• Journal of Oral Medicine and Pain
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• v.34 no.3
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• pp.261-276
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• 2009

Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

• Tuberculosis and Respiratory Diseases
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• v.63 no.6
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• pp.497-506
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• 2007

Background: The present investigation was performed in rats and isolated human neutrophils in order to confirm the presumptive role of the positive feedback loop of cytosolic phospholipase $A_2$ ($cPLA_2$) activation by plateletactivating factor (PAF). Methods: The possible formation of the positive feedback loop of the $cPLA_2$ activation and neutrophilic respiratory burst was investigated in vivo and in vitro by measurement of the parameters denoting acute lung injury. In addition, morphological examinations and electron microscopic cytochemistry were performed for the detection of free radicals in the lung. Results: Five hours after intratracheal instillation of PAF ($5{\mu}g/rat$), the lung leak index, lung myeloperoxidase (MPO) activity, the number of neutrophils and the concentration of cytokine-induced neutrophil chemoattractant (CINC) in bronchoalveolar lavage fluid were increased by PAF as compared with those of control rats. The NBT assay and cytochrome-c reduction assay revealed an increased neutrophilic respiratory burst in isolated human neutrophils following exposure to PAF. Lung and neutrophilic $cPLA_2$ activity were increased following PAF exposure and exposure to hydrogen peroxide increased $cPLA_2$ activity in the lung. Histologically, inflammatory findings of the lung were observed after PAF treatment. Remarkably, as determined by $CeCl_3$ cytochemical electron microscopy, increased production of hydrogen peroxide was identified in the lung after PAF treatment. Conclusion: PAF mediates acute oxidative lung injury by the activation of $cPLA_2$, which may provoke the generation of free radicals in neutrophils.

• Tuberculosis and Respiratory Diseases
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• v.45 no.6
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• pp.1214-1222
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• 1998

Background : The intrapleural hypofibrinolysis is caused by mainly excessive concentration of pleural plasminogen activator inhibitor-1 antigen(PAI-1 Ag), which binds tissue type plasminogen activator. In pleural inflammation induced by sclerosing agents for pleurodesis, levels of pleural PAI-1 antigen increase in relation to decreasing D-dimer levels. It has been known that the pleural mesothelial cells have the capability of secreting PAI-1 Ag in response to inflammation in vivo. Therefore, we estimated whether pleural inflammation changes the balance between fibrinolytic and coagulative properties in exudative pleural effusions. Method : The thirty cases was included in our study. We determined the pleural levels of glucose, lactic dehydrogenase(LDH), pH and the counts of white blood cell(WBC), polymorpho leukocyte(PMN), lymphocyte as the parameters of pleural inflammation and cellular components of pleural fluid. The plasma level of fibrinogen in fluid and the neutrophil count in blood were determined. The levels of D-dimer, PAI-1 Ag and thrombinantithrombin III complex(TAT) were determined by ELISA(Behring, Marburg, Germany). Result : The causes of pleural effusion were as following : tuberculous in 14 cases, malignant in 10 cases and parapneumonic in 6 cases. The levels of pleural D-dimer, PAI-1 Ag and TAT was significantly higher than that of plasma(p<0..001). The severity of pleural inflammation did not correlated with pleural D-dimer, PAI-1 Ag, TAT and their plasma levels. But the level of pleural TAT correlated with pleural WBC and lymphocyte count. Conclusion : We found that the severity of pleural inflammations did not correlated with pleural D-dimer, PAI-1 Ag, TAT and the possibility of local production of PAI-1 antigen is present.

• Korean Journal of Medicinal Crop Science
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• v.13 no.6
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• pp.217-225
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• 2005

This study was carried out to investigate the genotoxicity in comet and in vitro micronucleus assay and mutagenicity in Ames test of the extracts from leaves and stem of Morus alba L. The samples showed a very weak cytotoxicity on the NIH/3T3 cells by SRB assay. The cell viability of the extracts and fractions from leaves and stems of Morus alba L. was 80% over at $500\;{\mu}g/ml$, and that of the chloroform fractions from leaves and stems showed lower than others. The genotoxicity at $250\;{\mu}g/ml$ of 100% EtOH and water extracts on the NIH/3T3 cells in comet assay was about 40% compared to positive control, and most fractions from 100% EtOH extract of the leaves showed stronger genotoxicity than that offractions from the stem. The genotoxicity with S-9 mix in vitro micronucleus assay of the 100% EtOH and water extracts form Morus alba L. did not indicate any significant difference as compared with control group. The cytokinesis-binucleated cells were showed in the hexan, chloroform, ethylacetate and butanol fractions from the extract of the leaves without S-9, and sample with S-9 showed CB cells in the chloroform fraction from the leaves. In the Ames test, the water and 100% ethanol extracts of Morus alba L. did not have a strong mutagenicity in TA98 and TA100, but the fractions of organic solvents of the ethanol extract had $10{\sim}26%$ of mutagenicity on the TA100 strain.

