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MALDI matrix

Daniel Goldenberger, Vladimira Hinić, Spasenija Savic Prince, Michael Tamm, Anna-Maria Balestra, Doris Hohler, Reno Frei
BACKGROUND: Trichosporon mycotoxinivorans is a recently described yeast-like fungal organism and its association as a pathogen for patients with cystic fibrosis (CF) was reported previously. We show the clinical course of a CF patient over 9 years as well as the applications of modern molecular and proteomic identification techniques of this rare fungus. CASE PRESENTATION: We present the case of a 32-year-old male CF patient with sputum cultures continuously positive with the anamorphic yeast T...
October 25, 2016: BMC Infectious Diseases
Caiqiao Xiong, Xiaoyu Zhou, Qing He, Xi Huang, Jiyun Wang, Wen-Ping Peng, Huan-Cheng Chang, Zongxiu Nie
Mass is a fundamental physical property of individual cell, from which reveals the cell growth, cycle, and activity. Taking advantage of the cell mass spectrometry (CMS), accurate mass measurement of charged single cell has been achieved. However, with the increasing demand for high-efficiency single-cell analysis in biology, the limited throughput and inefficient cell desorption/ionization of the CMS inevitably becomes an important issue. To address the challenge, a state of the art visible-wavelength matrix assisted laser desorption/ionization (MALDI) CMS was developed...
October 26, 2016: Analytical Chemistry
Eun Ji Park, Dong Hee Na
The separation mechanism of PEGylated peptides in reversed-phase high-performance liquid chromatography (RP-HPLC) is complex, because the PEGylated molecules exhibit physicochemical properties that are different from those of the parent molecules and have heterogeneous structure. Since most separation studies have focused on the hydrophobicity of the peptide relative to poly(ethylene glycol) (PEG), the role of PEG in the separation of PEGylated peptides on RP-HPLC is not clear. To elucidate the effect of the attached PEG on the retention of PEGylated peptides on RP-HPLC, the mono-PEGylated forms of collagen pentapeptide and octreotide were fractionated drop-by-drop from the outlet of the HPLC system and each drop was subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)...
October 25, 2016: Analytical Chemistry
Eugenia Bertona, Adriana N De Paulis, Miguel A Gutiérrez, Victoria Santa María, Carlos A Vay, Silvia C Predari
Dermabacter hominis species is constituted by Gram-positive facultative anaerobic coryneform rods being part of the resident microbiota human skin, and exceptionally associated to infections in immunocompromised or severely debilitated patients. An immunocompetent young adult woman with a neck sebaceous cyst infected by D. hominis as unique etiologic agent is presented. Phenotypic identification of the causative agent was achieved through simple tests, based on the originally scheme proposed by Funke and Bernard, and feasible to be performed in a hospital Microbiology Laboratory...
October 20, 2016: Revista Argentina de Microbiología
Christopher D Doern, Susan M Butler-Wu
The performance of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MS) for routine bacterial and yeast identification as well as direct-from-blood culture bottle identification has been thoroughly evaluated in the peer-reviewed literature. Microbiologists are now moving beyond these methods to apply MS to other areas of the diagnostic process. This review discusses the emergence of advanced matrix-assisted laser desorption ionization time-of-flight MS applications, including the identification of filamentous fungi and mycobacteria and the current and future state of antimicrobial resistance testing...
October 15, 2016: Journal of Molecular Diagnostics: JMD
Christopher D Doern, Susan M Butler-Wu
The performance of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MS) for routine bacterial and yeast identification as well as direct-from-blood culture bottle identification has been thoroughly evaluated in the peer-reviewed literature. Microbiologists are now moving beyond these methods to apply MS to other areas of the diagnostic process. This review discusses the emergence of advanced matrix-assisted laser desorption ionization time-of-flight MS applications, including the identification of filamentous fungi and mycobacteria and the current and future state of antimicrobial resistance testing...
November 2016: Journal of Molecular Diagnostics: JMD
Wenjie Gao, Henghui Li, Yanyan Liu, Yuhong Liu, Xiaojun Feng, Bi-Feng Liu, Xin Liu
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has become an important technology for glycan analysis due to its ease of operation, short analysis time and impurity tolerance. However, the low ionization efficiency of N-glycans led to the difficulty in analyzing glycans of low abundance in complex biological samples due to the lack of basic site for protonation. Therefore, highly sensitive method for the glycans analysis is in urgent demand. Here we report a new strategy to introduce a permanent charge at the reducing end of N-linked glycans by a one pot reaction, where glycosylamines that were obtained by rapid deglycosylation within 5min were labeled with N-succinimidyloxycarbonylmethyl tris (2,4,6- trimethoxyphenyl) phosphonium bromide (TMPP-Ac-OSu)...
