• 한국약용작물학회지
• /
• 제18권3호
• /
• pp.157-167
• /
• 2010

We investigated a method to improve anticancer activities of Acer mono wood extracts by ultra high pressure extraction process. The A. mono was extracted by water at $40^{\circ}C$ and 300 MPa for 15 min (High Pressure Extraction, HPE). The extraction yield by ultra high pressure extraction process was 5.42%. The cytotoxicity on human normal lung cell (HEL299) of the extracts from HPE showed 21.54% lower than that from conventional water extraction at $100^{\circ}C$ in adding the maximum concentration of 1.0 mg/$m{\ell}$. Ultra high pressure extracts process for 15 minutes extracts (HPE15) showed more potent scavenging effect than the control, BHA. On SOD-like test, the HPE15 showed highest activity as 32.4% at 1.0 mg/$m{\ell}$ concentration. Human stomach adenocarcinoma, liver adenocarcinoma, breast adenocarcinoma and lung adenocarcinoma cell growth were inhibited up to about 67~79%, in adding 1.0 mg/$m{\ell}$ of extracts from HPE. HPE was 20~25% higher than conventional water extraction. It was interesting that, among several cancer cell lines (stomach adenocarcinoma, liver adenocarcinoma), the growth of digestive related cancer cells were most effectively inhibited as about 75~79%. On in vivo experiment using ICR mice, the variation of body weight of mice group treated A. mono wood extracts from HPE of 100 mg/kg/day concentration was very lower than control and other group. The survival times of group treated this extracts was 61.96% longer than that of the control group and this extracts showed the lower tumor weight, which were 10.49 g than positive control as 16.17 g. Based on these results, we could tell that the HPE wood extracts of A. mono had higher anticancer activity than conventional water extraction. The results of HPE showed obvious advantages in higher efficiency, shorter extraction time, at lower energy costs.

• 대한한의학회지
• /
• 제23권2호
• /
• pp.57-69
• /
• 2002

Objective: The aim of this study is to investigate the effect of Injin butanol fractions with Thin Layer Chromatography on Fas-mediated Apoptosis. Method: Injin-butanol fraction separated by TLC. MIT assay, cell cycle analysis, Caspase-3 protease assay, DNA fragmentation assay and quantitative RT-PCR were performed to evaluate the effects of TLC extraction of lnjin-butanol fraction on cell viability, cell cycle progression and apoptosis. Results: Scopoletin, luteolin, apigenin and unknown powder was isolated by TLC. Fas-mediated apoptosis analysis shows that scopoletin has inhibiting function on apoptosis. Caspase- 3 protease assay analysis shows that scopoletin inhibits activity of caspase-3. Quantitative RT-PCR analysis shows that no activity on caspase-3, but apoptosis inhibition cytokine -Bcl-2- is activated, and apoptosis activating cytokine -Bax- is unactivated. Conclusion: These results show that each fraction of Injin-butanol TLC extraction, especially scopoletin, acts as a protective function on liver cell viability, and inhibitory function on apoptosis. (J Korean Oriental Moo 2002;23(2):57-69)

• Korean Chemical Engineering Research
• /
• 제54권1호
• /
• pp.135-139
• /
• 2016

본 연구에서는 무기염을 첨가한 액-액 추출에 의해 식물세포인 바이오매스로부터 파클리탁셀 회수 방법을 획기적으로 개선하고자 하였다. 다양한 무기염(NaCl, KCl, $K_2HPO_4$, $NaH_2PO_4$, $NaH_2PO_4{\cdot}2H_2O$)을 이용하여 추출효율을 조사한 결과, NaCl에서 가장 낮은 분배계수(0.053)로 가장 높은 파클리탁셀 수율(~96%)을 얻을 수 있었다. NaCl을 이용한 액-액 추출에서 최적의 NaCl/용매 비와 메틸렌 클로라이드/메탄올 비는 각각 1%(w/v)와 26%(v/v)이었다. 또한 최적의 NaCl/용매 비와 메틸렌 클로라이드/메탄올 비에서 파클리탁셀 함량에 따른 영향을 조사한 결과, 순수 파클리탁셀 함량 0.066%(w/v)에서 가장 낮은 분배계수(0.053)로 가장 높은 수율(~96%)을 얻을 수 있었다. 기존 액-액 추출의 경우 총 3회의 추출로 파클리탁셀을 95% 정도 회수 가능한 반면 무기염을 이용한 방법의 경우 단 1회 추출로 대부분의 파클리탁셀을 회수(~96%) 가능하였다.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2002년도 생물공학의 동향 (X)
• /
• pp.112-115
• /
• 2002

