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Effect of Timing and Placement of N Fertilizer Application for Increased Use Efficiency - Principle and Practice (열대지역(熱帶地域)에 있어서 질소비료(窒素肥料)의 시용시기(施用時期)와 시비위치(施肥位置)가 비료효율(肥料效率)에 미치는 영향(影響) - 원리(原理)와 실제(實際))

  • Hong, Chong-Woon
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.3
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    • pp.285-299
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    • 1987
  • Timing and placement of fertilizer applications are two managerial means to improve the fertilizer use efficiency. The relative importance of these two means is determined by the application rate. With the realistic rate of N application recommended to the small farmers in the tropics, at present and in the near future, basal application in right manner, seems to be more important than split application at different times. In wetland rice soils, deep placement by whatever available means is desirable. But in the situations where perfect deep placement is very difficult to implement, the whole-layer application may be worth trying, until better methods become available. In rainfed uplands, N fertilizer application plans should be contingent upon the amount and distribution of rainfall: apply a less risky rate as subsurface banding near the crop rows to start with; then, depending upon the rainfall prospects in the season, apply or omit the additional dose. Because the patterns of crop response to N fertilizer can be significantly different between the research farms and farmers' fields, it seems imperative to have information on the patterns of crop response to N under farmers' management conditions, for the development of realistic fertilizer application recommendations. To enable the farmers to adopt improved fertilizer application technologies, it is essential to develop and make available to farmers convenient fertilizer applicators. Past experience with the improved fertilizer use technologies indicates that, in the long run, the development of fertilizers that are not only effective and convenient for farmers to use but also easy to produce without major modifications of existing fertilizer production systems is the ultimate solution to the problem of low N fertilizer use efficiency.

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A Literal Study of Feature of the Preventive Medicine in Oriental Medical Science (한의(韓醫) 예방의학(豫防醫學)의 특징(特徵)에 관(關)한 문헌적(文獻的) 고찰(考察))

  • Lee Sang-Woon;Lee Sun-Dong
    • Journal of Society of Preventive Korean Medicine
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    • v.1 no.1
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    • pp.85-104
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    • 1997
  • As the concern about health is increased, the importance of preventive medicine that prevent disease in advance to overcome boundary of disease remedy gets emphasized in the whole world. The fundamental thoughts of oriental medicine are the harmony between the human body and nature, the unified idea regarding mind in the same light with body, and the symmetry of the cosmo dual forces. And oriental medicine is a kind of study that has developed on the ground of prevention thought. from old days, it has been developed the preventive medicine at is called 'Yangseng(養生; recuperation)' etc. with the clinical medicine. The preventive medicine of oriental medicine was taking a serious of the first preventive medicine of an incurable disease thought and the prevention thought that belongs to the second and third preventive medicine is described through the whole oriental medicine. Also the clinical and preventive medicine to apply to the clinics have an in- separable relation. Therefore I inspected the part of the preventive medicine described in some oriental medicine books and studied the characteristics of preventive medicine of oriental medicine as follows; First, the preventive medicine of oriental medicine has the characteristics that is emphasized generally in the first preventive medicine and wholly it is composed in the system of the first, second and third preventive medicine. It has the presentative theory such as 'Jungkijonae sabulkaghan(正氣存內 邪不可干; If good energy is in body, a disease. dosen't occur)', 'Husajukpung Phijeyushi(虛邪賊風 避之有時; When e infectious disease like plague break out, they must avoid the place occurring the disease)', 'Chuninsangeung(天人相應; The harmony of nature and human)' etc.. This is intimately related to the incurable disease thought of the inside diameter and oriental medicine has pursued that. Second, due to the most prerequisite theory of disease production, the balanced condition of environment, the cause of a disease and host is called the health. As oriental medicine has the system of aetiology like that, we can see the host and environments are importance most of all. Namely we can think of the relation of host >> environment > the cause a disease Up to date Jungkijonae Sabulkaghan(正氣存內 邪不可干) that the most oriental medicine doctors have had a knowledge is not whole theory but a pan of oriental medicine science and it is included in oriental medicine theory to avoid infectious disease such as Husajukpung Phijeyushi(虛邪賊風 避之有時). Third, according to the natural result of the first and second contents, we can know that its characteristics stress the remedy without drugs. Because Jungkijonae Sabulkaghan(正氣存內 邪不可干), Husasukpung Phijeyushi(虛邪賊風 避之有時), and Chuninsangeung(天人相應) mean that they prevent disease in condition of freedom from ailment, We can prevent the disease production through the positive preventive methods such as exercise, spirit, food and innate prevention etc.. fourth, the preventive medicine of oriental medicine has developed with therapeutics and it contains all oriental medical methods such as host, environment, exercise, acupuncture and moxibustion, innate or postnatal methods. Also it is the general preventive medicine that has fundamental philosophy of oriental medicine; for examples, the unified idea, the harmony of nature and human, and the unity of mind and body. fifth, to develope the above scientific merit and special features, the preventive medicine of Oriental medicine must be objectified and reemergent stud? gets more and more essential from now on. Especially we need to have the scientific concern of Oriental medicine about the cause of a disease, environmental hygiene, industrial sanitation, and personal hygiene etc..

