Journal of the korean academy of Pediatric Dentistry
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v.26
no.2
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pp.427-436
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1999
Preservation of the remaining periodontal ligament cells on an avulsed tooth is very important to the successful outcome of replantation. HBSS is recommended as the most suitable storage medium for the avulsed tooth that cannot be replanted immediately. But their availability near the site of an accident is doubtful. The purpose of this in vitro study was to compare periodontal ligament cells stored in different storage media obtained easily on the spot. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ culture medium containing 20% FBS, at $37^{\circ}C$ 100% humidity, in a 5% $CO_2$ incubator. Cells were cultured in 96 well culture plate, $5{\times}10^4$ cells per well with ${\alpha}-MEM$ and incubated for 24 hours. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS, pasteurized milk, sterilized saline, unstimulated saliva and bench-dried state at $25^{\circ}C$ room temperature for 30, 60, 90, 120, 180 minutes respectively. And then each group was measured using MTT assay. The results were as follows. 1. Between the group of each time, there was statistically significant difference. Periodontal ligament cells viability was highest in pasteurized milk and was reduced stepwisely in sterilized saline, unstimulated saliva and bench-dried state(p<0.05). 2. between the time of each group, there was statistically significant difference(p<0.05) but was no statistically significant difference at 90-120 minutes in pasteurized milk and at 60-90 minutes and 120-180 minutes in sterilized saline(p>0.05). In conclusion, HBSS as storage medium of an avulsed tooth is not practical on the spot. Insteadily pasteurized milk can be recommended to maintain the periodontal ligament cells viability.
In most mammals, mature oocyte-cumulus complexer(OCCs) ovulate into the oviduct where fertilization by sperm takes place. However, the complex that fail to fertilize eventually undergoes degeneration while they reside in the oviduct. Yet there is no blown mechanism how both oocyte and cumulus cells degenerate. Using human follicular fluid (hFF), bovine oviductal tissue extract (BOX) and mouse OCC, the present study aimed to find how the oviduct influence the viability of the oocyte and cumulus cells in vitro. There was no difference of oocyte maturation rate between the control and BOX-treated groups. However, there was a significant difference in the survival of cumulus cells between two groups. Cumulus cells cultured in the presence of hFF alone underwent initially expansion and then they formed monolayer in the culture dish. Even after 72 hr, they proliferated well and showed fibroblast-like morphology. Cumulus cells cultured in the presence of both hFF and BOX also expanded after 24 hr, however, after 72 hr culture, they eventually detached and degenerated. Cumulus cells cultured in the BOX alone gave a similar drastic result. When the cumulus cells cultured in the presence of BOX were stained with DAPI, their nuclei showed partial condensation and fragmentation. After detailed analysis of these cells by TUNEL assay, many nuclei of them exhibited well stained spots indicating the signs of apoptosis. Based upon these observations, it is suggested that BOX might possess a factor that leads mouse cumulus cells to undergo apoptosis in vitro.
Seo, Dong-Joo;Kim, Jeong-Mi;Kim, Tae-Hyuk;Baek, Jong-Mi;Kim, Tae-Woo;Kim, Hyun-Sook;Choe, Myeon
Journal of the Korean Society of Food Science and Nutrition
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v.39
no.11
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pp.1604-1610
/
2010
We investigated the anti-obesity effects of Foeniculum fructus water extract on body weight, epididymal adipocyte size, plasma lipid levels and activities of key enzymes such as lipoprotein lipase (LPL), acyl-CoA synthetase (ACS) in high fat diet-induced obese mice. Experimental groups were normal diet group (ND), high fat diet group (HFD), high fat diet with 0.05% orlistat group (HFDO), and high fat diet with 0.5% Foeniculum fructus group (HFDF). Eleven-weeks feeding with HFD resulted in significant increase in lipid levels, body weight, liver and epididymal adipose tissue weight, compared with the ND group. Diet containing Foeniculum fructus water extract significantly reduced plasma total cholesterol, triglyceride and glucose concentrations as well as body weight, liver and epididymal adipose tissue weights. Plasma LDL cholesterol levels were significantly lower in the HFDF group than those in HFDO group. LPL activities elevated by a high fat diet were significantly decreased by Foeniculum fructus water extract administration. ACS activities decreased in the high fat diet group and markedly increased in the Foeniculum fructus water extract administered group. All things considered, Foeniculum fructus water extract efficiently inhibits the inflow of fatty acid into the cell, and activates metabolic process that uses fatty acids flowing as an energy source. Thus, Foeniculum fructus water extract may have great potential as a novel anti-obesity agent.
