• Title/Summary/Keyword: membrane chemistry

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A Novel Chenodeoxycholic Derivative HS-1200 Induces Apoptosis in Human HT-29 Colon Cancer Cells (인체 대장암 세포주(HT-29)에서 담즙산 합성유도체(HS-1200)의 세포 사망 기전)

  • Oh Sin Geun;Yang Kwang Mo;Hur Won Joo;Yoo Young Hyun;Suh Hong Suk;Lee Hyung Sik
    • Radiation Oncology Journal
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    • v.20 no.4
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    • pp.367-374
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    • 2002
  • Purpose : To investigate the growth inhibitory effects, and the underlying mechanism of human colon cancer cell (HT-29) death, induced by a new synthetic bile acid derivative (HS-1200). Materials and Methods : Human colon cancer cells (HT-29), in exponential growth phase, were treated with various concentrations of a new synthetic bile acid derivative (HS-1200). The growth inhibitory effects on HT-29 cells were examined using a frypan blue exclusion assay. The extent of apoptosis was determined using agarose gel electrophoresis, TUNEL assays and Hoechst staining. The apoptotic cell death was also confirmed by Western blotting of PARP, caspase-3 and DNA fragmentation factor (DFF) analysis. To investigate the involvement of mitochondria, we employed immunofluorescent staining of cytochrome c and mitochondrial membrane potential analyses. Results : The dose required for the half maximal inhibition $(IC_{50})$ of the HT-29 cell growth was $100\~150\;{\mu}M$ of HS-1200. Several changes, associated with the apoptosis of the HT-29 cells, were reveal by the agarose gel eletrophoresis, TUNEL assays and Hoechst staining, following their treatment with $100\;{\mu}M$ of HS-1200. HS-1200 treatment also induced caspase-3, PARP and DFF degradations, and the western blotting showed the processed caspase-3 p20, PARP p85 and DFF p30 and p11 cleaved products. Mitochondrial events were also demonstrated. The cytochrome c staining indicated that cytochrome c had been released from the mitochondria in the HS-1200 treated cells. The mitochondrial membrane potential $(\Delta\Psi_m)$ was also prominently decreased in the HS-1200 treated cells. Conclusion : These findings suggest that the HS-1200 - induced apoptosis of human colon cancer cells (HT-29) is mediated via caspase and mitochondrial pathways.

Angiotensin-Converting Enzyme Inhibitory Activity of Enzymatic Hydrolysates of Crassostrea gigas (Oyster) (굴 효소 가수분해물의 angiotensin converting enzyme 저해작용)

  • Do, Hyung-Joo;Park, Hye-Jin;Kim, Ok-Ju;Kim, Andre;Choi, Yeung-Joon;Choung, Se-Young;Ha, Jong-Myung
    • Journal of Life Science
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    • v.22 no.2
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    • pp.220-225
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    • 2012
  • The peptides of enzymatic hydrolysates from oyster were determined by inhibitory activity against angiotensin-converting enzyme. The ACE inhibitory activity of enzymatic oyster hydrolysates increases with hydrolysis time. Among enzymatic oyster hydrolysates, oyster hydrolysates incubated with Protamex showed the best ACE inhibitory activity after 10 h. Hydrolysates were filtered through a HiSep ultrafiltration membrane (M.W. cut-off 30 kDa, 10 kDa) to obtain the peptide fractions with ACE inhibition activity. These fractions were applied to an HPLC column (watchers 120 ODS-AP $250{\times}4.6$ ($5{\mu}m$)). Six active fractions were collected and the range of ACE inhibition was from 29.56 to 85.85%. Peptide was purified from fraction B, showing the highest ACE inhibitory activity, and its sequence was Leu-Gln-Pro. These results suggest that PEH may be beneficial for developing antihypertensive food and drug.