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Chemical Structures and Physiological Activities of Plant Growth Substance, Malformin B's (식물생장조절물질 말포민 B동족체의 화학구조 및 생리활성)

  • Kim, K.W.
    • Korean Journal of Weed Science
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    • v.15 no.1
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    • pp.85-98
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    • 1995
  • Six malformin B's produced by Aspergillus niger van Tiegh. were separated by HPLC. Their structures determined by the methods of amino acid analyses, mass spectrometry, and two-dimensional NMR were revealed as cyclic pentapeptides structurally related to malformin $A_1$. Both the NMR and MS/MS data suggest that the respective structures of separated malformin B's were as follows; cyclo-D-Cys-D-Cys-L-Val-D-Leu-L-allo-Ile for $B_{1a}$, cyclo-D-Cys-D-Cys-L-Val-D-Leu-L-Leu for $B_{1b}$, cyclo-D-Cys-D-Cys-L-Val-D-Val-L-Leu for $B_2$, cyclo-D-Cys-D-Cys-L-Val-D-Ile-L-Leu for $B_3$, cyclo-D-Cys-D-Cys-L-Val-D-Ile-L-Ile for $B_4$, and cyclo-D-Cys-D-Cys-L-Val-D-Val-L-Ile for $B_5$. Among the malformin B's, the structure of $B_{1b}$ was the same as that of malformin $A_3$ or C. All the malformin B's showed physiological activities in the two assay systems using corn(Zea mays L.) roots and mung bean(Phaseolus aureus Roxb.) hypercotyl segments. The malformin B's with molecular weight 529 were more effective for inducing corn root curvature than those with molecular weight 515. The difference in molecular weight of malformin B's, i.e., the retention time on HPLC, results in the polarity change of the whole malformin molecule which affects the revealation of the malformin activities. In addition, the disulfide form of the malformin B's gives the rigidity of the molecule, whereas the combination of the fourth and the fifth amino acid residues provides the optimal three-dimensional configuration to the malformin receptor of plants. Presumably, these two factors are appeared to be essential for the greatest physiological activity of malformin B's. malformin $B_{1a}$ caused the corn root curvature by 90% at a concentration of $0.25{\mu}M$. However, such differential activities with molecular weight of 529 or 515 of malformin B's were not found in the mung bean hypercotyl segment test. Maximum stimulation of mung bean hypercotyl growth was observed at $0.1{\mu}M$ concentration of malformin B's. The growth of the segments treated with $B_5$ was 154% greater than that of the control.

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Surgical Treatment for Carotid Artery Stenosis (경동맥 협착증의 수술적 치료)

