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Mechanism underlying NO-induced apoptosis in human gingival fibroblasts

  • Hwang, In-Nam;Jeong, Yeon-Jin;Jung, Ji-Yeon;Lee, Jin-Ha;Kim, Kang-Moon;Kim, Won-Jae
    • International Journal of Oral Biology
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    • v.34 no.1
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    • pp.7-14
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    • 2009
  • Nitric oxide (NO) acts as an intracellular messenger at the physiological level but can be cytotoxic at high concentrations. The cells within periodontal tissues, such as gingival and periodontal fibroblasts, contain nitric oxide syntheses and produce high concentrations of NO when exposed to bacterial lipopolysaccharides and cytokines. However, the cellular mechanisms underlying NO-induced cytotoxicity in periodontal tissues are unclear at present. In our current study, we examined the NO-induced cytotoxic mechanisms in human gingival fibroblasts (HGF). Cell viability and the levels of reactive oxygen species (ROS) were determined using a MTT assay and a fluorescent spectrometer, respectively. The morphological changes in the cells were examined by Diff-Quick staining. Expression of the Bcl-2 family and Fas was determined by RT-PCR or western blotting. The activity of caspase-3, -8 and -9 was assessed using a spectrophotometer. Sodium nitroprusside (SNP), a NO donor, decreased the cell viability of the HGF cells in a dose- and time-dependent manner. SNP enhanced the production of ROS, which was ameliorated by NAC, a free radical scavenger. ODQ, a soluble guanylate cyclase inhibitor, did not block the SNP-induced decrease in cell viability. SNP also caused apoptotic morphological changes, including cell shrinkage, chromatin condensation, and DNA fragmentation. The expression of Bax, a member of the proapoptotic Bcl-2 family, was upregulated in the SNP-treated HGF cells, whereas the expression of Bcl-2, a member of the anti-apoptotic Bcl-2 family, was downregulated. SNP augmented the release of cytochrome c from the mitochondria into the cytosol and enhanced the activity of caspase-8, -9, and -3. SNP also upregulated Fas, a component of the death receptor assembly. These results suggest that NO induces apoptosis in human gingival fibroblast via ROS and the Bcl-2 family through both mitochondrial- and death receptor-mediated pathways. Our data also indicate that the cyclic GMP pathway is not involved in NO-induced apoptosis.

Effects of Movement Intervals on Sleep Architecture in Subjects with Periodic Limb Movements during Sleep (주기성 사지운동증의 운동간격이 수면구조에 미치는 영향)

  • Sohn, Chang-Ho;Lee, Myeong-Hi;Park, Du-Heum;Jeong, Do-Un
    • Sleep Medicine and Psychophysiology
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    • v.4 no.2
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    • pp.191-200
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    • 1997
  • Objectives : Periodic limb movements during sleep (PLMS) may cause arousals that may lead to non-restorative sleep. PLMS is characterized by long sleep latency, sleep fragmentation, frequent stage shifts, and rarity of stages 3/4 NREM sleep on polysomnography. However, controversies have existed and it still remains to be elaborated whether PLMS actually causes insomnia, since normal persons happen to have PLMS. Clinically, it would be crucial to know factors which might disturb sleep in PLMS. We became interested in Coleman's theory(1980) that invariant periodic movements disturb patients' sleep less. Though, Coleman's study seems to have been confounded by including PLMS patients with various co-morbid sleep disorders. Therefore, we attempted to study in patients only with PLMS the effects of movement patterns on sleep architecture. Methods : In 27 patients diagnosed as having PLMS only with clinical interview and nocturnal polysomnography, we studied the relationship between the movement patterns such as mean duration and variability of periodic limb movement's interval and the sleep architecture variables. Results : The shorter and the more regular the limb movement intervals were, the fewer arousals followed. The movement intervals of the older patients were shorter and more regular than the younger patients. The probability of the accompanying arousal with each limb movement increased as the duration and variance of the movement intervals increased. It decreased as the age and the frequency of limb movements increased. Among these factors the most significant one was the mean duration of the movement intervals. In other words, the shorter the movement intervals were, the less disturbed sleep was. Conclusion : PLMS frequency increases with aging but the probability of the accompanying arousal with each movement decreases with aging. Sleep-disturbing effects of PLMS depends more on the duration and variability of movement intervals than the PLMS frequency.

