• Title/Summary/Keyword: inhibition ELISA

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Adenine, new anti-wrinkle agent.

  • Kim, Y. J.;Kim, Y. S.;S.Y. Eom;Kim, J. H.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.804-819
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    • 2003
  • It has been known that adenine is a very important material in living cells. Because, adenine is a member of nucleotide base, so it takes part in DNA, RNA and ATP synthesis. There are many reports that adenine participated in ingredients, especially DNA, RNA, NADH and ATP, affect on the cell. As well adenosine, conjugated adenine to glycoside, was known to anti-wrinkle compound. But there is no report whether adenine shows a good effect on the skin, especially anti-wrinkle. So, in this study, we tested whether adenine affects cell proliferation, collagen synthesis, collagenase synthesis inhibition in human dermal fibroblasts. In addition, we performed clinical study with adenine cream. Cell proliferation effect was tested by MTT assay. Collagen and collagenase synthesis were measured by Immunoassay with ELISA kit. Clinical study was performed by IECK according to KFDA Functional Cosmetic method. The results of cell proliferation show that 10$^{-6}$ ~10$^{-8}$ % of adenine increases cell proliferation about 50 % compare with non-treated control. At 10$^{-7}$ ~10$^{-10}$ %, adenine increases type I collagen synthesis about 50%, decreases type I collagenase about 22% compare with non-treated control. The results of clinical study show that 0.05% adenine treated group reduces wrinkle significantly compare with placebo treated group. Therefore adenine may be a new anti-wrinkle candidate, through increases cell proliferation and collagen synthesis dramatically. And it decreases collagenase synthesis. So adenine could be used as a new anti-wrinkle agent.

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Acetylcholinesterase Inhibitory and Antioxidant Properties of Aster yomena Extract (쑥부쟁이 추출물의 아세틸콜린에스테라제 저해 및 항산화 활성)

  • Bae, Jong-Sup;Kim, Tae-Hoon
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.121-126
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    • 2009
  • Objectives : To evaluate the radical scavenging and acetylcholinesterase (AChE) inhibitory activities of the ethylacetate (EtOAc)-soluble portion of a methanolic extract of Aster yomena, three different assay systems were performed. Methods : The antioxidant activity of A. yomena extract was tested as its capacity to scavenging free radicals of DPPH and $ABTS^+$, which has been widely used to evaluate the antioxidant activity of natural products from plant sources. AChE inhibitory activity was tested against mouse brain AChE by spectrophotometric method of Ellman using ELISA microplate reader. Results : The methanolic extract of A. yomena was fractionated and the EtOAc-soluble portion showed significant AChE inhibitory and free radical scavenging effects. Also the EtOAc-soluble portion revealed the highest phenolic contents as compared to the other extracts. Conclusions : These results indicate that phenolic compounds may be important constituents that give rise to the anti-AChE and antioxidative activities of A. yomena extract. Further phytochemical studies on this plant, for nutraceutical or pharmaceutical application, are warranted.

Anti-Inflammatory Effects of Hydroethanolic Extract from Ehretia asperula on Lipopolysaccharide-Stimulated RAW264.7 Macrophages

  • Bao Le;Vo Thi Kim Hong;Seung Hwan Yang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1340-1347
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    • 2024
  • Ehretia asperula is a medicinal plant of the Ehretiaceae family used to treat inflammatory disorders, but the underlying mechanisms are not fully elucidated. The anti-inflammatory potential was determined based on enzyme cyclooxygenase-2 (COX-2) inhibition, which showed that the 95% ethanol extract (95ECH) was most effective with a half-maximal inhibitory concentration (IC50) value of 34.09 ㎍/mL. The effects of 95ECH on phagocytosis, NO production, gene, and protein expression of the cyclooxygenase 2/prostaglandin E2 (COX-2/PGE2) and inducible nitric oxide synthase/ nitric oxide (iNOS/NO) pathways in lipopolysaccharide (LPS)-induced RAW264.7 cells were examined using the neutral red uptake and Griess assays, reverse-transcriptase polymerase chain reactions (RT-PCR), and enzyme-linked immunosorbent assays (ELISA). The results showed that 95ECH suppressed phagocytosis and the NO production in activated macrophage cells (p < 0.01). Conversely, 95ECH regulated the expression levels of mRNAs for cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) as well as the corresponding proteins. In addition, PGE2 production was inhibited in a dose-dependent manner by 95ECH, and the expression of iNOS and COX-2 mRNAs was decreased in activated macrophage cells, as expected. Therefore, 95ECH from E. asperula leaves contains potentially valuable compounds for use in inflammation management.