본 연구에서는 균주의 배양과 동시에 생산물을 회수할 수 있는 동시추출공정을 건조질량의 $15{\sim}75$ %의 탄화수소를 생산한다고 알려진 B. braunii 배양에 적용하고자 한다. 일반적인 tow-phase 동시 추출공정의 적용시 B. braunii의 경우 생산된 탄화수소가 균주 외벽의 matrix에 강하게 부착되어 있기 때문에, two phase 추출공정 적용시 bubble solumn내에서 단지 폭기에 의한 교반만으로는 충분한 탄화수소의 회수율을 얻을 수가 없었다. 본 연구에서는 배양액과 유기용매층의 접촉기회를 증대시킨 two-stage 동시추출 공정을 개발하여 기존의 two-stage 동시추출 공정보다 2배 이상 높은 57 %의 탄화수소 회수율을 얻을 수가 있었고, 이를 회분배양후 후속분리공정으로 이용할 경우 6시간 추출후 62 %의 회수율을 얻을 수가있었다.

• 대한약침학회지
• /
• 제14권1호
• /
• pp.79-86
• /
• 2011

Objective : This study was performed to compare anti-inflammation, anti-oxidation and anti-bacterial effects of Cuscutae Semen(CS) extracted with two kinds of solvents, ethanol and distilled water. Methods : Two kinds of CS extractions were prepared 20, 50, 100 ${\mu}l/mg$. The cytotoxicity was measured by MTT assay in Raw 264.7 cell. The anti-inflammation effect was measured by inhibitory efficacy of NO Production in Raw 264.7 cell. The anti-oxidation effect was measured by DPPH Radical scavenging ability in HaCaT cell. The anti-bacterial effect was measured by inhibition zone diameter on Propionibacterium acnes. Results : 1. Two kinds(100 ${\mu}l/mg$) of CS extraction groups had 50% cytotoxicity in Raw 264.7 cell. 2. All of CS extraction groups were not showed significantly inhibitory effect on NO production. 3. All of CS extracted with ethanol only showed dose-dependently significantly scavenging effect of DPPH radicals. 4. Two kinds of CS extractions did not have a inhibitory effect on Propionibactrium acnes. Conclusion : Two kinds(100 ${\mu}l/mg$) of CS extraction groups have 50% cytotoxicity. Two kinds of CS extractions have not the inhibitory effect on NO production and Propionibactrium acnes. CS groups extracted with ethanol only have a significantly scavenging ability of DPPH radicals. This study suggests that CS extracted with ethanol was effective in anti-oxidation.

