• Title/Summary/Keyword: TNBS-method

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Comparison of Determination Methods of Amino Nitrogen in Salt-Fermented Anchovy Sauce

  • Cho Young-Je;Kim Tae-Jin;Choi Yeung-Joon
    • Fisheries and Aquatic Sciences
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    • 제4권3호
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    • pp.144-149
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    • 2001
  • In other to establish the exact determination method of amino nitrogen (AN) in salt-fermented fish sauces, we determined the AN in fish sauces according to the measuring methods and also investigated the main factors influencing on determination method of AN. AN in salt-fermented anchovy sauce increased linearly as fermentation progressed, and was shown the highest amount measuring by the Formol method, followed by the trinitrobenzene sulfonic acid (TNBS) method and the Copper-salt method. AN concentration in anchovy sauces fermented for 12 months was $88.2\%$ and $77.6\%$ for the TNBS method and the Copper-salt method, respectively, on the basis of Formol method. The ratio of AN/total nitrogen (TN) in anchovy sauce fermented for 12 months was higher than that in commercial anchovy sauces. The determination of AN in anchovy sauce by the TNBS method was not affected by salt concentration, and slightly affected by heating. The effect of MSG on AN contents by Copper-salt method was shown higher than those by the Formol method and the TNBS method. The TNBS method was adaptable to measure the content of AN in fish sauce by this study.

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천추(天樞) 상응부위에 구진약침(灸津藥針) 자극(刺戟)이 TNBS로 유도(誘導)된 크론병에 미치는 영향 (Effects of Moxi-tar Herbal Acupuncture at Cheonchu (ST25) on Crohn's Disease Induced by TNBS in Mices)

  • 김영태;안성훈;김재효;손인철
    • Korean Journal of Acupuncture
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    • 제25권2호
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    • pp.159-177
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    • 2008
  • Objectives : Crohn's disease is a severe chronic inflammation that is treated mainly by immunosuppression, which often has serious side effects. There is need to develop new therapeutic methods or drugs that have few side effects in order to treat this disease. Acupuncture with Moxi-tar at Cheonchu (ST25) has anti-inflammatory properties, but the mechanism of its anti-inflammatory actions is unclear. We investigated the protective effects and speculated the mechanisms of acupuncture with Moxi-tar at ST25 on trinitrobenzene sulfonic acid (TNBS) induced colitis in mice which is a well known Crohn's disease animal model. Methods : 5 % TNBS was treated at day 1 and day 7 into rectum of mice. To investigate therapeutic effects of acupuncture with Moxi-tar at ST25, acupuncture was carried out on day 3, and day 6. For the data analysis, we observed macroscopic and microscopic findings of the colon. Weight and width of the colon, degree of damage, changes of body weight, and myeloperoxygenase (MPO) activity were checked. For analysing protein expression, we carried out immunohistochemical staining and Western blot. For analysing mRNA expression, RT-PCR was carried out. Results : TNBS induced damages on the colon of mice, while acupuncture of Moxi-tar at ST25 suppressed TNBS mediated damages similar to those on the colons of mice in the control (not treated with TNBS) group. The average body weight of TNBS treated mice (77.4%) was decreased compared with that of the control mice (105%), and acupuncture with Moxi-tar at ST25 suppressed the loss of body weight caused by TNBS (from 77.4% to 95.3%). TNBS induced infiltration of immune cells in all layers of the colon while acupuncture with Moxi-tar at ST25 suppressed infiltration of immune cells caused by TNBS. Furthermore, acupunctured with Moxi-tar at ST25 suppressed macro-, micro- colonic damages caused by TNBS. Acupunctured with Moxi-tar at ST25 dramatically improved the clinical and histopathological symptoms such as the increase in weight of the distal colon and the MPO activity in TNBS-induced colitis. Acupuncture with Moxi-tar at ST25 down-regulated the nuclear transcription factor kappa B ($NF-{\kappa}B$) activity and suppressed tumor necrosis factor-a (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-1${\beta}$), and intracellular adhesion molecule-1 (ICAM-1) expressions caused by TNBS. Conclusions : Acupuncture with Moxi-tar at ST25 helps recovery from the TNBS-induced colonic damage by down-regulation of $NF-{\kappa}B$ activity and suppressing of TNF-${\alpha}$, IL-1${\beta}$, and ICAM-1 expressions. This may be an important method for the treatment of Crohn's disease.

