• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.277-288
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• 2003

It has been suggested that increased number and activity of phagocytes in periodontitis lesion results in a high degree of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide, nitric oxide and peroxynitrite. There are few reports on the relationship between ROS and MMPs expressions in gingival fibroblast. We studied to elucidate whether and how ROS, especially nitric oxide affects the MMP expression. Human gingival fibroblasts and HTl080 cells (human fibrosarcoma sell line as reference) were grown in DMEM supplemented with 10 mM HEPES, 50 mg/L gentamicin, and 10% heat inactivated fetal bovine serum with addition of various reactive oxygen species (ROS). Culture media conditioned by cells were examined by gelatin zymography. HT1080 cells expressed proMMP-2 and proMMP-9, but human gingival fibroblasts (HGF) produced only proMMP-2. Hydrogen peroxide upregulated MMP-9 expression in HT1080 cells, whereas in human gingival fibroblast SNP treatment showed marked increase in MMP-2 level compared to other ROS. These results suggest that the effects of ROS on MMPs expressions are cell-type specific. RT-PCR for MMP-2 and TIMP-2 m-RNA were performed using total RNA from cultured cells under the influence various kinase inhibitors. In HT1080 cells, treatment with FPTI III (Ras processing inhibitor) and LY294002 (PI3-kinase inhibitor) resulted in inhibition of MMP-2 and MMP-9 expressions, suggesting that Ras/P13-kinase pathway is important for MMPs expression in HT1080 cells. In gingival fibroblasts, treatment with FPTI III and PDTC (NF-kB inhibitor) showed marked decrease in MMP-2 regardless of the of SNP , suggesting that Ras/NF-kB could be the key pathway for NO-induced MMP-2 expression in gingival fibroblasts. This study showed that ROS, especially nitric oxide, could be the critical mediator of periodontal disease progression through control of MMP-2 expression in gingival fibroblasts possibly via Ras/NF-kB pathway.

• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.1013-1021
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• 1996

There are many important factors in periodontal inflammation. $IL-1{\beta}$, $PGE_2$ and collagenase are predorminantly key factors. These inflammatory mediators induce gingival tissue and alveolar bone destruction. For the prevention and treatment of periodontal disease, it is necessary to inhibit $IL-1{\beta}$, $PGE_2$ production and collagenase activity. Ursodeoxycholic acid(UDCA) has immunomodulatory properties, and there is evidence that some natural extracts show antiinflammatory activity to some degree. The purpose of this study was to assess the inhibitory effect of UDCA and its mixture with natural extracts on $IL-1{\beta}$, $PGE_2$ production and collagenase activity. Accordingly we assessed the effect of UDCA and its mixture combined with some natural extracts on inhibition of $IL-1{\beta}$, $PGE_2$ production and collagenase activity. For the $IL-l{\beta}$ inhibition study, cultured cells were exposed to $25{\mu}g/ml$ LPS. $IL-1{\beta}$ activity was measured by $IL-1{\beta}$ enzyme immunoassay system. Human gingival fibroblasts were prepared and cells (l05/well) were seeded into culture plates. $rhIL-1{\beta}$ was added to induce $PGE_2$. The amount of $PGE_2$ in sample media was measured using enzyme immunoassay system. Crude collagenase was prepared from Porphyromonas gingivalis and collagenolytic activity was determined using a Collageno kit CLN-100. The test inhibitor was added to the assay mixture consisting of 0.1ml of 50mM Tris buffer(pH 7.5) and 0.2ml of substrate solution. UDCA and UDCA combined with natural extracts generally inhibited $IL-1{\beta}$ production. groups above 0.01% UDCA strongly inhibited $IL-l{\beta}$ synthesis. Both groups inhibited $IL-1{\beta}-induced$ synthesis of $PGE_2$. In low concentration, the degree of inhibition was as same as prednisolone. In high concentration, each group was superior to prednisolone. UDCA group and UDCA mixture group exerted a moderate inhibition of collagenolytic enzyme. The present study suggested that UDCA and its mixture with natural extracts could be further investigated as antiinflammatory drug for periodontal disease.

