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Immunomodulatory activities of crude polysaccharide fraction separated from Perilla frutescens Britton var. acuta Kudo (자소엽(Perilla frutescens Britton var. acuta Kudo) 조다당류 추출물의 면역활성 효과)

  • Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.49 no.5
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    • pp.559-566
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    • 2017
  • This aim of this study was to examine the immunomodulatory activities of crude polysaccharides from Perilla frutescens Britton var. acuta Kudo (PCP) in mouse bone marrow-derived dendritic cells (BMDC) and splenocytes. The immunomodulatory activity was determined by cell viability, nitric oxide (NO) production, cell surface marker expression (CD 80/86 and MHC class I/II), and cytokine production in BMDC, and cell viability, and cytokine production in splenocytes. Cell proliferation and cytokine production (tumor necrosis factor; TNF-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, and IL-12) tested in BMDC were significantly increased by PCP treatment. Additionally, the cell surface markers (CD 80/86, MHC class I/II) were highly increased by PCP treatment. For cytokine production in splenocytes, PCP treatment significantly increased the production of Th 1 cytokines [IL-2 and interferon (IFN)-${\gamma}$], but not Th 2 cytokines (IL-4). Therefore, PCP can induce immune cell activation and is a potential candidate for the development of nutraceuticals to boost the immune system.

Preventive Effects of Zinc Pretreatment in the Time-course of Cadmium-induced Testicular Toxicity in the Rat (아연 전처리가 시간 경과에 따른 카드뮴 유도 고환 독성에 미치는 보호 효과)

  • Jekal, Seung-Joo;Lee, Kyung-Sun;Chung, Ok-Bong;Im, Hyo-Bin
    • Korean Journal of Clinical Laboratory Science
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    • v.41 no.3
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    • pp.111-122
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    • 2009
  • Cadmium (Cd) is known to exert gonadotoxic and spermiotoxic effects. The present study was performed to investigate the morphological effects and metallothionein (MT) expression by zinc pretreatment in the course of time of cadmium-induced testicular injury in rat. Fifty male Spraque-Dawley rats weighing 160~180 g were divided into two groups : saline-pretreated cadmium group and zinc-pretreated cadmium group. Rats of two groups received subcutaneous injection of saline and 100 mg/kg $ZnSO_4$ at 0, 2, 5 and 8 hrs intervals respectively. Cadmium chloride (4.5 mg/kg $CdCl_2$) was administrated intraperitoneally at 2 hrs after zinc injection and rats were killed 0, 12, 24, 48 and 72 hrs later. Testicular tissue damages, interstitial (Leydig) cells status and MT expression were determined using hematoxylin-eosin stained sections and a computerized image analysis system on sections immunostained with a mouse anti-metallothionein respectively. Zinc pretreatment was significantly reduced testicular damages in five pathological categories after cadmium administation. The number of surviving interstitial cells was significantly higher in the zinc-pretreated group than in the saline-preatreated group at 48 and 72 hrs after cadmium administration. Non-damaged testis showed the positivity of MT staining in spermatogenic cells, Sertoli cells and endothelium of blood vessel, but not in the Leydig cells. The positivity of MT staining in saline-pretreated group was significantly reduced at 24 hrs after cadmium administration, whereas zinc-pretreated group showed strong MT positive staining similar to the 0 hr by 42 hrs after cadmium administration. In damaged testis, MT positive staining was also observed in the Leydig cells of both groups. These results suggest that a major preventive effect of zinc against cadmium-induced testicular toxicity may be due to its ability to reduce the cytotoxicity of cadmium in spermatogenic cells and Leydig cells by inhibiting the susceptibility of the testis to cadmium but not MT production by cadmium.

