• Title/Summary/Keyword: Bio safety level

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Intracellular CD154 Expression Reflects Antigen-specific $CD8^+\;T$ Cells but Shows Less Sensitivity than Intracellular Cytokine and MHC Tetramer Staining

  • Han, Young-Woo;Aleyas, Abi G.;George, Junu A.;Yoon, Hyun-A;Lee, John-Hwa;Kim, Byung-Sam;Eo, Seong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1955-1964
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    • 2007
  • A recent report showed that analysis of CD154 expression in the presence of the secretion inhibitor Brefeldin A (Bref A) could be used to assess the entire repertoire of antigen-specific $CD4^+\;T$ helper cells. However, the capacity of intracellular CD154 expression to identify antigen-specific $CD8^+\;T$ cells has yet to be investigated. In this study, we compared the ability of intracellular CD154 expression to assess antigen-specific $CD8^+\;T$ cells with that of accepted standard assays, namely intracellular cytokine IFN-${\gamma}$ staining (ICS) and MHC class I tetramer staining. The detection of intracellular CD154 molecules in the presence of Bref A reflected the kinetic trend of antigen-specific $CD8^+\;T$ cell number, but unfortunately showed less sensitivity than ICS and tetramer staining. However, ICS levels peaked and saturated 8 h after antigenic stimulation in the presence of Bref A and then declined, whereas intracellular CD154 expression peaked by 8 h and maintained the saturated level up to 24 h post-stimulation. Moreover, intracellular CD154 expression in antigen-specific $CD8^+\;T$ cells developed in the absence of $CD4^+\;T$ cells changed little, whereas the number of IFN-${\gamma}$-producing $CD8^+\;T$ cells decreased abruptly. These results suggest that intracellular CD154 could aid the assessment of antigen-specific $CD8^+\;T$ cells, but does not have as much ability to identify heterogeneous $CD4^+\;T$ helper cells. Therefore, the combined analytical techniques of ICS and tetramer staining together with intracellular CD154 assays may be able to provide useful information on the accurate phenotype and functionality of antigen-specific $CD8^+\;T$ cells.

CD8+ T Cell-mediated Immunity Induced by Heterologous Prime-boost Vaccination Based on DNA Vaccine and Recombinant Vaccinia Virus Expressing Epitope (Epitope발현 DNA Vaccine과 Recombinant Vaccinia Virus를 이용한 Heterologous Prime-boost Vaccination에 의하여 유도되는 CD8+ T 세포 매개성 면역)

  • Park, Seong-Ok;Yoon, Hyun-A;Aleyas, Abi George;Lee, John-Hwa;Chae, Joon-Seok;Eo, Seong-Kug
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.89-98
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    • 2005
  • Background: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. Methods: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, $gB_{498-505}$ DNA) and recombinant vaccinia virus $(vvgB_{498-505})$ expressing epitope $gB_{498-505}$ (SSIEF ARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with $gB_{498-505}$ epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. Results: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and $vvgB_{498-505}$, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with $vvgB_{498-505}$ as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly $gB_{498-505}$ DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and $vvgB_{498-505}$. Of particular interest CD8+ T cell-mediated immunity was optimally induced when $vvgB_{498-505}$ was used to prime and gB DNA was used as alternative boost. Especially CD7+ T cell responses induced by such protocol was longer lasted than other protocols. Conclusion: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.

Effects of Alpha-G Rutin Supplementation in Sperm Freezing Extender on Dog Sperm Cryopreservation

  • Park, Sang-Hyoun;Jeon, Yubyeol;Talha, Nabeel Abdelbagi Hamad;Yu, Il-Jeoung
    • Journal of Veterinary Clinics
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    • v.36 no.5
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    • pp.259-265
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    • 2019
  • This study was designed to investigate the effects of alpha-glucosyl rutin (G-rutin) and its comparative effects with other antioxidants (glutathione: GSH, catalase: CATA and beta-mercaptoethanol : ${\beta}ME$) on dog sperm freezing. In the first experiment (E1), the spermatozoa were diluted in freezing extender supplemented with 0 (control), 0.001, 0.01, or 0.1% G-rutin and frozen using liquid nitrogen ($LN_2$). The progressive motility, reactive oxygen species (ROS) level and apoptosis of spermatozoa were assessed after sperm thawing at $37^{\circ}C$ for 25 sec. In the second experiment (E2), 0.1% G-rutin group was compared with 10 mM ${\beta}-ME$, $5{\mu}M$ GSH and $50{\mu}M$ CATA groups by assaying progressive motility, viability and gene expression of Bcl-2 and SMCP after sperm freezing and thawing. In E1, 0.1% G-rutin group showed higher (P < 0.05) post-thaw progressive motility and lower (P < 0.05) ROS levels. In E2, the expressions of SMCP in G-rutin group were higher (P < 0.05) than in CATA group while Bcl-2 expression of G-rutin group was higher (P < 0.05) than ${\beta}-ME$ and CATA groups. However, there were no significant differences in progressive motility and viability. Therefore, we suggest that G-rutin can be used as a potentially antioxidative supplement in dog sperm freezing extender on the basis of gene expression related to motility and apoptosis as well as ROS level.

