• Title/Summary/Keyword: Bio-analysis

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Induction of Apoptosis by Pectenotoxin-2 Isolated from Marine Sponges in U937 Human Leukemic Cells (인체 혈구암세포 U937에서 해양해면동물에서 추출된 Pectenotoxin-2에 의한 Apoptosis의 유발에 관한 연구)

  • Shin, Dong Yeok;Kang, Ho Sung;Bae, Song-Ja;Jung, Jee H.;Choi, Yung Hyun
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.63-70
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    • 2006
  • Natural product compounds are the source of numerous therapeutic agents. The marine environment produces natural products from a variety of structural classes exhibiting activity against numerous disease targets including anticancer agents. Among these, pectenotoxin-2 (PTX-2), which was first identified as a cytotoxic entity in marine sponges, which depolymerizes actin filaments, was found to be highly effective and more potent to activate an intrinsic pathway of apoptosis in p53-deficient tumor cells compared to those with functional p53 both in vitro and in vivo. However, the anti-proliferative mechanism of the compound at non-cytotoxic concentrations has not yet been explored. In the current study, we sought to investigate anti-proliferation and apoptosis of PTX-2 against U937 human leukemic cells and its underlying molecular mechanism. Exposure of U937 cells to PTX-2 resulted in growth inhibition and induction of apoptosis in dose- and time-dependent manner as measured by MTT assay, fluorescent microscopy and flow cytometric analysis. The anti-proliferative effect of PTX-2 was associated with a marked increase in the expression of cyclin-dependent kinase p21 (WAF1/CIP1) mRNA which was tumor suppressor p53-independent. The increase in apoptosis was connected with a time-dependent down-regulation of anti-apoptotic Bcl-XL and inhibitor of apoptosis proteins (IAPs) family such as XIAP and cIAP-2. Though additional studies are needed, these findings suggested that PTX-2-induced inhibition of U937 cells was associated with the induction of apoptotic cell death and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of PTX-2.

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Glucocorticoid Regulation of Gene Expression in Hippocampal CA3 and Dentate Gyrus (글루코코티코이드 호르몬에 의한 뇌해마의 CA와 Dentate Gyrus 부분의 유전자 발현 변화)

  • Kim, Dong-Sub;Ahn, Soon-Cheol;Kim, Young-Jin;Park, Byoung-Keun;Ahn, Yong-Tae;Kim, Ji-Youn;Kyoji, Morita;Her, Song
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.305-311
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    • 2007
  • Glucocorticoids (GCs) alter metabolism, synaptogenesis, apoptosis, neurogenesis, and dendritic morphology in the hippocampus. To better understand how glucocorticoids regulate these aspects of hippocampal biology, we studied gene expression patterns in the CA3 (Hippocampal pyramidal cell field CA3) and dentate gyrus (DG). Litter-matched Lewis inbred rats treated for 20 days with either 9.5 mg per day sustained-release corticosterone or placebo pellets were compared with high-density oligonucleotide microarray analysis (Rat Neurobiology U34 Arrays, Affymetrix). In placebo-treated rats, 32 genes were expressed at greater levels in CA3 than DG, whereas 3 genes were expressed at great levels in DC than CA3. Regional differences were also apparent in corticosterone-induced changes in the hippocampal transcriptome. Six genes in CA3 and 41 genes in DC were differentially regulated by corticosterone. As per the glucocorticoid effects on gene transcription in the brain, forty three of these genes were upregulated, and 4 genes were downregulated. Genes differentially expressed in hippocampus included those for 13 neurotransmitter proteins, 5 ion channel related proteins, 4 transcription factors, 3 neurotrophic factors, 1 cytokine, 1 apoptosis related protein, and 5 genes involved in synaptogenesis. Interestingly, GCs can have suppressive effects on brain BDNF mRNA transcription, one of the neurotrophic factors. These results indicate the diversity of targets affected by chronic exposure to corticosterone and highlight important regional differences in hippocampal neurobiology.

