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Ravirajsinh N Jadeja, Menaka C Thounaojam, Sandeep Khurana
In cirrhosis, changes in pressure-mediated vascular tone, a key determinant of systemic vascular resistance (SVR), are unknown. To address this gap in knowledge, we assessed ex vivo dynamics of pressurized mesenteric resistance arteries (diameter ~ 260 μm) from bile duct-ligated (BDL) and sham-operated (SHAM) rats and determined the underlying mechanisms. At isobaric intraluminal pressure (70 mmHg) as well as with step-wise increase in pressure (10-110 mmHg), arteries from SHAM-rats constricted more than BDL-rats, and had reduced luminal area...
February 16, 2017: Oncotarget
Lingxia Wang, Dezhuo Pan, Meng Liang, Yakubu Saddeeq Abubakar, Jian Li, Jinke Lin, Shipin Chen, Wei Chen
Plant anthocyanin biosynthesis is well understood, but the regulatory mechanism in purple foliage tea remains unclear. Using isobaric tag for relative and absolute quantification (iTRAQ), 815 differential proteins were identified in the leaves of Zijuan tea, among which 20 were associated with the regulation of anthocyanin metabolism. We found that the abundances of anthocyanin synthesis-related enzymes such as chalcone synthase, chalcone isomerase, dihydroflavonol 4-reductase and anthocyanin synthetase, as well as anthocyanin accumulation-related UDP-glucosyl transferase and ATP-binding cassette (ABC) transporters in the purple leaves were all significantly higher than those in the green leaves...
April 19, 2017: International Journal of Molecular Sciences
Christopher S Hughes, Victor Spicer, Oleg V Krokhin, Gregg B Morin
Methods for isobaric-tagged peptide analysis (e.g., TMT, iTRAQ), such as the synchronous precursor selection (SPS) tandem MS/MS/MS (MS(3)) approach, enable maintenance of reporter ion accuracy and precision by reducing the ratio compression caused by coisolated precursor ions. However, the decreased throughput of the MS(3) approach necessitates careful optimization of acquisition strategies and methods to ensure maximal proteome coverage. We present a systematic analysis of acquisition parameters used to analyze isobaric-tagged peptide samples on current generation Orbitrap mass spectrometer (MS) hardware...
April 20, 2017: Journal of Proteome Research
Christopher S Hughes, Chenchen Zhu, Victor Spicer, Oleg V Krokhin, Gregg B Morin
Multiplexed quantification with isobaric chemical tags (e.g., TMT, iTRAQ) provides a robust and efficient means to comparatively examine proteome dynamics between several biological states using a mass spectrometer (MS). The quantitative nature of isobaric tags necessitates strict validation of the observed ion signals in the chosen MS detector before differential patterns are extracted between biological states. We present an in-depth analysis of isobaric tag data acquired on current generation Orbitrap MS hardware to illustrate pitfalls in acquisition settings that can negatively impact results...
April 20, 2017: Journal of Proteome Research
Jingwen Xu, Yunlun Li, Shijun Zhang, Haiqiang Jiang, Nan Wang, Haiqing Lin
Tengfu Jiangya Tablet (TJT) is a well accepted antihypertension drug in China and its major active components were Uncaria total alkaloids and Semen Raphani soluble alkaloid. To further explore treatment effects mechanism of TJT on essential hypertension, a serum proteomic study was performed. Potential biomarkers were quantified in serum of hypertension individuals before and after taking TJT with isobaric tags for relative and absolute quantitation (iTRAQ) coupled two-dimensional liquid chromatography followed electrospray ionization-tandem mass spectrometry (2D LC-MS/MS) proteomics technique...
2017: Evidence-based Complementary and Alternative Medicine: ECAM
P Prikryl, L Vojtova, D Maixnerova, M Vokurka, M Neprasova, T Zima, V Tesar
Proteinuria is often used as a surrogate marker in monitoring and predicting outcome in patients with chronic kidney diseases, but it is non-specific. IgAN belongs to the most common primary glomerulonephritis worldwide with serious prognosis. The main aim of this work was to assess differences in urine proteins in patients with IgA nephropathy and to identify abnormal proteins as potential biomarkers of IgA nephropathy or the renal disease. In our pilot project, we selected 20 patients and compared them with 20 healthy volunteers...
April 12, 2017: Physiological Research
Shinya Matsuoka, Alissa Armstrong, Leesa L Sampson, Kaitlin M Laws, Daniela Drummond-Barbosa
Nutrients affect adult stem cells through complex mechanisms involving multiple organs. Adipocytes are highly sensitive to diet and have key metabolic roles, and obesity increases the risk for many cancers. How diet-regulated adipocyte metabolic pathways influence normal stem cell lineages, however, remains unclear. Drosophila melanogaster has highly conserved adipocyte metabolism and a well-characterized female germline stem cell (GSC) lineage response to diet. Here, we conducted an isobaric tags for relative and absolute quantification (iTRAQ) proteomic analysis to identify diet-regulated adipocyte metabolic pathways that control the female GSC lineage...