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Comparison of the Palatability Related with Characteristics of Beef Carcass Grade B2 and D (상등급과 등외등급 쇠고기의 기호특성 비교)

  • 문윤희;강세주;현재석;강희곤;정인철
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1152-1157
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    • 2001
  • Two kinds of samples were prepared from the loin in the carcass with grade B2 and D, which were chilled for 24 hour after slaughter. The fresh beef in this study were obtained by chilling the loin for 1 day after wrapping them. On the other hand, the chilled beef were obtained by cutting the loin by 500 g and chilling them for 30 day after vacuum packing. The experiment was carried out to compare the palatability related with characteristics of loin with grade B2 and D and to investigate the chilling effect of the loin with grade D. In the case of fresh beef, it was found that the loin with grade B2 has better meat color, but lower pH, lactic acid content, and myoglobin content, than them of the loin with grade D. Also, the loin with grade B2 has lower tenderness due to its low hardness and chewiness, and high myofibrillar fragmentation index (MFI). Furthermore, it has high monounsaturated fatty acid/saturated fatty acid (MUFA/SFA) and ATP content, and good raw meat aroma. It also shows an excellent palatability of cooked meat, although it has low cooking loss and heat shotening. On the other hand, the loin with grade D has higher chilling effect on hardness, chewiness, MFI and MUFA/SFA, than them of the loin with grade B2. However, in the case of chilled beef, the loin with grade D shows much worse tenderness, cooked meat aroma, and palatability than them of the loin with grade B2.

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Changes in the Physicochemical Properties of Spent-hen Meat during Cold and Frozen Storage (산란노계육(産卵老鷄肉)의 냉장 및 동결저장 중 물리화학적 특성 변화)

  • Gong, Yang-Sug;Moon, Yoon-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.3
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    • pp.55-61
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    • 1987
  • This study was carried out to investigate the changes in the pH, extractability of protein, ATPase activity of myofibrillar protein, myofibrillar fragmentation, freezing loss and drip loss during storage at $4^{\circ}C$ and $-20^{\circ}C$ in breast and leg muscle of spent-hen meat. pH values ill pectoral and leg muscle were lowest ell tile 1st day and 1st week during cold and frozen storage, respectively. The extractabilities of myofibrillar proteins were increased graduall during cold storage and were highest on the 1st week during frozen storage, The $Mg^{2+}-ATPase$ activities of myofibrillar proteins were highest on the 1st day and 1st week during cold and frozen storage, respectively. The myofibrillar fragmentations were greatly changed on the 1st day during cold storage and 1st week during frozen storage. Freezing losses and drip losses were increased gradually during frozen storage. pH values in breast muscle were lower than those of leg muscle, and the extractabilities, $Mg^{2+}-ATPase$ activities, fragmentations of myofibrillar proteins, and drip losses in breast muscle were higher than those of leg muscle during storage, but the patterns of the changes in both muscles were similar during storage.

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Protective Role of Tissue Transglutaminase in the Cell Death Induced by TNF-α in SH-SY5Y Neuroblastoma Cells

  • Kweon, Soo-Mi;Lee, Zee-Won;Yi, Sun-Ju;Kim, Young-Myeong;Han, Jeong-A;Paik, Sang-Gi;Ha, Kwon-Soo
    • BMB Reports
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    • v.37 no.2
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    • pp.185-191
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    • 2004
  • Tissue transglutaminase (tTGase) regulates various biological processes, including extracellular matrix organization, cellular differentiation, and apoptosis. Here we report the protective role of tTGase in the cell death that is induced by the tumor necrosis factor $\alpha$ (TNF-$\alpha$) and ceramide, a product of the TNF-$\alpha$ signaling pathway, in human neuroblastoma SH-SY5Y cells. Treatment with retinoic acid (RA) induced the differentiation of the neuroblastoma cells with the formation of extended neurites. Immunostaining and Western blot analysis showed the tTGase expression by RA treatment. TNF-$\alpha$ or $C_2$ ceramide, a cell permeable ceramide analog, induced cell death in normal cells, but cell death was largely inhibited by the RA treatment. The inhibition of tTGase by the tTGase inhibitors, monodansylcadaverine and cystamine, eliminated the protective role of RA-treatment in the cell death that is caused by TNF-$\alpha$ or $C_2$-ceramide. In addition, the co-treatment of TNF-$\alpha$ and cycloheximide ecreased the protein level of tTGase and cell viability in the RA-treated cells, supporting the role of tTGase in the protection of cell death. DNA fragmentation was also induced by the co-treatment of TNF-$\alpha$ and cycloheximide. These results suggest that tTGase expressed by RA treatment plays an important role in the protection of cell death caused by TNF-$\alpha$ and ceramide.

