• Title/Summary/Keyword: VP6

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Interaction of Heliothis armigera Nuclear Polyhedrosis Viral Capsid Protein with its Host Actin

  • Lu, Song-Ya;Qi, Yi-Peng;Ge, Guo-Qiong
    • BMB Reports
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    • v.35 no.6
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    • pp.562-567
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    • 2002
  • In order to find the cellular interaction factors of the Heliothis armigera nuclear polyhedrosis virus capsid protein VP39, a Heliothis armigera cell cDNA library was constructed. Then VP39 was used as bait. The host actin gene was isolated from the cDNA library with the yeast two-hybrid system. This demonstrated that VP39 could interact with its host actin in yeast. In order to corroborate this interaction in vivo, the vp39 gene was fused with the green fluorescent protein gene in plasmid pEGFP39. The fusion protein was expressed in the Hz-AM1 cells under the control of the Autographa californica multiple nucleopolyhedrovirus immediate early gene promoter. The host actin was labeled specifically by the red fluorescence substance, tetramethy rhodamine isothicyanete-phalloidin. Observation under a fluorescence microscopy showed that VP39, which was indicated by green fluorescence, began to appear in the cells 6 h after being transfected with pEGFP39. Red actin cables were also formed in the cytoplasm at the same time. Actin was aggregated in the nucleus 9 h after the transfection. The green and red fluorescence always appeared in the same location of the cells, which demonstrated that VP39 could combine with the host actin. Such a combination would result in the actin skeleton rearrangement.

Studies on the VP4 and VP7 Genes of Bovine Rotaviruses from Field Samples Using RT-PCR and RFLP Analysis (RT-PCR과 RFLP법을 이용한 국내 소 로타바이러스 VP4 및 VP7 유전자의 특성 규명)

  • Jeon, Seong-Jin;Chang, Chung-Ho;Chung, Chung-Won;Kim, Won-Yong;Kang, Shien-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.2
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    • pp.165-174
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    • 1998
  • Characterizations of the VP4 (P type) and VP7 (G type) genes of Korean isolates of bovine rotavirus were performed using RT-PCR/RFLP and nucleotide sequencing analysis. After RT-PCR amplification of partial length (1094bp) of the VP4 and full length (1062bp) of the VP7 genes, amplified PCR products were digested with restriction endonucleases and digestion patterns were compared with those of reference rotaviruses. With the VP4 genes, four RFLP (A-D) profiles were observed; three (A, Band C) were the same as those of bovine rotavirus NCDV (P[1]), IND (P[5]) and B223 (P[11]), respectively. Profile D was the same as that of porcine rotavirus OSU (P[7]). With the VP7 genes, five RFLP profiles (I-V) were observed; three of them (I, II and III) were the same as those of bovine rotavirus NCDV (G6), Cody 1-801 (G8), and B223 (G10), respectively. Profile IV and V were atypical to those of reference bovine rotaviruses used in this study. These two profiles were identified as G6 and G5, respectively, after analyzing and comparing the nucleotide sequences. The G typing analysis revealed that 61.9% (26/42) were G6, which included G6 subtype; 28.6% (12/42) were G5; 7.1% (3/42) were G10; 2.4% (1/42) were G8. The P typing analysis revealed that 54.8% (23/42) were P[5]; 28.6% (12/42) were P[7]; 11.8% (5/42) were [11]; 4.8% (2/42) were P[1]. Our results showed that G6/P[5] were the most prevalent rotaviruses in diarrheic calves in Korea. Also, this is the first report that G5/P[7] rotaviruses were identified from cattle with diarrhea.

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Optimal Exposure Conditions according to Detector Type in Chest Digital Radiography (디지털흉부X선촬영에서 검출기 방식에 따른 최적의 노출조건)

  • Lee, Won-Jeong
    • Journal of the Korea Safety Management & Science
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    • v.17 no.4
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    • pp.213-221
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    • 2015
  • The aim of this study was to set up the optimal exposure condition according to detector type considering image quality (IQ) with radiation dose in chest digital radiography. We used three detector type such as flat-panel detector (FP) and computed radiography (CR), and charge-coupled device (CCD). Entrance surface dose (ESD) was measured at each exposure condition combined tube voltage with tube current using dosimeter, after attaching on human phantom, it was repeated 3 times. Phantom images were evaluated independently by three chest radiologists after blinding image informations. Standard exposure condition using each institution was 117 kVp-AEC at FP and 117 kVp-8 mAs at CR, and 117 kVp-8 mAs at CCD. Statistical analysis was performed by One way ANOVA (Dunnett T3 test) using SPSS ver. 19.0. In FP, IQ scores were not significant difference between 102 kVp-4 mAs and 117 kVp-AEC (28.4 vs. 31.1, p=1.000), even though ESD was decreased up to 50% ($62.3{\mu}Gy$ vs. $125.1{\mu}Gy$). In CR, ESD was greatly decreased from 117 kVp-8 mAs to 90 kVp-8 mAs without significant difference of IQ score (p=1.000, 24.6 vs. 19.5). In CCD, IQ score of 117 kVp-8 mAs was similar with 109 kVp-8 mAs (29.6 vs. 29.0), with decreasing from $320.8{\mu}Gy$ to $284.7{\mu}Gy$ (about 11%). We conclude that optimal x-ray exposure condition for chest digital radiography is 102 kVp-4 mAs in FP and 90 kVp-8 mAs in CR, and 109 kVp-8 mAs in CCD.

