• The Korea Journal of Herbology
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• v.33 no.5
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• pp.67-72
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• 2018

Objectives : Natural extracts have been extensively studied to replace single agent drugs that cause a variety of side effects. However, studies of changes to the formulation of natural extracts has not been nearly proceed. We aimed to investigate whether pharmacological stability of hyeonggaeyeongyotang gagambang (HYT) is altered by formulation changes for foaming tablet. Methods : In this study, we performed freeze - drying of HYT, which is known to have antioxidant and anti - inflammatory properties, and then changed the formulation by foaming. Results : As a result, the foaming reaction appeared normally when HYT foamed tablets were put into water, and almost all of the substances were dissolved in the aqueous solution. In addition, we confirmed using high-performance liquid chromatograph that the geniposide used as an indicator material of HYT was stable in most of the formulations. It was confirmed that the change of HYT formulation did not affect the antioxidant efficacy by the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid assay. Moreover, quantitative real-time PCR confirmed that the inhibitory effect of HYT on IL-$1{\beta}$ mRNA expression induced by lipopolysaccharides treatment in murine macrophage RAW 264.7 cells was similar in the solution of foaming tablet. Conclusions : These results suggest that the materials with various pharmacological effects can be stably maintained even when the formulation is changed by the foaming action of HYT. Our results are expected to provide important basic knowledge on formulation changes using various natural extracts.

• Molecules and Cells
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• v.45 no.8
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• pp.588-602
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• 2022

Various RNA-binding proteins (RBPs) are key components in RNA metabolism and contribute to several neurodevelopmental disorders. To date, only a few of such RBPs have been characterized for their roles in neocortex development. Here, we show that the RBP, Rbms1, is required for radial migration, polarization and differentiation of neuronal progenitors to neurons in the neocortex development. Rbms1 expression is highest in the early development in the developing cortex, with its expression gradually diminishing from embryonic day 13.5 (E13.5) to postnatal day 0 (P0). From in utero electroporation (IUE) experiments when Rbms1 levels are knocked down in neuronal progenitors, their transition from multipolar to bipolar state is delayed and this is accompanied by a delay in radial migration of these cells. Reduced Rbms1 levels in vivo also reduces differentiation as evidenced by a decrease in levels of several differentiation markers, meanwhile having no significant effects on proliferation and cell cycle rates of these cells. As an RNA binding protein, we profiled the RNA binders of Rbms1 by a cross-linked-RIP sequencing assay, followed by quantitative real-time polymerase chain reaction verification and showed that Rbms1 binds and stabilizes the mRNA for Efr3a, a signaling adapter protein. We also demonstrate that ectopic Efr3a can recover the cells from the migration defects due to loss of Rbms1, both in vivo and in vitro migration assays with cultured cells. These imply that one of the functions of Rbms1 involves the stabilization of Efr3a RNA message, required for migration and maturation of neuronal progenitors in radial migration in the developing neocortex.

• Korean Journal of Environmental Biology
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• v.29 no.3
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• pp.236-240
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• 2011

Mercury known as quicksilver, is the most common cause of heavy metal toxicity. Toxicity caused by excessive mercury exposure is now being recognized as a widespread environmental problem and is continuing to attract a great deal of public concerns. The mercury genotoxicity could be its effect on DNA repair mechanisms, which constitute the defense system designated to protect genome integrity. The objective of this study is to confirm that mercuric chloride inhibits the repair of gamma ray-induced DNA damage. The earthworm of Eisenia fetida was chosen for this study because it is an internationally accepted model species for toxicity testing with a cosmopolitan distribution. Experiments were done to identify the levels of DNA damage and the repair kinetics in the coelomocytes of E. fetida irradiated with 20 Gy gamma rays alone or with gamma rays after 40 mg $kg^{-1}$ $HgCl_2$ treatment by means of the single cell gel electrophoresis assay. The Olive tail moments were measured during 0~96 hours after irradiation. The repair time in the animals treated with the combination of $HgCl_2$ and ionizing radiation was nearly five times longer than that in the animals treated with ionizing radiation alone. Also, E. fetida exposed to mercury showed a statistically lower repair efficiency of gamma ray-induced DNA damage. The results suggest that the mercury could even have deleterious effects on the DNA repair system. Influence of mercury on the DNA repair mechanisms has been confirmed by this study.

