• Title/Summary/Keyword: Ultraviolet inactivation

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Integration of a target gene into chromosomal genome of BF-2 cells using UV-inactivated snakehead retrovirus (SnRV)

  • Kwon, Se-Ryun;Nishizawa, Toyohiko;Yoshimizu, Mamoru
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.375-382
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    • 2009
  • Integration and expression of a target gene into chromosomal genomes of host cell by retrovirus mediated gene transfer system usually require complicate and laborious procedures. In the present study, we investigate a simple method to integrate a target gene into genome of BF-2 cells using ultraviolet (UV)-inactivated snakehead retrovirus (SnRV), a fish retrovirus. First of all, an optimization of transfection condition was determined with BF-2 cells using Lipofectamine 2000 and Transome. Using 0.5 $\mu\ell$ Lipofectamine 2000 resulted in 33.8, 40.6 and 40.2% of transfection efficacy with high survival rate (minimum 80%) in 0.5, 1 and 2 $\mu{g}$ DNA, respectively, and those of Transome were all less than 5%. It was confirmed that UV-treatment for 5 min was enough to inactivate infectivity of SnRV. Next, a cassette composed of GFP (green fluorescent protein) gene flanked by LTR (long terminal repeats) sequences derived from SnRV was constructed and transfected into BF-2 cells followed by treatment with UV-inactivated SnRV for optimization of integration and expression of the cassette gene. As the results, the fluorescence was expressed in BF-2 cells treated with UV-inactivated SnRV 3 and 5 times, while there was no expression in BF-2 cells with once and non treatment. Accordingly, it was confirmed that GFP gene was integrated into chromosomal genome of BF-2 cells with UV-inactivated SnRV.

Enhancement of Nitric Oxide with nonthermal plasma jet and its effect on Escherichia coli inactivation and various type of cancer cell

  • Shaw, Priyanka;Kumar, Naresh;Attri, Pankaj;Choi, Eun Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.230.2-230.2
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    • 2016
  • A new approach for antimicrobial is based on the overproduction of reactive nitrogen species (RNS), especially; nitric oxide (NO) and peroxinitrite ($ONOO^-$-) are important factors to deactivate the bacteria. Recently, non-thermal atmospheric pressure plasma jet (APPJ) has been frequently used in the field of microbial sterilization through the generation of different kinds of RNS/ROS species. However, in previous study we showed APPJ has combine effects ROS/RNS on bacterial sterilization. It is not still clear whether this bacterial killing effect has been done through ROS or RNS. We need to further investigate separate effect of ROS and RNS on bacterial sterilization. Hence, in this work, we have enhanced NO production, especially; by applying a 1% of HNO3 vapour to the N2 based APPJ. In comparison with nitrogen plasma with inclusion of water vapour plasma, it has been shown that nitrogen plasma with inclusion of 1% of HNO3 vapour has higher efficiency in killing the E. coli and different type of cancer cell through the high production of NO. We also investigate the enhancement of NO species both in atmosphere by emission spectrum and inside the solution by ultraviolet absorption spectroscopy. Moreover, qPCR analysis of oxidative stress mRNA shows higher gene expression. It is noted that 1% of HNO3 vapour plasma generates high amount of NO for killing bacteria and cancer cell killing.

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Enhancement of Nitric Oxide with nonthermal plasma jet and its effect on Escherichia coli inactivation

  • Shaw, Priyanka;Kumar, Naresh;Attri, Pankaj;Kwak, Hyong Sin;Choi, Eun Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.159-159
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    • 2015
  • A new approach for antimicrobial is based on the overproduction of reactive nitrogen species (RNS), especially; nitric oxide (NO) and peroxinitrite (ONOO-) are important factors to deactivate the bacteria. Recently, non-thermal atmospheric pressure plasma jet (APPJ) has been frequently used in the field of microbial sterilization through the generation of different kinds of RNS/ROS species. However, in previous study we showed APPJ has combine effects ROS/RNS on bacterial sterilization. It is not still clear whether this bacterial killing effect has been done through ROS or RNS. We need to further investigate separate effect of ROS and RNS on bacterial sterilization. Hence, in this work, we have enhanced NO production, especially; by applying a 1% of HNO3 vapour to the N2 based APPJ. In comparison with nitrogen plasma with inclusion of water vapour plasma, it has been shown that nitrogen plasma with inclusion of 1% of HNO3 vapour has higher efficiency in killing the E. coli through the high production of NO. We also investigate the enhancement of NO species both in atmosphere by emission spectrum and inside the solution by ultraviolet absorption spectroscopy. Moreover, qPCR analysis of oxidative stress mRNA shows higher gene expression. It is noted that 1% of HNO3 vapour plasma generates high amount of NO for killing bacteria.

