• Title/Summary/Keyword: specific activity

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Site-specific and deletional mutagenesis for two regions of Verotoxin-2 A gene encoding enzymatically active domain (Verotoxin-2 A 유전자의 효소활성 부위에 대한 위치특이적 변이 및 결손변이유발)

  • Kim, Yong-hwan;Kim, Sang-hyun;Cha, In-ho;Kim, Kyoung-shook;Lee, Young-choon
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.541-546
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    • 1997
  • There are two conserved regions with a significantly high amino acid sequence homology among the A subunits of STX, SLTs and ricin. To produce an inactive Verotoxin-2 (VT-2), two different mutants, pE167D and pDE5A, were constructed by site-directed mutagenesis, respectively, on the basis of the previous reports that two regions lie within the active-site clefts of the A subunits of ricin and STX family. The cytotoxicity ($10^3$ $CD_{50}/ml$) of VT-2 holotoxin with E167D mutation was reduced by $10^3$-fold compared with wild-type level. In addition, VT-2 with DE5A ($Trp_{202}GlyArgIleSer_{206}$) deletion mutation showed a significantly low cytotoxicity ($10^1$ $CD_{50}/ml$), resulting in $10^5$- and $10^2$-fold reductions, respectively, compared with the wild-type and E167D mutatant. SDS-PAGE for protein samples showed a 33-kDa band corresponding to the A subunit of VT-2. These results indicate that reduction in cytotoxic activity was affected not by amount of VT-2 protein produced but by mutation.

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Development of Carbonization Technology and Application of Unutilized Wood Wastes(I) -Carbonization and It's Properties of Thinned Trees- (미이용 목질폐잔재의 탄화 이용개발(I) -수종의 간벌재 탄화와 탄화물의 특성-)

  • Kim, Byung-Ro;Kong, Seog-Woo
    • Journal of the Korean Wood Science and Technology
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    • v.27 no.2
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    • pp.70-77
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    • 1999
  • Objective of this research is to obtain fundamental data of carbonized wood wastes for soil condition, de-ordorization, absorption of water, carrier for microbial activity, and purifying agent for water quality of river. The carbonization technique and the properties of carbonized wood wastes(thinned trees) are analyzed. Proximate analysis shows the thinned wood contains 0.22-0.73% ash, 77-80% volatile matter, and 10-14% fixed carbon. The charcoal yield decreases and the shrinkage rate increases as the carbonization temperature and time increase. The charcoal yields of Larix leptolepis, Pinus rigida and Pinus densiflora are high, whereas those of Pinus koraiensis and Quercus variabilis are low. The shrinkage rate by carbonization has same trend as water removal of wood. The specific gravity after the carbonization decreases about 50% comparing to green wood. The charcoal has 0.89-4.08% ash, 6.31-13.79% volatile matter, and 73.9-83.5% fixed carbon. As the carbonization temperature and time increase, pH of charcoal increases. When the carbonization temperature is $400^{\circ}C$, pH is about 7.5. When the temperature is between 600 to $800^{\circ}C$, pH is about 10 with small difference. The water-retention capacity is not affected by the carbonization temperature and time. The water-retention capacity within 24hr is about 2.5 - 3times of sample weight, and the equivalent moisture content becomes 2-10% after 24 hr.

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Inflammation Scan Using $^{99m}Tc-HMPAO$ Labelled Leukocytes ($^{99m}Tc-HMPAO$를 이용한 자가백혈구표지 및 그를 이용한 염증병소의 스캔)

  • Yang, Woo-Jin;Chung, Soo-Kyo;Shinn, Kyung-Sub;Bahk, Yong-Whee;Kim, Hoon-Kyo
    • The Korean Journal of Nuclear Medicine
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    • v.23 no.2
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    • pp.219-223
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    • 1989
  • Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.

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Study on the Fatigue Crack Behavior by the Stress Intensity Factor and AE Parameters (응력확대계수와 음향방출 변수를 이용한 피로균열 거동 연구)

  • Yoon, Dong-Jin;Jeong, Jung-Chae;Park, Phi-Lip;Kim, Ki-Bok;Lee, Seung-Seok
    • Journal of the Korean Society for Nondestructive Testing
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    • v.20 no.5
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    • pp.412-423
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    • 2000
  • By using the relation between stress intensity factor and AE parameter, new approach method for assessing the crack length and detectability of crack was proposed. Laboratory experiment was carried out to identify AE characteristics of fatigue cracks for compact tension specimen. The relationship between a stress intensity factor and AE signals activity as well as conventional AE parameter analysis was discussed. As a result, the features of specific parameter such as the length of crack growth the AE energy, the AE peak amplitude, and the cumulative AE hits, showed the almost same trend in their increase as the number of fatigue cycle increased. From the comparisons of peak amplitude and AE energy with stress intensity factor, it was verified that the higher stress intensity factors generated AE signals with higher peak amplitude and a larger number of AE counts. If we can get more reliable database for the relation between AE parameters and stress intensity factor, this approach will provide a good information for evaluating both the existence of crack and the minimum detectable size of crack.

