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Chemical Composition and Antitumor Apoptogenic Activity of Methylene Chloride Extracts from the Leaves of Zanthoxylum schinifolium (Zanthoxylum schinifolium잎의 methylene chloride 추출물의 화학적 조성 및 암세포에 대한 세포자살 유도활성과 그 작용기전)

  • Kim Jun-Seok;Jun Do-Youn;Woo Mi-Hee;Rhee In-Koo;Kim Young-Ho
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.546-554
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    • 2006
  • To understand antitumor activity of Zanthoxylum schinfolium, which has been used as an aromatic and medicinal plant in Korea, the cytotoxic effect of various organic solvent extracts of its leaves on human tumor cells were investigated. Among these extracts such as methanol extract (SL-13), methylene chloride extract (SL-14), ethyl acetate extract (SL-15), n-butanol extract (SL-16), and residual fraction (SL-17), SL-14 appeared to contain the most cytotoxic activity against leukemia and breast cancer cells tested. The methylene chloride extra.1 (SL-14) possessed an apoptogenic activity causing apoptotic DNA fragmentation of human acute leukemia Jurkat T cells via mitochondrial cytochrome c release into cytoplasm, subsequent activation of caspase-9 and caspase-3, and cleavage of PARP, which could be negatively regulated by antiapoptotic protein Bcl-xL. The GC-MS analysis of SL-14 revealed that the twenty-two ingredients of SL-14 were 9,19-cyclolanost-24-en-3-ol (15.1%), 2-a-methyl-17, b-hop-21-ene (15.1%), 15-methyl-2,3-dihydro-1H benzazepin (11.95%), phytol (10.38%), lupeol (9.92%), 12-methylbenzofuran (8.23%), hexadecanoic acid (5.96%), cis,cis,cis-9,12,15-octadecatrienoic acid-methyl-ester (5.49%), 9,12,15-octadecatrienoic acid-methylester (3.59%), 15-methyl-4-(1-methylethylidene)-2-(4-nitrophenyl) (3.36%), hexadecanoic acid methyl ester (1.93%), vitamine E (1.88%), beta-amyrin (0.96%), and auraptene (0.89%). These results demonstrate that the cytotoxicity of the methylene chloride extract of the leaves of Z. schinifolium toward Jurkat T cells is mainly attributable to apoptosis mediated by mitochondria-dependent caspase cascade regulated by Bcl-xL, and provide an insight into the mechanism underlying antitumor activity of the edible plant Z. schinifolium.

Identification of Maysin and Related Flavonid Analogues in Corn Silks (옥수수 수염에서 Maysin 및 유사물질의 동정)

  • Kim, Sun-Lim;Snook, Maurice E.;Kim, E-Hun;Park, Cheol-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.3
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    • pp.151-157
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    • 2000
  • This study was carried out to isolate and identify the maysin and related flavonoid analogues in corn silks. Silks were covered with silk bag to prevent pollination and were sampled at 3-5 days after silking. The silks were filled with 100% MeOH and stored at $0^{\circ}C$ until analysis. The MeOH extracts of corn silks were filtered and concentrated at 35-4$0^{\circ}C$. The ${CH}_2$${Cl}_2$ was added on the concentrated aqueous solution to remove the chlorophyll and lipids. The Cis open column (25mm$\times$54 cm) was washed and activated with serial treatment of 500$m\ell$ of 100% MeOH(twice)longrightarrow75% MeOH longrightarrow50% MeOHlongrightarrow30% MeOHlongrightarrow100% $H_2$O(2 times). The concentrated aqueous solution was applied to the $C_{18}$ column and washed with $H_2O$ several times to remove the sugars and water soluble pigments. Neochlorogenic acid, chlorogenic acid and 4-caffeoylquinic acid were eluted with 10% MeOH, and rhamosyl isoorientin was eluted with 30% MeOH, but maysin was eluted with 50% MeOH from the $C_18$ open column. Collected fractions were analyzed with HPLC by using revers-phase Ultras-phere $C_{18}$ column (4.6$\times$250mm, 5$\mu\textrm{m}$) and $H_2$O (10% MeOH containing 0.1% $H_3$${PO}_4$)/MeOH (100% MeOH containing 0.1% H$_3$PO$_4$) linear gradient from 20% to 90% MeOH for 35 minutes, a flow rate of 1 $m\ell$/min and detection at 340nm. The selected fractions were concentrated and applied to the silicic acid column. Maysin was eluted with 500$m\ell$ of 100% ethyl acetate from the silicic acid column for the first purification, and the purity of collected fractions was about 75%, but the purity from the second purification with the Cis column (1/2 $\times$ 43") was greater than 95%. FAB-MS spectral data was obtained with VG7O-VSEQ VG analytical fast atom bombardment mass (UK). $^1$H-NMR and $^{13}$ C-NMR data were obtained with Bruker DPX 400 MHz NMR spectrometers (German) in DMSO-d$_{6}$ at 400 and 100 MHz, respectively.vely.

