Background: Focal adhesions (FAs) is the most important process in the first step of osseointegration between preosteoblasts and titanium (Ti). FAs improvement and pre-osteoblasts cell proliferation leads to successful Ti-based dental implants. This study aimed to confirm the applicability of rosmarinic acid (RA) as a functional substance for improving FAs and cell proliferation of MC3T3-E1 preosteoblasts on Ti surfaces during the first stage of osseointegration for successful Ti-based dental implants. Methods: We used MC3T3-E1 preosteoblasts on Ti discs incubated in a medium supplemented with or without 14 ㎍/ml to decipher the effects of RA on FAs and cell proliferation. FAs and proliferation of MC3T3-E1 cells on Ti discs were assessed via MTT assay. Actin-labeled cells and paxillin contacts were observed and imaged by fluorescent microscopy, and the associated signaling pathways were revealed through western blot analysis. Results: In RA-treated MC3T3-E1 cells on Ti discs, FAs between MC3T3-E1 preosteoblasts and Ti surfaces and the expression of focal adhesion kinase (FAK), phosphorylated FAK and paxillin proteins and filamentous-actin formation increased. RA increased the proliferation of MC3T3-E1 preosteoblasts on the Ti surface as well as the expression of Grab2, Ras, pERK1/2, and ERK1/2. In addition, the expression of secretory leukocyte protease inhibitor and thymosin b4, known as nanomolecules that enhance the interaction between implanted Ti materials and preosteoblasts in the RA-treated MC3T3-E1 preosteoblasts, increased. RA not only increased the FAs of MC3T3-E1 preosteoblasts on the Ti surface through the FAK/Paxillin signaling pathway, but also increased cell proliferation and mitosis through the FAK/Grab2/Ras/ERK1/2 signaling pathway. Conclusion: RA can be applied as an effective functional substrate to improve the FAs and proliferation of MC3T3-E1 preosteoblats on Ti surfaces, which are essential in the first step of osseointegration between implanted Ti and bone tissue for the clinical success of Ti based dental implants.
Yoon, Ho Il;Lee, Chang-Hoon;Lee, Hee-Seok;Lee, Choon-Taek;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yoo, Chul-Gyu
Tuberculosis and Respiratory Diseases
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v.54
no.4
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pp.449-458
/
2003
Background : Cyclosporin A(CsA) and tacrolimus(FK506) have been widely used as immunosuppressants. The effects of CsA, or FK506, on the $I{\kappa}B/NF-{\kappa}B$ pathway have been shown to vary according to the cell type. However, their effects on the $I{\kappa}B/NF-{\kappa}B$ pathway have not been reported in bronchial epithelial cells. In this study, the effects of CsA and FK506 on the $I{\kappa}B/NF-{\kappa}B$ pathway in bronchial epithelial cells, monocytes, lymphocytes and alveolar macrophages were evaluated. The relationship between their effects on the $I{\kappa}B/NF-{\kappa}B$ pathway and $I{\kappa}B$ kinase(IKK) activity was also investigated. Methods : BEAS-2B and A549 cells, pulmonary alveolar macrophages, peripheral blood monocytes and lymphocytes were used. The cells were pre-treated with CsA, or FK506, for various time periods, followed by stimulation with TNF-${\alpha}$, LPS or IL-$1{\beta}$. The $I{\kappa}B{\alpha}$ expressions were assayed by Western blot analyses. The IKK activity was evaluated by an in vitro immune complex kinase assay, using GST-$I{\kappa}B{\alpha}$ as the substrate. Results : Neither CsA nor FK506 affected the level of $I{\kappa}B{\alpha}$ expression in any of the cell types used in this study. CsA pre-treatment inhibited the TNF ${\alpha}$-induced $I{\kappa}B{\alpha}$ degradation in bronchial epithelial cells. In contrast, the TNF ${\alpha}$-induced $I{\kappa}B{\alpha}$ degradation was not affected by FK506 pre-treatment. However, FK506 suppressed the cytokine-induced $I{\kappa}B{\alpha}$ degradation in the pulmonary alveolar macrophages, peripheral blood monocytes and lymphocytes. The inhibitory effect of CsA, or FK506, on $I{\kappa}B{\alpha}$ degradation was not related to IKK. Conclusions : CsA and FK506 suppressed the $I{\kappa}B{\alpha}$ degradation in bronchial epithelial cells, monocytes, lymphocytes and alveolar macrophages, so this may not be mediated through IKK.
