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Efficient Expression, Purification, and Characterization of a Novel FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus in Pichia pastoris

  • Yang, Yufeng;Huang, Lei;Wang, Jufang;Wang, Xiaoning;Xu, Zhinan
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1516-1524
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    • 2014
  • Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

Far Ultraviolet Observations of the Spica Nebula and the Interaction Zone

  • Choi, Yeon-Ju;Park, Jae-Woo;Lim, Tae-Ho;Min, Kyoung-Wook;Seon, Kwang-Il;Jo, Young-Soo
    • The Bulletin of The Korean Astronomical Society
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    • v.37 no.2
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    • pp.99.1-99.1
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    • 2012
  • We report the results of our analysis of far ultraviolet (FUV) observations made for the broad region around the ${\alpha}$ Vir (Spica) including the interaction zone of the Loop I and the Local Bubble. We employed the datasets of the GALEX and the FIMS, which made observations at similar FUV wavelengths. First, we noted that the GALEX image was enhanced in the southern region where the interaction zone exists. We attribute this enhanced FUV emission to dust scattering of the stellar photons, mostly from the background field stars with small contributions from the central star Spica. While the region is optically thin in general, the FUV intensity did not correlate well with the dust extinction level, indicating that the local radiation field has significant fluctuations. On the other hand, the GALEX FUV intensity well with the $H{\alpha}$ intensity as well as the dust extinction level in the northern part. In fact, the neutral hydrogen column density correlated very well with the dust extinction level throughout the whole region in consideration. The relationship between the neutral hydrogen column density and the color excess was estimated to be ${\sim}7{\times}10^{21}atoms\;cm^{-2}$, which is a little higher than the previous observations made for a diffuse interstellar medium. The spectral analyses of the FIMS observations showed the enhanced C IV emission throughout the whole region, indicating that the C IV emission arises by the interaction of the hot gases with the shell boundaries. A simple model showed that a large portion of the C IV emission comes from the Loop I side of the interaction zone, compared to the Local Bubble side. The FIMS spectrum also showed indications of the molecular hydrogen fluorescence lines for the interaction zone.

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Temperature Correction of Solar Radiation on Clear Sky Using by Modified Pyranometer (특수일사계를 이용한 맑은 날 일사량의 온도 보정)

  • Zo, Il-Sung;Jeong, Myeong-Jae;Lee, Kyu-Tae;Jee, Joon-Bum;Kim, Bu-Yo
    • Journal of the Korean Solar Energy Society
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    • v.35 no.1
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    • pp.9-19
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    • 2015
  • Pyranometer have many uncertainty factors (sensitivity function, thermal offset, other spectral effect, geometric, environment, and equipment etc.) than pyrheliometer. The solution for most of the uncertainty factors have been researched, but the problem for thermal offset is being continued research so far. Under the clear sky, due to the thermal offset of pyranometer, the diffuse and global radiation have been negative value for the nighttime and lower value for the daytime, respectively. In order to understand the uncertainty of the thermal offset effect, solar radiation are observed and analyzed using Ji and Tsay method and data from modified pyranometer. As a result of performing temperature correction using the modified pyranometer, the slope (dome factor; k) and intercept ($r_0$) from a linear regression method are 0.064 and $3.457g{\cdot}m^{-2}{\cdot}k^{-1}$, respectively. And the solar radiation is decreased significantly due to the effect of thermal offset during nighttime. The solar radiation from modified pyranometer increased approximately 8% higher than its observed by general pyranometer during daytime. By the way, these results did not generalize because its result is for only single case in clear sky. Accordingly, it is to required for accurate results obtained by the various cases (clear, cloudy and rainy) with longterm observations.

