• 제목/요약/키워드: freeze-dried cells

검색결과 140건 처리시간 0.028초

오만둥이(Styela plicata) 추출물의 in vitro 세포독성 효과 (In vitro Cytotoxic Effect of Extracts from Styela plicata)

  • 이보배;차미란;박해룡;이승철
    • 한국식품영양과학회지
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    • 제36권9호
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    • pp.1099-1105
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    • 2007
  • 오만둥이의 세포독성 효과를 알아보기 위해 신선한 오만둥이를 분쇄하고 동결건조하여 분말 형태로 제조한 후 시료 5 g당 메탄올, 에탄올, 아세톤, 물을 각각 100 mL씩 첨가하여 용매별 추출물을 제조하였다. 이 용매별 추출물을 대장암 세포주인 HT-29 cell에 250 ${\mu}g/mL$ 농도로 각각 처리하여 형태학적인 변화를 관찰한 결과 대조군에 비해 아세톤 추출물에서 세포의 수가 감소하고 그 형태가 응축되는 현상을 관찰할 수 있었고, 이 결과를 바탕으로 100, 250, 500 ${\mu}g/mL$ 농도로 MTT reduction assay를 실시한 결과 아세톤 추출물을 500 ${\mu}g/mL$ 농도로 처리했을 때 6%의 암세포만이 생존하는 것으로 나타나 형태학적인 실험결과와 동일한 결과를 얻을 수 있었다. 아세톤 추출물을 헥산, 디에틸 에테르, 에틸 아세테이트 그리고 물로 분획물을 조제하여 대장암 세포주인 HT-29 cell에 250 ${\mu}g/mL$ 농도로 각각 처리하여 형태학적인 변화를 관찰한 결과 헥산 분획물에서 세포의 형태변화가 가장 크게 나타났으며 250 ${\mu}g/mL$ 농도로 MTT reduction assay를 실시한 결과에서도 5.1%의 암세포만이 존재하는 것으로 나타나 헥산 분획물이 가장 높은 세포독성 효과를 보이는 것을 확인할 수 있었다. 다른 암세포주에 대한 영향을 알아보기 위해 헥산 분획물을 SW620, HeLa 및 MCF-7 세포주에 농도별로 처리한 결과, 250 ${\mu}g/mL$ 농도에서 10% 내외의 암세포 생존율을 보여 높은 세포독성 효과를 나타내었으며 특히, HeLa 세포주에서는 생존율이 5.8%로 나타나 매우 높은 활성을 보였다. 이상의 결과를 종합하여 미루어 짐작해 볼 때 오만둥이에 함유된 불포화 지방산 등의 지용성 물질이 암세포주의 성장 억제에 중요한 역할을 하리라 생각된다. 따라서 오만둥이 추출물은 다양한 암세포주에 대해 활성을 가지는 해양생물로서의 잠재적인 가능성을 가진다는 것을 확인할 수 있었다.

악골결손 재건을 위한 탈회 및 비탈회 동결건조 동종골의 이용 (USE OF DEMINERALIZED AND MINERALIZED FREEZE-DRIED ALLOGENIC BONE GRAFT FOR THE CORRECTION OF MAXILLOFACIAL DEFORMITIES; CASE REPORTS)

  • 이기혁;여환호;김영균;김수관;이병준;박인순;엄인웅
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제18권3호
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    • pp.371-377
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    • 1996
  • Bone graft has been used to repair one defect caused by disease and trauma, congenital and acquired deformities. Graft materials are autogenous bone, allogenic bone, xenogenic bone, synthetics. Autogenous bone graft is the most superior to other materials for immunologic reaction, compatibility to host tissue, and revascularization. However, autogenous bone graft is required for additional operation and the amount of taking is limited. Autografts are obtained at own expense and also limited in size, shape. In order to compensate these problems, allogenic bone graft has been used increasingly. But allogenic bone graft encounters immunologic complications. Therefore, it has been used after freezing, lyophilization, or demineralization. Allogenic bone processed by only lyophilization includes potential antigenic properties on its surface, therefore it is demineralized to deplete immunologic reaction. Demineralized bone releases BMP and helps the mesenchymal cells transform to the chondroblast to produce cartilage and bone. This reaction is called osteoinducation. Many authors have reported that mineralized lyophilized bone had less antigenicity clinically and favorable bony consideration with host bone. In our department from 1995 to now, we have used banked allogenic bone graft that has been prepared from Wonkwang Bone Bank in 5 cases and mineralized lyophilized bone graft in 2 cases to reconstruct the maxillofacial bone defect after tumor resection and cyst enucleation and cleft alveolus. We will report with literature review that the result is favorable functionally and esthetically.