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Effect of Iron(II)-ascorbate Complex on Protein and DNA of Phages (파아지 단백질 및 DNA에 대한 2가철-아스코르빈산착체의 영향)

  • Lho, Il-Hwan;Murata, Akira
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.46-51
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    • 1993
  • The inactivating effect of iron(II)-ascorbate complex (Fe-Asc) on various phages was previously reported. This paper describes the molecular target in the phage virion attacked by Fe-Asc. The effect of Fe-Asc on protein was investigated with bovine serum albumin and the structural protein of phage J1. There were no differences in the SDS-polyacrylamide gel electrophoresis (patterns of these two proteins when either they were treated) with Fe-Asc or not. Also, there were no changes in the amino acid composition and ultraviolet spectrum of the proteins. The effects of Fe-Asc on DNA was investigated with pUC18 DNA, M13mpB DNA and ${\lambda}$ DNA as well as DNA from phage J1. Fe-Asc caused initially nicking of the subsequently form of pUC18 DNA to yield the open circular form and then subsequently the linear form. Strand breaks were also confirmed with M13mp8 DNA and ${\lambda}$ DNA as well as J1 DNA. The results indicate that the strand breaks in phage DNA could be responsible for the inactivation of phages by Fe-Asc.

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Ribgrass Mosaic Tobamovirus Occurred on Chinese Cabbage in Korea

  • Kim, Jeong-Soo;Cho, Jeom-Deog;Choi, Hong-Soo;Lee, Soo-Heon;Choi, Gug-Seoun;Lee, Sang-Yong;Kim, Hye-Jeong;Yoon, Moo-Kyoung
    • The Plant Pathology Journal
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    • v.26 no.4
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    • pp.328-339
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    • 2010
  • A tobamovirus, Ribgrass mosaic virus (RMV), was identified newly from chinese cabbage (Brassica campestris L. pekinensis) in Korea. Virus disease incidence of RMV on chinese cabbage was 37.9% in alpine area on August in 1993. RMV induced the symptoms of necrotic ring spots, necrotic streak on midrib and malformation. RMV, Ca1 and Ca3 isolate, could infect 35 species out of 45 plants including Chenopodium amaranticolor. Physical properties of RMV Ca1 isolate were very stable as 10.8 over for dilution end point, $95^{\circ}C$ for temperature inactivation point and 18 weeks for longevity in vitro. RMV had the soil transmission rate of 75.0% for the chinese cabbages, 'Chunhawang' and 'Seoul' cultivars. The purified virions of RMV had the typical ultraviolet absorption spectrum of maximum at 260 nm and minimum at 247 nm. RMV of Ca1 isolate was related serologically with antisera of Tobacco mosaic virus (TMV)-Cym, TMV-O and Pepper mottle virus, but not related with antiserum of Odontoglossum ring spot virus. coat protein gene of RMV-Ca1, sized 473 nucleotides, encoded 158 amino acid residues. Nucleotide identity of RMV-Ca1 CP gene was 96.4% with RMV-Shanghai (GenBank accession No. of AF185272) from China and 96.0% with RMV-Impatiens (GenBank accession No. of AM040974) from Germany. Identity of amino acids between RMV-Ca1 and the two RMV isolates was 96.8%. Specific three primers were selected for rapid and easy genetic detection of RMV using Virion Captured (VC)/RT-PCR method.

