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871.
Ling-Zhi Wang Daniel Chan Winnie Yeo Seow-Ching Wan Stephen Chan Anthony Chan Soo-Chin Lee How-Sung Lee Boon-Cher Goh 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(26):2409-2414
A novel, sensitive and reliable liquid chromatography–tandem mass spectrometric (LC–MS/MS) method was developed and validated for the determination of belinostat (PXD101) in human plasma. Oxamflatin was used as the internal standard. Liquid–liquid extraction of the plasma sample was performed using tert-butyl methyl ether as the organic solvent. Chromatographic separation was achieved on a BDS Hypersil C18 column (2.1 mm × 100 mm, 5 μm) using gradient elution mode using 0.05% formic acid in water and 0.05% formic acid in acetonitrile as solvents A and B, respectively, 60/40. The run time was 6 min. The mass spectrometer was operated under a positive electrospray ionization condition and a multiple reaction monitoring mode. An excellent linear calibration was achieved in the range of 0.5–1000 ng/mL. An average recovery of belinostat for four quality controls was 72.6% and the recovery of the internal standard at 1000 ng/mL was 67.8%. The intra-day and inter-day precisions for belinostat were ≤8.0 and ≤10.3%, respectively, and their accuracy ranged from 100.2 to 106.7%. No significant matrix effect was identified. In analysis of patient samples, belinostat glucuronide was identified and baseline separated from belinostat. This well-validated assay has been applied for quantification of belinostat in plasma samples within 24 h after the start of infusion for Asian hepatocellular carcinoma patients in a dose escalation study. 相似文献
872.
Tamiika K. Hurst Da Wang Richard B. Thompson Carol A. Fierke 《Biochimica et Biophysica Acta - Proteins and Proteomics》2010,1804(2):393-403
Carbonic anhydrases are archetypical zinc metalloenzymes and as such, they have been developed as the recognition element of a family of fluorescent indicators (sensors) to detect metal ions, particularly Zn2+ and Cu2+. Subtle modification of the structure of human carbonic anhydrase II isozyme (CAII) alters the selectivity, sensitivity, and response time for these sensors. Sensors using CAII variants coupled with zinc-dependent fluorescent ligands demonstrate picomolar sensitivity, unmatched selectivity, ratiometric fluorescence signal, and near diffusion-controlled response times. Recently, these sensors have been applied to measuring the readily exchangeable concentrations of zinc in the cytosol and nucleus of mammalian tissue culture cells and concentrations of free Cu2+ in seawater. 相似文献
873.
Sophia Letsiou Ying Lu Tzortzis Nomikos Smaragdi Antonopoulou Demosthenes Panagiotakos Christos Pitsavos Christodoulos Stefanadis Spiros A. Pergantis 《Proteomics》2010,10(19):3447-3457
In this study, a method, based on dual column affinity chromatography hyphenated to isotope dilution inductively coupled plasma–quadrupole MS, was developed for selenium determination in selenoprotein P, glutathione peroxidase, and selenoalbumin in human serum samples from a group of healthy volunteers (n=399). Method improvement was achieved using methanol‐enhanced isotope dilution which resulted in improved sensitivity and removal of isobaric interferences. Although no human serum reference materials are currently certified for their selenium species levels, method development was conducted using human serum reference material BCR 637 and 639 as their Se species content has been reported in the previous studies, and thus comparisons were possible. The mean selenium concentrations determined for the 399 healthy volunteer serum samples were 23±10 ng Se mL?1 for glutathione peroxidase, 49±15 ng Se mL?1 for selenoprotein P and 11±4 ng Se mL?1 for selenoalbumin. These values are found to be in close agreement with published values for a limited number of healthy volunteer samples, and to establish baseline Se levels in serum proteins for an apparently healthy group of individuals, thus allowing for subsequent comparisons with respective values determined for groups of individuals with selenium related health issues, as well as assist in the discovery of potential selenium biomarkers. Also, the relationship between Se serum protein levels and some anthropometric characteristics of the volunteer population were investigated. Additionally, further development of the analytical method used in this study was achieved by adding a size exclusion chromatography column after the two affinity columns via a switching valve. This allowed for the separation of small selenium‐containing molecules from glutathione peroxidase and thus enhanced the overall confidence in its identification. 相似文献
874.
