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Immobilization of Bacillus sp. Strains, Catalase Producing Bacteria and Their Hydrogen Peroxide Removal Characteristics (카탈라제를 생산하는 고초균 (Bacillus sp.)의 고정화 및 과산화수소 분해 특성)

  • Han, Kyung-Ah;Jang, Yun-Hee;Rhee, Jong-Il
    • KSBB Journal
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    • v.25 no.6
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    • pp.520-526
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    • 2010
  • In this work we have investigated the production of catalase from Bacillus sp. strains, which were screened and identified from soil. These strains were cultivated in shaking flasks with tryptic soy broth (TSB) at $30^{\circ}C$ and 200 rpm. Effects of the temperature and pH on the stability of the native catalase and whole cell viability were studied in the temperature range of $25-60^{\circ}C$ and the pH range of 7-13. Korean natural zeolite was added to culture medium and mixed with microorganisms for 24 hours. The native catalase maintained its activity over $50^{\circ}C$. The enzyme acitiviy of the catalase from Bacillus flexus BKBChE-3 was highest among the Bacillus sp. strains studied. Bacillus flexus BKBChE-3 and immobilized Bacillus cells have survived under extreme conditions of over $50^{\circ}C$ and pH 12. 60 mL of 10.5 mM $H_2O_2$ solution were entirely removed within 1 hour with catalase produced from Bacillus sp. on the flask. When Bacillus cells were immobilized on Korean natural zeolite, colony forming unit of Bacillus flexus BKBChE-3 was increased and high efficiency of hydrogen peroxide removal was observed.

Effect of $Panax$ $ginseng$ C.A. Meyer Extract (Ginseol K-b1) on UVB-induced Skin Damage in Hairless Mice (Hairless Mice에서 UVB로 유도된 피부손상에 인삼추출물(Ginseol K-b1)이 미치는 영향)

  • Chang, Jeong-Hwa;Hwang, Se-Hee;Lee, Eun-Ju;Ohto, Nobuaki;Lee, Jin-Hee
    • Journal of the East Asian Society of Dietary Life
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    • v.22 no.2
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    • pp.224-230
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    • 2012
  • To investigate the effect of Panax ginseng C.A. Meyer extract (Ginseol K-b1), on skin functionality, we evaluated skin appearance and properties, such as wrinkle formation, skin moisture content, and skin elasticity in the skin of hairless mice damaged by UV irradiation. In addition, the effect of Ginseol K-b1 on collagen synthesis in human dermal fibroblasts was investigated. Female hairless mice were orally administered Ginseol K-b1 for 10 weeks with UV irradiation. Wrinkle formation in the Ginseol K-b1-treated group was significantly suppressed compared to the UV-irradiated group. Skin properties, including skin moisture content and elasticity, of the Ginseol K-b1-treated group were better than those of the control group. In the human fibroblast cells, Ginseol K-b1 treatment enhanced cell proliferation and significantly stimulated collagen synthesis. These results suggest that Ginseol K-b1 is a potent ingredient with anti-aging effects.

The role of ginsenoside Rb1, a potential natural glutathione reductase agonist, in preventing oxidative stress-induced apoptosis of H9C2 cells

  • Fan, Hui-Jie;Tan, Zhang-Bin;Wu, Yu-Ting;Feng, Xiao-Reng;Bi, Yi-Ming;Xie, Ling-Peng;Zhang, Wen-Tong;Ming, Zhi;Liu, Bin;Zhou, Ying-Chun
    • Journal of Ginseng Research
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    • v.44 no.2
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    • pp.258-266
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    • 2020
  • Background: Oxidative stress-induced cardiomyocytes apoptosis is a key pathological process in ischemic heart disease. Glutathione reductase (GR) reduces glutathione disulfide to glutathione (GSH) to alleviate oxidative stress. Ginsenoside Rb1 (GRb1) prevents the apoptosis of cardiomyocytes; however, the role of GR in this process is unclear. Therefore, the effects of GRb1 on GR were investigated in this study. Methods: The antiapoptotic effects of GRb1 were evaluated in H9C2 cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, annexin V/propidium iodide staining, and Western blotting. The antioxidative effects were measured by a reactive oxygen species assay, and GSH levels and GR activity were examined in the presence and absence of the GR inhibitor 1,3-bis-(2-chloroethyl)-1-nitrosourea. Molecular docking and molecular dynamics simulations were used to investigate the binding of GRb1 to GR. The direct influence of GRb1 on GR was confirmed by recombinant human GR protein. Results: GRb1 pretreatment caused dose-dependent inhibition of tert-butyl hydroperoxide-induced cell apoptosis, at a level comparable to that of the positive control N-acetyl-L-cysteine. The binding energy between GRb1 and GR was positive (-6.426 kcal/mol), and the binding was stable. GRb1 significantl reduced reactive oxygen species production and increased GSH level and GR activity without altering GR protein expression in H9C2 cells. Moreover, GRb1 enhanced the recombinant human GR protein activity in vitro, with a half-maximal effective concentration of ≈2.317 μM. Conversely, 1,3-bis-(2-chloroethyl)-1-nitrosourea co-treatment significantly abolished the GRb1's apoptotic and antioxidative effects of GRb1 in H9C2 cells. Conclusion: GRb1 is a potential natural GR agonist that protects against oxidative stress-induced apoptosis of H9C2 cells.

