• Title/Summary/Keyword: concentrated cells

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Protoberberine alkaloids from the rhizome of Coptis japonica Makino

  • Min , Yong-Deuk;Chung, Ae-Kyung;Shim, Sang-Ho;Lim, Bang-Ho;Shin, Dae-Hee;Lee, Kang-Ro
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.205.2-205.2
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    • 2003
  • As part of our program to isolate bioactive compounds from korean natural sources, we have screened traditional medicinal plants to cytotoxity on human tumor cells. Of them, the MeOH extract from rhizome of Coptis japonica Makino was found to be active against five cultured human tumor cell lines. So, the MeOH extract was subjected to successive solvent partitioning to give n-hexane, chloroform and BuOH. The activity was concentrated into the chloroform extract. The extract was chromatographied on a silica gel column and resulted in the isolation 5 alkaloids. (omitted)

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Anti-inflammatory Effects of Naetakbaekryeom-san (내탁백렴산 추출물의 항염증 효능 연구)

  • Jung, Min Jae;Noh, Hui Jeong;Choi, Ji Min;Jeon, Seok Hee;Kim, Seon Jong
    • Journal of Korean Medicine Rehabilitation
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    • v.32 no.4
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    • pp.9-18
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    • 2022
  • Objectives This study was conducted to confirm the anti-inflammatory effect of Naetakbaekryeom-san (NTB), and whether it could be another treatment for inflammatory diseases. Methods The NTB water extract was extracted with hot water at 100℃ for 2 hours, concentrated at 80℃ under reduced pressure, and used. After 2 hours of pretreatment with NTB and positive control Bay11-7082, nitric oxide (NO), inducible NO synthase (iNOS), interleukin (IL)-6, IL-1𝛽, tumor necrosis factor alpha (TNF-𝛼) were measured in RAW264.7 cells activated with lipopolysaccharides (LPS) 500 ng/mL. After 2 hours of pretreatment with NTB, the anti-inflammatory effect of NTB was evaluated by measuring nuclear factor kappa-light-chain-enhancer of activated B cells (NF-𝜅B) in RAW264.7 cells and 293T cells activated with phorbol 12-myristate 13-acetic acid (PMA) 30 ng/mL. Results In RAW264.7 cells activated with LPS, NTB at concentrations of 0.1, 0.3, and 1.0 mg/mL showed no cytotoxicity, significantly inhibited NO production and inhibition of iNOS expression. TNF-𝛼 cytokine levels was not regulated, but NTB at each concentration inhibited the production of IL-1𝛽 and IL-6, and the effect was higher than that of the positive control Bay11-7082 (20 𝜇M). In PMA-activated RAW264.7 cells and 293T cells, each concentration of NBT decreased the NF-𝜅B transcriptional activity, with the greatest decrease at 1 mg/mL. Conclusions These results demonstrated the anti-inflammatory effect of NTB water extracts, but further studies such as comparison of anti-inflammatory effects and antioxidant effects by NTB component, comparison of effects according to extraction solvents, and clinical studies are needed.

The Disruption of Saccharomyces cerevisiae Cells and Release of Glucose 6-Phosphate Dehydrogenase (G6PDH) in a Horizontal Dyno Bead Mill Operated in Continuous Recycling Mode

  • Mei Chow Yen;Ti Tey Beng;Ibrahim Mohammad Nordin;Ariff Arbakariya;Chuan Ling Tau
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.284-288
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    • 2005
  • Baker's yeast was disrupted in a 1.4-L stainless steel horizontal bead mill under a continuous recycle mode using 0.3 mm diameter zirconia beads as abrasive. A single pass in continuous mode bead mill operation liberates half of the maximally released protein. The maximum total protein release can only be achieved after passaging the cells 5 times through the disruption chamber. The degree of cell disruption was increased with the increase in feeding rate, but the total protein release was highest at the middle range of feeding rate (45 L/h). The total protein release was increased with an increase in biomass concentration from 10 to $50\%$(w/v). However, higher heat dissipation as a result of high viscosity of concentrated biomass led to the denaturation of labile protein such as glucose 6-phosphate dehydrogenase (G6PDH). As a result the highest specific activity of G6PDH was achieved at biomass concentration of $20\%$(ww/v). Generally, the degree of cell disruption and total protein released were increased with an increase in impeller tip speed, but the specific activity of G6PDH was decreased substantially at higher impeller tip speed (14 m/s). Both the degree of cell disruption and total protein release increased, as the bead loading increased from 75 to $85\% (v/v)$. Hence, in order to obtain a higher yield of labile protein such as G6PDH, the yeast cell should not be disrupted at biomass concentration and impeller tip speed higher than $20\%(w/v)$ and 10 m/s, respectively.