• 한국미생물·생명공학회지
• /
• 제48권2호
• /
• pp.129-137
• /
• 2020

This study aims to evaluate the efficacy of the Ultrasonication-Assisted (UA) extraction on the functionality of the herbaceous biennial plant maca (Lepidium meyenii). The specific objectives include comparison of the antioxidant activities among various maca extracts, determination of the macamide B content of the extracts, and in vitro evaluation of maca on cell viability and creatine kinase (CK) activity. The antioxidant activities of the water, ethanol, and UA extracts were compared by determining the total phenolic and flavonoid contents, the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities, and the ferric reducing antioxidant power (FRAP) of the extracts. The macamide B content of maca extracts were analyzed by HPLC. The effects of the extracts on muscle cell viability and creatine kinase activity were also determined using C2C12 myoblasts. UA extraction significantly increased the total phenolic content (2.90 GAE ㎍/mg, p < 0.05), without affecting the flavonoid content. DPPH radical scavenging activity did not exhibit any statistical difference among the extracts. The ethanol and UA extracts exhibited significantly higher FRAP than the water extract (p < 0.05). The macamide B content of ethanol and UA extracts were 0.087 and 0.083 ㎍/mg, respectively. The water and UA extracts exhibited higher C2C12 muscle cell viability than the ethanol extract, and both extracts resulted in a significantly lower CK level than the H₂O₂-treated control group. This research suggests that the maca extract can protect muscle cells and serve as an antifatigue agent under oxidative stress conditions.

• 한국약용작물학회지
• /
• 제16권6호
• /
• pp.383-389
• /
• 2008

This study was performed to compare effect of immune activities of Rhodiola sachalinensis by various extraction process with different temperature and extraction solvents. Experiments were performed for investigate the immune activities on human B and T cell growth and secretion of their cytokines. Also, antibodies in serum were investigated in female ICR mouse by feeding the extracts of R. sachalinensis at doses of 40, 120 and 360 mg/kg orally for 15 days. The immune cell growth and secretion of cytokines (IL-6, TNF-$\alpha$) on human B and T cells were increased by adding R. sachalinensis extracts compare to the control. Also, total serum IgG levels increased by feeding R. sachalinensis extracts. It can be conclude that optimum condition for efficient extraction of R. sachalinensis as functional material is slovent extraction process using water with ultrasonification at below $100^{\circ}C$ than typical process.

• Asian-Australasian Journal of Animal Sciences
• /
• 제30권11호
• /
• pp.1620-1632
• /
• 2017

Objective: This study evaluated the effects of a traditional Chinese medicine formula (TCMF) on muscle fiber characteristics in finishing pigs and the effects of the formula's extract (distilled water, ethyl acetate and petroleum ether extraction) on porcine cell proliferation and isoforms of myosin heavy chain (MyHC) gene expression in myocytes. Methods: In a completely randomized design, ninety pigs were assigned to three diets with five replications per treatment and six pigs per pen. The diets included the basal diet (control group), TCMF1 (basal diet+2.5 g/kg TCMF) and TCMF2 (basal diet+5 g/kg TCMF). The psoas major muscle was obtained from pigs at the end of the experiment. Muscle fiber characteristics in the psoas major muscle were analyzed using myosin ATPase staining. Cell proliferation was measured using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) dye and cytometry. Isoforms of MyHC gene expression were detected by real-time quantitative polymerase chain reaction. Results: The final body weight and carcass weight of finishing pigs were increased by TCMF1 (p<0.05), while the psoas major muscle cross-sectional area was increased by TCMF (p<0.05). The cross-sectional area and diameter of psoas major muscle fiber Ι, IIA, and IIB were increased by TCMF2 (p<0.05). The cross-sectional area and fiber diameter of psoas major muscle fiber IIA and IIB were increased by diet supplementation with TCMF1 (p<0.05). Psoas major muscle fiber IIA and IIB fiber density from the pigs fed the TCMF1 diet and the type IIB fiber density from the pigs fed the TCMF2 diet were lower compared to pigs fed the control diet (p<0.05). Pigs fed TCMF2 had a higher composition of type Ι fiber and a lower percentage of type IIB fiber in the psoas major muscle (p<0.05). The expression levels of MyHC Ι, MyHC IIa, and MyHC IIx mRNA increased and the amount of MyHC IIb mRNA decreased in the psoas major muscle from TCMF2, whereas MyHC Ι and MyHC IIx mRNA increased in the psoas major muscle from TCMF1 (p<0.05). Peroxisome proliferator-activated receptor ${\gamma}$ $coactivator-1{\alpha}$ and CaN mRNA expression in the psoas major muscle were up-regulated by TCMF (p<0.05). Porcine skeletal muscle satellite cell proliferation was promoted by $4{\mu}g/mL$ and $20{\mu}g/mL$ TCMF water extraction (p<0.05). Both $1{\mu}g/mL$ and $5{\mu}g/mL$ of TCMF water extraction increased MyHC IIa, MyHC IIb, and MyHC IIx mRNA expression in porcine myocytes (p<0.05), while MyHC Ι mRNA expression in porcine myocytes was decreased by $5{\mu}g/mL$ TCMF water extraction (p<0.05). Porcine myocyte MyHC Ι and MyHC IIx mRNA expression were increased, and MyHC IIa and MyHC IIb mRNA expression were down-regulated by $5{\mu}g/mL$ TCMF ethyl acetate extraction (p<0.05). MyHC Ι and MyHC IIa mRNA expression in porcine myocytes were increased, and the MyHC IIb mRNA expression was decreased by $1{\mu}g/mL$ TCMF ethyl acetate extraction (p<0.05). Four isoforms of MyHC mRNA expression in porcine myocytes were reduced by $5{\mu}g/mL$ TCMF petroleum ether extraction (p<0.05). MyHC IIa mRNA expression in porcine myocytes increased and MyHC IIb mRNA expression decreased by $1{\mu}g/mL$ in a TCMF petroleum ether extraction (p<0.05). Conclusion: These results indicated that TCMF amplified the psoas major muscle cross-sectional area through changing muscle fiber characteristics in finishing pigs. This effect was confirmed as TCMF extraction promoted porcine cell proliferation and affected isoforms of MyHC gene expression in myocytes.

