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Protein Engineering of Flavin-containing Monooxygenase from Corynebacterium glutamicum for Improved Production of Indigo and Indirubin (인디고와 인디루빈의 생산을 증대하기 위한 플라빈-함유 모노옥시게나제의 단백질공학)

  • Jung, Hye Sook;Jung, Hae Bin;Kim, Hee Sook;Kim, Chang Gyeom;Lee, Jin Ho
    • Journal of Life Science
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    • v.28 no.6
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    • pp.656-662
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    • 2018
  • Flavin-containing monooxygenases from Corynebacterium (cFMOs) were mutagenized based on homology modeling to develop variants with an enhanced indigoid production capability. The four mutants, F170Y, A210G, A210S, and T326S, which fused to a maltose-binding protein (MBP), were constructed, and their biochemical properties were characterized. Of these, purified MBP-T326S required a higher concentration of exogenous FAD (100 mM) than the wild-type MBP-cFMO for optimal activity and showed a 3.8-fold increase in the $k_{cat}/K_m$ value at $100{\mu}M$ FAD compared to that of MBP-cFMO at $2{\mu}M$ FAD. The indole oxygenase activities of MBP-T326S decreased to 63-77% compared to that of the MBP-cFMO In addition, MBP-T326S displayed a very low level of futile NADPH oxidase activities (21-24%) in the absence of a substrate. Mutant proteins except for T326S displayed similar $K_m$ and increased $k_{cat}/K_m$ values compared to the wild-type. MBP-F170Y and -A210S mutants showed elevated indole oxygenase activity higher than 3.1- and 2.9-fold, respectively, in comparison with MBP-cFMO. When indigoid production was carried out in LB broth with 2.5 g/l of tryptophan, Escherichia coli expressing cFMO produced 684 mg/l of indigo and 104 mg/l of indirubin, while cells harboring T326S produced 1,040 mg/l of indigo and 112 mg/l of indirubin. The results indicate that the production of indigo was 13% higher when compared to a previous report in which an E. coli expressing FMO from Methylophaga produced 920 mg/l of indigo. The protein engineering of cFMO based on homology modeling provided a more rational strategy for developing indigoid-producing strains.

Evaluation of the Effect of Metal Artifacts Varying the Parameters of the Attenuation Map for the artificial Hip Joint in SPECT/CT (SPECT/CT에서 인공고관절에 대한 감쇠보정지도(Attenuation Map)의 매개변수 변화에 따른 금속 인공물(Metal Artifact)의 영향 평가)

  • Kim, Sang Gyu;Kim, Jung Yul;Park, Min Soo;Jo, Seung Hyun;Lim, Han Sang;Kim, Jae Sam
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.2
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    • pp.3-7
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    • 2014
  • Purpose SPECT/CT scan to be performed attenuation correction on the basis of CT induce an overestimation of the site due to the beam hardening artifact by metal cover and reduce the images quality. Therefore, this study using a phantom that has been inserted artificial hip joint investigated that effect on the SPECT/CT image causing by metal artifact for varying the parameters of the Attenuation Map. Materials and Methods Siemens Symbia T16 SPECT/CT equipment was used. Artificial hip joint was inserted to SPECT/PET phantom, 17 mm sphere of Bright Streak area in CT image was filled with Tc-99m so that the radiation activity was 8 times compared to background. And then Hot and Background was measured in varying Wide Beam Coefficient on Attenuation Map and RBR (Region to Background Ratio) of Metal and Non-Metal was calculated and analyzed depending on the presence or absence of the hip joint. Results It tended to hot count of Non-Metal and Metal to increase as the value of the manual mode is increased, hot count ratio with the group of both manual mode 0.5 and 0.4 is the best match. Also, in automatic mode, the ratio of RBRNon-Metal and RBRMetal was 1.135, statistically significant difference was not observed in the manual mode 0.5 and 0.4. Conclusion In the automatic mode of Wide Beam Coefficient in attenuation correction map, it was found that it is over-correction by 13.52%, it was possible to minimize the over-correction by the artifact in 0.5 and 0.4 of manual mode. Further studies should be performed in order to apply to a patient with the help of this and it is considered possible to reduce the over-correction by the metal artifact of an artificial hip joint for Hip-Resurfacing Arthroplasty patients, and to improve the diagnostic performance.

