• Restorative Dentistry and Endodontics
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• v.36 no.1
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• pp.26-36
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• 2011

Objectives: In the present study, three kinds of tissues cells (pulp, gingiva, and periodontal ligament) were investigated if those cells express MMP and TIMP when they were stimulated with neuropeptides (substance P, CGRP) or proinflammatory cytokine, TNF-$\alpha$. Materials and Methods: The cells cultured from human dental pulp (PF), gingiva (GF) and periodontal ligament were (PDLF) stimulated with Mock, SP, TNF-$\alpha$, and CGRP for 24 hrs and 48 hrs. for an RNase protection assay and Enzyme Linked Immunosorbent Assay. Cells (PF, GF and PDLF) seeded in 100 mm culture dish were stimulated with SP ($10^{-5}$, $10^{-8}\;M$) or only with medium (Mock stimulation) for 4hrs and for 24 hrs for RNase Protection Assay, and they were stimulated with CGRP ($10^{-5}\;M$) and TNF-$\alpha$(2 ng/mL) for 24 hrs and with various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for Rnase Protection Assay with a human MMP-1 probe set including MMP 1, 2, 8, 7, 8, 9, 12, and TIMP 2, 3. In addition, cells (PF, GF and PDLF) were stimulated with Mock and various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for 24 hrs and with TNF-$\alpha$(10 ng/mL) for 48 hrs, and the supernatents from the cells were collected for Enzyme Linked Immunosorbent Assay (ELISA) for MMP-1 and MMP-13. Results: The expression of MMPs in PF, GF, PDLF after stimulation with SP and CGRP were not changed compared with Mock stimulation for 4 hrs and 24 hrs. The expression of MMP-1, -12, -13 24 hrs after stimulation with TNF-$\alpha$ were upregulated, however the expression of TIMP-3 in PF, GF, PDLF after stimulation with TNF-$\alpha$ were downregulated. TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL) increased MMP-1 and MMP-12 expression in PF dose dependently for 24 hrs. Conclusions: TNF-$\alpha$ in the area of inflammation may play an important role in regulating the remodeling of dentin, cementum, and alveolar bone.

• Journal of Korean Society of Forest Science
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• v.78 no.4
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• pp.333-344
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• 1989

The objectives of this study were to investigate the growth of Gyrophora esculanta and to establish a method of tissue culture of the plant. The results obtained were as follows : 1. The Gyrophora esculanta was found growing mostly on the rock slopes of 722 m to 1915 min elevation on mountains in Korea. 2. Trees growing in the vicinity of the G. esculanta were mainly Quercus spp., Pinus thunbergii, Acer spp. and Lespedeza spp, Especially Quercus spp. was found growing in all of the study site. 3. The average Length of the rock slopes with G. esculanta growing on was 14 m and their aspects were mostly south. 4. The G. esculanta were found growing on rocks of Crystalline Schist, Quartz, Liparite, Granite, ete. Particularly they were mostly found on granites. The gradient of the rock slopes was in the range of 22-90 degrees. 5. The mean number of individuals of G. esculanta per one rock slope ranged from 14 at Mt. Bukhan to 70 at Mt. Jrri. Their mean diameter ranged from 1.8cm at Mt. Munsu to 4.6cm at Mt, Sokri. 6. The average percentage of G. esculanta with fruit body was 17.6%. The highest value was found at Mt. Cheonhwang (24.0%). 7. When the 100 segments of rhizoid of Gyrophora esculanta cultured in Detmer's medium supplemented with kinetine 5mg/l and 2, 4-D 3mg/l, n callus of microspore origins were induced from about 20% of the segments. As the induced n callus was transplanted on the six different types of rocks, it was observed that the juvenile G. esculanta grew best on granite and the development rate of G. esculanta on the granite was about 55%.

