• 제목/요약/키워드: ELISA method

검색결과 596건 처리시간 0.03초

Daily Amperometric Monitoring of Immunoglobulin E in a Mouse Whole Blood: Model of Ovalbumin Induced Asthma

  • Lee, Ju Kyung;Yoon, Sung-hoon;Kim, Sang Hee
    • 전기화학회지
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    • 제25권1호
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    • pp.13-21
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    • 2022
  • There is an increasing interest in monitoring of specific biomarker for determining progression of a disease or efficacy of a treatment. Conventional method for quantification of specific biomarkers as enzyme linked immunosorbent assay (ELISA) has high material costs, long incubation periods, requires large volume of samples and involves special instruments, which necessitates clinical samples to be sent to a lab. This paper reports on the development of an electrochemical biosensor to measure total immunoglobulin E (IgE), a marker of asthma disease that varies with age, gender, and disease in concentrations from 0.3-1000 ng/mL with consuming 20 µL volume of whole blood sample. The sensor provides rapid, accurate, easy, point-of-care measurement of IgE, also, sequential monitoring of total IgE with ovalbumin (OVA) induced mice is another application of sensor. Taken together, these results provide an alternative way for detection of biomarkers in whole blood with low volumes and long-term ex-vivo assessments for understanding the progression of a disease.

Enhancing the Effect of Aronia Extract on Hyaluronic Acid Synthesis through Liposome Formation

  • Youn, Young Han
    • 인간식물환경학회지
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    • 제23권4호
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    • pp.465-473
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    • 2020
  • Background and objective: Aronia melanocarpa, called black chokeberry, is a natural product belonging to the family rosaceae, and is known to contain polyphenolic antioxidants including cyanidin-3-galactoside, cyanidin-3-arabinoside, cyanidin-3-xyloside, and cyanidin-3-glucoside Because of the abundance of anthocyanins, Aronia has been studied to be used in various industries. Methods: Aronia melanocarpa extract was treated 24 hours a day to RAW 264.7 cells with inflammations induced by LPS. After extracting total RNA, the amount of inflammatory cytokine expression was measured using RT-PCR. After processing the Aronia liposome using Aronia extract and the layer-by-layer electrostatic deposition method in keratinocyte cells at the same time, we checked the synthesis of Hyaluronic acid enhanced through the formation of Aronia liposome using ELISA. Results: The treatment of Aronia extract in inflammation-induced RAW 264.7 cells conducted to check the anti-inflammatory efficacy of Aronia extract inhibited inflammatory cytokines including TLR4, TNF-α, IL-1β, COX-2, and iNOS and increased the mRNA expression of HAS2 genes related to moisturizing. Based on the anti-inflammatory and moisturizing effect of Aronia extract, the Aronia liposome technology was introduced to Aronia extract to produce Aronia liposome. Conclusion: The liposome formation of Aronia extract is expected to be used as a functional material in treating various inflammatory skin diseases by controlling the moisture content of the corneocytes by increasing the expression rate of genes associated with the synthesis of hyaluronic acid, while retaining the efficacy of its components.

Adenine, new anti-wrinkle agent.

  • Kim, Y. J.;Kim, Y. S.;S.Y. Eom;Kim, J. H.
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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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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쑥부쟁이 추출물의 아세틸콜린에스테라제 저해 및 항산화 활성 (Acetylcholinesterase Inhibitory and Antioxidant Properties of Aster yomena Extract)

  • 배종섭;김태훈
    • 대한본초학회지
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    • 제24권4호
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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.

Lipopolysaccharide로 활성화된 마우스 대식세포에서 애엽(艾葉) 물추출물의 면역활성 연구 (Effect of Water Extract from Artemisiae Argi Folium on Mouse Macrophage Stimulated by LPS)