April 10, 2017: Genetics
Qing Yu, Xudong Shi, Yu Feng, K Craig Kent, Lingjun Li
Mass spectrometry (MS)-based isobaric labeling has undergone rapid development in recent years due to its capability for high throughput quantitation. Apart from its originally designed use with collision-induced dissociation (CID) and higher-energy collisional dissociation (HCD), isobaric tagging technique could also work with electron-transfer dissociation (ETD), which provides complementarity to CID and is preferred in sequencing peptides with post-translational modifications (PTMs). However, ETD suffers from long reaction time, reduced duty cycle and bias against peptides with lower charge states...
May 22, 2017: Analytica Chimica Acta
Lijun Zhang, Xiaofang Jia, Jun-O Jin, Hongzhou Lu, Zhimi Tan
Human immunodeficiency virus-1 (HIV-1) mainly relies on host factors to complete its life cycle. Hence, it is very important to identify HIV-regulated host proteins. Proteomics is an excellent technique for this purpose because of its high throughput and sensitivity. In this review, we summarized current technological advances in proteomics, including general isobaric tags for relative and absolute quantitation (iTRAQ) and stable isotope labeling by amino acids in cell culture (SILAC), as well as subcellular proteomics and investigation of posttranslational modifications...
April 5, 2017: Genomics, Proteomics & Bioinformatics
Shakir Shakir, Joelle Vinh, Giovanni Chiappetta
The redox conditions that reign inside a cell have a determining effect on a number of biological processes. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are key redox players and have been linked to a number of pathologies. They have also been shown to play an important regulating role in cell signaling events. On the proteome level, thiol groups of cysteinyl side chains constitute the major targets of ROS and RNS. A number of analytical techniques based on mass spectrometry have been developed to characterize the cysteine redoxome, often facing a number of technical challenges, mostly related to the lability, heterogeneity, and low abundance of the oxidized forms...
April 7, 2017: Analytical and Bioanalytical Chemistry
Nhu T N Phan, Marwa Munem, Andrew G Ewing, John S Fletcher
Lipids are abundant biomolecules performing central roles to maintain proper functioning of cells and biological bodies. Due to their highly complex composition, it is critical to obtain information of lipid structures in order to identify particular lipids which are relevant for a biological process or metabolic pathway under study. Among currently available molecular identification techniques, MS/MS in secondary ion mass spectrometry (SIMS) imaging has been of high interest in the bioanalytical community as it allows visualization of intact molecules in biological samples as well as elucidation of their chemical structures...
April 7, 2017: Analytical and Bioanalytical Chemistry
Jaeho Paek, Marian Kalocsay, Dean P Staus, Laura Wingler, Roberta Pascolutti, Joao A Paulo, Steven P Gygi, Andrew C Kruse
G-protein-coupled receptors (GPCRs) play critical roles in regulating physiological processes ranging from neurotransmission to cardiovascular function. Current methods for tracking GPCR signaling suffer from low throughput, modification or overexpression of effector proteins, and low temporal resolution. Here, we show that peroxidase-catalyzed proximity labeling can be combined with isobaric tagging and mass spectrometry to enable quantitative, time-resolved measurement of GPCR agonist response in living cells...
April 6, 2017: Cell
Michael von Domaros, Eva Perlt
The well-established quantum cluster equilibrium (QCE) model provides a statistical thermodynamic framework to apply high-level ab initio calculations of finite cluster structures to macroscopic liquid phases using the partition function. So far, the harmonic approximation has been applied throughout the calculations. In this article, we apply an important correction in the evaluation of the one-particle partition function and account for anharmonicity. Therefore, we implemented an analytical approximation to the Morse partition function and the derivatives of its logarithm with respect to temperature, which are required for the evaluation of thermodynamic quantities...
March 28, 2017: Journal of Chemical Physics
K J Lee, H Vanheel, T Vanuytsel, R Vos, J Tack
BACKGROUND: Altered duodenal sensorimotor responses to acid have been reported in a subset of patients with functional dyspepsia. To investigate whether NO is involved in these abnormalities, the effect of sildenafil (activates the NO pathway) on duodenal motor and sensory responses to acid in healthy humans was evaluated. METHODS: A barostat-manometry catheter including an infusion tube was positioned in the duodenum of 12 healthy volunteers. Duodenal motility and dyspeptic symptoms were evaluated during the whole study...