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Effect of Human Follicular Fluid and Bovine Oviductal Tissue Extract on the Mouse Oocyte-Cumulus Complex (사람 난포액과 소의 수란관 조직추출액이 생쥐 난구세포에 미치는 영향)

  • 홍민정;김지수;심명선;김해권
    • Development and Reproduction
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    • v.6 no.2
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    • pp.97-104
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    • 2002
  • In most mammals, mature oocyte-cumulus complexer(OCCs) ovulate into the oviduct where fertilization by sperm takes place. However, the complex that fail to fertilize eventually undergoes degeneration while they reside in the oviduct. Yet there is no blown mechanism how both oocyte and cumulus cells degenerate. Using human follicular fluid (hFF), bovine oviductal tissue extract (BOX) and mouse OCC, the present study aimed to find how the oviduct influence the viability of the oocyte and cumulus cells in vitro. There was no difference of oocyte maturation rate between the control and BOX-treated groups. However, there was a significant difference in the survival of cumulus cells between two groups. Cumulus cells cultured in the presence of hFF alone underwent initially expansion and then they formed monolayer in the culture dish. Even after 72 hr, they proliferated well and showed fibroblast-like morphology. Cumulus cells cultured in the presence of both hFF and BOX also expanded after 24 hr, however, after 72 hr culture, they eventually detached and degenerated. Cumulus cells cultured in the BOX alone gave a similar drastic result. When the cumulus cells cultured in the presence of BOX were stained with DAPI, their nuclei showed partial condensation and fragmentation. After detailed analysis of these cells by TUNEL assay, many nuclei of them exhibited well stained spots indicating the signs of apoptosis. Based upon these observations, it is suggested that BOX might possess a factor that leads mouse cumulus cells to undergo apoptosis in vitro.

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Sodium Salicylate(NaSaL) Induces Apoptosis of NCI-H1299 Lung Carcinoma Cells via Activation Caspase-3 Protease (NCI-H1299 폐암 세포주에서 Caspase-3 Protease 활성을 통한 Sodium Salicylate(NaSaL)의 세포고사)

  • Shim, Hyeok;Yang, Sei-Hoon;Bak, Sang-Myeon;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.5
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    • pp.485-496
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    • 2002
  • Background : Nonsteroidal anti-inflammatory drugs (NSAIDs) are useful in the chemoprevention of colon cancers. Continuous NSAID use results in a 40% to 50% reduction in the relative risk of colorectal cancer. The precise mechanism by which NSAIDs prevent and/or cause the regression of colorectal tumors is not known. Some investigators have reported that certain NSAIDs induce apoptosis and alter the expression of the cell cycle regulatory genes in some carcinoma cells when administered at a relatively high concentration. However, the possibility of NSAIDs application as chemopreventive agents in lung cancers remains to be elucidated. To address this question, the human lung cancer cell line NCI-H1299 was used to investigate whether or not NSAIDs might induce the apoptotic death of NCI-H1299 cells. Methods : A viability test was carried out using a MTT assay. Apoptosis was measured by flow cytometric analysis and unclear staining(DAPI). The talytic activity of the caspase family was measured by the fluirigenic cleavage of biosubstrates. To define the mechanical basis of apoptosis, western blot was performed to analyze the expression of the death substrates(PARP and ICAD). Results : NaSaL significantly decreased the viability of the NCI-H1299 cells, which was revealed as apoptosis characterized by an increase in the $subG_0/G_1$ population and unclear fragmentation. The catalytic activity of caspase-3 protease began to increase after 24 Hr and reached a peak 30 Hr after treatment with 10 mM NaSaL. In contrast, caspase-6, 8, and 9 proteases did not have a significantly altered enzymatic activity. Consistent with activation of caspase-3 protease, NaSaL induced the cleavage of the protease biosubstrate. Conclusion : NaSaL induces the apoptotic death of NCI-H1299 human lung cancer cells via the activation of caspase-3 protease.

