• Title/Summary/Keyword: cELISA

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The Effects of Alismatis Rhizoma Extract on Allergic Inflammation in RBL-2H3 Mast Cells and OVA/alum-Sensitized Mice (택사 (澤瀉, Alismatis Rhizoma) 추출물이 RBL-2H3 비만세포와 OVA/alum 감작 생쥐의 알레르기 염증 반응에 미치는 영향)

  • Song, Ji Hyun;Lee, Jin Hwa;Kim, Eun Jin;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.32 no.3
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    • pp.1-15
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    • 2018
  • Objectives Alismatis Rhizoma has been known to suppress inflammation and allergic reaction. However, the cellular target of Alismatis Rhizoma and its mechanism of action remain unclear. This study was designed to examine the effect of Alismatis Rhizoma extract (ALC) on the RBL-2H3 mast cells in vitro and on the OVA/alum sensitized mice ex vivo. Methods In the study, RBL-2H3 mast cells were cultured in minimal essential medium (MEM) for 24 hours, and treated separately with cyclosporin A and varying doses of ALC, and then stimulated with Phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) and Ionomycin ($0.5{\mu}M$). The levels of IL-13, IL-4 were measured by ELISA analysis. The mRNA levels of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$ were analyzed with Real-time PCR. Also, manifestations of MAPKs transcription factors and $NF-{\kappa}B$ p65 translocation were analyzed by western blotting in vitro. Subsequently, for ex vivo experiment, we induced allergic inflammation on Balb/c mice by OVA/alum and administered ALC orally. And we measured serum OVA-specific IgE level and IL-4, IL-13 in the splenocyte culture supernatant by ELISA analysis. Results ALC was shown to suppress mRNA expression of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$, and to inhibit the IL-13, IL-4 production. Also ALC reduced an activation of mast cells specific signal MAPKs transcription factors and $NF-{\kappa}B$ p65 from the western blot analysis in in vitro experiment. In ex vivo, ALC oral adminstration decreased the level of OVA-specific IgE in serum, and IL-4, IL-13 in the splenocyte culture supernatant. Conclusions ALC is shown to reduce inflammation and allergic response by suppressing Th2 cytokines through the regulation of transcription factors MAPKs and $NF-{\kappa}B$ p65 in mast cells. Administration of ALC suppressed OVA-specific IgE in ovalbumin allergy model through the inhibition of Th2 cytokine. In conclusion, ALC can be considered as an effective treatment for allergic diseases such as atopic dermatitis.

Changes in blastogenic responses and antibody titers of mice infected with Toxoplasma gondii (Toxoplasma gondii 감염 마우스에 있어서의 아세포화 반응 및 항체가 변화)

  • 신대환;이영하
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.125-132
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    • 1992
  • This study was performed to observe the cell-mediated and humoral immune responses in mice which were infected with Beverley, Fukaya and ME49 strain of Toxoplasma gondii, respectively. The blastogenic responses of splenocytes using $[^3H]-thymidine$ and serum antibody titers were measured weekly up to 10 weeks after infection. The blastogenic responses of splenocytes treated with concanavalin A and Tosoplasma Iysate were significantly declined in the 3 strain groups as compared with the non-infected group (P<0.05), however lipopolysaccharide-treated blastogenic responses were not significantly different between infected and non.infected groups. The serum IgG antibody titers in the three infected groups increased from 2 weeks after infection, and the serum IgM antibody titers increased until 4 weeks after infection. No significant differences were revealed in blastogenic responses and serum antibody titers among the 3 groups. The present study suggested that cell-mediated immune responses were involved in T. gondii infected mice and blastogenic responses of T Iymphocytes were inhibited in acute T. gondii infection.

