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Characterization of gp64 Gene of Bombyx mori Nucleopolyhedrovirus and Development of a Transient Expression Vector (누에 핵다각체병 바이러스 헤 gp64 유전자의 특성조사 및 transient 발현 벡터 개발)

  • 김미향;최재영;우수동;이해광;제연호
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.18-24
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    • 2001
  • Expression of the baculovirus major envelope glycoprotein gene(gp64) is regulated by transcription from botha early and late promoters. To develop a transient expression vector under the control of gp64 gene promoter, the gp64 gene of Bombyx mori nucleopolyhedrovirus-K1(BmNPV-K1) was characterized. The gp64 gene was local-ized at EcoR I-Pst I 7.38-kb fragment of the BmNPV-K1 genome. The EcorR 1-Pst I 7.38-kb fragment was cloned and the nucleotide sequence of 2,277 bases including the coding region of gp64 gene was determined. Based on these results, transient expression vector using gp64 gene promoter was constructed and named as pBm64. E.coli lacZ gene was introduced onto pBm64 as a reporter gene and expressed transiently in B. mori 5(Bm 5) cells. The expression vector transfected into the cells was maintained stably for 1 to 5 days. In order to confirm the expression of the reporter gene by gp64 promoter, recombinant virus was constructed. The recombinant virus has two independent transcription units in opposite orientations with two promoters; gp64 and polyhedrin gene promoters each initiating transcription of $\beta$-galactosidase and polyhedrin, respectively. Polyhedra formation and expression of $\beta$-galactosidase in Bm5 cells infected with the recombinant virus were observed with phase contrast microscope and in situ staining.

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Shear behavior at the interface between particle and non-crushing surface by using PFC (PFC를 이용한 입자와 비파쇄 평면과의 접촉면에서의 전단 거동)

  • Kim, Eun-Kyung;Lee, Jeong-Hark;Lee, Seok-Won
    • Journal of Korean Tunnelling and Underground Space Association
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    • v.14 no.4
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    • pp.293-308
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    • 2012
  • The shear behavior at the particle/surface interface such as rock joint can determine the mechanical behavior of whole structure. Therefore, a fundamental understanding of the mechanisms governing its behavior and accurately estimation of the interface strength is essential. In this paper, PFC, a numerical analysis program of discrete element method was used to investigate the effects of the surface roughness on interface strength. The surface roughness was characterized by smooth, intermediate, and rough surface, respectively. In order to investigate the effects of particle shape and crushing on particle/surface interface behavior, one ball, clump, and cluster models were created and their results were compared. The shape of particle was characterized by circle, triangle, square, and rectangle, respectively. The results showed that as the surface roughness increases, interface strength and friction angle increase and the void ratio increases. The one ball model with smooth surface shows lower interface strength and friction angle than the clump model with irregular surface. In addition, a cluster model has less interface strength and friction angle than the clump model. The failure envelope of the cluster model shows non-linear characteristic. From these findings, it is verified that the surface roughness and particle shape effect on the particle/surface interface shear behavior.

Discrimination of Flaviviruses with High Frequency of Infection in Asian Countries: Epitope Prediction by Bioinformatic Approaches (아시아 국가 내에서 감염빈도가 높은 플라비바이러스의 구별: 생물정보학적 접근을 통한 항원결정기 예측)

  • Choi, Jae-Won;Jo, Byung-Gwan;Kim, Min Jung;Park, Suji;Kim, Hak Yong
    • The Journal of the Korea Contents Association
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    • v.18 no.4
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    • pp.99-113
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    • 2018
  • Recently, global warming has widened the habitat of mosquitoes and infection chances for mosquito-borne diseases are increasing. Flavivirus is a typical mosquito-borne virus. Flaviviruses with a relatively high frequency of infection in Asian countries include Zika, Dengue, and Japanese encephalitis viruses. Although distinctive diagnosis of flaviviruses is required because the symptoms and therapeutic method differ, there is no diagnostic method that can distinguish them accurately yet. In this study, we propose distinctive diagnosis method of flaviviruses using informations and analysis tools constructed in bioinformatic databases. The envelope protein and non-structural protein 1 which are useful protein for the immuno-diagnostics of three flaviviruses were selected. Their homology was analyzed by multiple sequence alignments and epitope candidates consisting of 10-15 amino acids were selected. Finally two epitopes were suggested to be most useful by immunogenicity analysis and 3D structure prediction. These approaches and results are expected to be great value in the distinctive diagnosis of three flaviviruses with a high frequency of infection in Asian countries.

