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Protein Profiles Associated with Anoikis Resistance of Metastatic MDA-MB-231 Breast Cancer Cells

  • Akekawatchai, Chareeporn;Roytrakul, Sittiruk;Kittisenachai, Suthathip;Isarankura-Na-Ayudhya, Patcharee;Jitrapakdee, Sarawut
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.581-590
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    • 2016
  • Resistance to anoikis, a cell-detachment induced apoptosis, is one of the malignant phenotypes which support tumor metastasis. Molecular mechanisms underlying the establishment of this phenotype require further investigation. This study aimed at exploring protein expression profiles associated with anoikis resistance of a metastatic breast cancer cell. Cell survival of suspension cultures of non-metastatic MCF-7 and metastatic MDA-MB-231 cells were compared with their adherent cultures. Trypan blue exclusion assays demonstrated a significantly higher percentage of viable cells in MDA-MB-231 than MCF-7 cell cultures, consistent with analysis of annexin V-7-AAD stained cells indicating that MDA-MB-231 possess anti-apoptotic ability 1.7 fold higher than MCF-7 cells. GeLC-MS/MS analysis of protein lysates of MDA-MB-231 and MCF-7 cells grown under both culture conditions identified 925 proteins which are differentially expressed, 54 of which were expressed only in suspended and adherent MDA-MB-231 but not in MCF-7 cells. These proteins have been implicated in various cellular processes, including DNA replication and repair, transcription, translation, protein modification, cytoskeleton, transport and cell signaling. Analysis based on the STITCH database predicted the interaction of phospholipases, PLC and PLD, and 14-3-3 beta/alpha, YWHAB, with the intrinsic and extrinsic apoptotic signaling network, suggesting putative roles in controlling anti-anoikis ability. MDA-MB-231 cells grown in the presence of inhibitors of phospholipase C, U73122, and phospholipase D, FIPI, demonstrated reduced ability to survive in suspension culture, indicating functional roles of PLC and PLD in the process of anti-anoikis. Our study identified intracellular mediators potentially associated with establishment of anoikis resistance of metastatic cells. These proteins require further clarification as prognostic and therapeutic targets for advanced breast cancer.

A Study on Experimental Characteristics in Fire Investigation Techniques of Flammable Liquids (유류화재의 감식기법의 실험적 특성에 관한 연구)

  • Hwang, Taeyeon;Choi, Donmook
    • Fire Science and Engineering
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    • v.26 no.6
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    • pp.7-14
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    • 2012
  • This paper is to develop analytical techniques of flammable liquids which have been used for accelerating fire in accidental fires and arsons. We tested the temperature distribution of ceiling, fire patterns on the floor, and existence of flammable liquids and a check with GC/MS about flammable liquids comparing with papers, newspapers, and clothing. Research findings are as follows. The temperature of ceiling is influenced by flame. So gasoline and thinner was observed that combustible materials would be burned by flame. The fire patten on the floor was observed that flammable liquids had specialized pattern comparing combustible materials. When combustible materials on the PVC (Polyvinyl chloride) floor was burned, they didn't react to the gas detector. But flammable liquids had opposite results. After 7 days, we identified components of fire residues with the GC/MS (Gas Chromatography/Mass Spectrometry) about existence of flammable liquids and got components of flammable liquids. Fire investigation is a complicated processes. But we understand characteristics of materials, need detail investigations, and use the GC/MS to analyse flammable materials.