Reconsideration of Rare and Endangered Plant Species in Korea Based on the IUCN Red List Categories (IUCN 적색목록 기준에 의한 환경부 멸종위기 야생식물종에 대한 평가)

  • Chang, Chin-Sung;Lee, Heung-Soo;Park, Tae-Yoon;Kim, Hui
    • The Korean Journal of Ecology
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    • v.28 no.5
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    • pp.305-320
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    • 2005
  • Recently 64 species in Korea have been ranked as rare and endangered taxa by the Ministry of Environment using two categories, I and II. The original threat categories produced by the Ministry of Environment were developed to provide a standard for specifying animals and plants in danger of extinction and has been influential sources of information used in species conservation in Korea. However, the criteria by Ministry of Environment were applied to the whole taxa only by regional boundaries, especially in South Korea, rather than international context, and it also lacked an explicit framework that was necessary to ensure repeatability among taxa because of the absence of quantitative criteria to measure the likelihood of extinction. The World Conservation Union (IUCN) has developed quantitative criteria for assessing the conservation status of species. The threatened species categories, the 2000 IUCN Red List, proposed by SSC (Species Survival Commission) of IUCN have become widely recognized internationally. Details of threatened Korean plants, identified by applying the IUCN threat categories and definitions, were listed and analyzed. The number of species identified as threatened was only 34 out of 64 taxa (48.4%), while the rest of taxa were rejected from the original lists. Many of the species (51.6%, 33 taxa) excluded from the original list proposed by Ministry of Environment do not qualify as Critically Endangered, Endangered or Vulnerable because these taxa were widely distributed either in Japan or in China/far eastern Russia and there is no evidence of substantial decline in these countries. An evaluation of taxa in Korea has been carried out only based on subjective views and qualitative data, rather than quantitative scientific data, such as rates of decline, distribution range size, population size, and risk of extinction. Therefore, the national lists undermine the credibility of threatened species lists and invite misuse, which have been raised by other cases, qualitative estimate of risk, political influence, uneven taxonomic or geographical coverage. The increasing emphasis on international responsibilities means that global scale is becoming more significant. The current listings by Environment of Ministry of Korea should be challenged, and the government should seek to facilitate the resolution of disagreements. Especially the list should be flexible enough to handle uncertainty and also incorporates detailed, quantitative data. It is suggested that the highest priorities for the Red List should be given to endemic species in Korea first. After setting up the list of endemic species to Korea, quantitative data on population size and structure, distributional range, rated of decline, and habitat fragmentation should be collected as one of long term projects for the Red list categories. Transparency and accountability are the most important key factors. Also, species assessors are named and data sources referenced are required for the future objective evaluations on Korean plant taxa.