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Lactobacillus sp.균주를 이용한 산삼 배양근 발효물의 기능성 평가 (Evaluation of Functional Properties of the Tissue Cultured Wild Ginseng Fermented by Lactobacillus sp.)

  • 신은지;조장원;김영언;한대석;홍희도;이영경
    • 한국식생활문화학회지
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    • 제27권6호
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    • pp.743-750
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    • 2012
  • A tissue cultured wild ginseng (TCWG) suspension was inoculated with lactic acid bacteria and fermented to improve the functionality of TCWG. The utilization of TCWG was increased directly using the freeze-dried powder. The optimal ratio of TCWG powder and water for fermentation was 1:19 (5%), which was selected by measuring the fluidity and viable cell count according to concentration. The effects on ADH activation and immune cell activation by each ferments with 10 kinds of Lactobacillus sp. strains were examined. The ferments with the Lactobacillus casei KFRI 692 strain showed 5.4 times higher ADH activity and 1.3 times higher ALDH activity than the non-fermented TCWG powder (control). The level of NO production and cytotoxicity was also measured by Raw 264.7 cells. The ferment with the Lac. casei KFRI 692 strain showed the highest level of NO production and lower cytotoxicity than the others. Therefore, the Lac. casei KFRI 692 strain was selected as a strain for fermentation of a TCWG suspension to maximize its functionality. To identify the optimal fermentation time of the selected Lac. casei KFRI 692 strain on the 5% TCWG suspension, the viable cell count of lactic acid bacterial and the changes in pH were observed for 72 hours. 24-hrs was found to be the optimal fermentation time. In this way, fermented TCWG with lactic acid bacteria showed higher ADH activation efficacy and immune cell activation than non-fermented TCWG.

천년초선인장의 부위별 영양성분 분석과 정상 및 암세포에 미치는 효과 (Nutrients Contents in Different Parts of Pickly Pear(Opuntia humifusa) and Possible Anti-Breast Cancer Effect)

  • 윤진아;함상욱;손용석
    • 한국식품영양학회지
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    • 제22권4호
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    • pp.485-491
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    • 2009
  • In order to examine the nutritional value of prickly pear(Opuntia humifusa), contents of ash, protein, fat, minerals and vitamins were determined on freeze-dried stem, fruit, seed and root from plants harvested in autumn. The average moisture contents for stem, fruit, seed and root were 67~87%. Crude ash content determined on dry weight basis was 2~3%. Crude protein existed mostly in seed(2.95%) and root(2.37%). Crude fat was detected mainly in seed(4.49%). Contents of major minerals(mg/100 mg dry weight) was generally higher in stem. Ca in stem(4,142.30) and fruit(2,790.86) were much higher than in seed(43.37). P in stem, seed and fruit were 448.19, 263.20 and 161.59, respectively. Stem also displayed more abundant Mg(1,110.86), Zn(35.62) and Mn(37.07). However, fruit contained higher amounts of Fe(13.38) and Se(0.15). Vitamin A was negligible in all plant parts. Vitamin E contents in fruit and stem were 1.78 mg and 1.22 mg/mg dry weight, respectively. Vitamin C was detected mostly in fruit(445.40) and stem(260.94). Use of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-based microtiter assay of cell viability demonstrated an anti-proliferative effect of O. humifusa extract on the MCF-7 estrogen-dependent human breast cancer cell line.