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Molecular Cloning and Expression of Heat-stable Enterotoxin Gene from Swine Enterotoxigenic Escherichia coli (돼지에서 유래한 병원성 대장균의 내열성 장독소 생산유전자의 Cloning 및 발현)

  • 김교창;도대흥
    • Journal of Food Hygiene and Safety
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    • v.6 no.3
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    • pp.147-155
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    • 1991
  • Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

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Safety and Immunogenicity of Salmonella enterica Serovar Typhimurium llaB in Mice

  • CHO SUN-A;LEE IN-SOO;PARK JONG-HWAN;SEOK SEUNG-HYEOK;LEE HUI-YOUNG;KIM DONG-JAE;BACK MIN-WON;LEE SEOK-HO;HUR SOOK-JIN;BAN SANG-JA;LEE YOO-KYOUNG;PARK JAE-HAK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.609-615
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    • 2005
  • The safety and immunogenicity of an attenuated recombinant Salmonella vaccine strain, Salmonella enterica serovar Typhimurium llaB, was assessed. This vaccine strain could survive in low pH condition, and its ability of intracellular survival did not differ from that of S. enterica serovar Typhimurium UK1, which is the wild-type of the vaccine strain. The mortality of the mice orally administered with the vaccine strain was $50\%$ at the dose of $10^7$ CFU. All mice administered with $10^5\;or\;10^3$ CFU of the vaccine strain survived for 3 days postinoculation (pi). However, all mice administered with more than $10^3$ CFU of the vaccine strain died within 3 days pi. To examine the protective effect of the vaccine strain, mice were orally immunized with $10^4\;and\;10^6$ CFU of the bacteria. Control mice were given with 0.5 ml of phosphate buffered saline (PBS). After 8 days, the mice were challenged with $10^9$ CFU of S. enterica serovar Typhimurium UK1, and mortality was examined for 5 days. The survival rates of the mice immunized with $10^4\;and\;10^6$ CFU of the vaccine strain were $60\%\;and\;80\%$, respectively, whereas all control mice died within 2 days after challenging. To investigate the immunogenicity of S. enterica serovar Typhimurium llaB, mice were orally immunized with $10^5\;or\;10^6$ CFU ml of the vaccine strain. Five mice of each group were sacrificed at 5 and 12 days after immunization, and results showed that immunization of the vaccine strain led to increases of IgG1, IgG2, and IgM titers against S. enterica serovar Typhimurium UK1 in mouse sera, cytokine expressions such as IL-2, IL-4, IL-6, and IL-10 in spleen, and the lymphocyte proliferation response to mitogens (concanavalin A or LPS) stimulation.

Effects in Metabolism and Adipocyte Inflammation Induced by the Complex Herbal Medicine composed of Cornus officinalis, Dioscorea rhizoma, Aurantii fructus, Mori folium in Obese Type 2 Diabetes Mouse Model (산수유 산약 지각 상엽으로 구성된 복합처방투여가 고지방, 고탄수화물 식이로 유발된 비만형 당뇨병 동물모델의 대사인자 및 지방세포 염증반응 조절에 미치는 영향)

  • Han, Su-Ryun;Paik, Sun-Ho;Kwon, Oh-Jun;Ahn, Young-Min;Lee, Byung-Cheol;Ahn, Se-Young
    • The Journal of Korean Medicine
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    • v.33 no.3
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    • pp.184-199
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    • 2012
  • Objectives: There is a steady increase in the prevalence of obesity worldwide and obesity is often accompanied by inflammation. Although much emphasis has been placed on the adipose tissue inflammation in obesity, a study with herbal medicine is few. This study aimed to investigate the antidiabetic and anti-inflammatory effect of a complex herbal medicine (CHM) composed of Cornus officinalis, Dioscorea rhizoma, Aurantii fructus, and Mori Foliumon on obese type 2 diabetes mice. Methods: Type 2 diabetes mellitus and obesity were induced by Surwit's high fat, high sucrose diet for 8 weeks. Mice were divided into ND (normal diet, n=10), HFD (high fat and high sucrose diet, n=10), CHM (high fat and high sucrose diet with complex herbal medicine, n=10) and Met (high fat and high sucrose diet with metformin, n=10) groups. The body weight, fructosamine and OGTT (oral glucose tolerance test) were measured. After 8 weeks the blood samples of all mice were taken from the heart, and lipid profiles were measured. Epididymal fat pad, histological size of the adipocyte tissue and liver weights were measured. Inflammatory markers such as leptin and adipocyte tissue macrophage were measured to evaluate the effect of CHM on adipocyte tissue inflammation. Results: Compared with the HFD group, there was an improvement in OGTT and epididymal fat decreased in the CHM group. White adipocyte size and adipocyte tissue macrophage decreased in CHM group. Conclusions: These results suggest that CHM has antidiabetic and anti-inflammatory effects in high fat, high sucrose diet induced obese mice.