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Legal Interest in Damages Regarding Loss of Treatment Chance (치료기회상실로 인한 손해배상에 있어서 피침해법익)

  • Eom, Bokhyun
    • The Korean Society of Law and Medicine
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    • v.20 no.3
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    • pp.83-139
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    • 2019
  • Recognition of liability for damages due to medical malpractice has been developed largely on the basis of two paths. First is the case where there is an error in a physician's medical practice and this infringes upon the legal interests of life and body, and the compensation for monetary and non-monetary damages incurred from such infringement on life and body becomes an issue. Second is the case where there is a breach of a physician's duty of explanation that results in a infringement on the patient's right of autonomous decision, and the compensation for non-monetary damages incurred from such infringement becomes an issue. However, even if there is a medical error, since it is difficult to prove the causation between the medical error of a physician and the infringement upon legal interests, the physician's responsibility for damage compensation is denied in some cases. Consider, for example, a case where a patient is already in the final stage of cancer and has a very low possibility of a complete recovery even if proper treatment is received from the physician. Here, it is not appropriate to refuse recognition of any damage compensation based on the reason that the possibility of the patient dying is very high even in the absence of a medical error. This is so because, at minimum, non-monetary damage such as psychological suffering is incurred due to the physician's medical error. In such a case, our courts recognize on an exceptional basis consolation money compensation for losing the chance to receive proper treatment. However, since the theoretical system has not been established in minutiae, what comes under the benefit and protection of the law is not clearly explicated. The recent discourse on compensating for damages incurred by patients, even when the causation between the physician's medical error and infringement upon the legal interests of life and body is denied, by establishing a new legal interest is based on the "legal principle of loss of opportunity for treatment." On what should be the substance of the new legal interest, treatment possibility argument, expectation infringement argument, considerable degree of survival possibility infringement argument and loss of opportunity for treatment argument are being put forth. It is reasonable to see the substance of this protected legal interest as "the benefit of receiving treatment appropriate to the medical standard" according to the loss of opportunity for treatment argument. The above benefit to the patient is a value inherent to human dignity that should not be infringed upon or obstructed by anyone, and at the same time, it is a basic desire regarding life and a benefit worthy of protection by law. In this regard, "the benefit of receiving treatment appropriate to the medical standard" can be made concrete as one of the general personal rights related to psychological legal interest.

Etiology and Clinical Manifestations of Fever in Infants Younger than 3 Months Old: A Single Institution Study, 2008-2010 (3개월 미만 영아의 발열에 대한 원인 및 임상양상에 대한 단일기관 연구; 2008-2010)

  • Seok, Joon Young;Kang, Ji Eun;Cho, Eun Young;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.19 no.3
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    • pp.121-130
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    • 2012
  • Purpose : The purpose of this study is to investigate clinical features and causative organisms in febrile infants younger than three months, to help identification of high risk patients for serious bacterial infection (SBI). Methods : A total of 313 febrile infants younger than three months, who had visited Seoul National University Children's Hospital from January 2008 to December 2010 were included. Clinical features, laboratory findings, causative organisms, and risk factors of SBI were analyzed by retrospective chart review. Causative bacterial or viral pathogens were identified by gram stain and cultures, rapid antigen tests, or the polymerase chain reaction from clinically reliable sources. Results : Among 313 infants, etiologic organisms were identified in 127 cases (40.6%). Among 39 cases of bacterial infections, Escherichia coli (66.7%) and Streptococcus agalactiae (12.8%) were common. Enterovirus (33.7%), respiratory syncytial virus (19.8%), and rhinovirus (18.8%) were frequently detected in 88 cases of viral infection. Patients with SBI (39 cases) showed significantly higher values of the white blood cell count ($14,473{\pm}6,824/mm^3$ vs. $11,254{\pm}5,775/mm^3$, P=0.002) and the C-reactive protein ($6.32{\pm}8.51mg/L$ vs. $1.28{\pm}2.35mg/L$, P<0.001) than those without SBI (274 cases). The clinical risk factors for SBI were the male (OR 3.7, 95% CI 1.5-8.9), the presence of neurologic symptoms (OR 4.8, 95% CI 1.4-16.8), and the absence of family members with respiratory symptoms (OR 3.6, 95% CI 1.2-11.3). Conclusion : This study identified common pathogens and risk factors for SBI in febrile infants younger than three months. These findings may be useful to guide management of febrile young infants.

