• Title/Summary/Keyword: ELISA.

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The Effects of Rhus Extracts on The Cytotoxicity on Cancer Cells and E6 and E7 Oncogenes of Human Papillomavirus Type 16 (옻 추출물의 세포독성 및 자궁 경부암 바이러스 암 유발인자 E6 와 E7의 작용에 미치는 효과)

  • Cho, Young-Sik;Joung, Ok;Cho, Cheong-Weon;Lee, Kyung-Ae;Shim, Jung-Hyun;Kim, Kwang-Soo;Lee, Hong-Soo;Seung, Ki-Seung;Yoon, Do-Young
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1389-1395
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    • 2000
  • Cervical cancer has been one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papillomaviruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, Rhus, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Rhus inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, Caski) and HaCaT keratinocytes in a dose response manner. In vitro binding assay and ELISA showed that Rhus inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Rhus inhibited the in vitro binding of E7 and Rb which essential tumor suppressor for the control of cell cycle. The level of mRNA for E6 was also decreased by Rhus while that of E7 mRNA was not changed. Our data suggested that Rhus inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas by HPV.

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TISSUE LEVELS OF INTERLEUKIN-1α, INTERLEUKIN-1β AND TUMOR NECROSIS FACTOR-α IN PULPAL AND PERIAPICAL PATHOSIS (치수 및 치근단병소에서 interleukin-1α, interleukin-1β, tumor necrosis factor-α의 분포에 관한 연구)

  • Ko, Hyun-Jung;Chung, Kwan-Hee;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.23 no.1
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    • pp.316-327
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    • 1998
  • This study was designed to examine the tissue levels of interleukin-$1{\alpha}$(IL-$1{\alpha}$), interleukin-$1{\beta}$(IL-$1{\beta}$) and tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) in inflamed human dental pulps and periapical lesions, and to determine the relationship between each cytokine and pulpal and periapical pathosis. The pulps used in this experiment, were obtained in routine endodontic treatment and the periapical lesions in periapical surgery after clinical diagnoses were performed. These specimens were divided into four groups as normal pulp group(control group, n=9), acute pulpitis group(n=g), chronic pulpitis group(n= 10) and periapical lesion group(n= 18) and stored in liquid N2. For extract preparation, tissues were finely minced with a scalpel, and the fragments were incubated in $0.5m\ell$ homogenizing buffer (0.1 mol/L potassium chloride, 0.02 mol/L TRIS; pH=7.6) for two hours and grinded with glass homogenizer. Debris was removed by centrifugation and supernatants were immediately tested with enzymelinked immunosorbent assay (ELISA, R&D Co., Minneapolis, USA). Following results were obtained; 1. The concentrations of IL-$1{\alpha}$ in all experimental groups were significantly higher than in control group(p<0.05). And the concentrations of IL-$1{\alpha}$ in periapical lesion group were somewhat higher than in two pulpitis groups, but the differences among those groups were not stastically significant (p>0.05). 2. The concentrations of IL-$1{\beta}$ in all experimental groups were significantly higher than in control group (p<0.05), and all the experimental groups expressed similar concentrations. 3. The concentrations of TNF-${\alpha}$ in all experimental groups were higher than in control group but only the differences between chronic pulpitis group and control group were statistically significant(p<0.05). And the concentrations of TNF-${\alpha}$ in acute and chronic pulpit is groups were higher than in periapical lesion group but only the differences between chronic pulpitis group and periapical lesion group were statistically significant (p<0.05). 4. There was significant correlation only between IL-$1{\alpha}$ and IL-$1{\beta}$ in periapical lesion group (p<0.05).