Efficacy of Genetic Adjuvant (Plasmid-Expressed Chicken Interleukin-6) and Chemical Adjuvant (Levamisole) on the Protective Immunity of Genetic Vaccine against Infectious Bursal Disease Virus (닭의 전염성 F낭병 바이러스 유전자백신에 의한 방어 면역에 Genetic Adjuvant (Chicken Interleukin-6)와 Chemical Adjuvant (Levamisole)의 효과)

  • Park, Jeong-Ho;Sung, Haan-Woo;Yoon, Byung-Il;Pak, Son-Il;Kwon, Hyuk-Moo
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.91-98
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    • 2009
  • Infectious bursal disease (IBD) caused by the infectious bursal disease virus (IBDV) has an important economic impact on the poultry industry worldwide. This study examined the adjuvant effects of a plasmid encoding chicken interleukin-6 (pcDNA-ChIL-6) and levamisole (LMS) on in ovo prime-boost vaccination using a genetic vaccine (pcDNA-VP243) to prime in chicken followed by a killed-vaccine boost. A pcDNA-VP243 was injected into the amniotic sac alone or in combination with a pcDNA-ChIL-6 or LMS at embryonation day 18, followed by an intramuscular injection of killed IBD vaccine at 1 week of age. The chicken were orally challenged with very virulent IBDV (vvIBDV) strain at 3 weeks of age and observed for 10 days. No mortality was observed in the groups that received the pcDNA-VP243 alone and pcDNA-VP243 plus pcDNA-ChIL-6 or LMS compared to 100% mortality in unvaccinated challenge control group. However, as determined by bursal damage (the presence of IBDV RNA, B/B ratio, and lesion score), a pcDNA-VP243 alone group was superior to pcDNA-VP243 plus pcDNA-ChIL-6 or LMS groups in the protection against post-challenge. These findings suggest that in ovo priming with genetic vaccine and boosting with killed vaccine is an effective strategy for protecting chicken against vvIBDV and the addition of pcDNA-ChIL-6 or LMS did not enhance protective immunity.

Monoclonal antibodies against porcine group C rotavirus VP6 (돼지 group C 로타바이러스 VP6 특이 단클론항체)

  • Yoon, Young-Sim;Lee, Seung-Chul;Woo, Sang-Kyu;Cho, Kyoung-Oh;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.3
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    • pp.175-182
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    • 2012
  • Rotaviruses have been known to be a major etiological agent of gastroenteritis in both infants and young animals. Subsequently new rotaviruses, which were morphologically indistinguishable but antigenically and electrophoretically distinct with each other, were reported from several animals throughout world including Korea. These new rotaviruses were named as non-group A or group B or group C rotaviruses and so on. It has been very difficult to isolate and grow the non-group A rotaviruses in vitro, and this has greatly limited the characterizations of non-group A rotaviruses and serological studies. In this study, monoclonal antibodies (MAbs) against porcine non-group A rotavirus were produced and characterized. The VP6 gene of porcine group C rotavirus Korean isolate(#06-52-1) was cloned and expressed. For expression of VP6 gene, baculovirus expression system was applied. The VP6 gene and expressed protein in the recombinant virus were confirmed by polymerase chain reaction (PCR), indirect fluorescence antibody (IFA) test and Western blot, respectively. The expressed VP6 was used for MAbs production. The MAbs produced in this study would be promising as diagnostic reagents for detection of group C rotavirus infection.