Leksell Frame-Based Stereotactic Biopsy for Infratentorial Tumor : Practical Tips and Considerations

  • Tae-Kyu Lee;Sa-Hoe Lim;Jangshik Jeong;Su Jee Park;Yeong Jin Kim;Kyung-Sub Moon;In-Young Kim;Shin Jung;Tae-Young Jung
    • Journal of Korean Neurosurgical Society
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    • v.67 no.2
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    • pp.249-256
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    • 2024
  • The Leksell frame-based transcerebellar approach was proposed with the arc support frame attached upside down to the Z coordinate. This study presented practical tips and considerations for obtaining adequate tissue samples for deep-seated cerebellar lesions or lower brainstem lesions specifically those accessible via the cerebellar peduncle. For practical insights, the Leksell coordinate frame G was fixed to prevent the anterior screw implantation within the temporalis muscle, to avoid interference with the magnetic resonance (MR)-adapter, and taking into account the magnetic field of MR in close proximity to the tentorium. After mounting of indicator box, the MR imaging evaluation should cover both the indicator box and the infratentorial region that deviated from it. The coordinates [X, Y, Za, Arc0, Ringa0] obtained from Leksell SurgiPlan® software (Elekta, Stockholm, Sweden) with arc 00 located on the patient's right side were converted to [X, Y, Zb=360-Za, Arc0, Ringb0=Ringa0-1800]. The operation was performed in the prone position under general anesthesia in four patients with deep cerebellar (n=3) and brainstem (n=1) tumors. The biopsy results showed two cases of diffuse large B-cell lymphoma, one metastatic braintumor and one glioblastoma. One patient required frame repositioning as a complication. Drawing upon the methodology outlined in existing literature, we anticipate that imparting supplementary expertise could render the stereotactic biopsy of infratentorial tumors more consistent and manageable for the practitioner, thereby facilitating adequate tissue samples and minimizing patient complications.

General Treatment Strategy for Intervention in Lower Extremity Arterial Disease (하지동맥 질환의 인터벤션: 전반적 치료 계획 수립)

  • Je Hwan Won
    • Journal of the Korean Society of Radiology
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    • v.82 no.3
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    • pp.500-511
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    • 2021
  • The prevalence of lower extremity disease is increasing with age. With recent technological advancements, endovascular treatment is being performed more frequently. The treatment goal of intermittent claudication is to improve walking and reduce claudication. To achieve these goals, anatomical durability and patency are important. In patients with critical limb ischemia, the lesions are diffuse and particularly severe in below-the-knee arteries. The treatment goal of critical limb ischemia is to promote wound healing and to prevent major amputation, which is evaluated by the limb salvage rate. Primary stenting using covered or bare metal stents is a widely accepted endovascular treatment. While drug-eluting technologies with or without atherectomy are widely used in the treatment of femoropopliteal disease, balloon angioplasty is the mainstay treatment for below-the-knee intervention. CT angiography provides a road map for planning endovascular treatment in patients without absolute contraindications.

Building Energy Savings due to Incorporated Daylight-Glazing Systems (통합 채광시스템의 건물 냉난방 에너지 성능평가)

  • Kim, Jeong-Tai;Ahn, Hyun-Tae;Kim, Gon
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.19 no.6
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    • pp.1-8
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    • 2005
  • The quantity of light available for a space can be translated in term of the amount of energy savings through a process of a building energy simulation. To get significant energy savings in general illumination, the electric lighting system must be incorporated with a daylight - activated dimmer control. A prototype configuration of an once interior has been established and the integration between the building envelope and lighting and HVAC systems is evaluated based on computer modeling of a lighting control facility. First of all, an energy-efficient luminaire system is designed and the lighting analysis program, Lumen-Micro 2000 predicts the optimal layout of a conventional fluorescent lighting future to meet the designed lighting level and calculates unit power density, which translates the demanded met of electric lighting energy. A dimming control system integrated with the contribution of daylighting has been applied to the operating of the artificial lighting. Annual cooling load due to lighting and the projecting saving amount of cooling load due to daylighting under overcast diffuse sky m evaluated by computer software ENER-Win. In brief, the results from building energy simulation with measured daylight illumination levels and the performance of lighting control system indicate that daylighting can save over 70 percent of the required energy for general illumination in the perimeter zones through the year A 25[%] of electric energy for cooling and almost all off heating energy may be saved by dimming and turning off the luminaires in the perimeter zones.

