• Food Science and Biotechnology
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• 제14권3호
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• pp.399-404
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• 2005

The aim of this study was to investigate the in vitro antioxidant profiles of genistein and other isoflavonoids. The reactivity of genistein towards stable radical and reactive oxygen species including ${\bullet}\;ABTS^+$, ${\bullet}{O_2}^-$, $H_2O_2$ and HOCl has been investigated, and the effects were compared with other isoflavonoids and antioxidants. All the tested isoflavonoids showed remarkable ${\bullet}\;ABTS^+$ scavenging activity and genistein was more potent than BHT and ascorbic acid. Genistein was more effective in scavenging hypochlorous acid than superoxide and hydrogen peroxide. At $10\;{\mu}M$ concentrations of genistein and genistin showed about 90% inhibitory effect on HOCl, while BHT and ascorbic acid showed lower than 50% inhibitory effect. Moreover, genistein could inhibit plasmid DNA cleavage, protein degradation and cell death from HOCl attack, while daidzein, BHT and ascorbic acid could not protect them effectively. These results suggest that genistein is a more potent radical scavenger than other isoflavonoids, and it can remarkably reduce cellular damage induced by HOCl.

• 생명과학회지
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• 제22권9호
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• pp.1187-1193
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• 2012

본 연구는 할로겐 화합물의 첨가가 in vitro 상의 반추위 발효성상과 메탄생성에 미치는 영향에 대한 효과를 규명하고자 실시하였다. Italian rye grass 및 배합사료를 6:4의 비율로 급여한 반추위 cannula가 시술된 홀스타인에서 반추위액을 채취하여 사용하였고, 채취된 반추위액은 분쇄된 timothy (대조구; C)에 bromochloromethane (BCM구), 2-Bromoethanesulfonic acid (BES구), 3-Bromopropanesulfonic acid (BPS구), chloroform (CLF구) 및 Pyromellitic diimide (PMDI구)의 5가지 할로겐 화합물을 각각 1 ppm씩 첨가하여 in vitro 배양하였다. pH는 6.72에서 6.25 정도로 배양시간이 경과함에 따라 낮아지는 경향을 나타내었고, 배양 48시간에는 처리구간 차이가 없었다. 배양 48시간 후의 총 가스 발생량은 처리구간 유의적인(p<0.05) 차이는 없었고, BPS구를 제외한 모든 처리구에서의 메탄 발생량은 대조구에 비해 유의적으로(p<0.05) 감소하였다. 배양 12시간에서의 총 휘발성 지방산 및 propionic acid의 발생량은 처리구가 대조구에 비해 유의적으로(p<0.05) 높았다. 본 실험의 결과, 할로겐 화합물의 첨가는 반추위 내의 pH, 건물 소화율, 미생물 수 및 총 가스 발생량의 발효 성상에는 영향을 주지 않으면서 메탄의 발생량이 감소 되었다. 실험 결과를 종합해 보면, 할로겐 화합물 첨가는 반추위 내 pH, 가스 발생량, 반추위 미생물 성장량 및 propionic acid 모두 증가하였으며, 반추위내 메탄생성을 억제하였다. 앞으로 할로겐화합물과 다른 메탄억제 물질과 혼합하여 반추위 내 메탄생성 억제에 관한 구체적인 연구가 필요한 것으로 생각된다.

• Tuberculosis and Respiratory Diseases
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• 제54권4호
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• pp.449-458
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• 2003