• Journal of Radiation Protection and Research
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• v.26 no.4
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• pp.385-392
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• 2001

Environmental risk factors such as ionizing radiations, heavy metals, and pesticides can cause environmental disasters when they exist in excess. The increases in use of ionizing radiation and agricultural pesticide are somewhat related to the possibility of the disaster. The risk of radiation and pesticide was evaluated by means of the single cell gel electrophoresis (SCGE) assay on the human blood lymphocytes. The lymphocytes were irradiated with $0{\sim}2.0Gy$ of $^{60}Co$ gamma ray. Another groups of lymphocytes were exposed to various concentrations of parathion. Significantly increased tail moment, which was a marker of DNA strand breaks in SCGE assay, showed a clear dose- or concentration-response relationship. Parathion of a recommended concentration for agricultural use ($1mg {\ell}^{-1}$ ) has a strong cytotoxic effect on lymphocytes, which is equivalent to damage induced by 0.1 Gy of ${\gamma}$-ray. Furthermore, $2mg{\ell}^{-1}$ of parathion can give rise to DNA damage equivalent to that induced by 0.25 Gy at which the radiation-induced damage can start to develop into clinical symptoms. The comparative results of this study can provide an experimental basis and biological information for the prevention of environmental disaster.

• The Korea Journal of Herbology
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• v.32 no.4
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• pp.1-8
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• 2017

Objectives : Silkworm is known as immunomodulatory substances and contain various bioactive compounds such as serine, tyrosine and alanine. The aim of this study was to investigated the immunopotentiating activity of combine extract that silkworm and food materials (Eucommia ulmoides, Angelica gigas, Acanthopanax, Allium hookeri, Cinnamomum cassia, Liriope platyphylla, Curcuma longa, Achyranthes japonica, Alpinia oxyphylla, Adenophora triphylla). Methods : Among 10 kinds of food materials, to select food materials with the effect of enhancing the immune function mouse splenocyte proliferation ability was measured by 3-[4,5-dimethylthiazol-2-y]-2,5-diphenyl terazolium bromide (MTT) assay. Then, combine extract of silkworm and food materials were evaluated that mouse splenocyte proliferation ability by EZ-cytox cell viability assay. Morever, cytokines production such as IL-2, IL-4, IL10, IL12, $IFN-{\gamma}$ on mouse T lymphocyte stimulated with concanavalin A (ConA) was measured. Results : Eucommia ulmoides, Acanthopanax, Allium hookeri, Cinnamomum cassia, Liriope platyphylla has high proliferation ability of mouse splenocyte compared with Curcuma longa, Achyranthes japonica, Alpinia oxyphylla, Adenophora triphylla. The silkworm and food material combined extract has a relatively high proliferation ability of mouse splenocyte proliferation when the silkworm and food materials are used as a single material. In particularly, combined extract of silkworm and Cinnamomum cassia was stimulate cytokine production on T lymphocyte such as IL12, $IFN-{\gamma}$. Combined extract of silkworm and Liriope platyphylla was stimulate cytokine production on T lymphocyte such as IL2, IL4, IL10. Conclusion : In conclusion, the combined extract of the silkworm and Cinnamomum cassia or Liriope platyphylla may enhance immune function by regulating mouse splenocyte proliferation and stimulating cytokine production.

