• Title/Summary/Keyword: Chemical antigen

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The Radio-Immunoassay Method for Ginsenoside $Rg_1$ of Korean Ginseng

  • Han, Byung-Hoon;Han, Yong-Nam;Sankawa, Ushio;Akyama, Minko;Kawashima, Koichiro
    • Korean Journal of Pharmacognosy
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    • v.11 no.3_4 s.43
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    • pp.133-140
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    • 1980
  • In order to develop the radio-immunoassay procedure for the ginsenoside $Rg_1$ we prepared the $Rg_1-BSA$ conjugate and $Rg_1-tyramine$ conjugate by condensing the $Rg_1-azide$, which was prepared by a series of six step chemical modification of the $Rg_1-side$ chain, with bovine serum albumin(BSA) or with tyramine. Rabbits were immunized by repeated injection of $Rg_1-BSA$ conjugate with Freund's Complete Adjuvant for 5 month long to obtain very potent $anti-Rg_1$ serum. The radio-labelled haptene was prepared by direct radio-iodination $(125_J)$ of $Rg_1-tyramine$ according to the chloramine-T method. The radio-immunoassay procedure was successfully furnished by using DCC method (dextran coated charcoal) and the anti-body titer of the anti-serum was found as being $1600{\sim}3200$ by using 15000cpm tracer per test. Calibration test using non-labelled $Rg_1$ showed linear competetive binding response in the $(8-300){\times}34pg$. range of non-labelled $Rg_1$. The cross reaction test using 19 ginsenoside analogues enabled us a full structure-activity analysis on the antigen-antibody reaction that the anti-body in the serum would recognize the full structure of ginsenoside $Rg_1$ except the side chain moiety.

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Cellular Mechanism of Newly Synthesized Indoledione Derivative-induced Immunological Death of Tumor Cell

  • Oh, Su-Jin;Ryu, Chung-Kyu;Baek, So-Young;Lee, Hyun-Ah
    • IMMUNE NETWORK
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    • v.11 no.6
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    • pp.383-389
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    • 2011
  • Background: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell. Methods: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC. Results: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and $100{\mu}M$) with carleticulin induction. And EY-6 induced the secretion of IFN-${\gamma}$ but not of TNF-${\alpha}$ from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation). Conclusion: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

Role of Glutathione Conjugation in 1-Bromobutane-induced Immunotoxicity in Mice

  • Lee, Sang-Kyu;Lee, Dong-Ju;Jeon, Tae-Won;Ko, Gyu-Sub;Yoo, Se-Hyun;Ha, Hyun-Woo;Kang, Mi-Jeong;Kang, Won-Ku;Kim, Sang-Kyum;Jeong, Tae-Cheon
    • Toxicological Research
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    • v.26 no.2
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    • pp.101-108
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    • 2010
  • Halogenated organic compounds, such as 1-bromobutane (1-BB), have been used as cleaning agents, agents for chemical syntheses or extraction solvents in workplace. In the present study, immunotoxic effects of 1-BB and its conjugation with glutathione (GSH) were investigated in female BALB/c mice. Animals were treated orally with 1-BB at 375, 750 and 1500 mg/kg in corn oil once for dose response or treated orally with 1-BB at 1500 mg/kg for 6, 12, 24 and 48 hr for time course. S-Butyl GSH was identified in spleen by liquid chromatography-electrospray ionization tandem mass spectrometry. Splenic GSH levels were significantly reduced by single treatment with 1-BB. S-Butyl GSH conjugates were detected in spleen from 6 hr after treatment. Oral 1-BB significantly suppressed the antibody response to a T-dependent antigen and the production of splenic intracellular interlukin-2 in response to Con A. Our present results suggest that 1-BB could cause immunotoxicity as well as reduction of splenic GSH content, due to the formation of GSH conjugates in mice. The present results would be useful to understand molecular toxic mechanism of low molecular weight haloalkanes and to develop biological markers for exposure to haloalkanes.

