291 THE USE OF THE DYNAMIC IMPACT APPROACH AND DESORPTION ELECTROSPRAY IONIZATION - MASS SPECTROSCOPY TO ANALYZE ADIPOGENESIS IN PORCINE ADIPOSE-DERIVED STEM CELLS

2013 ◽  
Vol 25 (1) ◽  
pp. 293 ◽  
Author(s):  
M. Bionaz ◽  
C. R. Ferreira ◽  
E. Monaco ◽  
R. G. Cooks ◽  
M. B. Wheeler
Keyword(s):  
Lipid Profile ◽  
Adipogenic Differentiation ◽  
Desorption Electrospray Ionization ◽  
Ionization Mass ◽  
Adipose Derived Stem Cells ◽  
Dynamic Impact ◽  
Electrospray Ionization Mass Spectroscopy ◽  
Ionization Mass Spectroscopy ◽  
Transcriptomics Data ◽  
Desi Ms

The measurement of end products of metabolism in combination with the dynamic measurement of the transcriptome can be a valuable approach to study complex processes, such as adipogenesis. For this purpose, we have combined the analysis of metabolic pathways using transcriptomics data with in situ lipid imaging obtained by desorption electrospray ionization–mass spectroscopy (DESI-MS). The transcriptome data was obtained from a previous experiment where adipose-derived stem cells (ASC) from 3 individual pigs were induced to differentiate into adipocytes for 21 days. The metabolic pathway analysis of transcriptomics data was performed using the dynamic impact approach (DIA) to determine the directional impact of pathways and gene categories. For the DESI-MS, porcine ASC were induced towards adipogenic differentiation (Monaco et al. 2009 Open Tissue Eng. Regen. Med. J. 2, 20–33) for 2 weeks in 24-well plates over round coverslips. Direct lipid profile of fatty acids and phospholipids in 3 different locations in undifferentiated and differentiated ASC was performed. Relative values of ion counts in each location were used for statistical analysis by means of a two-tailed t-test with unequal variance and false discovery rate (FDR) correction. The DIA uncovered amino acid and lipid metabolism as being the most affected pathways during adipogenesis. Among lipid-related pathways, the DIA indicated a strong induction during differentiation of unsaturation of fatty acids (FA) and metabolism of lipids, particularly triacylglycerol formation (TAG). The metabolism of tryptophan, phenylalanine, and histidine was strongly induced among amino acid-related metabolism. Among the lipid molecules identified in the mass-to-charge ratio (m/z) range of 250–1000 by the DESI-MS, >200 had significantly (FDR <0.01) lower abundance and ~30 lipids had higher abundance in differentiated compared with undifferentiated ASC. The lipid profile of undifferentiated cells was featured by a large amount of lipid molecules in the lower (m/z 780) mass range but with almost no signal in the intermediate range. The differentiated cells had a consistent large increase in amount of molecules in the range from m/z 500 to 650 and a decreased amount in the lower and upper ranges. Among the lipid molecules that decreased during differentiation, we identified several phospholipids, particularly phosphoserine and phosphoinositol species, but also linoleic, linolenic, arachidic, and eicosapentaenoic acids. The unsaturated FA oleate and docosahexaenate increased >10-fold and 2.2-fold, respectively, due to adipogenic differentiation. High-resolution experiments are being performed in order to annotate more, if not all, lipid species identified in the samples by DESI-MS. The DIA indicated greater unsaturation of FA during adipogenesis, confirmed by DESI-MS, which corresponds to the characteristic increase in lipid fractions with intermediate molecular mass. This can be considered a signature of adipogenesis in porcine ASC and essential for TAG formation. The DESI-MS appears to be a valuable tool for rapid lipid profile of cells and with the potential to be used to identify cell differentiation.

Intraoperative assessment of isocitrate dehydrogenase mutation status in human gliomas using desorption electrospray ionization–mass spectrometry

2020 ◽  
Vol 132 (1) ◽  
pp. 180-187 ◽  
Author(s):  
Clint M. Alfaro ◽  
Valentina Pirro ◽  
Michael F. Keating ◽  
Eyas M. Hattab ◽  
R. Graham Cooks ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electrospray Ionization ◽  
Isocitrate Dehydrogenase ◽  
Tumor Resection ◽  
Desorption Electrospray Ionization ◽  
Extent Of Resection ◽  
Ionization Mass ◽  
Idh Mutation ◽  
Mutation Status ◽  
Desi Ms