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

  • Kim, Sun-Young;Kwon, Woo-Je;Kang, Sang-Mo
    • Journal of Applied Biological Chemistry
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    • v.53 no.4
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    • pp.239-247
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    • 2010
  • Helicobacter pylori (H. pylori) is the main causal bacteria occurring stomach diseases such as gastritis and gastric ulcer. These bacteria are found in most adults' stomach. Especially, 60~90% of H. pylori is found in Korean stomach. As a lot of curing methods have been applied to remove H. pylori and certain effects have been done but it's impossible to remove it perfectly with the existing medicines for curing. Therefore, it's very urgent to develop a certain substance showing more excellent effect than the existing medicines. In this study, it wasfound that organic acids can be accessed easily, inserted into mouth for curing and has excellent sterilizing effect among the substances showing excellent antibiotic power. Among them, acetic acid showed the most excellent effect. To confirm the refraining power against H. pylori, we performed tests through in vitro contact testing methods by concentration and tsta. In the result, H. pylori were terminated perfectly in the solution of acetic acid more than 0.3% within 1 minute. With a base of the result of In vitro test, It was performed in vivo test. As the results, H. pyloriwere terminated perfectly on 0.2% solution of acetic acid from the result of confirming through urease test, ELISA method and RT-PCR. Therefore, the result of this research will be very useful information in developing the functional foodstuffs using acetic acid in order to terminate H. pylori on the people's stomach, who suffers from H. pylori.

The Effect of Polycan-Calcium Gluconate Complex on Inflammatory Mediators from Periodontitis Patients (Polycan-Calcium Gluconate 복합제에 의한 염증인자의 감소 효과)

  • Kim, Ji-Hye;Kim, Ki-Rim;Jin, Hye-Jung;Im, Sang-Uk;Song, Keun-Bae;Choi, Youn-Hee
    • Journal of dental hygiene science
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    • v.14 no.2
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    • pp.223-229
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    • 2014
  • The aim of this study was to examine the effect of polycan-calcium gluconate complex on the levels of immune and inflammatory mediators in gingival crevicular fluid and serum from patients with periodontitis. A total of 39 patients with periodontitis took placebo (placebo group) or polycan-calcium gluconate complex (treatment group) twice a day for 4 weeks. At baseline and 4 weeks, gingival crevicular fluid (GCF) and blood was collected from each subjects. The secretion level of interleukin $(IL)-1{\beta}$ in GCF was measured by enzyme-linked immunosorbent assay (ELISA). Western blot analysis was conducted to determine the expression of matrix metalloproteinase (MMP)-8 and tumor necrosis factor $(TNF)-{\alpha}$ in GCF. Serum samples were analysed for MMP-8 by ELISA and C-reactive protein (CRP) by turbidimetric immuno assay. MMP-8 and $TNF-{\alpha}$ were significantly decreased in the treatment group at 4 weeks. The level of $IL-1{\beta}$ in the treatment group was significantly lower than those of the placebo group. No differences were observed in CRP levels. Taken together, these data indicate that taking of polycan-calcium gluconate complex led to reduction of inflammatory biomarkers in serum and GCF of periodontitis patients.

Reduction of the Antigenicity of Powdered Milk by Gamma Irradiation (감마선 조사에 의한 분유 단백질의 항원성 저감화)

  • Shin, Ji-Hye;Jeong, Seok-Geun;Han, Gi-Sung;Jang, Ae-Ra;Chae, Hyun-Seok;Yoo, Young-Mo;Ahn, Chong-Nam;Lee, Ju-Woon;Jo, Cheo-Run;Lee, Wan-Kyu;Ham, Jun-Sang
    • Food Science of Animal Resources
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    • v.28 no.3
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    • pp.306-311
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    • 2008
  • The aim of this work was to examine the reduction in antigenicity of milk proteins in powdered milk by gamma irradiation which is increasingly used for food safety. Skim milk powder samples were exposed to irradiation doses of 1, 5, and 10 kGy. A greater reduction of ${\alpha}_{S1}$-casein and ${\beta}_{A1}$-casein was found than ${\alpha}_{S0}$-casein and ${\beta}_{A2}$-casein by capillary electrophoresis. Competitive indirect ELISA and passive cutaneous anaphylaxis tests using guinea pigs showed a reduction in antigenicity of powdered milk by 10kGy gamma irradiation. These results indicated that gamma irradiation reduce allergenicity of milk proteins by structural changes of ${\alpha}_{S1}$-casein and ${\beta}_{A1}$-casein, and can be useful for dairy products.