Antiinflammatory Effects of New Chemical Compounds, HS-1580 Series (HS-1580, HS-1581, HS-1582) (신화학물질 HS-1580 유도체(HS-1580 HS-1581 HS-1582)의 항염증 효과)

  • Kim, Ji-Young;Kim, Kyun-Ha;Suh, Hong-Suk;Choi, Won-Chul
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1181-1187
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    • 2006
  • HS-1580 series (HS-1580, HS-1581, HS-1582) can produce anti-imflammatory effects were synthesized from the marine algae extraction in 2,3,6-tribromo-4,5-dihydroxy benzyl methyl ether (TDB). Raw 264.7 cells were pre-treated with $1{\mu}g/{\mu}l$ lipopolysaccharide (LPS) and later treated with HS-1580 series. These cells of inflammatory mediators were tested as well. Nitric oxide (NO) is related to autoimmune disease and is produced by inducible NOS (iNOS). When treated with HS-1580 series, the product of NO will reduce in a dose-dependent manner. HS-1580 series significantly inhibit the iNOS protein expression. Cyclooxygenase (COX) involves with the various physiologic events and catalyzes in prostaglandin. HS-1580 series also inhibit the COX-2 protein expression as well as pro-inflammatory cytokines production such as tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})\;and\;interluekin-1{\alpha]\;(IL-1{\beta})$. These upcoming results suggest that HS-1580 series have anti-inflammatory efforts in Raw 264.7 cells by inhibiting such as iNOS, COX-2, $TNF-{\alpha}\;and\;IL-1{\beta}$ as inflammatory mediators.

The Effect of NO Donor on Contraction, Cytosolic $Ca^{2+}$ Level and Ionic Currents in Guinea-pig Ileal Smooth Muscle

  • Kwon, Seong-Chun;Park, Ki-Young;Ahn, Duck-Sun;Lee, Young-Ho;Kang, Bok-Soon
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.1
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    • pp.33-40
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    • 2000
  • This study was designed to clarify the mechanism of the inhibitory action of a nitric oxide (NO) donor, 3-morpholino-sydnonimine (SIN-1), on contraction, cytosolic $Ca^{2+}$ level $([Ca^{2+}]_i)$ and ionic currents in guinea-pig ileum. SIN-1 $(0.01{\sim}100\;{\mu}M)$ inhibited 25 mM KCl- or histamine $(10\;{\mu}M)-induced$ contraction in a concentration-dependent manner. SIN-1 reduced both the 25 mM KCl- and the histamine-stimulated increases in muscle tension in parallel with decreased $[Ca^{2+}]_i.$ Using the patch clamp technique with a holding potential of -60 mV, SIN-1 $(10\;{\mu}M)$ decreased peak Ba currents $(I_{Ba})$ by $30.9{\pm}5.4%$ (n=6) when voltage was stepped from -60 mV to +10 mV and this effect was blocked by ODQ $(1\;{\mu}M),$ a soluble guanylyl cyclase inhibitor. Cu/Zn SOD (100 U/ml), the free radical scavenger, had little effect on basal $I_{Ba},$ and SIN-1 $(10\;{\mu}M)$ inhibited peak $I_{Ba}$ by $32.4{\pm}5.8%$ (n=5) in the presence of Cu/Zn SOD. In a cell clamped at a holding-potential of -40 mV, application of $10\;{\mu}M$ histamine induced an inward current. The histamine-induced inward current was markedly and reversibly inhibited by $10\;{\mu}M$ SIN-1, and this effect was abolished by ODQ $(1\;{\mu}M).$ In addition, SIN-1 markedly increased the depolarization-activated outward $K^+$ currents in the all potential ranges. We concluded that SIN-1 inhibits smooth muscle contraction mainly by decreasing $[Ca^{2+}]_i$ resulted from the inhibition of L-type $Ca^{2+}$ channels and the inhibition of nonselective cation currents and/or by the activation of $K^+$ currents via a cGMP-dependent pathway.