OBJECTIVEThe authors describe a rapid intraoperative ambient ionization mass spectrometry (MS) method for determining isocitrate dehydrogenase (IDH) mutation status from glioma tissue biopsies. This method offers new glioma management options and may impact extent of resection goals. Assessment of the IDH mutation is key for accurate glioma diagnosis, particularly for differentiating diffuse glioma from other neoplastic and reactive inflammatory conditions, a challenge for the standard intraoperative diagnostic consultation that relies solely on morphology.METHODSBanked glioma specimens (n = 37) were analyzed by desorption electrospray ionization–MS (DESI-MS) to develop a diagnostic method to detect the known altered oncometabolite in IDH-mutant gliomas, 2-hydroxyglutarate (2HG). The method was used intraoperatively to analyze tissue smears obtained from glioma patients undergoing resection and to rapidly diagnose IDH mutation status (< 5 minutes). Fifty-one tumor core biopsies from 25 patients (14 wild type [WT] and 11 mutant) were examined and data were analyzed using analysis of variance and receiver operating characteristic curve analysis.RESULTSThe optimized DESI-MS method discriminated between IDH-WT and IDH-mutant gliomas, with an average sensitivity and specificity of 100%. The average normalized DESI-MS 2HG signal was an order of magnitude higher in IDH-mutant glioma than in IDH-WT glioma. The DESI 2HG signal intensities correlated with independently measured 2HG concentrations (R2 = 0.98). In 1 case, an IDH1 R132H–mutant glioma was misdiagnosed as a demyelinating condition by frozen section histology during the intraoperative consultation, and no resection was performed pending the final pathology report. A second craniotomy and tumor resection was performed after the final pathology provided a diagnosis most consistent with an IDH-mutant glioblastoma. During the second craniotomy, high levels of 2HG in the tumor core biopsies were detected.CONCLUSIONSThis study demonstrates the capability to differentiate rapidly between IDH-mutant gliomas and IDH-WT conditions by DESI-MS during tumor resection. DESI-MS analysis of tissue smears is simple and can be easily integrated into the standard intraoperative pathology consultation. This approach may aid in solving differential diagnosis problems associated with low-grade gliomas and could influence intraoperative decisions regarding extent of resection, ultimately improving patient outcome. Research is ongoing to expand the patient cohort, systematically validate the DESI-MS method, and investigate the relationships between 2HG and tumor heterogeneity.

scholarly journals Fe(III)-doxycycline complexes with diimine ligands: Syntheses, characterization and biological properties

2019 ◽  
Vol 38 (1) ◽  
pp. 29
Author(s):  
Joshua A Obaleye ◽  
Olufunso Olumide Abosede
Keyword(s):  
Binding Constants ◽  
Biological Properties ◽  
Axial Position ◽  
Klebsiella Pneumonia ◽  
Ionization Mass ◽  
Polypyridyl Ligands ◽  
Diimine Ligands ◽  
Electrospray Ionization Mass Spectroscopy ◽  
Ionization Mass Spectroscopy ◽  
Ct Dna

Three new iron(III) complexes of doxycycline viz: [Fe(dox)2Cl]Cl2 (1), [Febpy(dox)Cl]Cl2 (2) and [Fephen(dox)Cl]Cl2 (3), where dox is doxycycline, bpy is 2,2ʹ-bipyridine and phen is 1,10-phenanthroline, were synthesized and characterized by elemental analysis, electronic absorption, FT-IR, and electrospray ionization mass spectroscopy. Doxycycline and the polypyridyl ligands behave as bidendate ligands; the polypyridyl ligands coordinate through the two diimine nitrogen atoms and doxycycline through enolate and diketoamide oxygen atoms of ring A in a five-coordinate system with chloride atom in the axial position. Their antibacterial and antiplasmodial activities against chloroquine-sensitive Plasmodium falciparum NF54 and their interaction with calf thymus (CT) DNA using electronic titration were investigated. The three complexes showed good activity against strains of Staphylococcus aureus and Klebsiella pneumonia. The complexes bind moderately to CT DNA with binding constants of 5.6 × 104 and 4.8 × 104 for complexes 2 and 3, respectively.

scholarly journals Novel Bioactive Penicipyrroether A and Pyrrospirone J from the Marine-Derived Penicillium sp. ZZ380

Marine Drugs ◽  
2019 ◽  
Vol 17 (5) ◽  
pp. 292 ◽  
Author(s):  
Tengfei Song ◽  
Mingmin Tang ◽  
Hengju Ge ◽  
Mengxuan Chen ◽  
Xiaoyuan Lian ◽  
...  
Keyword(s):  
Inhibitory Concentration ◽  
Structural Type ◽  
Ionization Mass ◽  
Isolation And Identification ◽  
Penicillium Sp ◽  
Electrospray Ionization Mass Spectroscopy ◽  
Ionization Mass Spectroscopy ◽  
Fused Ring ◽  
Ic50 Values ◽  
Fused Ring System