Effect of Eff Component on Stability of IgY Antibody Acitivity to In Vitro Digestion (In Vitro 소화시 lgy항체 활성의 안정성에 대한 계란 성분의 효과)

  • 이승배;최석호
    • Food Science of Animal Resources
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    • v.20 no.4
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    • pp.303-310
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    • 2000
  • In vitro 소화시 계란 성분이 anti-Y ruckeri IgY 항세 활성을 안정화시키는데 어떡해 관여하는지를 SDS-PAGE와 ELISA로 조사한 결과는 다음과 같다. Anti-Y ruckeris IgY 항체와 난황 및 난백으로 혼합 후 펩신으로소화시킨 후 경우 1시간 후 난황 및 난백이 혼합된 시료에서 모두 anti-Y ruckeri IgY 항체의 heavy chain과 light chain이 분해는 되었지만 어느 정도 밴드를 관찰할 수있었으며, anti-Y, ruckeri IgY 항체 활성 측정한 결과 난황시료는 35%, 난백시료는 61%의 항체 활성을 유지시켰다. Anti-Y ruckeri IgY 항체와 오보알부민, 오보뮤우신, 라이소자임 및 오보뮤코이드를 혼합 한 후 펩신으로 소화시킨 경우 1시간 우에는 anti-Y, ruckeri IgY항체의 heavy chain의 밴드르 fdjsm 정도 볼 수 있는 형태로 나타났으며, anti-Y, ruckeri IgY 항체의 light chain는 오보뮤우신 및 오보뮤코이드가 홉합되어 펩신으로 소화시킨 경우 1시간 후에는 밴드를 거의 볼수 없었으나 오보알부민 오보트란스훼린 및 라이소자임에 혼합된 시료에서 밴드를 관찰할수 있었다. 특히 라이소자임과 오보트란스훼린의 경우 펩신 호화 2시간 우에도 anti-Y, ruckeri IgY 항체의 light 밴드가 관찰되었다. 펩신소화 1시간 후에 난백성분이오보알부민, 오보뮤우신, 라이소자임, 오보트란스웨린 및 오부뮤코이드 중에서 단지 오보트란스훼린만이 38%의 anti-Y, ruckeri IgY 항체 활서을 보인후 2시간 후에도 15% 정동의 활성을 나타내었다. anti-Y, ruckeri IgY항체와 난백 및 전란을 혼합한 다음 무지개송어 위 추출액으로 소화시 2시간 후에 난백은 14%, 전란은 69%로 anti-Y, ruckeri IgY 항체 활성을 유지시킨 것으로 나타났다.X> $e_{I}$ WPi_BE_QE]]]]]로 상징하며 WLWQ에 적용되는 몇 가지 제약을 관찰하고 이를 일반적인 언어원리로 설명한다. 첫째, XP는 주어로만 해석되는데 그 이유는 XP가 목적어 혹은 부가어 등 다른 기능을 할 경우 생략 부위가 생략의 복원 가능선 원리 (the deletion-up-to recoverability principle)를 위배하기 때문이다. 둘째, WLWQ가 내용 의문문으로만 해석되는데 그 이유는 양의 공리(the maxim of quantity: Grice 1975) 때문이다. 평서문으로 해석될 경우 WP에 들어갈 부분이 XP의 자질의 부분집합에 불과하므로 명제가 아무런 정보제공을 하지 못한다. 반면 의문문 자체는 정보제공을 추구하지 않으므로 앞에서 언급한 양의 공리로부터 자유롭다. 셋째, WLWQ의 XP는 주제어 표지 ‘는/-은’을 취하나 주어표지 ‘가/-이’는 취하지 못한다(XP-는/-은 vs. XP-가/-이). 이는 IP내부 에 비공범주의 존재 여부에 따라 C의 음운형태(PF)가 시성이 정해진다는 가설로 설명하고자 했다. WLWQ에 대한 우리의 논의가 옳다면, 본 논문은 다음과 같은 이론적 함의를 기닌다. 첫째, WLWQ의 존재는 생략에 대한 두 이론 즉 LF 복사 이론과 PF 삭제 이론 중 전자의 입장을 지지한다. 둘째, WP를 XP로부터 복원할 때 부분 자질만 복사된다. 이는 어휘가 통사층위로 들어온 이후에도 어휘 자질들이 완전히 동결되는 것이 아니라 계속 지시될 수 있다는 가설을 지지한다.

