• Title/Summary/Keyword: in-vitro

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THE EFFECT OF WASHING PHOSPHORIC ACID ETCHANT ON SHEAR BOND STRENGTH OF AN ORTHODONTIC ADHESIVE (인산 부식액의 수세가 교정용 접착레진의 전단결합강도에 미치는 영향)

  • Kim, Hee-Kyun;Lee, Ki-Soo;Park, Young-Guk
    • The korean journal of orthodontics
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    • v.26 no.5 s.58
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    • pp.497-507
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    • 1996
  • The aim of present study in vitro was to evaluate and compare the effects of different washing times of enamels etched with low phosphoric acid solution which makes unsoluble salts and etched but contaminated with saliva on shear bond strength of an orthodontic adhesive to enamel, and to observe the washing effect on the etched enamel surface by scanning electron microscope. All brackets were bonded with Mono-$Lok2^{TM)}$) on the labial surface of extracted human bicuspids after etching with $20w/w\%\;and\;37w/w$ and phosphoric acid solution for 60seconds and then washing for 0,5,10 and 20seconds respectedly. After etching with $37w/w\%$ phosphoric acid solution and contaminating with saliva for 30seconds and then washing for 0,5,20 and 30seconds and re-etching for 10seconds. After 24hours passed in the $37^{\circ}C$ water bath, the shear bond strengths were measured on Universal Test Machine. The data were evaluated and tested by ANOVA and Duncan's multiple range test, and those results were as follows. 1. There was no significant differences between (p>0.05) shear bond strength of bonded brackets with 5, 10, 20seconds washing etched enamel using $37{\%}w/w{\%}$ phosphoric acid solution. 2. The shear bond strength of bonded brackets with $20w/w\%$ phosphoric acid and then washing for 5seconds showed bonded strength durable to occlusal force but its coefficiency score was high and etched surface was not cleaned completely and therefore it was assumed that its clinical application is not applicable. 3. There was no significant differences between (p>0.05) shear bond strengths of bonded brckets with washing for 5seconds etched enamel using $37w/w\%$ phosphoric acid solution and 10,20 seconds washing etched enamel using $20w/w\%$ phosphoric acid solution. 4. The shear bond strength of washing for 5seconds etched enamel which was contaminated with saliva showed sufficient bonded strength durable to occlusal force but its coefficiency score was high and therefore its clinical application was not applicable. 5. After etching, the sample contaminated with saliva showed the sufficient shear bond strength even washing 20seconds without re-etching.

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Elimination of Apple stem grooving virus from 'Mansoo' pear (Pyrus pyrifolia L.) by an antiviral agent combined with shoot tip culture (항바이러스제 처리와 경정배양에 의한 배(Pyrus pyrifolia L.) '만수'의 Apple stem grooving virus 무병화)

  • Cho, Kang Hee;Shin, Juhee;Kim, Dae-Hyun;Park, Seo Jun;Kim, Se Hee;Chun, Jae An;Kim, Mi Young;Han, Jeom Hwa;Lee, Han Chan
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.391-396
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    • 2016
  • In this study, in vitro-cultured 'Mansoo' pear (Pyrus pyrifolia L.) plants infected with Apple stem grooving virus (ASGV) were used for testing the efficiency of the virus elimination methods. The shoot tips cut from infected plants were treated by thermotherapy ($37^{\circ}C$), cold therapy ($4^{\circ}C$), chemotherapy with ribavirin, and combination of these methods. Treatment periods were 2, 4, and 8 weeks, and concentrations of ribavirin were 20 and $40mg{\cdot}L^{-1}$. The efficiency of ASGV elimination was evaluated by reverse transcription polymerase chain reaction. The shoot survival rate was the highest at 100% after cold therapy, chemotherapy, and combination of two methods, while the rate was the lowest at 33.3% after thermotherapy for 2 weeks. The shoot survival rate after chemotherapy decreased gradually as the treatment period was prolonged. The ASGV elimination rate was the highest at 100% after ribavirin treatment at a concentration of $40mg{\cdot}L^{-1}$ and combination of ribavirin treatment and thermotherapy for 2 weeks, whereas the ASGV elimination rate after cold therapy was the lowest at 16.7%. However, the efficiency of ASGV elimination was enhanced up to 43.3% by the combination of cold therapy and ribavirin treatment. The efficiency of ASGV elimination for all treatments was increased as the treatment period was prolonged. Based on these results, we suggest that ribavirin treatment at a concentration of $20mg{\cdot}L^{-1}$ for 4 weeks or at a concentration of $40mg{\cdot}L^{-1}$ for 2 weeks combined with shoot tip culture was efficient for the elimination of ASGV from pear.

