• 동의생리병리학회지
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• 제27권6호
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• pp.802-808
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• 2013

Here we tried to uncover the potential anti-lipogenic effect and the underlying mechanism of Phaseolus angularis seed in a cellular model of nonalcoholic fatty liver disease (NAFLD) induced in HepG2 cells. Ethanol extract of Phaseolus angularis seed (JSD) was prepared. HepG2 cells were incubated in palmitate containing media to induce intracellular lipid accumulation, and co-treated with JSD for 16 hrs before examine intracellular lipid content. In control group, the cells were not co-treated with JSD. We measured the effects of JSD on liver X receptor ${\alpha}$ ($LXR{\alpha}$) and sterol regulatory element-binding transcription factor-1c (SREBP-1c) expression, transcription level of lipogenic genes, including acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD-1), and AMP-activated protein kinase (AMPK) activation in HepG2 cells. JSD markedly reduced palmitate-induced intracellular lipid accumulation in HepG2 cells. JSD suppressed $LXR{\alpha}$/SREBP-1c expression, and SREBP-1c mediated induction of ACC, FAS, and SCD-1. Furthermore, JSD activated AMPK, which plays a major role in the control of hepatic lipid metabolism. Taken together, it is suggested that JSD has a potential to alleviate hepatic steatosis, at least in part, by suppressing $LXR{\alpha}$/SREBP-1c mediated induction of lipogenic genes. In addtion, the anti-lipogenic potential may be associated with activation of AMPK. Therefore, the Phaseolus angularis seed could be applied as a potential therapeutics for NAFLD with additional clinical studies.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.717-724
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• 2003

A newly isolated Citrobacter sp. Y19 catalyzes the CO-dependent $H_2$ production (biological water-gas shift reaction) by the actions of CO dehydrogenase (CODH) and hydrogenase. Y 19 requires $O_2$ for fast growth, but its $H_2$ production activity is significantly inhibited by $O_2$. In the present study, the effect of $O_2$ on the activities of CODH ard hydrogenase was investigated quantitatively in both whole cells and broken cells, based on CO-dependent or methyl viologen (MV)-dependent $H_2$ production in addition to CO-dependent MV reduction. In crude cell extracts, CODH activity was mostly found in the soluble fraction. Inactivation of CODH and hydrogenase activities by $O_2$ followed the first-order decay kinetics, and the dependence of the rate constants on $O_2$ partial pressure could be expressed by the Michaelis-Menten equation. In whole cells, the maximum deactivation rate constants ($k_{d,max}$ of hydrogenase and CODH were quite similar: $0.07{\pm}0.03 min^{-1}\;and\;0.10{\pm}0.04 min^{-1}$, respectively. However, the first-order rate constant ($k_{d,max}/K_s$) of CODH ($0.25\;min^{-1}\;atm^{-1}$) at low $O_2$ partial pressures was about 3-fold higher than that of the hydrogenase, since the half-saturation constant ($K_s$) of CODH was about half of that of hydrogenase. In broken cells, both enzymes became significantly more sensitive to $O_2$ compared to the unbroken cells, while $k_{d,max}/K_s$ increased 37-fold for hydrogenase and 6.7-fold for CODH. When whole cells were incubated under anaerobic conditions after being exposed to air for 1 h, hydrogenase activity was recovered more than 90% in 2 h suggesting that the deactivation of hydrogenase by $O_2$ was reversible. On the contrary, CODH activity was not recovered once deactivated by $O_2$ and the only way to recover the activity was to synthesize new CODH. This study indicates that $O_2$ sensitivity of $H_2$ production activity of Citrobacter sp. Y19 is an important drawback as in other $H_2-producing$ bactria.

• Asian-Australasian Journal of Animal Sciences
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• 제26권2호
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• pp.266-274
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• 2013