Chronic and Low Dose Exposure to Nonlyphenol or Di(2-Ethylhexyl) Phthalate Alters Cell Proliferation and the Localization of Steroid Hormone Receptors in Uterine Endometria in Mice

  • Kim, Juhye;Cha, Sunyeong;Lee, Min Young;Hwang, Yeon Jeong;Yang, Eunhyeok;Choi, Donchan;Lee, Sung-Ho;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.23 no.3
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    • pp.263-275
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    • 2019
  • Based on our preliminary results, we examined the possible role of low-dose and chronic-exposing of the chemicals those are known as endocrine disrupting chemical (EDC), on the proliferation of uterine endometrium and the localization of steroid receptors. Immunohistochemical or immunofluorochemical methodology were employed to evaluate the localization of antigen identified by monoclonal antibody Ki 67 protein (MKI67), estrogen receptor 1 (ESR1), estrogen receptor 2 (ESR2), and progesterone receptor (PGR). In $133{\mu}g/L$ and $1,330{\mu}g/L$ di(2-ethylhexyl) phthalate (DEHP) and $50{\mu}g/L$ nonylphenol (NP) groups, the ratio of MKI67 positive stromal cells was significantly increased but not in $500{\mu}g/L$ NP group. The ratios of MKI67 positive glandular and luminal epithelial cells were also changed by the chronic administration of NP and DEHP in tissue with dose specific manner. ESR1 signals were localized in nucleus in glandular and luminal epithelia of control group but its localization was mainly in cytoplasm in DEHP and NP administered groups. On the other hand, it was decreased at nucleus of stromal cells in $1,330{\mu}g/L$ DEHP group. The colocalization patterns of these nuclear receptors were also modified by the administration of these chemicals. Such a tissue specific and dose specific localization of ESR2 and PGR were detected as ESR1 in all the uterine endometrial tissues. These results show that the chronic lows-dose exposing of NP or DEHP modify the localization and colocalization of ESRs and PGR, and of the proliferation patterns of the endometrial tissues.

Antiproliferative properties of luteolin against chemically induced colon cancer in mice fed on a high-fat diet and colorectal cancer cells grown in adipocyte-derived medium

  • Park, Jeongeun;Kim, Eunjung
    • Journal of Nutrition and Health
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    • v.55 no.1
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    • pp.47-58
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    • 2022
  • Purpose: Obesity and a high-fat diet (HFD) are risk factors for colorectal cancer. We have previously shown that luteolin (LUT) supplementation in HFD-fed mice markedly inhibits tumor development in chemically induced colon carcinogenesis. In this study, we evaluated the anticancer effect of LUT in the inhibition of cell proliferation in HFD-fed obese mice and HT-29 human colorectal adenocarcinoma cells grown in an adipocyte-derived medium. Methods: C57BL/6 mice were fed a normal diet (ND, 11.69% fat out of total calories consumed, n = 10), HFD (40% fat out of total calories consumed, n = 10), HFD with 0.0025% LUT (n = 10), and HFD with 0.005% LUT (n = 10) and were subjected to azoxymethane-dextran sulfate sodium chemical colon carcinogenesis. All mice were fed the experimental diet for 11 weeks. 3T3-L1 preadipocytes and HT-29 cells were treated with various doses of LUT in an adipocyte-conditioned medium (Ad-CM). Results: The weekly body weight changes in the LUT groups were similar to those in the HFD group; however, the survival rates of the LUT group were higher than those of the HFD group. Impaired crypt integrity of the colonic mucosa in the HFD group was observed to be restored in the LUT group. The colonic expression of proliferating cell nuclear antigen and insulin-like growth factor 1 (IGF-1) receptors were suppressed by the LUT supplementation in the HFD-fed mice. The LUT treatment (10, 20, and 40 µM) inhibited the proliferation and migration of HT-29 cells cultured in Ad-CM in a dose-dependent manner, as well as the differentiation of 3T3-L1 preadipocytes. Conclusion: These results suggest that the anticancer effect of LUT is probably due to the inhibition of IGF-1 signaling and adipogenesis-related cell proliferation in colon cancer cells.