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까나리액젓 중의 아미노태질소 측정 (Estimation of Amino-nitrogen Content in Salt-fermented Sand Lance Sauce)

  • 조형제;송민우;임영선;최영준
    • 수산해양교육연구
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    • 제12권2호
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    • pp.213-223
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    • 2000
  • 액젓중의 아미태질소 함량의 정확한 측정방법을 확립하기 위하여, 아미노태질소 함량 측정방법으로 많이 사용되고 있는 포르몰법, TNBS법 그리고 동염법(銅鹽法)으로 까나리액젓의 숙성기간 및 시판액젓중의 아미노태질소 함량을 측정 비교하고, 액젓중의 아미노태질소 함량 측정시에 식염농도, 가열 및 MSG(monosodium glutamate) 첨가의 영향에 대하여 검토하였다. 까나리액젓의 숙성기간중의 아미노태질소 함량은 숙성기간에 따라서 일정하게 증가하였으며, 포르몰법이 가장 높았고 TNBS법 및 동염법(銅鹽法)의 순서였다. 12개월 숙성후의 아미노태질소 함량은 포르몰법을 기준으로 TNBS법은 88.7% 그리고 동염법(銅鹽法)은 75.2%였으며. 시판 까나리액젓중의 아미노태질소 함량도 포르몰법이 가장 높았고 TNBS법 및 동염법(銅鹽法)의 순서였다. 탈염 액젓중의 아미노태질소 함량은 포르몰법 및 TNBS법은 비슷한 값이었고, 동염법(銅鹽法)은 약간 낮았으며, 식염 첨가량이 많을 수록 포르몰법 및 동염법(銅鹽法)은 감소하였으나 TNBS법은 거의 일정하였다. 가열에 의한 영향은 거의 없었으며 포르몰법이 가장 높았고. TNBS법 및 동염법(銅鹽法)의 순서였다. MSG 첨가에 따른 아미노태질소 함량은 첨가농도에 비례하여 일정하게 증가하였으며, 숙성기간 및 가열중의 결과와 유사하게 포르몰법이 가장 많았고, TNBS법 및 동염법(銅鹽法)의 순서였다.

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TNBS 유발 대장염(大腸炎)에 의한 체중 및 혈액상 변화에 대한 인삼(人蔘) 약침(藥鍼)의 효과 (Effects of Ginseng Radix herbal acupuncture on TNBS-induced colitis in rats)

  • 김종성;천상우;박상연;박성익;김경식;김재효;구성태;손인철
    • Korean Journal of Acupuncture
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    • 제23권4호
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    • pp.147-156
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    • 2006
  • Objectives : The purpose of the present study is to evaluate the effects of herbal acupuncture with Ginseng Radix for the treatment to intestinal disease in the rat with 2, 4, 6-trinitrobenzenesulfonic acid (TNBS) induced colitis. Methods : All animals were subjected to the injection of saline $(300{\mu}{\ell},\;500{\mu}{\ell})$ for a study control or TNBS $(300{\mu}{\ell},\;500{\mu}{\ell})$ into the lumen of the colon, 8cm proximal to the anus through the intestine. Ginseng Radix herbal acupuncture ($20mg/m{\ell},\;0.4m{\ell}$) were injected to the both $Hapgok(LI_4)$ acupoints at 2nd injection of TNBS in rats. Thus, the body weight, RBC count, WBC count, total protein, IgG levels and IgM levels were observed to study the effects of Ginseng Radix herbal acupuncture. Results : Ginseng Radix herbal acupuncture on $Hapgok(LI_4)$ for TNBS-induced colitis inhibited the body weight loss rate but did not affect RBC and WBC counts. Furthermore, it inhibited the reduction of total protein concentration and serum IgG and IgM levels in TNBS induced colitis were recovered. Conclusions : Herbal acupuncture with Ginseng Radix helps recover the TNBS-induced colonic damage and may be an important method for treatment of the colitis.