• Journal of Life Science
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• v.29 no.4
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• pp.484-491
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• 2019

Proanthocyanidins are naturally occurring polyphenolic compounds abundant in many vegetables, plant skins (rind/bark), seeds, flowers, fruits, and nuts. Numerous in vitro and in vivo studies have demonstrated myriad effects potentially beneficial to human health, such as antioxidation, immunomodulation, DNA repair, and antitumor activity. Among immune cells, macrophages are crucial players in a variety of inflammatory responses to environmental conditions. However, it has been widely reported that macrophages cause chronic inflammation and are involved in a variety of diseases, such as obesity, diabetes, metabolic syndrome, and cancer. In this study, we report the suppressive effect of proanthocyanidins via the heme oxygenase-1 (HO-1)-related system, on the immune response of the LPS-stimulated mouse macrophage cell line RAW264.7. Increased HO-1 expression at mRNA and protein levels were found in proanthocyanidins-treated RAW264.7 cells. Further, proanthocyanidins enhanced nuclear factor-erythroid 2-related factor 2 translocation into the nucleus. RAW264.7 cells were treated with lipopolysaccharide (LPS) with or without proanthocyanidins, and inflammatory mediator expression levels were assessed. Proanthocyanidins treatment resulted in the attenuation of nitric oxide production and inducible nitric oxide synthase expression in LPS-stimulated RAW264.7 cells. In addition, mRNA and protein expression of proinflammatory cytokines, such as tumor necrosis factor-${\alpha}$ and interleukin-6, was inhibited by proanthocyanidins treatment in LPS-stimulated RAW264.7 cells. These findings support proanthocyanidins as a promising anti-inflammatory agent.

• Journal of Life Science
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• v.31 no.1
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• pp.37-46
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• 2021

The effects of freeze-dried powder from fresh and black garlic hot water extracts on the lipid metabolism in Sprague-Dawley rats fed a high-cholesterol diet were analyzed. The experimental group was classified into the normal group (NG), the high-fat (HF) and high-cholesterol diet group (CG), the HFC and 1% fresh garlic hot water extract powder-added diet group (FGEG), and the HFC and 1% black garlic hot water extract powder-added diet group (BGEG), respectively. The serum total lipid content was 381.52±7.30 mg/ml and 368.80±4.40 mg/ml in the FGEG and the BGEG, respectively, and was significantly lower than that of the CG. The total cholesterol and triglyceride contents of the FGEG and BGEG were also significantly lower than that of the CG. The high-density lipoprotein (HDL) cholesterol was significantly higher, and the low-density lipoprotein (LDL) and the very low-density lipoprotein (VLDL) cholesterol content was lower in the FGEG and BGEG than in the CG. The serum ALT and AST activities were significantly lower than those of the CG, and especially the BGEG was lower. The total cholesterol content and the triglyceride levels of the liver tissue were 36.0% and 14.3% lower in the BGEG than in the CG, respectively. The thiobarbituric acid reactive substance (TBARS) concentrations in the serum and the liver tissue were higher in the CG than in the FGEG and BGEG, but there was no difference between them. Based on these results, garlic extract powders significantly reduced the lipid profile and increased the antioxidant activity in rats in vivo. The black garlic hot water extract powder was more effective than raw garlic because of the total number of phenolic compounds and browning substances in the black garlic.

• Journal of Life Science
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• v.31 no.11
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• pp.1028-1036
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• 2021

Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.