  • Kim, Dae-Hyun;Yi, In-Ho;Youn, Hyo-Chul;Kim, Bum-Shik;Cho, Kyu-Seok;Kim, Soo-Cheol;Hwang, Eun-Gu;Park, Joo-Chul
    • Journal of Chest Surgery
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    • v.39 no.11 s.268
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    • pp.815-821
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    • 2006
  • Background: Carotid endarterectomy is an effective treatment modality in patients with severe carotid artery stenosis, but it may result in serious postoperative complications, We analyzed the results of the carotid endarterectomy performed in our institution to reduce the complications related to the card endarterectomy. Material and Method: We analyzed retrospectively the medical records of 74 patients(76 cases) who underwent carotid endarterectomy for carotid artery stenosis by a single surgeon from February 1996 to July 2004. Result: There were 64 men and 10 women. The mean age of the patients was 63.6 years old. Carotid endarterectomy only was performed in 63 cases, carotid endarterectomy with patch angioplasty in 8 cases, and carotid endarterectomy with segmental resection of internal carotid artery and end to end anastomosis in 5 cases. Intra-arterial shunt was used in 29 cases. The mean back pressures of internal carotid arteries checked after clamping common carotid arteries and external carotid arteries were $23.48{\pm}10.04$ mmHg in 25 cases with changes in electroencephalography(group A) and $47.16{\pm}16.04$ mmHg in 51 cases without changes in electroencephalography(group B). There was no statistical difference in the mean back pressure of internal carotid arteries between two groups(p=0.095), but the back pressures of internal carotid arteries of all patients with changes in electroencephalography were under 40 mmHg. When there was no ischemic change of electroencephalography after clamping common carotid artery and external carotid artery, we did not make use of intra-arterial shunt regardless of the back pressure of internal carotid artery. Operative complications were transient hypoglossal nerve palsy in four cases, cerebral hemorrhage occurred at previous cerebral infarction site in two cases, mild cerebral infarction in one case, hematoma due to anastomosis site bleeding in one case, and upper airway obstruction due to laryngeal edema probably caused by excessive retraction during operation in two cases. One patient expired due to cerebral hemorrhage occurring at previous cerebral infarction site. Conclusion: Carotid endarterectomy is a safe operative procedure showing low operative mortality. We suggest that intra-arterial shunt usage should be decided according to the ischemic change of electroercephalography regardless of the back pressure of internal carotid artery. Excessive retraction during operation should be avoided to prevent upper alway obstruction due to laryngeal edema and if upper airway obstruction is suspected, prompt management is essential.

Expression of Hypoxia-inducible Factor-$1{\alpha}$ in Non-small Cell Lung Cancer: Relationship to Prognosis and Tumor Biomarkers (비소세포 폐암에서 HIF-$1{\alpha}$의 발현: 예후 및 종양표지자와의 관련성)

  • Cho, Sung-Rae;Byun, Joung-Hun;Kim, Jong-In;Lee, Bong-Geun;Chun, Bong-Kwon
    • Journal of Chest Surgery
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    • v.39 no.11 s.268
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    • pp.828-837
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    • 2006
  • Background: Tissue hypoxia is characteristic of many human malignant neoplasm, and hypoxia inducible factor-1(HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increasing $O_2$ delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-$1{\alpha}$ has been reported in many human malignancies, but in non-small cell lung carcinoma the influence of HIF-$1{\alpha}$ on tumor biology, including neovascularization, is not still defined. In present study the relationship of HIF-$1{\alpha}$ expression on angiogenetic factors, relationship between the tumor proliferation and HIF-$1{\alpha}$ expression, interaction of HIF-$1{\alpha}$ expression and p53, and relationship between HIF-$1{\alpha}$ expression and clinico-pathological prognostic parameters were investigated. Material and Method: Archival tissue blocks recruited in this study were retrieved from fifty-nine patients with primary non-small cell lung carcinoma, who underwent pneumonectomy or lobectomy from 1997 to 1999. HIF-$1{\alpha}$, VEGF(vascular endothelial growth factor), and p53 protein expression and Ki-67 labeling index in tumor tissues were evaluated, using a standard avidin-biotin-peroxidase complex(ABC) immunohistochemistry. Relationship between the HIF-$1{\alpha}$ expression and VEGF, p53 overexpression and correlation between the HIF-$1{\alpha}$ expresseion and Ki-67 index were analyzed. Clinico-pathologic prognostic parameters were also analyzed. Result: HIF-$1{\alpha}$ expression in cancer cells was found in 24 of 59 cases of non-small cell lung carcinoma(40.7%). High HIF-$1{\alpha}$ expression was significantly associated with several pathological parameters, such as pathological TMN stage(p=0.004), pT stage(p=0.020), pN stage (p=0.029), and lymphovascular invasion(p=0.019). High HIF-$1{\alpha}$ expression was also significantly associated with VEGF immunoreactivity(p<0.001), and aberrant p53 expression(p=0.040). but was marginally associated with Ki-67 labeling index(p=0.092). The overall 5-year survival rate was 42.3%. The survival curve of patients with a high HIF-$1{\alpha}$ expression was worse than that of patients with low-expression(p=0.002). High HIF-$1{\alpha}$ expression was independent unfavorable factors with a marginal significance in multivariate analysis performed by Cox regression. Conclusion: It is suggested that high HIF-$1{\alpha}$ expression may be associated with intratumoral neovascularization possibly through HIF-VEGF pathway, and high HIF-$1{\alpha}$ expression could be associated with lymph node metastasis and post operative poor prognosis in patients with non-small cell lung carcinoma.