Effect of Digestive Enzymes on the Allergenicity of Autoclaved Market Pork Sausages (가압가열 처리한 시판 돈육 소시지의 항원성에 미치는 소화효소의 영향)

  • Kim, Seo-Jin;Kim, Koth-Bong-Woo-Ri;Song, Eu-Jin;Lee, So-Young;Yoon, So-Young;Lee, So-Jeong;Lee, Chung-Jo;Ahn, Dong-Hyun
    • Food Science of Animal Resources
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    • v.29 no.2
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    • pp.238-244
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    • 2009
  • Food allergy is a serious nutritional problem in both children and adults. Therefore, food allergenicity reduction methods are greatly needed. The allergenicity is altered by various manufacturing processes, and the digestibility of food proteins can be affected by food processing. This study was conducted to investigate the effect of in-vitro digestibility on the allergenicity of autoclaved market pork sausages using pepsin (30min) and trypsin (5, 30, 60, 90, and 120min). The binding ability of the porcine serum albumin (PSA) from sausages A and B significantly decreased by about 30 and 23%, respectively, after autoclave treatment (121; 5, 10, and 30 min). After the pepsin and trypsin treatments, the binding ability of products A and B at 30 min decreased. These competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) results corresponded well with the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting results. The results demonstrated that the allergenicity of pork sausages considerably decreased after autoclave treatment, and were also maintained or decreased after enzyme treatment. Accordingly, autoclave treatment represents a promising processing technology for the reduction of the allergenicity of diverse food products.

A Study on the In-Vitro Test of Thrombogenesis in Centrifugal Biopump (원심성 바이오 펌프를 이용한 혈전 생성의 생체 외 검사에 관한 연구)