Fabrication of a Water Sterilization System Utilizing a 275 nm-wavelength UVC LED and TIR Lens-equipped Light Source (275 nm UVC LED와 TIR 렌즈 장착 광원을 이용하는 물 살균장치 제작)

  • Kawan Anil;Seung Hui Yu;Seung Hoon Yu;J. A. Park;I. S. Shin;S. J. Lee;Y. B. Kim;Y. B. Kown;D. G. Han;Soon Jae Yu;Heetae Kim;Seong Bae Park
    • Journal of the Semiconductor & Display Technology
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    • v.23 no.1
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    • pp.84-87
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    • 2024
  • A water sterilization system is developed utilizing a 275 nm-wavelength LED light source equipped with a TIR lens. The system's light source is constructed by combining a 275 nm-wavelength UVC LED, known for its germicidal properties, with a TIR lens having a direction angle of 6.8 degrees. The optical simulation software 'LightTools' is employed to design and optimize the intensity of deep ultraviolet sterilizing light irradiation, its distribution, and sterilization capacity. In the inactivation experiment with E. coli, the water sterilizer system achieved a sterilization rate of 78.92 % while maintaining a water flow capacity of 50 L/min. Compared to the conventional mercury lamp light source water sterilizer system, the UVC LED water sterilizer system addresses environmental concerns related to mercury usage and offers advantages in terms of lifespan and durability.

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Studies on Mild Mutants of Tabacco Mosaic Virus I. Induction of Mild Mutants and Their Characteristics (약독 담배 모자이크 바이러스 I. 약독바이러스의 유기 및 특성)

  • Choi Jang Kyung;Son Kyung Ok
    • Korean Journal Plant Pathology
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    • v.1 no.1
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    • pp.3-11
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    • 1985
  • Three mild mutant strains of tobacco mosaic virus (TMV) were isolated from Nicotiana tabacum var. Samsun incubated at $38^{\circ}C$ for 10 days after inoculation with a wild type of TMV-OM strain. They were designated into Tg 5272, Tw 227 and Tw 333. All mild strains could be distinguished from TMV-OM by their reactions on different indicator plants. The mild strains induced the mild mottling without distinct symptoms, whereas the wild strain produced severe mosaic, rugose and stunting on tobacco and red pepper plants. Tw 227 and Tw 333 produced smaller necrotic spots than those of Tg 5272 and TMV-OM on N. glutinosa and Datura stramonium. The former two strains also produced ring spots and mosaic on Gomphrena globosa compared with necrotic spots by the latter strains. Three mild strains were serologically identical to TMV-OM. Their physical properties were thermal inactivation point $80-85^{\circ}C$, dilution end point between $10^{-4}\;and\;10^{-6}$, and longevity in vitro 7days or longer. Ultraviolet absorption spectra of purified preparations of the mild strains and TMV-OM were identical, with a minimum at 247nm, a maximum at 260nm, and a slight shoulder at 290nm. Electrophoresis of the strains in polyacrylamide-agarose gel showed that all the strains formed one major band and two minor bands, except for one minor band of Tw 333. However, when sodium dodecyl sulfate was added to the purified viruses before electrophoresis, each strain formed only one major band.

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Production of Induced Gynogenetic Diploid Bagrid Catfish Leiocassis ussuriensis (Siluriformes) - I. Optimization of Treatment Condition for Diploid Gynogenesis (대농갱이 Leiocassis ussuriensis (Siluriformes) 자성발생성 이배체 생산 - I. 자성발생성 이배체 유도 처리 조건의 최적화)

  • Park, Sang-Yong;Lee, Yoon-A;Nam, Yoon-Kwon;Bang, In-Chul
    • Journal of Aquaculture
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    • v.20 no.3
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    • pp.184-189
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    • 2007
  • Treatment conditions for the induced diploid gynogenesis, a maternal-exclusive form of artificial parthenogenetic reproduction, were optimized in bagrid catfish (Leiocassis ussuriensis, Siluriformes). Optimal amounts of ultraviolet (UV) irradiation for the genetic inactivation of spermatozoa in bagrid catfish and Pseudobagrus fulvidraco were proven to be ranged from 3,600 to 4,800 $ergs/mm^2$ based on the examination of viability of embryos and haploid incidence. Haploid embryos were restored to diploidy by preventing the extrusion of the second polar body using cold shock treatment. Thermal treatments (4 or $6^{\circ}C$ for 30, 40 or 50 min) were carried out 3, 5 or 7 min post insemination. Best scores for embryo viability (38.6% of total eggs taken) and incidence of normal diploidy (87.9% of hatched larvae) were observed at the embryo group treated at $4^{\circ}C$ for 40 min, 5 min after insemination. Restoration of gynogenetic diploidy was confirmed based on the absence of haploid syndrome, cell size and/or nucleolar organizing region (NOR) counts.