December 1, 2016: Talanta
Neha Kumari, Rajesh Kumar, Vandana Mishra, Savita Yadav
CC-NBS-LRR (CNL) plant proteins are related with highly conserved family of disease resistance protein distinguished by a coiled-coil domain, which plays an important role in innate immunity. The present study reports the purification and identification of CNL like protein fragment (CNL-LPF) by two step chromatography and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF/MS), respectively. Furthermore, current study also illustrated the development of polyclonal antibody against purified CNL-LPF, which was used for immunolocalization of CNL-LPF in cytoplasm of cotyledon, using Fluorescence microscopy and Transmission electron microscopy...
October 20, 2016: Protein Journal
Caiyun He, Guori Gao, Jianguo Zhang, Aiguo Duan, Hongmei Luo
BACKGROUND: Low temperature is one of the crucial environmental factors limiting the productivity and distribution of plants. Sea buckthorn (Hippophae rhamnoides L.), a well recognized multipurpose plant species, live successfully in in cold desert regions. But their molecular mechanisms underlying cold tolerance are not well understood. METHODS: Physiological and biochemical responses to low-temperature stress were studied in seedlings of sea buckthorn. Differentially expressed protein spots were analyzed using multiplexing fluorescent two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption/ionization (MALDI) time-of-flight/time-of-flight (TOF/TOF) mass spectrometry (MS), the concentration of phytohormone was measured using enzyme-linked immunosorbent assay, and a spectrophotometric assay was used to measure enzymatic reactions...
2016: Proteome Science
Katia Roberta A Belaz, Alessandra Tata, Moana R França, Maressa I Santos da Silva, Pedro Henrique Vendramini, Anna Maria A P Fernandes, Fábio L D'Alexandri, Marcos N Eberlin, Mario Binelli
Phospholipid metabolism and signaling influences on early pregnancy events in cattle are unknown. This study aimed to characterize global phospholipid composition of oviduct and uterus during early diestrus in a model of contrasting embryo receptivity. Beef cows were treated to ovulate a larger (LF-LCL group, associated with greater receptivity) or smaller follicle (SF-SCL group) and, consequently, to present greater or smaller plasma concentrations of estradiol during proestrus-estrus as well as progesterone during early diestrus...
October 19, 2016: Biology of Reproduction
T Banach, M Bochniarz, P Łyp, Ł Adaszek, W Wawron, B Furmaga, M Skrzypczak, J Ziętek, S Winiarczyk
The aim of this study was to use matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of coagulase-negative staphylococci (CNS) isolated from the milk of cows with subclinical mastitis. The study material consisted of 33 isolates of CNS, identified by the results of API Staph tests, obtained from the milk of cows with subclinical mastitis. Based on the spectra analyses, MALDI-TOF MS tests of 33 bacterial samples allowed identification of the microorganisms in 27 cases (81...
September 1, 2016: Polish Journal of Veterinary Sciences
Rima Ait-Belkacem, Marialaura Dilillo, Davide Pellegrini, Avinash Yadav, Erik L de Graaf, Liam A McDonnell
Atmospheric pressure MALDI on a Q-Exactive instrument was optimized for in-source decay and pseudo-MS(3). The dependence of AP-MALDI ISD on the MALDI liquid matrix was investigated for peptides and proteins. The liquid matrices enabled long-life ISD signal, and exhibited high fragment ion yield and signal stability. Extensive a-, b-, c-, y-, and z-type fragment series were observed depending on the matrix used but were most extensive with 2,5-DHB. Complete sequence coverage of small peptide and intact protein-terminus sequence tags were obtained and confirmed using HCD as a pseudo-MS(3) method...