Evaluation of Image Quality and dose with the Change of kVp and BMI in the Liver CT (CT 검사 시 관전압과 BMI 변화에 따른 화질 및 피폭평가)

  • Kim, Dong-Hyun;Ko, Sung-Jin;Kang, Se-Sik;Kim, Jung-Hoon;Choi, Seok-Yoon;Kim, Changsoo
    • The Journal of the Korea Contents Association
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    • v.13 no.6
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    • pp.331-338
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    • 2013
  • CT for follow-up visits because of liver disease, body mass index (BMI) and kVp according to the change of the image quality and radiation dose to evaluate for changes. March 2010 to June 2011 at Pusan P University Hospital, abdominal CT scans a patient BMI (Body Mass Index. Less BMI) index was less than 25 in the treatment of subjects had a 48-person Noise and SNR at 100kVp abdominal image is lager than the 120kVp image. CTDI volume value at by the analysis of the radiation dose is 4.47mGy(100kVp) and 9.01mGy(120kVp). So CTDIvol in 100kVp is smaller than CTDIvol in 120kVp(decrease by 44.1%). And, effective dose is 7.1mSv(100kVp) and 12.51mSv(120kVp). So effective dose in 100kVp is smaller than effective dose in 120kVp(decrease by 43%). Evaluation of image quality is that Unacceptable 0 person, Suboptimal 0 person, Adequate 0 person, Good 1 person, Excellent 47 person. In case of repeatly patient, we examinate abdomianl CT scan by using low kVp and body mass index less than 25. We can has good quality image and benefit of low radiation dose.

Design of Unified Inverse Transformer for HEVC and VP9 (HEVC 및 VP9 겸용 통합 역변환기의 설계)

  • Jung, Seulkee;Lee, Seongsoo
    • Journal of IKEEE
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    • v.19 no.4
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    • pp.596-602
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    • 2015
  • In this paper, a unified inverse transformer is designed for HEVC and VP9. The proposed architecture performs all modes of HEVC and VP9 in the unified inverser transformer, such as $4{\times}4{\sim}32{\times}32$ HEVC IDCT, $4{\times}4$ HEVC IDST, $4{\times}4{\sim}32{\times}32$ VP9 IDCT, $4{\times}4{\sim}16{\times}16$ VP9 IADST and $4{\times}4$ IWHT. Same computations are used in HEVC IDCT and VP9 IDCT, except for the scales of the coefficients. Similarly, same computations are used in HEVC $4{\times}4$ IDST and VP9 $4{\times}4$ IADST, except for the scales of the coefficients. Furthermore, HEVC IDCT, VP9 IDCT, and VP9 IADST are the subsets of upper level IDCTs. The proposed architecture reuses multipliers when the computation is identical. Also it shares adders and butterfly structures even when the multiplier coefficients are different. So it reduces the hardware size significantly. Synthesized in 0.18 um technology, the gate count is 456,442 gates. which achieved 22.6% reduction compared to conventional architectures.

Usefulness of Escherichia coli-expressed Recombinant VP6 Proteins of Group A Rotavirus in Serodiagosis of Rotavirus Infection (대장균에서 발현된 A군 로타바이러스 VP6 단백질을 이용한 로타바이러스 감염의 혈청학적 진단의 유용성)

  • Seo, Ji-Hyun;Kim, So-Young;Park, Ji-Sook;Lim, Jae-Young;Park, Chan-Hoo;Woo, Hyang-Ok;Youn, Hee-Shang;Kim, Won-Yong;Kang, Hyung-Lyun;Baik, Seung-Chul;Lee, Woo-Kon;Cho, Myung-Je;Rhee, Kwang-Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.13 no.2
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    • pp.134-145
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    • 2010
  • Purpose: The serologic diagnosis of rotaviral infections is not commonly used in clinical practice, but is used in seroepidemiologic studies. In this study, the usefulness of Escherichia coli-expressed recombinant VP6 proteins of group A rotavirus in the serodiagnosis of rotavirus infections by ELISA was evaluated. Methods: The recombinant VP6 proteins of group A rotavirus expressed in E. coli Rosetta II strain were purified and identified. One hundred sera from 22 children (4 healthy neonates, 13 healthy children, and 5 immunocompromised children) who had serial sera samples prior to and after rotavirus infections were provided by the Gyeongsang National University Hospital, a member of the National Biobank of Korea. IgG, IgA, and IgM antibodies against rVP6 were analyzed by ELISA in all of the patients and Western blot analysis in 4 neonates. Results: ELISA tests using rVP6 proteins of group A rotavirus as antigen revealed that IgG, IgA, and IgM antibodies increased after rotaviral infections in most neonates and healthy children. IgG antibodies also increased after rotaviral infections in most immunocompromised children without an adequate increase in IgM or IgA antibodies. Western blot analysis in four neonates revealed very early IgM antibody responses, even in the sera with low optical densities in ELISA tests. Conclusion: Our study showed that ELISA using rVP6 as an antigen is a valid diagnostic tool for seroepidemiologic studies of rotavirus infections and Western blot analysis is a sensitive test in detecting IgG, IgA, and and IgM antibodies in patients with rotavirus infections.