  • Na, Myung-Hoon;Kim, Won-Gon;Kim, Joo-Hyun;Kim, Mee-Hyung;Yu, Jae-Hyeon;Lim, Seung-Pyung;Lee, Young
    • Journal of Chest Surgery
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    • v.32 no.10
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    • pp.863-873
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    • 1999
  • 배경: 혈전 생성은 심혈관계 삽입물이나 순환 보조장치의 발전에 있어서 중요한 문제이므로 이러한 장치들의 혈전성에 대한 평가는 필수적이다. 따라서 이러한 장치들의 혈전성 연구를 시행하기 위한 효과적인 생체 외 실험 방식이 개발되어 원심성 순환 펌프의 발전에 큰 도움이 되고 있으며, 최근 엄밀하게 혈액과 공기의 접촉을 차단하는 방식이 믿을 수 있는 생체 외 실험 방식으로 강조되고 있다. 본 논문에서는 모의 순환 회로를 이용하여 기존에 행해진 연구 방법 상의 공기 접촉에 따른 핼액 응고 기전의 활성화 여부를 알아보기 위하여 공기 접촉 여부에 따른 혈액 인자의 변화를 관찰하여 확인하고, 또한 회로 내에서 생성된 혈전을 생체 내에서 형성된 혈전과 비교 분석하여 이 모의 순환 회로를 이용한 생체 외 검사가 새로 고안된 심혈관계 장치들의 혈전성을 평가하는데 있어서 동물실험을 대신할 수 있다는 기존의 주장을 검증하고자 하였다. 연구재료 및 방법: 바이오펌프를 이용한 같은 모의 순환 회로 한 쌍을 준비하고 동일 개체에서 얻은 헤파린을 첨가한(1u/$m\ell$) 신선한 혈액을 공기와 접촉시키지 않도록 한 A-회로와 공기와 접촉시킨 B-회로에 충전시켜 실험을 동시에 진행하여 결과를 얻었으며 총 12번 시행하였다. 회로에 충전한 혈액의 activated clotting time(ACT)은 정상의 3~5배였으며 ACT가 정상 범위의 1.5배가되면 실험을 끝냈고, Hematocrit(HCT), 혈소판, 동맥혈가스분석(ABGA), factor Ⅷ, factor \ulcorner, 섬유소원, thromboxane B2(TXB2), free hemoglobin(fHb) 등을 측정하였다. 실험이 끝난 후 A-회로와 B-회로에서 얻은 각 검사를 평가하고 분석하였으며 생성된 혈전을 생체 내에서 생성된 혈전과 조직 검사로 비교하였다. 본 연구의 자료분석은 SPSS 통계 프로그램을 이용하였고 유의수준 0.05를 기준으로 유의도를 판단하였다. 결과: 정상 ACT는 186.9$\pm$20.5초(평균$\pm$표준편차)이었고, 헤파린을 넣은 초기 ACT는 982.2$\pm$165.5초이었으며, 실험시간은 보통 60분에서 180분 사이였다. HCT, fHb, 혈소판, TXB2, factor Ⅷ, factor \ulcorner, 섬유원소의 초기값은 35.5$\pm$3.2%, 12.4$\pm$6.5 mg/dL, 354$\pm$56($\times$103)/$\mu$L, 72.6$\pm$15.1 mg/dL, 29.3$\pm$1.2%, 137.9$\pm$42.1 mg/dL, 그리고 17.7$\pm$9.7%이었다. 공기 접촉에 따른 차이를 보았을 때 ACT(p=0.398), HCT(p=0.988), fHb(p=0.898), 혈소판 (p=0.904), TXB2(p=0.985), factor Ⅷ(p=0.872), factor \ulcorner(p=0.489), 섬유원소(p=0.973)으로 전 예에서 통계적 유의성이 없었다. 실험 후 양 군에서 생성된 혈전의 현미경 소견은 B-회로의 혈전에 여러 군데 공기 방울이 관찰되는 것 이외에 A-회로에서 생성된 혈전과 차이가 없었고, 생체에서 생성된 혈전과 비교했을 때 그 양상이 같았다. A-회로와 B-회로에서 생성된 혈전의 총 량은 8.4$\pm$3.7 mL와 9.4$\pm$3.1 mL로 통계상 유의성은 없었다. (p=0.624). 결론: 모의 순환 회로를 이용한 혈전성 평가를 위한 생체 외 실험에 있어서 공기와의 접촉 여부는 혈액 응고 기전의 활성화나 혈전 생성에 별다른 영향을 미치지 않는다. 모의 순환 회로에서 형성된 혈전과 문헌에 있는 인체 내에서 형성된 혈전을 비교한 결과 그 형태가 일치하므로 이 모의 순환회로를 이용한 실험이 동물실험을 대신할 수 있다고 판단된다.

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Identification of Antagonistic Bacteria, Pseudomonas aurantiaca YC4963 to Colletotri­chum orbiculare Causing Anthracnose of Cucumber and Production of the Antibiotic Phenazine-l-carboxylic acid (Colletotrichum orbiculare에 대한 길항세균 Pseudomonas aurantiaca YC4963의 분리 동정 및 항균물질 Phenazine-1-carboxylic acid의 생산)

  • Chae Hee-Jung;Kim Rumi;Moon Surk-Sik;Ahn Jong-Woong;Chung Young-Ryun
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.342-347
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    • 2004
  • A bacterial strain YC4963 with antifungal activity against Colletotrichum orbiculare, a causal organism of cucumber anthracnose was isolated from the rhizosphere soil of Siegesbeckia pubescens Makino in Korea. Based on physiological and biochemical characteristics and 16S ribosomal DNA sequence analysis, the bac­terial strain was identified as Pseudomonas aurantiaca. The bacteria also inhibited mycelial growth of several plant fungal pathogens such as Botrytis cinerea, Fusarium oxysporum and Rhizoctonia solani on PDA and 0.1 TSA media. The antifungal activity was found from the culture filtrate of this isolate and the active compound was quantitatively bound to XAD adsorption resin. The antibiotic compound was purified and identified as phenazine-l-carboxylic acid on the basis of combined spectral and chemical analyses data. This is the first report on the production of phenazine-l-carboxylic acid by Pseudomonas aurantiaca.

