• Title/Summary/Keyword: concentrated cells

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Large-Scale Culture of Hepatitis A Virus in Human Diploid MRC-5 Cells and Partial Purification of the Viral Antigen for Use as a Vaccine

  • Kim, Hyun-Seok;Chung, Yong-Ju;Jeon, Yeong-Joong;Lee, Sung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.386-392
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    • 1999
  • A large-scale culture of hepatitis A virus in human diploid MRC-5 cells was conducted. In a roller bottle culture, the virus was grown to a maximum titer in 3 weeks after infection. Over 95% of the cell-associated virus was excreted after culturing the infected cells in suspension media without fetal bovine serum for 3 days. The cultured virus was inactivated with formalin, concentrated by ultrafiltration, and partially purified by ultracentrifugation in a non-ionic gradient medium of Renocal. Two separate peak fractions showing high anti-HAY ELISA titer were pooled and about 40% of HAV antigen was recovered by this purification procedure. Of the partially purified vaccine, the protein pattern in SDS-PAGE and immunogenicity in mice were compared with a commercial HAV vaccine. In SDS-PAGE, the purified vaccine in this study and the commercial vaccine showed almost the same protein pattern. The seroconversion rate of the purified vaccine in mice was not different from that of the commercial vaccine. Therefore, we could prepare a good grade of HAV vaccine by a simple purification procedure although the purification itself was not completed.

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Possible Abnormalities of Chimeric Chicken Caused by the Introduction of Exogenous Genes Into Chicken Embryos via Primordial Germ Cells (PGCs)

  • Ebara, Fumio;Fujihara, Noboru
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.11
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    • pp.1514-1517
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    • 2000
  • In chicken, exogenous genes introduced into germinal crescent region (GCR) of the early developmental stage, where primordial germ cells (PGCs) were concentrated, were successfully transferred to the gonads via PGCs. The foreign genes were also confirmed to be successfully incorporated into F1 and F2 generations. We tried to incorporate the exogenous genes into PGCs by lipofection, then the DNA mixture was injected into GCR at stage 3-5 or 9-11 of embryonic development (Hamburger and Hamilton, 1951). The manipulated eggs were incubated, and hatched chicks were reared until sexual maturation. F1 generation was obtained from the DNA-treated chicken (DNA-chicken) mated with normal birds. Furthermore, F2 generation was also obtained from the F1 chicken mated with normal birds. The transfer of introduced foreign genes were confirmed by marker gene detection methods and PCR analysis in the hatched chicks, F1 and F2 generations. However, in our experiments, DNA-chickens showed abnormal characteristics such as low egg production rate, abnormal appearance and decreased number of spermatozoa. In the case of F1 chicken, low egg production and the deterioration of sperm capacity for insemination in male chicken were observed.

Nucleolar translocalization of GRA10 of Toxoplasma gondii transfectionally expressed in HeLa cells

  • Ahn, Hye-Jin;Kim, Sehra;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.45 no.3
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    • pp.165-174
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    • 2007
  • Toxoplasma gondii GRA10 expressed as a GFP-GRA10 fusion protein in HeLa cells moved to the nucleoli within the nucleus rapidly and entirely. GRA10 was concentrated specifically in the dense fibrillar component of the nucleolus morphologically by the overlap of GFP-GRA10 transfection image with IFA images by monoclonal antibodies against GRA10 (Tg378), B23 (nucleophosmin) and C23 (nucleolin). The nucleolar translocalization of GRA10 was caused by a putative nucleolar localizing sequence (NoLS) of GRA10. Interaction of GRA10 with TATA-binding protein associated factor 1B (TAF1B) in the yeast two-hybrid technique was confirmed by GST pull-down assay and immunoprecipitation assay. GRA10 and TAF1B were also co-localized in the nucleolus after co-transfection. The nucleolar condensation of GRA10 was affected by actinomycin D. Expressed GFP-GRA10 was evenly distributed over the nucleoplasm and the nucleolar locations remained as hollows in the nucleoplasm under a low dose of actinomycin D. Nucleolar localizing and interacting of GRA10 with TAF1B suggested the participation of GRA10 in rRNA synthesis of host cells to favor the parasitism of T. gondii.

