• The Korean Journal of Physiology and Pharmacology
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• v.8 no.5
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• pp.273-280
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• 2004

Nitric oxide (NO) has been suggested to act as a mediator of cytokine-induced effects of turn over of bone. Activation of the inducible nitric oxide synthase (iNOS) by inflammation has been related with apoptotic cell death in osteoblast. YS 49, a synthetic isoquinoline alkaloid, inhibits NO production in macrophages activated with cytokines. In the present study, we investigated the molecular mechanism of YS 49 to inhibit iNOS expression in ROS 17/2.8 cells, which were activated with combined treatment of inflammatory cytokines $(TNF-{\alpha},\;IFN-{\gamma})$ and lipopolysaccharide (LPS). Results indicated that YS 49 concentration-dependently reduced iNOS mRNA and protein expression, as evidenced by Northern and Western blot analysis, respectively. The underlying mechanism by which YS 49 suppressed iNOS expression was not to affect iNOS mRNA stability but to inhibit activation and translocation of $NF-_kB$ by preventing the degradation of its inhibitory protein $I_kB_{\alpha}$. As expected, YS 49 prevented NO-induced apoptotic cell death by sodium nitroprusside. Taken together, it is concluded that YS 49 inhibits iNOS expression by interfering with degradation of phosphorylated inhibitory $_kB_{\alpha}\;(p-I_kB_{\alpha})$. These actions may be beneficial for the treatment of inflammation of the joint, such as rheumatoid arthritis.

• Korean Journal of Microbiology
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• v.47 no.3
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• pp.255-262
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• 2011

Six protein-degrading bacteria were isolated from digestive organ of Harmonia axyridis. These isolates were categorized as Staphylococcus sciuri subsp. sciuri (3 strains), Bacillus subtilis (1 strain), and Bacillus thuringiensis (2 strains) by 16S rRNA gene sequence analysis. The Staphylococcus sp. CB2-3 was selected as a protease-producing bacterium which showed the highest protease activity of 58.5 U/ml at the pH 5.0 medium. The optimal pH and temperature of protease activity were pH 5.0 and $40^{\circ}C$, respectively. This acid protease had a relatively high stability of 80% between $30-50^{\circ}C$ at broad temperature range. The opimal medium compositions of carbon, nitrogen and mineral source for cell growth and protease activity were investigated. When sorbitol (0.5%) was used as carbon source, enzyme activity was increased about 2 times than that of the basal medium. When skim milk (0.5%) was used as nitrogen source, activity was increased about 2.5 times than that of the control. Cell growth and enzyme activity were increased by mineral source such as KCl, $K_2HPO_4$, $FeSO_4$, but was completely inhibited by divalent ions such as $Co^{2+}$, $Zn^{2+}$, $Mn^{2+}$, $Cu^{2+}$.

• Korean Journal Plant Pathology
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• v.1 no.3
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• pp.199-206
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• 1985

Viroids are the smallest. well-characterized infectious agents presently known. and so far viroids have been found only in higher plants. The structures of viroid-molecules are single-stranded, covalently closed circular RNA molecules with a range of 240 to 380 nucleotides according to the various viroids. Viroids are remarkable not only as a new category of pathogen, which cause economically important diseases, but also as an excellent model system for biochemical and biophysical investigations because of their small size, relative stability and their self-replication. Four different patato spindle tuber viroid isolates, which express the different symptoms on the same host plant exchange only 2 to 6 nucleotides in the total number of 359 nucleotides, but now the mechanism of viroid pathogenicity is not explained fully. Viroid-melecules are replicated without any special viroid-associated proteins, and during the process of viroid replication oligomeric viroid-associated RNAs are detected at nuclei of viroid infected leaf tissue. The mechanism of viroid replication can now be illustrated according to a possible explanation of rolling-circle system. Although the rapid progress have been made in elucidation of the biochemical and biophysical properties of PSTV and other viroids, the mechanism of viroid replication and pathogenicity is less known and is still a matter of speculation. When these problems can be sufficiently explained, the viroid molecule could play an important role as an available vector in plant genetic engineering.

• Biomolecules & Therapeutics
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• v.20 no.6
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• pp.538-543
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• 2012

The Maillard Reaction Products (MRPs) are chemical compounds which have been known to be effective in chemoprevention. Death receptors (DR) play a central role in directing apoptosis in several cancer cells. In our previous study, we demonstrated that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal, a MRP product, inhibited human colon cancer cell growth by inducing apoptosis via nuclear factor-${\kappa}B$ (NF-${\kappa}B$) inactivation and $G_2$/M phase cell cycle arrest. In this study, (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate, a new (E)-2,4-bis(p-hydroxyphenyl)-2-butenal derivative, was synthesized to improve their solubility and stability in water and then evaluated against NCI-H460 and A549 human lung cancer cells. (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate reduced the viability in both cell lines in a time and dose-dependent manner. We also found that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate increased apoptotic cell death through the upregulation of the expression of death receptor (DR)-3 and DR6 in both lung cancer cell lines. In addition to this, the transfection of DR3 siRNA diminished the growth inhibitory and apoptosis inducing effect of (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate on lung cancer cells, however these effects of (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate was not changed by DR6 siRNA. These results indicated that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate inhibits human lung cancer cell growth via increasing apoptotic cell death by upregulation of the expression of DR3.