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A Study on the Effects of BCG Vaccination against Tuberculosis (BCG의 결핵예방(結核豫防) 효과에 관(關)한 연구(硏究))

  • Kang, Pock-Soo;Lee, Sung-Kwan
    • Journal of Preventive Medicine and Public Health
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    • v.15 no.1
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    • pp.33-46
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    • 1982
  • In Korea BCG vaccination has been employed as the main control measure for tuberculosis since 1962. Recently, the protective efficacy of BCG against tuberculosis has been controversial worldwide. A case-control study was conducted to evaluate the efficacy of BCG in Korea which has a high prevalence of tuberculosis(2.5%). The study subjects were children under the age of 14 years who were hospitalized and diagnosed as tuberculosis in three general hospitals in Taegu City during last 6 years ($1975{\sim}1980$). Among 416 hospitalized tuberculous patients, 314 cases were confirmed as to the presence or absence of the BCG scar. A control group was selected from the same hospital patients of the same period as the cases. The control group was other than tuberculous patients whose distribution of age, sex and residence were the same as the cases. The results obtained are as follows: For all forms of tuberculosis, the relative risk and the protective efficacy of BCG were 3.5 and 71.8%. The efficacy was higher among female than among male (78.6% vs. 65.8%). The efficacy was higher among the one year and above than among less than 1 year of age. For tuberculous meningitis, the relative risk and the protective efficacy of BCG were 3.9 and 74.3 %. The efficacy was statistically significantly higher among female than among male (p<0.05). The relative risk and the protective efficacyt of BCG for uberculous meningitis combined with miliary tuberculosis and combined with pulmonary tuberculosis represented 6.9, 85.6%, and 7.4, 86.5%, respectively. On the other hand, the relative risk and the protective efficacy of BCG for miliary tuberculosis were 2.1 and 51.6%, and for pulmonary tuberculosis, 2.3 and 54.7%, respectively. From these results, it appears that BCG vaccination is an efficient preventive measure in Korea where tubercluosis is prevalent. Thus the routine BCG vaccination should be continued.

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Prenatal Care Utilization Pattern and Its Determinants in Rural Korea (농촌지역 모성의 산전관리서비스 이용양상과 그 결정요인)

  • Kim, Jang-Rak;Park, Jung-Han;Lee, Jae-Kyong;Seo, Sang-Hong;Bang, Joon-Yong
    • Journal of Preventive Medicine and Public Health
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    • v.26 no.4 s.44
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    • pp.599-613
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    • 1993
  • To study the pattern of prenatal care utilization and its determinants in rural Korea, 976 mothers (65.5%) out of 1,489 living mothers in Chinyang, Sachon and Hapchon Counties in Kyongsangnam Province who had delivered a baby between July 1, 1990 and June 30, 1991 were interviewed by the Myon health workers from January 3 through February 15, 1992. The Anderson's behavioral model for health service utilization was applied to develop the frames for analysis. The dependent variable was a number of prenatal care visits. And the independent variables included In the model were the variables pertaining to the predisposing, enabling, medical need and other components. The proportion of mother who had ever received the prenatal care service for the index pregnancy was 97.3%. However, the proportion of mothers who had made more than 10 visits was only 20.6%, which indicated that majority of mothers had paid far less visits than recommended $10{\sim}12$ visits for each normal pregnancy. The low utilization of prenatal care services (none or less than 4 visits) was related to mother's low educational level, the high birth order, beneficiary of the medical aid, the absence of clinic in the community, no diagnosed disease of mother during pregnancy, and mothers engaged in farming. Inequity of access seemed to exist because social structure variables and the variables of enabling component were important predictors. And there seemed to be high mutability in equalizing the distribution of prenatal care services because the variables of enabling component such as type of medical security and whether there was a clinic or not in the community were substantially important.

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The Usability Evaluation of the Usefulness of Bismuth Shields in PET/CT Examination (PET/CT 검사에서 비스무스(bismuth) 차폐체의 적용에 따른 유용성 평가)

