it can be regarded as appropriate machining precision applicable to all clinical as aspects in terms of the marginal fit.