식품과 알레르기: 유전자 재조합 식품의 알레르기 위험성

  • 손대열
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 2000.12a
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    • pp.29-34
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    • 2000
  • 산업 발달에 따라 날로 많은 식품들이 새롭게 개발되어지고 있다. 또한 이와 병행해서 식품으로 인한 알레르기 발생 빈도도 날로 증가하고 있으며 그 증상 또한 점차 심화되고 있는 것이 세계적인 추세이다. 우리나라도 예외는 아니어서 일반 알레르기 환자뿐 아니라 식품으로 인한 알레르기 환자들이 점차 증가됨이 보고되어지고 있다. 농산물 시장의 수입개방이후 우리나라에는 많은 해외 농산물이 수입되어지고 있으며 그 중 작년 한해의 경우 총 수입 농산물의 10%를 넘는 유전자 재조합 농산물이 우리나라에 수입되어진 것으로 통계 보고되어졌다. 이러한 관점에서 알레르기 환자의 증가와 새로운 식품 (특히 유전자 재조합 식품)의 증가에는 서로 관련성이 있을 것으로 추측되어지고 있어 (새로운) 식품에 대한 알레르기성의 예측과 관리가 필요한 실정이다. 이에 몇몇 발표된 유전자 재조합 식품에 관련된 알레르기성 검사 논문들과 실험실에서 이루어진 연구 결과들을 중심으로 유전자 재조합 식품의 알레르기 위험성에 대해 알아보고자 한다. 일반적으로 식품의 단백질이 알레르겐(allergen)으로 작용하기 위해서는 먼저 소화효소에 의해 분해되어지고 장에서 흡수되어져서 immunopotent cell에 의해 process 되어 immune system에 present 되어져야 한다. 따라서 단백질로 인한 알레르기 반응은 그 단백질의 자연적 형태 뿐만이 아니라 소화 효소에 분해된 단편들의 구조 또는 다른 알레르겐 단백질과의 유사 구조로 인한 교차 반응에 의해 발생함을 기억해야 한다. 식품 단백질 중 어떤 단백질이 알레르겐으로 작용하는가에 대한 특이성 조사에 많은 관심이 집중되어지고 있지만 아직까지는 대략 다섯 개 정도의 일반적인 특성으로서 요약되어질 수 있다. 그러나 이러한 대략의 특성에 적용되지 않는 식품 알레르겐도 많음을 잊어서는 안 될 것이다. 알레르겐으로 작용하는 식품 단백질의 일반적 특성 1. 좋은 수용성 2. 식품내에 많은 부분을 차지하는 주 단백질이 주 알레르겐으로 작용 3. 단백질내에 하나 이상의 IgE-binding site 존재 4. 위장액에 대한 저항성 5. 10~70 kDa 크기 유전자 재조합 기술이란 말 그대로 유전자를 인위적으로 새롭게 조합하는 기술로 이전의 기술로는 불가능했던 유전적 변형을 농작물과 동물에 가능하게 했으며 이로 인해 유전적으로 변형된 식용 동식물의 개발이 가능하게 되었다. 새로운 유전인자를 개체에 삽입함으로 새로운 단백질이 발현 될수 있고 그로 인해 1) 해충과 질병에 대한 저항성 증가, 2) 화학 제초제에 대한 새로운 저항성 부여, 3) 식품의 저장성 향상, 4) 식품의 영향적 보충/향상 등의 이점을 얻을 수 있다 (표 1). 세계적으로 유전자 재조합 된 새로운 농산물의 재배는 날로 증가추세에 있으며 그 중에서 가장 많은 부분을 차지하는 농산물로 soybean을 들 수 있으며 (표 2) soybean을 중심으로 그 알레르기성의 변화가 연구 조사된 몇 가지 예를 살펴보고자 한다. (표 3)에 요약된 soybean중 첫 번째 경우는 재초제에 대한 저항성을 높여주기 위해 Agrobacterium에 존재하는 EPSPS라는 단백질을 콩에서 발현하도록 찬 유전자 재조합 된 콩의 경우이다. 이 콩의 경우에는 첫째. 이전된 새로운 단백질 EPSPS가 다른 여러 식물에 이미 존재하고 있는 단백질로서 우리가 이미 이러한 식품을 섭취할 때 이 단백질도 같이 섭취해오고 있었다는 점, 둘째. 이 단백질이 소화액 분해 실험에서 짧은 시간내에 분해가 되었다는 점, 셋째. 재조합 된 콩과 자연 콩이 성분 분석에서 차이를 나타내지 않았다는 점, 네 번째. 쥐를 통한 다양섭취 실험에서 아무런 이상 반응이 없었다는 점등의 결과를 기준으로 알레르기에 대한 개별 검사 없이 안전한 콩으로 결론짓고 있다. 영양성을 높이기 위해 Brazil nut에서 methionine 함량이 풍부한 2s albumine을 콩에서 발현하도록 한 두 번째 유전자 재조합 콩의 경우 이전된 단백질 때문에 Brazil nut에 알레르기 반응을 일으키는 알레르기 환자들을 조사한 결과 역시 재조합 된 콩에도 알레르기 반응을 일으켰다는 보고이다. Brazil nut에서 콩으로 이전된 단백질이 Brazil nut에서의 알레르기성을 그대로 유지한 점을 볼 때 새로운 단백질이 어디에서 유래하는가가 중요함을 잘 보여준 연구이다 세 번째 콩의 경우 역시 영양성을 높여주기 위해 corn에서 10 kDa과 HSZ 단백질을 콩에서 발현하도록 유전자 재조합했는데 이 콩의 경우는 알레르기 환자들이 유전자 재조합 된 콩과 자연 콩에 반응의 차이를 나타내지 않았다는 결과 보고이다. 위의 세 실험 결과들을 종합해 볼 때 무엇보다도 새롭게 발현된 단백질이 원래 어떤 성질을 갖고 있으며 어디에서 유래했는지가 알레르기성 조사에 중요한 역할을 한다 할 수 있겠다. 또한 유전자 재조합된 식품들은 알레르기 환자들을 위해 표기되어져야 할 것인데 이를 위한 알레르기성 검사 실험은 공공단체를 통해 이루어져야 할 것이며 환자들마다 알레르겐으로 작용하는 단백질의 인식부위(epitope)가 다를 수 있기 때문에 적어도 10명 이상의 알레르기 환자들이 조사되어져서 검사가 이루어져야 할 것이다. 환자들의 혈청을 통한 in vitro 실험에서는 ELISA, RAST, immunoblotting과 같은 검사 방법들이 적용될 수 있고, 그 결과가 음성인 경우에 그 다음 단계로 in vivo 실험에서는 직접 환자의 피부반응검사 (skin prick test)나 DBPCFC (double-blind placebo-controlled food challenge) 검사 방법을 통해 확인되어져서 이 모든 경우가 음성인 경우와 하나라도 양성인 경우를 구별하여 식품에 표기함으로 알레르기 환자들의 유전자 재조합 식품에 대한 안전성이 보장되어져야 할 것이다.