October 17, 2016: Journal of the American Society for Mass Spectrometry
Jun Hwa Lee, Byong Chul Yoo, Yun Hwan Kim, Sun-A Ahn, Seung-Gu Yeo, Jae Youl Cho, Kyung-Hee Kim, Seung Cheol Kim
BACKGROUND: A low-mass-ion discriminant equation (LOME) was constructed to investigate whether systematic low-mass-ion (LMI) profiling could be applied to ovarian cancer (OVC) screening. RESULTS: Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry was performed to obtain mass spectral data on metabolites detected as LMIs up to a mass-to-charge ratio (m/z) of 2500 for 1184 serum samples collected from healthy individuals and patients with OVC, other types of cancer, or several types of benign tumor...
2016: BioData Mining
Syed Ghulam Musharraf, Mariam Ameer, Arslan Ali
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) being soft ionization technique, has become a method of choice for high-throughput analysis of proteins and peptides. In this study, we have explored the potential of atypical anti-psychotic drug olanzapine (OLZ) as a matrix for MALDI-MS analysis of peptides aided with the theoretical studies. Seven small peptides were employed as target analytes to check performance of olanzapine and compared with conventional MALDI matrix α-cyano-4-hydroxycinnamic acid (HCCA)...
October 5, 2016: Journal of Pharmaceutical and Biomedical Analysis
Guangming Gong, Wenya Liu, Shudong Wang
This study aimed to reduce the cytotoxicity and improve the targeting of aclacinomycin (ACM) by covalently coupling it with amino-oxyacetic acid (AOA) to generate an active intermediate, AOA-ACM. AOA-ACM was conjugated with self-assembled human serum albumin (HSA) nanoparticles constructed using tris(2-carboxyethyl)phosphine (TCEP) as disulfide bond breaking molecules in an 'opening stage-intermediate-closing stage' route, in which the hydrophobic interaction, interchange of sulfhydryl and hydrogen bond may be the key factors in the assembling process...
October 17, 2016: Nanotechnology
Xin Zhou, Ping Wang, Ying-Jian Zhang, Jin-Jin Xu, La-Mei Zhang, Lin Zhu, Li-Ping Xu, Xiao-Min Liu, Huan-Huan Su
The present study aimed to investigate the proteomic difference between melanosis coli (MC) alone and melanosis coli with colon cancer (MCCC). Protein expression in patients with different diseases was analyzed using two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/TOF-MS). A total of 14 protein differences with a confidence level of >95% were found. There were six differences between MC and normal tissues, in which two proteins exhibited upregulated expression levels and four proteins exhibited downregulated expression levels in MC...
October 17, 2016: Oncology Reports
Hai-Long Liu, Yu-Jie Chang, Ting Fan, Zhi-Yuan Gu
Stable 2-D metal-organic framework nanosheets were utilized as a superior clean-background matrix for MALDI-TOF MS analysis of small biomolecules and pollutants in both positive and negative ion modes. The matrix could unusually afford up to 1000 mM of the salt concentrations in the monitoring of the enzymatic hydrolysis of neurotransmitter acetylcholine.
October 17, 2016: Chemical Communications: Chem Comm
Didrik Hjertaker Grevskott, Cecilie Smith Svanevik, Astrid Louise Wester, Bjørn Tore Lunestad
Continuous European Union programmes with specified methods for enumeration of Escherichia coli in bivalves for human consumption are currently running. The objective of this research was to examine the species accuracy of the five times three tube Most Probable Number (MPN) EU reference method used for detection of E. coli in marine bivalves. Among 549 samples of bivalves harvested from Norwegian localities during 2014 and 2015, a total number of 200 bacterial isolates were prepared from randomly selected culture-positive bivalves...
October 13, 2016: Journal of Microbiological Methods
M Mulet, M Gomila, A Ramírez, S Cardew, E R B Moore, J Lalucat, E García-Valdés
Fifty-two Pseudomonas strains that were difficult to identify at the species level in the phenotypic routine characterizations employed by clinical microbiology laboratories were selected for genotypic-based analysis. Species level identifications were done initially by partial sequencing of the DNA dependent RNA polymerase sub-unit D gene (rpoD). Two other gene sequences, for the small sub-unit ribosonal RNA (16S rRNA) and for DNA gyrase sub-unit B (gyrB) were added in a multilocus sequence analysis (MLSA) study to confirm the species identifications...
October 15, 2016: European Journal of Clinical Microbiology & Infectious Diseases
Alba Hykollari, Katharina Paschinger, Barbara Eckmair, Iain B H Wilson
N-glycans from invertebrates and protists have often unusual structures which present analytical challenges. Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps...
2017: Methods in Molecular Biology
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