ABSTRACT
Due to the excellent advantages of high speed, high precision, and driving force, piezoelectric actuators nanopositioning systems have been widely used in various micro/nanomachining fields. However, the inherent resonance dynamic of the nanopositioning system generated by the flexure-hinge greatly deteriorates the positioning performance and limits the closed-loop bandwidth. Even worse, the notch filter for eliminating the effect of resonance does not work due to the varying resonant frequency resulting from the external disturbance or mass load. To this end, an adaptive notch filter for piezo-actuated nanopositioning system via position and online estimate dual-mode (POEDM) has been proposed in this paper, which can estimate the varying resonant frequency in real-time and suppress the resonance to improve the closed-loop bandwidth. First, a novel variable forgetting factor recursive least squares (VFF-RLS) algorithm for estimating resonant frequency online is presented, which is robust to the noise and provides the performances of both fast tracking and stability. Then, a POEDM method is proposed to achieve the online identification of the resonant frequency in the presence of noise and disturbance. Finally, a series of validation simulations are carried out, and the results indicate that, the frequency of input signal and the bandwidth have been achieved up to 12.5% and 87.5% of the first resonant frequency, respectively.
ABSTRACT
Breast cancer is the second leading cause of cancer-related deaths in women. The need for new clinical biomarkers in breast cancer is necessary to further predict prognosis and therapeutic response. In this article, the LC-MS histone H1 phosphorylation profiles were established for three distinct breast cancer cell lines. The results show that the extent of H1 phosphorylation can distinguish between the different cell lines. The histone H1 from the metastatic cell line, MDA-MB-231, was subjected to chemical derivitization and LC-MS/MS analysis. The results suggest that the phosphorylation at threonine 146 is found on both histone H1.2 and histone H1.4. Cell lines were then treated with an extracellular stimulus, estradiol or kinase inhibitor LY294002, to monitor changes in histone H1 phosphorylation. The data show that histone H1 phosphorylation can increase and decrease in response to extracellular stimuli. Finally, primary breast tissues were stained for the histone H1 phosphorylation at threonine 146. Variable staining patterns across tumor grades and subtypes were observed with pT146 labeling correlating with tumor grade. These results establish the potential for histone H1 phosphorylation at threonine 146 as a clinical biomarker in breast cancer.
Subject(s)
Breast Neoplasms/metabolism , Histones/metabolism , Phosphoproteins/metabolism , Amino Acid Sequence , Blotting, Western , Breast Neoplasms/pathology , Cell Line , Cell Line, Tumor , Chromatography, Liquid , Chromones/pharmacology , Enzyme Inhibitors/pharmacology , Estradiol/pharmacology , Female , Humans , Immunohistochemistry , MCF-7 Cells , Mass Spectrometry/methods , Molecular Sequence Data , Morpholines/pharmacology , Phosphatidylinositol 3-Kinases/metabolism , Phosphoinositide-3 Kinase Inhibitors , Phosphorylation/drug effects , Protein Isoforms/metabolism , Proteomics/methods , Threonine/metabolismABSTRACT
Paper spray (PS) ionization, an ambient ionization method, has previously been explored as a direct and fast method for mass spectrometric analysis of complex mixtures. It has been applied to the analysis of a wide variety of compounds, mostly small molecules. The work reported here extends the application of PS ionization to noncovalent protein complexes on an ion mobility tandem mass spectrometer. Similar mass spectra for protein complexes were obtained by PS ionization and nanoflow electrospray ionization (nESI), indicating that intact protein complexes can be preserved in PS ionization. In addition, collisional cross sections measured by ion mobility provide evidence that the protein assemblies may remain compact by PS ionization. With PS, it is possible to detect hemoglobin tetramer from a blood sample with minimal sample preparation. This is the first report to show that PS ionization is a promising ionization method for nonconvalent protein complexes.
Subject(s)
Blood Proteins/chemistry , Mass Spectrometry/methods , Paper , Humans , Models, Molecular , Protein ConformationABSTRACT
A custom in-line surface-induced dissociation (SID) device has been incorporated into a commercial ion mobility quadrupole/time-of-flight mass spectrometer in order to provide an alternative and potentially more informative activation method than the commonly used collision-induced dissociation (CID). Complicated sample mixtures can be fractionated by ion mobility (IM) and then dissociated by CID or SID for further structural analysis. Interpretation of SID spectra for cesium iodide clusters was greatly simplified with IM prior to dissociation because products originating from different precursors and overlapping in m/z but separated in drift time can be examined individually. Multiple conformations of two protein complexes, source-activated transthyretin tetramer and nativelike serum amyloid P decamer, were separated in ion mobility and subjected to CID and SID. CID spectra of the mobility separated conformations are similar. However, drastic differences can be observed for SID spectra of different conformations, implying different structures in the gas phase. This work highlights the potential of utilizing IM-SID to study quaternary structures of protein complexes and provides information that is complementary to our recently reported SID-IM approach.