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Potential Antitumor $\alpha$-Methylene-$\gamma$-butyrolactone-Bearing Nucleic Acid Base. 3. Synthesis of $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans

  • Kim, Jack-C.;Kim, Si-Hwan;Kim, Ji-A;Choi, Soon-Kyu;Park, Won-Woo
    • Archives of Pharmacal Research
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    • v.21 no.4
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    • pp.458-464
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    • 1998
  • Search for a new $\alpha$-methylene-$\gamma$-butyrolactone-bearing 6-substituted purine as a potental antitumor agent has led to synthesize seven, hitherto unreported, $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$- methylenetetrahydrofurans (H, Cl, l, $CH_3$, $NH_2$, SH, >C=O) (6a-g). These include $5^1$-Methyl-$5^1$-[(9H-purin-9-yl)methyll-$2^1$-oxo-$3^1$ -methylenetetrahydrofurans (6a), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydr ofurans (6b), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6c), $5^1$-Methyl-$5^1$-[(6-methyl-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6d), $5^1$-Methyl-$5^1$-[(9H-adenin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6e), $5^1$-Methyl-$5^1$-[(6-mercapto-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6f) and $5^1$-Methyl-$5^1$-[(9H-hypoxanthin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrof urans (6g) which were made by the Reformatsky-type reaction of ethyl $\alpha$-(bromomethyl) acrylate with the corresponding (6-substituted-9H-purin-9-yl)-2-propanone intermediates (5a-g). These ketone intermediates 5a-g, 1-(9H-purin-9-yl)-2-propanone (5a), 1-(6-chloro-9H-purin-9-yl)-2-propanone (5b), 1-(6-iodo-9H-purin-9-yi)-2-propanone (5c), 1-(6-methyl-9H-purin-9-yl)-2-propanone (5d), 1-(9H-adenin-9-yl)-2-propanone (Se), 1-(6-mercapto-9H-purin-9-yl)-2-propanone (5f), and 1-(9H-hypoxanthin-9-yl)-2-propanone (5g) were directly obtained by the alkylation of the 6-substituted purine bases with the chloroacetone in the presence of $K_2$$CO_3$ (or NaH) under DMF (or DMSO). The preliminary in vitro cytotoxcity assay for the synthetic .alpha.-methylene-y-butyro-lactone compounds (6a-g) were determined against three cell lines (PM-3A, P-388, and K-562) and showed the moderate antitumor activity ($IC_50$ ranged from 1.4 to 4.3 $\mu\textrm{g}$/ml) with the compound $5^1$-methyl-$5^1$ -[(9H-hypoxanthin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofuran (6g) showing the least antitumor activity.