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Transgenic Siberian Ginseng Cultured Cells That Produce High Levels of Human Lactoferrin (인체 락토페린 생산 형질전환 가시오갈피 배양세포)

  • Jo Seung-Hyun;Kwon Suk-Yoon;Kim Jae-Whune;Lee Ki-Teak;Kwak Sang-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.209-215
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    • 2005
  • Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

Study on the Changes in Allergen and Allergenicity Originated from Shrimp by Physical Treatments (물리적 처리에 의한 새우유래 Allergen 및 Allergenicity 변화)

  • Kim, Seong-Mi;Park, Jin-Gyu;Kim, Koth-Bong-Woo-Ri;Lee, Ju-Woon;Byun, Myung-Woo;Park, Sun-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.8
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    • pp.990-996
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    • 2006
  • This research was conducted to evaluate the changes in allergenicity of shrimp by physical treatments using competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). Shrimp was subjected to physical treatments such as high hydrostatic pressure (HHP), sonication, autoclave and microwave. Heat-stable protein (HSP) purified from raw shrimp was used as a major allergen. The binding ability of monoclonal IgG and shrimp-allergic patients' IgE to HSP treated with HHP decreased, increasing the pressure up to 400 MPa. Especially, it became less than 50% at 400 MPa. The binding ability of mAb to HSP treated with sonication (10, 20, 30 and 60 min) decreased with the treated time. Especially, it became less than 60% with the treatment for 60 min. The allergenicity change of HSP treated with autoclave and microwave little decreased. The binding ability of mAb to HSP during the treatment for 20 min became more than 70%. The results suggest that allergenicity of HSP in raw shrimp be more easily lost by HHP or sonication treatment than by autoclave or microwave treatment.

The Effects of Somatid on the Cytotoxicity of Cancer Cells and Human Papillomavirus Type 16 E6 and E7 Oncogenes (생기액(生肌液)의 세포독성 및 자궁경부암 바이러스 (HPV 16 type) 암 유발인자 E6와 E7의 작용에 미치는 효과)

  • Joung, Ok;Cho, Young-Sik;Cho, Cheong-Weon;Lee, Kyung-Ae;Shim, Jung-Hyun;Cho, Min-Chul;Lee, Hong-Soo;Yeom, Young-Il;Kim, Sang-Bom;Park, Sue-Nie;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.44 no.4
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    • pp.340-346
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    • 2000
  • 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 types 16 and 18) and cervical cancer has been well known. An extract of natural products, named as Somatid, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Somatid 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 Somatid inhibited the in vitro biding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Somatid 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 Somatid. Our data suggested that Somatid 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 caused by HPV.

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Serum IgE levels in rats infected with Parugonimus westermani (폐흡충(Paragonimus westermani) 감염백서에서의 혈청내 IgE 항체가의 변동)

  • Sin, Myeong-Heon;Ryu, Jae-Suk;Min, Deuk-Yeong
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.397-402
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    • 1991
  • Paragonimus westermani is a common fluke in Korea. The present study aimed to determine serum total IgE and specific IgG levels in experimental paragonimiasis of rats. Each Wistar rat was inoculated orally with 20~25 metacercariae of P. westermani from Cambaroides similis. Before and after infection(1,2,3,4,6,8 weeks) of P. westermani, the blood was collected from the retro- orbital venous plexus of rats and kept serum at $-70^{\circ}C$. Serum total IgE and specific IgG levels were determined by the capture and conventional enzyme-linked immunosorbent assay, respectively. The results were as follows; 1. Serum IgE values were increased to 0. 18${\pm}$0.042 at 2 weeks, $0.28{\pm}0.151$ at 4 weeks and 0.43${\pm}0.055$ at 8 weeks after infection. The absorbances of non-infected rats ranged $0.07{\pm}0.021~0.12{\pm}0.025$. 2. Specific IgG values were slightly increased at 3 weeks ($0.20{\pm}$0.032) and gradually increased up to 8 weeks($0.31{\pm}0.067$) after infection. The absorbances of non-infected rats ranged $0.11{\pm}0.035~0.18{\pm}0.019$. The present results suggested that p. westermani could elevate serum IgE and specific IgG antibodies in Wistar rats which were not a good definitive host.

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