Purpose: Poncirus trifoliata has been reported to have anti-inflammatory, antioxidant, and immune activities. However, its anti-obesity activity and the mechanism by which the water extract of dried, immature fruit of Poncirus trifoliata (PF-W) acts are not clear. This study suggests a potential mechanism associated with the anti-obesity activity of PF-W. Methods: We measured the effect of PF-W on lipoprotein lipase (LPL) regulation using enzyme-linked immunosorbent assay (ELISA) and an activity assay. The LPL regulation mechanism was examined by reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression of biomarkers related to protein transport and by western blot for analysis of the protein expression of the transcription factor CCAAT-enhancer-binding protein ($C/EBP{\beta}$). Results: The total polyphenol and flavonoid content of PF-W was $52.15{\pm}4.02$ and $6.56{\pm}0.47mg/g$, respectively. PF-W treatment decreased LPL content in media to $58{\pm}5%$ of that in control adipocyte media, and increased LPL content to $117{\pm}3.5%$ of that in control adipocytes, but did not affect the mRNA expression of LPL. PF-W also increased the mRNA expression of sortilin-related receptor (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL, in a concentration- and time-dependent manner. Finally, cell fractionation revealed that PF-W treatment induced the expression of $C/EBP{\beta}$, a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. Conclusion: The LPL secretion and activity assay showed PF-W to be an LPL secretion inhibitor, and these results suggest the potential mechanism of PF-W involving inhibition of LPL secretion through $C/EBP{\beta}$-mediated induction of SorLA expression.
Carrot ($Daucus$$carota$ L. var. $sativa$) is one of the most widely used crops in the world. Moreover it is an important crop because of its high content of ${\beta}$-carotene, well-known as the precursor of vitamin A carotenoid. However, seed-hair which is generated in epidermal cell of seeds inhibits absorption and germination. For that reason, carrot seeds are commercialized after mechanical hair removal process. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-ATR615 OP 666-13line and hairy seed CT-ATR615 OP-CK1-9 line were constructed and expression patterns related to generation of seed-hair were analyzed by comparison of EST sequences. Differential EST sequence results between two lines were classified into FunCat functional categories based on the results of BlastX search. Higher expression quantities belonging to metabolic category were shown on short-hair seed line than hairy-seed one. Differential expression quantities between those two lines in the protein folding and stabilization, subcellular localization categories were supposed to contribute variously on the generation of seed-hair. We confirmed 50 and 59 SSR sites, and 2 SNP sites by analyzing EST sequences in two lines; thereafter, we designed SNP and SSR primer sets from these EST sequence information as a molecular marker. These markers are thought to be used in research of molecular markers for classification of carrot family and related to various traits, as well as seed-hair characteristic.
Background: It has been well documented that transient occlusion of the coronary artery causes myocardial ischemia and finally cell death when ischemia is sustained for more than 20 minutes. Extensive studies have revealed that ischemic myocardium cannot recover without reperfusion by adequate restoration of blood flow, however, reperfusion can cause long-lasting cardiac dysfunction and aggravation of structural damage. The author therefore attempted to examine the effect of postischemic reperfusion on myocardial ultrastructure and to determine the rationales for recanalization therapy to salvage ischemic myocardium. Materials and methods: Young Holstein-Friesian cows(130∼140 Kg body weight; n=40) of both sexes, maintained with nutritionally balanced diet and under constant conditions, were used. The left anterior descending coronary artery(LAD) was occluded by ligation with 4-0 silk snare for 20 minutes and recanalized by release of the ligation under continuous intravenous drip anesthesia with sodium pentobarbital(0.15 mg/Kg/min). Drill biopsies of the risk area (antero-lateral wall) were performed at just on reperfusion(5 minutes), 1-, 2-, 3-, 6-, 12-hours after recanalization, and at 1-hour assist(only with mechanical respiration and fluid replacement) after 12-hour recanalization. The materials were subdivided into subepicardial and subendocardial tissues. Tissue samples were examined with a transmission electron microscope (Philips EM 300) at the accelerating voltage of 60 KeV. Results: After a 20-minute ligation of the LAD, myocytes showed slight to moderate degree of ultrastructural changes including subsarcolemmal bleb formation, loss of nuclear matrix, clumping of chromatin and margination, mitochondrial destruction, and contracture of sarcomeres. However, microvascular structures were relatively well preserved. After 1-hour reperfusion, nuclear and mitochondrial matrices reappeared and intravascular plugging by polymorphonuclear leukocytes or platelets was observed. However, nucleoli and intramitochondrial granules reappeared within 3 hours of reperfusion and a large number of myocytes were recovered progressively within 6 hours of reperfusion. Recovery was apparent in the subepicardial myocytes and there were no distinct changes in the ultrastructure except narrowed lumen of the microvessels in the later period of reperfusion. Conclusions: It is likely that the ischemic myocardium could not be salvaged without adequate restoration of coronary flow and that the microvasculature is more resistant to reversible period of ischemia than subendocardium and subepicardium. Therefore, thrombolysis and/or angioplasty may be a rational method of therapy for coronarogenic myocardial ischemia. However, it may take a relatively longer period of time to recover from ischemic insult and reperfusion injury should be considered.