Effect of Natural Tenderizers or Phosphates on Quality Improvement of the Low-grade Seasoned Hanwoo Ribs (천연연화제 및 인산염의 첨가가 저급양념한우갈비의 품질개선에 미치는 효과)

  • Kim, K. J.;Min, J. S.;Lee, S. O.;Jang, A.;Jang, S. H.;Cheon, Y. H.;Lee, M.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.309-318
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    • 2003
  • In order to improve the quality of low-grade seasoned ribs, kiwi powder, pear powder and phosphates were studied. To seasoned ribs, 0.1%, 0.3% or 0.5% of kiwi powder was added and their MFI(Myofibrillar Fragmentation Index) values were 341.4, 368.3 and 405.1, respectively. As the amount of kiwi increased, MFI value increased(p<0.001). Also, when 0.5%, 1.0% or 3.0% of pear powder was added to seasoned ribs, their MFI values were increased as the addition levels of pear powder increased. As the amount of kiwi and pear powder were increased, WHC(Water Holding Capacity) decreased(p<0.001). On the other hand, as the amount of phosphates added increased, WHC increased. In drip loss, as the amount of kiwi and pear powder increased, it increased. However, drip loss was decreased as the amount of phosphates increased. For meat color, ‘L’ values of pear treatment groups were similar to those of control and ‘a’ values were higher in the treatment groups of 0.1${\sim}$0.3% kiwi powder, 0.5${\sim}$1.0% pear powder and 0.3${\sim}$0.5% phosphate than those of control. In sensory evaluation, treatment group of 0.3% kiwi powder and 0.5% phosphate showed the highest values in juiciness, tenderness and acceptability among the treatments(p<0.01).

Protective Effect of Kefir Grain Against Dextran Sodium Sulfate-Induced Colitis in Rats (Dextran Sodium Sulfate로 대장염을 유도한 흰쥐에서 캐피어 원말의 장보호 효과)

  • Ko, Young-Eun;Kim, Mi-Kyoung;Cho, Han-Young;Lee, In-Young;Ly, Sun-Yung
    • Journal of Nutrition and Health
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    • v.41 no.5
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    • pp.391-401
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    • 2008
  • Probiotics have emerged as a potential treatment modality for numerous gastrointestinal disorders, including IBD. However, few probiotics have undergone appropriate preclinical screening in vivo. Kefir is considered a probiotic, benefiting the host through its effects in the intestinal tract. Despite numerous studies examining the action of probiotics on the host organism, few have analyzed the effects on intestinal environment. We assessed the protective effect of kefir for three weeks before inducing colitis with 2% dextran sodium sulfate for five days. The DSS loads were similar in all DSS treatment group. The results of the experiment are as follows. Food intake and FER of experimental groups were not significantly different each other, but water consumption tended to be higher in all DSS treatment groups as compared with the normal control. And visual inspection of feces revealed mild diarrhea in rat given 2% DSS. The anti-inflammatory activity of kefir was determined by myeloperoxidase activity during the DSS treatment, and there was no significant difference in any group. The levels of thiobarbituric acid reactive substances (TBARS) as a colonic lipid peroxidation were significantly lower in the kefir intake groups than in rats treated with 2% DSS alone. The DNA % in tail and tail moment values as a DNA damage level of the blood lymphocytes in kefir intake groups tended to be lower than 2% DSS treatment alone, especially tail lengths were significantly diminished. According to the colonic histopathological assay, there were a severe inflammation of lamina propria and submucosa and mild edema in mucosa and sub mucosa in DSS alone treated group. We found a slight regenerative change in kefir treatment groups. In our experiments, this means that ulcerative colitis related to oxidative injury might be prevented by kefir as a probiotic. Further studies of the potential benefits of kefir as a probiotic in inflammatory condition are encouraged.

Comparison of Northgate SD-3 and Modulith SLX Lithotriptors: Treatment Results with 2,000 Renal and Ureteral Stones (Northgate SD-3와 Modulith SLX 쇄석기를 이용한 체외충격파쇄석술의 비교분석: 단일 신결석 및 요로결석 2,000례의 치료결과 분석)