Large-strain Soft Sensors Using Elastomers Blended with Exfoliated/Fragmented Graphite Particles (탄성중합체와 박리 후 파쇄된 흑연입자 복합재를 이용한 대변형률 연성 센서)

  • Park, Sungmin;Nam, Gyungmok;Kim, Jonghun;Yoon, Sang-Hee
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.40 no.9
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    • pp.815-820
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    • 2016
  • An elastic polymer (e.g., PDMS) blended with EFG particles is a promising conductive composite for fabricating soft sensors that can detect an object's deformation up to or more than 50%. Here, we develop large-strain, sprayable soft sensors using a mixture of PDMS and EFG particles, which are used as a host elastomer and electrically conductive particles, respectively. A solution for a conductive composite mixture is prepared by the microwave-assisted graphite exfoliation, followed by ultrasonication-induced fragmentation of the exfoliated graphite and ultrasonic blending of PDMS and EFG. Using the prepared solutions for composite and pure PDMS, 1-, 2-, and 3-axis soft sensors are fabricated by airbrush stencil technique where composite mixture and pure PDMS are materials for sensing and insulating layers, respectively. We characterize the soft strain sensors after investigating the effect of PDMS/EFG wt% on mechanical compliance and electrical conductance of the conductive composite.

The Studies on the Development of Human Blastocyst Embryos in IVF -ET Program - II. The Development of Human Blastocyst Embryos by co-culture with Cumulus Cells (IVF-ET Program에서 Blastocyst 배아의 발생에 관한 연구 - II. 난구세포 공동배양에 의한 Blastocyst 배아의 발생)

  • Lee, Suk-Won;Yoon, San-Hyun;Yoon, Hye-Gyun;Cho, Hyon-Jin;Heo, Yong-Soo;Yoon, Hye-Jin;Park, Se-Pill;Lee, Won-Don;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.35-41
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    • 1998
  • This study was carried out to investigate the development rates of human embryos co-cultured with cumulus cells to each blastocyst stage. Human zygotes were co-cultured on cumulus cell monolayer in YS medium supplemented with 20% hFF. On day 2, if patient had four or more "good" embryos (regular blastomeres without fragmentation), embryos were further cultured for 72hrs. Blastocysts on day 5 were classified into early blastocyst (ErB), early expanding blastocyst (EEB), middle expanding Blastocyst (MEB), and expanded blastocyst (EdB) on the basis of their morphological aspects of trophectoderm cells and blastocoele. Subsequently, maximum 3 of best blastocysts were transferred in 486 cycles. The results in this study were as follows: Patients who had four or more "good" embryos on day 2 were 498 persons, but patients whose embryos could not be transferred due to failure in development to the blastocyst stage on day 5 were 12 persons (2.4%). The development rate of embryos to the blastocyst stage was 58.2% (2,885/4,957) on day 5, and the rates that developed to the ErB, EEB, MEB, and EdB stage were 15.0% (743/4,957), 14.9% (739/4,957), 14.4% (714/ 4,957), and 13.9% (689/4,957), respectively. Total 1366 blastocysts were transferred in 486 cycles (mean number=2.81). The implantation rate and the ongoing implantation rate obtained by observing the number of G-sac and FHB were 29.9% (409/1,366) and 22.5% (308/1,366), respectively. The clinical pregnancy rate was 51.2% (249/486), and the ongoing pregnancy rate' was 39.1% (190/486). Among women showing ongoing pregnancy, women with singleton were 50% (95/190), women with twin were 37.9% (72/190), and women with triplet were 12.1% (23/190). Although triplet pregnancy rate in this study was high such as 12.1%, because many blastocysts with high viability were produced in our co-culture system using cumulus cells on day 5, we really believe that a multiple pregnancy except twin should not occur by selecting good embryos for maximum two blastocyst transfer. These results demonstrate that autologous cumulus cells may be used for the production of blastocysts with high developmental competence, and the use of autologous cumulus cells to be collected easily, and to be treated conveniently at OPU must be an effective means for obtaining high implantation and pregnancy rate.

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Apoptotic Activity of Curcumin and EF-24 in HTB-41 Human Salivary Gland Epidermoid Carcinoma Cells