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Design and Implementation of Process Management Model applying Agent Technology (헬스케어 홈 서비스를 위한 데이터베이스 및 응용 서비스 구현)

  • Lee, Chung-Sub;Jeong, Chang-Won;Joo, Su-Chong
    • Journal of Internet Computing and Services
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    • v.8 no.1
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    • pp.57-70
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    • 2007
  • This paper is to construct a healthcare database using information obtained from healthcare home environments, and use this one for healthcare home services, Especially, our researching focus in this paper is how to design healthcare database scheme and how to use this constructed database on the Framework for Supporting Healthcare Integrated Service(FSHIS) we developed previously. Healthcare information is designed to database schemes in accordance to the specific save types of the data collected from various typed-sensors. The healthcare database constructed by using this information for the purpose of healthcare home services is divided into the base information with real schemes and the context based information with view schemes. Firstly, the base information includes low data obtained from physical sensors relevant to locations, healths, environments, and the personnel healthy profiles. The other is the context based information that is produced and fused by using the based information. This context based information might be got via various view schemes according to healthcare application services. Finally, for verifying the practical use of healthcare database constructed in this paper, Via interconnecting this database to our FSHIS, we show an example of healthcare home monitoring service using information (basic and context based information), emergency call, home appliance control, and so on needed from living activity area for elderly living alone.

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A Measurement System for Color Environment-based Human Body Reaction (색채 환경 기반의 인체 반응 정보 측정 시스템)

  • Kim, Ji-Eon;Jeong, Chang-Won;Joo, Su-Chong
    • Journal of Internet Computing and Services
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    • v.17 no.2
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    • pp.59-65
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    • 2016
  • The result of analyzing the cognitive reaction due to the color environment has been applied to various filed especially in medical field. Moreover, the study about the identification of patient's condition and examination the brain activity by collecting the bio-signal based on the color environment is being actively conducted. Even though, there were a variety of experiments by convention the color environment using a light or LED color, it still has a problem that affects the psychological information. Therefore, our proposed system using a HMD (Head Mounting display) to provide a completed color environment condition. This system uses the BMS(Biomedical System) to collect the biometric information which responds to the specific color condition and the human body response information can be measured by the development the Memory and Attention test on Mobile phone. The collection of Biometric information includes electro cardiogram(ECG), respiration, oxygen saturation (Sp02), Bio-impedance, blood pressure will store in the database. In addition, we can verify the result of the human body reaction in the color environment by Memory and Attention application. By utilizing the reaction of the human body information that is collected thought the proposed system, we can analyze the correlation between the physiological information and the color environment. And we also expect that this system can apply to the medical diagnosis and treatment. For future work, we will expand the system for prediction and treatment of Alzheimer disease by analyzing the visualization data through the proposed system. We will also do evaluation on the effectiveness of the system for using in the rehabilitation program.

Roles of Prostatic Acid Phosphatase in Prostate Cancer (Prostatic acid phosphatase의 전립선 암에서의 역할)

  • Kong, Hoon-Young;Lee, Hak-Jong;Byun, Jong-Hoe
    • Journal of Life Science
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    • v.21 no.6
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    • pp.893-900
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    • 2011
  • Prostatic acid phosphatase (PAP) is one of the widely used biomarkers in the diagnosis of prostate cancer. It was initially identified in 1935 and is the most abundant phosphatase in the human prostate. PAP is a prostate-specific enzyme that is synthesized in prostate epithelial cells. It belongs to the acid phosphatase group that shows enzymatic activity in acidic conditions. PAP is abundant in prostatic fluid and is thought to have a role in fertilization and oligospermia. It also has a potential role in reducing chronic pain. But one of the most apparent functions of PAP is the dephosphorylation of macromolecules such as HER-2 and PI3P that are involved in the ERK1/2 and MAPK pathways, which in turn leads to inhibition of cell growth and tumorigenesis. Currently, clinical trials using PAP DNA vaccine are underway and FDA-approved immunotherapy using PAP is commercially available. Despite these clinically important aspects, molecular mechanisms underlying PAP regulation are not fully understood. The promoter region of PAP was reported to be regulated by NF-${\kappa}B$, TNF-${\alpha}$, IL-1, androgen and androgen receptors. Here, the features of PAP gene and protein structures together with the function, regulation and roles of PAP in prostate cancer are discussed.