  • 박완수
    • 대한본초학회지
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    • 제24권1호
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    • pp.151-157
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    • 2009
  • Objectives : The purpose of this study is to investigate the effect of Water Extract from Artemisiae Argi Folium (WAAF) on mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods : Cell viabilities were measured by MTT assay. And the intracellular productions of hydrogen peroxide (H2O2) were measured by dihydrorhodamine 123 assay. TNF-$\alpha$ and IL-6 production from Raw 264.7 were measured by ELISA method. Results : The results of the experiment are as follows. 1. WAAF significantly increased the cell viability compared to the control group (treated with LPS only) at the concentrations of 10, 50, 100, 200, 400 ug/mL. 2. WAAF significantly increased the intracellular production of H2O2 compared to the control group at the concentrations of 50, 100, 200 ug/mL. 3. WAAF significantly decreased the production of TNF-$\alpha$ compared to the control group at the concentrations of 100, 200 ug/mL. 4. WAAF significantly decreased the production of IL-6 compared to the control group at the concentrations of 50, 100, 200 ug/mL. Conclusions : WAAF could be supposed to have the immune-modulating activity related with the macrophage's immunoactivity.

Detection of Listeria Species by Conventional Culture-Dependent and Alternative Rapid Detection Methods in Retail Ready-to-Eat Foods in Turkey

  • Emine Dincer
    • Journal of Microbiology and Biotechnology
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    • 제34권2호
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    • pp.349-357
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    • 2024
  • Foodborne pathogens, like Listeria monocytogenes, continue to inflict substantial financial losses on the food industry. Various methods for detecting Listeria in food have been developed and numerous studies have been conducted to compare the different methods. But, in recent years, new Listeria species have been identified, and currently the genus comprises 26 species. Therefore, it would be a more accurate approach to re-evaluate existing detection methods by considering new species. The present investigation involved the analysis of 42 ready-to-eat (RTE) foods, encompassing a variety of food categories, such as mezes, salads, dairy products, and meat products, with the aim of ascertaining the presence of Listeria. Among the traditional culture-dependent reference methods, the ISO 11290 method was preferred. The process of strain identification was conducted with the API Identification System. Furthermore, to ascertain the existence of L. monocytogenes and Listeria spp., the samples underwent additional analysis employing the VIDAS Immunoassay System, ELISA, and RT-PCR methodologies. Thus, four alternative approaches were employed in this study to compare not only the different methods used to determine Listeria while taking into account the newly identified Listeria species, but also to assess the compliance of retail RTE food items with microbiological criteria pertaining to the genus Listeria. Based on the conducted analyses, L. monocytogenes was conclusively determined to be present in one sample. The presence of Listeria spp. was detected in 30.9% of the samples, specifically in Turkish cig kofte, sliced salami, and salads.

Interferon-$\gamma$가 사람 폐포대식세포의 결핵균 탐식과 활성화에 미치는 영향 (The Effect of IFN-$\gamma$ on the Phagocytosis of Mycobacterium tuberculosis and Activation of Human Pulmonary Alveolar Macrophage)

  • 박재석;김재열;이귀래;유철규;김영환;한성구;심영수
    • Tuberculosis and Respiratory Diseases
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    • 제45권1호
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    • pp.36-44
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    • 1998
  • 연구배경: IFN-$\gamma$는 단핵식세포를 활성화시키며 여러 종류의 세포내 세균에 대한 숙주의 방어기전에 중요한 역할을 하는 것으로 알려져있다. 그러나 사람에 있어서 IFN-$\gamma$의 항 결핵 효과와 작용기전에 대해서는 거의 알려진 바가 없다. 본 연구에서는 결핵의 발병기전에서 IFN-$\gamma$의 역할을 알아보기 위해 폐포대식세포의 결핵균 탐식과 TNF-$\alpha$ 생산에 IFN-$\gamma$가 미치는 영향을 알아보았다. 방 법: 활동성 폐질환이 없는 8명의 사람에게서 얻은 기관지 폐포세척액에서 폐포대식세포를 표면흡착법으로 분리하여 결핵균과 같이 배양하면서 ($1{\times}10^6$ cells/ml, $3{\times}10^7$ bacteria/ml) 배양액에 IFN-$\gamma$(300U/ml), LPS(0.5ug/ml), 자가혈청(10%)을 첨가하여 2시간 배양 후 항산성 염색(modified Kynion method)을 하여 결핵균을 탐식한 폐포대식세포를 관찰하였다. 그리고 폐포대식세포배양액에 IFN-$\gamma$(300U/ml), MTB($1{\times}106bacteria/ml$) and LPS(0.5ug/ml)를 각각 첨가하여 24시간 배양 후 상층액에서 TNF-$\alpha$의 농도를 ELISA method로 측정하였다. 그리고 IFN-$\gamma$(300U/ml), LPS(0.5ug/ml)로 24시간 자극한 폐포대식세포의 결핵균 탐식율도 관찰하였다. 결 과: IFN-$\gamma$는 폐포대식세포의 결핵균 탐식율을 증가시키지 않았으며(percentage of PAM-phagocytosed MTB: control: $22.1{\pm}4.9$, IFN-$\gamma$: $20.3{\pm}5.3$), 폐포대식세포를 24시간 자극하였을 때 폐포대식세포의 TNF-$\alpha$의 생산을 증가시키지 않았다 (control: $21{\pm}38pg/ml$, IFN-$\gamma$: $87{\pm}106pg/ml$). 그리고 IFN-$\gamma$로 24 시간 전처치한 폐포대식세포의 결핵균 탐식율 또한 증가하지 않았다(control: $24.5{\pm}9.5$, IFN-$\gamma$: $23.4{\pm}10.1$). 결 론: IFN-$\gamma$는 폐포대식세포의 결핵균 탐식과 TNF-$\alpha$ 생산에 영향을 미치지 않는다.