  • Kim, Ji-Won;Lee, Seul Ah;Go, Dae-San;Park, Byung-Sun;Kim, Su-Gwan;Yu, Sun-Kyoung;Oh, Ji-Su;Kim, Chun Sung;Kim, Jeongsun;Park, Jong-Tae;Kim, Do Kyung
    • International Journal of Oral Biology
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    • v.40 no.2
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    • pp.63-69
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    • 2015
  • Curcumin (diferuloylmethane), a constituent of turmeric powder derived from the rhizome of Curcuma longa, has been shown to inhibit the growth of various types of cancer cells by regulating cell proliferation and apoptosis. However, a need exists to design more effective analogs because of curcumin's poor intestinal absorption. EF-24 (diphenyl difluoroketone), the monoketone analog of curcumin, has shown good efficacy in anticancer screens. However, the effects of curcumin and EF-24 on salivary gland epidermoid carcinoma cells are not clearly established. The main goal of this study was to investigate the effects of curcumin and EF-24 on cell growth and induction of apoptosis in human salivary gland epidermoid carcinoma cells. Our studies showed that curcumin and EF-24 inhibited the growth of HTB-41 cells in a dose- and time-dependent manner, and the potency of EF-24 was > 34-fold that of curcumin. Treatment with curcumin or EF-24 resulted in nuclear condensation and fragmentation in HTB-41 cells, whereas the control HTB-41 cell nuclei retained their normal regular and oval shape. Curcumin and EF-24 promoted proteolytic cleavages of procaspase-3/-7/-9, resulting in an increase in the amount of cleaved caspase-3/-7/-9 in the HTB-41 cells. Caspase-3 and -7 activities were detected in viable HTB-41 cells treated with curcumin or EF-24. These results suggest that the curcumin and EF-24 inhibit cell proliferation and induce apoptosis in HTB-41 human salivary gland epidermoid carcinoma cells, and that they may have potential properties as an anti-cancer drug therapy.

Using AHP to Analyze the Evaluation Factors Related to Wildlife Passage Management (AHP기법을 활용한 야생동물이동통로의 기능개선을 위한 평가항목 분석)

  • Lee, Jin-Young;Rho, Paik-Ho;Lee, Jang-Won
    • Korean Journal of Environment and Ecology
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    • v.24 no.6
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    • pp.763-771
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    • 2010
  • The rapid increase of wildlife passage installation since the late 2000s was aimed to reduce roadkill caused by habitat fragmentation and losses related to road construction, but wildlife-vehicle collisions are now still occurred even near the wildlife passage area. This is the reason that the effectiveness of wildlife passage have not been evaluated in combination with absence of monitoring data and management strategy of the wildlife passage. The AHP method are used, in this study, to identify the evaluation factors affecting the effectiveness of the present 367 wildlife passages in a mitigation measures to reduce road effects on wildlife species. Ten evaluation factors are derived from third levels in the AHP analysis. Priority setting to identify appropriate management strategies in first level is selected among four second levels on facility, environment, wildlife species and management tool. The AHP analysis suggested that neighboring environments are the most important factor at the second level, and passage structure, harmony with natural surroundings, wildlife occurrence and monitoring of the passage are also important factors at the third levels. In summary, effective measurements of wildlife passage management is based on managing the passage with neighboring topography and natural surrounding. This is useful to establish wildlife passage management strategy in order to reduce the negative effects of roads on wildlife species.

A Study on the Mediation and Arbitration of Lease Dispute (임대차 분쟁의 조정과 중재에 관한 연구)

  • Nam, Seon-Mo
    • Journal of Arbitration Studies
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    • v.25 no.4
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    • pp.119-136
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    • 2015
  • The contracting parties must be provided a litigation scheme in order to resolve a dispute. This means taking advantage of effective measures for mediation or arbitration. A lease transaction is likely to occur mainly after a dispute. It is necessary to take the appropriate measures in advance. In general, when a variety of contracts are created, conflicts arise and disputes have to be resolved through mediation and arbitration documents, and adjustment or intervention is called for. Arbitration system is a system that is established based on the trust of the arbitral tribunal. For such system, quality education for enhancing professionalism required of the arbitrator is important. A party responding to an arbitration agreement presents a problem. The current system must ensure that there are no disadvantaged parties. However, a party must depend on an arbitration agreement that is part of the law rescue system. A litigation support by the local Bar Association must be carried out. It should be notified of the contents of the contract to select a strategy that will best resolve the conflict. In the case of lease transactions, there is a need to create a scheme to make a standard agreement that inserts an arbitration clause. Lease sale and purchase agreement or lease agreement is a form of contract that has been frequently used. Here, the arbitration agreement clause for a lawyer that will serve as arbitrator should be inserted. It is a scheme that can be activated for individuals in poor areas. In addition, it is possible to see it taking a scheme to take advantage of the lawyer system for the future of the town. The Attorney System of a town is a system that the Korean Bar Association, Legal Department has put in place since 2013. If a real estate trade dispute occurs, the role of the intermediary attorney should be to carry out his duties efficiently. In the case of real estate transaction conflicts, the lawyer of the village should be registered as the arbitrator. It is important to establish a basis of regulations through this type of real estate transaction accident analysis. Before proceeding with various adjustment systems, it is desirable to expand the arbitration region. Now we need a realtor amendment. It is the part where fragmentation of intermediary qualification is required, along with the eligibility of a subdivision.