The effect of different phytogenic feed additives on reducing odorous compounds in swine was investigated using in vitro fermentation and analyzed their microbial communities. Soybean meal (1%) added with 0.1% different phytogenic feed additives (FA) were in vitro fermented using swine fecal slurries and anaerobically incubated for 12 and 24 h. The phytogenic FAs used were red ginseng barn powder (Panax ginseng C. A. Meyer, FA1), persimmon leaf powder (Diospyros virginiana L., FA2), ginkgo leaf powder (Ginkgo biloba L., FA3), and oregano lippia seed oil extract (Lippia graveolens Kunth, OL, FA4). Total gas production, pH, ammonianitrogen ($NH_3$-N), hydrogen sulfide ($H_2S$), nitrite-nitrogen ($NO_2{^-}$-N), nitrate-nitrogen ($NO_3{^-}$-N), sulfate (${SO_4}^{--}$), volatile fatty acids (VFA) and other metabolites concentration were determined. Microbial communities were also analyzed using 16S rRNA DGGE. Results showed that the pH values on all treatments increased as incubation time became longer except for FA4 where it decreased. Moreover, FA4 incubated for 12 and 24 h was not detected in $NH_3$-N and $H_2S$. Addition of FAs decreased (p<0.05) propionate production but increased (p<0.05) the total VFA production. Ten 16S rRNA DGGE bands were identified which ranged from 96 to 100% identity which were mostly isolated from the intestine. Similarity index showed three clearly different clusters: I (FA2 and FA3), II (Con and FA1), and III (FA4). Dominant bands which were identified closest to Eubacterium limosum (ATCC 8486T), Uncultured bacterium clone PF6641 and Streptococcus lutetiensis (CIP 106849T) were present only in the FA4 treatment group and were not found in other groups. FA4 had a different bacterial diversity compared to control and other treatments and thus explains having lowest odorous compounds. Addition of FA4 to an enriched protein feed source for growing swine may effectively reduce odorous compounds which are typically associated with swine production.

• 대한치과교정학회지
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• 제35권1호
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• pp.51-59
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• 2005

본 연구는 교정환자의 브라켓과 치아 경계부 및 브라켓으로부터 2mm 이상 떨어진 치아 평활면의 치면세균막에 존재하는 mutans streptococci의 종 및 생물형에 차이가 있는지를 알아보고자 시행되었다. 조선대학교 치과병원에 내원한 13세 이상 35세 미만의 환자 28명으로부터 브라켓을 장착하고 있는 61개 치아에서 치균세균막을 채취하여 mutans streptococci를 MSB 배지에서 선택적으로 분리한 다음, 이들의 지놈 DNA를 추출하여 dextranase 유전자를 표적으로 하는 중합효소연쇄반응법을 시행하고, 그 증폭물을 Hae III로 소화하고, 이를 전기영동하여 제한 효소절편길이에 따라 그 종을 식별하였다. 또한 생물형을 조사하기 위하여 생화학적 검사를 실시하였다. 그 결과 브라켓과 치아 경계부 및 브라켓으로부터 2mm 이상 떨어진 평활면의 치면세균막에 존재하는 mutans streptococci 종은 서로 비슷한 검출 빈도를 보이나, 두 곳에 존재하는 mutans streptococci 생물형은 서로 차이가 있는 것으로 나타났다 향후 브라켓과 치아 경계부 및 치아 평활면의 치면세균막의 mutans Streptococci 생물형의 차이와 브라켓 주위의 법랑질 탈회 및 치아우식증 발병과의 상관관계에 대한 연구가 필요하다.

• 한국산학기술학회논문지
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• 제13권7호
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• pp.3267-3274
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• 2012

371 균주의 해양박테리아를 광양만에서 분리하였다. 우점종은 Pseudomonas aeruginosa, Aeromonas hydrophila, P. fluorescens, P. paucimobilis, Chryseomonas luteola, P. vescularis 등이었다. 영양염과 유기물을 제거할 수 있는 해양박테리아를 탐색하기 위하여 암모니아성 질소(100 mg/L), 질산성 질소(100 mg/L) 및 인(10 mg/L)이 각각 포함된 10 mL의 marine broth 2216 (DIFCO)에 해양박테리아를 접종(1.0%, v/v)하고 12시간 배양하였다. 스크리닝 테스트 결과 25% 이상 $COD_{Cr}$을 제거하는 해양박테리아는 16종, 15% 이상의 암모니아 질소를 제거하는 해양박테리아는 9종, 60% 이상의 질산 질소를 제거하는 해양박테리아는 11종 그리고 90% 이상의 인을 제거하는 해양박테리아는 13종이 분리되었다. Aeromonas hydrophila, Chryseomonas indologenes, Pseudomonas diminuta, Vibrio parahaemolyticus 균주가 유기물 및 영양염류 제거실험을 위해 선정되었다. 회분식 시험을 위해 4종의 해양박테리아를 $COD_{Cr}$ 250 mg/L, $NH_3-N$ 40 mg/L, ${NO_3}^{-}-N$ 40 mg/L, ${PO_4}^{3-}-P$ 10 mg/L이 각각 첨가되어 있는 변형 marine broth에 접종하고, 10시간 배양하면서 제거효율을 측정하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권10호
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• pp.1425-1429
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• 2005