• Herbal Formula Science
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• v.14 no.2
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• pp.76-85
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• 2006

Objectives : We Investigated the effect of Dioscoreae Rhizoma(山藥) on the change of the corticosterone and the rectal temperature(直腸溫渡) of the mice induced by starvation stress(創戰 스트레스). Methods : After administration of Dioscoreae Rhizoma (0.25g/kg, 0.5g, 1.0g/kg, 3g/kg) three times, mice were starved. The corticosterone and rectal temperature were measured after 36.5 hours starvation stress. Results : The plasma cortiosterone levels in the S-2, S-3 and S-4 group were decreased significantly comparing with the control group (P<0.01) after 36.5 hours starvation stress. and rectal temperature was decreased in the control goup comparing with the normal group, but there is no significant change in the Dioscoreae Rhizoma treated group. Conclusion : it might be recognized that Dioscoreae Rhizoma has preventive-effect against starvation stress and also it might be needed further study in various viewpoints. Objectives : This study was disegned to elucidate the short term effect of Rossa rugosae Radix on proliferation. differentiation and maturation of 3T3-L1 Preadipocyte. Methods: 3T3-L1 preadipocytes obtained from Korean Cell Line Bank were cultured in a D ulbecco’ s modified eagle medium(MEM) culture solution containing 10% fetal bovine serum(FBS) and various concentrations of aqueous extract of Rossa rugosae Radix.. The short term effect of the extract of Rossa rugosae Radix on proliferation. differentiation and maturation of 3T3-L1 preadipocytes were investigate after treatment for 24 hours by measuring MTT. Oil Red 0 and latate dehydrogenase activity.. Results: The Rossa rugosae Radix extract inhibited significantly the proliferation of 3T3-L1 preadipocytes and tended to increase latate dehydrogenase activity in the media of differentiated 3T3-L1 preadipocytes & matured 3T3-L1 preadipocytes. the extract also inhibit the lipid accumulation of differentiated 3T3-L1 preadipocytes & matuered 3T3-L1 preadipocytes. Conclusions: These results demonstrated that the Rossa rugosae Radjx extract inhibited the proliferation. differentiation and maturation of 3T3-L1 preadipocytes. suggesting that Rossa rugosae Radix has anti-obesity effect: however further in vivo study is needed to demonstrate its pharmacological effects.

• Journal of Plant Biotechnology
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• v.38 no.4
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• pp.251-257
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• 2011

Rice is the most important crop as a model plant for functional genomics of monocotyledons. Rice is usually transformed using Agrobacterium tumefaciens. However, the transformation efficiency using previous method is still low. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the inoculation and co-cultivation step. We directly inoculated Agrobacterium containing a CIPK15 gene under the control of CaMV 35S promoter and NOS terminator in the pCAM1300 vector into the pre-soaked seeds in N6D media for 24 hours. After 7 days of culture at $25^{\circ}C$, calli were formed on seeds cultured on the co-cultivation medium containing an antioxidant compound (1 mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (3 mg/L silver nitrate). We obtained 35 and 22 transgenic plants in rice cultivars, Gopumbyeo and Ilpumbyeo, with increase of transformation efficiency by 30.4% and 22.6%, respectively compared to the general transformation method. The new method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

• Journal of Plant Biotechnology
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• v.45 no.3
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• pp.266-272
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• 2018