Predicting Parturition Time through Ultrasonic Measurement of Posture Changing Rate in Crated Landrace Sows

  • Wang, J.S.;Wu, M.C.;Chang, H.L.;Young, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.682-692
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    • 2007
  • This study presents an automatic system to predict parturition time in the crated sows. The system relies on ultrasonic transducers mounted from above along the length of the crate. Using a 40 kHz time of flight (TOF) single envelope wave, the momentary distances between the sensors are measured. Therefore, the local momentary height of the sow and the momentary posture, i.e. standing posture (SDP), kneeling posture (KP), sitting posture (STP) and lateral lying posture (LLP) are determined. Crated sows change their postures from standing to lying and vice versa which follows a characteristic pattern. As parturition approaches, sows exhibit uneasiness, restlessness and the stand up sequence (SUS, the posture transition from LLP to SDP) rate increases because of labor pains. In time series, the SUS rate demonstrates a peak and it happens approximately 0-12 h before parturition. In this paper, the basic parturition threshold value method (BPTVM) and the same hour method (SHM) are proposed for predicting parturition, both of which are based on the SUS rate. The BPTVM mainly detects the peak of the SUS rate. As the SUS rate exceeds the threshold value, the parturition becomes predictable. Moreover, the SHM calculates the difference in the SUS rates between a particular time of day and the corresponding time of the preceding day. Compared to the BPTVM, the SHM can eliminate the circadian rhythm of the SUS rate influenced by feeding behavior. Using the SHM the parturition can be approximately predicted within hours. In an attempt to define the threshold parameters of predicting parturition, a data set with 32 sows of the SUS rate are used to estimate assumable predicting probability. The results show the assumable probability of the parturition prediction within 9 h is 96.9% for the SHM and 84.4% for the BPTVM. Moreover, the SHM can even reach a 75% probability of prediction within three hours of parturition. We conclude that the SHM is more accurate and is more useful for parturition time prediction. When parturition is detected, the proposed algorithm generates a warning signal which can inform human personnel to protect the mother and newborn piglets.

Filamentous growth of Escherichia coli by dephosphorylated NPr (탈인산화된 NPr에 의한 대장균의 섬유상 생장)

  • Choi, Umji;Seok, Yeong-Jae;Lee, Chang-Ro
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.149-155
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    • 2017
  • The nitrogen phosphotransferase (PTS) system is a regulatory cascade present in most Proteobacteria, where it controls different functions. The nitrogen PTS is usually composed of $EI^{Ntr}$ (encoded by the ptsP gene), NPr (encoded by the ptsO gene), and $EIIA^{Ntr}$ (encoded by the ptsN gene). While $EIIA^{Ntr}$ plays a role in a variety of cellular processes, such as potassium homeostasis, regulation of ppGpp accumulation, nitrogen and carbon metabolisms, and regulation of ABC transporters, little information is available for a physiological role of NPr. A recent study showed that dephosphorylated NPr affects adaptation to envelope stresses in Escherichia coli. In this study, we provide another phenotype related to NPr. The ptsP mutant showed a filamentation phenotype. The filamentation phenotype of the ptsP mutant was recovered by additional deletion of the ptsO gene, but not by additional deletion of the ptsN gene, suggesting that an increased level of dephosphorylated NPr in the ptsP mutant renders cells the filamentous growth. This idea was confirmed by the fact that cells with increased levels of dephosphorylated NPr shows the filamentation phenotype. Additionally, we showed that cell size of E. coli increases with incremental dephosphorylated NPr concentrations. These results suggested that dephosphorylated NPr induces morphological change of E. coli.

