• Journal of Oral Medicine and Pain
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• v.26 no.4
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• pp.319-329
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• 2001

Endothelin-1 (ET-1) is a recently discovered potent vasoconstrictive peptide. It was first identified in vascular endothelial cells. ET-1 is a 21-amino acid peptide and elicits systemic effects such as stimulation of the production of atrial natriuretic peptide and release of aldosterone and corticosterone. In this study, to examine the role of ET-1 in the bone metabolism, effect of ET-1 on the proliferation and activity of osteoblastic cells was studied using HOS cells as osteoblast model. ET-1 dose-dependently increased the cell proliferation as determined by cell counting and MTT reduction assay after 48hr treatment. Alkaline phosphatase activity was inhibited by ET-1 and showed significant inhibition by 50 and 100 nM ET-1. ET-1 increased NBT reduction by HOS cells dose-dependently showing that ET-1 may increase the superoxide production by osteoblasts. Nitrite concentration in the media of HOS cell culture without cytokine stimulation was negligible and unaffected by ET-1 after 48hr treatment. Finally, after collection and concentration of conditioned media, gelatinase activity produced by HOS cells was determined by zymography. HOS cells can produce and secrete the gelatinase (gelatinase A type as determined by molecular weight of about 65,000) into culture media, however, ET-1 had no effect on the gelatinase activity. These findings suggest that ET-1 may have diverse effects on the proliferation and differentiation of osteoblasts, therefore, it may play an important role in bone metabolism.

• BMB Reports
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• v.39 no.6
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• pp.722-729
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• 2006

Recently, we have reported that 3-hydrogenkwadaphnin (3-HK), a diterpene ester isolated from Dendrostellera lessertii (Thymealeaceae), is very effective against leukemia cell lines without any detectable effects on normal cells (Moosavi et al., 2005b). In this study, we report that 3-HK induces $G_1$ cell-cycle arrest, differentiation and apoptosis in APL NB4 cell line. Indeed, the drug between 24 to 96 h induced 7-65% growth inhibition of NB4 cells. Cell viability was also decreased by 2-55% between 24 to 96 h treatments with the drug, respectively. These effects of the drug were also dose-dependent. According to flow cytomtry results, 3-HK (15 nM) induced a significant G1-arrest up to 24 h which was consequently followed with appearance of sub-$G_1$ peak at 72 to 96 h. Hoechst 33258 staining and DNA fragmentation assays confirmed the occurrence of apoptosis among the treated cells. On the other hand, NBT reducing assay, Wright-Giemsa staining, phagocytic activity and expression of cell surface markers (CD11b and CD14) confirmed that the inhibition of proliferation is associated with differentiation especially toward macrophage-like morphology. Interestingly, 3-HK at 5 and 10 nM enhanced the effects of all-trans retinoic acid (ATRA) in NB4 cells. Based on these results, 3-HK might become an ideal candidate for treatment of APL patients pending full exploration of its biological functions.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.644-653
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• 2024

Considering the emergence of various infectious diseases, including the coronavirus disease 2019 (COVID-19), people's attention has shifted towards immune health. Consequently, immune-enhancing functional foods have been increasingly consumed. Hence, developing new immune-enhancing functional food products is needed. Pinus densiflora pollen can be collected from the male red pine tree, which is commonly found in Korea. P. densiflora pollen extract (PDE), obtained by water extraction, contained polyphenols (216.29 ± 0.22 mg GAE/100 g) and flavonoids (35.14 ± 0.04 mg CE/100 g). PDE significantly increased the production of nitric oxide (NO) and reactive oxygen species (ROS) but, did not exhibit cytotoxicity in RAW 264.7 cells. Western blot results indicated that PDE induced the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. PDE also significantly increased the mRNA and protein levels of cytokines and the phosphorylation of IKKα/β and p65, as well as the activation and degradation of IκBα. Additionally, western blot analysis of cytosolic and nuclear fractions and immunofluorescence assay confirmed that the translocation of p65 to the nucleus after PDE treatment. These results confirmed that PDE increases the production of cytokines, NO, and ROS by activating NF-κB. Therefore, PDE is a promising nutraceutical candidate for immune-enhancing functional foods.

• 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.