Pharmacoeconomics Evaluation of Morphine, MS Contin and Oxycodone in the Treatment of Cancer Pain

  • Zhang, Wen-Zhou;Yu, Wei-Jiang;Zhao, Xiu-Li;He, Bao-Xia
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8797-8800
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    • 2014
  • Objective: To analyze cost-effectiveness of morphine, MS contin and oxycodone in the treatment of cancer pain, providing guidance for rational drug use in the clinic. Methods: Confirmed by histology, a total of 171 patients with various cancers who required analgesic treatment were selected and divided into 3 groups, 57 cases for each group, given morphine, MS contin and oxycodone, respectively. If there appeared a poor short-term effect or aggravated sudden pain during the treatment, a short-acting morphine injection was given and adverse reactions were processed by symptomatic treatment. The pain relief rate and adverse reactions of groups were observed and pharmacoeconomics evaluation was undertaken. Results: The pain relief rates with morphine, MS contin and oxycodone were 89.5%(51/57), 91.2%(52/57) and 93.0%(53/57), respectively, with no difference samong groups (${\chi}^2=4.4489$, P=0.6162). The occurrence rates of adverse reactions were 59.7%(34/57), 54.4%(31/57) and 43.9%(25/57), again with no significant variation (P>0.05). The ratios of cost-effectiveness (C/E) for the 3 groups were $14.6{\pm}7.21$, $15.0{\pm}7.44$ and $16.1{\pm}8.10$. When the price of 3 kinds of analgesics was reduced by 10%, the ratios of cost-effectiveness were $12.2{\pm}6.53$, ($13.4{\pm}6.08$ and $14.5{\pm}6.74$ but there was no differences when compared with before the price adjustment (t=1.86, P=0.0651; t=1.30, P=0.1948; t=1.17, P=0.2453). Conclusion: Morphine, MS contin and oxycodone give similar pain relief and adverse reaction rates but of all, morphine is the preferred drug for the treatment of cancer pain from the perspective of pharmacoeconomics.

Determination of Adrenosterone and its Metabolites in Human Urine by LC/APCI/MS and GC/MS

  • Han, Eun-Jung;Yim, Ok-Kyoung;Beak, Sun-Young;Chung, Jae-Yeon;Lee, Ji-Hye;Kim, Jun-Gahn;Kim, Yun-Je
    • Bulletin of the Korean Chemical Society
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    • v.30 no.7
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    • pp.1489-1496
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    • 2009
  • This study was done for the determination and excretion profile of adrenosterone and its metabolites in human urine using both liquid chromatography with atmospheric pressure chemical ionization mass spectrometry and gas chromatography with mass spectrometry. Adrenosterone and its two metabolites were detected in human urine after administration a healthy volunteer with 75 mg of adrenosterone. We found that adrenosterone-M1 ($C_{19}H_{26}O_3$) was a reduction and adrenosterone-M2 ($C_{19}H_{26}O_4$) was a hydroxylation at C-ring, which did not know the exact position of the C-ring. The adrenosterone parent was detected by GC/TOF-MS, but not detected by LC/APCI/MS because of low intensity. Adrenosterone and its two metabolites were excreted as their glucuronided fractions. The recovery of this method ranged from 100.7 to 118.4% and the reproducibility and accuracy test were 85.5 to 112.0% and 1.1 to 8.4%, respectively. The excretion studies showed that adrenosterone and its metabolites were detectable in human urine during a 48 h period after oral administration, with maximum level of excretion at 4.1 h. The glucuro-/sulfaconjugated ratio of adrenosterone, M1 and M2 was 0.73 ${\pm}$ 0.03, 0.96 ${\pm}$ 0.06 and 0.89 ${\pm}$ 0.03 (n = 6), respectively. The amounts of adrenosterone excreted in urine were 14.75 ng for 48 h. Also, the maximum level of androsterone and 11$\beta$-hydroxy androsterone, which were endogenous steroids, were reached 4.1 h after the oral administration of adrenosterone.