• Development and Reproduction
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• v.11 no.3
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• pp.205-217
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• 2007

As an initial step toward better understanding of the molecular mechanism of estrogen-dependent growth in myoma, an optimal primary cell culture condition has been developed and examined by this study. Myoma and myometrium were cultured by two different methods. Culture stability and $E_2$-responsiveness in stable culture were studied. The culture of digested tissue pieces(Method 2) was found to be a stable culture method for the myoma and myometrium showing a favorable response to estrogen. mRNA expression of PR, IGF-1 and IGF-1 receptor genes was enhanced by $E_2$. The gene responses to $E_2$ were higher in myoma compared with myometrium. Moreover, these responses were more expressive in tissues than in the surrounding cells in primary culture of normal myometrium and myoma, implying a vital role of cell communication through the extracellular matrix in maintaining the estrogen-responsiveness. The development of an improved cell culture system for myoma provides an in vitro tool to further investigate the basis of the tumor formation.

• Food Science and Preservation
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• v.23 no.6
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• pp.883-889
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• 2016

A new lactic acid bacteria with gluten-degrading activity which was isolated from salted sea foods (traditional Korea fermented food), identified as Weissella confusa (99%) by use of API kit and 16S rRNA sequencing, and designated as W. confusa. When the W. confusa cultured for 48 hours at $30^{\circ}C$ in a MRS medium containing 1% gluten, 45% of gluten was founded to be degraded. W. confusa showed 85% of survival rate at pH 3, and 94% tolerance at 0.1% oxgall, which indicates that W. confusa would survive in stomach of human. Experiments on the thermostability was confirmed that it has a stability of 70% in $50^{\circ}C$. W. confusa inhibited the growth of some pathogen, except for S. aureus. Results in this study suggest that using W. confusa for fermentation of grain flour containing gluten would be desirable to prepare the gluten-free foods needed for those who suffer from celia disease and gluten allergy.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.2 s.51
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• pp.141-146
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• 2005

Unstable cosmetic active ingredients could rapidly break down in chemical and photochemical process. Therefore, it has become a very important issue to encapsulate active ingredient for the stabilization. 7-Dehydrocholesterol (7-DHC), a precursor of vitamin $D_3$, has been shown to increase levels of protein and mRNA for heat shock protein in normal human epidermal keratinocytes. However, topical dermal application of 7-DHC is restricted due to its poor solubility and chemical unstability. In this study, 7-DHC was incorporated into nano-emulsion (NE), solid lipid nano-particle (SLN), and chitosan coated solid lipid nano-particle (CASLN), respectively. In order to prepare NE and SLN dispersion, high-pressure homogenization at temperature above the melting point of lipid was used Hydrogenated lecithin and polysorbate 60 were used as stabilizer for NE and SLN. CASLN was prepared by high speed homogenizing after adding chitosan solution to the SLN dispersion and showed positively charged particle properties. Decomposition rate of 7-DHC in NE, SLN and CASLN was studied as a function of time at different temperature. Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) studies were performed to characterize state of lipid modification. It appeared that CASLN is the most effective to stabilize 7-DHC and may be used for a useful topical dermal delivery system.

• Journal of dental hygiene science
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• v.19 no.2
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• pp.133-140
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• 2019

Background: The light-emitting diode (LED) curing light used is presumed to be safe. However, the scientific basis for this is unclear, and the safety of LED curing light is still controversial. The purpose of this study was to investigate the effect of LED curing light irradiation according to the conditions applied for the polymerization of composite resins in dental clinic on the cell viability and inflammatory response in Raw264.7 macrophages and to confirm the stability of LED curing light. Methods: Cell viability and cell morphology of Raw264.7 macrophages treated with 100 ng/ml of lipopolysaccharide (LPS) or/and LED curing light with a wavelength of 440~490 nm for 20 seconds were confirmed by methylthiazolydiphenyl-tetrazolium bromide assay and microscopic observation. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was confirmed by NO assay and $PGE_2$ enzyme-linked immunosorbent assay kit. Expression of interleukin $(IL)-1{\beta}$ and tumor necrosis factor $(TNF)-{\alpha}$ in total RNA and protein was confirmed by reverse transcription polymerase chain reaction and Western blot analysis. Results: The LED curing light did not affect the viability and morphology of normal Raw264.7 cells but affected the cell viability and induced cytotoxicity in the inflammation-induced Raw264.7 cells by LPS. The irradiation of the LED curing light did not progress to the inflammatory state in the inflammation-induced Raw264.7 macrophage. However, LED curing light irradiation in normal Raw264.7 cells induced an increase in NO and $PGE_2$ production and mRNA and protein expression of $(IL)-1{\beta}$ and $(TNF)-{\alpha}$, indicating that it is possible to induce the inflammatory state. Conclusion: The irradiation of LED curing light in RAW264.7 macrophage may induce an excessive inflammatory reaction and damage oral tissues. Therefore, it is necessary to limit the long-term irradiation which is inappropriate when applying LED curing light in a dental clinic.