Inhibitory Effect of Oriental Herbal Medicines on Tumor Necrosis $Factor-{\alpha}$ Production in Lipopolysaccharide-stimulated RAW264.7 cells

  • Cho, Jae-Youl;Park, Ji-Soo;Kim, Pyung-Su;Chae, Sook-Hee;Yoo, Eun-Sook;Baik, Kyong-Up;Lee, Jong-Soo;Park, Myung-Hwan
    • Natural Product Sciences
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    • v.5 no.1
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    • pp.12-19
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    • 1999
  • Eleven out of 118 herbal medicines which are frequently used in Korean traditional prescriptions for inflammatory diseases exhibited more than 50% of inhibition on $TNF-{\alpha}$ production in LPS-stimulated RAW264.7 cells by their total ethanol extracts with 0.1 mg/ml as a final concentration. The active 11 total extracts were prepared from Angelica koreana, Coptis japonica, Cynanchum paniculatum, Magnolia frgesii, Magnolia officinalis, Panax ginseng, Patrinia scabiosaefolia, Pterocarpus santalius, Rhapontica uniflora, Saussurea lappa. Of them, Coptis japonica, Magnolia fargesii and Saussurea lappa also significantly inhibited $TNF-{\alpha}$ production in vivo. These total extracts were sequentially fractionated with n-hexane, methylene chloride, ethyl acetate, n-butanol and water. Among the solvent-fractionated extracts with 0.05 mg/ml as a final concentration, inhibitory effects of Angelica koreana, Magnolia fargesii, Magnolia officinalis, Pterocarpus santalinus, Rhapontica uniflora and Saussurea lappa markedly showed in one or two solvent fractions suggesting that the principles may be concentrated by subfractionation as the main compounds.

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Anti-Wrinkling Effect of Noni (Morinda citrifolia) by Antioxidant and Anti-Inflammatory Properties

  • Choi, Soo-Cheol;Youn, Young Han
    • Journal of People, Plants, and Environment
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    • v.23 no.2
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    • pp.191-199
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    • 2020
  • Noni has been used for medicinal purposes for more than 2,000 years in South Pacific Polynesia, China and India, and has been heavily ingested as an extract for its excellent antioxidant and anti-inflammatory effects. However, a recent study found that the noni extract causes digestive disorders, kidney problems, and liver diseases, which made it necessary to use it for other purposes than as an extract. In this study, we want to evaluate the potential of noni as an anti-oxidant, anti-inflammatory and anti-wrinkling agent. Methods: Noni was freeze-dried, extracted in water, and concentrated. Skin cells were treated with the noni extract for 24 hrs and then were exposed to UVB (55 mJ/cm2). After 48 hrs of incubation, pro-inflammatory cytokine, elastase, MMP-1 and type-1 procollagen levels were measured by ELISA. Results: To find out the antioxidant effect of the noni extract, the DPPH and ABTS radical scavenging activity experiments were conducted and the noni extract showed 97.0 % and 92.0 % antioxidant efficacy at 200 ㎍/mL respectively. The noni extract (50 and 100 ㎍/mL) decreased IL-6 and TNF-α in RAW 264.7 cells induced by LPS in a concentration-dependent manner. In the RT-PCR experiment involving NO production, the noni extract (50 and 100 ㎍/mL) inhibited NO production by strongly inhibiting iNOS mRNA expression, and also inhibited the elevation of MMP-1 and elastases caused by UVB irradiation by 25.0 % and 7.0 % respectively. In addition, type-1 procollagen was elevated by 20.0 % by the noni extract treatment in HaCaT cells. Conclusion: The noni extract has photoprotective ability by reducing proinflammatory mediators, elastase and MMP-1 production, and elevation of collagen synthesis. Our findings suggest that the noni extract might be a good natural substance to protect against UVB-induced premature skin aging.