• Journal of agriculture & life science
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• v.50 no.2
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• pp.125-138
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• 2016

To assess the industrial possibility of mixed-extracts containing Hovenia dulcis Thunberg and 12 different botanical ingredients, a protective effect was confirmed in the chronic ethanol-induced the liver, brain, and blood injury in mouse. Blood glucose levels of the normal control group(NG) and ethanol administration group(EG) were respectively 119.43mg/dL and 305.25mg/dL, and the mixed-extracts administration group(100, 200mg/kg body weight + 25% ethanol 5g/kg body weight respectively; ME100 & ME200) were decreased to 272.76mg/dL and 234.60mg/dL. Blood ethanol contents were decreased in ME100 and ME200(3.85mg/dL, 3.08mg/dL) compared to EG(4.08mg/dL), and blood acetaldehyde contents were also decreased in ME(15.76mg/dL, 15.16mg/dL) compared to EG(18.72mg/dL). The contents of hepatotoxic indicators such as glutamine pyruvic transaminase(GPT) and glutamic oxaloacetic transaminase (GOT), nephrotoxic indicators such as blood urea nitrogen(BUN), and creatine(CRE), and total cholestero(TCHO), and triglyceride(TG) in mouse blood serum were significantly decreased in the ME compared to EG. The acetylcholinesterase(AChE) activity of ME(109.00% and 108.47%, respectively) in mouse brain tissues was decreased in ME compared to EG(116.10%). Finally, ME was remarkable in vivo antioxidant activities in the mouse liver and brain tissues by superoxide dismutase(SOD), oxidized glutathione(GSH)/total GSH ratio and the malondialdehyde (MDA) assay. Therefore, the mixed-extracts was considered to be effective a high value food with protective effect against chronic ethanol traetment-induced cytotoxicity in liver and brain tissues.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.5
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• pp.669-676
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• 2010

This study was performed to investigate the effect of Brassica rapa (BR) sprouts on weight reduction and cholesterol-lowering action in rats fed high fat diet for 4 weeks. Weight-matched male Sprague-Dawley rats were assigned to four groups according to dietary fat levels (10% or 20% of diet wt.). Experimental groups were normal diet group (N), high fat diet group (HF), high fat diet with 5% BR sprouts powder group (HF-BRL), and high fat diet with 10% BR sprouts powder group (HF-BRH). The body weight gain was increased in HF group, but gradually decreased to the corresponding level of the N group fed BR sprouts powder. The concentrations of serum LDL-cholesterol, atherogenic index and cardiac risk factor tended to decrease in the BR sprouts powder fed groups compared with the HF group. However, HDL-cholesterol concentration in serum decreased in the HF group and markedly increased in the BR sprouts powder fed groups. Concentrations of triglyceride and total cholesterol in liver were also markedly decreased in the BR sprouts powder fed groups. Triglyceride concentrations of epididymal and mesenteric adipose tissues in the BR sprouts powder fed groups were also decreased compared with the HF group. These results indicate that BR sprouts powder may reduce fat accumulation and body weight, and have cholesterol-lowering effect.

• Journal of The Korean Society of Grassland and Forage Science
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• v.5 no.2
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• pp.127-135
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• 1985

The effects of morphological development and environmental temperature on synthesis and accumulation behavior of cell-wall constituents were studied in maize cv. Blizzard and sorghum cv. Sioux and Pioneer 931 at Muenchen Technical University from 1979 to 1981. Various growth stages of maize and sorghum plants were grown on field and phytotron at 4 temperature regimes of 30/25, 25/20, 28/18 and 18/8 degree C and mid-summer sunlight over 13-hour days. The results are summarized as follow: 1. Cell-wall constituents in sorghum and maize plants were shown to have a great synthesis rates at early growth stage from growing point differentiation to final leaf visible. The highest concentration of cell wall contents were found at heading stage with 52-54% and 64-68% of neutral detergence fiber, and 30% and 45% of acid detergence fiber foe maize and sorghum, respectively. 2. The structural carbohydrates, cellulose and hemicellulose, were found as a main components of cell-wall constituents. Cellulose were mainly accumulated in stalks, while hemicellulose were an important cell wall components in leaves and panicle. 3. Synthesis rates of cell-wall constituents and non-strnctural carbohydrates were associated with increasing of temperature. Reserved carbohydrates such as fructosan, mono - and dissaccharose in plant were, however, declined when the temperature exceeded 30 deg C, during the accumulation of cellulose, hemicellulose and lignin were increased continuously. 4. Cell-wall constituents lowered digestibility and net energy accumulation in sorghum and maize plants. In a in vitro and in vivo trial, it was found a negative correlation between digestion dry matter and cell wall constituents, especially cellulose and lignin.