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치근단 및 치수병변 환자에서의 TNF-α와 β, IL-1β 및 TGF-β1의 수준과 근관내 특정 black pigmented bacteria와의 연관성에 관한 연구 (LEVELS OF TNF-α,-β, IL-1β, TGF-β1 AND THEIR RELATIONSHIP WITH THE PRESENCE OF SPECIFIC BLACK PIGMENTED BACTERIA IN PERIAPICAL AND PULPAL DISEASES)

  • 변호영;임성삼;박동성
    • Restorative Dentistry and Endodontics
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    • 제24권1호
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    • pp.1-12
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    • 1999
  • Bacterial infection of the pulp results in the development of a periapical lesion with the concomitant resorption of periapical bone. The cytokines are believed to play an important role in this matter. The purpose of this study was to find the relationship among the presence of black pigmented bacteria, the levels of cytokines(TNF-${\alpha}$, -${\beta}$, IL-$1{\beta}$, and TGF-${\beta}1$), and the amount of bone resorption in periapical and pulpal diseases. For the purpose, the patients were grouped into chronic apical pathosis, acute apical pathosis, acute pulpitis, and a healthy control group. Root canal samples were taken from periapical tissue exudates during routine endodontic treatment, and the venous blood was taken from each patients. The samples were processed to measure local and systemic levels of the cytokines using enzyme linked immunosorbent assay(ELISA). Bacterial content of Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella nigrescens were measured by indirect immunofluorescence method and the size of the periapical lesions were measured from the radiographs. The following results were obtained: 1. The levels of bone resorptive cytokines(TNF-${\alpha}$, TNF-${\beta}$, and IL-$1{\beta}$) in exudates from acute and chronic apical pathoses were significantly higher than those from acute pulpitis and the normal pulps(p<0.05). 2. IL-$1{\beta}$ were the highest among the bone resorptive cytokines in apical pathoses. However, no statistical difference between acute and chronic lesions were found(p>0.05). 3. The levels of TGF-${\beta}1$ in exudates from acute pulpitis and chronic apical pathoses were significantly higher than those from acute apical pathoses and the normal pulps(p<0.05). However, there were no significant correlations among the levels of bone resorptive cytokines. 4. The levels of TNF-${\beta}$ in serum were significantly higher than those from the exudates while serum TGF-${\beta}1$ concentrations were significantly lower(p<0.05). 5. Exudates from the canals in which the P. nigrescens were detected showed significantly higher levels of IL-$1{\beta}$ than those from the canals without the microorganism(p<0.05). 6. There were no significant correlations among the levels of the cytokines, the amount of bone destruction, and the presence of acute and chronic symptoms(p>0.05).