• Proceedings of the Plant Resources Society of Korea Conference
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• v.18 no.1
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• pp.52-60
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• 2005

The objectives of present study were to investigate the hepatoprotective and antioxidative effects of onion extracts. Primary cultures of rat hepatocytes were incubated with 1.5 mM tert-butyl hydroperoxide(t-BHP), potent oxidizing agent for liver injury for 1 hr in the presence or absence of various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring catalase, glutathione peroxidase(GSH-Px), glutathione reductase(GSH-Rd) activities as well as DNA strand breaking assay. Incubation with t-BHP alone increased GOT and LDH activities and TBARS concentration but decreased MTT reduction. Onion extracts at the concentration of 0.05 mg/ml began to decrease GOT and LDH activities induced by 1.5 mM t-BHP. Decreased MTT reduction began to be increased by onion extract at the concentration of 0.01 mg/ml. Onion extracts at the concentration of 0.01 mg/ml began to decrease TBARS concentration induced by t-BHP. Taken together, onion extracts prevented t-BHP-induced hepatocyte injury and lipid peroxidation. Catalase, GSH-Px and GSH-Rd activities of hepatocytes were significantly decreased by 1.5 mM t-BHP for 1 hr incubation. Onion extracts, on the other hand, at the concentration of 0.1 mg/ml began to prevent t-BHP-induced decrease in catalase, GSH-Px and GSH-Rd activities. Onion extracts prevented hydroxyl radical-induced single-strand breakage in dose-dependent manner when plasmid DNA was incubated with various concentrations of onion extracts in the presence of Fenton regents producing hydroxyl radical. These results demonstrate that onion extracts suppressed t-BHP-induced cytoctoxicity, decreased viability and lipid peroxidation and increased GSH-Px, GSH-Rd and catalase activities. Thus hepatoprotective and antioxidant effects of onion extract seem to be due to, at least in part, the increase in antioxidant enzyme activities as well as prevention from hydroxyl radical-induced oxidation, followed by inhibition in lipid peroxidation.

• Korean Journal of Veterinary Research
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• v.44 no.3
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• pp.379-387
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• 2004

When an organism is exposed to various toxicants chronically, reactive oxygen species(ROS) are accumulated and eventually result in several biological effects from gene expression to cell death. In the present study we investigated the oxidative damage of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin(TCDD) and/or benzo(a)pyrene (B(a)P) in C100 cells. C100 cells treated with TCDD(30 nM) and B(a)P($3{\mu}M$) underwent diverse oxidative stress as determined through thiobarbituric acid-reactive substances(TBARS) formation, DNA fragmentation, DNA single strand break(SSB) assay, immunohistochemical staining of 8-hydroxy-2'-deoxyguanosine(8-OHdG), and mRNA expressions of antioxidant enzymatic genes such as Cu/Zn-SOD gene, GPx(glutathione peroxidase 5) gene, and catalase gene. Lipid peroxidation in C100 cells was determined through measuing the formation of TBARS. For theat, the cells were pretreated with TCDD(30 nM) and/or B(a)P($3{\mu}M$) for 0.5, 1, 2 and 4 days. TBARS formation was increased in TCDD(30 nM) and B(a)P($3{\mu}M$) and mixture($30nM\;TCDD+3{\mu}M\;B(a)P$) and positive control treatment groups comparing to the controls. Mixture treatment induced more DNA fragmentation than the single treatment group at day 6. Also, SSB in all treatment groups was clearly observed when compared with the negative control group. As with the expression of antioxidant enzyme, GPx 5mRNA, B(a)P alone and mixture($30nM\;TCDD+3{\mu}M\;B(a)P$) treatment were higher comparing to those of the negative control and TCDD treatment groups. Our results suggest that exposure of C100 cells to mixture of TCDD and B(a)P leads to significant oxidative damage comparing to the exposures to the individual chemicals. Mechanisms of action are discussed. Additional studies are needed to elucidate the detailed mechanism of mixture-induced toxicity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.3
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• pp.355-362
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• 2013