천연물중의 생리활성성분이 간해독기구에 미치는 영향 (Effects of Biologically Active Substances in Natural Products on the Hepatic Detoxication Mechanism)

  • 권정숙
    • Journal of Nutrition and Health
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    • 제27권4호
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    • pp.347-355
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    • 1994
  • Indolo[3,2-b]carbazole(ICZ) is a potent Ah receptor agonist with biological activities similar in several respects to those of the potent environmental toxin, TCDD. ICZ is produced during the oilgomerization of indole-3-carbinol(I3C), a breakdown product of the glucobrassicin present in food plants of the Brassica genus. In the present study we examined ICZ levels in tissues and excreta of rats treated with I3C or dietary cabbage of established glucobrasicin content, and in feces of conventional and germfree rats fed on a basal diet, and of humans. We also examined the levels of cytochrome P4501A1 induction, as determined by the ethoxyresorufin ο-deethylase assay, in tissues of animals that received cabbage-supplemented diets, or which were treated with purified I3C or ICZ. Our findings indicated that incorporation of either homogenized or whole freeze-dried cabbage in the feed led to large increases(16-60 fold) in the levels of ICZ in the feces and lower gastrointestinal tract of rats. We observed that whereas ICZ is readily detectable at about the same levels(2.00$\pm$0.50 ppb) in the feces of conventional rats fed on a purified diet and in human feces, levels of ICZ in the feces of germfree animals fed on the basal diet were at the limits of detection(0.40$\pm$0.20 ppb), indication that gut bacteria are important for the production of ICZ from essential dietary constituents in the basal diet. We showed that in contrast to the near 7000-fold difference in CYP1A1 inducing potencies of ICZ and TCDD in cells in culture, their inducing potencies differ by only about an order of magnitude in rats. Nonetheless, the levels of ICZ remaining in livers twenty hours after I3C treatment appear too low to account for the induced activity. This result indicates that ICZ may be rapidly cleared from the liver or that substances other than, or in addition to, ICZ be responsible for the enzyme-inducing activity of orally administered I3C or its precursors.

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Anti-inflammatory activity of Ganoderma lucidum by inhibition of NF-κB p65 phosphorylation

  • Kim, Hyung Don;Park, Jeong-Yong;Noh, Hyung-Jun;Lee, Seung Eun;Lee, Jeong Hoon;Seo, Kyung Hye
    • 농업과학연구
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    • 제46권3호
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    • pp.653-660
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    • 2019
  • Ganoderma lucidum, an oriental polypore fungus and medicinal mushroom, has a long history of use for promoting health and longevity in Korea, China, and other Asian countries. This study was aimed at determining the anti-inflammatory activity and mechanism of action of Ganoderma lucidum in murine macrophage RAW 264.7 cells. Ganoderma lucidum was extracted with ethanol and freeze-dried. The anti-inflammatory effect (nitrite production) of Ganoderma lucidum extracts was tested using a nitric oxide (NO) colorimetric assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression of cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6. Western blotting was performed to measure the expression levels of inflammation-related proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor kappa B ($NF-{\kappa}B$) p65, and phosphorylated $NF-{\kappa}B$ p65. The NO colorimetric assay showed that NO production increased with the treatment of lipopolysaccharide in (LPS)-activated RAW 264.7 macrophages and decreased with the cotreatment of Ganoderma lucidum extracts and LPS. Ganoderma lucidum extracts repressed the mRNA expressions of cytokines, which were increased after the LPS treatment. In addition, Ganoderma lucidum extracts inhibited the LPS-induced expression of iNOS and COX-2 and the LPS-induced phosphorylation of $NF-{\kappa}B$ p65. These results suggest that the Ganoderma lucidum extracts exert an anti-inflammatory activity by inhibiting $NF-{\kappa}B$ related proteins and cytokines.

Determination of Optimized Growth Medium and Cryoprotective Additives to Enhance the Growth and Survival of Lactobacillus salivarius

