• Title/Summary/Keyword: pre-B cell

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Label-free Noninvasive Characterization of Osteoclast Differentiation Using Raman Spectroscopy Coupled with Multivariate Analysis

  • Jung, Gyeong Bok;Kang, In Soon;Lee, Young Ju;Kim, Dohyun;Park, Hun-Kuk;Lee, Gi-Ja;Kim, Chaekyun
    • Current Optics and Photonics
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    • v.1 no.4
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    • pp.412-420
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    • 2017
  • Multinucleated bone resorptive osteoclasts differentiate from bone marrow-derived monocyte/macrophage precursor cells. During osteoclast differentiation, mononuclear pre-osteoclasts change their morphology and biochemical characteristics. In this study, Raman spectroscopy with multivariate techniques such as Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA) were used to extract biochemical information related to various cellular events during osteoclastogenesis. This technique allowed for label-free and noninvasive monitoring of differentiating cells, and clearly discriminated four different time points during osteoclast differentiation. The Raman band intensity showed significant time-dependent changes that increased up to day 4. The results of Raman spectroscopy agreed with results from atomic force microscopy (AFM) and tartrate-resistant acid phosphatase (TRAP) staining, a conventional biological assay. Under AFM, normal spindle-like mononuclear pre-osteoclasts became round and smaller at day 2 after treatment with a receptor activator of nuclear $factor-{\kappa}B$ ligand and they formed multinucleated giant cells at day 4. Thus, Raman spectroscopy, in combination with PCA-LDA, may be useful for noninvasive label-free quality assessment of cell status during osteoclast differentiation, enabling more efficient optimization of the bioprocesses.

Anti-Inflammmatiry Effects of Nerium indicum Ethanol Extracts through Suppression of NF-kappaB Activation (NF-κB 활성 저해를 통한 협죽도 에탄올 추출물의 항염증 효능)

  • Kim, Tae-Hwan;Ko, Seog-Soon;Park, Cheol;Park, Sang-Eun;Hong, Sang-Hoon;Kim, Byung-Woo;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1221-1229
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    • 2010
  • Nerium indicum, an India-Pakistan-originated shrub belonging to the oleander family, is reported to possess many pharmacological activities including cardiac muscle stimulation, and anti-diabetes, anti-angiogenesis, anti-cancer and neuro-protective activities. However, the anti-inflammatory properties of N. indicum were unclear. In this study, we investigated the effects of ethanol extract of the N. indicum leaf and stem (ENIL and ENIS) on the expression of anti-inflammatory mediators in U937 human pre-monocytic cell models. In U937 cells stimulated with phorbol 12-myristate-13-acetate (PMA), pre-treatment with ENIS significantly inhibited the expression of both cyclooxygenase-2 (COX-2) mRNA and protein, which are associated with inhibition of the release of prostaglandin $E_2\;(PGE_2)$, whereas the inhibitory effects appeared weakly in ENIL. Moreover, ENIS significantly attenuated PMA-induced IkappaB ($I{\kappa}B$) degradation and suppressed elevated nuclear factor kappa B (NF-${\kappa}B$) nuclear translocation. Taken together, these findings provide important new insights that N. indicum exhibits anti-inflammatory properties by suppressing the transcription of pro-inflammatory cytokine genes through the NF-kB signaling pathway.

Anti-inflammatory effects of Sparassis crispa extracts (꽃송이버섯 추출물의 항염활성 효과)

  • Choi, Woo-Suk;Shin, Pyung-Gyun;Bok, Yoo Young;Jun, Noh Hyung;Kim, Gun-Do
    • Journal of Mushroom
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    • v.11 no.1
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    • pp.46-51
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    • 2013
  • Sparassis crispa is a medicinal mushroom, which has been reported to have anti-cancer effect. In this study, we designed to investigate the effects of Sparassis crispa extracts on the production of nitric oxide (NO) in LPS-stimulated RAW264.7 cells. The pre-treatment of the extracts prior to add LPS in RAW264.7 cells suppressed NO production and iNOS expression at protein and mRNA levels. The phosphorylation of $I{\kappa}B{\alpha}$ was inhibited by the extracts, which was induced through suppressing the activation of $NF-{\kappa}B$. Sparassis crispa extracts showed the effect on the down-regulation of STAT-1 activation in a dose-dependent manner. In LPS-stimulated RAW264.7 cells, $NF-{\kappa}B$ was translocated into the nucleus, while the treatment of Sparassis crispa extracts induced to sequestered $NF-{\kappa}B$ in the cytosol. These experimental results determined that Sparassis crispa extracts play a inhibitory role in inflammatory reactions via regulating NO production, which suggests potential as a component of inflammatory drugs.

