• Title/Summary/Keyword: Freund's

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A Study on Antigenicity and Immunodepressive Activity of DA-125, A New Anthracycline Anticancer Agent (새로운 Anthracycline 항암제 DA-125의 항원성 및 면역독성에 대한 연구)

  • 백남기;강경구;김옥진;안병옥;이순복;김원배;양중익;정세영
    • Biomolecules & Therapeutics
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    • v.1 no.2
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    • pp.236-243
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    • 1993
  • Immunologic potential of DA-125, a new anthracycline antitumor antibiotic, was investigated using guinea pigs and mice. In antigenicity experiments, guinea pigs were sensitized subcutaneously with DA-125 or DA-125 incorporated in complete Freund's adjuvant (CFA) once a week for three weeks. No systemic anaphylaxis was induced by intravenous injection of DA-125 or DA-125 incubated with guinea pig serum after 3 weeks from the last sensitization. None of sera of these animals showed any passive cutaneous anaphylactic reaction (PCA) when DA-125 or DA-125 incubated with guinea pig serum was used as a challenging antigen in homologous PCA experiment. On the other hand the treatment of guinea pigs with ovalbumin Incorporated in CFA induced systemic anaphylactic reaction when challenged by intravenous injection of 5 mg/body of ovalbumin. Immunodiffusion test revealed no precipitating antibodies as detected in guinea pigs sensitized with DA-125. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with DA-125 or DA-125 mixed with aluminum hydroxide gel (Alum), None of sera showed positive reaction when DA-125 or DA-125 incubated with rat serum was used as a challenging antigen. Sera of animals immunized with a mixture of ovalbumin and alum showed positive PCA reaction when 5 mg/body of ovalbumin was injected as a challenging antigen. In lymphocyte proliferation tests, spleen lymphocyte proliferation to PHA and LPS was similarly impaired by 12 mg/kg of DXR or 36 mg/kg of DA-125, and the immunodepressive activity of DA-125 showed a dose-dependent manner. From these results, it could be concluded that immunosupression of DA-125 would be comparable to that of DXR and that DA-125 would not induce systemic allergic reaction in its clinical use.

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Development of Enzyme-Linked Immunosorbent Assay for Rapid and Sensitive Analysis of Biotin (Biotin의 분석을 위한 효소면역측정법(ELISA)의 개발)

  • 이경애;손동화;고영태
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.6
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    • pp.1152-1159
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    • 1998
  • In order to develop more rapid and reproducible analysis of biotin known as vitamin H, attempts were made to establish the condition for enzyme linked immunosorbent assay(ELISA) compared with traditional microbiological assay(MBA). Antibiotin and antiserum were obtained from the immunized rabbits injected with emulsion of biotin KLH conjugate and Freund's adjuvant. The antiserum showed cross reactivity on biocytin, a derivative of biotin, which is converted to biotin in intestine, at the rate of 177%(median inhibitory concentration(IC50)=12.58ppb), but not on other derivatives such as desthiobiotin, diaminobiotin and 2 imino biotin. Specific antibody for biotin was purified from the antiserum through protein A column and desalting column. The conditions of competitive direct ELISA (cdELISA) were established. Detection range of biotin concentration by cdELISA was 0.01∼300ng/ ml(ppb). In the spike test with milk, fruit flake and pine carrot juice, the correlation coefficient between two methods of MBA and ELISA was reliably consistent at the value of r=0.992. But detection of biotin by microbiological assay(MBA) was rather restricted in range and nonspecific. Detection range of biotin by MBA was 0.1∼0.5ng/ml(ppb). It showed cross reactivities on biocytin and desthiobiotin at the rate of 80.1% and 66.7%, respectively. In conclusion, ELISA revealed a significant improvement compared with MBA for the biotin detection in terms of sensitivity, detection range and cross reactivity. In addition, a variety of samples could be analyzed rapidly and conveniently at one time by using ELISA. These results strongly suggest that the ELISA is very promising for the practical application to detect biotin contents in a wide range of food stuffs.

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Anti-inflammatory Effect of Arbitrary Waveform Generator Treatment in Rats