April 6, 2017: Neurogastroenterology and Motility: the Official Journal of the European Gastrointestinal Motility Society
Mariosimone Zoccali, Kevin A Schug, Phillip Walsh, Jonathan Smuts, Luigi Mondello
The present paper is focused on the use of a vacuum ultraviolet absorption spectrometer (VUV) for gas chromatography (GC), within the context of flow modulated comprehensive two-dimensional gas chromatography (FM GC×GC). The features of the VUV detector were evaluated through the analysis of petrochemical and fatty acids samples. Besides responding in a predictable fashion via Beer's law principles, the detector provides additional spectroscopic information for qualitative analysis. Virtually all chemical species absorb and have unique gas phase absorption features in the 120-240nm wavelength range monitored...
March 28, 2017: Journal of Chromatography. A
Dipy M Vasa, Peter L D Wildfong
Ribavirin (C8H12N4O5; anti-viral agent) was crystallized as two unique, phase-pure polymorphs (R-I and R-II). Calorimetrically determined isobaric heat capacities and heat of transition data were utilized to determine the solid-state transition temperature (Ttr), confirming enantiotropism, while R-I was determined to be kinetically stable at ambient temperature. Unprocessed samples of the low Tm polymorph, R-II, did not convert into R-I when held isothermally well above Ttr for 7days. In contrast milled R-II completely transformed to R-I after 15min at the same storage conditions, indicating that defects sustained during processing reduced the energy barrier for transformation, allowing it to occur...
April 2, 2017: International Journal of Pharmaceutics
M Dilillo, R Ait-Belkacem, C Esteve, D Pellegrini, S Nicolardi, M Costa, E Vannini, E L de Graaf, M Caleo, L A McDonnell
MALDI mass spectrometry imaging is able to simultaneously determine the spatial distribution of hundreds of molecules directly from tissue sections, without labeling and without prior knowledge. Ultra-high mass resolution measurements based on Fourier-transform mass spectrometry have been utilized to resolve isobaric lipids, metabolites and tryptic peptides. Here we demonstrate the potential of 15T MALDI-FTICR MSI for molecular pathology in a mouse model of high-grade glioma. The high mass accuracy and resolving power of high field FTICR MSI enabled tumor specific proteoforms, and tumor-specific proteins with overlapping and isobaric isotopic distributions to be clearly resolved...
April 4, 2017: Scientific Reports
Songbai Yang, Yue Pei, Ayong Zhao
Pseudorabies virus (PRV) is one of the most important pathogens of swine, resulting in severe economic losses to the pig industry. To improve our understanding of the host responses to PRV infection, we applied isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with liquid chromatography-tandem mass spectrometry to quantitatively identify the differentially expressed cellular proteins in PRV-infected PK15 cells. In total, relative quantitative data were identified for 4333 proteins in PRV and mock- infected PK15 cells, among which 466 cellular proteins were differentially expressed, including 234 upregulated proteins and 232 downregulated proteins...
April 4, 2017: Scientific Reports
Yonghoon Lee, Jose Chirinos, Jhanis Gonzalez, Dayana Oropeza, Vassilia Zorba, Xianglei Mao, Jonghyun Yoo, Richard E Russo
We evaluated the performance of laser ablation analysis techniques such as laser-induced breakdown spectroscopy (LIBS), laser ablation inductively coupled optical emission spectrometry (LA-ICP-OES), and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), in comparison with that of ICP-OES using aqueous solutions for the quantification of sulfur (S) in edible salts from different geographical origins. We found that the laser ablation based sampling techniques were not influenced by loss of S, which was observed in ICP-OES with aqueous solutions for a certain salt upon their dissolution in aqueous solutions, originating from the formation of volatile species and precipitates upon their dilution in water...
April 2017: Applied Spectroscopy
Zuchra Zakirova, Jon Reed, Gogce Crynen, Lauren Horne, Samira Hassan, Venkatarajan Mathura, Michael Mullan, Fiona Crawford, Ghania Ait-Ghezala
PURPOSE: Long term consequences of combined pyridostigmine bromide and permethrin exposure in C57BL6/J mice using a well characterized mouse model of exposure to these Gulf War (GW) agents were explored at the protein level. EXPERIMENTAL DESIGN: We used orthogonal proteomic approaches to identify pathways that are chronically impacted in the mouse CNS due to semi-acute GW agent exposure early in life. These analyses were performed on soluble and membrane-bound protein fractions from brain samples using two orthogonal isotopic labeling LC-MS/MS proteomic approaches - stable isotope dimethyl labeling and isobaric tags for relative and absolute quantitation...
March 28, 2017: Proteomics. Clinical Applications
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