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Saprolegnia parasitica Isolated from Rainbow Trout in Korea: Characterization, Anti-Saprolegnia Activity and Host Pathogen Interaction in Zebrafish Disease Model

  • Shin, Sangyeop;Kulatunga, D.C.M.;Dananjaya, S.H.S.;Nikapitiya, Chamilani;Lee, Jehee;De Zoysa, Mahanama
    • Mycobiology
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    • v.45 no.4
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    • pp.297-311
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    • 2017
  • Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

The Growth of Osteoblasts according to the Pretreatment of HA Scaffold Surface (하이드록시아파타이트 지지체의 표면 처리 방법에 따른 골아세포의 성장)

  • Park, Byung-Chan;Kim, Yong-Ha;Kim, Tae-Gon;LeeYoun-Jung, Jun-Ho;Sik-Young, Kim;Choi, Sik-Young
    • Archives of Plastic Surgery
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    • v.37 no.4
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    • pp.340-345
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    • 2010
  • Purpose: Recently, bioceramics have become popular as a substitute graft material for reconstruction of bony defect after trauma or tumor surgery. Among the bioceramic materials, hydroxyapatite (HA) is favored due to its biocompatibility. HA scaffold is composed of the interconnected reticular framework, macropores and micropores. Macropores play an important role in cell migration, nutrients supply and vascular ingrowth. On the other hand, a number of micropores less than $10{\mu}m$ form an irregular surface on HA scaffolds, which prevents the osteoblast from adhering and proliferating on the surface of HA scaffold. Methods: In this study, three different groups were designed for comparison. In the first group (group A), conventional method was used, in which HA pellet was applied without surface pretreatment. The second group (group B) was given a HA pellet that has been coated with crystalline HA solution prior to application. In the third group (group C), the same method was used as the second group, where the pretreated HA pellet was heated ($1250^{\circ}C$, 1 hour) before application. Osteoblast-like cells ($2{\times}10^4$/mL) were scattered onto every pellet, then they were incubated in 5% $CO_2$ incubator at $37^{\circ}C$ for twelve days. During the first three days, osteoblast cells were counted using the hemocytometer daily. ALP activity was measured on the 3, 6, 9 and 12 culture days using the spectrophotometer. Results: Under SEM, group A showed a surface with numerous micropores, and group B revealed more rough crystal surface. Group C revealed a fused crystal appearance and flattened smooth surface. In proliferation and ALP activity of osteoblast cells, group C showed better results compared to group B. Group A which lacks pretreatment of the surface showed less osteoblast proliferation and ALP activity than group C, but showed better results than group B. Conclusion: We found that crystallized HA with heat treatment method enhances the osteoblasts proliferation and differentiation on the surface of HA pellets.

Chemical and Morphological Change and Discoloration of Cedar Wood Stored Indoor (실내 보관 삼나무 목재의 재색 및 화학적·현미경적 변화 특성)

  • Lee, Kwang Ho;Cha, Mi Young;Chung, Woo Yang;Bae, Hyeun-Jong;Kim, Yoon Soo
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.6
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    • pp.566-577
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    • 2009
  • The modification of wood color occurs rapidly during the service period at indoor. It is crucial to investigate the characteristics of color change, chemical and microscopical modification of wood at indoor. Wood products made of Japanese cedar at different years were used for this work. The tests were performed in order to evaluate the degree of color change of wood surface, breakpoint of brightness from surface to inside of wood, chemical analysis with FT-IR, and microscopical characteristics using the LM and TEM. Surface color of cedar wood stored indoor were rapidly changed at early stage, particularly ${\Delta}a$ (yellow), and ${\Delta}b$ (red) values were steeply decreased for one year old indoor wood, ${\Delta}L$ (white) value was dropped until 5 years old indoor wood compared with control sample. Decrease of peaks related to polysaccharide and lignin was noticed, especially, lignin was severely degraded. Although degradation of cell wall limited only to surface layers of indoor wood, degradation pattern of indoor wood showed similar degradation pattern to natural weathering of wood during outdoor weathering or wood behavior under artificial UV irradiation.