• Food Science and Preservation
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• v.19 no.3
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• pp.455-461
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• 2012

Obesity, a strong risk factor for the development of chronic diseases, is characterized by an increase in the number and size of adipocytes differentiated from precursor cells, preadipocytes. Recent research suggests that increased reactive oxygen species (ROS) production in 3T3-L1 adipocyte facilitates adipocyte differentiation and fat accumulation. This study was to investigate whether reduced ROS production by Sargassum micracanthum extract (SME) could protect the development of obesity through inhibition of adipogenesis. 3T3-L1 preadipocytes were treated SME for up to 8 days following standard induction of differentiation. The extent of differentiation reflected by amount of lipid accumulation and ROS production was determined by Oil red O staining and nitroblue tetrazolium (NBT) assay. Treatment of SME significantly inhibited ROS production and adipocyte differentiation that is depend on down regulation of NADPH oxidase 4 (NOX4), a major ROS generator, and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$), a key adipogenic transcription factor. These results indicate that SME can inhibit adipogenesis through a reduced ROS level that involves down-regulation of NOX4 expression or via modulation of adipogenic transcription factor.

• YAKHAK HOEJI
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• v.54 no.4
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• pp.215-225
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• 2010

To investigate effects of pine essential oil isolated by steam distillation from Pinus densiflora needles (PN) and twigs (PT) on anti-oxidant and anti-aging activities, the experiments were carried out to determine anti-oxidant and antiaging activities on DPPH radical scavenging activity, NBT/xanthine oxidase-superoxide scavenging activity, silica-induced intracellular $H_2O_2$ and hydroxyl radical generation in RAW264.7 cells, hyaluronidase and elastase activities in vitro. Essential oil of PN and PT were contained 0.225 and 0.176 (w/v) %, respectively. PN was contained with 30 kinds of essential oil and its major constituent is $\alpha$-pinene (21.5%). Further PT was contained with 40 kinds of essential oil and its major constituent is $\beta$-pinene (22.4%) in GC/MS assay. Other essential oils of PN were $\beta$-pinene, camphene, myrcene, limonene, terpinolene, $\alpha$-terpineol, bornyl acetate, $\alpha$-copaene, caryophellene and humulene and PT were $\alpha$-pinene, camphene, phellandrene, limonene, terpinolene, $\alpha$-terpineol, bornyl acetate, $\alpha$-copaene, caryophellene and humulene. The essential oil of PT have more active than that of PN in anti-oxidant activity which has significant DPPH radical and superoxide scavenging activity, and significant inhibitory activities on silica-induced intracellular $H_2O_2$ and hydroxyl radical generation, as well as, significantly inhibited elastase and hyaluronidase activities. Further, phellandrene, myrcene and $\alpha$-pinene have DPPH radical and superoxide scavenging activities, $\beta$-pinene, terpinolene, myrcene and phellandrene inhibited silica-induced intracellular $H_2O_2$ and hydroxyl radical generations. And also phellandrene and $\beta$-pinene inhibited hyaluronidase and elastase activities. In conclusion, the essential oils isolated from PN and PT have anti-oxidant and anti-aging activities.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.349-357
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• 2014

In this study, the shoot clumps extract of tissue-cultured Raoulia australis using the bioreactor culture system was tested for use a natural cosmetic ingredient. Tissue-cultured R. australis shoot clumps extract was tested anti-oxidant and anti-inflammatory activity for a cosmetic application. R. australis is a wild herbaceous plant of the asteraceae growing in New Zealand and Australia. Previous studies have reported anti-viral activity of the inhibitory effects for the growth of viruses induced meningitis, bronchitis and respiratory diseases but other biological effects are unknown. The shoot clumps extract of tissue-cultured R. australis showed higher anti-oxidant effect and anti-inflammatory effect than the natural R. australis extract. In DPPH, NBT and ABTS assay, the shoot clumps extract of tissue-cultured R. australis enhanced radical scavenging activity (up to 10~25% at $50{\mu}L/mL$) more than the natural R. australis extract. Also, the shoot clumps extract of tissue-cultured R. australis inhibited expression of iNOS and COX-2 protein in LPS-stimulated Raw 264.7 macrophages more than the natural R. australis extract. From this study, the shoot clumps extract of tissue-cultured R. australis displayed strong possibility as a new natural cosmetic ingredient for skin-care products.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.6
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• pp.460-465
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• 2006