Response of Electrocardiogram of Nile tilapia, Oreochromis niloticus to Electric Stimulus (전기자재에 대한 역돔의 심전도)

  • 한규환;양용림
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.38 no.4
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    • pp.278-283
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    • 2002
  • The response of electrocardiogram(ECG) of Nile tilapia, Oreochromis niloticus [Linnaeus] was studied to the electric stimulus which was given to a certain part of body The experiments were performed in such a way that three levels of electric stimulus (20, 30, 40 Vp ; 10 msec) were given to fishes with electrode inserted into their bodies and then their ECGs were recorded continuously for 60 minutes in the water temperature of 16~18$^{\circ}C$ The results of the experiments were divided by day and night, and then were analyzed by experimental conditions as follows; 1. Nile tilapia reached a stable condition within 3 minutes after the electrode inserted into their bodies during anesthesia. In stable condition, the heart rates average was 45.8 beat/min during daytime and 45.0 beat/min at night. The action potentials average was 1.76 $mutextrm{V}$during daytime and 1.75 $mutextrm{V}$ at night. 2. The heart rates average by three levels of electric stimulus were \circled1 In the stimulus condition, the heart rates were 34.9 beat/min during daytime and 33.4 beat/min at night for the 20 Vp level, 36.8 bea/min during daytime and 36.0 beat/min at night for the 30 Vp level, and 38.0 beat/min during daytime and 36.4 beat/min at night for the 40Vp level. \circled2 In the recovery condition, the action potentials were 45.5 beat/min during daytime an 45.1 beat/min at night for the 20Vp level, 47.9 beat/min during daytime and 49.0 beat/min at night for the 30Vp level, and 51.4 beat/min during daytime and 50.7 beat/min at night for the 40Vp level 3. The action potentials average by three levels of electric stimulus were, \circled1 In the stimulus condition, action potentials were 2.54 $mutextrm{V}$ during daytime and 2.39 $mutextrm{V}$ at night for the 20 Vp level, 3.30 $mutextrm{V}$ during daytime and 2.30 $mutextrm{V}$ at night for the 30 Vp level and 6.05 $mutextrm{V}$ during daytime and 3.23 $mutextrm{V}$ at night for the 40 Vp level. \circled2 In the recovery condition, action potentials were 1.92 $mutextrm{V}$ during daytime and 1.95 $mutextrm{V}$ at night for the 20 Vp level and 2.78 $mutextrm{V}$ during daytime and 2.21 $mutextrm{V}$ at night for the 30Vp level and 3.6 0 $mutextrm{V}$ during daytime and 2.98 $mutextrm{V}$ at night for the 40 Vp level.

Use of the Synthetic Gene Encoding the Truncated Human Rotavirus VP8* Protein in Escherichia coli for Production of Vaccine Candidates or Development of Diagnostic Antibodies (합성 유전자를 이용하여 Escherichia coli에서 백신 후보의 생산 혹은 진단용 항체의 개발을 위한 인간 rotavirus VP8* 부분 단백질의 발현)

  • Kim, Sang-Rae;Lee, Bheong-Uk
    • Journal of Life Science
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    • v.28 no.4
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    • pp.478-482
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    • 2018
  • Human rotavirus is a causative agent of acute diarrhea among children. The artificial gene encoding the truncated $VP8^*$ protein of human rotavirus A (serotype 1 strain WA) was synthesized according to the Escherichia coli codon preference. The synthetic $VP8^*$ gene also possessed the NdeI and HindIII restriction sites for the convenient in-frame cloning for translation and a 6-histidine tag at C-terminus for Ni+ affinity purification. Molecular weight of the truncated $VP8^*$ protein deduced from the nucleotide sequences of the artificial gene was a 19.7-kDa. This synthetic $VP8^*$ DNA fragment was inserted into the pT7-7 expression vector and transformed into E. coli BL21 (DE3). Transformants harboring the synthetic gene encoding the $VP8^*$ protein was induced by supplement of a final concentration of 0.05 mM ITPG at $20^{\circ}C$. Protein crude extract from the E. coli transformants was subjected to Western blotting with the mouse anti-rotavirus capsid antibody, showing ~20-kDa $VP8^*$ protein band. The truncated $VP8^*$ protein band was also observed by Western blotting using the rabbit polyclonal antibody serum made against the truncated $VP8^*$ protein. This study suggested that the synthetic gene could be used as an easy way to produce the antigenic vaccine candidate for control of virus-associated diseases or to develop antibodies for diagnostic purpose.