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Nutritional Composition and Cytoprotective Effect of Moringa oleifera Lam. (Drumstick-tree (Moringa oleifera Lam.)의 주요 영양성분 및 추출물의 신경세포 보호 효과)

  • Jin, Su Il;Kim, Hyeon Ju;Jeong, Ji Hee;Jin, Dong Eun;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.46 no.5
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    • pp.609-615
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    • 2014
  • The cytoprotective effect of Moringa oleifera Lam. (drumstick tree) on neuronal cells was investigated to confirm the physiological benefits associated with this natural food resource. First, the drumstick tree extract was chemically analyzed to determine inherent nutritional constituents. Calcium and potassium were identified as the major mineral constituents, and palmitic acid (C16:0, 16.33%) and gadoleic acid (C20:01, 66.34%) were detected as the major fatty acids. Moreover, drumstick tree extract contained 94.78 mg/100 g vitamin E and 112.61 mg/100 g niacin. PC12 cells were used to study the cytoprotective effects of drumstick tree extract. Intracellular accumulation of reactive oxygen species was significantly reduced when $H_2O_2$ treated-neuronal cells were cultured in a medium containing the methanolic extract of drumstick tree, compared to cells treated with only $H_2O_2$. Cell viability assay using MTT showed that the extract protected cells against $H_2O_2$-induced neurotoxicity and inhibited LDH leakage from the cell membrane. Caspase assay showed that the extract exerted cytoprotective effect against apoptosis. Consequently, these data suggest that drumstick tree is a useful natural resource with positive effects on human health.

The Role of Protein Kinase C in Acute Lung Injury Induced by Endotoxin (내독소에 의한 급성폐손상에서 Protein Kinase C의 역할)

  • Kim, Yong-Hoon;Moon, Seung-Hyug;Kee, Sin-Young;Ju, Jae-Hak;Park, Tae-Eung;Im, Keon-Il;Cheong, Seung-Whan;Kim, Hyeon-Tae;Park, Choon-Sik;Jin, Byung-Won
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.2
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    • pp.349-359
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    • 1997
  • Background : The signal pathways and their precise roles for acute respiratory distress syndrome caused by endotoxin (ETX) has not been established. Since there has been several in vitro experiments suggesting that activation of protein kinase C (PKC) pathway may be responsible for endotoxin-induced inflammatory reaction, we performed in vivo experiments in the rats with the hypothesis that PKC-inhibition can effectively prevent endotoxin-induced acute lung injury. Methods : We studied the role of PKC in ETX-induced ALI using PKC inhibitor (staurosporine, STP) in the rat Specific pathogen free male Sprague-Dawley weighted from 165 to 270g were used for the study. Animals were divided into the normal control (NC)-, vehicle control (VC)-, ETX-, PMA (phorbolmyristateacetate)-, STP+PMA-, and STP+ETX-group. PMA (50mg/kg) or ETX (7mg/kg) was instilled through polyethylen catheter after aseptic tracheostomy with and without STP (0.2mg/kg)-pretreatment STP was injected via tail vein 30min before intratracheal injection (IT) of PMA or ETX. Bronchoalveolar lavage (BAL) was done 3-or 6-hrs after IT of PMA or ETX respectively, to measure protein concentration, total and differential cell counts. Results : The results were as follows. The protein concentrations in BALF in the PMA- and ETX-group were very higher than that of VC-group (p<0.001). When animals were pretreated with STP, the %reduction of the protein concentration in BALF was $64.8{\pm}8.5$ and $30.4{\pm}2.5%$ in the STP+PMA- and STP+ETX-group, respectively (p = 0.028). There was no difference in the total cell counts between the PMA-and VC-group (p = 0.26). However the ETX-group showed markedly increased total cell counts as compared to the VC- (p = 0.003) and PMA-group (p = 0.0027), respectively. The total cell counts in BALF were not changed after pretreatment with STP compared to the PMA- (p = 0.22) and ETX-group (p = 0.46). The percentage of PMN, but not alveolar macrophage, was significantly elevated in the PMA-, and ETX-group. Especially in the ETX-group, the percentage of PMN was 17 times higher than that of PMA (p < 0.001). The differential cell counts was not different between the PMA and STP+PMA On the contrary the STP+ETX-group showed decreased percentage of PMN (p = 0.016). There was no significant relationship between the protein concentration and the total or differential cell counts in each group. Conclusion : Pretreatment with PKC-inhibitor (staurosporine) partially but significantly inhibited ETX-induced ALI.