Cytokine Storm Related to CD4+ T Cells in Influenza Virus-Associated Acute Necrotizing Encephalopathy

  • Shushu Wang;Dongyao Wang;Xuesong Wang;Mingwu Chen;Yanshi Wang;Haoquan Zhou;Yonggang Zhou;Yong Lv;Haiming Wei
    • IMMUNE NETWORK
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    • v.24 no.2
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    • pp.18.1-18.12
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    • 2024
  • Acute necrotizing encephalopathy (ANE) is a rare but deadly complication with an unclear pathogenesis. We aimed to elucidate the immune characteristics of H1N1 influenza virus-associated ANE (IANE) and provide a potential therapeutic approach for IANE. Seven pediatric cases from a concentrated outbreak of H1N1 influenza were included in this study. The patients' CD4+ T cells from peripheral blood decreased sharply in number but highly expressed Eomesodermin (Eomes), CD69 and PD-1, companied with extremely high levels of IL-6, IL-8 in the cerebrospinal fluid and plasma. Patient 2, who showed high fever and seizures and was admitted to the hospital very early in the disease course, received intravenous tocilizumab and subsequently showed a reduction in temperature and a stable conscious state 24 h later. In conclusion, a proinflammatory cytokine storm associated with activated CD4+ T cells may cause severe brain pathology in IANE. Tocilizumab may be helpful in treating IANE.

Antigenic localities in the tissues of Payagonimus westermani by developmental stages using immunogoldlabeling method (면역황금표식법을 이용한 폐흡충의 발육단계별 충체조직내 항원성 부위에 관한 연구)

  • 임한종;김수진
    • Parasites, Hosts and Diseases
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    • v.30 no.1
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    • pp.1-14
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    • 1992
  • In order to observe the antigenic localization in the tissues of Paragonimus westermani of deve- lopmental stages, immunogoldlabeling method was applied using serum of the cats which were infected with isolated metacercariae from Cambaroides similis. The sectioned worm tissues from orch developmental stage were embedded in Lowicryl HM20 medium, stained with infected semi IgG and protein A gold complex(particle size: 12 nm) and observed by electron microscopy. In the young adult worm tissue of 4 weeks after infection with metacercariae, the gold particles were specifically concentrated on the tegumental syncytium and cytoplasm of the tegumental cells as well as the secretory granules in the parenchymal tissue. The antigenic materials in the adult worm tissue were specifically concentrated on the secretory granules in the parenchymal tissue, the cytoplasm between granules in the vitelline gland and the epithelial lamella in the lumen of the caecum.

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An Efficient Method for the Release of Recombinant Penicillin G Amidase from the Escherichia coli Periplasm (대장균의 periplasm으로부터 재조합 PGA 단백질의 효율적이고 간단한 방출 방법)

  • Lee, Sang-Mahn
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1145-1151
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    • 2017
  • In this study, we report on a simple, efficient method for obtaining penicillin G amidase (PGA) from recombinant Escherichia coli using a formulation mixed with detergent and lysozyme. Research was conducted on the extraction efficiency of PGA from the periplasmic space in cells in terms of the type of detergent, detergent concentration, pH, reaction time, and temperature of permeabilization. The extraction yield of PGA in the formulated surfactant/lysozyme treatment was increased by approximately (55-65 U/ml) in comparison with that in the single surfactant treatment. The released PGA solution was concentrated and exchanged with buffer using an ultrafiltration (U/F) system. The yields of diatomite filtration, membrane filtration (M/F), and U/F were 69.7%, 93.8%, and 77.3%, respectively. A total of 212 KU of PGA was recovered. At the 25-L culture scale, the overall yield of extraction using the mixed surfactant/lysozyme method was 49.2%. The specific activity of extracted PGA was 11 U/mg in protein. The concentrated PGA solution was immobilized on microporous silica beads without further purification of PGA. The total immobilization yield of PGA on the resin was 48.7%, while the enzyme activity was 101 U/g. The immobilized PGA was successfully used to produce 6-APA from penicillin G. Our results indicated that a simple extraction method from periplasmic space in E. coli may be used for the commercial scale production of ${\beta}-lactam$ antibiotics using immobilized PGA.