• Journal of Korean Society of Forest Science
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• v.90 no.1
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• pp.83-89
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• 2001

This study was conducted to test the Cd accumulation and Cd-tolerance of Pisotithus tinctorius(Pt). Pt was isolated from Pinus thunbergii forest in Muan, Chonnam Province in 1997. Pt was cultured on MMN medium supplemented with $CdSO_4{\cdot}5H_2O$ at the final concentration of 0, 0.2, 0.5, 2, and $10{\mu}g/m{\ell}$ for 40 days. Growth rate and tolerance index of the fungus were measured every week, while Cd concentration, superoxide dismutase(SOD), and glutathione reductase(GR) of the fungus were analyzed at the end of the culturing, Pt showed growth reduction in vitro at $2{\mu}g/m{\ell}$ Cd in the medium and almost stopped growth at $10{\mu}g/m{\ell}$ Cd. Tolerance index of Pt decreased with increasing Cd concentration. Cd concentration of Pt was the highest at $10{\mu}g/m{\ell}$ Cd. Activities of SOD did not show significant difference between Cd concentrations, but GR of Pt increased at $0.5{\mu}g/m{\ell}$ Cd, and decreased at $2{\mu}g/m{\ell}$ Cd. Consequently Pt could be called Cd accumulator with a tolerance mechanism to Cd. Their tolerance to Cd were expressed through the higher production of antioxidants such as GR. Pt may be used for revegetation and decontamination of soil polluted by heavy metals.

• Journal of Yeungnam Medical Science
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• v.12 no.1
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• pp.124-134
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• 1995

It is the most important to select optimal culture conditions to promote safe embryo growth in the technique of human in vitro fertilization and embryo transfer. It has been shown that the addition of biologic fluids, such as blood serum, of various origins, improved fertilization and early cleavage rates in numerous species. The purpose of this study is to attempt to measure developmental potential of mouse eggs fertilized and cleaved in Ham's F10 culture medium containing a chelating agent, EDTA and fetal cord serum. In this study, we selected 40 female mice and 20 male mice, and investigated optimal serum concentrations for mouse embryo growth. Two cell stage mouse embryos were cultured in Ham's F-10 medium, Ham's F-10 medium with various concentrations of EDTA, or Ham's F-10 medium with EDTA and 10% human cord serum. Developmental ratios to morula in Ham's F-10 medium containing various concentrations of EDTA and/or 10% fetal cord serum were significantly higher than in unsupplemented Ham's F-10 medium (p<0.05). Developmental ratios to blastocyst in Ham's F-10 containing 10% fetal cord serum and $50{\mu}M$ or $100{\mu}M$ EDTA were significanltly higher than in unsupplemented Ham's F-10 medium (p<0.05). Developmental ratios to morula in Ham's F-10 containing 10% fetal cord serum and $100{\mu}M$ EDTA were significanltly higher than in Ham's F-10 with 10% fetal cord serum used commonly in many human IVF centers(p<0.05). Developmental ratio to blastocyst in Ham's F-10 containing 10% fetal cord serum and $100{\mu}M$ EDTA was significanlty higher than in Ham's F-10 with $200{\mu}M$ EDTA(P<0.05). In summary, embryo development to morula and blastocyst was significanlty higher in the presence of human cord serum or EDTA than in the unsupplemented medium. The most significanly development to morula and blastocyst was obtained at Ham's F-10 medium with $100{\mu}M$ concentration of EDTA and 10% fetal cord serum. These results suggest that Ham's F-10 medium containing 10% fetal cord serum and optimal concentrations of EDTA significantly promoted early cleavage of mouse zygotes, and these will be useful as basic data for the selection of culture medium in human in vitro fertilization.

• The Korean Journal of Blood Transfusion
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• v.23 no.2
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• pp.115-126
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• 2012