Methanol Barriers Derived from Layer-by-Layer Assembly of Poly(ethersulfone)s for High Performance Direct Methanol Fuel Cells

  • Ok, Jung-Lim;Kim, Dong-Wook;Lee, Chang-Jin;Choi, Won-Choon;Cho, Sung-Min;Kang, Yong-Ku
    • Bulletin of the Korean Chemical Society
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    • v.29 no.4
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    • pp.842-846
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    • 2008
  • Layer-by-layer assembled multilayers of poly(ethersulfone)s were deposited on the surface of Nafion membrane for the application of direct methanol fuel cells (DMFC). Aminated poly(ethersulfone) (APES) and sulfonated poly(ethersulfone) (SPES) were used as a polycation and a polyanion for fabrication of the multilayer films. UV/Vis absorption spectroscopy verified a linear build-up of the multilayers of APES and SPES on the surface of Nafion. Thin multilayer films deposited on the Nafion membrane enabled methanol permeability of the membrane to decrease by 78% in comparison with the pristine Nafion. The performance of DMFCs in concentrated methanol was highly enhanced by using the multilayer modified Nafion.

Study for Improved Photocurrent via High Concentrated Tin-lead Perovskite Precursor Solution (주석-납 기반 페로브스카이트 고농도 전구체 용액을 이용한 광전류 향상 연구)

  • Hyojin Hong;Seungmin Lee;Jeong Min Im;Jun Hong Noh
    • Current Photovoltaic Research
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    • v.11 no.3
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    • pp.96-102
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    • 2023
  • Sn-Pb narrow-bandgap perovskite solar cells, which is a light-harvesting layer thicker than 1.3 micrometers, is needed to enhance the low photocurrent. The fabrication of such a thick film through solution processing is a key challenge. Here, we studied and characterized the film by using a precursor solution of increased concentration, comparing it with the universally used 1-micrometer Sn-Pb perovskite film. The increase in molar concentration clearly induced thickness enhancement, but we observed that it also created numerous voids at the interface with bottom charge transporting layer. We hypothesized that these voids might hinder the increase in photocurrent associated with thickness enhancement. By introducing methylammonium chloride (MACl), we successfully fabricated Sn-Pb perovskite film with a thickness of 1.3 micrometer and no voids. Void-controlled Sn-Pb perovskite solar cells not only demonstrated superior short-circuit current density compared to those with voids but also operated smoothly under light exposure.

The Role of Abp140p in Actin Dynamics of Budding Yeast

  • Lim, Bum-Soon;Lee, Yong-Keun;Pon, Liza A.;Yang, Hyeong-Cheol
    • International Journal of Oral Biology
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    • v.30 no.1
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    • pp.17-22
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    • 2005
  • In the previous studies of Saccharomyces cerevisiae, Abp140p (actin binding protein 140) fused to GFP has been only a protein that can label actin cables of yeast cells so far. However, the role of Abp140p in actin dynamics was remained elusive. In this study, the function of Abp140p was investigated with a deletion mutant and overexpression of GFP fused Abp140p. The deletion mutant was slightly more susceptible to Latrunculin-A (Lat-A), an actin-monomer sequestering agent, than wild type, although no significant deformation of actin structures was caused by ABP 140 deletion. Overexpression of Abp140p-GFP retarded cell growth, and produced thick and robust actin cables. Lat-A was not able to destabilize the thick actin cables, which suggests that actin dynamics was compromised in the cells with surplus of Abp140p. Therefore, Abp140p seems to stabilize actin cables together with other bundling proteins. Recently, actin cable dynamics of budding yeast was found to have a resemblance to that of filopodial tip of cultured mammalian cells. Retrograde movement of actin cables from buds to mother cells indicated local generation of the cable at bud sites. By using Abp140p-GFP, we traced the steps in the generation of a new actin cable after elimination of old cables by sodium azide. Before the appearance of a new actin cable, Abp140p-GFP concentrated in buds and disappeared, as mother cells became abundant in actin cables. Our observations provide a direct evidence of actin cable formation at buds of budding cells.