A Polygonatum stenophyllum Maxim. is an important endangered plant belonging to the family Liliaceae. A method was developed for the rapid micropropagation of P. stenophyllum through plant regeneration from rhizome (1-year, 3-years, and 5-years) explant-derived calli. The rhizome segments were cultured in Murashige and Skoog (MS) medium supplemented with varying concentrations of 2,4-D (0, 0.5, 1.0, $1.5mg{\cdot}L^{-1}$) for callus induction. In media supplemented with $0.5mg{\cdot}L^{-1}$ of 2,4-D, 87% of 3-years rhizome produced callus. Subsequently, the callus was transferred to 1/2MS medium supplemented with various concentrations of IAA, IBA, NAA, and 2,4-D (0, 0.1, 0.5 and $1.0mg{\cdot}L^{-1}$) for adventitious shoot formation. The highest percentage of adventitious shoot induction (57%) was observed in 1/2MS medium containing $0.5mg{\cdot}L^{-1}$ of NAA. Elongation of the adventitious shoot was achieved in 1/2MS medium supplemented with $0.1mg{\cdot}L^{-1}$ of BA. Rooting was achieved in 1/2MS medium without any hormones. It is hypothesized that the stated in vitro propagation protocol will be useful for conservation and mass propagation of the endangered Polygonatum stenophyllum Maxim. for bioresources.

• Research in Plant Disease
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• v.13 no.3
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• pp.191-196
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• 2007

An isolate of Ampelomyces quisqualis 94013(AQ94013) was selected as an effective agent for biological control against cucumber powdery mildew. In order to develop mass production technique, six cereal media made with barley, rice, mille and brown rice, sorghumand rice seed were tested. Among them, barley medium was the best for the growth and conidial production of AQ94013. Optimum temperature for the mass production of AQ94013 was $25^{\circ}C$ and followed by $20^{\circ}C$ and $30^{\circ}C$. Light radiation inhibited conidial production of AQ94013 since number of conidia formed on barely medium under continuous light or 12 hrs alternative light were much less than cultured in darkness. Tthe conidia produced on the medium at $30^{\circ}C$ maintained the parasitic ability to Sphaerotheca fusca for 30 days. A culture method of AQ94013 in barley liquid medium with adding barely power(40 g/l) in darkness for five days at $25^{\circ}C$using a 30 l-fermenter was very effective for mass production of conidia.

• Journal of Life Science
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• v.15 no.1 s.68
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• pp.118-123
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• 2005

For the expression in Saccharomyces cerevisiae, Bacillus stearothermophilus cyclodextrin glucano­transferase gene (cgtS) in pCGTS (4.8 kb) was subcloned into the surface expression vector, pYD1 (GALl promoter). The constructed plasmid, pYDCGT (7.2 kb) was introduced into S. cerevisiae EBY100 cells, and then yeast transformants were selected on the synthetic defined media lacking tryptophan. The formation of cyclodextrin (CD) was confirmed with active staining of culture broth of transformant grown on starch medium. Enzymatic reaction products with respect to the culture time and the reaction time were examined by TLC analysis. The results indicated that the enzyme activity was exhibited after 12 h cultivation and CD was produced after 10min of enzymatic reaction. When the surface-engineered yeast cells were cultured on galactose medium, maximum activities of CGTase were about 21.3 unit/l and 16.5 unit/l at $25^{\circ}C\;and\;30^{\circ}C$, respectively. The plasmids stability showed about $80\%\;even\;at\;25^{\circ}C\;and\;30^{\circ}C$.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.1
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• pp.45-53
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• 2009

Objectives: To determinate the optimal culture condition to maintain lifespan in human sperm, we evaluated the effect of different temperature on sperm motility and viability up to 5 days in normal specimens. Methods: Ejaculated semen samples with normal semen parameters were gently washed in HEPES buffered Tyrod's-Albumin-Lactate-Pyruvate (HTALP) media. Each 5 ml of HTALP + 0.3% albumin with $1{\times}10^6$ sperm/ml was incubated for 5 days in $37^{\circ}C$, $22^{\circ}C$, and $4^{\circ}C$. The sperm motility and kinematics were analyzed using computer assisted sperm analysis (CASA), membrane integrity was assessed by hypoosmotic swelling test (HOST), and capacitation status was evaluated by chlorotetracycline (CTC) fluorescence pattern. Each parameter was measured on day 1, 3, and 5, respectively. Results: The motility, viability and live/uncapacitated pattern were demonstrated significantly in temperature- and time-dependent difference (p<0.05). While the sperm cultured for 1 day in each temperature was not significantly different, the sperm cell kept in $22^{\circ}C$ after 3 days were preserved sperm motility, viability, membrane integrity, and F pattern better than in other culture temperatures. Conclusions: HTALP can be used a basic medium for culture and longevity preservation, and sperm cell kept at $22^{\circ}C$ is beneficial for assisted reproductive techniques.