An experimental comparative study was done to determine the protective effects of three preservation solutions on isolated rabbit heart-lung bloc during acute ischemia and reperfusion of the lung during lung transplantation. Thirty Isolated rabbit heart-lung blocs were divided into 3 groups , group I(n:9) was preserved with Hartmann's solution, group II(n: 10) with modified University of Wisconsin solution, and group III(n: 1 1) with Kosin solution. The isolated heart-lung blocs were washed with Hartmann's so ution. Aftar infusion of each preservation solution into pulmonary artery, the heart-lung bloc was stored at 4'c cold preservation solution for each group for 4 hours and .then the heart-lung blocs were reventilated and reperfused. The changes of weight of heart-lung blocs, airway pressure, percent change of PCO2, level of lactate and adenosine deaminase(ADA) and microscopic structure of the lung parenchyme were evaluated. Results were as follows 1. A change of weight of the heart lung bloc after reperfusion was lowest in group 111(p< .05) 2. The airway pressure increased after reperfusion in group I but decreased in group II, and II. Especially in group II, post-reperfusion airway pressure returned to level lower than that of en-bloc resection. 3. Pulmonary artery pressure during reperfusion after 4 hour preservation was lowest in group III, and pulmonary artery pressure in group II was higher than in group I(P> 0.1). 4. The level of lactate and ADA in the lung tissue were higher in group III than in group I and II(P< .05) 5. The percent change of PCO2 in perfusate was slightly higher in group III than group I and II. 6. Microscopic changes in lung tissue after reperfusion were diffuse pulmonary edema, expansion of inter- stitial tissue, focal aggregation of erythrocytes, and basement membrane abnormalities, but no differences were found among the three groups. In conclusion, the protective effects of modified University of Wisconsin solution and Kosin solution were slightly superior to Hartmann's solution.
Kim, Y.I.;Jun, S.H.;Yang, S.Y.;Huh, J.W;Kwak, W.S.
Journal of Animal Science and Technology
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v.49
no.5
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pp.667-676
/
2007
This study was conducted to determine effects of cellulolytic microbes inoculation to sawdust-based spent mushroom substrate(SMS) during deepstacking on fermentation parameters, total microbial counts and cellulolytic enzyme activity and to on SMS nutrients utilization by sheep. For sheep metabolism trials, six sheep(ram, average 54.8kg) were fed a Control diet(70% concentrates, 15% rice straw and 15% SMS with no microbial treatment on a dry basis) and a Treatment diet(the same diet including SMS with a microbial treatment) for 2 trials. Spent mushroom substrates with or without a microbial(4 strains including 1 strain of Enterobacter ludwigii, 1 strain of Bacillus cereus and 2 strains of Bacillus subtillis) treatment (1% of SMS on wet basis) were deepstacked for 7 days. The internal temperatures in 1.2 M/T of SMS deepstacks reached to 50±5℃ within 7 days of storage. Total microbial counts remarkably decreased (P<0.05) with a deepstacking process and were not affected(P>0.05) by the microbial treatment. For fibrolytic enzyme activity, CMCase and xylanase activities were decreased(P<0.05) by a deepstacking process. After deepstacking, the microbial treatment showed about 2.5-times higher(P<0.05) for CMCase activity and about 4-times higher(P<0.05) for xylanase activity than those of the Control. Activities of ligninolytic enzymes such as laccase and MnP were not affected by the microbial treatment. The sheep fed the microbially treated SMS diet had a tendency of greater total tract digestibilities of ash(P=0.051), NFE (P=0.071), hemicellulose(P=0.087) and NDF(P=0.096) than those fed the untreated SMS diet. Nitrogen balance of sheep was not affected(P>0.05) by feeding of microbially treated SMS. Accordingly, these results indicate that cellulolytic microbes inoculation during deepstacking of SMS may improve the bio- utilization of SMS by sheep.