Inhibitory Effect on Infection of Plant Viruses by Filtrate Powder from Culture Broth of Acinetobacter sp. and Its Mode of Action (Acinetobacter sp. 배양여과액 분말제제의 식물바이러스에 대한 감염억제 효과 및 작용)

  • Kim Mi-Soon;Jung Min-Young;Kim Yun-Sung;Jang Cheol;Hwang In-Cheon;Ryu Ki-Hyun;Choi Jang-Kyung
    • Research in Plant Disease
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    • v.12 no.2
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    • pp.91-98
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    • 2006
  • A filtrate powder, designated as KNF2022, produced from culture broth of Acinetobacter sp. KTB3 was tested for their inhibitory effects on Pepper mild mottle virus (PMMoV) infection to Nicotiana glutinosa or N. tabacum cv. Xanthi nc. When 1/100 dilution with distilled water was treated to the plants and PMMoV was inoculated, the inhibition was estimated to be 94.3 and 95.6%, respectively. The same concentrations of KNF2022 inhibited infections of Pepper mottle virus (PepMoV) and Cucumber mosaic virus (CMV) on Chenopodium amaranticolor by 97.1 and 92.5%, respectively. Duration of inhibitory activity of the filtrate powder from Acinetobacter sp. culture broth against PMMoV infection on N. glutinosa was maintained for 2 days at 80% inhibition level, however, the inhibitory effect was diminished from 4 days after treatment to 50% levels. To evaluate inhibitory effects on systemic host plants of the antiviral agent, symptom developments of PMMoV, PepMoV and CMV on KNF2022-treated pepper plants were investigated. Delayed symptom developments until 10 days after inoculation (DAI) were observed for all the three viruses when the viruses were inoculated individually, and these delayed symptom development effects were maintained until 30 DAI in case of PepMoV. Moreover, PepMoV was not detected by RT-PCR and ELISA until 30 DAI. These delayed symptom development effects were diminished in all combinations of three virus co-inoculations due to synergism of three viruses on symptom developments. Inhibitory effect of KNF2022 was verified under electron microscopic examinations using purified virus preparations. Particles of PMMoV and PepMoV were observed on specimens from 5 min after KNF2022 treatment, and the particle sizes were reached in the range of 200-250 nm and 400-600 nm, respectively. Furthermore, the viral particles were destructed and particle sizes were reached in the range of 100-150 nm and 300-500 nm, respectively, on 60 min after treatments. Reduction of local lesion numbers on N. tabacum cv. Xanthi nc and C. amaranticolor were accompanied with reduction of virus particle sizes. In the case of CMV destructed particle numbers were also increased according to incubation period after KNF2022 treatment and local lesions on C. amaranticolor were reduced.

A Study of matrix metalloproteinase-9 inhibitor in root bark of ulmus davidiana planchon (유근피내의 Matrix Metalloproteinase-9 활성 억제제에 관한 연구)

  • Kong, Kwang-Hoon;Han, Kee-Jung;Lee, Kwang-Soo;Cho, Sung-Hye
    • Analytical Science and Technology
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    • v.18 no.2
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    • pp.104-111
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    • 2005
  • Several solvents were used to fractionate an extract obtained from the chapped root bark of Ulmus davidiana Planchon. The each fractionary part was condensed under reduced pressure and then examined to investigate the inhibitory effect on MMPs by modified gelatin zymography, where EA fraction showed the inhibition effect on the activity of MMPs. A compound showing inhibition effect on the MMPs was isolated and purified from EA fraction. Under IR, $^1H$- and $^{13}C$- NMR analyses it is very close to a catethin. This substance showed 48% inhibition effect on measurement of MMP-9 activity at 5 mM and 43% at 10 mM. To verify the effect of this substance on cells, human hepatoma, SK-Hep-1 cells as a cancer model, and Chang liver cells as a normal model were selected. MTT assay was performed to examine the cell viability by treatment of $1{\mu}L/mL$ of the purified substance on cells. The purified substance showed negligible toxicity on human liver cell line.

The Efficacy of Saururus chinensis on Cervical Cancer Cells : The Inhibitory Effect on the Function of E6 and E7 Oncogenes of HPV Type 16 (삼백초 추출물의 자궁경부암세포 억제 효능)

  • Chung, Yeon-Gu;Lee, Hae-Sook;Lee, Kyung-Ae;Joung, Ok;Oh, Won-Keun;Kim, Kwang-Dong;Lim, Jong-Seok;Moon, Ja-Young;Cho, Yong-Kweon;Park, Sue-Nie;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.46 no.6
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    • pp.426-432
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    • 2002
  • Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV type 16 and 18) and cervical cancer has been well known. An extract of Saururus chinensis, named as PE-46, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. PE-46 inhibited the proliferation of human cervical cancer cell lines in a dose response manner. PE-46 also inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, PE-46 inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by PE-46. SiHa cells treated with PE-46 induced G0/G1 arrest, resulting in inhibition of growth. Our study showed that the PE-46 can inhibit the cervical carcinomas via both inhibition of bindings between oncogenes and tumor suppressors, and inhibition of G1longrightarrowS transition. PE-46 inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus could be used as a putative modulating agent for the treatment of cervical carcinomas caused by HPV.