• Asian Journal of Turfgrass Science
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• v.25 no.2
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• pp.184-189
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• 2011

To optimize tissue culture conditions for genetic transformation of Zoysia matrella (L.) Merr., we investigated the effects of different plant growth regulators and medium supplements on plant regeneration using stolon explants excised from mature plant grown in a green house. Plant regeneration frequency was 33.3% when stolon tissues with a node were cultured on the regeneration medium supplemented with 0.5 mg/L 2,4-D and 1 mg/L of kinetin. Comparing the basal media tested, MS medium showed higher plant regeneration performance than N6 or SH medium. Addition of 5 mg/L $AgNO_3$ with 10 mg/L cysteine improved frequency of plant regeneration up to 40%. Among different carbon sources, 3% sucrose was found to show the best for regeneration frequency. This rapid and efficient plant regeneration system would be useful for using genetic transformation experiments of manilagrass without intervening callus-mediated regeneration.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.10
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• pp.1403-1411
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• 2002

The aims of this study were, firstly, to analyze the biochemical compositions of serum and follicular fluid (FF) from prepubertal gilts after PMSG (1,000 IU) treatment. The concentrations of total proteins, lipids, cholesterol, glucose and sex hormones (progesterone, $P_4$; estradiol-$17{\beta}$, $E_2$; testosterone, T) were measured. Secondary, the effects of porcine FF (pFF) addition (40% and 100%) in IVM media and different culture conditions [Exp. 1: mBMOC-2+20% porcine serum (PS), fresh IVM medium, filtered IVMconditioned medium, or rabbit oviducts; Exp. 2: mBMOC-2+20%PS or stepwise medium replacement procedures (SMRP) cocultured with or without cumulus cells] on the in vitro development (IVD) of porcine oocytes were also examined. Results showed that no significant differences were found in total protein levels between serum and pFF from different sizes (large, >7 mm; medium, ~5-7 mm; small, <3-5 mm) of follicles (75-85 and 49-90 mg/dl; p>0.05). Total lipid concentrations remained constant in serum (395-472 mg/dl), and reduced significantly in the pFF from large follicles (287 mg/dl) at 132 h after PMSG treatment when compared to those at other time points (441-480 mg/dl). Basal cholesterol levels in serum and pFF at 12 h were similar (153-161 mg/dl), but increased at 36 h (186-197 mg/dl). Basal P4 and E2 levels in serum (0.1 ng/ml and 5.5 pg/ml) were low, but increased from 0.34 ng/ml and 12.13 pg/ml at 24 h to 0.81 ng/ml and 61.70 pg/ml at 98 h, respectively, after PMSG treatment (p<0.05). P4 levels increased linearly in pFF from large follicles during 12 through 132 h (138-1,288 ng/ml). A similar increase was also observed in $E_2$ levels (22-730 pg/ml) before 60 h post PMSG treatment, and then dropped afterwards (730-121 pg/ml). The development of the oocytes fertilized in 40% pFF-medium was greater than that in 100% pFF-medium group without gonaodtropin addition (31% vs 10%, p<0.05). However, both were lower than those in mBMOC-2+20%PS and in rabbit oviducts (p<0.05). When cocultured with cumulus cell monolayers, a greater cleavage rate was observed in the group cultured in filtered IVM-conditioned medium than the SMRP group (36% vs 18%, p<0.05). A similar phenomenon was also observed in the culture without cumulus cell monolayers (33% vs 19%, p<0.05). It is concluded that neither the fresh IVM nor filtered IVM-conditioned medium has positive effect on the IVD of oocytes. Coculture with cumulus cell monolayers and the SMRP were not beneficial to the development of IVF pig oocytes.