The Discriminating Nature of Dopamine Transporter Image in Parkinsonism: The Competency of Dopaminergic Transporter Imaging in Differential Diagnosis of Parkinsonism: $^{123}I-FP-CIT$ SPECT Study (도파민운반체 영상의 파킨슨증 감별진단 성능: $^{123}I-FP-CIT$ SPECT 연구)

  • Kim, Bom-Sahn;Jang, Sung-June;Eo, Jae-Seon;Park, Eun-Kyung;Kim, Yu-Kyeong;Kim, Jong-Min;Lee, Won-Woo;Kim, Sang-Eun
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.4
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    • pp.272-279
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    • 2007
  • Purpose: The aim of this study was to evaluate the discriminating nature of $^{123}I-FP-CIT$ SPECT in patients with parkinsonism. Methods: $^{123}I-FP-CIT$ SPECT images acquired from the 18 normal controls; NC ($60.4{\pm}10.0$ yr) and 237 patients with parkinsonism ($65.9{\pm}9.2$ yr) were analyzed. From spatialIy normalized images, regional counts of the caudate, putamen, and occipital lobe were obtained using region of interest method. Binding potential (BP) was calculated with the ratio of specific to nonspecific binding activity at equilibrium. Additionally, the BP ratio of putamen to caudate (PCR) and asymmetric Index (ASI) were measured. Results: BPs of NC $3.37{\pm}0.57,\; 3.10{\pm}0.41,\; 3.23{\pm}0.48$ for caudate, putamen, whole striatum, respectively) had no significant difference with those of essential tremor; ET ($3.31{\pm}0.64,\; 3.06{\pm}0.61,\; 3.14{\pm}0.63$) and Alzheimer's disease; AD (3.33 $\pm$0.60, 3.29$\pm$0.79, 3.31$\pm$0.70), but were higher than those of Parkinson's disease; PD (1.92$\pm$0.74, 1.39$\pm$0.68, 1.64$\pm$0.68), multiple system atrophy; MSA (2.36$\pm$1.07, 2.16$\pm$0.91, 2.26$\pm$0.96), and dementia with Lewy body; DLB (1.95$\pm$0.72, 1.64$\pm$0.65, 1.79$\pm$0.66)(p<0.005). PD had statisticalIy lower values of PER and higher values of ASI than those of NC (p<0.005). And PD had significantIy lower value of PCR, higher ASI and lower BP in the putamen and whole striatum than MSA (p<0.05). Conclusion: Dopamine transporter image of $^{123}I-FP-CIT$ SPECT was a good value in differential diagnosis of parkinsonism.

Variations in Ammonium Removal Rate with Tidal State in the Macrotidal Han River Estuary: Potential Role of Nitrification (한강기수역에서의 암모늄 제거율 변화 및 질산화의 잠재적 역할)

  • Hyun, Jung-Ho;Chung, Kyung-Ho;Park, Yong-Chul;Choi, Joong-Ki
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.4 no.1
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    • pp.33-39
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    • 1999
  • In order to understand the importance of tidal action and $NH_4{^+}$ -nitrification in the removal of dissolved oxygen (DO) and $NH_4{^+}$, concentrations of DO, $NH_4{^+}$, $NO_2{^-}$ and $NO_3{^-}$ were measured with time for water samples collected at different tidal state in the eutrophic macrotidal Han River estuary. Field measurements indicated that most environmental parameters, except for the water temperature and DO concentration, were tightly controlled by the eutrophic freshwater runoff and large-scale tidal action. Dark incubation of the water sample at $25^{\circ}C$ showed that the removal rates of DO and $NH_4{^+}$ in high tide sample were 2.76 ${\mu}M\;O_2\;d^{-1}$ and 1.76 ${\mu}M\;N\;d^{-1}$ respectively, and increased to 5.66 ${\mu}M\;O_2\;d^{-1}$ and 3.36 ${\mu}M\;N\;d^{-1}$ respectively, in low tide sample. These changes indicated that microbial degradation and uptake of organic matter and inorganic nutrients were more active during low tide. $NH_4{^+}$-nitrification responsible for total DO removal in low tide (23.81%) and $NH_4{^+}$ turnover rates due to $NH_4{^+}$-nitrification in low tide (0.18 $d^{-1}$) were approximately 3.7 times and 3 times, respectively, higher than those in high tide. These results indicated that $NH_4{^+}$ -nitrifying bacteria introduced into the Han River estuary during low tide played a significant role in the removal of DO and $NH_4{^+}$. The decreasing removal rates in DO and $NH_4{^+}$ with the increasing tidal level seemed to be associated with the salinity impact on the halophobic freshwater $NH_4{^+}$-nitrifying bacteria. The results implied that anthropogenic $NH_4{^+}$ sources should be treated prior to the freshwater runoff into the estuary for the effective control of $NH_4{^+}$ in the Han River estuary. These results also suggest that parallel ecological studies on the chemoautotrophic nitrifying bacteria are essential for the elucidation of nitrogen cycles in the eutrophic Han River estuary.