  • Yeo, Soyoung;Shin, Hee Sung;Lee, Hye Won;Hong, Doseon;Park, Hyunjoon;Holzapfel, Wilhelm;Kim, Eun Bae;Huh, Chul Sung
    • Journal of Microbiology and Biotechnology
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    • 제28권5호
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    • pp.718-731
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    • 2018
  • The beneficial effects of lactic acid bacteria (LAB) have been intensively investigated in recent decades with special focus on modulation of the host intestinal microbiota. Numerous discoveries of effective probiotics are driven by a significantly increasing demand for dietary supplements. Consequently, technological advances in the large-scale production and lyophilization are needed by probiotic-related industries for producing probiotic LAB for commercial use. Our study had a dual objective, to determine the optimum growth medium composition and to investigate appropriate cryoprotective additives (CPAs) for Lactobacillus salivarius, and compare its responses with other Lactobacillus species. The one-factor-at-a-time method and central composite design were applied to determine the optimal medium composition for L. salivarius cultivation. The following composition of the medium was established (per liter): 21.64 g maltose, 85 g yeast extract, 1.21 ml Tween 80, 6 g sodium acetate, $0.2g\;MgSO_4{\cdot}7H_2O$, $0.02g\;MnSO_4{\cdot}H_2O$, $1g\;K_2HPO_4$, $1.5g\;KH_2PO_4$, $0.01g\;FeSO_4{\cdot}7H_2O$, and 1 g sodium citrate. A cryoprotective additive combination comprising 10% (w/v) skim milk and 10% (w/v) sucrose supplemented with 2.5% (w/v) sodium glutamate was selected for L. salivarius, and its effectiveness was confirmed using culture-independent methods in the freeze-dried cells of the Lactobacillus strains. In conclusion, the optimized medium enhanced the species-specific cultivation of L. salivarius. On the other hand, the cryoprotective effects of the selected CPA mixture may also be dependent on the bacterial strain. This study highlights the necessity for precise and advanced processing techniques for large-scale production of probiotics in the food and feed industries.

혈관내피세포에서 고농도 포도당으로 유도된 산화스트레스에 대한 조릿대잎 추출물의 보호효과 (Protective Effects of Sasa Borealis Leaves Extract on High Glucose-Induced Oxidative Stress in Human Umbilical Vein Endothelial Cells)

  • 황지영;한지숙
    • 한국식품영양과학회지
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    • 제39권12호
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    • pp.1753-1760
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    • 2010
  • 혈관내피세포는 만성적인 고혈당 상태에 노출되면, 반응성 산소기의 급격한 증가로 인해 산화스트레스를 일으키게 된다. 이를 보호하기 위해 사용되는 항산화제는 혈관내피세포의 생존율을 높이며 2차적인 질병의 유발을 감소 및 완화시켜서 당뇨합병증에 도움이 되는 것으로 알려져 있는데, 본 연구에서는 항산화 효과가 있는 것으로 알려진 조릿대잎 추출물을 이용하여 고농도의 포도당으로 유도된 산화스트레스에 대한 혈관내피세포의 보호효과를 조사하고자 하였다. 즉, 30 mM 포도당에 노출되어 산화적 스트레스가 유발된 HUVECs에 조릿대잎 추출물 ethyl acetate(ESLE) 층을 분주한 결과, 농도 의존적으로 세포 생존율이 증가하였고, 활성산소종 수준과 지질과산화물 가는 ESLE 첨가에 의해 유의적으로 감소하는 것을 확인할 수 있었다. 또한 고농도 포도당으로 GSH 함량과 SOD, GSH-px, catalase 등과 같은 항산화효소의 활성이 감소된 HUVECs에 ESLE를 분주하였을 때 이들의 함량 및 활성이 유의적으로 증가되었으며, western blotting을 통해 ESLE 첨가가 SOD와 catalase 발현을 증가시킴을 확인할 수 있었다. 따라서 ESLE는 혈관내피세포에서 고농도 포도당으로 인해 유발된 산화적 스트레스에 대해 세포를 보호하는 효과가 있는 것으로 나타났으며, 이는 ESLE가 세포내 활성산소종을 감소시키고 항산화효소계의 활성을 증가시킴으로써 산화스트레스를 감소시킨 결과에 기인하는 것으로 사료되었다.