연구배경 : Cyclosporin A(CsA)와 tacrolimus(FK506)은 현재 임상에서 널리 쓰이는 면역억제제이다. CsA와 FK506이 $I{\kappa}B/NF-{\kappa}B$ 경로에 미치는 영향에는 세포에 따라 다양한 효과가 알려져 있다. 그러나 CsA와 FK506이 기관지 상피세포에서 $I{\kappa}B/NF-{\kappa}B$ 경로에 미치는 효과에 관해서는 알려져 있지 않고, 각종 염증 세포에서의 차이에 관해서도 보고가 미미한 실정이다. 본 연구에서는 비염증세포인 기관지상피세포와, 폐의 염증에 중요한 역할을 하는 염증 세포(폐포대식세포, 단핵구, 림프구)에서 CsA와 FK506이 $I{\kappa}B{\alpha}$의 분해에 미치는 영향과 그 기전을 평가하였다. 방 법 : 비염증세포로는 BEAS-2B와 A549 세포주를 이용하였다. FK506 또는 CsA 전처치 후 TNF-${\alpha}$로 자극하고 anti-$I{\kappa}B{\alpha}$ 항체를 이용한 Western blot으로 $I{\kappa}B{\alpha}$의 분해 여부를 관찰하였다. 염증세포로는 폐포대식세포, 말초혈액 단핵구 및 림프구를 이용하였고, 역시 FK506 또는 CsA 전처치 후 TNF-${\alpha}$, IL-$1{\beta}$, LPS로 자극하고 anti-$I{\kappa}B{\alpha}$ 항체를 이용한 Western blot으로 $I{\kappa}B{\alpha}$의 분해 여부를 관찰하였다. IKK의 활성도는 GST-$I{\kappa}B{\alpha}$를 기질로 이용한 in vitro immune complex kinase assay로 평가하였다. 결 과 : 사용된 모든 세포에서 CsA와 FK506은 $I{\kappa}B{\alpha}$의 발현에 영향을 미치지 않았다. 기관지 상피세포에서 TNF-${\alpha}$ 자극에 의한 $I{\kappa}B{\alpha}$의 분해는 CsA의 전처치로 억제되었으나, FK506의 전처치로는 억제되지 않았다. 단핵구, 림프구 및 폐포대식세포에서 외부자극에 의한 $I{\kappa}B{\alpha}$의 분해는 CsA 또는 FK506의 전처치로 억제되었으나 IKK활성은 억제되지 않았다. 결 론 : CsA와 FK506은 각각 기관지 상피세포와 단핵구, 림프구, 폐포대식세포에서 외부 자극에 의한 $I{\kappa}B{\alpha}$의 분해를 억제하는데, 이는 IKK 활성화의 억제가 아닌 다른 경로를 통하는 것으로 생각된다.

• 산업식품공학
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• 제21권3호
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• pp.286-291
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• 2017

약용식물의 열수 추출물이 in vitro에서 alcohol dehydrogenase (ADH)와 aldehyde dehydrogenase (ALDH)의 활성에 미치는 영향을 확인하였다. 약용식물에 20배의 증류수를 넣고 $80^{\circ}C$에서 8시간 추출하여 얻어진 추출액을 시료로 사용하였다. 50종의 약용식물 중에서 마늘과 육계 추출물이 숙취해소 천연소재로서의 활용 가능성이 가장 높게 나타났다. 마늘 추출물은 ADH에 비해 ALDH의 활성을 2배 이상 촉진시킴으로써 acetaldehyde의 분해가 잘 되게 하였다. 육계 추출물은 ALDH의 활성에 비해 ADH의 활성을 획기적으로 저해함으로써 acetaldehyde의 생성을 크게 억제하였다. 육계 추출물은 농도에 비례하여 ADH와 ALDH의 활성을 저해하였으며, $45.33{\mu}g/mL$의 농도에서 ADH의 활성을 52.8% 저해하였고 ALDH의 활성을 11.0% 저해하였다.

• 생약학회지
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• 제38권2호통권149호
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• pp.101-107
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• 2007

This study was carried out to examine the in vitro effects of alginate (LVA) which is low viscosity alginic acid, on collagen type II synthesis of chondrocytes and the in vivo effect, orally administered, on cartilage degradation. Rabbit articular chondrocytes were cultured in 35 mm dishes and then LVA was treated. The effects of LVA of various viscosity (86.5 cP(LVA1),45.4 cP(LVA2), 21.2 cP(LVA3) and 9.6 cP(LVA4)) and various concentration (50, 100, 200 ${\mu}$g/ml of LVA4) on chondrocytes were determined by western blotting assay for the detection of collagen type II production. In western blotting assay, collagen type II production in chondrocytes were 1.00 in control,0.95 in LVA1, 1.41 in LVA2, 1.57 in LVA3 and 1.58 in LVA4. Collagen type II production of various concentration of LVA4 were 1.00 in control, 1.24 in 50 ${\mu}$g/ml of LVA4, 1.52 in 100 ${\mu}$g/ml of LVA4 and 1.86 in 200 ${\mu}$g/ml of LVA4. Osteoarthritis (OA) was induced in 24 rabbits by unilateral anterior cruciate ligament transection (ACLT) and randomly divided into 6 groups. The experimental group was given oral administration of 0.5 ml/kg of saline(control), 12.5 mg/kg of LVA (A12.5), 25 mg/kg of LVA (A25), 50 mg/kg of LVA (A5O), 75 mg/kg of LVA (A75) and 20 mg/kg of aceclofenac (AC) for 6 weeks after ACLT. All knees were harvested at 6 weeks after surgery and cartilage degradation was evaluated. Cartilage degradation in the control group was significantly more severe than that in the A25, A5O and A75 groups but AC group had no significant changes on the macroscopic grading scale.