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초산에 의한 마우스 위의 Helicobacter pylori 살균효과 (The Antibiotic Effect of Acetic acid on Helicobacter pylori)

  • 김선영;권우제;강상모
    • Journal of Applied Biological Chemistry
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    • 제53권4호
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    • pp.239-247
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    • 2010
  • Helicobacter pylori(H. pylori)는 위 감염증의 주된 원인균으로, 위염 및 위궤양을 일이키는 균으로 알려져 있다. 대부분의 성인 남녀의 위에서 발견이 되면, 특히 한국 성인의 위에서 60~90% 정도 발견된다. H. pylori을 제균하기 위해 많은 치료 방법들이 시도되었고 어느 정도의 제균 활성은 있었으나, 완벽한 제균은 기존의 치료 약물로는 어렵다. 따라서 기존의 약물보다 더 효과가 좋은 물질의 개발이 시급한 상황이다. 본 연구에서는 항균 능력이 뛰어난 물질 중, 쉽게 접할 수 있고 경구투여로 치료가 용이하여 약재나 식품으로 응용할 수 있는 물질들중 유기산들이 살균능력이 뛰어난 것을 알았다. 그 중 초산이 가장 뛰어나 살균효과를 보였다. 초산의 H. pylori에 대한 억제 능력을 확인하기 위해 농도별, 시간별로 in vitro 접촉실험법으로 실험을 시행하였다. 그 결과 0.3% 이상의 초산 용액에서는 H. pylori가 1분 이내에 100% 사멸되었다. In vitro 실험 결과를 토대로 BALB/c mouse를 이용한 in vivo 실험을 하였다. H. pylori를 경구투여로 위해 접종하였다. 다음 감염된 것을 확인하고 초산용액을 각 군에 해당하는 농도에 준하여 0.1mL를 1일 1회 혹은 3회 경구투여하였다. 초산에 의한 H. pylori의 사멸 정도를 재배양 실험법, urease test 및 ELISA법으로 확인한 결과 초산용액 0.2% 식이에서 모두 사멸되었다. RT-PCR법으로 검정한 결과는 초산용액 0.2%에서 밴드가 나타났지만 이는 사멸된 균의 것으로 보이며, 0.3%에서 존재하지 않는 것으로 나타났다. RT-PCR법에서 0.2% 초산용액 식이에서는 H. pylori가 확인되는 것은 H. pylori의 사균이 위벽에 붙어 있다가 RT-PCR법으로 확인되는 것으로 보였다. 따라서 이 연구 결과가 세상 사람들의 절반이상이 감염되어 있는 H. pylori를 퇴치하는 데 매우 유용한 자료가 될 것이다. 또한 초산을 이용한 기능성 식품 개발에도 중요한 자료로써 활용될 것으로 생각된다.

목단피의 NF-κB와 MAPKs 억제를 통한 Pro-inflammatory Cytokines 활성에 미치는 효과 (The Effect of Moutan Cortex on Pro-inflammatory Cytokines through NF-κB & MAPKs pathway in HMC-l)

  • 박주호;김경준
    • 한방안이비인후피부과학회지
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    • 제22권2호
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    • pp.1-18
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    • 2009
  • Objective : Moutan Cortex (the root bark of Paeonia suffruticosa Andr.) is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how MC(Moutan Cortex) affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted MC on the mast cell-mediated inflammatory responses. Method : We observed the effect of MC on compound 48/80-induced histamine release of rat peritoneal mast cells and the effect of administering MC on PCA in rat. We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of MC. The TNF-$\alpha$ protein levels were analysised by Western blot. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-$\kappa$B, phospho-I$\kappa$B and MAPKs were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by luciferase assay. Result : 1. Enzyme immunoassay indicated that MC suppressed histamine secretion of rat peritoneal mast cells. 2. In PCA dependent on IgE, MC had anti-allergic effect of the internal surface of rat skin. 3. Western blot indicated that MC decreased TNF-$\alpha$ protein levels. 4. ELISA indicated that MC decreased TNF-$\alpha$, IL-6 but MC had no significant effect on IL-8 in HMC-1 cells. 5. RT-PCR indicated that MC decreased TNF-$\alpha$, IL-8 but MC had no significant effect on IL-6 in HMC-l cells. 6. Western blot indicated that MC suppressed the induction of MAPKs, NF-$\kappa$B & phospho-I$\kappa$B activity in HMC-1 cells. 7. Luciferase assay indicated that MC suppressed the PMA plus A23187-induced NF-$\kappa$B promoting activityin HMC-1 cells. Conclusion : In this study, we have found that MC is an inhibitor of NF-$\kappa$B, MAPKs & cytokines on the mast cell-mediated inflammatory responses.

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