Expression of Oncogene Product in the Colorectal Carcinoma (결장암 및 직장암에서 암유전자 산물의 발현)

  • Shim, Young-Ran;Jang, Woo-Young;Choi, Kyoung-Chan;Choi, Joon-Hyuk;Choi, Won-Hee;Shim, Min-Chul
    • Journal of Yeungnam Medical Science
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    • v.12 no.2
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    • pp.210-225
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    • 1995
  • The expression of $p62^{c-myc}$ and $p21^{ras}$ can be seen in many solid tumor, but the pattern and incidence of expression were different according to organ, countries, and examiners, thus it is not definitely defined. Total 67 colorectal carcinoma in paraffin sections are analysed by immunohistochemically for evaluation of the $p62^{c-myc}$ and $p21^{ras}$ expression according to the age, sex, chief complaints, location, differentiation, modified Dukes stage, using the specific monoclonal antibodies. The results were summarized as follows : The age of patients ranged from 32 years to 82 years. The mean age was 57.6 years. The expression of $p62^{c-myc}$ and $p21^{ras}$ was not correlated with age. Male was 29 cases(43.3%) and female was 38 cases(56.7%). The male to female ratio was 1:1.31. The expression of $p21^{ras}$ was increased in female(p<0.05). Abdominal pain(43.7%) was the most frequent chief complaint. The most frequent tumor location was rectum(44.8%). The expression of $p62^{c-myc}$ was increased in the rectum(p<0.05). The 65 cases(97.0%) out of 67 cases showed positive reaction of $p62^{c-myc}$ in the nucleus, cytoplasmic membrane, and cytoplasm. The 62 cases(92.5%) out of 67 cases showed positive reaction of $p21^{ras}$ in the cytoplasmic membrane and cytoplasm. The positive rate of $p62^{c-myc}$ and $p21^{ras}$ was 97.0% and 91.4% in well differentiated adenocarcinoma, 100.0% and 95.0% in moderately differentiated adenocarcinoma, 90.0% and 90.0% in poorly differentiated adenocarcinoma, 100.0% and 100.0% in mucinous carcinoma. The positive rate of $p62^{c-myc}$ and $p21^{ras}$ was 94.1% and 88.2% in Dukes stage $B_1$, 96.0% and 96.0% in Dukes stage $B_2$, 100.0% and 100.0% in Dukes stage $C_1$, 100.0% and 88.9% in Dukes stage $C_2$, and 100.0% and 100.0% in Dukes stage D. The expression of $p62^{c-myc}$ in metastatic colorectal carcinoma showed diffuse and strongly positive reaction than primary colorectal carcinoma. The expression of $p21^{ras}$ in primary colorectal carcinoma showed diffuse and strongly positive reaction than metastatic colorectal carcinoma.

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A study on development of educational contents about combining computational thinking with design thinking (창의·컴퓨팅사고 교육내용 기본 설계 연구)

  • Oh, Kyung-Sun;Suh, Eung-Kyo;Chung, Haejin
    • Journal of Digital Convergence
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    • v.16 no.5
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    • pp.65-73
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    • 2018
  • The objective of this study is to suggest the contents of education for software for Undergraduates who are not majoring in IT to cultivate creative and convergent people to prepare for the 4th Industrial Revolution. In the background, this study investigated the relationship between design thinking and computing thinking factors and examined the direction of software education to develop creative problem solving abilities. Two specialist questionnaires and focus group interviews were used to derive two levels of content elements for design thinking based computing thinking. According to the results of the analysis, This research is based on the process of design thinking, focusing on diffuse thinking at the first level, and the whole contents are composed. The process of producing a prototype was used to derive the computational thinking contents necessary for problem solving. This study focuses on convergent thinking at the second level and draws out the contents of computing thought to embody the first level contents of various ideas and stories into computing. It is expected that this study will be used as basic data to develop the content and method of education as SW education for non - major subject in university.