Natural Antioxidant Activity of Ethanol Extracted from Bovine Bile ; Biological Effects and Characterization (초식동물 쓸개즙 추출물의 천연항산화 성분; 생물학적인 기능 및 특성규명)

  • Shim, Jae-Han;Park, Myung-Woo;Lim, Kye-Taek
    • Korean Journal of Environmental Agriculture
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    • v.18 no.3
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    • pp.221-228
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    • 1999
  • This study was carried out to extract the natural antioxidants from Bovine bile and to investigate their effects on various antioxidant activities. It also characterized the patterns of antioxidants by GC/FID and GC/MS. The antioxidative activities and chemical structure of the antioxidant were elucidated by examining the effects of biological activity and the analysis of GC/MS. The antioxidant materials extracted from bovine bile were isolated and purified by silica gel column chromatography and TLC. It was confirmed that there were effects of antioxidants such as Xanthine Oxidase(XO) and Glutathione-S-Transferase(GSH-T) on antioxidative activities. When they were compared with BHT, bile extracts showed the relative effects of 51.2% on the antioxidant activity, the inhibition effects of 48.3% on XO activity, and the synergism effects of 85.7% on the GSH-T activity. According to the results of investigation at neuron cell of mouse, the rate of cell activity in the treatment of 6mM glutathione was 96%, While it in the treatment of 140mg of bile extract was 78%. Based on the TLC analysis of EtOAc extracts from the Bovine bile, the antioxidant activity appeared at $R_f$ value, 0.72. These results suggested that the antioxidant may be coprostan 3-ol.

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Suppressive Effects of Lees from Sweet Potato Soju on LPS-induced Inflammatory Responses in RAW 264.7 Cells (고구마 소주 주박에 의한 RAW 264.7 세포주에서 lipopolysaccharide로 유도된 염증 반응의 억제 효과)

  • Lee, Seung-Hoon;Kwon, Min-Jeong;Kim, Soon Young;Sohn, Ho-Yong;Shin, Woo-Chang;Kim, Jong-Sik
    • Journal of Life Science
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    • v.26 no.1
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    • pp.117-122
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    • 2016
  • In the current study, the ethanol extracts and their subsequent organic solvent fractions from lees of sweet potato soju were prepared and the prepared samples were designated as from KSD-E8-1 to KSD-E8-5. Their effects on cell viability and nitric oxide (NO) production in mouse macrophage RAW 264.7 cells were investigated. The results showed that the ethyl acetate fraction (KSD-E8-3) of lees extracts from sweet potato soju significantly decreased nitric oxide (NO) production in LPS-activated RAW 264.7 cells, whereas they did not affect cell viabilities. The fraction KSD-E8-3 reduced the expression of pro-inflammatory genes such as COX-2, iNOS and TNF-alpha and also decreased protein expression of iNOS in a dose dependent manner, which were detected with RT-PCR and Western blot analysis, respectively. In addition, we detected the expression of mitogen-activated protein kinases (MAPKs) such as p38, JNK, and ERK1/2 and their phosphorylated forms. The results indicated that the treatment of the fraction KSD-E8-3 did inhibit phosphorylation of p38, JNK, and ERK1/2 MAPKs, indicating that the fraction KSD-E8-3 regulates LPS-induced inflammatory response via suppressing MAPK signaling pathway. Overall, these results may contribute to understand the molecular mechanism of anti-inflammatory effects by the ethyl acetate fraction of lees extracts from sweet potato soju.

Molecular Cloning and Characterization of Bovine CYP26A1 Promoter (소 CYP26A1 유전자 프로모터의 molecular cloning 및 특성)