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Sensitivity of Simulated Water Temperature to Vertical Mixing Scheme and Water Turbidity in the Yellow Sea (수직 혼합 모수화 기법과 탁도에 따른 황해 수온 민감도 실험)

  • Kwak, Myeong-Taek;Seo, Gwang-Ho;Choi, Byoung-Ju;Kim, Chang-Sin;Cho, Yang-Ki
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.18 no.3
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    • pp.111-121
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    • 2013
  • Accurate prediction of sea water temperature has been emphasized to make precise local weather forecast and to understand change of ecosystem. The Yellow Sea, which has turbid water and strong tidal current, is an unique shallow marginal sea. It is essential to include the effects of the turbidity and the strong tidal mixing for the realistic simulation of temperature distribution in the Yellow Sea. Evaluation of ocean circulation model response to vertical mixing scheme and turbidity is primary objective of this study. Three-dimensional ocean circulation model(Regional Ocean Modeling System) was used to perform numerical simulations. Mellor- Yamada level 2.5 closure (M-Y) and K-Profile Parameterization (KPP) scheme were selected for vertical mixing parameterization in this study. Effect of Jerlov water type 1, 3 and 5 was also evaluated. The simulated temperature distribution was compared with the observed data by National Fisheries Research and Development Institute to estimate model's response to turbidity and vertical mixing schemes in the Yellow Sea. Simulations with M-Y vertical mixing scheme produced relatively stronger vertical mixing and warmer bottom temperature than the observation. KPP scheme produced weaker vertical mixing and did not well reproduce tidal mixing front along the coast. However, KPP scheme keeps bottom temperature closer to the observation. Consequently, numerical ocean circulation simulations with M-Y vertical mixing scheme tends to produce well mixed vertical temperature structure and that with KPP vertical mixing scheme tends to make stratified vertical temperature structure. When Jerlov water type is higher, sea surface temperature is high and sea bottom temperature is low because downward shortwave radiation is almost absorbed near the sea surface.

New Approaches for Overcoming Current Issues of Plasma Sputtering Process During Organic-electronics Device Fabrication: Plasma Damage Free and Room Temperature Process for High Quality Metal Oxide Thin Film