Homer2 regulates amylase secretion via physiological calcium oscillations in mouse parotid gland acinar cells

  • Kang, Namju;Kang, Jung Yun;Shin, Dong Min;Yang, Yu-Mi
    • International Journal of Oral Biology
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    • v.45 no.2
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    • pp.58-63
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    • 2020
  • The salivary glands secrete saliva, which plays a role in the maintenance of a healthy oral environment. Under physiological conditions, saliva secretion within the acinar cells of the gland is regulated by stimulation of specific calcium (Ca2+) signaling mechanisms such as increases in the intracellular Ca2+ concentration ([Ca2+]i) via storeoperated Ca2+ entry, which involves components such as Orai1, transient receptor potential (TRP) canonical 1, stromal interaction molecules, and inositol 1,4,5-triphosphate (IP3) receptors (IP3Rs). Homer proteins are scaffold proteins that bind to G protein-coupled receptors, IP3Rs, ryanodine receptors, and TRP channels. However, their exact role in Ca2+ signaling in the salivary glands remains unknown. In the present study, we investigated the role of Homer2 in Ca2+ signaling and saliva secretion in parotid gland acinar cells under physiological conditions. Deletion of Homer2 (Homer2-/-) markedly decreased the amplitude of [Ca2+]i oscillations via the stimulation of carbachol, which is physiologically concentrated in parotid acinar cells, whereas the frequency of [Ca2+]i oscillations showed no difference between wild-type and Homer2-/- mice. Homer2-/- mice also showed a significant decrease in amylase release by carbachol in the parotid gland in a dose-dependent manner. These results suggest that Homer2 plays a critical role in maintaining [Ca2+]i concentration and secretion of saliva in mouse parotid gland acinar cells.

A Study on the Gymnodinium nagasakiense Red-Tide in Jinhae Bay of Korea (진해만의 Gymnodinium nagasakiense 적조에 관한 연구)

  • Lee, Jin-Hwan;Kwak, Hi-Sang
    • The Korean Journal of Ecology
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    • v.9 no.3
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    • pp.149-160
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    • 1986
  • Relationship between the causative organi는 of red-tide and environmental factors had been ecologically dealt wtih. The surveys were conducted at seven station in Jinhae Bay from July to September 1981. The water temperature and salinity had wide range, i.e. 23.3~29.3$^{\circ}C$ and 19G.78~31.29$\textperthousand$, but several chemical factors remarkably fluctuated; dissolved oxygen 102.9~210.4%, COD 2.10~8.96mg$O_2$/l, pH 8.1~9.4, $NO_3$-N trace~1, 052$\mu$g/l, $PO_4$-P 0.6~58.9$\mu$g/l and chlorophyll-a 2.18~290.5mg/$m^3$ in the observed area. The red-tide was mainly caused by two dinoflagellata taxa throughout major outbreaks occurred in July through September. Leading species of red-tide were Gymnodinium nagasakiense belong to the ajor species. During the surveyed period, cell nubers of the causative organisms of the red-tide extensively varied from 3${\times} /10^4$ cells/l to $1.5\times10^7 $cells/l with moths and stations; Prorocentrum spp. 0.3~12.5$\times\10^5$ cells/l in July; Gymnodinium nagasakiense 0.2~5.9$\times10^6 cells/l, 1.1~4.7$\times10^6$ cells/l, and 0.2~15.1$\times10^6$ cells/l in July, August, and September, respectively. Gymnodinium nagasakiense red-tide seemed to be caused by the high water temperature in summer, unusually low salinity due to heavy rains, and the concentrated nutrients for phytoplankton supplied with the municipal sewages from the urban areas and the wastewaters from the industrial complexes.

