• Title/Summary/Keyword: Bio safety level

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A Study on Verification for the Design of Bio Safety Level 3 Laboratory by using Multi-zone Simulation (멀티죤 시뮬레이션을 이용한 생물안전 3등급(BSL3)시설의 설계 검증에 관한 연구)

  • Lee, Hyun-Woo;Choi, Sang-Gon;Hong, Jin-Kwan
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.21 no.12
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    • pp.671-677
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    • 2009
  • In Korea, since the implementation of the LMO Law, the interest of biosafety level 3(BL3) lab. is increasing. In this study, using CONTAM which is applying multizone modelling, the multizone simulation for design verification of BL3 lab. is performed. In BL3 lab., because required air change rate is greater than general estimated air-conditioning load and it is difficult to maintain room pressure difference efficiently, to maintain pressure difference between laboratory rooms is important through sealing condition of doors and proper airflow control of laboratory rooms. In this study, about BL3 lab,(M. tuberculosis research lab.), the multizone simulation for four kind of biohazard scenarios is carried out in the case of unexpected spread of contaminants in the laboratory room, anteroom, corridor and inside of BSC. Multizone simulation results show that these approach methods are used as a tool for the design and verification of BL3 lab.

Distinct Humoral and Cellular Immunity Induced by Alternating Prime-boost Vaccination Using Plasmid DNA and Live Viral Vector Vaccines Expressing the E Protein of Dengue Virus Type 2

  • George, Junu A.;Eo, Seong-Kug
    • IMMUNE NETWORK
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    • v.11 no.5
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    • pp.268-280
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    • 2011
  • Background: Dengue virus, which belongs to the Flavivirus genus of the Flaviviridae family, causes fatal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) with infection risk of 2.5 billion people worldwide. However, approved vaccines are still not available. Here, we explored the immune responses induced by alternating prime-boost vaccination using DNA vaccine, adenovirus, and vaccinia virus expressing E protein of dengue virus type 2 (DenV2). Methods: Following immunization with DNA vaccine (pDE), adenovirus (rAd-E), and/or vaccinia virus (VV-E) expressing E protein, E protein-specific IgG and its isotypes were determined by conventional ELISA. Intracellular CD154 and cytokine staining was used for enumerating CD4+ T cells specific for E protein. E protein-specific CD8+ T cell responses were evaluated by in vivo CTL killing activity and intracellular IFN-${\gamma}$ staining. Results: Among three constructs, VV-E induced the most potent IgG responses, Th1-type cytokine production by stimulated CD4+ T cells, and the CD8+ T cell response. Furthermore, when the three constructs were used for alternating prime-boost vaccination, the results revealed a different pattern of CD4+ and CD8+ T cell responses. i) Priming with VV-E induced higher E-specific IgG level but it was decreased rapidly. ii) Strong CD8+ T cell responses specific for E protein were induced when VV-E was used for the priming step, and such CD8+ T cell responses were significantly boosted with pDE. iii) Priming with rAd-E induced stronger CD4+ T cell responses which subsequently boosted with pDE to a greater extent than VV-E and rAd-E. Conclusion: These results indicate that priming with live viral vector vaccines could induce different patterns of E protein-specific CD4+ and CD8+ T cell responses which were significantly enhanced by booster vaccination with the DNA vaccine. Therefore, our observation will provide valuable information for the establishment of optimal prime-boost vaccination against DenV.

Effects of Green Tea (Camellia sinensis) Extract Supplementation at Different Dilution Steps on Boar Sperm Cryopreservation and in vitro Fertilization

  • Park, Sang-Hyoun;Jeon, Yubyeol;Yu, Il-Jeoung
    • Journal of Veterinary Clinics
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    • v.35 no.2
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    • pp.39-45
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    • 2018
  • We evaluated the effects of green tea extract (GTE) supplementation at different dilution steps on boar sperm freezing and in vitro fertilization. Sperm intracellular hydrogen peroxide ($H_2O_2$), motility, viability, acrosome integrity and morphology were determined. In addition, sperm IVF parameters (penetration and monospermy) and glutathione (GSH) levels of presumptive zygotes (PZs) were evaluated. Semen was diluted in lactose egg yolk (LEY) and cooled at $5^{\circ}C$ for 3 h (first dilution step) and then diluted in LEY with 9% glycerol and maintained at $5^{\circ}C$ for 30 min (second dilution step). Four experimental groups were compared: first and second dilution steps without GTE (control), first dilution step with GTE (Step 1), second dilution step with GTE (Step 2) and first and second dilution step with GTE (Step 1+2). The spermatozoa were frozen in nitrogen vapor. Higher sperm motility, viability and acrosome integrity after thawing were observed in Step 1, Step 2 and Step 1+2 groups compared with the control (P < 0.05). Lower $H_2O_2$ level was observed in Step 1+2 compared with control and Step 1 (P < 0.05). For IVF, matured oocytes were co-cultured with spermatozoa frozen according to the experimental groups. GSH levels of PZs were significantly higher in Step 2 and Step 1+2 than in control and Step 1 (P < 0.05) without a significant difference in IVF parameters. In conclusion, supplementation with GTE in both first and second dilution steps during the freezing process resulted in better boar sperm cryopreservation and might be beneficial for further embryo development.