Megaspahera elsdenii YJ-4, which was previously isolated as a producer of trans-10, cis-12 CLA, was studied for its carbon source on the CLA production. M. elsdenii YJ-4, was incubated with glucose and lactose, and cultured in batch and continuous culture systems with linoleic acid at various pHs to investigate CLA production. Batch cultures of the ruminal bacterium, M. elsdenii YJ-4, were resistant to stearic acid and linoleic acid, and little growth inhibition was observed even when the fatty acid concentration in the culture was as much as 4 mg $ml^{-1}$. Stationary phase batch cultures (0.25 mg bacterial protein $ml^{-1}$) that had been grown on lactate and incubated with linoleic acid (0.20 mg $ml^{-1}$) produced approximately 12 ${\mu}g$ trans-10, cis-12 CLA mg $protein^{-1}$ and little cis-9, trans-11 CLA was detected. Some linoleic acid was converted to hydrogenated products (chiefly stearic acid), but these fatty acids were less than 5 ${\mu}g$ mg bacterial $protein^{-1}$. Stationary phase batch cultures that had been grown on glucose produced at least 3-fold less trans-10, cis-12 CLA than ones grown on lactate. Cells from lactate-limited continuous cultures produced less trans-10, cis-12 CLA than those from batch culture, but only if the pH was greater than 6.4. When the pH of the lactate-limited continuous cultures was lower than 6.4, trans-10, cis-12 CLA and hydrogenated products declined. Cells from glucose-limited continuous cultures produced less trans-10, cis-12 CLA and hydrogenated products than the cells that had been limited by lactate, but pH had little impact on this production. These results support the idea that M. elsdenii YJ-4 could be one of the major producers of trans-10, cis-12 CLA which causes cows to produce milk with a low fat content.

• 대한치과교정학회지
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• 제24권4호
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• pp.871-883
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• 1994

This experiment was designed to study possible roles of $interleukin-1\beta$, interleukin-6 and tumor necrosis $factor-\alpha$ in bone remodeling by measuring their effects on $PGE_2,\; LTB_4$ and collagenase production when they were administered to human periodontal ligament fibroblasts. Human periodontal ligament fibroblasts were collected from first premolars extracted for orthodontic treatment. They were incubated in the environment of $37^{\circ}C,\;5\%\;Co^2,\;and\;100\%$ humidity. They were treated with $0.25\%$ trypsin-EDTA solution and centrifuged. PDL cells in the fifth to seventh passage were used for the experiment. Cells were seeded onto the culture dishes and when they were successfully attached, human recombinant $interleukin-1\beta$, interleukin-6, and tumor necrosis $factor-\alpha$ were administered, alone or in combination. They were incubated for 4, 8 and 24 hours and the levels of $PGE_2,\;LTB_4$ and collagenase released into the culture media were assessed by enzymeimmunoassay and collagenase activity assay. The conclusions are as follows: 1. $IL-1\beta\;and\;TNF-\alpha$ were very active in stimulating the production of $PGE_2$ and collagenase by human periodontal ligament fibroblasts, while IL-6 increased $LTB_4$ production. 2. $IL-1\beta$ significantly increased $PGE_2$, but $LTB_4$ Production was not increased. $IL-1\beta$ is thought to act mainly via the cyclooxygenase pathway of arachidonic acid metabolism. 3. IL-6 tended to inhibit $IL-1\beta$ in the production of $PGE_2$ and collagense whereas IL-6 and $TNF-\alpha$ showed auditive effect in the level of $PGE_2$. The above cytokines increased the release of at least one of $PGE_2,\;LTB_4$ and collagenase. It suggests that cytokines are involved in bone remodeling process by stimulating PDL fibroblasts to produce various bone-resorptive agents. The roles of cytokines in bone remodeling as a whole would need further study.

• Journal of Periodontal and Implant Science
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• 제26권4호
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• pp.873-887
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• 1996