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Plasma Activity of Lysosomal Enzymes in Active Pulmonary Tuberculosis (활동성 폐결핵 환자에서 혈중 리소솜 효소의 활성도)

  • Koh, Youn-Suck;Choi, Jeong-Eun;Kim, Mi-Kyung;Lim, Chae-Man;Kim, Woo-Sung;Chi, Hyun-Sook;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.5
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    • pp.646-653
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    • 1995
  • Background: The confirmative diagnosis of pulmonary tuberculosis(Tb) can be made by the isolation of Mycobacterium Tuberculosis(MTb) in the culture of the sputum, respiratory secretions or tissues of the patients, but positive result could not always be obtained in pulmonary Tb cases. Although there are many indirect ways of the diagnosis of Tb, clinicians still experience the difficulty in the diagnosis of Tb because each method has its own limitation. Therefore development of a new diagnostic tool is clinically urgent. It was reported that silica cause some lysosomal enzymes to be released from macrophages in vitro and one of these enzymes is elevated in workers exposed to silica dust and in silicotic subjects. In pulmonary Tb, alveolar macrophages are known to be activated after ingestion of MTb. Activated macrophages can kill MTb through oxygen free radical species and digestive enzymes of lysosome. But if macrophages allow the bacilli to grow intracellularly, the macrophages will die finally and local lesion will enlarge. Then it is assumed that the lysosomal enzymes would be released from the dead macrophages. The goal of this investigation was to determine if there are differences in the plasma activities of lysosomal enzymes, ($\beta$-glucuronidase(GLU) and $\beta$-N-acetyl glucosaminidase(NAG), among the groups of active and inactive pulmonary Tb and healthy control, and to see if there is any possibility that the plasma activity of GLU and NAG can be used as diagnostic indicies of active pulmonary Tb. Methods: The plasma were obtained from 20 patients with bacteriologically proven active pulmonary Tb, 15 persons with inactive Tb and 20 normal controls. In 10 patients with active pulmonary Tb, serial samples after 2 months of anti-Tb medications were obtained. Plasma GLU and NAG activities were measured by the fluorometric methods using 4-methylumbelliferyl substrates. All data are expressed as the mean $\pm$ the standard error of the mean. Results: The activites of GLU and NAG in plasma of the patients with active Tb were $21.52{\pm}3.01$ and $325.4{\pm}23.37$(nmol product/h/ml of plasma), respectively. Those of inactive pulmonary Tb were $24.87{\pm}3.78$, $362.36{\pm}33.92$ and those of healthy control were $25.45{\pm}4.05$, $324.44{\pm}28.66$(nmol product/h/ml of plasma), respectively. There were no significant differences in the plasma activities of both enzymes among 3 groups. The plasma activities of GLU at 2 months after anti-Tb medications were increased($42.18{\pm}5.94$ nmol product/h/ml of plasma) in the patients with active pulmonary Tb compared with that at the diagnosis of Tb(P-value <0.05). Conclusion: The results of the present investigation suggest that the measurement of the plasma activities of GLU and NAG in the patients with active pulmonary Tb could not be a useful method for the diagnosis of active Tb. Further investigation is necessary to define the reasons why the plasma activities of the GLU was increased in the patients with active pulmonary Tb after Tb therapy.

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