Trace level analysis of Pb in plasma by inductively coupled plasma/mass spectrometry (유도결합플라즈마 질량분석법을 이용한 혈장 중 극미량 납 분석)

  • Lee, Sung-Bae;Yang, Jeong-Sun;Choi, Sung-Bong;Shin, Ho-Sang
    • Analytical Science and Technology
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    • v.25 no.3
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    • pp.190-196
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    • 2012
  • The human exposure of lead has usually detected the amount of lead in the whole blood, however, this method has a shortcoming to give the information on the short-term exposure to lead. In that sense, it is desirable to estimates the level of lead in plasma to draw the chronic bio-marker of lead exposure even though it is difficult to measure lead of several ng/L. An inductively coupled plasma-mass spectrometry (ICP-MS) method was developed for determining lead in plasma as the chronic bio-marker of lead of workers. To minimize the contamination of lead from the environment, we constructed class 1,000 clean room and compared the amount of floating dust before and after the operation of the clean room. The limit of detection (LOD) and the limit of quantification (LOQ) of lead in fetal bovine serum were 4.3 ng/L and 12.2 ng/L by NIOSH method (statistical calculation method) and 7.0 ng/L and 22.1 ng/L by signal/noise ratio, respectively. The accuracy was in a range of 92.3-101.3%, and the precision of the assay was less than 4% in the samples spiked in the concentration of 20 ng/L and 2,000 ng/L. The method was simple, reproducible and sensitive enough to permit reliable analysis of lead to the ng/L level in plasma and/or serum. The method was also useful for the biological monitoring of chronic exposure to lead.

Analysis of Pesticides in Herbal Medicine by QuEChERS and GC-MS/MS (QuEChERS 전처리와 GC-MS/MS를 활용한 생약의 잔류농약 분석 가능성 연구)

  • Seo, Go Eun;Kim, A Young;Pyo, Byoung Sik;Lee, Kyoung in
    • Korean Journal of Pharmacognosy
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    • v.51 no.3
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    • pp.207-216
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    • 2020
  • The analysis method for qualitative analysis of the screening method that can be performed prior to the quantitative analysis of individual pesticide was reviewed in order to meet the safety standards that are being strengthened in the field of pesticide residue testing of herbal medicines. Among the residual pesticides presented in the Korean Pharmacopoeia, 56 pesticides, excluding 15 pesticides that need to be individually analyzed, were selected for analysis using QuEChERS preprocessing and GC-MS/MS, which are used in the existing agricultural products field. For each pesticide, the detection limit level of 0.001-0.005 mg/kg and the quantitative limit level of 0.002-0.017 mg/kg were confirmed. In the recovery test in which the standard was treated at a concentration of 0.02 mg/kg, it was confirmed that the proportion of pesticides satisfying the recovery of 70-120% was 85.7-96.4% for each herbal medicine, so it was confirmed that it was a level that could be reviewed by the screening method.

Study on quantitative danger evaluation technique about frequency and strength of occurrences through accident analysis of laboratories (실험실의 사고분석을 통한 발생빈도 및 강도의 정량적 위험성 평가 기법에 관한 연구)

  • Kim, Jong-In;Rie, Dong-Ho;Choi, Soon-Young
    • Proceedings of the Safety Management and Science Conference
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    • 2008.11a
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    • pp.623-631
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    • 2008
  • In this study, with the objects of 173 institutions of university and research institutions, investigation was carried out through visit investigation and questionnaire investigation. Frequency and strength of occurrences was acquired mainly through universities and research institutions which have reported occurrences of accidents to the Ministry of Education, Science, and Technology. Quantitative danger evaluation was conducted through frequency and strength of occurrences. Case study was carried out by selecting bio science laboratory of university and as the result of this study, degree of danger among the inspection objects of bio science laboratories, laboratory work place was evaluated as the most dangerous with the scores of 155.00 and as the result of average danger level, compression gas cylinder was evaluated as the most dangerous with the scores of 6.4000.

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Wavelet Power Spectrum Estimation for High-resolution Terahertz Time-domain Spectroscopy

  • Kim, Young-Chan;Jin, Kyung-Hwan;Ye, Jong-Chul;Ahn, Jae-Wook;Yee, Dae-Su
    • Journal of the Optical Society of Korea
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    • v.15 no.1
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    • pp.103-108
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    • 2011
  • Recently reported asynchronous-optical-sampling terahertz (THz) time-domain spectroscopy enables high-resolution spectroscopy due to a long time-delay window. However, a long-lasting tail signal following the main pulse is often measured in a time-domain waveform, resulting in spectral fluctuation above a background noise level on a high-resolution THz amplitude spectrum. Here, we adopt the wavelet power spectrum estimation technique (WPSET) to effectively remove the spectral fluctuation without sacrificing spectral features. Effectiveness of the WPSET is verified by investigating a transmission spectrum of water vapor.