An 8-week feeding trial was conducted to investigate the effects of dietary supplementation with probiotics as a feed additive for Juvenile olive flounder (Paralichthys olivaceus). Three experimental diets supplemented with Bacillus polyfermenticus (BP), Bacillus licheniformis (BL), or Bacillus polyfermenticus plus Saccharomyces cerevisiae, (BP+SC) at $1.0{\times}10^7CFU/kg$ diet on a dry-matter basis were prepared. The basal diet was used as a control. After the 8-week feeding trial, the respiratory burst activity (NBT assay) of fish fed the BP + SC diet was significantly higher than that of fish fed the control diet. Fish fed the BP, BL and BP + SC diets had significantly lower cumulative mortality than did fish fed the control diet after the third day of the challenge test (P<0.05). However, there were no significant differences among fish fed the experimental diets in weight gain, feed efficiency, protein efficiency ratio, hematosomatic index, condition factor, survival rate, or Iysozyme activity. Results could suggest that dietary B. polyfermenticus, B. licheniformis, and B. polyfermenticus +S. cerevisiae enhance nonspecific immunity and disease resistance in juvenile olive flounder.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.172-178
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• 2013

The strains were added to the feed in the concentration of $10^3$, $10^5$, and $10^7$ CFU/kg and 2% of fishes were given the feed twice a day (8 AM and 5 PM) for 12 weeks. In result of the nonspecific immune response study to examine Respiratory burst activity, Lysozyme activity and Phagocytosis activity every two weeks until the end of the study, all test samples showed greater activities than control samples and improved immune activity with Bacillus sp. IS-2. The mortality test performed by artificial infection using Streptococcus iniae, a pathogenic bacterium, after the completion of this study also showed over 55% greater survival rate in all test samples. In result of performing PCR using the universal primer to verify that the probiotic stays in the intestines of the fishes, all test samples showed PCR product of 1,465 bp. Based on the above findings, it was concluded that Bacillus sp. IS-2 in the feed improved farmed flatfish's immune system and resistance against diseases as the probiotics. Also, the physiological indicators discovered by this study would be useful for identifying the mechanisms of probiotics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.4
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• pp.259-268
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• 2008

In this study, the antioxidative effects, inhibitory effects on tyrosinase and elastase and components of Castanea crenata leaf were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl radical, DPPH) scavenging activity ($FSC_{50}$) of extract / fractions of Castanea crenata left was in the order: 50% ethanol extract ($13.6{\mu}g/mL$) < ethyl acetate fraction (6.2) < aglycone fraction (2.1). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$ of extract / fractions from Castanea crenata leaf extract / fractions on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was in the order: aglycone fraction (0.8) < 50% ethanol extract (0.5) < ethyl acetate fraction (0.3). The scavenging activity ($IC_{50}$ for ${O_2}^{{\cdot}\;-}$ (superoxide anion radical) generated by NBT method was in the order: ethyl acetate fraction (145.5) < aglycone fraction (65.5). The protective effects on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction exhibited the most prominent cellular protective effect (${\tau}_{50}$, $191.9{\pm}12.2\;min$ at $10{\mu}g/mL$). The inhibitory effect of aglycone fraction ($9.1{\mu}g/mL$) on elastase was higher than oleanolic and ($13.7{\mu}g/mL$). And the inhibitory effect of aglycone fraction ($21.6{\mu}g/mL$) on tyrosinase was higher than arbutin ($226.2{\mu}g/mL$). But 50% ethanol extract rarely exhibited the inhibitory activity on tryosinase and elastase. Flavonoids were contained in Castanea crenata left (96.3 mg / 100 g dried Castanea crenata leaf). And flavonoids contained in ethyl acetate fraction were kaempferol, quercetin, quercitrin, and so on. Quercitrin is the most abundant component. These results indicate that extract / fractions of Castanea crenata can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging free radical and ROS, Castanea crenata leaf extract/ fractions could be used as new cosmeceutical for whitening and anti-wrinkle products.