Kim, Eung-Bae;Hong, Soon-Gab;Do, Byung-Rok;Kim, Hae-Kwon;Lee, Joon-Yeong
Development and Reproduction
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v.15
no.2
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pp.99-111
/
2011
The present experiment was performed to evaluate the chondrogenic differentiation potential of human adipose tissue-derived mesenchymal stem cells (ATMSCs) in the chondrogenic induction medium (CIM) with transforming growth factor-${\beta}1$ (TGF-${\beta}1$) and to evaluate the chondrogenic differentiation of ATMSCs seeded in gelatin-chondroitinglucosamine scaffold (GCG-scaffold). ATMSCs and mouse chondrocytes were cultured in the basic medium and CIM without TGF-${\beta}1$ (CIM1) or with TGF-${\beta}1$ (CIM2) for chondrogenic differentiation potential. The chondrogenic differentiation of ATMSCs was evaluated by glycosaminoglycan (GAG) synthesis and histochemical staining. In pellet culture, GAG synthesis of ATMSCs and chondrocyte was increased in culture on 14 days, but higher in CIM1 than basic medium, especially highest in CIM2. Cartilage matrix was observed in ATMSCs cultured in CIM2 on 14 days by Safranin O and trichrome staining. In well plate culture, proliferation of ATMSCs was continuously increased in culture on 10 days and higher in CIM than basic medium. The cell adhesion rate of ATMSCs seeded in flask or scaffolds was continuously increased during culture period, but higher in scaffold than flask. GAG synthesis of ATMSCs seeded in scaffolds showed no change in control group. In the CIM groups, GAG synthesis of ATMSCs was continuously increased than control group during culture period, especially very high in CIM2 and in the GCG-scaffold was slightly higher than the gelatin scaffold (G-scaffold). The present results demonstrated that ATMSCs showed an low chondrogenic differentiation potential, compared to mouse chondrocytes for 14 days of culture. TGF-${\beta}1$ is important factor in chondrogenic differentiation of ATMSCs. Gelatin scaffold was considered to increasing the effective chondrogenic differentiation environment. ATMSCs seeded in GCG-scaffold was more effective in chondrogenesis than in G-scaffold. Conclusively, the present results demonstrated that the treatment of chondroitin and glucosamine in the scaffold was more effective to promote the cartilage matrix formation.
The Korean animation that has relatively short history compared to the Western Europe and Japan's animation started out from the non-commercial short-piece animation produced as part of advertisement animation and culture movie in the later part of 1950s. In 1960s, the culture movie animation reflecting for the Movie Act and cultural policies has hardly been mentioned in the history of Korean animation, but they are the precious cultural work produced prior to the theatrical long-piece animation. In particular, compared to the 15-second short CF animation, the short-piece animations are ranging for 4 minutes to 10 minutes as the work pieces with the historic value to measure the level of the Korean animation at that time. in 1960s, approximately 20 short-piece animation works were produced and they contained the educational contents to enlighten general public in the process of modernization policy. Those short-piece animations produced in cultural movie at the National Film Production Center of Korea had been produced not only in cell-facilitating cartoon animation, but also in paper animation and puppet animation. In this background, this thesis takes a close look to the short-piece animation works produced in the National Film Production Center of Korea in 1960s. While there was almost no studies of early short-piece animation other than CF works, it is meaningful to discover and analyze the works, and, Director Park Young-il, Director Han Sung-hak, Director Jung Do-bin, Director Shin Dong-hyun, Director Nelson Shin and others participated in the creative work process have worked as the animation directors for theater that the analysis on the works would be considered as important fundamental studies to understand the Korean animation. Under this thesis, it is intended to study the historic implication and formative characteristics around some 10 work pieces to affirm participating personnel, including directors, for the short-piece animation created by the National Film Production Center of Korea as well as the situation of time to launch the National Film Production Center of Korea in 1960s. Through this effort, it is intended to come up with the starting point to process enriched researches on non-commercial short-piece animation as well as contemplation on the Korean animation history that have been neglected in the study of the Korean animation history through such effort.