Platelet-derived growth factor(PDGF) is one of the polypeptide growth fators. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. Recent studies indicated that demineralized root surface as the primary site for growth factor application has advantages over other application method, especially due to binding capacity of growth factor for exposed matrix component of deminera1ized dentin surface. The purpose of this study is to evaluate optimal application time of PDGF-BB on proliferation of human gingival fibroblasts using deminera1ized dentin surface as primary application site. Human gingival fibroblasts and dentin slabs were prepared from the first premolar tooth extracted for the orthodontic treatment, cells were cultured in DMEM/I0% FBS at the $37^{\circ}C$, 5% CO2 incubator. All of the dentin slabs were preconditioned with Tetracycline HCI(100mg/ml) solution and rinsed in PBS. In the cell proliferation experiment, experimental group was immersed in DMEM containing 10% FBS, 50ng/rnl PDGF-BB during different time(30sec, 1, 2, 4, 8 minutes) and dried. Cells at concentration of $1{\times}10^5$cells/ml were seeded in each culture well which contained dentin slabs and incubated for 6 hours. Then, all of the dentin slabs were moved into new 24 well culture dish and incubated for 24, 48, 72 hours. The cell counting was done by hemocytometer with inverted phase contrast microscope after trypsinization. The results were as follows : The application of PDGF-BB for 1, 2 min slightly increased the number of gingival fibroblasts, and the application of PDGF-BB for 4, 8 min prominently increased the number of gingival fibroblasts. The application of PDGF-BB for 4 min showed maximum proliferation rate of gingival fibroblasts at 24, 48, 72 hours, and the application of PDGF-BB for 8 min showed less proliferation rate of gingival fibroblasts compared to the application of PDGF-BB for 4 min at 24, 48, 72 hours. In conclusion, the application of PDGF-BB for 4 min appeared to be optimal to obtain maximum proliferation of gingival fibroblasts using demineralized dentin surface as primary applicaton site of PDGF-BB.

• Journal of Periodontal and Implant Science
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• 제37권1호
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• pp.115-124
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• 2007

Silica is known as a promising osteoconductive material, and polycaprolactone is a bioactive and degradable material. The purpose of this study was to monitor the differentiation of MC3T3-E1 cells cultured on the layer-built silica/poly caprolactone non-woven fabric produced by electrospinning. Non-woven fabric (silica, polycaprolactone, PSP, SPS) was made by electrospinning and they were inserted in the 48 well cell culture plate. MC3T3-E1 cells were prepared by subculture. Cells were seeded to each well $1{\times}10^5$ concentration per well. Dulbecco's modified eagle medium with 10% FBS and 1% antibiotic-antimycotic solution was used. Confocal laser scanning microscope was taken 4 hours after incubation (95% air. 5% $CO_2$, $37^{\circ}C$). Cell proliferation was monitored by spectrophotometer on 1, 7, 14 days, and the morphology of the growing cells was observed by field emission scanning electron microscope. To monitor the differentiation of osteoblasts on the materials, MC3T3-E1 cells were incubated in 48 well culture plate after seeding with the density of $1{\times}10^5$ concentration. Then ELISA kit & EIA kit were used on to assess osteocalcin and osteopontin expression respectively. The other conditions were the same as above. MC3T3-E1 cells were proliferated well on all of the materials. There were no statistical differences among them. The osteopontin expression of silica, PSP, SPS was significantly higher than other groups on day 3 (p/0,05), but after that time, there were no statistically signigicant differences. The osteocalcin expression was significantly higher in silica and PSP than other groups on day 14. These findings show that PSP was as good as silica on the effect of osteoblast differentiation. The PSP non-woven fabric may have the possibility as bone graft materials.

• 한국식품영양과학회지
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• 제36권12호
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• pp.1596-1603
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• 2007

연어 frame 추출물에 대하여 건강 기능성 개선을 위하여 4종의 상업적 효소(Alcalase, Flavourzyme, Neutrase 및 Protamex) 처리에 의한 건강 기능성 연어 frame 가수분해물의 개발을 시도하였다. 연어 frame 가수분해물의 ACE 저해능($IC_{50}$)은 Neutrase로 4시간 처리한 가수분해물이 0.67 mg/mL로 가장 우수하였으나, 항산화성은 기대 이하의 범위(15% 이하)이었다. Neutrase 처리 연어 frame 가수분해물은 무처리 추출물에 비하여 일반성분과 관능적 특성(비린내 및 맛)에 있어서 차이가 없었으나 추출물 질소 함량은 높았고, 투과도는 개선되었다. 효소처리 연어 frame 가수분해물은 축육과 뼈로 제조한 시판 곰탕에 비하여 단백질 함량, 추출물 질소, 구성아미노산과 칼슘 함량이 높았고, 투과도, 관능적 비린내 등에서는 차이가 없었으며, 추후 조미 등에 의하여 보강 가능한 관능적 맛은 기호도에 있어 약간 차이가 있었다. 이상의 결과로 미루어 보아 연어 frmae 추출물에 Neutrase로 4시간 처리함으로 인하여 투과도와 ACE 저해능과 같은 건강 기능성은 기대할 수 있으리라 판단된다.