  • Lee, Jun-Young;Jung, Hee-Chang;Moon, Ki-Hak;Cho, Chul-Kyu;Park, Tong-Choon
    • Journal of Yeungnam Medical Science
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    • v.16 no.1
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    • pp.85-93
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    • 1999
  • Some reports have shown a decreased effectiveness of extracorporeal shock wave lithotripsy(ESWL) with newer lithotriptors. We compared the treatment results of ESWL with a second generation Northgate SD-3 and a third generation Modulith SLX device. A total of 2,000 patients underwent ESWL treatments for single urinary calculus between September, 1988 and July, 1998. 1,241 patients were treated with Northgate SD-3 between September, 1988 and December, 1995. And 759 patients were treated with Modulith SLX between January, 1996 and July 1998. The treatment results were compared using chi-square test to determine statistical significance. The overall success rate, success rate according to the location and size, the mean number of sessions, complication rate and retreatment rate were calculated, according to lithotriptor. The overall success rate was 90.6% with Northgate SD-3 and 89.1% with Modulith SLX. With Northgate SD-3 and Modulith SLX, the success rate according to the location was 91.0%(579/636) and 88.1%(236/268) in kidney: 93.2%(517/555) and 89.9%(258/287) in upper ureter: 83.3%10/12) and 94.4%167/177) in middle and lower ureter: 47.4%(18/38) and 55. 6%(15/27) in staghorn stone, respectively. The success rate according to the size of stone with Northgate SD-3 and Modulith SLX for stones with the size under 10mm was 96.1%(612/637) and 93.1%(470/505); from 11mm to 20mm was 87.3%(421/482) and 86.4%(165/191); from 21mm to 30mm, 77.5%(62/80) and 67.5%(23/34): and for stones larger than 31mm was 69%(29/42) and 62.1%(18/29), respectively. Mean number of sessions for successful fragmentation was 1.21 and 1.69, respectively with Northgate SD-3 and Modulith SLX. Retreatment rate was 16.7% and 17.5%, respectively. The complications after treatment were severe pain(6.2% with Northgate SD-3 vs. 2.0% with Modulith SLX), steinstrasse(3.4% vs. 1.9%), fever(1.2% vs. 0.5%) and perirenal hematoma(0.2% vs. 0%) in order of frequency. There was no significant difference in the effectiveness of Northgate SD-3 and Modulith SLX. However, a statistically significant difference was observed between the two lithotriptors. We concluded that ESWL with Modulith SLX is more safe compared to Northgate SD-3.

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BCR/ABL mRNA Targeting Small Interfering RNA Effects on Proliferation and Apoptosis in Chronic Myeloid Leukemia

  • Zhu, Xi-Shan;Lin, Zi-Ying;Du, Jing;Cao, Guang-Xin;Liu, Gang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.4773-4780
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    • 2014
  • Background: To investigate the effects of small interference RNA (siRNA) targeting BCR/ABL mRNA on proliferation and apoptosis in the K562 human chronic myeloid leukemia (CML) cell line and to provide a theoretical rationale and experimental evidence for its potential clinical application for anti-CML treatment. Materials and Methods: The gene sequence for BCR/ABL mRNA was found from the GeneBank. The target gene site on the BCR/ABL mRNA were selected according to Max-Planck-Institute (MPI) and rational siRNA design rules, the secondary structure of the candidate targeted mRNA was predicted, the relevant thermodynamic parameters were analyzed, and the targeted gene sequences were compared with BLAST to eliminate any sequences with significant homology. Inhibition of proliferation was evaluated by MTT assay and colony-formation inhibiting test. Apoptosis was determined by flow cytometry (FCM) and the morphology of apoptotic cells was identified by Giemsa-Wright staining. Western blotting was used to analyze the expression of BCR/ABL fusion protein in K562 cells after siRNA treatment. Results: The mRNA local secondary structure calculated by RNA structure software, and the optimal design of specific siRNA were contributed by bioinformatics rules. Five sequences of BCR/ABL siRNAs were designed and synthesized in vitro. Three sequences, siRNA1384, siRNA1276 and siRNA1786, which showed the most effective inhibition of K562 cell growth, were identified among the five candidate siRNAs, with a cell proliferative inhibitory rate nearly 50% after exposure to 12.5nmol/L~50nmol/L siRNA1384 for 24,48 and 72 hours. The 50% inhibitory concentrations ($IC_{50}$) of siRNA1384, siRNA1276 and siRNA1786 for 24hours were 46.6 nmol/L, 59.3 nmol/L and 62.6 nmol/L, respectively, and 65.668 nmol/L, 76.6 nmol/L, 74.4 nmol/L for 72 hours. The colony-formation inhibiting test also indicated that, compared with control, cell growth of siRNA treated group was inhibited. FCM results showed that the rate of cell apoptosis increased 24 hours after transfecting siRNA. The results of annexinV/PI staining indicated that the rate of apoptosis imcreased (1.53%, 15.3%, 64.5%, 57.5% and 21.5%) following treamtne with siRNAs (siRNA34, siRNA372, siRNA1384, siRNA1276 and siRNA1786). Morphological analysis showed td typical morphologic changes of apoptosis such as shrunken, fragmentation nucleus as well as "apoptotic bodies" after K562 cell exposure to siRNA. Western blot analysis showed that BCR/ABL protein was reduced sharply after a single dose of 50nmol/L siRNA transfection. Conclusions: Proliferation of K562 cells was remarkbly inhibited by siRNAs (siRNA1384, siRNA1276 and siRNA1786) in a concentration-dependent manner in vitro, with effective induction of apoptosis at a concentration of 50 nmol/L. One anti-leukemia mechanism in K562 cells appeared that BCR/ABL targeted protein was highly down-regulated. The siRNAs (siRNA1384, siRNA1276 and siRNA1786) may prove valuable in the treatment of CML.