Physiological Roles of Phospholipase Cγ and Its Mutations in Human Disease (Phospholipase Cγ의 생리적 기능과 질병과 연관된 돌연변이)

  • Jang, Hyun-Jun;Choi, Jang Hyun;Chang, Jong-Soo
    • Journal of Life Science
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    • v.30 no.9
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    • pp.826-833
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    • 2020
  • Phospholipase C gamma (PLCγ) has critical roles in receptor tyrosine kinase- and non-receptor tyrosine kinase-mediated cellular signaling relating to the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] to produce inositol 1,4,5 trisphosphate (IP3) and diacylglycerol (DAG), which promote protein kinase C (PKC) and Ca2+ signaling to their downstream cellular targets. PLCγ has two isozymes called PLCγ1 and PLCγ2, which control cell growth and differentiation. In addition to catalytically active X- and Y-domains, both isotypes contain two Src homology 2 (SH2) domains and an SH3 domain for protein-protein interaction when the cells are activated by ligand stimulation. PLCγ also contains two pleckstrin homology (PH) domains for membrane-associated phosphoinositide binding and protein-protein interactions. While PLCγ1 is widely expressed and appears to regulate intracellular signaling in many tissues, PLCγ2 expression is restricted to cells of hematopoietic systems and seems to play a role in the regulation of immune response. A distinct mechanism for PLCγ activation is linked to an increase in phosphorylation of specific tyrosine residue, Y783. Recent studies have demonstrated that PLCγ mutations are closely related to cancer, immune disease, and brain disorders. Our review focused on the physiological roles of PLCγ by means of its structure and enzyme activity and the pathological functions of PLCγ via mutational analysis obtained from various human diseases and PLCγ knockout mice.

Physicochemical Heterogeneity of the Supply Water Quality depending on Seasonal Changes in the Taejon Area, Korea (대전지역 상수도 물의 계절변화에 따른 물리화학적 불균질)

  • 이현구;이찬희;서호택
    • Economic and Environmental Geology
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    • v.33 no.6
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    • pp.505-517
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    • 2000
  • This study was undertaken to provide a drinking water quality on the basis of physicochemical properties. In this study, the 25 samples of supply waters of the Taejon area were sampled twice (February and August in 1999). Hydrochemistry of the supply water belongs to the $Ca^{2+}$-${HCO_3}^{-1}$ type, whereas the supply water was characterized by the relatively significant enrichment of ${Ca}^{2+}$, ${Na}^{2+}$, ${K}^{2+}$, ${Cl}^{2+}$ ions and heavy metals compared to the original water from the Daecheong lake. Generally, the supply water has a mean values for $10.7^{\circ}C$ of temperature, 6.86 of pH, -12 mV of Eh, 88 ${\mu}S$/cm of EC and 70.379 mg/l of TDS in February, whereas the waters of the same sites in August are a slightly high temperature ($26.1^{\circ}C$), TDS (78.069 mg/l) and extremely high EC (442 ${\mu}S$/cm) value. These values are similar with physicochemical properties of the original lake water depending on the seasonal differences. Results of speciation calculation indicate that potentially toxic ions might exist mainly in the forms of free metal (${Cu}^{2+}$ or ${Zn}^{2+}$) and a small amount of ${CO_3}^{2-}$and ${OH}^{-}$in the supply water. The water seemed to be in equilibrium with kaolinite field of the normal stability diagrams for the natural water. Based on enrichment parameter of the supply water normalized by original lake water composition, the average value of those parameter can be calculated with nearly 1.00, but the those values for Cu+Zn possible source of decrepit pipe lines are 126.75 in February and 115.63 in August samples. The parameter values varied with sampling sites, however, do not exceed by chemistry of drinking water standard. Solid compounds remained on the membrane filter papers after filtration are adhered to pale yellow or yellowish brown colored dissolved solids and precipitates, which are coated by 0.02 to 0.35 mm thick per 500 ml with colloidal particles of about 1 to 2${\mu}m$ size. The particles are mainly Fe-Cu-Zn compounds and partly detected to Mn and Pb.

