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Increase of Spacer Sequence Yields Higher Dimer $(Fab-Spacer-Toxin)_{2}$ Formation

  • Yoo Mee-Hyeon;Won Jae-Seon;Lee Yong-Chan;Choe Mu-Hyeon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1097-1103
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    • 2006
  • The divalent antibody-toxins are expected to have increased binding avidities to target cells because of the two cell-binding domains. However, previous studies showed that the refolding yield of divalent antibody-toxin is very low, and it is assumed that homodimer formation of antibody-toxin is strongly interfered by the repulsion between the two large toxin domains that come close to each other during dimer formation. In this study, B3 antibody was used as a model antibody, and its Fab domain was used to construct three different kinds of Fab divalent molecules, $[B3(Fab)-toxin]_{2}$. The monomer Fab-toxin molecules were made by fusing the Fab domain of monoclonal antibody B3 to PE38, a truncated mutant form of Pseudomonas exotoxin (PE), and a connecting sequence that contained spacer amino acid sequence (G4S)n (n=l, 2, 3) was inserted between Fab and PE38. The prepared divalent molecules were $[Fab-S\;1,\;2,\;3-PE38]_{2}\;(=[Fab-SKPCIST-KAS(G_{4}S)nGGPE-PE38]_{2}\;(n=1,\;2,\;3))$, and they are derivatives of previously studied $[Fab-H2cys-PE38]_{2}\;(=[Fab-SKPCIST-KASGGPE-PE38]_{2})$. In $[Fab-Sl,\;2,\;3-PE38]_{2}$, two Fab-S1, 2, 3-PE38 monomers were covalently linked by the disulfide bond bridge made from cysteine in the -SKPCIST- sequence. The insertion of spacer amino acids after the disulfide bridge resulted in a 12-18 fold higher yield of dimer formation than previously constructed $[Fab-Hlcys-PZ38]_{2}[7]$, 3-4-fold higher than $[Fab-ext-PZ38]_{2}[25]$. These two molecules have less amino acid spacer sequence between the disulfide bridge and PE38 domain. The design of $[Fab-PE38]_{2}$ in this study gave molecules with a higher refolding yield. The results of cytotoxicity assay showed a higher cytotoxic effect of these divalent molecules than that of the monovalent scFv-PE38 molecule.

The Effects of Posture and Sleep Deprivation on Heart Rate Variability (자세와 수면 박탈이 심박 변이도에 미치는 영향)

  • Shim, Young-Woo;Yang, Dong-In;Kim, Nam-Hyun;Kim, Deok-Won
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.47 no.2
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    • pp.43-49
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    • 2010
  • Autonomic nervous system (ANS) acts as a control system functioning largely below the level of consciousness, and controls visceral functions. The activity of the ANS has been assessed by means of the heart rate variability (HRV). It has been reported that HRV is dependent on sex, age, body mass index, and smoking, etc. However, the effects of posture and sleep deprivation on HRV have rarely been reported. Objective of our work was to find out which posture is appropriate for stable HRV. We measured the number of sleep deprivation and HRV using power spectrum in six stages for 30 minutes. Increased low frequency (LF) power and high frequency (HF) power indicate enhanced sympathetic and parasympathetic activity, respectively. We determined the LF/HF ratio to minimize individual difference. It was found that sleep deprivation by awakening up subjects was affected by posture, which resulted in changes of LF/HF. Although LF/HF varied with time, it was more stable in sitting than in supine. In conclusion, we recommend sitting posture when measuring HRV because of less sleep deprivation resulting in less variation in LF/HF.