Introduction to the Technology, Applications, Products, Markets, R&D, and Perspectives of Nanofoods in the Food Industry

  • Kim, Dong-Myong;Lee, Gee-Dong
    • Preventive Nutrition and Food Science
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    • v.11 no.4
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    • pp.348-357
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    • 2006
  • Nano is a unit that designates a billionth; accordingly nanotechnology could be described as the study and applications of the unique characteristics and phenomena of nanometer size materials. Applications of nanotechnology fall into two categories (one is top-down and the other is bottom-up). Currently, most products are the results of the top-down approach. Nanofoods have distinct functional characteristics stemming from the size, mass, chemical combinations, electrolytic features, magnetic properties of food sources at the nano level and which can be applied for safe absorption and delivery into the body. The greatest advantage of nanofood is that it permits the efficient use of small quantities of nutritional elements by increasing digestive absorption ability and by delivering natural elements without any change in their original characteristics. On the other hand, there are still unsolved problems, such as questions about safety and introduction of harmful material. The demand for new commercial food products is increasing, and commercial food producers are gradually combining nanotechnology and traditional food preparation methods. Nanofoods will improve our eating habits remarkably in the future. Tomorrow we will design nanofoods by shaping molecules and atoms. It will have a big impact on the food and food-processing industries. The future belongs to new products and new processes with the goals of customizing and personalizing consumer products. Nanotechnology is expected to be applied to not only foods themselves, but also to food packaging, production, safety, processing and storage. Also, it is believed that nanotechnology will be applied tracking finished products back to production facilities and even to specific processing equipment in those facilities. The aim of this study is the introduction of technology, applications, products, markets, R&D, and perspectives of nanofoods in the food industry.

Effects of High Dose Lysophosphatidic Acid Supplement during IVC on Preimplantation Development of Porcine Embryos

  • Jin, Minghui;Yu, Il-Jeoung;Jeon, Yubyeol
    • Journal of Embryo Transfer
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    • v.32 no.4
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    • pp.275-285
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    • 2017
  • Lysophosphatidic acid (LPA) is an important signaling molecule. Here, the effect and mechanism of LPA on the preimplantation development of porcine embryos during in vitro culture (IVC) was examined. Porcine embryos were cultured in porcine zygote medium (PZM-3) supplemented with $30{\mu}M$ LPA during different days. There was a significantly higher cleavage rate in Day 1-7 and significantly higher total cell number of blastocysts in Day 1-3 and Day 4-7. It was also found that messenger RNA (mRNA) expression level of PCNA, BCL2 and BAX in blastocysts obtained from D1-7 group were significantly higher and BCL2/BAX mRNA ratio in D1-3 group was significantly lower than control group but Day 4-7 and Day 1-7 groups were comparable with control group. Treatment with $20{\mu}M$ PLC inhibitor significantly decreased the embryo cleavage rate and blastocyst formation rate. Moreover, LPA as an activator of PLCs, enhanced the $30{\mu}M$ LPA + $20{\mu}M$ U73122 group embryo cleavage rate which similar with control group. In conclusion, the results suggest that treatment with LPA during IVC improves the porcine early embryo cleavage by activation of PLC signaling pathway and regulate the mRNA expression that contribute to total cell number of blastocysts during blastocyst formation.