Identification and Characterization of Alternative Promoters of the Rice MAP Kinase Gene OsBWMK1

  • Koo, Sung Cheol;Choi, Man Soo;Chun, Hyun Jin;Park, Hyeong Cheol;Kang, Chang Ho;Shim, Sang In;Chung, Jong Il;Cheong, Yong Hwa;Lee, Sang Yeol;Yun, Dae-Jin;Chung, Woo Sik;Cho, Moo Je;Kim, Min Chul
    • Molecules and Cells
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    • v.27 no.4
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    • pp.467-473
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    • 2009
  • Our previous study suggested that OsBWMK1, a gene which encodes a member of the rice MAP kinase family, generates transcript variants which show distinct expression patterns in response to environmental stresses. The transcript variants are generated by alternative splicing and by use of alternative promoters. To test whether the two alternative promoters, pOsBWMK1L (promoter for the OsBWMK1L splice variant) and pOsBWMK1S (promoter for the OsBWMK1S splice variant), are biologically functional, we analyzed transgenic plants expressing GUS fusion constructs for each promoter. Both pOsBWMK1L and pOsBWMK1S are biologically active, although the activity of pOsBWMK1S is lower than that of pOsBWMK1L. Histochemical analysis revealed that pOsBWMK1L is constitutively active in most tissues at various developmental stages in rice and Arabidopsis, whereas pOsBWMK1S activity is spatially and temporally restricted. Furthermore, the expression of pOsBWMK1S::GUS was upregulated in response to hydrogen peroxide, a plant defense signaling molecule, in both plant species. These results suggest that the differential expression of OsBWMK1 splice variants is the result of alternative promoter usage and, moreover, that the mechanisms controlling OsBWMK1 gene expression are conserved in both monocot and dicot plants.

Purification and characterization of the chitinase from Bacillus subtilis JK-56 (Bacillus subtilis JK-56이 생산하는 chitinase isozyme의 정제와 특성 규명)

  • 전홍기;김낙원;정영기
    • Journal of Life Science
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    • v.12 no.1
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    • pp.77-86
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    • 2002
  • Chitin, a $\beta$-1,4 polymer of N-acetyl-D-glucosamine, is one of the most abundant organic compounds in nature. Chitinase (EC 3.2.1.14) is an enzyme that degrades chitin to chito-oligosaccharides, diacetyl rhitobiose and N-acetyl-D-glucosamine. An extracellular chitinase-producing bacterial strain was isolated from soil and named to as Bacillus subtilis JK-56. Optimum culture condition of B. subtilis JK-56 for the production of chitinase was 1% chitin, 0.5% polypepton, 0.1% KCl, 0.05% MnS $O_4$.4$H_2O$, 37$^{\circ}C$, initial pH 7.0 and 40 hour culture time. When B. subtilis JK-56 was grown in the optimum medium, one major active band and two minor active bands were detected by native-PAGE and active staining of the gel. Among them, the major band was purified from the culture supernatant by 70% ammonium sulfate precipitation and native-PAGE with BIO-RAD Model 491 Prep-Cell and named as Chi-56A. Its molecular weight was estimated to be 53kDa monomer and the isoelectric point (pI) was pH 4.3. The pH and temperature for the optimum activity of Chi-56A were pH 6.0 and $65^{\circ}C$, respectively. Chi-56A was stable up to $65^{\circ}C$ and in alkaline region. Its $K_{m}$ value for colloidal chitin was 17.33g/L. HPLC analysis of the reaction products confirmed that Chi-56A was an exo type chitinase.e.

Development of Disposable Immunosensors for Rapid Determination of Sildenafil and Vardenafil in Functional Foods

  • Vijayaraj, Kathiresan;Lee, Jun Hyuck;Kim, Hyung Sik;Chang, Seung-Cheol
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.83-88
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    • 2017
  • We introduced disposable amperometric immunosensors for the detection of Sildenafil and Vardenafil (SDF/VDF) based on screen printed carbon electrodes. The developed immunosensors were used as a non-competitive sandwich-type enzyme immunoassay with a horseradish peroxidase label. The sensors were constructed on screen printed carbon electrodes by the simple electrochemical deposition of a reduced graphene oxide and chitosan (ErGO-CS) composite. To evaluate the sensing chemistry and optimize the sensor characteristics, a series of electrochemical experiments were carried out including electrochemical impedance spectroscopy, cyclic voltammetry and amperometry. The sensors showed a linear response to SDF/VDF concentrations in a range from 100 pg/mL to 300 ng/mL. The lower detection limit was calculated to be 55 pg/mL, the sensitivity was calculated to be $1.02{\mu}Ang/mL/cm^2$, and the sensor performance exhibited good reproducibility with a relative standard deviation (RSD) of 7.1%. The proposed sensing chemistry strategy and the sensor format can be used as a simple, cost-effective, and feasible method for the in-field analysis of SDF/VDF in functional or health supplement food samples.