Anti-wrinkle Compounds Isolated from the Seeds of Arctium lappa L. (우방자에서 분리한 주름개선 화합물)

  • Hwang, Ju-Young;Park, Tae-Soon;Kim, Dong-Hee;Hwang, Eun-Young;Lee, Jung-Noh;Lee, Ji-Young;Lee, Ghang-Tai;Lee, Kun-Kook;Son, Jun-Ho
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1092-1098
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    • 2012
  • This study was carried out to discover the skin wrinkle reducing components in the seeds of Arctium lappa. The isolation of a methylene chloride-soluble fraction of 70% ethanol extract from the seeds of Arctium lappa using a procollagen type-1 synthesis and MMP-1 activity resulted in the isolation and identification of four lignin compounds: arctiin, arctigenin, matairesinol, and diarctigenin. All structures were confirmed via NMR and MS spectroscopic data. To determine cell viability and procollagen type-1 synthesis, human dermal fibroblasts were treated with 10-100 ${\mu}M$. As a result, none of the four compounds showed cytotoxicity up to 50 ${\mu}M$. We also investigated their procollagen type-1 synthesis and MMP-1 inhibition activity and found that arctiin had the highest activity in terms of both procollagen synthesis and MMP-1 inhibition among all four compounds. Putting all the data together, we suggest that arctiin be used in cosmetics as an anti-wrinkle material.

Roots of Daucus carota sativa abrogates acute phase of Inflammation by the Inhibition of NO and Pro-Inflammatory Cytokine Production (NO와 Pro-Inflammatory Cytokine의 억제를 통한 호라복(胡蘿蔔)의 항염증효과)

  • Lee, Dong-Jin;Park, Sang-Mi;Hwangbo, Min;Jung, Tae-Young;Kim, Sang-Chan;Jee, Seon-Young
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.2
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    • pp.45-57
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    • 2013
  • Objectives : Daucus carota sativa has been frequently used as food supplements in many of the Asian countries, and a nutritional medical drug in traditional medicine. This research investigated the effects of Daucus carota sativa extract (DCE) on acute phases of inflammation in Raw 264.7 cells treated with lipopolysaccharide (LPS) in terms of the inhibition of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines production. Methods : NO, $PGE_2$, tumor necrosis factor (TNF)-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6 contents were assayed by ELISA, and expressions of inflammation-related proteins such as inducible NO synthase (iNOS) were determined by immunoblot analyses. Results : DCE treatment attenuated the LPS ability to increase the productions of NO and $PGE_2$ as well as the protein level of iNOS in a concentration-dependent manner. Consistently, treatment of the cells with DCE suppressed the production of TNF-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6. DCE also caused decreases of inhibitor of ${\kappa}B{\alpha}$ phosphorylation induced by LPS in the cells, which means DCE inhibition of NF-${\kappa}B$ activity. Furthermore, DCE blocked LPS-induced phosphorylation of p38 and SAPK/JNK. Conclusion : This study showing here may be of help to understand the action mechanism of DCE, and provide the information for the medical use of Daucus carota sativa for the inflammatory disease.

Effect of CpG Oligodeoxynucleotides on Airways of Mice with Established Airways Inflammation (기도 염증이 유발된 생쥐에서 CpG Oligodeoxynucleotides가 미치는 효과)