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Study on the Production and Management of Aquatic Animal : Application of ELISPOT-Assay for the Detection of Antibody Secreting Cells in Flounder, Paralichthys olivaceus (수산생물의 생산과 관리에 관한 기초연구 : ELISPOT 기법을 이용한 넙치의 항체생성 세포분석)

  • HA Jai Yi;PARK Jun-Hyo;KIM Myoung Sug;CHUNG Joon-Ki;JEONG Hyun Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.4
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    • pp.420-426
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    • 1999
  • We examined the immune response in flounder, Paralichthys olivaceus, with immunization of formalin killed Edwardsiella tarda as an antigen. The ELISPOT-assay (enzyme-linked immunospot assay) was optimized technically and applied to count the number of total and specific antibody secreting cells (TASC and SASC) in lymphocytes of different lymphatic organs. Incubation of lymphocytes on 96 well plate for more than 2.5hrs came out enough time in ELISPOT-assay for counting the antibody secreting cells in the anterior kidney and spleen. However, too much of plate-coated antigen or rabbit anti-flounder immunoglobulin for SASC or TASC counting, respectively, was appeared to decrease the sensitivity of the assay system. Specificity of the system was also confirmed by the absence of TASC in lymphocytes treated with cycloheximide to prevent protein synthesis. The peak numbers of SASC appeared at wk 3 post immunization after that there was a sharp decrease and reached to almost zero at wk 7. In the spleen and kidney, the timing and numbers of SASC on peak response were concurrent without preferential organ distribution. The specific antibody level in the sera increased rapidly between wk 2 and 3 after immunization, i.e. like the specific cellular response found with ELISPOT-assay on that period, However, the remained high level of specific serum antibody from wk 5 after immunization until the end of experiment was clearly distinguishable from the kinetics of SASC response decreased sharply.

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Effects of Heat Treatment on the Antigenicity of Gamma-Irradiated Egg White Albumin (감마선 조사된 난백 알부민의 항원성에 미치는 가열의 영향)

  • 이주운;육홍선;조경환;김미리;김천제
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.848-853
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    • 2001
  • This research was conducted to study the effects on antigenicities (allergenicity) and structural changes of gamma-irradiated hen’s egg albumin (ovalbumin, OVA) by heating. Three groups of OVA solution (2.0 mg/mL) were prepared; 1) heat treatment; 2) irradiation after heating; 3) heating after irradiation. Samples were isothermally heated and/or irradiated at the absorption dose of 10 kGy. Competitive indirect ELISA was individually formatted with egg-allergic patients IgE (P-IgE), and mouse murine monoclonal IgG (M-IgG) and rabbit polyclonal IgG (R-IgG) for evaluating bindinhg abilities of antibodies to OVA in the sample solutions. Binding abilities of antibodies to thermally denatured OVA were changed : R-IgG to the sample treated with above 6$0^{\circ}C$, M-IgG to that above 7$0^{\circ}C$, and P-IgE to that above 8$0^{\circ}C$, respectively. P-IgE did not well recognize OVA heated at 8$0^{\circ}C$ and the above. However, binding abilities of M-IgG and R-IgG highly in creased. Significant differences of binding abilities were not observed in all samples with the combination of heat treatment and irradiation, regardless the order of the treatment. Turbidity of samples in creased both by heating and by irradiation, and the increase by irradiation was much higher than by heating. These results showed that allergenicity of OVA reduced by gamma irradiation was not affected by heating.

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Effects of 17β-Estradiol on Colonic Permeability and Inflammation in an Azoxymethane/Dextran Sulfate Sodium-Induced Colitis Mouse Model