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Study on Distribution of Oct4 Expression and Change of Apoptosis in Nuclear Transfer Blastocyst using Oct4-Transfected Mesenchymal Stem Cells (Oct4-Transfection한 중간엽줄기세포 유래 핵이식 배반포의 Oct4 발현 분포 및 세포 자멸사의 변화에 관한 연구)

  • Lee, Won-Jae;Lee, Jeong-Hyeon;Rho, Gyu-Jin;Lee, Sung-Lim
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.81-88
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    • 2016
  • There are various factors i.e. donor cell type, culture system as well as technical procedures which influence the pre-implantation embryonic development; however, may attempts have been made and still it is under investigation to improve the cloning efficiency using somatic cell nuclear transfer technique. It is has been investigated that stem cells like mesenchymal stem cell are able to more efficiently reprogram and reactivate the expression of early embryonic genes to promote nuclear transfer efficiency. In addition, Oct4 expression plays a pivotal role in early embryo development. In the present study, we investigated distribution of Oct4 expression and changes of apoptosis and total cell number in nuclear transfer blastocyst after using Oct4 transfected bone marrow stem cell as donor cells. Although Oct4-RFP expression was observed across blastocyst, more concentrated intensity was shown at hatched region in blastocyst on day 7. Reduction of apoptotic bodies was revealed in Oct4 transfected blastocyst by TUNEL staining, however, there was no significant difference in total cell number between Oct4 transfected and non-transfected nuclear transfer embryos. In conclusion, Oct4 transfected donor cells exhibited higher expression in hatching sight in day 7 blastocyst and were able to prevent apoptosis compared to non-transfected donor cells.

Anti-inflammation and Anti-inflammasome Effects of Bambusae Caulis in Liquamen mediated by Nrf2 Activation in Kupffer cells (쿠퍼 세포에서 Nrf2 활성화 매개 죽력의 염증 및 인플라마좀 억제 효능)

  • Ji Hye Yang
    • Herbal Formula Science
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    • v.31 no.4
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    • pp.253-264
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    • 2023
  • Objectives : Bambusae Caulis in Liquamen (BCL), a traditional herbal medicine, is a distilled product of condensation from the burning of fresh bamboo stems. We previously identified the anti-oxidant capacity of BCL in hepatocytes and suggested that BCL is a promising therapeutic candidate for treating oxidative stress-induced hepatocellular damage. Despite the importance of the role played by Kupffer cells in liver disease, the efficacy of BCL on Kupffer cells is unclear. Therefore, this study aimed to determine whether BCL could suppress LPS-induced inflammation and LPS+ATP-induced inflammasomes in Kupffer cells. Methods : We used ImKCs, a murine immortalized Kupffer cell line to examined whether BCL inhibited LPS-induced inflammation response and oxidave stress. And, we prepared a total of 18 L of BCL, purchased from Bamboo Forest Foods Co., Ltd. (648 Samdari, Damyang-eup, Damyang-gun, Jeollanam-do, Republic of Korea), was concentrated using a decompression concentrator. Result : The LPS-induced release of inflammatory cytokines was abolished by BCL treatment. Also, BCL treatment suppressed the LPS+ATP-induced expression of inflammasome proteins (NLRP3, IL-1, and IL-18), and inhib β ited the release of IL-1 . BCL decreased LPS-or LPS+ATP-induc β ed reactive oxygen species production. In addition, BCL increased nuclear translocation of Nrf2 and the expression of HO-1 in a time-dependent manner. Conclusion : These results suggest the efficacy of BCL with respect to its anti-inflammatory and anti-inflammasome effects mediated by Nrf2 in Kupffer cells.