微小核糖核酸(microRNA,miRNA)在生物体的生长、发育及疾病的发生发展等过程中起着十分重要的作用。目前,人们关注的主要是miRNA在组织细胞内的功能。然而,最近的研究发现血清/血浆中存在一些循环的miRNA,血清/血浆miRNA在肿瘤、糖尿病等多种疾病中呈特异性表达,提示血清miRNA可作为潜在的生物标记物用于疾病的诊断和治疗。 相似文献
875.
渗透调节对冷敏感大豆[Glycine max(L.)Merr.]种子膜类脂组成和脂肪酸含量的影响 总被引:3,自引:0,他引:3
选用冷敏感的大豆[Gzycine max(L.)Merr.]品种“中黄22”为试材,运用薄层层析法和气相色谱法研究了渗控增强大豆种子活力过程中膜类脂的组成及变化规律,揭示生物膜类脂在抗吸胀冷害中的作用。结果表明,用33%(W/V)的PEG6000处理种子,随着渗控时间的延长,种子的发芽指数、活力指数和根干重都逐渐增加,在渗控72h时,大豆种子中不饱和磷脂如PC、PE的比例增加,各极性脂的脂肪酸组成也发生了变化,PC、PE和PI中饱和脂肪酸16:0的含量显著下降,而不饱和脂肪酸18:2明显上升,使它们的不饱和指数大大提高,并与渗控时间呈正相关。由以上结果可证明,经PEG渗控处理后,提高了冷敏感大豆膜的流动性,为防止吸胀冷害,保持膜系统的完整性和生物膜正常的代谢提供了前提条件。 相似文献
876.
877.
Rossi JP Villamil AM Echarte MM Alzugaray ME Borelli MI García ME Pande J Grover AK Gagliardino JJ 《Cell biochemistry and biophysics》2006,46(3):193-200
The aim of this study was to quantify the glucose modulation of the plasma membrane calcium pump (PMCA) function in rat pancreatic
islets. Ca2+-ATPase activity and levels of phosphorylated PMCA intermediates both transiently declined to a minimum in response to stimulation
by glucose. Strictly dependent on Ca2+ concentration, this inhibitory effect was fully expressed at physiological concentrations of the cation (less than 0.5 μM), then progressively diminished at higher concentrations. These results, together with those previously reported on the effects
of insulin secretagogues and blockers on the activity, expression and cellular distribution of the PMCA, support the concept
that the PMCA plays a key role in the regulation of Ca2+ signaling and insulin secretion in pancreatic islets. 相似文献
878.
Rawdkuen S Benjakul S Visessanguan W Lanier TC 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2006,144(4):544-552
A high-molecular-weight cysteine proteinase inhibitor (CPI) was purified from chicken (Gallus gallus) plasma using polyethylene glycol (PEG) fractionation and affinity chromatography on carboxymethyl–papain–Sepharose-4B. The CPI was purified 96.8-fold with a yield of 28.9%. Based on inhibitory activity staining for papain, CPI was shown to have an apparent molecular mass of 122 kDa. No inhibitory activity was obtained under reducing condition, indicating that CPI from chicken plasma was stabilized by disulfide bonds. CPI was stable in temperature ranges from 40 to 70 °C for 10 min; however, more than 50% of the inhibitory activity towards papain was lost within 30 min of heating at 90 °C. CPI was stable in the presence of salt up to 3%. The purified CPI exhibited the inhibitory activity toward autolysis of arrowtooth flounder (Atheresthes stomias) and Pacific whiting (Merluccius productus) natural actomyosin (NAM) in a concentration-dependent manner. 相似文献
879.
880.