This study was conducted to investigate effects of terpenoids-rich plant extracts (TRPE) on the in vitro ruminal fermentation characteristics and methane production. The ruminal fluid was collected from a cannulated Hanwoo cow fed concentrate and timothy in the ratio of 6 to 4. The TRPE as Mint (Mentha arvensis var. piperascens), Pine (Pinus densiflora), Japan cedar (Cryptomeria japonica), Sichuan pepper (Zanthoxylum piperitum), Hinoki cypress (Chamaecyparis obtuse) and Japanese black pine (Pinus thunbergii) were used in this study. The 15 mL of mixture, contains McDougall buffer and rumen fluid in the ratio of 2 to 1. The mixture was dispensed anaerobically 50 mL serum bottles and it is contained 0.3 g timothy substrate and 5% TRPE. The bottles were incubated at $39^{\circ}C$ for 3, 6, 9, 12, 24, 48 and 72 hours. The pH value decrease by increased incubation times and the pH values at all times were significantly (p<0.05) higher in treatments than in control. The digestibility of dry matter at 3 hours was significantly (p<0.05) higher in mint treatment than in control. Productions of total gas and carbon dioxide at before 12 hours was significantly lower (p<0.05) in treatments than in control. The methane production at 24 hours was significantly (p<0.05) lower in treatments than in control. The concentrations of acetic acid and propionic acid at 24 hours were significantly higher (p<0.05) in mint and pine treatments than in control. In conclusion, the terpenoid-rich plant extracts were shown to decreased methane emission and without adversely affected ruminal fermentation. Therefore, the terpenoid-rich plant extracts as mint and pine were shown to decreased methane production and it has potential possibility for ruminal fermentations.
Kim, Jae-Young;Kim, Bong-Kyu;Yi, Yong-Sub;Kang, Chang-Soo;Ahn, Joong-Hoon;Lim, Yoong-Ho
Microbiology and Biotechnology Letters
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v.37
no.2
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pp.99-104
/
2009
The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.
The primary objective of this study was to examine the toxic effects of PCP on activated sludge and to analyze its metabolic responses while treating wastewater containing pentachlorophenol (PCP) in a sequencing batch reactor (SBR) system operating under different control strategies. This study was conducted in two phases 1 and 2 (8-hr and 12-hr cycles). Each phase was operated with two control strategies I and II. Strategy I (reactor 1) involved rapid addition (5 minutes to complete) of substrate to the reactor with continuous mixing but no aeration for 2 hours. Strategy ll (reactor 2) involved adding the feed continuously during the first 2 hours of the cycle when the system was mixed but not aerated. During both phases each reactor was operated at a sludge age of 15 days. The synthetic wastewater was used as a feed. The COD of the feed solution was about 380 mg/l. After the reference response for both reactors was established, the steady state response of each system was established for PCP feed concentrations of 0.1 mg/l, 1.0 mg/l, and 5.0 mg/l in SBR systems operating on both 8-hr and 12-hr cycles. Soluble COD removal was not inhibited at any feed PCP concentrations used. At 5.0 mg/l fined PCP concentration and in SBR systems operating on phase 2, the concentrations of MLVSS were decreased; selective pressure on the mixed biomass might be increased, narrowing the range of possible ecological responses; the settleability of activated sludge was poor; the SOURS were increased, showing that the systems were shocked. Nitrification was made to some extent at all concentrations of feed PCP in SBR systems operating on phase 2 whereas in SBR systems operating on phase 1 little nitrification was observed. Then, nitrification will be delayed as much as soluble COD removal is retarded due to PCP inhibition effects. Enhanced biological phosphorus removal occurring in the system operating with control strategy I during phase 1 of this work and in the presence of low concentrations of PCP was unreliable and might cease at anytime, whereas enhanced biological phosphorus removal occurring in the system operating with either control strategy I or II during phase 2 of this work and in the Presence of feed PCP concentrations up to 1.0 mg/l was reliable. When, however, such processes were exposed to 5.0 mg/l PCP dose, enhanced phosphorus removal ceased and never returned.