  • Kim, Myung-Gyou;Lee, Se-Na;Seo, Il-Bok;Leem, Kang-Hyun;Ham, Kee-Sun;Kim, Hye-Kyung
    • Biomedical Science Letters
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    • v.14 no.1
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    • pp.39-45
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    • 2008
  • Inflammation is the complex biological response of injured tissues to harmful stimuli. A cascade of biochemical events propagates and matures the inflammatory response, involving the local vascular system, the immune system, and various cells within the injured tissue. The immune system is often involved with inflammatory disorders, demonstrated in both allergic reactions and some myopathies, with many immune system disorders resulting in abnormal inflammation. An Arbitrary Waveform Generator (AWG) is a piece of electronic test equipment used to generate electrical waveforms for the treatment of patients. The patients with gastritis and arthritis have been known to have a relatively favorable prognosis with AWG treatment. Accordingly, we examined the effects of AWG treatment in gastritis and arthritis animal model. The compound 48/80 was used to induce animal gastritis model. The tissue malone dialdehyde (MDA) and serum histamine levels, and the activity of superoxide dismutase (SOD) in stomach tissue were measured. The tissue MDA and serum histamine levels in AWG treated groups exhibited the decreased tendency compared with control group, whereas the tissue SOD activity was slightly increased. The Freund's complete adjuvant was used to induce animal arthritis model as well. The paw edema volume and the width of ankle joint were determined. The AWG treatment significantly decreased the paw edema volume after 5th day of treatment. Although further studies should be performed to confirm the effects of AWG treatment, present study suggest that AWG treatment might be used as a complementary treatment for the gastritis or arthritis treatment.

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Anti-inflammatory Effect of Isaria sinclairii Glycosaminoglycan in an Adjuvant-treated Arthritis Rat Model

  • Ahn, Mi Young;Jee, Sang Duck;Hwang, Jae Sam;Yun, Eun Young;Ahn, Kwang Seok;Kim, Yeong Shik
    • Toxicological Research
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    • v.29 no.3
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    • pp.195-201
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    • 2013
  • The anti-inflammatory effects of glycosaminoglycan (GAG) derived from Isaria sinclairii (IS) and of IS extracts were investigated in a complete Freund's adjuvant (CFA)-treated chronic arthritis rat model. Groups of rats were treated orally with 30 mg/kg one of the following: [1] saline control, extracts of [2] water-IS, [3] methanol-IS, [4] butanol-IS, [5] ethyl acetate-IS, or [6] Indomethacin(R) as the positive control for a period of two weeks. The anti-paw edema effects of the individual extracts were in the following order: water-IS ex. > methanol ex. > butanol ex. > ethyl acetate ex. The water/methanol extract from I. sinclairii remarkably inhibited UV-mediated upregulation of NF-${\kappa}B$ activity in transfected HaCaT cells. GAG as a water-soluble alcohol precipitated fraction also produced a noticeable anti-edema effect. This GAG also inhibited the pro-inflammatory cytokine levels of prostaglandin $E_2$-stimulated lipopolysaccharide in LAW 264.7 cells, cytokine TNF-${\alpha}$ production in splenocytes, and atherogenesis cytokine levels of vascular endothelial growth factor (VEGF) production in HUVEC cells in a dose-dependent manner. In the histological analysis, the LV dorsal root ganglion, including the articular cartilage, and linked to the paw-treated IS GAG, was repaired against CFA-induced cartilage destruction. Combined treatment with Indomethacin(R) (5 mg/kg) and IS GAG (10 mg/kg) also more effectively inhibited CFA-induced paw edema at 3 hr, 24 hr, and 48 hr to levels comparable to the anti-inflammatory drug, indomethacin. Thus, the IS GAG described here holds great promise as an anti-inflammatory drug in the future.

Active Systemic Anaphylaxis Test of Purified Bee Venom(Apis mellifera L.) (정제봉독의 아나필락시스 쇼크 반응 연구)

  • Han, Sang Mi;Hong, In Phyo;Woo, Soon Ok;Kim, Se Gun;Jang, Hye Ri;Park, Kyun Kyu;Chang, Young Chae
    • Korean Journal of Pharmacognosy
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    • v.46 no.3
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    • pp.203-207
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    • 2015
  • This study was performed to examine the antigenic potential of purified bee venom (Apis mellifera L., PBV) collected using bee venom collector. Antigenic potential of PBV was examined by active systemic anaphylaxis (ASA) in guinea pigs. PBV was subcutaneously administered at 0.025 and 0.05 mg/kg and also as a suspension with adjuvant (Freund's complete adjuvant, FCA). Ovalbumin (OVA) as a suspension with adjuvant was used to introduce positive control response. In the weight measurement and clinical observation, experimental groups didn't show any significant changes compared with control group. In the autopsy of body, the abnormalities of lung were detected only in the positive control. In the ASA test, experimental groups didn't show any symptoms of anaphylaxis like piloerection, hyperpnea and staggering gait. These results suggested that PBV didn't have antigenic potential in guinea pig.