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Flavonoids from Orostachys Japonicus A. Berger Induces Caspase-dependent Apoptosis at Least Partly through Activation of p38 MAPK Pathway in U937 Human Leukemic Cells

  • Lee, Won Sup;Yun, Jeong Won;Nagappan, Arulkumar;Jung, Ji Hyun;Yi, Sang Mi;Kim, Dong Hoon;Kim, Hye Jung;Kim, GonSup;Ryu, Chung Ho;Shin, Sung Chul;Hong, Soon Chan;Choi, Yung Hyun;Jung, Jin-Myung
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.465-469
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    • 2015
  • Background: Orostachys japonicus A. Berger (A. Berger) is commonly used as a folk remedy for cancer therapy. However, the mechanisms of its anti-cancer activity are poorly investigated in human cancer cells. In this study, we investigated whether flavonoids extracted from Orostachys japonicus A. Berger (FEOJ) might have anticancer effects in human leukemia cells, focusing on cell death mechanisms. Materials and Methods: U937 human leukemic cancer cells were used. Results: FEOJ induced apoptosis in a dose-dependent manner in human U937 cancer cells. Flow cytometry revealed significant accumulation of cells with sub-G1 DNA content at the concentrations of $200{\mu}g/mL$ and $400{\mu}g/mL$. FEOJ-induced apoptosis was caspase-dependent through loss of mitochondrial membrane potential (MMP, ${\Delta}{\Psi}m$) in human U937 cancer cells, which might be associated with suppression of Bcl-2 and XIAP proteins. FEOJ induced the p38 MAPK signaling pathway, playing at least in part an important role in FEOJ-induced apoptosis. Conclusions: This study suggested that FEOJ may induce caspase-dependent apoptosis in human leukemic cells by regulating MMP (${\Delta}{\Psi}m$) through suppressing Bcl-2 and X-IAP. In addition, the results indicated that upstream p38 MAPK signaling regulates the apoptotic effect of FEOJ. This study provides evidence that FEOJ might have anti-cancer potential for human leukemic cells.

Anti-metastatic Effects on B16F10 Melanoma Cells of Extracts and Two Prenylated Xanthones Isolated from Maclura amboinensis Bl. Roots

  • Siripong, Pongpun;Rassamee, Kitiya;Piyaviriyakul, Suratsawadee;Yahuafai, Jantana;Kanokmedhakul, Kwanjai
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3519-3528
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    • 2012
  • Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.

Solvent Tolerant Bacteria and Their Potential Use (유기용매 내성 세균과 이용가능성)

  • Joo, Woo Hong
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1458-1469
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    • 2015
  • Many organic solvent-tolerant bacteria have been isolated from all environments such as soil, waste-water, even deep sea after first isolation report of organic solvent-tolerant bacterium. Most organic solvent- tolerant isolates have been determined to be Gram-negative bacteria, because Gram-negative bacteria have inherent tolerance property toward hostile organic solvents more than Gram-positive bacteria. The mechanisms of organic solvent tolerance have been elucidated extensively using mainly organic solvent-tolerant Gram-negative bacteria. The solvent-tolerance mechanisms in Gram-positive bacteria can be found in comparatively recent research. Organic solvents exhibited different toxicity depending on the solvent, and the tolerance levels of organic solvent-tolerant bacteria toward organic solvents were also highly changeable among species and strains. Therefore, organic solvent-tolerant bacteria could coped with solvent toxicity and adapted to solvent stress through the multifactorial and multigenic adaptative strategies. They could be survived even in the hyper concentrations of organic solvents by mechanisms which include: changes in cell morphology and cell behaviour, cell surface modifications, cell membrane adaptations, solvent excretion pumps, chaperones and anti-oxidative response. The aim of this work is to review the representative solvent tolerant bacteria and the adaptative and tolerance strategies toward organic solvents in organic solvent-tolerant bacteria, and their potential industrial and environmental impact.