Functional Mechanism of Calmodulin for Cellular Responses in Plants (식물의 세포반응에 대한 칼모듈린의 functional 작용기작 연구)

  • Cho, Eun-Kyung;Choi, Young-Ju
    • Journal of Life Science
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    • v.19 no.1
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    • pp.129-137
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    • 2009
  • Calcium ($Ca^{2+}$) plays pivotal roles as an intracellular second messenger in response to a variety of stimuli, including light, abiotic. and biotic stresses and hormones. $Ca^{2+}$ sensor is $Ca^{2+}$-binding protein known to function in transducing signals by activating specific targets and pathways. Among $Ca^{2+}$-binding proteins, calmodulin (CaM) has been well reported to regulate the activity of down-stream target proteins in plants and animals. Especially plants possess multiple CaM genes and many CaM target proteins, including unique protein kinases and transcription factors. Thus, plants are possible to perceive different signals from their surroundings and adapt to the changing environment. However, the function of most of CaM or CaM-related proteins have been remained uncharacterized and unknown. Hence, a better understanding of the function of these proteins will help in deciphering their roles in plant growth, development and response to environmental stimuli. This review focuses on $Ca^{2+}$-CaM messenger system, CaM-associated proteins and their role in responses to external stimuli of both abiotic and biotic stresses in plants.

Enhancement of Soluble Expression of CGTase in E. coli By Chaperone Molecules and Low Temperature Cultivation. (대장균에서 chaperons 분자와 저온배양에 의한 CGTase의 가용성 발현 증대)

  • 박소림;김성구;권미정;남수완
    • Journal of Life Science
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    • v.14 no.1
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    • pp.121-125
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    • 2004
  • The synergistic effect of lowered incubation temperature and CroEL/ES expression on the production of soluble form of B. macerans cyclodextrin glucanotransferase (CGTase) was studied in recombinant E. coli. pTCGTl and pGroll carrying the cgt and groEL/ES genes under the control of T7 promoter and pzt-I promoter, respectively, were co-introduced. Tetracycline (10 ng/ml) and IPTG (1 mM) were added at the early-exponential phase (2 hr) and mid-exponential phase (3 hr). Low temperature cultivation at $25^{\circ}C$ with groEL/ES expression improved the activity of CGTase by two fold, compared to $37^{\circ}C$ cultivation without chaperones. SDS-PACE analysis revealed that about 69% of CGTase in the total CGTase protein was found in the soluble fraction by overexpression of GroEL/ES and cultivation at$25^{\circ}C$, whereas 20% of CGTase was detected in the soluble fraction when E. coli was cultivated at $37^{\circ}C$ without chaperone. The amount of soluble CGTase from $25^{\circ}C$ culture with chaperone was 3.5-fold higher than that of $37^{\circ}C$ culture without chaperone. Therefore the expression of CroEL/ES and low temperature cultivation greatly enhanced the soluble production of CGTase in E. coli.

Expression of Human SOD1 and Mutant SOD1 (G93A) in E. coli and Identification of SOD1 as a Substrate of HtrA2 Serine Protease (대장균에서의 human SOD1과 mutant SOD1 (G93A) 단백질의 발현과 HtrA2의 기질 여부 확인에 관한 연구)

  • Kim, Goo-Young;Kim, Sang-Soo;Park, Hyo-Jin;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.5
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    • pp.716-722
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    • 2006
  • Superoxide dismutase (SOD) is physiologically important in regulating cellular homeostasis and apoptotic cell death, and its mutations are the cause of familial amyotrophic lateral sclerosis (FALS). Mitochondrial serine protease HtrA2 has a pro-apoptotic function and has known to be associated with neurodegenerative disorders. To investigate the relationship between genes associated with apoptotic cell death, such as HtrA2 and SOD1, we utilized the pGEX expression system to develop a simple and rapid method for purifying wild-type and ALS-associated mutant SOD1 proteins in a suitable form for biochemical studies. We purified SOD1 and SOD1 (G93A) proteins to approximately 90% purity with relatively high yields (3 mg per liter of culture). Consistent with the result in mammalian cells, SOD1 (G93A) was more insoluble than wild-type SOD1 in E. coli, indicating that research on the aggregate formation of SOD1 may be possible using this pGEX expression system in E. coli. We investigated the HtrA2 serine protease activity on SOD1 to assess the relationship between two proteins. Not only wild-type SOD1 but also ALS-associated mutant SOD1 (G93A) were cleaved by HtrA2, resulting in the production of the 19 kDa and 21 kDa fragments that were specific for anti-SOD1 antibody. Using protein gel electrophoresis and immunoblot assay, we compared the relative molecular masses of thrombin-cleaved GST-SOD1 and HtrA2-cleaved SOD1 fragments and can predict that the HtrA2-cleavage sites within SOD1 are the peptide bonds between leucine 9-lysine 10 (L9-K10) and glutamine 23-lysine 24 (Q23-K24). Our study indicates that SOD1 is one of the substrate for HtrA2, suggesting that both HtrA2 and SOD1 may be important for modulating the HtrA2-SOD1-mediated apopotic cell death that is associated with the pathogenesis of neurodegenerative disorder.