  • Hwang, Hei-Won;Kim, Su-Jin;Kim, Won-Duk;Cho, Sung-Min;Lee, Dong-Suk;Choi, Sung-Min
    • Clinical and Experimental Pediatrics
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    • v.45 no.7
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    • pp.875-883
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    • 2002
  • Purpose : Airways eosinophilia and increased IgE, characteristic features of asthma, result from a predominant Th2 response. In this study, we investigated the effect of CpG oligodeoxynucleotides (ODNs) on the inhibition of airways eosinophilia in mice with established airway inflammation. We also investigated the immunological mechanisms involved. Methods : Groups of BALB/c mice were sensitized intradermally with ovalbumin(OVA). At week 10, airway inflammation was induced by intranasal challenge of the mice with OVA. At week 14, the mice were challenged intranasally again with OVA in the presence and without the presence of CpG ODNs. Mice with saline administration served as negative controls. Bronchoalveolar lavage fluids(BALF) were obtained and eosinophils were counted. Th1 and Th2 cytokines in the spleen cell cultures were measured by ELISA. Serum OVA-specific IgE and IgG2a antibodies were also measured by ELISA. Results : BALF eosinophils were significantly inhibited in the CpG ODNs-treated mice(P<0.01). IgE and IgG2a levels increased significantly in both CpG ODNs-treated and untreated groups as compared to the negative control group; there was, however, no significant difference between the two groups four days after intranasal administration of CpG ODNs. Cytokine analysis revealed decreased production of IL-4, IL-5, and IL-13 and increased production of IL-12 in the CpG ODNs-treated group as compared to the untreated group. Interestingly, $IFN-{\gamma}$ levels were not upregulated in the CpG ODNs-treated group. Conclusion : CpG ODNs vaccination is a potentially useful approach for reversing airways eosinophilia in mice with established airways inflammation.

Studies of the egg drop laying diseases from the mass zone layer (산란계 밀집지역의 산란저하성 질병에 관한 연구)

  • Lee Jeoung-Won;Eum Sung-Shim;Park In-Gyu;Bea Joung-Jun;Joung Dong-Suk;Song Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.28 no.2
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    • pp.121-146
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    • 2005
  • Newcastle disease (ND), infectious bronchitis (IB), low pathogenic avian Influenza (LPAI) and fowl typhoid (FT) have been known as egg drop laying diseases because of the serious layer damage from mass zone layer. In this study, such egg drop laying diseases were investigated. To access this study, we peformed to evaluate antibody titers in serum and isolated bacteria and virus from organs and feces on May, July and September in 2003. The distribution of ND from January to May, IB and LPAI from October to February of the next year, and FT from March to September were inspected by the question survey in 21 farms. ND revealed to be positive rates of 490 to 474 $(96.7\%)$ in May, 510 to 506 $(99.2\%)$ in July and 510 to 510 $(100\%)$ in September with hemagglutination inhibition (HI) test. The mean antibody titers were 10.2, 9.9 and 10.2, respectively. With regard to IB, 484 out of 490 samples $(98.7\%)$ in May, 508 of 510 $(99.6\%)$ in July and 509 of 510 $(99.8\%)$ in September showed positive results and the mean antibody titers were gradually increased with 8.2, 9.0 and 9.4, respectively. According to HI test of LPAI, the positive results were shown in 442 of 480 $(92.1\%)$, 394 of 494 $(79.8\%)$ and 402 of 483 $(83.2\%)$ in May, July and September, respectively The mean antibody titers were decreased with 4.6, 4.3 and 4.0. The distribution of LPAI also elicited the positive rates of 480 to 475 $(99.0\%)$ in May, 494 to 485$(98.2\%)$ in July, 483 to 472 $(97.7\%)$ in September as determined by ELISA and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. Compared ELISA results with HI test of LPAI the positive results were 480 to 422 $(92.1\%),\;475(99.0\%),\;494\;to\;394 (79.8\%),\;485 (98.2\%)\;and\;483\;to\;402(83.2\%),\;472(97.7\%)$. Therefore, the positive rate determined by ELISA was higher than that of HI test with 6.9, 18.4 and $14.5\%$, respectively. When performed RT-PCR for ND using organ and feces samples, the pathotypes were detected $5(15.6\%)\;in\;May,\; 2(5.3\%) in\;July,\;2(7.1\%)$ in September but there is no samples showing positive band for LPAI. In attempt to isolate Salmonella gallinarum, bacteria were obtained from 4 cases (12.5%) in May, 9 (23.6%) in July, 5 (17.8%) in September. Thus the highest rate for isolation revealed to be shown in July When evaluated the antimicrobial susceptibility to 18 isolated strains of 5. gallinarum, bacteria were sensitive to trimethoprim/sulfamethox$(61.1\%),\;kanamycin\;(55.5\%),\;ampicillin\;(55.5\%)$ and amoxacillin/clavulanic acid $(55.5\%)$, cephalothin $(50.0\%)$, but resistant to penicillin $(88.9\%)$, streptomycin $(88.9\%)$, erythromycin $(83_4\%)$ and tetracycline $(61.1\%)$. According to HI test of ND and LPAI using captured 164 wild Korean tree sparrows (Passer nontanus), the positive rates were $47.6\%\;and\;57.3\%$, and the mean HI titers were 5.32 and 4.02, respectively. 71 $(43.2\%)\;and\;58(35.3\%)$ in captured sparrows also showed more than 4 titers for HI test to ND and LPAI, respectively However, the attempt for isolation of viruses failed in all samples.