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TNBS로 유도된 대장염에서 곡지의 구진약침 효과 (Effects of Moxi-tar Herbal Acupuncture of LI11 on inflammatory bowel disease induced by TNBS in mice)

  • 송문영;박상연;김재효;안성훈;김경식;손인철
    • Korean Journal of Acupuncture
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    • 제25권3호
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    • pp.147-166
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    • 2008
  • Objectives : Crohn's disease (CD) is characterized by a chronic relapsing inflammation of the bowel in which proinflammatory cytokines play an important perpetuating role. Methods : Mice (preventive animal model of gliotoxin) were treated with 5 % 2,4,6-trinitrobenzenesulfonic acid (TNBS) at day 1 and day 7. To investigate preventive effects of acupuncture with Gujin at $LI_{11}$, acupuncture was carried out at day -1, day 1, day 3. And, to investigate therapeutic effects, acupuncture with Gujin was carried out at day 3, day 5, day 7. For the data analysis, we checked weight and width of colon, diarrhea, edema, survival rate, changes of body weight, and myeloperoxygenase (MPO) activity. For analysing protein expression, we carried out immunohistochemical staining and Western blot and we analyzed mRNA expression by RT-PCR. Results : Colon of TNBS treated mice was erosive and shortening compared with the colon of control mice and induced damages of colon epithelial cell layer and induced infiltration of immune cells in all layer of colon. Acupuncture of gujin at $LI_{11}$ in preventive mode suppressed macorscopic damages such as erosive and shortening of colon by TNBS and damages of intestinal epithelial cells and infiltration of immune cells in the colon. The average weight of 5 cm distal colon was increased in TNBS treated mice (758${\mu}g$) compared with in control mice (112${\mu}g$) and width of distal colon was also increased in TNBS treated mice (4.9mm) compared with in control mice (1.3mm). Acupuncture with Gujin at $LI_{11}$ in preventive and therapeutic mode suppressed increase of colon weight and width by TNBS. TNBS induced edema of colon and diarrhea and Acupunctured with Gujin at $LI_{11}$ in preventive and therapeutic mode ameliorated these symptom by TNBS. In preventive and therapeutic mode, the effects of acupuncture with Gujin at $LI_{11}$ were increasing the motility, suppressing body weight decreasing, suppressing MPO activity, reducing expressing of TNF-${\alpha}$, IL-1b, and ICAM-1 in colon compared with that by TNBS Conclusions : This study demonstrates that acupuncture with Gujin at $LI_{11}$ represents a potential therapeutic method of Crohn's disease.

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생쥐의 급 ${\cdot}$ 만성 대장염에 대한 곡지의 구진 약침 효과 (Effects of Moxi-tar Herbal Acupuncture of LI11 on inflammatory bowel disease induced by TNBS in mice)

  • 송문영;권오상;장재훈;장재영;박성익;김재효;김경식;손인철
    • Korean Journal of Acupuncture
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    • 제24권4호
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    • pp.131-149
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    • 2007
  • Objectives : The purpose of the present study is to evaluate the effects of herbal acupuncture (HA) with Moxi-tar for the treatment to intestinal disease in mice with 2, 4, 6 - trinitrobenzenesulfonic acid (TNBS) induced colitis. Methods : Mice were administered with 5% TNBS at day 1 and day 7. To investigate effects of HA with Moxi-tar at LI11, treatments were carried out at day -1, day 1, day 3, day 5, and day 7. It was checked on the weight and width of colon, diarrhea, edema, survival rate, changes of body weight, and myeloperoxygenase (MPO) activity. Furthermore, we carried out immunohistochemical staining and Western blot and analyzed mRNA expression by RT-PCR. Results : HA of Moxi-tar at LI11 in preventive mode suppressed macroscopic damages and damages of intestinal epithelial cells and infiltration of immune cells in the colon by TNBS. HA in early and preventive mode ameliorated various symptoms by TNBS. TNBS injection increased MPO activity in colon while HA in preventive mode suppressed increase of MPO activity. HA down-regulated NF-kB activity and reduced expression of TNF-a, IL-1b, and ICAM-1 in colon of TNBS treated mice. Similar to experiment at colon, HA down-regulated NF-kB activity and reduced expression of TNF-a, IL-1b, and ICAM-1 by TNBS in mesenteric lymph node. HA in therapeutic mode suppressed errosion and shortening of colon and MPO activity by TNBS and suppressed mRNA expression of TNF-a, IL-1b, and ICAM-1 in the colon. Conclusions : This study demonstrates that HA with Moxi-tar at LI11 represents a potential therapeutic method of inflammatory bowel diseases.