A study on the Characteristic of Mask Sheets (마스크 팩 시트의 특성 연구)

  • Jang, Hye-In
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.4
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    • pp.787-798
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    • 2017
  • This is a study on characteristics according to the material of sheet-type mask packs being sold on the market. The absorption capacities of water soluble components such as purified water, 1.3-propanediol, 1.3-butylene glycol, glycerine, and hyaluronic acid are compared with that of various oils including cyclomethicone, dimethicone, phytosqualane, caprylic capryl triglyceride, grape seed oil, and macadamia nut oil. As a result, all of the water soluble components except purified water showed higher moisture absorption capacity as the viscosity increased. And in case of oil, all oil showed higher oil absorption capacity according to the viscosity. During this test, the mask sheets with the type of acetic acid fermented bio-cellulose showed 500~1,000 times or more absorption capacity on water soluble wetting agent or all oils, which is due to the fine mesh structure seen in the 5,000x enlarged photograph at surface structure. This mesh structure was well recognized on the cross section and these structural features enhance the absorption capacity of water and oil. It is also believed that largely contained water-soluble components and oils facilitate the discharge over time. In addition, since each mask sheet shows their characteristics according to their material, it is intended to be a basic research for manufacturing mask packs good for skin.

Population genetic structure of Sedum polytrichoides (Crassulaceae): Insights into barriers to gene flow (바위채송화(돌나물과)집단의 유전적 구조: 유전자 이동과 물리적 장벽에 관한 통찰)

  • Chung, Mi Yoon;Lopez-Pujol, Jordi;Chung, Myong Gi
    • Korean Journal of Plant Taxonomy
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    • v.46 no.4
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    • pp.361-370
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    • 2016
  • An area comprising Juwangsan National Park and its adjacent mountains (southeastern Korean Peninsula) is a good model system for testing the effects of physical barriers to gene flows in plant populations. We predicted that plant species consisting of isolated populations are genetically more differentiated than those that are rather continuously distributed. Most populations of Sedum polytrichoides occur in four isolated valleys, and we assessed the genetic variability and structures using twelve allozyme loci in ten populations. We also compared the present results to earlier findings pertaining to the two co-occurring herbs Hylotelephium ussuriense (${\equiv}$ Sedum ussuriense) (growing only in the four isolated valleys) and S. kamtschaticum (rather continuously distributed). We found moderate levels of within-population genetic variation in S. polytrichoides ($H_{e}=0.112$). Estimates of among-population divergence in S. polytrichoides were also moderate ($F_{ST}=0.250$) and, as expected, very similar to that of H. ussuriense (0.261) but considerably higher than the variation in S. kamtschaticum (0.165). An analysis of molecular variance (AMOVA) revealed that S. polytrichoides and H. ussuriense had higher percentages of among-valley variation (19% each) than S. kamtschaticum (4%). Most of this variation, as also indicated by the STRUCTURE program, was due to differences in genetic profiles between the two central valleys. We concluded that the genetic differences observed between species (S. kamtschaticum vs. S. polytrichoides and H. ussuriense) are mainly due to differences in their distribution within the study area.

Isolation of Stenotrophomonas rhizopilae Strain GFC09 with Ginsenoside Converting Activity and Anti-wrinkle Effects of Converted Ginsenosides (사포닌 전환 활성 Stenotrophomonas rhizopilae Strain GFC09 균주의 분리 동정 및 전환 사포닌의 주름 개선 효과)