Targeted chiral lipidomics analysis of bioactive eicosanoid lipids in cellular systems

  • Lee, Seon-Hwa;Blair, Ian A.
    • BMB Reports
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    • v.42 no.7
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    • pp.401-410
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    • 2009
  • We have developed a targeted lipidomics approach that makes it possible to directly analyze chiral eicosanoid lipids generated in cellular systems. The eicosanoids, including prostaglandins (PGs), thromboxanes (TXs), leukotrienes (LTs) and alcohols (HETEs), have been implicated as potent lipid mediators of various biological processes. Enzymatic formations of eicosanoids are regioselective and enantioselective, whereas reactive oxygen species (ROS)-mediated formation proceeds with no stereo-selectivity. To distinguish between enzymatic and non-enzymatic pathways of eicosanoid formation, it is necessary to resolve enantiomeric forms as well as regioisomers. High sensitivity is also required to analyze the eicosanoid lipids that are usually present as trace amounts (pM level) in biological fluids. A discovery of liquid chromatography-electron capture atmospheric pressure chemical ionization/mass spectrometry (LC-ECAPCI/MS) allows us to couple normal phase chiral chromatography without loss of sensitivity. Analytical specificity was obtained by the use of collision-induced dissociation (CID) and tandem MS (MS/MS). With combination of stable isotope dilution methodology, complex mixtures of regioisomeric and enantiomeric eicosanoids have been resolved and quantified in biological samples with high sensitivity and specificity. Targeted chiral lipidomics profiles of bioactive eicosanoid lipids obtained from various cell systems and their biological implications have been discussed.

Analysis of Gingerol Compounds of Raw Ginger (Zingiber officinale Roscoe) and Its Paste by High Performance Liquid Chromatograph-Mass Spectrometry (LC/MS) (LC/MS에 의한 원료생강 및 생강 페이스트 중의 Gingerol 화합물 분석)

  • 조길석
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.5
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    • pp.747-751
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    • 2000
  • This study was carried out to improve the analysis method of gingerol compounds from ginger (Zingiber officinale Roscoe). Pungent components of ginger were extracted by acetone and lisolated by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) with LiChrosorb RP-18 column. Three homologues of gingerols were identified by HPLC-mass spectrometry (LC/MS) and nuclear magnetic resonance (NMR). The contents of [6]-, [8]- and [10]-gingerols in three homologues identified were 635.3 mg%, 206.6 mg% and 145.7 mg% in raw ginger, and were 418.2 mg%, 142.6 mg% and 103.3 mg% in ginger paste, respectively.

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Strength and Durability Properties of Concrete with Starch Admixture

  • Akindahunsi, A.A.;Uzoegbo, H.C.
    • International Journal of Concrete Structures and Materials
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    • v.9 no.3
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    • pp.323-335
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    • 2015
  • This paper examines some properties of concrete, such as strength, oxygen permeability and sorptivity using starch [cassava (CA) and maize (MS)] as admixtures. Concrete cubes containing different percentages of the CA and MS by weight of cement (0, 0.5, 1.0, 1.5 and 2.0 %) were cast. Compressive strength tests were carried out after 3, 7, 14, 21, 28, 56, 90, 180, 270 and 365 days of curing. Oxygen permeability and sorptivity tests were carried out on another set of concrete specimens with the same percentages of starch at 7, 28, 90, 180, 270 and 365 days. Oxygen permeability and sorptivity tests data obtained were subjected to Kruskal-Wallis one-way analysis of variance by ranks. The strength increase after 1 year over the control for CA 0.5 and CA 1.0 are 2.7 and 3.8 % respectively, while MS 0.5 and MS 1.0 gave 1.5 % increase over control. These results showed a decrease in oxygen permeability and rates of sorptivity, with concretes containing starch as admixtures giving better performance than the control concretes.