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Osmometry에 의한 Hemoglobin 가수분해도의 신속한 측정 (Rapid Determination of Degree of Hydrolysis for Hemoglobin by Osmometry)

  • 채희정;인만진;김동호;강인규;오남순
    • Applied Biological Chemistry
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    • 제44권3호
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    • pp.143-147
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    • 2001
  • Hemoglobin(Hb) 가수분해도(degree of hydrolysis, DH)를 간편하고 신속하게 측정하고자 보편적으로 이용되는 TNBS법(trinitrobenzene sulfonic acid method)에 의한 가수분해도와 osmometer를 이용한 방법의 상관관계를 효소반응 조건에 따라 조사하였다. 다양한 ph 범위$(pH\;7.5{\sim}10.0)$에서 DH에 대한 두 방법의 상관계수($R^2$값)는 $0.974{\sim}0.991$로 매우 높은 값을 보였으며, 각각 측정한 DH 사이의 비율($DH_{OSM}/DH_{TNBS}$$1.438{\sim}1.656$의 범위에 있었다. Hb을 가수분해하여 heme-iron을 얻기 위한 특정 반응조건인 pH 7.5, $50^{\circ}C$에서 Esperase와 Fla-vourzyme을 각각 사용한 경우와 Esperase와 Flavourzyme을 혼용한 경우에서도 두 방법간의 DH 비율은 효소의 종류 및 처리방법에 무관하게 1.658을 나타내어 Esperase만을 사용한 경우와 매우 유사하였고 두 방법간의 상관계수는 0.982이었다. 따라서, 두 방법간의 상관관계식으로부터 osmometer를 이용한 Hb의 가수분해도를 간편하고 신속하게 측정할 수 있었다.

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TNBS로 유도된 만성 염증성 대장염에 대한 대장유(大腸兪) 구진약침(灸津藥鍼)의 효과 (Moxi-tar Herbal Acupuncture of BL25 Acupoint Ameliorates TNBS-Induced Colitis in Mice)

  • 백대봉;권오상;최원종;김재효;전희영;김경식;손인철
    • Korean Journal of Acupuncture
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    • 제24권3호
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    • pp.149-164
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    • 2007
  • Objectives : Inflammatory bowel disease (IBD) is a chronic relapsing inflammatory disease in the gastrointestinal tract. The sources and pathologic mechanisms of IBD are still unknown. Moreover conventional therapies for IBD are not always effective, and they often have serious side effects. The purpose of this study is to examine the effect of Moxi-tar herbal acupuncture in IBD affected mice. Methods : Mice were treated with 5 % 2, 4, 6 - trinitrobenzenesulfonic acid (TNBS) on day 1 and day 7. To assume the preemptive effect and therapeutic effect, herbal acupuncture was practiced with Moxi-tar at BL25 (Daejangsu) on day 0, day 3, and day 6. The end of day in treatment with Moxi-tar herbal acupuncture, the mortality and the inflammatory factors of the colon were measured by the various methods. Results TNBS induced high mortality but herbal acupuncture with Moxi-tar at BL25 sup-pressed the mortality caused by TNBS. TNBS induced infiltration of immune cells in all layers of the colon and increased myeloperoxygenase (MPO) activity, while the treatment with Moxi-tar herbal acupuncture at BL25 suppressed the infiltration of immune cells and the increase of MPO activity caused by TNBS to normal levels, Herbal acupuncture with Moxi-tar regulated $NF-{\kappa}B$ activity, which is an important factor for the pathogenesis of chronic colitis, and reduced the expressions of $TNF-{\alpha}$, $IL-1{\beta}$, and ICAM-1 in the colons of TNBS treated mice. Furthermore herbal acupuncture suppressed macro- and micro- colonic damages caused by TNBS. Conclusions : This study demonstrates that herbal acupuncture with Moxi-tar at BL25 isa potential preemptive and/or therapeutic method targeting the chronic IBD.