ICR 마우스 모델을 이용한 녹용 추출물의 생화학적 평가 및 급성 경구 독성을 포함한 세포 독성 효과 (Biochemical Assessment of Deer Velvet Antler Extract and its Cytotoxic Effect including Acute Oral Toxicity using an ICR Mice Model)

  • 칠라칼라 라마크리시나;문현정;이환;이동성;정선희
    • 한국식품위생안전성학회지
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    • 제38권6호
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    • pp.430-441
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    • 2023
  • 녹용은 수많은 연구에서 면역력 강화를 포함한 영양 및 의학적 가치를 입증하였으며 전통적인 약으로 널리 사용되고 있다. 본 연구는 녹용 추출물 (sample 1: 생녹용 추출물, sample 2: 건녹용 추출물, sample 3: 동결 건조 추추물)의 일반성분과 우론산, 황산화 글리코사미노글리칸, 시알릭산, 콜라겐을 포함한 유효성분을 조사하고, 액체 크로마토그래피quadrupole-time-of-light mass spectrometry (UPLC/QTOFMS)를 사용하여 녹용 추출물의 화학 성분을 분석하는데 목적이 있다. 또한, HT22 해마 세포, BV2 미세아교세포, RAW264.7 대식세포 및 HaCaT 케라틴 세포를 사용하여 MTT 분석을 통해 녹용 추출물의 세포 독성 효과 평가와 암컷과 수컷 ICR 마우스에 녹용 추출물을 각각 (0, 500, 1000, 2000 mg/kg) 경구투여 하여 급성 독성평가를 실시하였다. 투여후에는 OECD 가이드라인에 따라 마우스의 일반독성, 생존율, 체중 변화, 사망률, 임상 징후 및 부검 결과를 관찰하였다. 결과적으로 녹용 추출물은 HaCaT 케라틴 세포에서 세포 독성 효과가 없었으며, 건녹용 추출물에서는 HT22 해마 세포에서 500 ㎍/mL, RAW264.7 대식세포의 경우 1000 ㎍/mL 에서, 동결건조추출물에서는 RAW264.7 세포와 BV2 미세아교세포의 경우 500 ㎍/mL 및 1000 ㎍/mL 농도에서 세포 독성을 가지고 있음을 보였다. 그러나 마우스를 이용한 급성 독성 평가에서는 녹용 추출물 시료를 처리한 모든 마우스에서 사망률, 임상 징후 및 부검 결과 특이사항이 없었으며 이는 LD50이 2000 mg/kg 이상으로 사료된다. 그러나 인간에 대한 안전성에 대한 충분한 증거를 확보하기 위해서는 동물과 사람에 대한 추가적인 연구가 필요하다.

Kefir 배양용 기능성 복합 Starter 개발 (Development of a Functional Mixed-Starter Culture for Kefir Fermentation)

  • 이봄이;이해창;문용일;오세종
    • Journal of Dairy Science and Biotechnology
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    • 제36권3호
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    • pp.178-185
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    • 2018
  • Kefir, which originates in the Caucasian mountains, is a cultured milk beverage produced by a combination of acidic and alcoholic fermentation. Kefir products are commonly used as food vehicles to deliver health-promoting materials including kefran and lactic acid bacteria to consumers. The aim of this study was to develop a freeze-dried starter culture without yeast and assess the suitability of kefir-like dairy products for the growth of lactic acid bacteria and the acidification of milk. Pasteurized whole milk (SNF 8.5%) stored at $25^{\circ}C$ was aseptically inoculated with starter cultures (0.002% w/v); it was kept at $25^{\circ}C$ until the pH attained a value of 4.6. Ten grams of the kefir-like product sample was diluted with 90 mL of 0.15% peptone water diluent in a milk dilution bottle, followed by uniform mixing for 1 min. Viable cells of Lactobacillus species were enumerated on modified-MRS agar (pH 5.2), with incubation at $37^{\circ}C$ for 48 h. Viable cells of Lactococcus species were enumerated on M17-lactose agar, with incubation at $32^{\circ}C$ for 48 h. The pH attained a value of 4.6 after fermentation for 9 h 30 min (Starter 1), 9 h 45 min (Starter 2), and 12 h (Starter 3). The viable cell count of Lactobacillus sp. and Lactococcus sp. was initially $10^5{\sim}10^6CFU/g$; it increased significantly to $10^9CFU/g$ after 12 h of incubation. During the storage of the kefir-like products at $4^{\circ}C$ for 1 4 days, the total viable cell numbers were unchanged, but the pH decreased slightly. The consistency of the kefir products increased gradually during the storage. The organoleptic properties of the kefir products fermented using the new starter culture are more desirable than those of commercial kefir. These results suggest that the newly developed starter culture without yeast could be suitable for kefir fermentation.