Kim, Ki-Deog;Lee, Eung-Ho;Kim, Won-Bae;Lee, Jun-Gu;Yoo, Dong-Lim;Kwon, Young-Seok;Lee, Jong-Nam;Jang, Suk-Woo;Hong, Soon-Choon
Journal of Bio-Environment Control
/
v.20
no.2
/
pp.116-122
/
2011
This study was carried out to investigate the effects of several cooling methods such as water hose cooling, mist, fog and control on growth and microclimate, and to develop a simple nutriculture bed for production of fresh leaves of narrowhead goldenaray 'Ligularia stenocephala'. When the root-zone was cooled with 240 L/hr flow rate of $13^{\circ}C$ ground water using water hose, the temperature was lowered approximately by 2 to $3^{\circ}C$ than that of control. The growth of narrowhead goldenaray were favorable in the water hose cooling compared with the other cooling methods. Nutrient culture system having part cooling effect around plant canopy was developed. The system was composed of 15 cm diameter of water hose on side wall of beds, cooling hose, and expanded rice hull media as organic substrate. When cool water which the temperature changed in the range of 14 to $22^{\circ}C$ diurnally with 240 L/hr of flow rate through water hose, the air temperature around canopy and root-zone temperature were dropped by $0.5^{\circ}C$ and $3^{\circ}C$ compared with that of conventional styrofoam bed, respectively. These results showed that newly devised bed system using water hose was simple and economical for the production of high quality narrowhead goldenaray leaves. This system might be practically used both at summer and winter season for the cultivation of narrow head goldenaray by part cooling or heating around root-zone and plant canopy.
This study was conducted to investigate the effects of fermented spent mushroom substrates (F-SMS) of Flammulina velutipes on growth performance, carcass traits, and economic characteristics of Hanwoo steers. A yeast strain (Saccharomyces sp. UJ14) and Bacillus strain (Bacillus sp. UJ03) isolated from fresh spent mushroom substrates of Flammulina velutipes were used as probiotics to prepare F-SMS. Twenty-four Hanwoo steers (14 months old) were allocated to three dietary treatments via a randomized block design and were slaughtered at 30 months of age. These treatment groups included Control (TMR), T1 (TMR containing 10% of F-SMS) group, and T2 (TMR containing 30% of F-SMS). Body weight gain was not influenced by the experimental diets. DM and TDN intakes in the finishing period were significantly (p < 0.05) greater in group T1 than in other groups. CP intake was significantly (p < 0.05) greater in group T2 than in other groups during the whole experimental period. Among carcass traits, rib-eye area and back fat thickness tended to increase with F-SMS supplementation. The appearance rate (%) of a meat yield more than grade A was the highest in group T1. The net profits increased by 1.2% and 13.3% in groups T1 and T2, respectively. In conclusion, if a proper feeding program (including feed safety) can be ensured, spent mushroom substrates of Flammulina velutipes can prove to be a highly profitable feed source for Hanwoo steers.
We evaluated preventive effects of Suaeda japonica (SJ) and Spergularia marina Griseb (SMG) on the insulin resistance in Otsuka Long-Evans Tokushima Fatty (OLETF) rats. The 10-week old OLETF rats were fed diets containing 3% (w/w) SJ and SMG for 18 weeks. Fasting blood glucose levels in SJ and SMG groups, measured using the oral glucose tolerance test, were lower than that of the control rats. The SMG group showed significantly lower levels of insulin, glycated hemoglobin, triglyceride, and total cholesterol than the control group. In addition, these levels were relatively lower in the SJ group than those in the control rats. The SJ and SMG groups had relatively lower protein levels of nuclear factor-kappa B (NF-${\kappa}B$) p65 in adipose tissue and serine phosphorylated insulin receptor substrate 1 (IRS-1) in skeletal muscle than the control group. These results suggest that SJ and SMG prevent insulin resistance and SMG in particular reduces blood triglyceride and total cholesterol levels.
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