A Case of Goodpasture's Syndrome Combined with Crohn's Disease (크론씨 병에 동반된 Goodpasture 증후군 1예)

  • Kim, Ji-Yon;Bae, Jun-Yong;Jung, Eun-Jung;Kim, Yang-Ki;Lee, Young Mok;Kim, Ki-Up;Uh, Soo-taek;Hwang, Jung-Hwa;Jin, So-Young;Lee, Dong-Wha
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.4
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    • pp.384-388
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    • 2006
  • A 29-year-old male patient was admitted due to his general weakness and poor oral intake for several months. He was diagnosed as having Crohn disease 16 years ago and total colectomy was performed 10 years ago. On the 3rd day after admission, gross hematuria and sudden hemoptysis combined with diffuse infiltration were noted on chest X-ray. His symptoms and the diffusely increased lung opacities improved with administering high-dose steroid therapy. Later, anti-GBM antibody was found to be positive on the laboratory findings. We report here on a rare case of Goodpsture syndrome combined with prolonged Crohn disease along with a review of literature.

A Clinical Analysis of Surgically Managed Thyroid Nodule (외과적으로 치료한 갑상선 결절에 대한 임상적 고찰)

  • Jung In-Kyu;Kim Lee-Su;Choi Won-Jin
    • Korean Journal of Head & Neck Oncology
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    • v.9 no.1
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    • pp.16-24
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    • 1993
  • During 7 years, from Jan. 1986 to Dec. 1992, authors studied 208 cases of the surgically managed thyroid nodules at the Department of General Surgery, Han Kang Sacred Heart Hospital and obtained the following results. 1) Among the total 208 cases, male to female ratio was 1:11.2 in benign thyroid diseases and 1:9 in malignant thyroid diseases. The benign disease was prevalent between second and forth decade comparing with malignant disease between third and fifth decade. 2) The most common duration of illness was 3 months(26.0%). 3) Palpable neck mass was the most common chief complaint(100%). Palpitation. fatigue, and sweating were common complaints in patient with benign disease, and fatigue, palpitation, and sweating in malignant disease in decreasing order of frequency. 4) 55.8 % of lesions were in right lobe, 33.2% in left lobe, 5.8% in diffuse type, 4.8% in bilateral lobes, and 0.5% in isthmus. 5) The most common size of nodule was between 2.0cm and 3.9cm in diameter, which consisted of 55.1% of benign disease and 48.0% of malignant disease. 6) 86.5% of thyroid function test showed euthyroidism, 10.1% hyperthyroidism, and 3.4% hypothyroidism. 7) Thyroid scanning of 176 patients revealed cold nodules in 92.5% of benign diseases and in 92.9% of malignant diseases. 8) The most common benign disease was adenomatous hyperplasia(62.7%), and the most common malignant disease was papillary adenocarcinoma(80.0%). 9) Fine needle aspiration cytology was performed in 91 cases, and it showed 69.0% of sensitivity, 90.3% of specificity, and 83.5% of accuracy. 10) Frozen biopsy was performed in 109 cases. and it showed 93.9% of sensitivity, 100.0% of specificity, and 98.2% of accuracy. 11) The most commonly performed operation was unilateral lobectomy(including unilateral lobectomy with isthmectomy)(79.1%) in benign disease. and total thyroidectomy(62.0%) in malignant disease. 12) Postoperative complication showed 5 cases of wound infection (2.4%), 3 cases of transient hypoparathyroidism(1.4%), 3 cases of transient hoarseness(1.4%), 2 cases of postoperative bleeding(1.0%), 1 case of permanent hypoparathyroidism(0.5%), 1 case of permanent hoarseness(0.5%), and 1 case of postoperative pneumonia (0.5%).

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