  • Song, Chin-Hee;Kim, Nayoung;Sohn, Sung Hwa;Lee, Sun Min;Nam, Ryoung Hee;Na, Hee Young;Lee, Dong Ho;Surh, Young-Joon
    • Gut and Liver
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    • v.12 no.6
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    • pp.682-693
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    • 2018
  • Background/Aims: Intestinal barrier dysfunction is a hallmark of inflammatory bowel diseases (IBDs) such as ulcerative colitis. This dysfunction is caused by increased permeability and the loss of tight junctions in intestinal epithelial cells. The aim of this study was to investigate whether estradiol treatment reduces colonic permeability, tight junction disruption, and inflammation in an azoxymethane (AOM)/dextran sodium sulfate (DSS) colon cancer mouse model. Methods: The effects of $17{\beta}$-estradiol (E2) were evaluated in ICR male mice 4 weeks after AOM/DSS treatment. Histological damage was scored by hematoxylin and eosin staining and the levels of the colonic mucosal cytokine myeloperoxidase (MPO) were assessed by enzyme-linked immunosorbent assay (ELISA). To evaluate the effects of E2 on intestinal permeability, tight junctions, and inflammation, we performed quantitative real-time polymerase chain reaction and Western blot analysis. Furthermore, the expression levels of mucin 2 (MUC2) and mucin 4 (MUC4) were measured as target genes for intestinal permeability, whereas zonula occludens 1 (ZO-1), occludin (OCLN), and claudin 4 (CLDN4) served as target genes for the tight junctions. Results: The colitis-mediated induced damage score and MPO activity were reduced by E2 treatment (p<0.05). In addition, the mRNA expression levels of intestinal barrier-related molecules (i.e., MUC2, ZO-1, OCLN, and CLDN4) were decreased by AOM/DSS-treatment; furthermore, this inhibition was rescued by E2 supplementation. The mRNA and protein expression of inflammation-related genes (i.e., KLF4, NF-${\kappa}B$, iNOS, and COX-2) was increased by AOM/DSS-treatment and ameliorated by E2. Conclusions: E2 acts through the estrogen receptor ${\beta}$ signaling pathway to elicit anti-inflammatory effects on intestinal barrier by inducing the expression of MUC2 and tight junction molecules and inhibiting pro-inflammatory cytokines.

Current Status and Developments of Comparative Education at Universities in Korea and World Wide (세계적 동향에 비추어 본 우리나라 대학의 비교교육학 교육현황과 과제)

  • Hyun, Kyung S.;Park, Elisa L.
    • Korean Journal of Comparative Education
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    • v.18 no.4
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    • pp.1-26
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    • 2008
  • The purpose of this study is to present the overall landscape of Comparative Education study in teacher training program at universities in Korea. The Comparative Education in Korea is relatively young without a lengthy tail on its name and still on the way of upgrading itself, however various efforts are gathering for the development of Comparative Education. With this understanding, this paper diagnoses the current status of Comparative Education in Korea by analyzing 1) the number of institutions which provide Comparative Education lectures for their teacher training program, 2) aims of Comparative Education teaching, 3) curricular and course titles, 4) teaching methods and textbooks, and 5) current issues and problems currently laid on Comparative Education in Korea designated by actual lecturers and professors who are in charge of Comparative Education lectures. The historical development of Comparative Education in Korea will be briefly presented at the beginning part of the chapter. To gauge the comparative position of Korean Comparative Education, the second half of this paper introduces the cases of Comparative Education at universities worldwide, presenting various countries' status of Comparative Education along with their developmental status. As the first research which pays an attention on the current figure of Comparative Education in Korea, it will form an impetus on why we should teach college students about comparative education and on what next step should be carried for further development of Comparative Education at universities based on the comparative understanding of other countries. Since it is in the transitional stage in terms of its development, it would be critical to have a self-analysis of the current status, which will help to lay the comprehensive understanding background for designating the future direction of its development.

Antimetastatic Effects of Crude Polysaccharide Isolated from Polygonati Rhizoma on 4T1 Breast Cancer Cells by Activation of Innate Immune System (황정(黃精)으로부터 유래한 조다당류의 선천면역 활성에 의한 유방암 세포주 전이 억제 효과)