Choi, Seung Jin;Chang, Eun Deok;Kwon, Seung Oh;Kye, Dae Kon;Park, Choon Keun;Lee, Sang Won;Kang, Joon Ki
Journal of Korean Neurosurgical Society
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v.29
no.9
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pp.1215-1221
/
2000
Objective : The clinical prognosis and biological behavior of atypical and especially malignant meningiomas are well known to be worse than benign meningioma, but the degree of biological aggressiveness in each classical subtypes of benign meningioma is controversy. This study was performed to see whether there is a difference in the proliferative activity between each different histological subtypes of benign meningioma as well as atypical meningioma. Methods : Paraffin-embedded surgical specimens of 27 meningiomas, including two recurrent tumors, were studied to evaluate proliferative activity by immunohistochemical method with monoclonal antibodies to proliferating cell nuclear antigen(PCNA) and MIB-1. The specimens consisted of 8 cases of meningothelial, 9 cases of transitional, 5 cases of fibroblastic subtypes and 5 cases of atypical meningiomas. Results : Mean PCNA labeling indices of meningothelial, transitional and fibroblastic meningiomas were $4.82{\pm}5.10%$, $9.01{\pm}4.25%$ and $5.66{\pm}5.32%$, but that of atypical meningiomas was $27.62{\pm}19.67%$, noting a higher value compared to all three subtypes of benign meningiomas. Mean Ki-67 labeling indices of the above 3 subtypes were $0.43{\pm}0.85%$, $0.44{\pm}1.08%$ and $0.24{\pm}0.18%$, and that of atypical meningiomas was also revealed to be of higher value ($0.84{\pm}0.59%$). PCNA and Ki-67 labeling indices were not statistically different between histological subtypes of benign meningioma(p>0.05), but the differences of both immunolabeling between benign and atypical meningiomas were statistically significant(p<0.05). Conclusion : Immunolabeling of PCNA and Ki-67 in intracranial meningiomas reveals no prognostic difference between meningothelial, transitional and fibroblastic subtypes in classical benign meningiomas by measuring expression of PCNA and Ki-67, but it seems to be helpful in differentiating benign and atypical meningioma, later showing more proliferative activity and biological aggressiveness.
Kim, Duck-Han;Lee, Kee-Hyun;Kim, In-Ryoung;Kwak, Hyun-Ho;Park, Bong-Soo;Jeong, Sung-Hee;Ko, Myung-Yun;Ahn, Yong-Woo
Journal of Oral Medicine and Pain
/
v.35
no.3
/
pp.165-175
/
2010
Valproic acid (VPA) is a well-known anticonvulsive agent and has been used in the treatment of epilepsy for almost 30 years. VPA emerged in 1997 as an antineoplastic agent as well, when findings indicated the substance inhibited proliferation and induced differentiation of primitive neuroectocdermal tumor cells in vivo (Cinatl et al., 1997). Antitmor activity of VPA is associated with its targeting histone deacetylases. Bile acids and their synthetic derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. It has been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis-inducing activity on various cancer cells in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with the histone deacetylases inhibitor, VPA and a CDCA derivative, HS-1200 on human osteosarcoma (HOS) cells. Cell viability was evaluated by trypan-blue exclusion. Induction and augmentation of apoptosis were confirmed by Hoechst staining, flow cytometry (DNA hypoploidy and MMP change), Westen blot analysis and immunofluorescent staining. In this study, HOS cells co-treated with VPA and HS-1200 showed several lines of apoptotic manifestation such as nuclear condensations, the reduction of MMP, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF onto nuclei, and activation of caspase-7, caspase-3 and PARP whereas each single treated HOS cells did not. Although the single treatment of 1 mM VPA or $25\;{\mu}M$ HS-1200 for 48 h did not induce apoptosis, the co-treatment of them induced prominently apoptosis. Therefore our data provide the possibility that combination therapy of VPA and HS-1200 could be considered as a novel therapeutic strategy for human osteosarcoma.
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