Identification for Flavones in Different Parts of Cirsium japonicum

  • Kim, Su-Jeong;Kim, Gun-Hee
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.330-335
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    • 2003
  • Cirsium japonicum is a herbaceous perennial plant grown worldwide, which has been used as a folklore medicine due to its anti-inflammatory properties. A few studies have reported its functional properties, but analytical methods that more confidently and reproductively analyze the flavonoids are required. To establish analytical methods for the detection of flavones in Cirsium japonicum, the potential of HPLC and LC/MS were investigated. For this, the plants were separated into 4 parts; the root, stem, leaves, and flowers. The flavones in each part of the dried materials were analyzed by HPLC. Identification of flavones was performed by LC/MS. The leaves and flowers of Cirsium japonicum gave the optimum peaks, which were not detected by HPLC in the other parts of plants. Using LC/MS, three kinds of flavones were tentatively identified from the leaves, which were thought to be luteolin (5,7,3',4'-tetrahydroxy-flavone), apigenin (4',5,7-trihy-droxyflavone), and hispidulin (4',5,7-trihydroxy-6-methoxyflavone). Two flavones were detected from the flowers, which were been assumed to be apigenin and luteolin.

Mornitoring of non-steroidal anti-inflammatory drugs in livestock products (축산식품 중 비스테로이드성 항염증 약물의 잔류함량 모니터링)

  • Choi, Yoon-Hwa;Kim, Yoen-Joo;Shin, Bang-Woo;Lee, Jung-Hark
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.285-289
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    • 2011
  • This study was conducted to determine the content of non-steroidal anti-inflammatory drugs (NSAIDs) in meats available on the Korean markets. The analysis was carried out using following conditions; C18 column ($100{\times}2.1$ mm, 1.7 ${\mu}m$), mobile phase composed of DW (containing 0.1% formic acid): acetonitrile (containing 0.1% formic acid), binary pump at a flow rate of 0.3 ml/min and 5 ${\mu}l$ of injection volume, MS/MS detector with ESI positive mode. The calibration range of five NSAIDs showed linearity ($r^2{\geq}0.99$) at concentration range of 3.125~200 ${\mu}g$/kg. The recoveries in fortified muscle more than 78.7~100.3%. The detection limits for meloxicam, ketoprofen, flunixin, carprofen and tolfenamic acid were 3.5, 1.6, 1.7, 9.8 and 4.8 ${\mu}g$/kg, respectively. We also monitored NSAIDs residue in cattle muscle 51 samples. The test results, NSAIDs were all not founded.

Quantification of Entacapone in Human Plasma by HPLC Coupled to ESI-MS/MS Detection: Application to Bioequavalence Study (체외에서 ESI-MS/MS 탐지에 연결된 HPLC에 의한 Entacapon의 수량화: 생물학적 동등성 연구에 적용)

  • Balasekhara Reddy., Ch.;Baburao., Ch.;Chandrasekhar., K.B.;Kanchanamala., K.;RihanaParveen., S.K.;Ravikumar., Konda
    • Journal of the Korean Chemical Society
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    • v.54 no.5
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    • pp.523-532
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    • 2010
  • The proposed method is simple, sensitive and specific Liquid chromatography-tandem mass spectrometry (LCESI-MS/MS) method for the quantification of Entacapone (EA) in human plasma using Entacapone-d10 (EAD10) as an internal standard (IS). Chromatographic separation was performed on Zorbax SB-C18, $2.1{\times}50\;mm$, $5\;{\mu}m$ column, mobile phase composed of 10 mM Ammonium formate (pH 3.0): Acetonitrile (60:40 v/v), with a flow-rate of 0.7 mL/min, followed by Liquid-liquid extraction. EA and EAD10 were detected with proton adducts at m/z $306.1{\rightarrow}233.1$ and $316.3{\rightarrow}233.0$ in multiple reaction monitoring (MRM) positive mode respectively. The method was validated over a linear concentration range of 1.00 - 2000.00 ng/mL with correlation coefficient ($r^2$) $\geq$ 0.9993. Intra and inter-day Precision within 3.60 to 7.30 and 4.20 to 5.50% and Accuracy within 97.30 to 104.20 and 98.30 to 105.80% proved for EA. This method is successfully applied in the bioequivalence study of healthy Indian human volunteers.