  • Hong, Mun-Pyo
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.100-101
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    • 2012
  • The plasma damage free and room temperature processedthin film deposition technology is essential for realization of various next generation organic microelectronic devices such as flexible AMOLED display, flexible OLED lighting, and organic photovoltaic cells because characteristics of fragile organic materials in the plasma process and low glass transition temperatures (Tg) of polymer substrate. In case of directly deposition of metal oxide thin films (including transparent conductive oxide (TCO) and amorphous oxide semiconductor (AOS)) on the organic layers, plasma damages against to the organic materials is fatal. This damage is believed to be originated mainly from high energy energetic particles during the sputtering process such as negative oxygen ions, reflected neutrals by reflection of plasma background gas at the target surface, sputtered atoms, bulk plasma ions, and secondary electrons. To solve this problem, we developed the NBAS (Neutral Beam Assisted Sputtering) process as a plasma damage free and room temperature processed sputtering technology. As a result, electro-optical properties of NBAS processed ITO thin film showed resistivity of $4.0{\times}10^{-4}{\Omega}{\cdot}m$ and high transmittance (>90% at 550 nm) with nano- crystalline structure at room temperature process. Furthermore, in the experiment result of directly deposition of TCO top anode on the inverted structure OLED cell, it is verified that NBAS TCO deposition process does not damages to the underlying organic layers. In case of deposition of transparent conductive oxide (TCO) thin film on the plastic polymer substrate, the room temperature processed sputtering coating of high quality TCO thin film is required. During the sputtering process with higher density plasma, the energetic particles contribute self supplying of activation & crystallization energy without any additional heating and post-annealing and forminga high quality TCO thin film. However, negative oxygen ions which generated from sputteringtarget surface by electron attachment are accelerated to high energy by induced cathode self-bias. Thus the high energy negative oxygen ions can lead to critical physical bombardment damages to forming oxide thin film and this effect does not recover in room temperature process without post thermal annealing. To salve the inherent limitation of plasma sputtering, we have been developed the Magnetic Field Shielded Sputtering (MFSS) process as the high quality oxide thin film deposition process at room temperature. The MFSS process is effectively eliminate or suppress the negative oxygen ions bombardment damage by the plasma limiter which composed permanent magnet array. As a result, electro-optical properties of MFSS processed ITO thin film (resistivity $3.9{\times}10^{-4}{\Omega}{\cdot}cm$, transmittance 95% at 550 nm) have approachedthose of a high temperature DC magnetron sputtering (DMS) ITO thin film were. Also, AOS (a-IGZO) TFTs fabricated by MFSS process without higher temperature post annealing showed very comparable electrical performance with those by DMS process with $400^{\circ}C$ post annealing. They are important to note that the bombardment of a negative oxygen ion which is accelerated by dc self-bias during rf sputtering could degrade the electrical performance of ITO electrodes and a-IGZO TFTs. Finally, we found that reduction of damage from the high energy negative oxygen ions bombardment drives improvement of crystalline structure in the ITO thin film and suppression of the sub-gab states in a-IGZO semiconductor thin film. For realization of organic flexible electronic devices based on plastic substrates, gas barrier coatings are required to prevent the permeation of water and oxygen because organic materials are highly susceptible to water and oxygen. In particular, high efficiency flexible AMOLEDs needs an extremely low water vapor transition rate (WVTR) of $1{\times}10^{-6}gm^{-2}day^{-1}$. The key factor in high quality inorganic gas barrier formation for achieving the very low WVTR required (under ${\sim}10^{-6}gm^{-2}day^{-1}$) is the suppression of nano-sized defect sites and gas diffusion pathways among the grain boundaries. For formation of high quality single inorganic gas barrier layer, we developed high density nano-structured Al2O3 single gas barrier layer usinga NBAS process. The NBAS process can continuously change crystalline structures from an amorphous phase to a nano- crystalline phase with various grain sizes in a single inorganic thin film. As a result, the water vapor transmission rates (WVTR) of the NBAS processed $Al_2O_3$ gas barrier film have improved order of magnitude compared with that of conventional $Al_2O_3$ layers made by the RF magnetron sputteringprocess under the same sputtering conditions; the WVTR of the NBAS processed $Al_2O_3$ gas barrier film was about $5{\times}10^{-6}g/m^2/day$ by just single layer.

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Real-Time RT-PCR for Validation of Reovirus Type 3 Safety During the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals (세포배양 유래 생물의약품 생산 공정에서 Reovirus Type 3 안전성 검증을 위한 Real-Time RT-PCR)

  • Lee, Dong-Hyuck;Jeong, Hyo-Sun;Kim, Tae-Eun;Oh, Seon-Hwan;Lee, Jung-Suk;Kim, In-Seop
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.228-236
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    • 2008
  • Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to Reovirus type 3 (Reo-3), and there are several reports of Reo-3 contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the Reo-3 safety, a real-time RT-PCR method was developed for quantitative detection of Reo-3 in cell lines, raw materials, manufacturing processes, and final products as well as Reo-3 clearance validation. Specific primers for amplification of Reo-3 RNA was selected, and Reo-3 RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $3.2{\times}10^0\;TCID_{50}/ml$. The real-time RT-PCR method was proven to be reproducible and very specific to Reo-3. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with Reo-3. Reo-3 RNA could be quantified in CHO cell as well as culture supernatant. When the real-time RT-PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of Reo-3.