  • Park, Hoon-Hee;Lee, Juyoung;Kim, Ji-Hyeon;Nam, Kun-Sik;Lyu, Kwang-Yeul;Lee, Tae Soo
    • Journal of radiological science and technology
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    • v.37 no.1
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    • pp.49-56
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    • 2014
  • Recently with CT developed, various studies for reduction of exposure dose is underway. Study of bismuth shields in these studies is actively underway, and has already been applied in the clinical. However, the application of the PET/CT examination was not activated. Therefore, through this study, depending on the application of bismuth shields in the PET/CT examination, we identify the quality of the image and the impact on the Standard Uptake Value (SUV). In this study, to apply to the shielding of the breast, by using the bismuth shields that contains 0.06 mm Pb ingredients, was applied to the PET/CT GEMINI TF 64 (Philips Healthcare, Cleveland, USA). Phantom experiments using the NEMA IEC Body Phantom, images were acquired according to the presence or absence of bismuth shields apply. Also, When applying, images were obtained by varying the spacing 0, 1, 2 cm each image set to the interest range in the depth of the phantom by using EBW-NM ver.1.0. When image of the PET Emission acquires, the SUV was in increased depending on the use of bismuth shields, difference in the depth to the surface from deep in the phantom increasingly SUV increased (P<0.005). Also, when using shields, as the more gab decreased, SUV is more increased (P<0.005). Through this study, PET/CT examination by using of bismuth shields which is used as purpose of reduction dose. When using shields, the difference of SUV resulting from the application of bismuth shields exist and that difference when gab is decrease and surface is wider. Therefore, setting spacing of shield should be considered, if considering the reduction of the variation of SUV and image quality, disease of deep organs should be a priority rather than superficial organ disease. Use of bismuth shielding factor considering the standard clinical examination, decrease unnecessary exposure can be expected to be considered.

Kinematical Analysis of Heel-Brake Stop in Inline Skate (인라인 스케이트(Inline Skate) 힐 브레이크(Heel-Brake) 정지에 관한 운동학적 분석)

  • Han, Jae-Hee;Lim, Yong-Kyu
    • Korean Journal of Applied Biomechanics
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    • v.15 no.2
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    • pp.11-20
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    • 2005
  • This study has a purpose on contributing to apprehend safe and right way to stop to the inline skate beginners and to the instructors who teaches line skating on the basis for the result of the kinematical analysis on Heel brake stop movement of the inline skate, focusing on the displacement on COG, angle displacement of ankle joint, angle displacement of knee joint, angle displacement of hip joint, using a 3D image method by DLT. To achieve this goal, we analysed the kinematical factor of the 3 well-trained inline skating instructors and obtained the following results. 1. During the movement of heel-brake stop, when strong power was given to a stable and balanced stop and the lower limbs, if the physical centroid is lowered the stability increases, and if it is placed high from the base surface, as the stability decreases compared to the case of low physical centroid, we should make a stop by placing a physical centroid in the base surface and lowering the hight of physical centroid. 2. To make a stable and balanced stop and to provide a strong power to the lower limbs, it is advisable to make a stop by decreasing an angle displacement of ankle joint during a "down" movement. In case of the left ankle joint, in all events and phases the dorsiflexion angle showed a decrease. Nevertheless, in the case of the right ankle joint, the dorsiflexion angle shows an increase after a slight decrease. The dorsiflexion angle displacement of ankle joint can be diminished because of the brake pad of the rear axis frame of the right side inline skate by raising a toe, but cannot be more decreased if certain degree of an angle is made by a brake pad touching a ground surface. To provide a power to a brake pad, it is recommended to place a power by lowering a posture making the dorsiflexion angle of the left ankle joint relatively smaller than that of the right ankle. 3. To make a stable and balanced stop and to add a power to a brake pad, the power must be given to the lower limbs in lowering the hight of physical centroid. For this, it is recommended to make a down movement by decreasing the flexion angle of a knee joint and it is necessary to make a down movement by a regular decrease of the angle displacement of knee joint rather than a swift down movement in every event and phase. 4. The right angle displacement of hip joint is made by lowering vertically the hight of physical centroid as leaning slightly forward. If too narrow angle displacement of hip joint is made by leaning forward too much, the balance is lost during the stop by placing the center in front. To make a stable and balance stop and to place a strong power to the lower limbs, it is recommendable to make a narrow angle by lower the hip joint angle. However, excessive leaning of the upper body to make the angle too narrow, can cause an instable stop and loss of physical centroid. After this study, it is considered to assist the kinematical understanding during the heel brake stop movement of the inline skate, and, to present basic data in learning a method of stable and balanced stop for the inline skating beginners or for the inline skate instructors in the present situation of the complete absence of the study in inline skating.