• Animal Bioscience
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• 제35권11호
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• pp.1733-1743
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• 2022

Objective: This study was aimed to explore the efficacy of combination of endo-xylanase (Xyn) and xylan-debranching enzymes (arabinofuranosidase, Afd and feruloyl esterase, FE) in improving utilization of bran in piglet diet. Methods: In vitro experiments were firstly conducted to examine the enzymological properties of Xyn, Afd, and FE, concurrent with their effect on degradation of arabinoxylan (Abx) in bran. In vivo experiment was then implemented by allocating two hundred and seventy 35-d-old postweaning piglets into 3 groups (6 replicates/group), which received bran-containing diet supplemented with Xyn (1,600 U/kg) or its combination with Afd (0.8 U/kg) and FE (4 U/kg) or without enzyme. Results: Both Xyn, Afd, and FE are relatively stable against the changes in temperature and pH value. Combining Xyn with Afd and FE had a superiority (p<0.05) over Xyn alone and its combination with Afd or FE in promoting (p<0.05) degradation of Abx in different brans. Combined treatment with Xyn, Afd, and FE was more beneficial than Xyn alone to induce increasing trends (p<0.10) of average daily gain, final body weight and feed efficiency of piglets fed bran-containing diet. Moreover, combination of Xyn, Afd, and FE showed advantages (p<0.05) over Xyn alone in causing reductions (p<0.05) in diarrhea rate and cecal pH value, concurrent with increases (p<0.05) in cecal and colonic acetic acid and total volatile fatty acid concentrations, as well as cecal butyric acid concentration of piglets fed bran-containing diet. Conclusion: Combining Xyn with Afd and FE was more beneficial than Xyn alone in promoting degradation of Abx in bran, along with growth performance and intestinal volatile fatty acid profile of piglets received bran-containing diet. Thereby, combination of Xyn, Afd, and FE had a superior efficacy relative to Xyn alone in improving application of cereal bran in piglet diet.

• 대한본초학회지
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• 제23권3호
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• pp.127-134
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• 2008

Objectives : The present study was designed to investigate whether Ostericum koreanum (OK) could regulate lipopolysaccharide (LPS)-induced inflammatory response in vitro and in vivo. Methods : To evaluate of anti-inflammatory effect of OK, we examined Nitric oxide (NO), proinflammatory cytokines production in LPS-stimulated RAW264.7 cells. Furthermore, we checked molecular mechanism especially in the phosphorylation of mitogen-activated protein kinases (MAPKs) and the degradation of inhibitory kappa B a ($Ik-B{\alpha}$) using western blot and also investigated survival of mice in LPS-mediated endotoxin shock. Results : 1. Extract from OK itself have weak cytotoxic effect on RAW264.7 cells. Extract from OK inhibited LPS-induced NO, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, IL-6 and IL-10 production in RAW264.7 cells. 2. OK inhibited the phosphorylation of MAPKs, such as p38, extracelluar signal-regulated kinase (ERK1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of $I{\kappa}-B{\alpha}$ in the LPS-stimulated RAW264.7 cells 3. OK did not inhibit LPS-induced endotoxin shock. Conclusions : OK down-regulated LPS-induced NO and cytokines production through suppressing activation of MAPKs and degradation of $I{\kappa}-B{\alpha}$. Our results suggested that OK may be a beneficial drug against inflammatory diseases.

• BMB Reports
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• 제40권5호
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• pp.670-677
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• 2007

Although polysaccharide is believed to play an important role in the medicinal effect of Dendrobium chrysotoxum Lindl (DCL), its role as an antioxidant and in anti-hyperglycemic induction was not reported. In this study, polysaccharide with molecular weight of approximately 150 kDa, herein named DCLP, was isolated from the stem of DCL, and its antioxidative, hypoglycemic and immune stimulating effects were evaluated using various in vitro and in vivo assay systems. DCLP inhibited hydroxyl radicals ($^{\cdot}$OH)-mediated deoxyribose degradation by scavenging hydroxyl radicals directly as well as by chelating iron ions. DCLP also showed dose-dependent scavenging activity on superoxide anions ($O_2^{{\cdot}-}$) and offered significant protection (p < 0.001) against glucose oxidase-mediated cytotoxicity in Jurkat cells. DCLP had immune stimulating effects, as evidenced by the DCLP-mediated increases in the level of DNA synthesis, viability, and cytokine secretion in mouse lymphocytes. Moreover, oral administration of DCLP produced a significant reduction in blood glucose level in alloxan-induced diabetic mice. These findings suggest that DCLP has a potential utility in treating patients who require enhanced antioxidation, immune function and/or hypoglycemic activity.