금산 인삼의 화학적 특성

  • 송석환;민일식
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2002.09a
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    • pp.148-151
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    • 2002
  • To clarify of chemical contents for the Keumsan ginseng, two attempts have been done for two year: 2 year and 4 year-ginsengs coilected from the phyllite(Ph0 and granite area(Gr), and 1 year, 2 year and 3 year-ginsengs collected from the phyllite(Ph), shale(Sh) and granite (Gr), In the first attempt, the soil contents of the Ph are high in most of elements. In the comparisons between ages of the ginseng, regardless the area, most of the elements are high in the 4 year and these trends are distinctive in the granite area. In the comparisons of the areas, 2 year-ginsengs of the Ph show high contents in the most of the elements. Comparisons between upper part(leaf and branch) and root part of the ginsengs show that in the 2 year-ginseng, the Gr is high in the root part and the Ph is high in the upper part, while in the 4 year-ginseng, most of the elements are high in the root part. Comparisons between contents of soil and ginsengs show similar in crease and decrese trends. However, the 2 year-ginsengs show clear differences between two areas while in the 4 year-ginseng, differences between the Gr and Ph is not clear. It suggest that the ginsengs absorb eligible element contents with ages, despite of clear content differences of the different soils. In the second attempt, the Gr shows high W, Pb, Th, U, Sn and B contents wlile Ph and Sh show high Ni, Cr, Co, Sc, V, As, Cu and Zn contents. In the comparisons between ages of the ginseng, regardless the area, most of the elements are high in the 2 year, especially in the Gr and Ph. In the comparisons of the areas, 2 year-ginsengs of the Gr show low contents in the most of the elements, and the S1 show higher Ni, V, Th and U contents than the Ph. 3 year-ginsengs of the Gr show low contents in the most of the elements, while the S1 show high contents in the most of elements. relative to the Ph. Comparisons between upper part and root part of the ginsengs show that in the ginseng, regardless the ages, the upper part is high. Comparisons between contents of soil and ginsengs show similar increase and decrese trends, and soil is low than the ginsengs in the most of the elements

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High Noise Margin LVDS I/O Circuits for Highly Parallel I/O Environments (다수의 병렬 입.출력 환경을 위한 높은 노이즈 마진을 갖는 LVDS I/O 회로)

  • Kim, Dong-Gu;Kim, Sam-Dong;Hwang, In-Seok
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.44 no.1
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    • pp.85-93
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    • 2007
  • This paper presents new LVDS I/O circuits with a high noise margin for use in highly parallel I/O environments. The proposed LVDS I/O includes transmitter and receiver parts. The transmitter circuits consist of a differential phase splitter and a output stage with common mode feedback(CMFB). The differential phase splitter generates a pair of differential signals which have a balanced duty cycle and $180^{\circ}$ phase difference over a wide supply voltage variation due to SSO(simultaneous switching output) noises. The CMFB output stage produces the required constant output current and maintains the required VCM(common mode voltage) within ${\pm}$0.1V tolerance without external circuits in a SSO environment. The proposed receiver circuits in this paper utilizes a three-stage structure(single-ended differential amp., common source amp., output stage) to accurately receive high-speed signals. The receiver part employs a very wide common mode input range differential amplifier(VCDA). As a result, the receiver improves the immunities for the common mode noise and for the supply voltage difference, represented by Vgdp, between the transmitter and receiver sides. Also, the receiver produces a rail-to-rail, full swing output voltage with a balanced duty cycle(50% ${\pm}$ 3%) without external circuits in a SSO environment, which enables correct data recovery. The proposed LVDS I/O circuits have been designed and simulated with 0.18um TSMC library using H-SPICE.

Chemical Characteristics in Fermented Cordycepin-enriched Cordyceps militaris (발효 동충하초의 이화학적 특성)

  • Ahn, Hee-Young;Park, Kyu-Rim;Kim, Yu-Ra;Cha, Jae-Young;Cho, Young-Su
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1032-1040
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    • 2013
  • The comparative effects of fibrinolytic, antioxidative activity and electrophoretical protein patterns with Cordyceps militaris powder (CM) and cordycepin-enriched Cordyceps militaris JLM0636 powder (CCM) and fermented cordycepin-enriched Cordyceps militaris JLM0636 powder by several microscopic organisms were investigated. In addition, nutritional materials such as mineral, protein, and fatty acids were also measured. The protein concentration was higher in CCM than that in CM. The protein concentration in fermented CCM was the highest in CCM treated by Aspergillus kawachii among the various samples. When total protein patterns of CM, CCM, and both fermented CCMs were analyzed by native- and SDS-PAGE, there were slightly varietal differences in electrophoretical protein patterns. Major minerals were K, Ca, Mg, and Zn. Major fatty acids were palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. Fibrinolytic activity was the highest in the fermented CCM by Bs treatment among the various samples. The ${\alpha},{\alpha}^{\prime}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) radical scavenging activity was slightly stronger in the CCM treated with Aspergillus kawachii among the various samples; however, these samples all exhibited relatively low levels of activity compared with the butylated hydroxytoluene (BHT). These results may provide the basic data necessary to understand the biological activities and chemical characteristics of Cordyceps militaris JLM0636 powder fermented by several microscopic organisms to develop functional foods.