Vibration Analysis of SAR Antenna Reflectors During Satellite Maneuver (위성 기동 시 SAR 안테나 반사판에 발생하는 진동 분석)

  • Kim, Tae-Hyun;Kim, Dae-Yeon;Suh, Jong-Eun;Han, Jae-Hung;Lee, Jae-Eun;Jung, Hwa-Young
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.48 no.3
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    • pp.225-231
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    • 2020
  • Recently, there has been an increasing demand for SAR satellite as it can be operated regardless of the weather condition. In general, main reflector of the SAR is formed of multiple deployable panels to increase performance in the constrained payload envelope. By nature, deployable structure lacks structural stiffness and it is vulnerable to external disturbances and excitation. In particular, SAR satellites may have high levels of vibration occurring at the antenna reflecting surface due to higher angular rate requirements. During image capturing it is important to keep high surface accuracy of the reflector for the quality of images. In this research, a performance degradation of deployable SAR antenna due to structural deformation is analyzed. Panels for main reflectors are assumed to be flexible structures and multi-body simulation environment is established. Then, deflection of the panel is calculated while the satellite performs maneuvers. In addition, antenna gain and beam pointing error are analyzed to determine how these deflections affect antenna performance and mission.

Experimental Study on the Shear Strength Characteristics of the Saturated Sand (포화(飽和)모래의 전단강도특성(剪斷强度特性)에 관(關)한 실험적(實驗的) 연구(研究) -대구지역(大邱地域) 낙동강(洛東江) 모래에 대해-)

  • Kim, Young Su;Seo, In Shik;Kim, Byoung Tak
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.14 no.6
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    • pp.1417-1431
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    • 1994
  • In this study, shear strength characteristics of the Nak-Dong river sand in Taegu area are investigated by triaxial compression test, considering shear strain control velocity, relative density, and confining pressure. The results from Lade model and Modified Lade model are compared with the measured value in the laboratory test. The results by the change of shear strain control velocity, relative density, and confining pressure are as follows; 1) The stress limit, which can be Coulomb's law about the Nak-Dong river sand, have ranged 120~200 kpa at 0.08%/min and 120~150 kpa at 0.5%/min. These limits are smaller than that of the calcareous sand and the well-graded, quartz sand. 2) The parameters needed to Lade model and Modified Lade model are much affected by the strain control velocity and the relative density. Consequently, in the field, it is important to use parameters aptly after accurately understanding both the loading condition and subsoil condition. 3) Overall, the principal stress ratio obtained from constitutive model equations is not affected by the control velocity, but both the relative density and confining pressure affect the result of constitutive model equations. Consequently, the study on the various conditions about the relative density and confining pressure is needed to accurately predict the stress-strain behavior on the Nak-Dong river sand. 4) For the range of the used confining pressure in the study, the Lade model shows better agreements with the measured value than the Modified Lade model, comparing the measured value with the principal stress ratio at failure and the internal friction angle of failure envelope obtained from the Lade model and Modified Lade model.

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Ultrastructural Description on Oogenesis of the Melania Snail, Semisulcospira libertina libertina (Gastropoda: Pleuroceridae) (다슬기, Semisulcospira libertina libertina의 난자형성과정에 관한 미세구조적 기재)

  • Kim, Eun-Kyoung;Lee, Jung-Sick
    • The Korean Journal of Malacology
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    • v.25 no.2
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    • pp.145-151
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    • 2009
  • The ultrastructural changes in germ cells during oogenesis of the melania snail, Semisulcospira libertina libertina have been investigated by light and electron microscopy. The ovary is located on the surface of the hepatopancreas in the spiral posterior region. The ovary exhibited greenish color in the gonadal mature season. The ovary was composed of a number of oogenic follicles. Oogenesis was divided into five stages with histological features: (1) oogonia, (2) previtellogenic, (3) initial vitellogenic, (4) active vitellogenic, and (5) mature stages. Oogonia were oval in shape, $4-6\;{\mu}m$ in diameter, and had a large nucleus. Previtellogenic oocytes were about $20\;{\mu}m$ in diameter and the cytoplasm reacted with hematoxylin in H-E satin. Initial vitellogenic stage, oocytes were $60-80\;{\mu}m$ in diameter, and small yolk granules of low electron density are scattered in the cytoplasm. Oocytes in the initial vitellogenic stage were connected with ovarian follicle by egg stalk. Active vitellogenic oocyte were $100-120\;{\mu}m$ in diameter. Electron density, size and quantity of yolk granules that are distributed in the cytoplasm have increased from the previous stage. Result of TEM observations, the oocyte contains well-developed Golgi complex, endoplasmic reticula and tubular mitochondria in the cytoplasm. Cytoplasm of mature oocyte was filled with proteinaceous yolk globules of high electron density. In this stage, the length of microvilli in the egg envelope was approximately $1.1\;{\mu}m$.