  • Min, Jin Woo;Kim, Hye-Jin;Joo, Kwang-Sik;Kang, Hee-Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.375-382
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    • 2015
  • Ginsenosides (ginseng saponin) as the one of important pharmaceutical compounds of ginseng and is responsible for the pharmacological and biological activities. These ginsenoside produces diverse small molecules ginsenoside which have more pharmacological activities including anti-wrinkle, anti-cancer and anti-oxidant effects. In the present study, we isolated bacteria using esculin agar, to produce ${\beta}$-glucosidase, and we focused on the bio-transformation of ginsenoside. Phylogenetic tree analysis was performed by comparing the 16S rRNA sequences; we identified the strain as Stenotrophomonas rhizopilae strain GFC09. In order to determine the optimal conditions for enzyme activity, the crude enzyme was incubated with 1 mM ginsenoside $Rb_1$. Bioconversion of ginsenoside $Rb_1$ were analyzed using TLC and HPLC. The crude enzyme hydrolyzed the ginsenoside $Rb_1$ along the following pathway: LB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$ into compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. The structure of the hydrolyzed metabolites were identified by NMR. The activity screening tests showed that the conversion product induced the production of type I procollagen in a dose-dependent manner. These results suggested that hydrolyzed ginseng product containing the ginsenoside $F_2$ and compound K could be useful as an active ingredient for wrinkle-care cosmetics.

Protective Effects of Helianthus annuus Seed Extract against Chemical-Induced Neuronal Cell Death (해바라기씨 추출물의 뇌세포에 대한 사멸 보호 효과)

  • Park, Ja-Young;Woo, Sang-Uk;Heo, Jin-Chul;Lee, Sang-Han
    • Food Science and Preservation
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    • v.14 no.2
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    • pp.213-219
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    • 2007
  • To develop an anti-dementia agent with potential therapeutic value in the protection of neuronal cells, we selected a water extract of Helianthus annuus seed for analysis. We measured acetylcholinesterase inhibitory activity in the extract, and analyzed the protective effect of the extract on neuronal cell death induced by hydrogen peroxide, or amyloid ${\beta}-peptide$, of SH-SY5Y neuroblastoma cells. The result showed that the extinct exerted protective effects of 83%, 72% and 53% respectively, on cell death induced by 100M, 200M, and 500M hydrogen peroxide. Also, when 50M of amyloid ${\beta}-peptide$ was added to the cells, the extract showed a protective effect (up to 80%) on cell death. Overall, the results showed that the H. annuus extract inhibited acetylcholinesterase activity in a dose-dependent manner, and the extract also strongly protected against cell death induced by hydrogen peroxide or amyloid ${\beta}-peptide$.

Antimicrobial activity and protective effect of Geranium thunbergii against oxidative DNA damage via antioxidant effect (현초의 항산화 활성에 의한 산화적 DNA 손상 보호효과 및 항균활성)

  • Kwon, Tae-Hyung;Lee, Su-Jin;Park, Jae-Ho;Kim, Taewan;Park, Jung-Ja;Park, Nyun-Ho
    • Food Science and Preservation
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    • v.24 no.3
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    • pp.325-333
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    • 2017
  • This study aimed to investigate the various biological activities of Geranium thunbergii such as antimicrobial activity and protective effect against oxidative damage. To evaluate its antioxidant and antimicrobial activities, we first performed methanol extraction; this methanol extract was further partitioned using various solvents. And then, its antioxidant activity was measured using various assays including total phenolic content and protection against oxidative DNA damage, and antimicrobial activities were examined using minimum inhibiting concentration (MIC) test, and paper disc method. In addition, high-performance liquid chromatography was performed to analyze the major chemical components of ethyl acetate fraction. The G. thunbergii fraction with ethyl acetate exhibited higher antioxidant and antimicrobial activities than the other fractions. The results showed that G. thunbergii ethyl acetate fraction at $50{\mu}g/mL$ had strong DPPH and ABTS radical scavenging activities of 80.88% and 80.12%, respectively. In addition, the ethyl acetate fraction protected DNA from the oxidative damage induced by ferrous ion and hydroxyl radicals and showed high antimicrobial activity with diameter of inhibition zones ranging from 13.33 to 15.67 mm. High-performance liquid chromatography analysis revealed the major phenolic compounds of G. thunbergii to be ellagic acid and gallic acid. These results suggest that G. thunbergii might protect DNA against oxidative stress induced by reactive oxygen species and can be utilized as a natural source of antioxidant and antimicrobial agent in the food industry.