• Journal of Life Science
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• v.34 no.8
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• pp.576-587
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• 2024

In this study, diverse bacterial strains were isolated from fermented foods to screen those with antibacterial activity. Among them, one strain, identified as Bacillus vallismortis MCBL 1012 through 16S rRNA gene sequence analysis, was selected for its bacteriocin production. The culture supernatant of B. vallismortis MCBL 1012 showed antibacterial activity, mainly against Gram-positive bacteria. Scanning electron microscopy (SEM) revealed that bacteriocin treatment led to cellular content leakages in Listeria monocytogenes KCCM 40307, Enterococcus faecium KCCM 12118, and Streptococcus mutans KCTC 3065. PCR analysis confirmed B. vallismortis MCBL 1012 harbored subtilosin A gene (sbo A). Antibacterial activity was decreased by proteolytic enzymes like proteinase K, subtilisin A, and α-chymotrypsin. The bacteriocin demonstrated stability at 40℃ and 60℃ for 120 min, and up to 80℃ for 60 min, with rapid activity loss at 100℃. It retained full antibacterial activity within a pH range of 4.0 to 8.0 and was not affected by up to 100% organic solvents like ethanol, methanol, acetonitrile, and tetrahydrofuran. Nevertheless, activity decreased with more than 40% isopropanol and 80% acetone. Most tested inorganic salts and detergents had no effect on antibacterial activity except, CuSO₄ and NiSO₄ at specified concentrations. The bacteriocin exerted its antibacterial effect through bactericidal action against L. monocytogenes KCCM 40307. The bacteriocin was purified by ammonium sulfate precipitation, DEAE anion exchange chromatography, and RP-HPLC. The purification resulted in a final yield of 0.03% and a 283.7-fold increase in specific activity. MALDI-TOF MS analysis determined the exact molecular weight of purified bacteriocin to be 3,326.1 Da.

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.960-968
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• 2003

A bacterial isolate showing a strong endochitosanase activity was isolated from soil and then characterized. The isolate was identified and designated as Bacillus cereus P16, based on morphological and biochemical properties, assimilation tests, cellular fatty acids pattern, along with 16S rRNA gene sequence. The optimized medium for producing extracellular chitosanase in a batch culture contained 1% tryptone, 0.5% chitosan, and 1% NaCl (pH 7.0). Powder chitosan and tryptone served the best as carbon and nitrogen sources, respectively, for the chitosanase production. Chitosanase activity was the highest when culture was completed at $37^{\circ}C$ among various temperatures ($20-42^{\circ}C$) tested in a shaking incubator (200 rpm). The levels of chitosanase activity in the culture fluid were 2.0 U/ml and 3.8 U/ml, respectively, when incubated in a flask for 60 h and in a jar fermenter for 24 h. The culture supernatant showed a strong liquefying activity on the soluble chitosan. The viscosity of 1% chitosan solution, that was incubated with the culture supernatant, was rapidly decreased, suggesting the secretion of endochitosanolytic enzymes by P16. The culture fluid revealed six endo-type chitosanase isozymes, two major (38 and 45 kD), and four minor (54, 65, 82, and 96 kD) forms by staining profile. The crude enzymes were very stable, and full activity was maintained for 4 weeks at $4^{\circ}C\;or\;-20^{\circ}C$ in the culture supernatant, suggesting a highly desirable stability rate for making an industrial application of the crude enzymes. The supernatant also cleaved the insoluble chitosan powder, but the hydrolysis rate was much lower. The enzymic degradation products of chitosan contained $(GlcN)_n$ (n=2-8). The concentration of chitosan in the reaction mixture of the crude enzyme affected the chitooligosaccharides composition of the hydrolysis products. When the higher concentration of chitosan was used, the higher degree of polymerized chitooligosaccharides were produced. By comparison with other commercial chitosanase preparations, P16 was indeed found to be a valuable enzyme source for industrial production of chitooligosaccharides from chitosan.