Background: Umbilical cord (UC) is a promising source of mesenchymal stromal cells (MSCs). We compared the characteristics of MSCs from cryopreserved UC with those from fresh tissues, and demonstrated the possibility of UC cryopreservation for acquisition of MSCs from cryopreserved UC. Methods: Each UC was sliced into two types ($1{\sim}2mm^3$ vs. 0.5 cm), and cryopreserved in liquid nitrogen using different media (autologous cord blood plasma, aCBP vs. RPMI 1640). A fresh aliquot of $1{\sim}2mm^3$-sized UC was used as control tissue. After one week, the cryopreserved tissues were thawed and cultured. For the 0.5 cm UC, a slicing step into $1{\sim}2mm^3$ was needed. Cell count, viability, proliferative activity, and surface antigens were determined from harvested MSCs. Several growth factors (EGF, IGF-1, PDGF, TGF-${\beta}$, bFGF, and VEGF), were measured from the culture supernatant. Results: Eleven UC were enrolled in the study. Efficiencies of obtaining MSCs were higher in cryopreserved UC using RPMI 1640, compared with use of aCBP; the same result was observed for 0.5 cm sized UC, compared with $1{\sim}2mm^3$ sized UC. No difference in proliferative activity was observed between MSCs from fresh and cryopreserved UC. The amount of growth factors in culture supernatant using RPMI 1640 was larger than that of fresh tissues. Conclusion: We obtained growth factors from the supernatant as well as MSCs from cryopreserved UC. As with a cord blood bank, in the future, cryopreservation of UC for acquisition of both MSCs and growth factors would be possible in a time of need.

• FLOWER RESEARCH JOURNAL
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• v.19 no.1
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• pp.22-29
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• 2011

The experiment investigated effect of duration, temperature, and light condition during chilling treatment on growth and flowering of four Campanula species in a factorial experiment. Two parent species, Campanula punctata Lam. var. rubriflora Mak. and C. Punctata Lam., and their two $F_1$ hybrids, C. punctata Lam. ${\times}$ C. punctata Lam. var. rubriflora Mak. ('Jiknyeo') and C. punctata Lam. var. rubriflora Mak. ${\times}$ C. punctata Lam. ('Gyeonu'), were used. Plants were cultured in vitro for five weeks at $25^{\circ}C$ under about $75{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ PPFD before being chilled at 4 or $25^{\circ}C$ for 3, 6, or 9 weeks under a darkened or lighted (about $10{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ PPFD) condition. After chilling treatment, plants were transplanted to 10 cm pots filled with a commercial growing medium and were transferred to environment-controlled growth chambers and subsequently to a greenhouse to observe their reproductive growth. Growth of all species and flowering of a $F_1$ hybrid 'Jiknyeo' were affected by duration, temperature, and light condition during chilling treatment. The greatest growth and survival percentage were observed in C. punctata Lam. var. rubriflora Mak. The survival percentage was greater when plants were chilled in a lighted than darkened condition, whereas it decreased when plants were chilled more than six weeks in vitro. Among the four species tested, flowering was observed only in a $F_1$ hybrid 'Jiknyeo' with 62.5% flowering plants when it was chilled at $25^{\circ}C$ for three weeks under a lighted condition. Percent flowering plant was affected by duration, temperature, and light condition during chilling treatment. Three-week chilling at $4^{\circ}C$ under a darkened condition significantly reduced days to flowering. These results suggest that the low temperature requirement for flowering is not qualitative but quantitative in Campanula species. Further experiment with more number of plants is necessary to ascertain this conclusion.

• Journal of Food Hygiene and Safety
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• v.34 no.5
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• pp.495-501
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• 2019

Aloin [1,8-Dihydroxy-10-(${\beta}$-D-glucopyranosyl)-3-(hydroxymethyl)-9(10H)-anthracenone], is a natural anthraquinone from aloe. It has been shown to have antioxidant and anticancer effects in various types of human cancer cells, but the anticancer effects of aloin in human colorectal cancer cells HT-29 have not been elucidated. In this study, possible mechanisms by which aloin exerts its apoptotic action in cultured human colorectal cancer HT-29 cells were investigated. The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay shows that treatment with aloin (0, 100, 200, 300 and $400{\mu}M$) reduced cell viability in a concentration-dependent manner in HT-29 and showed no effects on cell proliferation in A375SM and AGS cells. In addition, it was confirmed that apoptotic body was significantly increased as shown by 4',6-diamidino-2-phenylindole (DAPI) staining, and increased apoptosis rate by flow cytometry in HT-29 cells treated with aloin (0, 200 and $400{\mu}M$). We confirmed by western blotting that aloin activated Bax (pro-apoptotic), cleaved-poly (ADP-ribose) polymerase (PARP) and caspase-3, -8 and Bcl-2 (anti-apoptotic) were not changed compared with the control. Aloin induced up-regulation of phospho-p38 and down-regulation of phospho-extracellular signal-regulated kinase (ERK)1/2. Therefore, aloin suppressed the growth inhibitory effects by the induction of apoptosis in human colorectal cancer cells and has potential as a cancer preventive medicine.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.3
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• pp.187-197
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• 2009