A Policy-driven RFID Data Management Event Definition Language (정책기반 RFID 데이터 관리 이벤트 정의 언어)

  • Song, Ji-Hye;Kim, Kwang-Hoon
    • Journal of Internet Computing and Services
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    • v.12 no.1
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    • pp.55-70
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    • 2011
  • In this paper, we propose a policy-driven RFID data management event definition language, which is possibly applicable as a partial standard for SSI (Software System Infrastructure) Part 4 (Application Interface, 24791-4) defined by ISO/IEC JTC 1/SC 31/WG 4 (RFID for Item Management). The SSI's RFID application interface part is originally defined for providing a unified interface of the RFID middleware functionality―data management, device management, device interface and security functions. However, the current specifications are too circumstantial to be understood by the application developers who used to lack the professional and technological backgrounds of the RFID middleware functionality. As an impeccable solution, we use the concept of event-constraint policy that is not only representing semantic contents of RFID domains but also providing transparencies with higher level abstractions to RFID applications, and that is able to provide a means of specifying event-constraints for filtering a huge number of raw data caught from the associated RF readers. Conclusively, we try to embody the proposed concept by newly defining an XML-based RFID event policy definition language, which is abbreviated to rXPDL. Additionally, we expect that the specification of rXPDL proposed in the paper becomes a technological basis for the domestic as well as the international standards that are able to be extensively applied to RFID and ubiquitous sensor networks.

Distribution of the Quantum Dot Nano-particles that Penetrate Skin and Distinction of Combined Osmium Tetroxide in Electron Microscopic Analysis (피부로 침투된 양자점 나노입자의 분포와 전자현미경 분석 시 발견되는 오스뮴산 결합물과의 구분)

  • Choi, Ki-Ju;Park, Sang-Yong;Lee, Jeong-Min;Shin, Heon-Sub;Yang, Jung-Eun;Lee, Don-Gil;Mavonov, Garfurjon T.;Yi, Tae-Hoo
    • Applied Microscopy
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    • v.42 no.1
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    • pp.1-7
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    • 2012
  • The possibility of nanoparticles (NPs) in biotechnology had been discussed by biomedical investigators. Here we report to suggest a solution and problems when using electron microscopy to determine the distribution of quantum dots (QDs) nanoparticles that penetrate skin. The results of this study showed that NPs were able to penetrate stratum corneum (SC) and sebocyte via hair follicle. However, we have found artifacts such as nanoparticles that are produced from combination of free fatty acid and osmium tetroxide during specimen preparation. It is usually difficult to identify NPs. Therefore, we tried to resolve these problems by comparing the cross-correlation image pattern that are derived from the images of sample that had been processed differently. This method can contribute to more accurate interpretation and minimal errors during the analysis using quantum dot as tracer.

Protective Effects of Bifidobacterium bifidum Culture Supernatants and Intracellular Cell-Free Extracts on Human Dermal Fibroblasts against UV-B Irradiation (인간 진피섬유아세포에서 Bifidobacterium bifidum 배양액 및 추출액의 자외선B에 대한 보호 효능)