Analgesic Effects of DA-5018, a New Capsaicin Derivative, after Subcutaneous Injection and Topical Application (새로운 캅사이신유도체 DA-5018의 피하주사 및 국소도포시 진통효과)

  • 김희기;배은주;신명수;손문호;김순희;김원배;양중익;공재양
    • Biomolecules & Therapeutics
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    • v.5 no.2
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    • pp.117-124
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    • 1997
  • The analgesic effects of DA-5018, a new caosaucin derivative, were evaluated in various experimental pain models. Drugs were administered subcutaneously or topically. When drugs were administered subcutaneously, 1) the $ED_{50}$ of DA-5018, morphine . HCI, capsaicin and acetaminophen were 0.091-2.0, 0.3-4.3, 1.4-26.5 and 45.4-643 mg/kg, respectively in various pain or inflammatory models including acetic acid writhing, formalin, tail flick, Randall-Selitto, hot plate and crouton oil-induced ear edema test, 2) the AD2 values (the dose for doubling of pain threshold of vehicle control) of DA-5018, capsaicin and ketoprpgin were 1.07 $\pm$ 0. 18, 23.47$\pm$4.46 and 2.97$\pm$0.43 mg/kg in Freund's complete adjuvant (FCA)-induced arthritic pain model. And by topical application, 1) neither DA-5018 0.3% cream nor Zostrix-HP (capsaicin 0.075%) were effective in formalin test, 2) although DA-5018 0.3% cream significantly inhibited the croton oil-induced ear edema being better than Zostrix-HP and Kenofen (ketoprofen 3%). 3) In FCA model, DA-5018 0.3% cream reversed the decreased pain threshold of arthritic rat from 136.4 g (day 0) to 289.0 g (day 5) and 250.1 g (day 10), which was similar to Zostrix-HP. These results suggest that DA-5018 administered subcutaneously has a potent and broad analgesic spectrum than nonsteroidal antiinflammatory drugs against acute and chronic pain, and by topical application it exerts comparable analgesic and antiinglammaatory effects to capsaicin cream.

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Thecooperative relationship between chemotherapy and the host immune response in immunosuppressed or immunostimulated mice infected with Fasciola hepatica (면역억압 또는 면역활성된 마우스에 간질(Fasciola hepatica)을 감염시킨 후 관찰되는 약물요법과 숙주의 면역기전과의 상호협력관계)

  • Shin, Sung-shik;Kim, Cheol-hee
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.575-585
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    • 2000
  • This study was performed to observe the influence of host immune response on the chemotherapy of mice experimentally infected with Fasciola hepatica. Following immunosuppression with prednisolone or immunoenhancement with Freund's complete adjuvant(FCA), mice were experimentally infected with 3 Fasciola hepatica metacercariae and treated with closantel at 1 week post infection. In the group of mice infected with metacercariae alone, 2 mice of 10 were dead at 10 weeks post infection(20% mortality), and adult flukes were recovered from the liver and the peritoneal cavity of the remaining 8 mice(100% infectivity). In the group of mice treated with prednisolone and infected with metacercariae, 8 of 10 mice died before euthanasia with a mean time of death earlier than the control group (p<0.05). In the group of immunosuppressed mice infected with metacercariae and treated with closantel 20mg/kg, 4 of 10 mice died before sacrifice. In the group of mice infected and treated with closantel 20mg/kg, mortality and infectivity was 10% and 30%, respectively. Similar results were observed in mice infected and treated with closantel 5mg/kg which resulted in 10% and 50% mortality and infectivity, respectively. These results indicated that the efficacy of closantel treatment was decreased in immunosuppressed mice, while the pathogenicity was increased. In immunoenhanced mice infected with metacercariae, on the other hand, the efficacy of chemotherapy with both 5mg/kg or 20mg/kg closantel resulted in only 10% infectivity. The results shown in this study strongly suggest that a close interaction between chemotherapy against F hepatica with closantel and the host immune system exists. Considering that fascioliasis is a zoonosis, treatment regimen against the infection to immunosuppressed patients may require a concurrent prescription of an appro-priate immuno-enhancing adjuvant.

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Antigenicity of DA-3030, a Recombinant Human Granulocyte-colony Stimulating Factor, in Guinea Pigs and Mice (Guinea pig 및 mouse에 있어서 인형 과립구 콜로니 자극인자 DA-3030의 항원성)