Vegetation Structure Characteristics and Management Plan of Mulgeun Fish Shelter Forest in the Southern Coast (남해안 물건리 방조어부림의 식생구조 특성 및 관리방안)

  • Lee, Soo-Dong;Kim, Mi-Jeong;Kang, Hyun-Kyung
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.34 no.1
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    • pp.118-128
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    • 2016
  • The purpose of this study is to present efficient methods of preserving and managing the fish shelter forest in Mulgun-ri on the southern coast of Korea on the basis of its humanistic, sociological and ecological characteristics. The study object is Korean natural monument No. 150, which is presumed to have been forested by descendants of Jeonju Lee Family who settled there, and village rituals are held every October to pray for the peace of the village. The forest is managed by Namhae-gun as a historical and cultural resource as well as its disaster-preventing, economic, and environmental and ecological functions. The linear form of the area is $23,962.6m^2$ and farmland(48.5%) and urbanization area(38.2%) are extensively located in its periphery area. Actual vegetation was sub-classified into three types of land according to use pressure and whether or not damage was done: land where its stratification was formed; land where it was restored, and the land where it was damaged. Plant communities were sub-classified into Aphananthe aspera community(I) and Zelkova serrata community(II) which had a low use pressure; Z. serrata-Chionanthus retusa-A. aspera community(III) and A. aspera-Z. serrata community(IV) which had a high use pressure; and Celtis sinensis-A. aspera community(V) whose underlayer was damaged by use. Fragmentation of the forest is under way and its inside vegetation growth is hampered due to the installation of traffic and resting facilities such as the through roads costal roads, wooden-deck walkways, parking lots, washstands, etc. As a restoration management plan for this, the following were required: an establishment of preferred restoration area; a selection of restoration vegetation species; and an appropriate restoration method. The damaged area($7,868.2m^2$) will have to be set up as the preferred restoration area; seedlings of restored vegetation species should be raised with dominant species within the forest(i.e., Z. serrata, A. aspera, C. sinensis, and C. retusa) as their 'mother trees' for the benefit of for the next-generation forest; and sub-tree and shrub layer should be complementarily planted with 5 and 115 trees(unit $100m^2$) respectively to facilitate the formation of a multi-layered vegetation structure. In addition, resting facilities scattered inside the forest should be demolished; and indiscriminate use of them should be controlled; management and monitoring should be carried out so that the area can be preserved and restored as a deciduous broad-leaved forest.