Detection of Foreign Antigen-specific $CD4^+Foxp3^+$ Regulatory T Cells by MHC Class II Tetramer and Intracellular CD154 Staining

  • Choi, Jin Young;Eo, Seong Kug
    • IMMUNE NETWORK
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    • v.13 no.6
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    • pp.264-274
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    • 2013
  • The unrestricted population of $CD4^+Foxp3^+$ regulatory T (Treg) cells, which have been known to control the expression of autoimmune diseases and protective immunity to inflammatory reactions, has led to greater appreciation of functional plasticity. Detecting and/or isolating Ag-specific $CD4^+Foxp3^+$ Tregs at the single cell level are required to study their function and plasticity. In this study, we established and compared both MHC class II tetramer and intracellular CD154 staining, in order to detect $CD4^+Foxp3^+$ Treg specific for foreign Ag in acute and chronic infections with lymphocytic choriomeningitis virus (LCMV). Our results revealed that MHC class II tetramer staining showed a lower detection rate of LCMV $GP_{66-77}$-specific $CD4^+$ T cells because most of MHC class II tetramers were unbound and unstable when combined staining was performed with intracellular cytokines. In contrast, intracellular CD154 staining was revealed to be easier and simple for detecting LCMV $GP_{66-77}$-specific $CD4^+$ T cells, compared to MHC class II tetramer staining. Subsequently, we employed intracellular CD154 staining to detect LCMV $GP_{66-77}$-specific $CD4^+Foxp3^+$ Tregs using $Foxp3^{GFP}$ knock-in mouse, and found that LCMV $GP_{66-77}$-specific $CD4^+Foxp3^+$ Tregs and polyclonal $CD4^+Foxp3^+$ Tregs showed differential expansion in mice infected with LCMV Arms or Cl13 at acute (8 and 13 days pi) and chronic phases (35 days pi). Therefore, our results provide insight into the valuable use of intracellular CD154 staining to detect and characterize foreign Ag-specific $CD4^+Foxp3^+$ Treg in various models.

Effects of Green Tea Extract on Sperm Quality, Reactive Oxygen Species and Lipid Peroxidation in Long-term Liquid Preservation of Boar Spermatozoa

  • Park, Sang-Hyoun;Yu, Il-Jeoung
    • Journal of Veterinary Clinics
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    • v.33 no.6
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    • pp.356-361
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    • 2016
  • During storage, boar spermatozoa undergo several changes including diminished motility and viability and accumulated reactive oxygen species (ROS). In this study, we investigated the effects of green tea extract (GTE) supplementation in the Sui Dil extender on the sperm motility, viability, ROS and lipid peroxidation (LPO) of long-term preserved boar semen at $17^{\circ}C$. A total number of eight boars were used for this experiment. Pooled ejaculates were diluted to $20{\times}10^6sperm/ml$ in the Sui Dil extender containing 0 (control), 1, 10, 100 or 500 mg/l GTE and were preserved at $17^{\circ}C$ for 24, 72, 120 and 168 h, respectively. At each storage time, sperm motility and viability were estimated by microscopic examination and the fluorescent double stain $Fertilight^{(R)}$, respectively. Sperm ROS level and LPO were assessed using the 2', 7'-dichlorodihydrofluorescein diacetate ($H_2DCFDA$)/propidium iodide (PI) and C11-BODIPY581/591/PI with flow cytometry, respectively. Compared to that of the 500 mg group, there were higher sperm motility and viability in the 1, 10 and 100 mg GTE groups during the preservation from 24 to 168 h (p < 0.05). The ROS levels of the 10 and 100 mg groups during the 168 h preservation were lower than those of the 0, 1 and 500 mg groups (p < 0.05). There were no significant differences in LPO regardless of the preservation period or the GTE concentration. In conclusion, the optimal concentrations (10 and 100 mg/l) of GTE that led to lower ROS levels may be useful for liquid boar sperm preservation at $17^{\circ}C$ for a period of 168 h.

Immune response and efficacy of pigeon pox virus vaccine and fowl pox virus vaccine in chickens

  • Shil, Niraj Kanti;Rahman, Md Siddique;Hossain, M.T.;Islam, M.T.;Rahman, M.M.;Islam, K.B.M. Saiful;Cha, Se-Yeoun;Jang, Hyung-Kwan;Song, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.497-503
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    • 2007
  • The humoral immune response of chicken vaccinated with fowl and pigeon pox virus vaccines was determined with the protective potentiality of the two vaccines in field condition of Bangladesh. Different aged Fayoumi chicks were subjected for the study. To assess the relationship with better immune response among experimental groups, the average percentage of 'take reaction' was examined and recorded to 97.77% in group A, 93.33% in group B and 100.0% in group C. The level of immune status induced by different vaccinated group was measured by passive hemagglutination (PHA) microplate test method. The mean PHA titer levels after primary vaccination were $33.06{\pm}14.13$ in group A, $32.0{\pm}14.81$ in group B, and $33.0{\pm}13.66$ in group C. Following booster vaccination, the mean PHA titer levels in prior of challenge were increased to $55.46{\pm}14.64$ in groups A and C, and $46.93{\pm}16.52$ in group B. The recorded PHA titer levels of each group at two weeks after challenge were significantly increased to $106.66{\pm}31.22$, $93.86{\pm}33.04$ and $110.93{\pm}29.29$, respectively. The PHA titer levels after vaccination and challenge were significantly increased compared to pre-vaccination titer levels (P<0.01). Although the PHA titer levels among three groups administrated different vaccine combinations in prior of challenge were significantly varied (P<0.01), it was observed that all of the vaccinated chicks were highly protected against challenge infection.