  • Gwon, Gi Yeong;Park, Gwi Gun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.7
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    • pp.801-808
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    • 2017
  • The present study investigated the protective effects of Bifidobacterium bifidum culture supernatants (BbSC) and intracellular cell-free extracts (BbICFE) on human dermal fibroblasts (HDFs) against ultraviolet-B (UV-B) irradiation. HDFs were treated with UV-B, UV-B+BbCS, and UV-B+BbICFE. Treatment of UV-B-irradiated HDFs with BbCS and BbICFE significantly increased cell viability compared to UV-B-irradiated HDFs. BbCS treatment reduced senescence in HDFs by approximately 40.0%. Moreover, sub-G1 phase was significantly reduced in BbCS- and BbICFE-treated HDFs (3.3% and 4.5%, respectively). The effect of UV-B on oxidative damage of HDFs was measured by dichlorofluorescin diacetate. Fluorescence intensity significantly increased in UV-B-irradiated HDFs. Inhibition of cellular reactive oxygen species in HDFs treated with 0.01% BbCS was the highest at 34.1%. Levels of p21 and p53 protein expression induced by UV-B irradiation were reduced by treatment with BbCS and BbICFE (47.0% and 35.6%, respectively). These results show that BbCS and BbICFE reduce UV-B-induced cellular senescence and apoptosis in HDFs. Thus, BbCS and BbICFE can be used as potential agents for protection of UV-B-induced skin cell damage.

Effect of Sugars on Cell Growth and Lipase Production by Trichosporon cutaneum (Trichosporon cutaneum의 균체생육(菌體生育) 및 Lipase 생산(生産)에 미치는 당류(糖類)의 영향)

  • Kim, Seung-Yeol
    • Korean Journal of Agricultural Science
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    • v.4 no.1
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    • pp.105-111
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    • 1977
  • Inhibitory effect of sugars on lipase production by Trichosporon cutaneum was observed in the previous study (Kim, 1972), and inhibition was distinctive by the addition of glucose, fructose, mannose, xylose and arabinose to the soybean meal medium among various carbon sources. These experiments were carried out to study the effect of sugars on cell growth and lipase production by the strain using the soybean extracts liquid medium under a shaking culture system. Changes in color and pH of the medium were caused by heat sterilization when various sugars were added. To elucidate the possible effect of these coloring matters on lipase production and cell growth: changes in pH of the culture, cell concentration and level of the enzyme activities were determined when the culture was grown for 48 hours at $30^{\circ}C$ on a reciprocal shaker. The results obtained were as follows: 1. Density of brownish color which formed during heat sterilization was varied with the variety of sugar used, ie, strong in pentose such as xylose: weak in hexose such as galactose, mannose, glucose: very weak in disaccharide such as maltose, sucrose. When the color density was stronger, decrease in pH after sterilization was marked. 2. Cell growth and lipase production was not so effect by the coloring matters as by sugars. 3. The more the cell mass of the culture, the lower the level of lipase production in the culture supernatant. 4. Among the sugars which caused the distinctive inhibition of lipase production, a slight relief of inhibition was noticed by the addition of xylose, whereas the cell growth was repressed. 5. When cell growth was better, decrease in pH of the medium was greater during cultivation.

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Basic Investigation for the Won-invasive Measurement of Blood Glucose Concentrations by Millimeter Waves (밀리미터파를 이용한 무혈 혈당 측정에 관한 기초 연구)

  • Kim Dong Kyun;Won Jong Hwa;Potapov Sergey N.;Meriakri Viacheslav V.;Chigryai Evgenii E.
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.42 no.1
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    • pp.39-45
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    • 2005
  • As a basic research for the development of a non-invasive blood glucose sensor using millimeter waves, we have presented a method for measuring the dielectric properties of high loss dielectrics, based on the reflection method, and investigated the variation of the dielectric properties of glucose-water and glucose -0.9% NaCl solutions in the 10~90 GHz range. In the proposed method, a minimal reflection condition is formed by placing a specially-chosen low-loss plane-parallel plate in front of a high-loss medium under test at a given frequency range. Using the minimal power reflection coefficient and the corresponding frequency at this condition, tile dielectric properties of the medium can be determined. The measured results on pure water have shown the adequacy of the proposed method. The measured results on glucose-water and glucose -0.9% NaCl solutions in the 10~90 GHz range showed that the variations of the dielectric properties of glucose solutions according to the change of their glucose concentration were maximum in the 30~45 GHz range. From these facts we concluded that the variation of about 3 mole/L in the glucose solutions must be distinguished With the measurement accuracies of ±0.1 dB and ±0.01 GHz.