The marine-sourced fungus Penicillium sp. ZZ380 was previously reported to have the ability to produce a series of new pyrrospirone alkaloids. Further investigation on this strain resulted in the isolation and identification of novel penicipyrroether A and pyrrospirone J. Each of them represents the first example of its structural type, with a unique 6/5/6/5 polycyclic fusion that is different from the 6/5/6/6 fused ring system for the reported pyrrospirones. Their structures were elucidated by extensive nuclear magnetic resonance (NMR) and high resolution electrospray ionization mass spectroscopy (HRESIMS) spectroscopic analyses, electronic circular dichroism (ECD) and 13C NMR calculations and X-ray single crystal diffraction. Penicipyrroether A showed potent antiproliferative activity against human glioma U87MG and U251 cells with half maximal inhibitory concentration (IC50) values of 1.64–5.50 μM and antibacterial inhibitory activity with minimum inhibitory concentration (MIC) values of 1.7 μg/mL against methicillin-resistant Staphylococcus aureus and 3.0 μg/mL against Escherichia coli.

61 UNVEILING THE ROLE OF LIPIDS IN ORGANOGESIS: MOLECULAR ANATOMY BY DESORPTION ELECTROSPRAY IONIZATION MASS SPECTROMETRY IMAGING MASS SPECTROMETRY

2016 ◽  
Vol 28 (2) ◽  
pp. 160
Author(s):  
V. Pirro ◽  
P. O. Favaron ◽  
C. R. Ferreira ◽  
L. S. Eberlin ◽  
R. S. Barreto ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Spinal Cord ◽  
Desorption Electrospray Ionization ◽  
Ionization Mass ◽  
Differential Distribution ◽  
Desi Ms ◽  
Brain And Spinal Cord ◽  
The Brain ◽  
Digestive Glands

Even though the role of lipids in pandemic diseases such as obesity and diabetes is a focus of increasing research, the role of lipids during organogenesis, when diverse diseases may be triggered, is unexplored. Also, pig embryonic tissues represent an attractive option for organ transplantation. This study introduces a detailed morphological analysis of swine fetal tissues with matching location of lipids acquired by desorption electrospray ionization mass spectrometry (DESI-MS) imaging for the study of differential distribution of free fatty acids (FFA) and phospholipids (PL) in specific organs during fetal development. Samples from a pig fetuses around Day 50 of pregnancy were sectioned at a cryotome and mounted onto glass slides. Fixative agents were not used. DESI-MS images were run with a step size of 300 µm using a morphologically friendly (non-destructive) solvent combination, namely dimethylformamide/acetonitrile 1 : 1 (v/v). Data were acquired in the negative ion mode in the m/z range of 150 to 1000 from different sections representing the whole swine fetus body. Ion images were constructed using BioMAP software. After imaging, the whole-body tissue samples were stained with hematoxylin and eosin (H&E) and were overlaid to the DESI-MS lipid images. Differential distribution of FFA, phosphatidylcholines (PC), phosphatidylserines (PS), sulphatides (ST), and phosphatidylinositols (PI) was observed among organs, especially on nervous and circulatory systems, and digestive glands. Most lipids concentrated in the brain, spinal cord, and digestive glands such as the liver. For example, arachidonic acid was most abundant in neuronal tissue, whereas docosahexaenoic acid predominated in the liver and digestive glands. Distribution of PS (36 : 1) of m/z 788 was observed in all tissues except for the digestive system, but PS (40 : 6) of m/z 834.7 was exclusive of brain and spinal cord. Lipids related to brain and spinal cord were mostly polyunsaturated fatty acids as well as specific PS lipids. Arachidonic and eicosatrienoic acids are more concentrated in hindbrain and spinal cord, whereas PS was more abundant in the brain than in the spinal cord. There is no information on PS chemical composition during brain and spinal cord development, but PS concentration in the nervous tissue membranes varies with age, brain areas, cell type, and subcellular components. Several reports indicate that alteration in PS synthesis might participate in the mechanism of brain damage. Also, PS has been found to be altered in brain tumours. Oleic acid, fatty acid dimers, and the signalling lipid PI (38 : 3) were most significant for the digestive system and liver. Liver is one of the main organs involved in fatty acid metabolism (besides adipose tissue and muscle). By overlying morphological and molecular information, lipids seem to be a major player in the organogenesis process.

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