Ingestion rate and grazing impact by the mixotrophic ciliate Mesodinium rubrum on natural populations of marine heterotrophic bacteria in the coastal waters of Korea

  • Seong, Kyeong Ah;Myung, Geumog;Jeong, Hae Jin;Yih, Wonho;Kim, Hyung Seop;Jo, Hyun Jung;Park, Jae Yeon;Yoo, Yeong Du
    • ALGAE
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    • v.32 no.1
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    • pp.47-55
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    • 2017
  • We explored feeding by the mixotrophic ciliate Mesodinium rubrum, heterotrophic nanoflagellates (HNFs), and small ciliates (<$30{\mu}m$ in cell length) on natural populations of heterotrophic bacteria in Masan Bay, Keum River Estuary, and in the coastal waters of the Saemankeum area, Korea when M. rubrum red tides occurred. We also measured ingestion rates of M. rubrum on cultured heterotrophic bacteria as a function of bacterial concentration in the laboratory. The ingestion rates of M. rubrum on natural populations of heterotrophic bacteria (2.3-16.8 bacteria $grazer^{-1}h^{-1}$) were comparable to or lower than those of co-occurring HNFs (10.7-41.7 bacteria $grazer^{-1}h^{-1}$), but much lower than those of co-occurring small ciliates (76.0-462.2 bacteria $grazer^{-1}h^{-1}$). However, the maximum grazing coefficient of M. rubrum ($0.245d^{-1}$) on natural populations of heterotrophic bacteria was much higher than that of small ciliates ($0.089d^{-1}}$), and slightly higher than that of HNFs ($0.204d^{-1}$). With increasing bacterial concentrations, ingestion rates of M. rubrum on cultured heterotrophic bacteria continuously increased, but became saturated at higher prey concentrations over $1-5{\times}10^6cells\;mL^{-1}$. The maximum ingestion rate of M. rubrum on cultured heterotrophic bacteria was 34.4 bacteria $grazer^{-1}h^{-1}$. Based on the present study, it is suggested that M. rubrum may be an important grazer of heterotrophic bacteria and sometimes have considerable grazing impact on natural populations of heterotrophic bacteria.

Effect of Si-Wu-Tang and Si-Jun-Zi-Tang on the Survival of Jejunal Crypt Cells and Hematopoietic Cells in Irradiated Mice (방사선조사 마우스에서 소장움세포 및 조혈세포 생존에 미치는 사물탕 및 사군자탕의 영향)

  • Kim, Sung-Ho;Oh, Heon;Lee, Song-Eun;Jo, Sung-Kee;Byun, Myung-Woo
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.888-894
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    • 1998
  • In order to investigate the radioprotective effect of Si-Wu-Tang (Korean name: Sa-Mul-Tang), a kind of traditional Oriental medicine as a blood-building decoction (Oriental medical concept: Bu-Xie), and Si-Jun-Zi-Tang (Korean name: Sa-Gun-Ja-Tang), one of the widely used Oriental herbal medicines as an energy tonic (Chinese medical concept: Bu-Qi). the jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells were observed in irradiated mice. Jejunal crypts were protected by Si-Wu-Tang pretreated both per os (2 mg/mL of drinking water for 7 days, p<0.05) and intraperitoneally (1 mg/head, single injection at 24 hours before irradiation). Si-Wu-Tang adminstration before irradiation(1 mg/head, single injection at 24 hours before irradiation) resulted in an increase of the formation of endogenous spleen colony (p<0.005). The frequency of radiation-induced apoptosis in intestinal crypt cells was also reduced by pretreatment of Si-Wu-Tang (p<0.01). However, the radioprotective effect of Si-Jun-Zi-Tang was not as significant as that of Si-Wu-Tang. These results suggest that Si-Wu-Tang may be a useful radioprotective food, especially since it is a relatively nontoxic natural product.

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