  • 백남기;강경구;이순복;김원배;양중익
    • Biomolecules & Therapeutics
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    • v.2 no.3
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    • pp.292-297
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    • 1994
  • This study was conducted to investigate antigenic potential of DA-3030, a recombinant human granulocyte-colony stimulating factor, in guinea pigs and mice. In the active systemic anaphylaxis test, the guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone did not show any anaphylactic reaction. In the homologous passive cutaneous anaphylaxis reaction, anti-DA-3030 antibody was not detected in guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone. On the other hand, the guinea pigs sensitized with 12.5 $\mu\textrm{g}$/heed of DA-3030 incorporated in Freund's complete adjuvant(FCA) or 1 mg/head of ovalbumin incorporated in FCA showed anaphylactic reaction. Anti-DA-3030 antibody was also detected in those guinea pigs. In immunodiffusion test using the sera sensitized with DA-3030 incorporated in FCA, precipitating antibodies were detected only in the sera sensitized with DA-3030 or DA-3030 incorporated in FCA showed. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone, none of the sera showed positive reaction. But sera of the animals immunized with 12.5 $\mu\textrm{g}$/head of DA-3030 incorporated in aluminum hydroxide gel(Alum) or 5 $\mu\textrm{g}$/head of ovalbumin incorporated in alum showed positive PCA reaction. DA-3030 did not cause anaphylactic shock or passive cutaneous anaphylaxis in guinea pigs and mice when given alone although DA-3030 incorporated in FCA or Alum induced anaphylactic shock and passive cutaneous anaphylaxis. From these results, it may be concluded the DA-3030 does not induce systemic allergic reaction when administered alone in its clinical use.

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The Analgesic Effect and Its Mechanism of Electroacupuncture in the Type II Collagen-induced Arthritis Rats : Mediation by Serotonergic Receptors (전침자극(電鍼刺戟)의 Collagen 유발 관절염(關節炎) 동물(動物)모델에 대한 진통효과(鎭痛效果) 및 기전(機轉)에 관한 연구(硏究) -Serotonergic Mechanism을 중심으로-)

  • Ryu, Seong-Ryong;Baek, Yong-Hyeon;Park, Dong-Suk
    • Journal of Acupuncture Research
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    • v.23 no.3
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    • pp.77-90
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    • 2006
  • Objectives : To evaluate the analgesic effect of electroacupuncture on Choksamni (ST36) in the collagen-induced arthritis rats and investigate the role played by serotonergic receptor subtypes $(5-HT_{1A},\;5-HT_{1B},\;5-HT_4)$ in the antinociceptive effect of electroacupuncture in the thermal hyperalgesia test. Methods : Immunization of male Sprague-Dawley rats with bovine type II collagen emulsified in incomplete Freund's adjuvant, followed by booster injection 14 days later induced collagen-induced arthritis (CIA). The thermal hyperalgesia was evaluated weekly with tail flick latency (TFL). In the fourth week after first immunization. EA stimulation (2Hz, 0.07mA, 0.3ms) was delivered into Choksamni for 20 minutes. We measured the analgesic effect of EA with TFL afer intraperitoneal injection of normal saline, WAYl00635, SB216641 and GR125487. Results : TFLs were gradually decreased in CIA as time elapsed after the immunization of arthrogenic collagen and the maximum value was reached from third to fifth week. EA stimulation on ST36 inhibited chronic inflammatory pain induced by CIA. The analgesic effect of EA was inhibited by pretreatment of $5-HT_{1A}$. antagonist (WAYl00635), $5-HT_{1B}$ antagonist (SB216641) and $5-HT_4$ antagonist (GR125487). Conclusion : Electroacupuncture has the analgesic effect on chronic inflammatory pain and its mechanism was mediated by $5-HT_{1A}$, $5-HT_{1B}$ and $5-HT_4$.

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Effects of Low Frequency Electro-Acupuncture (ST36) on the Adjuvant-Induced Arthritis in Rats (저빈도(低頻度) 족삼리(足三里) 전함랄극(電緘剌戟)이 흰쥐의 Adjuvant 유발(誘發) 관절염(關節炎)에 미치는 영향(影響))

  • Kim, Jong-Chul;Lee, Eun-Yong;Seo, Il-Beok;Koo, Sung-Tae;Choi, Sung-Mi;Kim, Ee-Hwa
    • Journal of Acupuncture Research
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    • v.23 no.6
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    • pp.207-219
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    • 2006
  • Objectives : This study was to investigate the effect of low frequency electro-acupuncture (EA) at ST36 acupoint on the Freund's Complete Adjuvant(FCA)-induced arthritis in rats. Methods : Arthritis was induced by intradermal injection of FCA into base of tail. Experimental groups were divided into 4 groups; Normal, Control, ST36 and Non-Acupoint(NA). ST36 group was treated by 2 Hz EA at ST36 acupoint. Body weight, paw edema volun1e and ankle joint thickness were measured after treatment. And we investigated the effects of 2 Hz EA via WBC count, segment neurophil, lymphocyte, PGE2 assay and NADPH-d histochemistry. Results and Conclusion : The mean of body weight and ankle joint thickness of ST36 group was increased compared with control group. The mean of paw edema volume, WBC count, segment neutrophil and lymphocyte of were not decreased with control group. The mean of PGE2 concentration and NADPH-d positive neurons were changed significantly compared with control group.

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