An Analytical Study on the Prediction of Indoor Air Quality and the Reduction of Air Conditioning Load in Bio Safety Level 3 Laboratory (생물안전 3등급시설의 공기환경 예측 및 공조부하 절감에 대한 해석)

  • Hong, Jin Kwan;Park, Hyun Jin
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.24 no.11
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    • pp.813-822
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    • 2012
  • In this study, the multizone simulation for biosafety of BSL3 lab. and energy simulation are carried out simultaneously by using linked model of CONTAM and TRNSYS. In BSL3 lab., annual energy consumption is approximately five to ten times more than the magnitude of the office building. This is because required air change rate is extremely large and it is difficult to maintain room pressure difference efficiently. To maintain pressure difference between laboratory rooms through sealing condition of doors and proper airflow control is significant. In this study, to predict indoor environment of the BSL3 lab.(Influenza A research lab.), the multizone simulation for four kinds of biohazard scenario is also performed as part of risk assessment. Multizone and energy simulation results by using linked model show that these approaches are used as a tool for the energy efficient design and operation method for the safer BSL3 lab. facilities.

A study on establishment of bio-hazard analysis and critical control point for pork slaughter house (돼지도축장의 생물학적 위해요소에 대한 중요관리점 설정)

  • 옥천석;정지영;송은아;이성식
    • Korean Journal of Veterinary Service
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    • v.24 no.2
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    • pp.117-126
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    • 2001
  • To ensure the safety of domestic livestock products, the government made it obligatory to enforce the hazard analysis critical control point(HACCP) in all domestic slaughterhouses. Under the HACCP, most of the hazards generated in slaughterhouses are bio-hazards, especially pathogenic bacteria. In order to reduce to the pathogenic bacteria, critical control point (CCP) is established and controled in the process of slaughter. A study was carried out to measure the level of bacterial contamination of swine carcass in 6 slaughterhouses selected. As a result, the aerobic plate counts(cfu/$\textrm{cm}^2$) of all samples was $10^2$-10 in average, except slaughterhouse C. The level of the aerobic plate counts on the surface of lower loin in slaughterhouse C was $10^4$ and it was considered that slaughterhouse C should set the process of manual transport of carcass as the CCP. Escherichia coli level was the highest in middle line cut surface. Especially, E coli level of slaughterhouses C and D were about 6.5- and 3.0-fold higher than that of other 4 slaughterhouses. Thus, it was considered that the slaughterhouses C and D should set the process of the entrails treatment as the CCP. The air contamination was measured at two point in a slaughterhouse. The air contamination level was 4-13 times higher than that of the standard Japanese slaughterhouses.

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The Removal Efficacy of Heavy Metals and Total Petroleum Hydrocarbons from Contaminated Soils by Integrated Bio-phytoremediation

  • Lai, Wen-Liang;Lee, Fang-Yin;Chen, Colin S.;Hseu, Zeng-Yei;Kuo, Yau-Lun
    • Journal of Soil and Groundwater Environment
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    • v.19 no.5
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    • pp.35-44
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    • 2014
  • In this study, the bio-phytoremediation and phytoremediation technologies were applied to the soils contaminated with total petroleum hydrocarbons (TPH) and heavy metals to evaluate the remediation efficacy from May 2012 to December 2013. Poplar (Populus bonatii Levl.) and Sun Hemp (Crotalaria juncea L.) were selected and planted in phytoremediation practice. These plants were also utilized in the bio-phytoremediation practice, with the addition of earthworm (Eisenia fetida) and petroleum-degrading bacteria (Pseudomonos sp. NKNU01). Furthermore, physiological characteristics, such as photosynthesis rate and maximal photochemical yield, of all testing plants were also measured in order to assess their health conditions and tolerance levels in adverse environment. After 20 months of remedial practice, the results showed that bio-phytoremediation practice had a higher rate of TPH removal efficacy at 30-60 cm depth soil than that of phytoremediation. However, inconsistent results were discovered while analyzing the soil at 100 cm depth. The study also showed that the removal efficiency of heavy metals was lower than that of TPH after remediation treatment. The results from test field tissue sample analysis revealed that more Zinc than Chromium was absorbed and accumulated by the tested plants. Plant height measurements of Poplar and Sun Hemp showed that there were insignificant differences of growth between the plants in remediation plots and those in the control plot. Physiological data of Poplar also suggested it has higher tolerance level toward the contaminated soils. These results indicated that the two testing plants were healthy and suitable for this remediation study.