Studies on Cellulolytic Enzyme Producing by Ckaetomium globosum -Part 2. Purification of Cellulase- (Chaetomium globosum 이 생성하는 Cellulose 분해 효소에 관한 연구 -(제2보) Cellulase의 정제-)

  • Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.12
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    • pp.33-41
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    • 1969
  • 1. Crude cellulase extracted from wheat bran media of Chaetomium globosum with pH 7.0 McIlvaine buffer was fractionated by precipitation with ammonium sulfate and by treatment with the cellulose powder, DEAE-Sephadex A-25 and Amberite XE-65 (IRC-50) column chromatography. 2. Consquently two cellulases C-1 and C-2 were obtained by cellulose column chromatography. Cellulose C-1 was a powerful CMC-saccharifying and CMC-liquefying activity but cellulose C-2 was stronger CMC-liquefying activity compared to CMC-saccharifying activity and cellulase C-2 had smaller protein than that of cellulose C-1. And cellulose C-2 was fractionated by DEAE-Sephadex A-25 column chromatography into cellulase C-1-1 and cellulose C-1-2. 3. It can be obtained, therefore, that cellulose produced Chaelomium globosum consisted, at least, of three cellulases C-2, C-1-1 and C-1-2. 4. Cellulose C-1-1 was homogenous in the ultraviolet and the ultracentrifuge pattern. And cellulose C-1-1 had enzyme for CMC-saccharifying activity. 5. The optimum pH for the enzyme activity of cellulose C-1-1 was 4.0 in any methods of meas urement reducing sugar and viscosity. The optimum temperature was $40^{\circ}C$ in any methods. 6. The pH stability of cellulase C-1-1 was within pH 5.0 to pH 6.0 at $40^{\circ}C$ and fairly stable in acidic solution. 7. The heat stability was below $50^{\circ}C$ at pH 4.0 and complete heat inactivation of this cellulase occurred at $70^{\circ}C$.

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Photocatalytic disinfection of indoor suspended microorganisms (Escherichia coli and Bacillus subtilis spore) with ultraviolet light (광촉매와 UVA에 의한 실내 부유 미생물(E. coli 및 Bacillus. subtilis sp.) 살균 제거 연구)

  • Yoon, Young H.;Nam, Sook-Hyun;Joo, Jin-Chul;Ahn, Ho-Sang
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.2
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    • pp.1204-1210
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    • 2014
  • New control methods are proposed for indoor air quality by removing fine airborne dust-particles. As suspended fine dust-particles contain inorganic dust as well as fine organic bacteria, studies for simultaneous control of these contaminants are required. In this study, photocatalytic disinfection of indoor suspended microorganisms such as E. coli and Bacillus subtilis is performed by three types of photocatalysts with UVA irradiation. The UVA irradiation strength was controlled to the minimum $3{\mu}W/cm^2$, and ZnO, $TiO_2$, and ZnO/Laponite ball were used as the catalysts. The results indicate that E. coli was removed over 80 % after about 2 hours of reaction with UVA and all three types of photocatalysts, whereas only with UVA, around 50 % E. coli removal was obtained. Among the catalysts, ZnO/Laponite composite ball was found to have similar sterilizing capacity to $TiO_2$. However, in case of B. subtilis, which has thick cell wall in its spore state, disinfection was not effective under the low UVA irradiation condition, even with the catalysts. Further studies need to figure out the optimal UVA irradiation ranges as well as photocatalysts doses to control airborne dust, to provide healthy clean air environment.