Studies on the proliferative activity and the expression of c-myc and ras of preneoplastic hepatocytes in rats (랫드 전암 간세포의 증식성과 c-myc, ras의 발현에 관한 연구)

  • Cho, Ho-seong;Park, Nam-yong
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.117-129
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    • 2000
  • This study was carried out to examine proliferative activity and expression of c-myc oncoprotein and p2lras in normal and preneoplastic rat livers induced by an in vivo mid-term chemical carcinogenesis assay. Sixty, six-week-old male specific pathogen free Sprague-Dawley male rats were randomly divided into five groups. Group I was received a single intraperitoneal(IP) dose(200mg/kg) of diethylnitrosamine(DEN). Group 2(10 rats) was operated partial hepatectomy(PH) and Group 3 was received IP(200mg/kg) DEN, fed two weeks later with 500ppm of phenobarbital(PB). Group 4 was received IP(200mg/kg) DEN, fed two weeks later 500ppm(PB) and PH at week 3 after the onset of experiment. While group 5(20 rats) was not treated and used as a control group. All the rats were sacrificed at age 14 weeks except 10 rats from group 5 were sacrificed at the onset of experiment. Livers of all rats were examined for 5-bromo-2'-deoxyuridine(BrdU) incoporation, proliferating cell nuclear antigen(PCNA), silver-binding nucleolar organizer regions(AgNORs) counts per nucleus and expression of c-myc oncoprotein and p21ras. Both the number and area of the preneoplastic lesions were significantly(p<0.01) compared to other groups. A significant(p<0.01) increase in immunoreactive cells were detected in preneoplastic hepatocytes in Groups 3 and 4 by PCNA and BrdU immunohistochemical stain. The number of the positive cells were significantly(p<0.05) lower in normal 14-week-old rats than those of 6-week-old rats. The results showed that proliferative activity of the hepatocytes was increased by treatment with DEN, PH and PB. Meanwhile, AgNORs counts per nucleus were significantly(p<0.05) increased in the preneoplastic hepatocytes of rats in both groups 3 and 4. The expression of c-myc oncoprotein and p21ras were more readily localized within the hepatic preneoplastic lesions such as hyperplastic nodules. Especially, group 4 showed significantly (p<0.05) overexpressed levels compared to groups 1 and 3. These findings suggest that PCNA, BrdU and AgNORs are significantly increased and c-myc oncoprotein and p21ras are significantly overexpressed in hepatic preneoplastic lesions induced by mid-term carcinogenesis. So these parameters can be an effective markers for hepatic prencoplastic lesions.

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Synergic Anti-Pruritic and Anti-Inflammatory Effects of Scutellariae Radix plus Flos Loncerae Extracts in Rat Peritoneal Mast Cell and Chemical Antigen-Induced Mice (렛트 복강 비만세포와 화학항원 유도 알레르기 마우스에서 황금과 금은화 추출물의 항가려움 및 항염증 상승효과)

  • Mok, Ji Ye;Jeon, In Hwa;Kim, Hyeon Soo;Shin, Jun Ho;Park, Yong Gyoun;Jang, Seon Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.1
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    • pp.83-91
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    • 2013
  • Pruritus is a unpleasant symptom in the skin that provokes the act of or desire to scratch. Mast cells are important effector cells in allergic reactions such as pruritus and inflammation. The purpose of this study was undertaken to investigate the synergic anti-pruritic and anti-inflammatory effects of Scutellariae Radix (SB) plus Flos Loncerae (FL) extracts in rat peritoneal mast cells (RPMCs), pruritogen-induced scratching mice and 2,4-dinitrofluorobenzene (DNFB)-induced allergic mice. We investigated the effect of SB, FL and SB plus FL extracts on the production of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$, and histamine in RPMCs, on the scratching behavior in ICR mice, and skin clinical serverity and inflammatory mediators in DNFB-induced allergic hairless mice. RPMCs stimulated with PMA plus A23187 or compound 48/80 significantly increased TNF-${\alpha}$, IL-$1{\beta}$ or histamine production compared with media control. However, TNF-${\alpha}$ IL-$1{\beta}$ or histamine levels increased by PMA plus A23187 or compound 48/80 treatment were significantly inhibited by SB, FL in a dose-dependent manner. Especially, SB plus FL pretreatment had a synergic inhibitory effects on stimulator-induced cytokines (TNF-${\alpha}$ and IL-$1{\beta}$) and histamine production. Moreover, SB plus FL administration had a synergic inhibitory effects on the scratching behavior induced by pruritogen (compound 48/80, histamine, serotonin, substance P) in ICR mice. Furthermore, SB plus FL administration had a synergic inhibitory effects on skin damage, inflammatory mediators, leukocyte and mast cell infiltration induced by DNFB in hairless mice. These results suggest that SB plus FL administration has a potential use as a medicinal plant for treatment against itching and inflammation-related skin disease.

