• Title/Summary/Keyword: inclusion bodies

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Occurrence of Hydropericardium-Hepatitis Syndrome of Baeksemi and Broiler Raised in Korea

  • Seon, Jeong-Won;Ha, Jong-Soo;Do, Seon-Hee;Kim, Ki-Seuk
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2005.11a
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    • pp.80-81
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    • 2005
  • The occurrence of HHS was confirmed for the first time in Korea from chickens submitted for diagnosis to our laboratory from broiler and baeksemi farms. Clinical signs included depression, inappetence, ruffled feathers and a increase in mortality. At necropsy, severe hydropericardium and hepatic necrosis was founded characteristically and the most remarkable microscopic changes were seen in the liver. These included basophilic intranuclear inclusion bodies in the hepatocytes, massive hemorrhages and necrosis in the liver parenchyma. We could also identify fowl adeno-virus(FAV) by polymerase chain reaction(PCR) and electro-microscopic confirmation. Abbreviation: HHS=hydropericardium hepatitis syndrome, EM=electron microscopy, FAV=fowl adenovirus, PCR=polymerase chain reaction.

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Thoracic teratoma in a laying hen (Deklb brown warren). A case report (산란계에서의 흉강내 기형종 발생보고)

  • 한규삼;이성효;서문정;오언평;채효석;유혜진;임채웅
    • Korean Journal of Veterinary Service
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    • v.20 no.2
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    • pp.183-189
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    • 1997
  • This article reports a spontaneous thoracic teratoma in a laying hen (Deklb brown warren). In this case, a 40 days old hen was submitted for necropsy as part of an investigation into a flock problem suspected Marek's disease to Chonbuk Veterinary Service Laboratory in 1983. On the gross finding, mass was In the cranial subpulmonary cavity and attached to the vertebral column. It contained fully developed contour feathers. Histologically, feathers were arised from feather follicles complete with arrector plumi muscle, nerve, vessel and mucous gland composed with simple tall columnar epithelium. The outer surface of mass was lined by keratinized or nonkeratinized stratified squamous epithelium. It converted to pseudostratified ciliated columnar epithelium in some area. There were lymphocyte infiltration around gland tissue and eosinophilic intranuclear inclusion bodies in nonkeratinized epithelium. This thoracic teratoma was composed of ectodermal origin, squamous epithelium and nerve, and endodermal origin, mucous gland. This case in laying hen has never been reported in the literature in the world.

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Patient-specific pluripotent stem cell-based Parkinson's disease models showing endogenous alpha-synuclein aggregation

  • Oh, Yohan
    • BMB Reports
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    • v.52 no.6
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    • pp.349-359
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    • 2019
  • After the first research declaring the generation of human induced pluripotent stem cells (hiPSCs) in 2007, several attempts have been made to model neurodegenerative disease in vitro during the past decade. Parkinson's disease (PD) is the second most common neurodegenerative disorder, which is mainly characterized by motor dysfunction. The formation of unique and filamentous inclusion bodies called Lewy bodies (LBs) is the hallmark of both PD and dementia with LBs. The key pathology in PD is generally considered to be the alpha-synuclein (${\alpha}$-syn) accumulation, although it is still controversial whether this protein aggregation is a cause or consequence of neurodegeneration. In the present work, the recently published researches which recapitulated the ${\alpha}$-syn aggregation phenomena in sporadic and familial PD hiPSC models were reviewed. Furthermore, the advantages and potentials of using patient-derived PD hiPSC with focus on ${\alpha}$-syn aggregation have been discussed.

Enhanced Enzyme Activities of Inclusion Bodies of Recombinant ${\beta}$-Galactosidase via the Addition of Inducer Analog after L-Arabinose Induction in the araBAD Promoter System of Escherichia coli

  • Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.434-442
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    • 2008
  • We observed that an inclusion body (IB) of recombinant ${\beta}$-galactosidase that was produced by the araBAD promoter system in Escherichia coli (E. coil) showed enzyme activity. In order to improve its activity, the lowering of the transcription rate of the ${\beta}$-galactosidase structural gene was attempted through competition between an inducer (L-arabinose) and an inducer analog (D-fucose). In the deep-well microtiter plate culture and lab-scale fermentor culture, it was demonstrated that the addition of D-fucose caused an improvement in specific ${\beta}$-galactosidase production, although ${\beta}$-galactosidase was produced as an IB. In particular, the addition of D-fucose after induction led to an increase in the specific activity of ${\beta}$-galactosidase IB. Finally, we confirmed that the addition of D-fucose after induction caused changes in the structure of ${\beta}$-galactosidase IB, with higher enzyme activity. Based on these results, we expect that an improved enzyme IB will be used as a biocatalyst of the enzyme bioprocess, because an enzyme IB can be purified easily and has physical durability.