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합곡에 대한 애엽 약침이 흰쥐 대장염 완화에 미치는 효과 (Artemisiae argyi herbal acupuncture of Hapgok (LI4) ameliorates TNBS-induced colitis in rats)

  • 변기원;임용수;구성태;김재효;김경식;손인철
    • Korean Journal of Acupuncture
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    • 제24권1호
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    • pp.79-94
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    • 2007
  • Objectives : Ulcerative colitis and Crohn's disease are chronic inflammatory disease of the gastrointestinal tract was identified mainly for diarrhea, stomachache, hemafecia. The source and pathologic mechanism about ulcerative colitis and crohn's disease are still unknown but stress, infection, a genetic factor, environmental factors etc. may be the cause of chronic inflammatory disease of the gastrointestinal tract recently. The purpose of the present study is to evaluate the effects of herbal acupuncture with Artemisiae Argyi for the treatment to intestinal disease in the rat with 2, 4, 6 - trinitrobenzenesulfonic acid (TNBS) induced colitis. Methods : Artemisiae Argyi Herbal acupunctures were infected in HapGok (LI4) which is used to treat for intestinal disease in clinic. All animals were subjected to the injection of saline (300 ${\mu}l$, 500 ${\mu}l$) for a study control and TNBS (300 ${\mu}l$, 500 ${\mu}l$) into the lumen of the colon, 8cm proximal to the anus through the intestine. Artemisiae Agryi' Herbal (20 mg/ml, 0.4 ml) acupunctures were injected to the LI4 (both hands) at the secondary injection time of TNBS in rats. And body weight, RBC count, WBC count, total protein, Paw edema rate, rate of protein leakage into CMC-pouch fluid, IgG levels and IgM levels were observed to study the effects of Artemisiae Argyi' Herbal acupuncture in HapGok (Ll4). Results : Artemisiae Argyi' Herbal acupuncture in HapGok (Ll4) on TNBS-induced colitis inhibit the body weight lose rate but not RBC and WBC count. Furthermore, it inhibited the reduction of total protein concentration, paw edema, rate of protein leakage into CMC-pouch fluid, IgG levels and IgM levels. Conclusions : Herbal acupuncture with Artemisiae Argyi helps recover the TNBS-induced colonic damage and may be an important method for treatment of the colitis.

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발효단백질의 유리아미노산 정량 (Conditions of Quantitative Analysis for free Amino Acid in Fermented Proteins)

  • 류홍수;문정혜;이강호
    • 한국식품영양과학회지
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    • 제17권2호
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    • pp.136-143
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    • 1988
  • 발효단백질 중의 유리아미노산을 효과적으로 정량하기 위하여 정량방법(추출조건, 제단백(除蛋白) 및 발색법)에 따른 영향을 검토하였다. 낙농발효품의 수용성 유리아미노산은 $75^{\circ}C$, 40분, 대두발효품은 $40^{\circ}C$, 3시간에서 효과적으로 추출되었다. 여러 침전방법(95% ethanol 처리, $100^{\circ}C$, 3분 가열 및 1% picric acid phthaldialdehyde로 발색시켜 D.L.-lysine을 표준아미노산으로 삼아 정량, 계산하였을 때가 아미노산자동분석기에 의한 결과에 가장 접근하였으나 ethanol로 침전시킨 뒤 OPDA로 발색시킨 결과도 이와 유사하였다. TNBS(2,4,6-trinitrobenzensulfonic acid)에 의한 발색정량에는 ethanol 침전처리가 가장 효과적이었으나 종래의 동감법(銅監法)(copper salt method, Spies and Chamber, 1958.)에 의한 결과는 다른 비색정량 및 아미노산자동분석기에 의한 결과에 비하여 시료에 따른 과소의 차이가 심하였고 재현성이 결여되었다.

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