Development and Evaluation of Traffic Conflict Criteria at an intersection (교차로 교통상충기준 개발 및 평가에 관한 연구)

  • 하태준;박형규;박제진;박찬모
    • Journal of Korean Society of Transportation
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    • v.20 no.2
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    • pp.105-115
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    • 2002
  • For many rears, traffic accident statistics are the most direct measure of safety for a signalized intersection. However it takes more than 2 or 3 yearn to collect certain accident data for adequate sample sizes. And the accident data itself is unreliable because of the difference between accident data recorded and accident that is actually occurred. Therefore, it is rather difficult to evaluate safety for a intersection by using accident data. For these reasons, traffic conflict technique(TCT) was developed as a buick and accurate counter-measure of safety for a intersection. However, the collected conflict data is not always reliable because there is absence of clear criteria for conflict. This study developed objective and accurate conflict criteria, which is shown below based on traffic engineering theory. Frist, the rear-end conflict is regarded, when the following vehicle takes evasive maneuver against the first vehicle within a certain distance, according to car-following theory. Second, lane-change conflict is regarded when the following vehicle takes evasive maneuver against first vehicle which is changing its lane within the minimum stopping distance of the following vehicle. Third, cross and opposing-left turn conflicts are regarded when the vehicle which receives green sign takes evasive maneuver against the vehicle which lost its right-of-way crossing a intersection. As a result of correlation analysis between conflict and accident, it is verified that the suggested conflict criteria in this study ave applicable. And it is proven that estimating safety evaluation for a intersection with conflict data is possible, according to the regression analysis preformed between accident and conflict, EPDO accident and conflict. Adopting the conflict criteria suggested in this study would be both quick and accurate method for diagnosing safety and operational deficiencies and for evaluation improvements at intersections. Further research is required to refine the suggested conflict criteria to extend its application. In addition, it is necessary to develope other types of conflict criteria, not included in this study, in later study.

Relationship between Incidence of Endometritis and Metabolic Status during Peri- and Postpartum Periods in Dairy Cows (젖소의 자궁내막염 발생과 분만 전·후 대사 상태와의 상관관계)

  • Jeong, Jae-Kwan;Choi, In-Soo;Kang, Hyun-Gu;Jung, Young-Hun;Hur, Tai-Young;Kim, Ill-Hwa
    • Journal of Veterinary Clinics
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    • v.32 no.5
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    • pp.426-432
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    • 2015
  • This study compared blood metabolites during peri- and postpartum periods among cows with clinical or subclinical endometritis and cows without endometritis. Blood samples from 207 Holstein dairy cows were collected at 4 weeks prepartum, just after calving, and at 1, 2, 4, and 6 weeks postpartum to measure serum concentrations of calcium, magnesium, non-esterified fatty acids (NEFAs), total cholesterol, albumin, urea nitrogen, ${\beta}$-hydroxybutyrate (BHBA), aspartate aminotransferase (AST), ${\gamma}$-glutamyltransferase, glucose, and phosphorus. Clinical endometritis was diagnosed by the observation of vaginal discharge (> 50% pus) and subclinical endometritis was diagnosed by the evaluation of uterine cytology (> 18% neutrophils) at 4 weeks postpartum. Cows were divided into three groups based on the presence or absence of clinical or subclinical endometritis: the control group (n = 104), the clinical endometritis group (n = 66), and the subclinical endometritis group (n = 37). Calcium and magnesium concentrations were lower in the clinical endometritis group than in the control and subclinical endometritis groups throughout the study period (p < 0.05 to 0.0001), whereas the NEFAs concentration was higher in the clinical endometritis group than in the control group throughout the study period (p < 0.01). The total cholesterol concentration was lower in the clinical endometritis group than in the control and subclinical endometritis groups throughout the pre- and postpartum periods (p < 0.05 to 0.001). The albumin concentration was lower in the clinical endometritis group than in the control and subclinical endometritis groups during the postpartum period (p < 0.05 to 0.001). The urea nitrogen concentration was lower in the clinical endometritis group than in the control and subclinical endometritis groups at 4 and 6 weeks postpartum (p < 0.01). At 1 week postpartum, the BHBA concentration was higher in the clinical endometritis group than in the control group (p < 0.05), whereas the AST concentration was higher in the clinical endometritis and subclinical endometritis groups than in the control group (p < 0.05). In conclusion, lower serum concentrations of calcium, magnesium, total cholesterol, albumin, and urea nitrogen, but higher concentrations of NEFAs, BHBA, and AST during the postpartum period were associated with the incidence of clinical endometritis, indicating the importance of balanced nutrition during the transition period.