Heterologous Expression of Fission Yeast Heavy Metal Transporter, SpHMT-1, Confer Tolerance to Cadmium in Cytosolic Phytochelatin-Deficient Saccharomyces cerevisiae (분열효모 SpHMT1을 세포질 파이토킬레이트를 생성하지 않는 효모에서 발현으로 인한 카드뮴에 대한 저항성 증가)

  • Lee, Sang-Man
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1685-1689
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    • 2009
  • Phytochelatins (PCs) are small polypeptides synthesized by PC synthase (PCS). They are present in various living organisms including plants, fission yeast, and some animals. The presumed function of PCs is the sequestration of cytosolic toxic heavy metals like cadmium (Cd) into the vacuoles via vacuolar membrane localized heavy metal tolerance factor 1 (HMT-1). HMT-1 was first identified in fission yeast (SpHMT-1), and later in Caenorhabdtis (CeHMT-1). Recently, its homolog has also been found in PC-deficient Drosophila (DmHMT-1), and this homolog has been shown to be involved in Cd detoxification, as confirmed by the heterologous expression of DmHMT-1 in fission yeast. Therefore, the dependence of HMT-1 on PC in Cd detoxification should be re-evaluated. I heterologously expressed SpHMT-1 in cytosolic PC-deficient yeast, Saccharomycea cerevisiae, to understand the dependence of HMT-1 on PC. Yeast cells expressing SpHMT-1 showed increased tolerance to Cd compared with control cells. This result indicates that SpHMT-1 is not strictly correlated with PC production on its function. Moreover, yeast cells expressing SpHMT-1 showed increased tolerance to exogenously applied glutathione (GSH) compared with control cells, and the tolerance to Cd was further increased by exogenously applied GSH, while tolerance in control cells was not. These results indicate that the function of SpHMT-1 in Cd detoxification does not depend on PCs only, and suggest that SpHMT-1 may sequester cytosolic GSH-Cd complexes into the vacuole.

Study on the Tensile Strength of Oxidative Permanent Dyed Hair (산화형 영구 염모제로 처리한 모발의 인장강도 특성에 관한 연구)

  • Lee, Gui-Young;Chang, Byung-Soo
    • Applied Microscopy
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    • v.38 no.4
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    • pp.339-345
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    • 2008
  • In this study, we compared the stress and tensile strength of the hair treated with oxidative permanent dye with those of virgin hair. We investigated the fine structure of the hair section after the tensile test using scanning electron microscopy. The tensile strength of the virgin hair was measured as $14.66\;g/cm^2$, tensile energy was $108\;erg/cm^2$, and the maximum stress was 146.64g. Those of the dyed hair were $13.69\;g/cm^2$, $89.62\;erg/cm^2$ and 136.90 g, respectively. The differences in the tensile strength, the tensile energy and the maximum stress were $-0.97\;g/cm^2$, $-18.38\;erg/cm^2$, -9.74 g, respectively, which showed that the dyed hair had less elasticity and strength than the virgin hair. In the scanning electron microscopy investigation of the damaged hair after the tensile test, lift-off of the cuticle outer layer were shown in both virgin hair and dyed hair, which was more severe in the dyed hair than the virgin hair. Adjacent cuticular cells of the cuticle layer were separated by the destruction of intercellular membrane complex. The macrofibrils were exposed and separated from the cortex torn by tensile strength.

Manufacture and Evaluation of Small Size PEMFC Stack Using Carbon Composite Bipolar Plate (탄소복합소재 분리판을 이용한 소형 고분자전해질 연료전지 스택 제작 및 성능분석)

  • Han, C.;Choi, M.;Lee, J.J.;Lee, J.Y.;Kim, I.T.;An, J.C.;Shim, J.;Lee, H.K.
    • Journal of the Korean Electrochemical Society
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    • v.13 no.2
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    • pp.81-88
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    • 2010
  • Small size polymer electrolyte membrane fuel cell (PEMFC) stacks were prepared using carbon composite and graphite bipolar plates and their performances were evaluated on reactant gas and operating time. In comparison to single cell and stack, it was identified that home-made bipolar plate was well-designed to maximize stack performance as high as that of single cell. During long-term operation, the performances of stacks using two different kinds of bipolar plates were compared. The decrease of performance in both stacks was accelerated with increasing load current. It was observed from stack test that the stack performance using carbon composite bipolar plate was very similar to that using graphite bipolar plate.