Vaccination Studies against Fowl Pox in Chickens (닭에서 계두 예방 접종에 관한 연구)

  • Shil, N.K.;Rahman, M.S.;Paul, S.;Cha, S.Y.;Jang, H.K.;Song, H.J.
    • Korean Journal of Poultry Science
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    • v.34 no.4
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    • pp.253-257
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    • 2007
  • Fayoumi chicks were vaccinated with fowl pox virus vaccine and pigeon pox virus vaccine. The protective potentiality of the two vaccines was compared in field condition in Bangladesh. The percentage of 'take reaction' was assessed to conclude its relationship with better immune response and recorded 93.33% and 100% in birds of group B and group C, respectively. The mean passive hemagglutination (PHA) antibody titre after primary vaccination was $32{\pm}14.81$ in group B and $33{\pm}13.66$ in group C. Following booster vaccination, the mean PHA titres level at pre challenge of group B was $46.93{\pm}16.52\;and\;55.46{\pm}14.64$ in group C. The PHA titre of group B and C at two weeks post challenge recorded $93.86{\pm}33.04\;and\;110.93{\pm}29.29$, respectively. PHA titre significantly (P<0.01) increased after vaccination and post challenge compared to pre- vaccination titre. There was significant variation (p<0.01) of PHA titre at pre challenge in these groups using different vaccine combinations, but all the vaccinated birds resisted challenge infection.

The roles of PKC-δ on the regulation of insulin-like growth factor(IGF)-I and insulin-Like growth factor binding protein-3 secretion by all-trans retinoic acid in MCF-7 cell (MCF-7 cell에서 all-trans retinoic acid에 의한 insulin-like growth factor-I와 insulin-like growth factor binding protein-3 분비조절에 있어서 PKC-δ의 역할)

  • Lee, Sun-Mi;Kim, Sang-Hoon;Choi, Kwang-Soo;Kang, Chang-Won
    • Korean Journal of Veterinary Research
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    • v.46 no.2
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    • pp.97-105
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    • 2006
  • All-trans retinoic acid (AtRA) induces growth inhibition and apoptosis in a variety of tumer cells, including MCF-7 cells. Insulin-like growth factors (IGFs) system has been reported to be associated with the development of cancer. Although MCF-7 cell with AtRA is to be the major stimulus for the cell growth and apoptosis, the mechanism of insulin-like growth factor-I (IGF-I)/insulin-like growth factor binding protein-3 (IGFBP-3) system remains to be elucidated. Thus, this study was conducted to the effect of AtRA on the gene expression and level of IGF-I and IGFBP-3. In addition, we investigated the involvement of PKC-${\delta}$ on the IGF-I and IGFBP-3 secretion in MCF-7 cell. AtRA(${\geq}10^{-7}M$) decreased the IGF-1 secretion and mRNA expressions, but increased IGFBP-3 secretion and mRNA expressions in MCF-7 cells. Especially, the treatment of AtRA at 72 hours caused a significant reduction in the IGF-I secretion and mRNA expressions but increment in IGFBP-3 secretion and mRNA expressions (p < 0.05). $10^{-7}M$ AtRA activated PKC-${\delta}$ that is one among PKC-$\iota$, ${\alpha}$, ${\lambda}$ and ${\delta}$ in MCF-7 cell. Rotllerin, a PKC-${\delta}$ inhibitor, blocked AtRA-induced inhibition of the IGF-I and mRNA expressions, and increase of lGFBP-3 and mRNA expressions in MCF-7 cell. Together, AtRA inhibited the IGF-I secretion and mRNA expressions, but increased IGFBP-3 secretion and mRNA expressions in MCF-7 cell. Furthermore, AtRA-induced alteration of IGF-I, IGFBP-3 secretion, and the gene expressions were mediated via PKC-${\delta}$ activity.