The Effect of Bee Venom Pharmacopuncture Therapy on the Condition of Different Concentration in Rheumatoid Arthritis Rat Model (흰쥐의 류마티스 관절염 모델에서 봉약침의 농도별 처리 조건에 따른 치료 효과)

  • You, Deok-Seon;Yeom, Seung-Ryong;Lee, Su-Kyung;Kwon, Young-Dal;Song, Yung-Sun
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.2
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    • pp.101-123
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    • 2011
  • Objectives : The aim was to study the effect of bee venom pharmacopuncture therapy with different concentration on rheumatoid arthritis rat model. Methods : We enforced a bee venom pharmacopuncture therapy with different concentration on rheumatoid arthritis rat model by the intradermal injection of chicken type II collagen emulsified. 14 days after the onset of the rheumatoid arthritis rat model, a fixed volume of bee venom was daily injected to ST-35 acupoint in the rat's knee joint for 2-3 weeks. The hind paw volume, arthritic index, arthritic flexion pain test, pain threshold, and serum analysis (CRP, $PGE_2$, ALT, AST) were analyzed, and the expression profiles of COX-2, c-fos, and substance-P at the dorsal horn region of the spinal cord and subchondral bone of the knee joint were also analyzed by using the immunohistochemistry. Results : After the treatment of rheumatoid arthritis rats with bee venom pharmacopuncture, the paw volume of edema of arthritic rats were almost restored to the level of normal group, and behavior tests were very effective. Also the evaluation on the blood serum analysis was remarkable. COX-2, c-fos, and substance-P positive cells in the immunohistological section of dorsal horn region of the spinal cord and subchondral bone of the knee joints were significantly decreased. also the bee venom pharmacopuncture was effective to alleviate their rheumatoid arthritic inflammation cytokine inhibition as regards to the behavior tests and joint histological appearance. Conclusions : Based on the results in this study, bee venom pharmacopuncture with concentrated treatment condition was very effective in low fixed quantity and progressive low increased quantity.

The Biological Effects of Concentrated Growth Factors on the Differentiation and Proliferation of Human Gingival Fibroblasts (Human Gingival Fibroblasts의 분화와 증식에서 CGFs의 생물학적 효과)

  • Park, Sung Il;Bae, Hyun-Sook;Hong, Ki Seok
    • Journal of dental hygiene science
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    • v.12 no.6
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    • pp.689-695
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    • 2012
  • The aim of this study was to elucidate the effects of concentrated growth factors (CGFs) on human gingival fibroblasts in vitro. Blood was collected from three male volunteers (average age 27 years). CGFs were prepared using standard protocols. The CGF exudates were collected at the following culture time points: 1, 7, 14, and 21 days. The levels of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor ${\beta}1$ (TGF-${\beta}1$) in CGFs were quantified. The CGF exudates were then used to culture human gingival fibroblasts. The biologic characteristics of these fibroblasts were analyzed in vitro for 21 days. Platelet-rich plasma released the highest amounts of TGF-${\beta}1$ and PDGF-BB on the first day. The level of TGF-${\beta}1$ had decreased slightly by day 7, although the difference compared to levels at day 1 was not statistically significant. However, by days 14 and 21, levels of TGF-${\beta}1$ had dropped significantly compared to day 1 levels. The levels of PDGF-BB at days 7, 14, and 21 did not differ significantly from that measured on day 1. CGFs maintained the release of autologous growth factors for a reasonable period of time (7 days for TGF-${\beta}1$ and 21 days for PDGF-BB). Gingival fibroblasts treated with CGF exudates collected at day 14 reached peak viability and synthesized type I collagen. Furthermore, the CGF exudates exerted positive effects on the proliferation and differentiation of these cells at days 1, 7, 14, and 21. The findings of this study suggest that treatment with CGFs represents a promising method of enhancing mucosal healing following surgical procedures.