Effects of Gamma Irradiation on Queso Blanco Cheese (퀘소블랑코 치즈의 감마선 조사 처리 효과)

  • Jeong, Seok-Geun;Noh, Young-Bae;Shin, Ji-Hye;Han, Gi-Sung;Chae, Hyun-Seok;Yoo, Young-Mo;Ahn, Jong-Nam;Lee, Ju-Woon;Jo, Cheor-Un;Lee, Wan-Kyu;Ham, Jun-Sang
    • Journal of Dairy Science and Biotechnology
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    • v.25 no.1
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    • pp.15-20
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    • 2007
  • Effects of gamma irradiation on chemical, microbiological, and immunological changes of Queso Blanco cheese were investigated. Although Queso Blanco cheese was made by heat pasteurization at 85$^{\circ}$C and addition of acid without lactic starter culture, total bacterial counts and lactic acid bacterial counts of control cheese were 7.65${\pm}$0.04 and 7.64${\pm}$0.02 log CFU/mL, respectively. It was thought that this microbial growth was due to the incomplete inactivation of raw milk by the heat treatment, resulting into growth during the pressing and the drying process. It demonstrated the possibility that if heat- and acid-resistant hazard microbes are present in raw milk, they can grow during the processes. Lactic acid bacterial counts of the irradiated cheese were 5.45${\pm}$0.02 log CFU/mL at 1kGy, 2.12${\pm}$0.12 log CFU/mL at 2kGy, and not detected at 3kGy or higher doses. The reduction of antigenicity by gamma irradiation was not found. It might be caused by the fact that most whey proteins of milk, a major antigen in milk, were already denaturated by heat process and removed during the draining.

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Studies on Analysis Method of T-2 Toxin by ELISA (ELISA에 의한 T-2 toxin의 분석법에 관한 연구)

  • 오유진;장성재;윤여표
    • Journal of Food Hygiene and Safety
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    • v.3 no.2
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    • pp.65-73
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    • 1988
  • T-2 toxin is one of mycotoxins produced by fungi such as Fusarium spp. and possesses a potent cytotoxicity to eukaryotic cell. The contamination of mycotoxins in cereals and feedstuffs is one of the great concerns in health authorities. Therefore, the development of the specific, sensitive and simplified analysis method for T -2 toxin is required. During more than ten years, several chemical and biological analysis methods were proposed and applied for the detection and quantification of T-2 toxin. TLC, GLC-FID and GC-MS are widely employed, but these methods required numerous clean-up procedures before analysis, and the detection limit for T-2 toxin is more than 10 ppb. Biological analysis methods with dermal tissues and cultured cells are not specific to T-2 toxin, since T-2 toxin and other related derivatives possess a similar toxicological activity although their relative activity is different each otber. Based on tbe specific reaction between antibody and antigen, the authors tried to introduce the immunochemical methods for determination of T-2 toxin. The enzyme-linked immunosorbent assay method using monoclonal antibody for T-2 toxin was applied to analyse T-2 toxin. The detection limit of T-2 toxin by ELISA method was 0.1 ppb. The correlation between ELISA and GC-MS method on these samples was very high. ELISA method developed for the detection and quantification of T -2 toxin in this paper possesses simplicity, high sensitivity and specific for T-2 toxin. Furthermore, the ELISA method with T-2 toxin monoclonal antibody was an excellent tool for the screening of Fusarium spp. which was suspected to produce T-2 toxin.

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