  • Ji, Hae-Ri;Hwang, Deok-Sang;Lee, Chang-Hoon;Jang, Jun-Bok;Lee, Jin-Moo
    • The Journal of Korean Obstetrics and Gynecology
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    • v.32 no.4
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    • pp.1-13
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    • 2019
  • Objective: This study is aimed to investigate the anti-tumor metastasis by innate immunomodulating effects of crude polysaccharide isolated from Polygonati Rhizoma (CP-PR) on 4T1 breast cancer cells. Methods: CP-PR was isolated from Polygonati Rhizoma. Antimetastatic experiments were conducted in vivo mouse model by using 4T1 breast cancer cells. The cell viability of CP-PR was tested with normal spleen and 4T1 breast cancer cells. To observe the activation of macrophages with/without 4T1 breast cancer cells, production of tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), IL-10 and IL-12 were measured with enzyme-linked immunosorbent assay (ELISA), respectively. In addition, the lysis of YAC-1 cells and the production of granzymes were measured to observe the activation of natural killer (NK) cell. Results: Intravenous administration of CP-PR significantly inhibited metastasis of 4T1 breast cancer cells. In an in vitro cytotoxicity analysis, CP-PR affected the growth of normal spleen and 4T1 breast cancer cells above specific concentration. The production of $TNF-{\alpha}$, IL-6, IL-10 and IL-12 were significantly increased in macrophages with CP-PR. As compared with control, CP-PR showed significantly higher production of $TNF-{\alpha}$, IL-10 and IL-12 in macrophages co-cultured with 4T1 breast cancer cells. The lysis of YAC-1 cells and the production of granzymes were significantly up regulated by CP-PR. Conclusion: CP-PR appears to have considerable activity on the anti-metastasis by activation of innate immune system.

Effects of Cryptotympana pustulata on the expression of cytokine genes in human monocytes of THP-1 (선퇴가 인간의 THP-1 단핵구에서 사이토카인 유전자 발현에 미치는 영향)

  • An, Jong-Hyun;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.1
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    • pp.94-110
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    • 2010
  • Objective : This study was performed to evaluate the effect of immune reaction inductive substances such as phorbol-myristate-acetate(PMA), lipopolysaccharide(LPS), dermato-phagoides pteronyssus crude extract(DPE), dinitrochloro-benzene(DNCB) and Cryptotympana pustulata(CP), the Cryptotympana pustulata extracting substance at simultaneously on the translocation of nuclear factor-kappa B(NF-${\kappa}B$) towards to the nucleus and the mRNA expression patterns of various cytokine genes in Human acute monocytic leukemia cell line(THP-1 cells), monocytes of human. Experiment : To analyze cytokine genes expression patterns, the RT-PCR method was used, measuring tumor necrosis factor(TNF)-$\alpha$ that had been secreted during cell culture in the ELISA method. The morphological change in the cell observed during THP-1 cell culture was observed using a scanning electron microscope (SEM) and the quantitative distribution in the cell NF-${\kappa}B$ was analyzed through immunocytochemistry and a confocal microscopy. Result : CP showed different influences onto the mRNA expression patterns of cytokine genes with PMA, LPS. DPE and DNCB according to the types of immune inductive substances in the THP-1 cells. The expressions of inter-leukin(IL)-10, interferon(INF)-$\gamma$, TNF-$\alpha$ and monocyte chemoattractantant protein(MCP)-1 induced by PMA were suppressed by CP while the expression of transforming growth factor(TGF)-$\beta$ was promoted. Regarding the secretion pattern of TNF-$\alpha$ according to PMA processing, its secretion amount was increased by CP concurrent processing, in case of processing CP onto PMA and LPS, We discovered that the secretion amount of TNF-$\alpha$ was increased. Upon processing PMA and LPS on the THP-1 cell strain at the same time or either additionally processing CP thereon, the movement increase towards the nucleus from the NF-${\kappa}B$ cell cytoplasm, a transcription factor was able to be observed. Conclusion : In this study, Cryptotympana pustulata extracting substance was confirmed that it had an influence on expression patterns of cytokine genes according to the actions of a variety kinds of immune reaction inductive substances processed on the monocyte THP-1 cell of humans. Therefore, additional studies as for the immune adjusting function of Cryptotympana pustulata are considered to be able to offer important materials for curing immune abnormal diseases such as atopy dermatitis afterward.