Objective: Phosphorylation and dephosphorylation of proteins are important in regulating cellular signaling pathways. Bead-based multiplex phosphorylation assay was conducted to detect the phosphorylation of seven proteins to maximize the information obtained from a single lysate of stage-specific mouse oocytes at a time. Methods: Cumulus-oocyte complexes (COCs) were cultured for 2 h, 8 h, and 16 h, respectively to address phosphorylation status of seven target proteins during oocyte maturation process. We analyzed the changes in phosphorylation at germinal vesicle (GV, 0 h), germinal vesicle breakdown (GVBD, 2 h), metaphase I (MI, 8 h), and metaphase II (MII, 16 h in vitro or in vivo) mouse oocytes by using Bio-Plex phosphoprotein assay system. We chose seven target proteins, namely, three mitogen-activated protein kinases (MAPKs), ERK1/2, JNK, and p38 MAPK, and other 4 well known signaling molecules, Akt, GSK-$3{\alpha}/{\beta}$, $I{\kappa}B{\alpha}$, and STAT3 to measure their phosphorylation status. Western blot analysis and kinase inhibitor treatment for ERK1/2, JNK, and Akt during in vitro maturation of oocytes were conducted for the confirmation. Results: Phosphorylation of ERK1/2, JNK, p38 MAPK and STAT3 was increased over 3 folds up to 20 folds, while phosphorylation of the other three signal molecules, Akt, GSK-$3{\alpha}/{\beta}$, and $I{\kapa}B{\alpha}$ was less than 3 folds. All of these results except for Akt were statistically significant (p<0.05). Conclusion: This is the first report on the new and valuable method measuring many phosphoproteins simultaneously in one minute sample such as oocyte lysates. All of the three MAPKs, ERK1/2, JNK, and p38 MAPK are involved in the process of mouse oocyte maturation. In addition, STAT3 might be important regulator of oocyte maturation, while Akt phosphorylation at Serine 473 may not be involved in the regulation of oocyte maturation.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.1 s.49
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• pp.97-102
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• 2005

One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.6
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• pp.852-857
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• 2010

This study was performed to explore the effect of the clay mineral illite on the improvement of bed soil and plant growth. Red pepper (Capsicum annuum L.) was used as a model vegetable crop. The experiment was performed during the whole six weeks in the glass house of the Chungbuk National University. Its seedlings were cultivated in the bed soil normally used for horticultural purpose. Of the seedlings cultured, the healthy and regular size of seed were selected and cultivated in the pots. They were treated with two forms of illite, particulate (PA) and powder (PW), at the following application rates: standard application[P1 (PA1, PW1), 1:20 (w/w)], two times[P2 (PA2, PW2), 1:10 (w/w)], and four times[P4 (PA4, PW4), 1:5 (w/w)] of standard application. Untreatment (P0) was used as a control pot. At six weeks of cultivation, their growth lengths were correspondingly increased as the application rate was increased ranging from P0, P1, P2, and to P4. Their growth length was a little greater on the application of powder illite (PW) than on the particulate illite (PA). Based on the plant analysis for the root, leaf, stem of red pepper, the uptake amounts of K, Ca, and Mg, were correspondingly increased, as the application rate was increased ranging from P0, P1, P2, and to P4, respectively. At the same application rate, their amounts taken up in the respective parts were higher on the application of PW illite than on the PA one. Especially the amounts of Ca and Mg were higher in the stem, leaf than root. Consequently, it appears that the illite treatment, especially, PW form of illite, enhance the growth of red pepper in the glass house during the whole six weeks of experiment.