• 한국수정란이식학회지
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• 제32권3호
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• pp.105-109
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• 2017

Autophagy is an intracellular degradation and recycling system. Oocyte maturation is dynamic process, in which various proteins should be synthesized and degraded. In our previous study, we reported the loci of autophagosome and dynamics of autophagic activity in porcine oocytes during in vitro maturation. In this study, we verified loci of autophagosome in porcine follicular cumulus-oocyte complex by detection of microtubule-associated protein 1A/1B-light chain 3 (LC3) which is the reliable marker of autophagosome. Porcine ovary including various sizes of follicles was fixed within 1 hour after collection from slaughterhouse. After fixation, immunohistochemistry was conducted on sliced ovary tissue containing various sizes of follicles by using LC3 antibody. As a result, LC3 signal was clearly detected in both cumulus and oocytes of various sizes of follicles. We also found ring shaped signal which represent autophagosome near oocyte membrane. Most of the signals in oocytes were localized nearby cellular membrane while evenly dispersed in cumulus cells. Therefore, this result suggests that autophagy occurs in porcine COCs (cumulus-oocyte complexes) at follicular stage.

• Asian-Australasian Journal of Animal Sciences
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• 제22권3호
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• pp.356-364
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• 2009

The current ruminant feeding models require parameterization of the digestion kinetics of carbohydrate fractions in feed ingredients to estimate the supply of nutrients from a ration. Using an automated gas production technique, statistically welldefined digestion rate of carbohydrate, including soluble carbohydrate, can be estimated in a relatively easy way. In this study, the gas production during in vitro fermentation was measured and recorded by an automated gas production system to investigate degradation kinetics of carbohydrate fractions of a wide range of ruminant feeds: corn silage, rice straw, corn, soybean hull, soybean meal, and cell mass from lysine production (CMLP). The gas production from un-fractionated, ethanol insoluble residue and neutral detergent insoluble residue of the feed samples were obtained. The gas profiles of carbohydrate fractions on the basis of the carbohydrate scheme of the Cornell Net Carbohydrate and Protein System (A, B1, B2, B3 and C) were generated using a subtraction approach. After the gas profiles were plotted with time, a curve was fitted with a single-pool exponential equation with a discrete lag to obtain kinetic parameters that can be used as inputs for modern nutritional models. The fractional degradation rate constants (Kd) of corn silage were 11.6, 25.7, 14.8 and 0.8%/h for un-fractioned, A, B1 and B2 fractions, respectively. The values were statistically well estimated, assessed by high t-value (>12.9). The Kd of carbohydrate fractions in rice straw were 4.8, 21.1, 5.7 and 0.5%/h for un-fractioned, A, B1 and B2 fractions, respectively. Although the Kd of B2 fraction was poorly defined with a t-value of 4.4, the Kd of the other fractions showed tvalues higher than 21.9. The un-fractioned corn showed the highest Kd (18.2%/h) among the feeds tested, and the Kd of A plus B1 fraction was 18.7%/h. Soybean hull had a Kd of 6.0, 29.0, 3.8 and 13.8%/h for un-fractioned, A, B1 and B2, respectively. The large Kd of fraction B2 indicated that NDF in soybean hull was easily degradable. The t-values were higher than 20 except for the B1 fraction (5.7). The estimated Kd of soybean meal was 9.6, 24.3, 5.0%/h for un-fractioned, A and B1 fractions, respectively. A small amount of gas (5.6 ml at 48 ho of incubation) was produced from fermentation of CMLP which contained little carbohydrate. In summary, the automated gas production system was satisfactory for the estimation of well defined (t-value >12) kinetic parameters and Kd of soluble carbohydrate fractions of various feedstuffs that supply mainly carbohydrate. The subtraction approach, however, should be applied with caution for some concentrates, especially those which contain a high level of crude protein since nitrogen-containing compounds can interfere with gas production.