Prevention of Quality Changes in the Cultured Wild Ginseng During Storage (산삼배양근의 저장 중 품질변화 억제)

  • Whang, Jong-Hyun;Yu, Kwang-Won;Park, Sung-Sun;Koh, Jong-Ho;Oh, Sung-Hoon;Suh, Hyung-Joo;Lee, Sang-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1312-1317
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    • 2008
  • Physicochemical changes were investigated for the shelf-life extension of cultured wild-ginseng roots during storage with various pre-treatments with blanching, CAMICA-SD and DF-100 and treatments with citric acid and vitamin C. The pH of cultured wild-ginseng roots showed the range of $6.06{\sim}6.42$ at $10^{\circ}C$, but showed higher ranges of $6.08{\sim}6.91$ and $6.08{\sim}8.68$ at 20 and $30^{\circ}C$, respectively. Browning index (a/b) was increased with increasing storage temperature, and the index at 10 and $30^{\circ}C$ were 0.405 and 0.469 after 2 weeks, respectively. Browning index and viable cell number of CAMICA-SD pre-treatment showed little changes compared to pre-teatment with blanching or DF-100. When the cultured wild-ginseng roots were treated with 1.0% citric acid and 0.2% DF-100 after pre-treatments with CAMICA-SD, viable cell number was slightly increased to $4.9{\times}10^2CFU/g$ for 3 weeks storage at $10^{\circ}C$. The mixture of citric acid and DF-100 was also used to prevent the growth of microbiology and to reduce browning reaction, especially enzymatic browning reaction. The mixture might effectively extend shelf life of the cultured wild-ginseng roots.

Post-Activation Treatment with Cytochalasins and Latrunculin A on the Development of Pig Oocytes after Parthenogenesis and Somatic Cell Nuclear Transfer

  • Park, Bola;Lee, Joohyeong;Lee, Yongjin;Elahi, Fazle;Jeon, Yubyeol;Hyun, Sang-Hwan;Lee, Eunsong
    • Journal of Embryo Transfer
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    • v.28 no.2
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    • pp.133-139
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    • 2013
  • The objective of this study was to determine the effect of post-activation treatment with cytoskeletal regulators in combination with or without 6-dimethylaminopurine (DMAP) on embryonic development of pig oocytes after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT). PA and SCNT oocytes were produced by using in vitro-matured pig oocytes and treated for 4 h after electric activation with $0.5{\mu}M$ latrunculin A (LA), $10.4{\mu}M$ cytochalasins B (CB), and $4.9{\mu}M$ cytochalasins D (CD) together with none or 2 mM DMAP. Post-activation treatment of PA oocytes with LA, CB, and CD did not alter embryo cleavage (85.8~88.6%), blastocyst formation (30.7~ 32.4%), and mean cell number of blastocysts (33.5~33.8 cells/blastocyst). When PA oocytes were treated with LA, CB, and CD in combination with DMAP, blastocyst formation was significantly (P<0.05) improved by CB+DMAP (42.5%) compared to LA+DMAP (28.0%) and CD+DMAP (25.1%), but no significant differences were found in embryo cleavage (77.5~78.0%) and mean blastocyst cell number (33.6~35.0 cells) among the three groups. In SCNT, blastocyst formation was significantly (P<0.05) increased by post-activation treatment with LA+DMAP (32.9%) and CD+DMAP (35.0%) compared to CB+DMAP (22.0%) while embryo cleavage (85.5~85.7%) and blastocyst cell number (41.1~43.8 cells) were not influenced. All three treatments (LA, CB, and CD with DMAP) effectively inhibited pseudo-polar body extrusion in SCNT oocytes. The proportions of oocytes showing single pronucleus formation were 89.6%, 83.9%, and 93.3%, respectively with the increased tendency (P<0.1) by LA+DMAP and CD+ DMAP compared to CB+DMAP. Our results demonstrate that post-activation treatment with LA or CD in combination with DMAP improves pre-implantation development of SCNT embryos and the stimulating effect of cytoskeletal modifiers on embryonic development is differentially shown depending on the origin (PA or SCNT) of embryos in pigs.

Bacterial determinants involved in the induction of systemic resistance ana plant growth promotion in tobacco by Pseudomonas chlororaphis O6.

  • Han, Song-Hee;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.101.2-102
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    • 2003
  • The ability of P. chlororaphis O6 to induce resistance to Erwinia carotovora subsp. carotovara SCCI and to promote growth in tobacco was demonstrated in microtiter assays on plants pre-inoculated at the root level with the bacteria before challenge with the leaf pathogen. To identify th bacterial determinants involved in induced systemic resistance and plant growth promotion, cell culture of O6 grown in King's medium B was fractionated with organic solvents and purified using various columns. in vivo and in vitro assays with samples from successive fractionation steps of the O6 supernatant led to the conclusion that antibacterial compounds were observed in aqueous layer, and to the isolation of fractions containing metabolites that retained most of the resistance-inducing activity (70:30, methanol:water) and the plant growth promotion (80:20 and 90:10, methanol:water) after ODS column chromatography. Although these molecules remain to be purified further and structurally characterized, its isolation is an addition to the range of determinants from plant growth-promoting rhizobacteria known to stimulate plant defence.