This study was performed to assess the antioxidant activities and whitening effects of protopectinase enzymes and mechanical maceration from soybeans on melanin synthesis. The whitening effects of enzyme treatment and mechanical maceration were examined by an in vitro mushroom tyrosinase assay and by assessing markers in B16BL6 melanoma cells. We assessed inhibitory effects on the expression of melanogenic enzymes, including tyrosinase, tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2) in B16BL6 cells. Inhibitory effects on free radical generation were determined by measuring DPPH and hydroxyl radical scavenging activities. In DPPH radical scavenging activity, enzyme treatment and mechanical maceration had a potent anti-oxidant activity in a dose-dependent manner and significantly inhibited tyrosinase activity in vitro and in B16BL6 melanoma cells. There was also an inhibition in the expression of tyrosinase, TRP-1, and TRP-2 in B16BL6 melanoma cells. Our results show that soybean protopectinase treatment inhibits melanogenesis, with the underlying mechanism possibly due to the inhibition of tyrosinase activity and tyrosinase, TRP-1, and TRP-2 expression. We suggest that soybean protopectinase should be contained as natural active ingredients for antioxidant and whitening cosmetics.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5415-5420
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• 2013

Background: Cell cycle deregulation is a major component of carcinogenesis. The p53 tumor suppressor gene plays an important role in regulating cell cycle arrest, and mouse double minute 2 (MDM2) is a key regulator of p53 activity and degradation. Abnormal expression of p53 and MDM2 occurs in various cancers including lung cancer. Methods: We investigated the distribution of the p53 Arg72Pro (rs1042522) and MDM2 SNP309 (rs2279744) genotypes in patients and healthy control subjects to assess whether these single nucleotide polymorphisms (SNPs) are associated with an increased risk of lung adenocarcinomas in Chinese female non-smokers. Genotypes of 764 patients and 983 healthy controls were determined using the TaqMan SNP genotyping assay. Results: The p53 Pro/Pro genotype (adjusted OR = 1.55, 95% CI = 1.17-2.06) significantly correlated with an increased risk of lung adenocarcinoma, compared with the Arg/Arg genotype. An increased risk was also noted for MDM2 GG genotype (adjusted OR = 1.68, 95% CI = 1.27-2.21) compared with the TT genotype. Combined p53 Pro/Pro and MDM2 GG genotypes (adjusted OR = 2.66, 95% CI = 1.54-4.60) had a supermultiplicative interaction with respect to lung adenocarcinoma risk. We also found that cooking oil fumes, fuel smoke, and passive smoking may increase the risk of lung adenocarcinomas in Chinese female non-smokers who carry p53 or MDM2 mutant alleles. Conclusions: P53 Arg72Pro and MDM2 SNP309 polymorphisms, either alone or in combination, are associated with an increased lung adenocarcinoma risk in Chinese female non-smokers.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.6
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• pp.4328-4334
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• 2015

We determined a method to determine marine planktonic organism viability using Evan's blue, Aniline blue, and 5-choromethyfluorescein diacetate (CMFDA). The Evan's blue and Aniline blue methods produced bright blue light for dead phytoplankton and zooplankton and were the best dyes to detect dead cells. The staining efficiency of Evan's blue and Aniline blue were ${\geq}90%$ of the original field sample. However, it was difficult to test the efficiency of a ship's ballast water treatment system because detection of living cells. In contrast, the CMFDA method, which is based on measuring cell esterase activity using a fluorimetric stain, was the best dye to detect live cells of almost all phytoplankton species, and staining efficiency was 70%. The CMFDA method is similar to the fluorescein diacetate (FDA) staining method. Therefore, we estimated viability of phytoplankton species using a double-staining method by combining CMFDA and FDA to determine optimum staining efficiency. As a result, the frequency of dying cells based on the double-staining method was 95%, which was significantly higher than that of single CMDFA staining. Our results suggest that a CMDFA + FDA assay is more effective to determine survival of marine plankton and that this method was applicable to investigate the efficacy of a ship's ballast water treatment system.