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Hepatitis C Virus - Proteins, Diagnosis, Treatment and New Approaches for Vaccine Development

  • Keyvani, Hossein;Fazlalipour, Mehdi;Monavari, Seyed Hamid Reza;Mollaie, Hamid Reza
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.5917-5935
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    • 2012
  • Background: Hepatitis C virus (HCV) causes acute and chronic human hepatitis infection and as such is an important global health problem. The virus was discovered in the USA in 1989 and it is now known that three to four million people are infected every year, WHO estimating that 3 percent of the 7 billion people worldwide being chronically infected. Humans are the natural hosts of HCV and this virus can eventually lead to permanent liver damage and carcinoma. HCV is a member of the Flaviviridae family and Hepacivirus genus. The diameter of the virus is about 50-60 nm and the virion contains a single-stranded positive RNA approximately 10,000 nucleotides in length and consisting of one ORF which is encapsulated by an external lipid envelope and icosahedral capsid. HCV is a heterogeneous virus, classified into 6 genotypes and more than 50 subtypes. Because of the genome variability, nucleotide sequences of genotypes differ by approximately 31-34%, and by 20-23% among subtypes. Quasi-species of mixed virus populations provide a survival advantage for the virus to create multiple variant genomes and a high rate of generation of variants to allow rapid selection of mutants for new environmental conditions. Direct contact with infected blood and blood products, sexual relationships and availability of injectable drugs have had remarkable effects on HCV epidemiology. Hundreds of thousands of people die each year from hepatitis and liver cancer caused by HCV virus infection. Approximately 80% of patients with acute hepatitis C progress into a chronic disease state leading to serious hepatic disorders, 10-20% of which develop chronic liver cirrhosis and hepatocellular carcinoma. The incubation period of HCV is 6-8 weeks and the infection is often asymptomatic so it is very hard to detect at early stages, making early treatment very difficult. Therefore, hepatitis C is called a "silent disease". Neutralizing antibodies are produced against several HCV proteins during infection but the virus mutates to escape from antibodies. Some patients with chronic hepatitis C may have some symptoms such as fatigue, muscle aches, nausea and pain. Autoimmune and immunecomplex-mediated diseases have also been reported with chronic HCV infection.

Light and Electron Microscopy of Rainbow Trout Gonadotropes and Spermatogenic Cells (무지개송어 뇌하수체의 성선자극호르몬 분비세포와 정자형성세포의 광학 및 전자현미경적 미세구조)

  • Yoon, Jong-Man;Park, Hong-Yang
    • Development and Reproduction
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    • v.2 no.1
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    • pp.89-99
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    • 1998
  • This study was carried out to investigate the morphological changes of gonadotropes in pituitary gland and spermatogenic cells in testis, obtained from 150 of 3-year-old immature and mature male rainbow trout (Oncorhynchus mykiss) during the reproductive cycles from March to February in the following year. In the maturation cycle of the pituitary gonadotropes of cultured rainbow trout, three periods can be distinguished i.e. a period of resting(March-August), a period of full spermatogenesis (September-November), and a period of breeding (December-February). The ultrastructures of the gonadotropes largely parallel the cyclical changes in the tests. The seminiferous tubules contain all spermatogenetic stages and sperm cells in a period of early maturation. At first, the size of the nucleus and cytoplasm decrease gradually at every stages from spermatogonia to spermatids. In the secondary spermatocytes, the small mitochondria are located over the outer cytoplam. In spermatids, the cytoplasmic masses move toward the posterior part of the nucleus. In spermatids, the two large mitochondria are located over the cytoplasm. In spermatids, the cytoplasmic masses move towark the posterior part of the nucleus. In spermatids, the two large mitochondria are located over the cytoplasm and begin to elongate. In spermatozoa, the surface of the nucleus devreases in volume. Examination by TEM shows that the nuclear envelope and plasma membrane are slightlywrinkled and closely adhered to the nucleus of spermatozoa. Two oval mitochondria are quite separated and the flagellum is inserted into the base of the spermatozoa head.The axoneme in this fish has the typical pattern such as nine peripheral doublets and a central doublet(9+2). there are remarkable individual differences in the size and morphology of spermatozoa head as observed by transmission and scanning electron microscopy.

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