• Animal cells and systems
• /
• 제13권3호
• /
• pp.351-356
• /
• 2009

Ciliates are undoubtedly one of the most diverse protozoans that play a significant role in ecology. However, molecular examination, based on comparing the DNA sequences, has been done on a limited number of the species. Because most ciliates are uncultivable and their population sizes are often too small, it is usually difficult to obtain sufficient genomic DNA required for PCR based experiments. In the present study, we evaluated the effectiveness of four commercial DNA extraction procedures that extract high quality genomic DNA from a single ciliate cell. It was discovered that RED Extract-N-$Amp^{TM}$ PCR kit is the best method for removing PCR-inhibiting substances and minimizing DNA loss during purification. This method can also amplify more than 25 reactions of PCR. In addition, this technique was applied to single cells of 19 species belonged to 7 orders under 5 classes that isolated from mixed natural populations. Their small subunit ribosomal DNA (SSU rDNA) was successfully amplified. In summary, we developed a simple technique for the high-yield extraction of purified DNA from a single ciliate cell that may be more useful for rare ciliates, such as tiny and uncultivable marine microbes.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권6호
• /
• pp.2733-2737
• /
• 2014

Objective: Molecular pathology tests are often carried for clinicopathological diagnosis and pathologists have established large collections of formalin-fixed, paraffin-embedded tissue (FFPE) banks. However, extraction of DNA from FFPE is a laborious and challenging for researchers in clinical laboratories. The aim of this study was to compare two widely used DNA extraction methods: using a QIAamp DNA FFPE kit from Qiagen and a Cobas Sample Preparation Kit from Roche, and evaluated the effect of the DNA quality on molecular diagnostics. Methods: DNA from FFPE non-small cell lung carcinoma tissues including biopsy and surgical specimens was extracted with both QIAamp DNA FFPE and Cobas Sample Preparation Kits and EGFR mutations of non-small cell lung carcinomas were detected by real-time quantitative PCR using the extracted DNA. Results and Conclusion: Our results showed that DNA extracted by QIAamp and Cobas methods were both suitable to detect downstream EGFR mutation in surgical specimens. Howover, Cobas method could yield more DNA from biopsy specimens, and gain much better EGFR mutation results.