  • Kwak, Inseok
    • Journal of Life Science
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    • v.26 no.1
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    • pp.42-49
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    • 2016
  • The retinoic acid (RA) plays an important role in the growth and development of many cells, and bioactive RA concentration is regulated by several enzymes, including CYP26A1. The expression of the CYP26A1 gene is regulated by RA, and the CYP26A1 gene is one of the candidates for RA-responsive genes. Although CYP26A1 genes are cloned from several animals, cloning of the CYP26A1 gene from cows has not been reported yet. The promoter region of CYP26A1 from cows was cloned by PCR and analyzed by sequence alignment with human and mouse CYP26A1. The RA-responsive element (RARE), DR-5 (ttggg), was located in this region and was perfectly conserved. The promoter region of bovine CYP26A1, which contains DR-5, was ligated to the luciferase reporter gene on transient transfection assays. The expression of CYP26A1-Luc promoter was activated by ATRA treatment in lung-derived mtCC cells. Co-transfection with RAR-α or -β with ATRA significantly activates the expression of CYP26A1-Luc promoter; however, it was less effective with either RAR-γ or RXR-γ. In addition, the endogenous gene expressions measured by Q-RT-PCR in mtCC cells were not significantly affected by ATRA treatment for 2 days; however, the expression of the endogenous CYP26A1 gene was diminished sharply at day 3 with ATRA treatment. In conclusion, the promoter region of bovine CYP26A1 contains conserved DR-5 RARE, which functions as a binding site for RAR-α or -β, and it is involved in the regulation of CYP26A1 gene expression and the control of RA signaling in mtCC cells.

Amyloid-β Levels in Mice Hippocampus According to the ALDH2 Enzyme Activity followed Ethanol Exposure for 8-Weeks (ALDH2 효소 활성과 8주간 에탄올 노출에 따른 해마조직의 아밀로이드 베타 발현)

  • Moon, Sun-In;Eom, Sang-Yong;Yim, Dong-Hyuk;Song, Sun-Ho;Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1636-1640
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    • 2011
  • Alzheimer's disease (AD) is a progressive neurodegenerative disease, resulting in the loss of cognitive function. Mitochondrial aldehyde dehydrogenase (ALDH2) has been proposed to be a risk factor for the development of AD, but there is still controversy about that. In this study, we demonstrated the role of ALDH2 enzyme activity on amyloid-beta (A${\beta}$) and nuclear factor kappa B (NF-${\kappa}B$) expression in mice brain following ethanol exposure for 8 weeks. Five male Aldh2 (+/+) and Aldh2 (-/-) mice, 8 weeks-old of age (C57BL/6J strain), in each group were exposed to ethanol for 8 weeks (2 g/kg wt./day) using gavage. Those in the control groups received 0.9% saline alone. Results showed a difference in expression level of A${\beta}$ in the hippocampus after ethanol exposure according to the ALDH2 enzyme activity (p<0.05), but not in the level of NF-${\kappa}B$). Our results suggest a possibility that ALDH2 enzyme activity may be an important role in the development of AD.

Studies of Inhibitory Mechanism on Melanogenesis by Partially Purified Asiasari radix in α-MSH Stimulated B16F10 Melanoma Cells (세신추출물이 α-MSH 자극에 의한 B16F10 세포의 멜라닌생성에 미치는 영향)

  • Jang, Ji-Yeon;Kim, Ha-Neui;Kim, Yu-Ri;Kim, Byung-Woo;Choi, Yung-Hyun;Choi, Byung-Tae
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1617-1624
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    • 2010
  • Recently, it has been found that Asiasari radix showed a hypopigmenting effect on melanogenesis through activation of mitogen-activated protein kinase (MEK)/extracellular signal-activated kinase (ERK) in B16F10 melanoma cells. However, the hypopigmenting effect of A. radix on the $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH)-stimulated melanogenesis has remained unknown. The purpose of this study was to investigate the inhibitory mechanism of the partially purified A. radix (PPAR)-induced hypopigmentating effects on $\alpha$-MSH-stimulated melanogenesis in B16F10 mouse melanoma cells. PPAR strongly inhibited tyrosinase activity and leads to decreased melanin synthesis in $\alpha$-MSH-stimulated B16F10 melanoma cells. PPAR also decreased the $\alpha$-MSH-induced over-expression of the melanogenic enzymes, tyrosinase, tyrosinase-related protein (TRP)-1, dopachrome tautomerase (Dct) and microphthalmia-associated transcription factor (MITF). We further showed that PPAR inhibits $\alpha$-MSH-induced melanogenesis via phosphorylation of MEK/ERK and PI3K/Akt, and that their activation was blocked by MEK inhibitors, PD98059 and PI3K inhibitors, LY294002 in $\alpha$-MSH-stimulated B16F10 melanoma cells. These results suggest that PPAR inhibits $\alpha$-MSH-induced melanogenesis by activation of MEK/ERK and PI3K/Akt through MITF degradation, which may lead to down-regulation of tyrosinase.