The outbreaks and surveillance of animal rabies in Gangwon-do

  • Shin Myung-Kyun;Shin Eun-Kyung;Chung Dong-Soo;Cheong Ki-Soo;Park Young-Nam;Choi Jee-Hee
    • Korean Journal of Veterinary Service
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    • v.27 no.4
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    • pp.335-343
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    • 2004
  • Although human rabies deaths are rare, the disease remains a public health problem in Korea. Here we report the outbreaks and surveillance of animal rabies in Gangwon-do. Animal rabies infections were identified in 119 animals from 1993 to 2003. The $78\%$ of all rabid animals were domestic species in Gangwon-do. Wild Korean raccoon dog (N. p. koreensis) continued to be the only reported rabid wildlife species. Outbreaks of rabies infections in Korean raccoon dogs are found in broad geographic regions across the northern Gangwon-do. The principal rabies hosts today are probably wild animals in Gangwon-do. The malaise, cerebral dysfunction, anxiety, confusion, agitation and abnormal behavior of the animals were the important symptoms of the disease. The Encephalitis, infiltration with lymphocytes and polymorphonuclear leukocytes and the inclusion bodies (Negri bodies) in neuronal cells were the specific histopathological signs. The results of indirect fluorescent antibody test (IFA) for animal rabies diagnosis were identical and the technique was useful to diagnose the disease. Preexposure vaccination is recommended for persons in high-risk groups, such as laboratory workers, veterinarians and certain animal handlers.

Fine Structure and Immunogoldlabeling of Crystalline Inclusion Bodies in Mitochondria (미토콘드리아내 결정함유물의 미세구조 및 면액황금표식법)

  • ;;R.A. Capaldi
    • The Korean Journal of Zoology
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    • v.31 no.1
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    • pp.62-70
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    • 1988
  • The distribution of respiratory chain complexes in beef heart and human muscle mitochondria has been explored by immunoeledron microscopy with antibodies made against beef heart mltochondriai proteins in conjundion with protein A cofloidai gold (l2nm particles). The antibodies used were made against NADH-conezyme Q reductase(complex I), ubiquinol-cytochrome-c-oxldoreductase (complex III) and cytochrome-c-oxidase(complex IV). Labeling of bed heart tissue with any of these antihodies gave gold particles randomly distributed along the mitochondrial inner membrane. The labeling of muscle tIssue mitochondria from a patient with a mitochondrial myopathy localized by biochemical analysis to complex III was quantitated and compared with the labeling of human control muscle tissue mitochondria. Four kinds of morphological changes in the mitochondrial fine strudure in the myopathy patient tissue have been found; paracrystalline inclusions consistIng of densely packed multi- lamellar structures, globular crystalline inclusions with high electron density, multilamellar strudure inclusion body(compadly and irregularly arranged concentric whirl shaped cristae)and golbular cyrstalilne inclusions located in the center of the whirl shaped cristae. Compex I and cytochrome-c-oxldase antihodies reacted to the same level in the mitochondria containing the crystalline inclusions and control mitochondria. Antibodies to complex III reacted very poorly to the mitochondria containing the crystalline Inclusions but strongly to control mitchondria. The globular crystalline inclusions in the mitochondria are not reacted antibodies to respiratory chain complexes.

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Production, Purification, and Characterization of Soluble NADH-Flavin Oxidoreductase(StyB) from Pseudomonas putida SN1

  • Yeo, Yun-Ji;Shin, Seung-Hee;Lee, Sun-Gu;Park, Sung-Hoon;Jeong, Yong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.362-367
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    • 2009
  • In recombinant strains, many proteins and enzymes are expressed as inactive and insoluble inclusion bodies. For soluble expression of an active form of StyB, an NADH-flavin oxidoreductase, several recombinant Escherichia coli strains were developed and tested. Among them, strain BL21(DE3)pLysS effectively produced an active and soluble form of StyB as about 9% of the total protein content, when cultivated at $20^{\circ}C$ with 0.5 mM IPTG. The solubly expressed StyB has the highest oxidoreductase activity at pH 6.5-7.5 and $37^{\circ}C$. Substrate dependence profiles of the StyB-catalyzed reaction showed that the maximum specific activity($V_m$) and half saturation constant($K_m$) were $1,867{\pm}148\;U/mg$ protein and $51.6{\pm}11{\mu}M$ for NADH, and $1,274{\pm}34\;U/mg$ protein and $8.2{\pm}1.2{\mu}M$ for FAD, respectively. This indicates that solubly produced StyB has 6- to 9-fold higher oxidoreductase activities than the in vitro refolded StyB from inclusion bodies.