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Crystal Structure of Rattus norvegicus Visfatin/PBEF/Nampt in Complex with an FK866-Based Inhibitor

  • Kang, Gil Bu;Bae, Man-Ho;Kim, Mun-Kyoung;Im, Isak;Kim, Yong-Chul;Eom, Soo Hyun
    • Molecules and Cells
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    • v.27 no.6
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    • pp.667-671
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    • 2009
  • Visfatin (Nampt/PBEF) plays a pivotal role in the salvage pathway for $NAD^+$ biosynthesis. Its potent inhibitor, FK866, causes cellular $NAD^+$ levels to decline, thereby inducing apoptosis in tumor cells. In an effort to improve the solubility and binding interactions of FK866, we designed and synthesized IS001, in which a ribose group is attached to the FK866 pyridyl ring. Here, we report the crystal structure of rat visfatin in complex with IS001. Like FK866, IS001 is positioned at the dimer interface, and all of the residues that interact with IS001 are involved in hydrophobic or ${\pi}-{\pi}$-stacking interactions. However, we were unable to detect any strong interactions between the added ribose ring of IS001 and visfatin, which implies that a bulkier modifying group is necessary for a tight interaction. This study provides additional structure-based information needed to optimize the design of visfatin inhibitors.

cDNA Sequences for Asialoglycoprotein Receptor from Human Fetal Liver

  • Lee, Dong-Gun;Lee, Sung-Gu;Kim, Kil-Lyong;Hahm, Kyung-Soo
    • BMB Reports
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    • v.30 no.4
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    • pp.299-301
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    • 1997
  • The asialoglycoprotein receptor (ASGPR) was the first described mammalian lectin that mediates the specific binding and internalization of galactose/N-acetylgalactosamine-terminating glycoproteins by hepatic parenchymal cells. H1 and H2 are known as essential subunits of the functional ASGPR. There were close similarities in ASGPR H2 subunits between cultured cell line HepG2 and normal human liver cells including identical sequences at both termini. It was therefore expected that there may be some similarities between the subunits from normal liver cells and fetal liver cells. The two subunits of human fetal liver ASGPR. designated FL-H1 and FL-H2. were cloned from cDNA library by peR and the sequences were compared with the known HI and H2 sequences of HepG2, and the H1 sequence of nornal human liver cells. The results showed that FL-H1 was identical to H1 of HepG2. Whereas FL-H2 contains a 15-bp miniexon, but missing 57-bp at the near upstream from the membrane-spanning domain compared to H2 of HepG2 and normal human liver cells indicating that FL-H2 resulted from a differential splicing compared to HepG2 and normal liver cells.

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Effects of Sleep Promoting Program on Sleep and Immune Response in Elderly (수면증진 프로그램이 노인의 수면과 면역반응에 미치는 효과)

  • Hong, Se-Hoon;Kim, Sook-Young
    • Korean Journal of Adult Nursing
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    • v.21 no.2
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    • pp.167-178
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    • 2009
  • Purpose: This study was to examine the effects of sleep promoting program on sleep and the immune response in the elderly with insomnia. Methods: The study was designed as a quasi-experimental, nonequivalent control group pre-post test design. Thirty two elders who suffered from insomnia (16 experimental and 16 control subjects) were selected among the elders who had been enrolled in two churches. The subjects in the experimental group participated in a sleep promoting program which was composed of sleep hygiene education and progressive muscle relaxation for 4 weeks. Data were analyzed using the SPSS/WIN. Results: The experimental group showed higher sleep score than that of the control group (t=7.86, p=.00). The experimental group showed higher sleep satisfaction score than that of the control group (t=7.61, p=.00). The sleep promoting program was not effective in enhancing immune response. Conclusion: The sleep promoting program increased the sleep score, sleep-satisfaction score and B cell in immune response of elderly people suffering from insomnia. Therefore, sleep promoting program can be applied as an effective nursing intervention to promote sleep quality and sleep satisfaction.

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Post-transcriptional and post-translational regulation during mouse oocyte maturation

  • Kang, Min-Kook;Han, Seung-Jin
    • BMB Reports
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    • v.44 no.3
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    • pp.147-157
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    • 2011
  • The meiotic process from the primordial stage to zygote in female germ cells is mainly adjusted by post-transcriptional regulation of pre-existing maternal mRNA and post-translational modification of proteins. Several key proteins such as the cell cycle regulator, Cdk1/cyclin B, are post-translationally modified for precise control of meiotic progression. The second messenger (cAMP), kinases (PKA, Akt, MAPK, Aurora A, CaMK II, etc), phosphatases (Cdc25, Cdc14), and other proteins (G-protein coupled receptor, phosphodiesterase) are directly or indirectly involved in this process. Many proteins, such as CPEB, maskin, eIF4E, eIF4G, 4E-BP, and 4E-T, post-transcriptionally regulate mRNA via binding to the cap structure at the 5' end of mRNA or its 3' untranslated region (UTR) to generate a closed-loop structure. The 3' UTR of the transcript is also implicated in post-transcriptional regulation through an association with proteins such as CPEB, CPSF, GLD-2, PARN, and Dazl to modulate poly(A) tail length. RNA interfering is a new regulatory mechanism of the amount of mRNA in the mouse oocyte. This review summarizes information about post-transcriptional and post-translational regulation during mouse oocyte meiotic maturation.