Study of Anti-inflammatory Activity of 7 Herbal Prescription Effective for Sleep Disorders and Atopic Dermatitis (수면 장애 및 아토피피부염에 효과적인 7 가지 한약 처방의 항염증 활성 연구)

  • Choi, Min-Jin;Nguyen, Ly Thi Huong;Jeong, Jeonghwa;Park, Minyoung;Yoon, Joohee;Lee, Byung-Wook;Yang, In-Jun;Shin, Heung-Mook
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.35 no.2
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    • pp.56-63
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    • 2021
  • This study was conducted to find a candidate prescription with anti-inflammatory efficacy of 7 herbal prescriptions known to be effective in atopic dermatitis and sleep disorders in Korean medicine. The anti-inflammatory of the 7 herbal prescriptions extracts were evaluated by ELISA assay and Western blot assay. 7 herbal prescriptions, Seongyutang(SU), Danseontoetang(DST), Sotosajahwan(ST), Jisiljagyagsan(JJ), Seokchangpo(SCP), Wiryeongseon(WLS), Gogojohwan(GGJ) 30 % EtoH extract was used for in vitro experiments. In TNF-α + IFN-γ(TI) stimulated HaCaT cells, all 7 herbal prescriptions reduced TARC and MDC production, and JJ strongly reduced TARC and MDC production at 100 ㎍/ml concentration. SCP strongly reduced MDC production at 10 ㎍/ml and 100 ㎍/ml concentration. In addition, in substance P(SP)/CRH stimulated HMC-1 cells, JJ strongly inhibited VEGF production at both 10 and 100 ㎍/ml concentrations. In LPS/CRH stimulated Raw264.7 cells, all 7 herbal prescriptions significantly inhibited TNF-α. PMA + Ionomycin(PI)/CRH stimulated EL4 cells, SU significantly reduced IL-4 production at both concentrations of 10 and 100 ㎍/ml. WLS significantly reduced IL-17 production at both concentrations of 10 and 100㎍/ml. This suggests that 7 herbal prescriptions have anti-inflammatory effects by reducing inflammatory cytokines in keratinocytes, mast cells and macrophages caused by inflammation. Therefore, it is expected that 7 herbal prescriptions that are effective for sleep disorders and atopic dermatitis can be used as therapeutic materials for sleep disorders and diseases associated with atopic dermatitis.

Anti-invasion Effects of Calystegia soldanella Solvent Extracts and Partitioned Fractions on PMA-stimulated Fibrosarcoma Cells (갯메꽃 추출물과 유기용매 분획물의 암전이 억제 효과)

  • Son, Jaemin;Kim, Junse;Kim, Hojun;Seo, Youngwan
    • Journal of Life Science
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    • v.29 no.3
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    • pp.287-294
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    • 2019
  • Calystegia soldanella is distributed in coastal sand dunes and has high environmental adaptability; it is also known to be effective for anti-oxidant, anti-pyretic, anti-septic, and diuretic action. This study investigated the effect of crude extracts and organic solvent fractions of C. soldanella on MMP-2 and MMP-9 expression, MMP activity, and cell mobility in phorbol-12-myristate-13-acetate (PMA)-induced fibrosarcoma HT-1080 cells. C. soldanella was twice extracted, once with methylene chloride (MC) and once with methanol (MeOH). After the MC and MeOH extracts were combined, their suppressive effects on MMP-2 and MMP-9 expression, MMP enzymatic activity, and gene and protein expression were measured by gelatin zymography, enzyme-linked immunosorbent assay, reverse-transcription polymerase chain reaction, and western blot method. Cell mobility for the HT-1080 cells was observed by wound healing assay. The combined crude extracts showed a significant suppressive effects on MMP-2 and MMP-9 expression. To explore active inhibitory elements, the combined extracts were fractionated according to polarity into with n-hexane, 85% aqueous methanol, n-butanol, and water. Across these four solvent fractions, MMP-2 and MMP-9 activity and cell mobility in the HT-1080 cells were all strongly inhibited by the n-hexane fraction. These results suggest that C. soldanella extract and organic solvent fractions could be used as potent MMP inhibitors for effective anti-cancer treatments to suppress cancer invasion and metastasis.