In Vivo $^{13}C$-NMR Spectroscopic Study of Polyhydroxyalkanoic Acid Degradation Kinetics in Bacteria

  • Oh, Jung-Sook;Choi, Mun-Hwan;Yoon, Sung-Chul
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1330-1336
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    • 2005
  • Polyhydroxyalkanoic acid (PHA) inclusion bodies were analyzed in situ by $^{13}C$-nuclear magnetic resonance ($^{13}C$-NMR) spectroscopy. The PHA inclusion bodies studied were composed of poly(3-hydroxybutyrate) or poly(3hydroxybutyrate-co-4-hydroxybutyrate), which was accumulated in Hydrogenophaga pseudoflava, and medium-chain-length PHA (MCL-PHA), which was accumulated in Pseudomonas fluorescens BM07 from octanoic acid or 11-phenoxyundecanoic acid (11-POU). The quantification of the $^{13}C$-NMR signals was conducted against a standard compound, sodium 2,2-dimethyl-2-silapentane-5-sulfonate (DSS). The chemical shift values for the in vivo NMR spectral peaks agreed well with those for the corresponding purified PHA polymers. The intracellular degradation of the PHA inclusions by intracellular PHA depolymerase(s) was monitored by in vivo NMR spectroscopy and analyzed in terms of first-order reaction kinetics. The H. pseudoflava cells were washed for the degradation experiment, transferred to a degradation medium without a carbon source, but containing 1.0 g/l ammonium sulfate, and cultivated at $35^{\circ}C$ for 72 h. The in vivo NMR spectra were obtained at $70^{\circ}C$ for the short-chain-length PHA cells whereas the spectra for the aliphatic and aromatic MCL-PHA cells were obtained at $50^{\circ}C\;and\;80^{\circ}C$, respectively. For the H. pseudoflava cells, the in vivo NMR kinetics analysis of the PHA degradation resulted in a first-order degradation rate constant of 0.075/h ($r^{2}$=0.94) for the initial 24 h of degradation, which was close to the 0.050/h determined when using a gas chromatographic analysis of chloroform extracts of sulfuric acid/methanol reaction mixtures of dried whole cells. Accordingly, it is suggested that in vivo $^{13}C$-NMR spectroscopy is an important tool for studying intracellular PHA degradation in terms of kinetics.

A Novel Approach to Cloning and Expression of Human Thymidylate Synthase

  • Lv, Ying-Tao;Du, Pei-Juan;Wang, Qiao-Yan;Tan, Yuan;Sun, Zong-Bin;Su, Zhong-Liang;Kang, Cong-Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7523-7527
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    • 2013
  • Thymidylate synthase (TS) catalyzes the transfer of a methyl group from methylenetetrahydrofolate to dUMP to form dTMP. It is a primary target in the chemotherapy of colorectal cancers and some other neoplasms. In order to obtain pure protein for analysis of structure and biological function, an expression vector TS-pET28b (+) was constructed by inserting wild-type human thymidylate synthase (hTS) cDNA into pET28b (+). Then an expression strain was selected after transformation of the recombined plasmid into Rosetta (DE3). Fusion protein with His-tag was efficiently expressed in the form of inclusion bodies after IPTG induction and the content was approximately 40.0% of total bacteria proteins after optimizing expression conditions. When inclusion bodies were washed, dissolved and purified by Ni-NTA under denatured conditions, the purity was up to 90%. On SDS-PAGE and West-blotting, the protein band was found to match well with the predicted relative molecular mass-36kDa. Bioactivity was 0.1 U/mg. The results indicated that high-level expression of wild-type hTS cDNA can be achieved in prokaryotes with our novel method, facilitating research into related chemotherapy.

Expression and Characterization of Helicobacter pylori Adhesin Protein Linked to Cholera Toxin A2/B Subunits in Escherichia coli

  • Kim, Byung-Oh;Shin, Sung-Seup;Yoo, Young-Hyo;Pyo, Shuk-Neung
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.56-62
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    • 2000
  • The hpa gene genetically linked to the ctxa2b gene was cloned into the pTED expression vector, and the constructed pTEDhpa/ctxa2b was transformed into Excherichia coli. The fusion protein, the adhesin fused to the cholera toxin subunit A2B (CTXA2B) subunit, was expressed to high levels as inclusion bodies in E. coli. The expressed protein was partially purified by washing the inclusion bodies with working solution containing 8M Urea and 0.1M DTT. Refolding of denatured fusion protein was carried out in the presence of glutathione redox buffer. The refolded fusion protein was purified by size exclusion chromatography. The expressed fusion protein was verified by SDS-PAGE, western blotting with antibodies to both antigenic components of adhesin and cholera toxin subunit B (CTXB), and its N-terminal amino acid sequence was analyzed. The orderly assembled fusion protein was confirmed by modified Gm1-ganglioside ELISA with Abs to adhesin. The results indicate that the purified fusion protein is an Adhesin/CTXA2B protein containing the H. pylori adhesin and $G_{m1}4-ganglioside binding activity of CTXB and the expressed fusion protein in E. coli could be easily purified by the refolding process, Its molecular weight was 168kDa as estimated by size exclusion chromatography. The Adhesin/CTXA2B protein may be used as a candidate antigen for oral immunization against H. pylori.

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