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Keywords = enzyme-assisted derivatization

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15 pages, 3754 KB  
Article
Anti-Osteoporotic Effect of Viscozyme-Assisted Polysaccharide Extracts from Portulaca oleracea L. on H2O2-Treated MC3T3-E1 Cells and Zebrafish
by Yunhua Fu, Xuan Hu, Dongyue Zhou, Xue Li, Xingyu Tao, Di Yang, Fei Zheng, Yulin Dai and Hao Yue
Separations 2022, 9(5), 128; https://doi.org/10.3390/separations9050128 - 20 May 2022
Cited by 6 | Viewed by 2845
Abstract
This study aims to screen and characterize the protective effect of polysaccharides from Portulaca oleracea L. (POP) against H2O2-stimulated osteoblast apoptosis in vivo and in vitro. The enzymes viscozyme, celluclast, α-amylase, and β-glucanase were used to extract POPs. Among [...] Read more.
This study aims to screen and characterize the protective effect of polysaccharides from Portulaca oleracea L. (POP) against H2O2-stimulated osteoblast apoptosis in vivo and in vitro. The enzymes viscozyme, celluclast, α-amylase, and β-glucanase were used to extract POPs. Among all enzyme-assisted POPs, the first participating fraction of viscozyme extract POP (VPOP1) exhibited the highest antioxidant activity. Hoechst 33342 and acridine orange/ethidium bromide staining and flow cytometry of MC3T3 cells revealed that VPOP1 inhibited apoptosis in a dose-dependent manner. Moreover, VPOP1 increased the expression levels of heme oxygenase-1 (HO-1) and NADPH quinine oxidoreductase 1 (NQO1) and decreased the expression levels of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and Kelch-like ECH-associated protein 1 (Keap1) in H2O2-induced cells compared with their controls. The results of an in vivo experiment show that VPOP1 significantly reduced reactive oxygen species generation and lipid peroxidation in zebrafish at 72 h post-fertilization and promoted bone growth at 9 days post-fertilization. Furthermore, VPOP1 was identified via 1-phenyl-3-methyl-5-pyrazolone derivatization as an acidic heteropolysaccharide comprising mannose and possessing a molecular weight of approximately 7.6 kDa. Collectively, VPOP1 was selected as a potential anti-osteoporotic functional food because of its protective activity against H2O2-induced damage in vitro and in vivo. Full article
(This article belongs to the Special Issue Hyphenated Techniques’ Enlightenment in Chemical Analysis: From Complex Matrices of Plant Extracts and Foods to Human and Human Metabolism)
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21 pages, 4454 KB  
Article
A Multi-Analytical Approach for the Characterization of Seventeenth Century Decorative Wall Paintings in Two Norwegian Stave Churches: A Case Study at Eidsborg and Heddal, Norway
by Ashley Amanda Freeman, Lavinia de Ferri, Joy Mazurek, Fabrizio Andriulo and Chiara Bertolin
Appl. Sci. 2021, 11(8), 3477; https://doi.org/10.3390/app11083477 - 13 Apr 2021
Cited by 6 | Viewed by 4379
Abstract
The presented research examines 17th century distemper paint from the polychrome wooden interiors of two Norwegian stave churches: Eidsborg and Heddal. For the first time, the inorganic and organic components of specimens from Eidsborg and Heddal were identified using X-ray Diffraction (XRD), Environmental [...] Read more.
The presented research examines 17th century distemper paint from the polychrome wooden interiors of two Norwegian stave churches: Eidsborg and Heddal. For the first time, the inorganic and organic components of specimens from Eidsborg and Heddal were identified using X-ray Diffraction (XRD), Environmental Scanning Electron Microscopy (ESEM)—Energy Dispersive X-ray Spectroscopy (EDS), Fourier-Transform Infrared (FT-IR) spectroscopy, Enzyme-Linked Immunosorbent Assay (ELISA), and Gas Chromatography-Mass Spectrometry (GC-MS) after derivatization. This multi-analytical approach allowed for the identification of red ochre as the main red pigment within the topcoat (with the possible addition of minium), confirmed that a chalk basecoat was used, and finally permitted the recognition of alteration phases. Markers of proteinaceous material attributed to the use of animal-based glues were detected throughout the stratigraphic layers of both churches, with the addition of linseed oil in some locations. Furthermore, the wood substrate showed markers characteristic of pine tree, with contamination of wood fractions being detected in some of the paint samples from Heddal and Eidsborg. This research has contributed to a better understanding of the current preservation state of Heddal and Eidsborg, and ultimately assisted in developing a deeper comprehension and awareness of materials used in Norwegian stave churches. Full article
(This article belongs to the Special Issue Scientific Methods for Cultural Heritage)
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20 pages, 5636 KB  
Communication
Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry
by Jonas Abdel-Khalik, Peter J. Crick, Eylan Yutuc, Yuqin Wang and William J. Griffiths
Molecules 2019, 24(3), 597; https://doi.org/10.3390/molecules24030597 - 7 Feb 2019
Cited by 2 | Viewed by 5658
Abstract
Enzyme-assisted derivatization for sterol analysis (EADSA) is a technology designed to enhance sensitivity and specificity for sterol analysis using electrospray ionization–mass spectrometry. To date it has only been exploited on sterols with a 3β-hydroxy-5-ene or 3β-hydroxy-5α-hydrogen structure, using bacterial cholesterol oxidase enzyme to [...] Read more.
Enzyme-assisted derivatization for sterol analysis (EADSA) is a technology designed to enhance sensitivity and specificity for sterol analysis using electrospray ionization–mass spectrometry. To date it has only been exploited on sterols with a 3β-hydroxy-5-ene or 3β-hydroxy-5α-hydrogen structure, using bacterial cholesterol oxidase enzyme to convert the 3β-hydroxy group to a 3-oxo group for subsequent derivatization with the positively charged Girard hydrazine reagents, or on substrates with a native oxo group. Here we describe an extension of the technology by substituting 3α-hydroxysteroid dehydrogenase (3α-HSD) for cholesterol oxidase, making the method applicable to sterols with a 3α-hydroxy-5β-hydrogen structure. The 3α-HSD enzyme works efficiently on bile alcohols and bile acids with this stereochemistry. However, as found by others, derivatization of the resultant 3-oxo group with a hydrazine reagent does not go to completion in the absence of a conjugating double bond in the sterol structure. Nevertheless, Girard P derivatives of bile alcohols and C27 acids give an intense molecular ion ([M]+) upon electrospray ionization and informative fragmentation spectra. The method shows promise for analysis of bile alcohols and 3α-hydroxy-5β-C27-acids, enhancing the range of sterols that can be analyzed at high sensitivity in sterolomic studies. Full article
(This article belongs to the Special Issue Chemical Biology of Sterols, Triterpenoids and Other Natural Products: A Themed Issue in Honor of Professor W. David Nes on the Occasion of His 65th Birthday)
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24 pages, 983 KB  
Article
High Levels of Structural Diversity Observed in Microcystins from Microcystis CAWBG11 and Characterization of Six New Microcystin Congeners
by Jonathan Puddick, Michèle R. Prinsep, Susanna A. Wood, Sangata A. F. Kaufononga, Stephen Craig Cary and David P. Hamilton
Mar. Drugs 2014, 12(11), 5372-5395; https://doi.org/10.3390/md12115372 - 13 Nov 2014
Cited by 171 | Viewed by 11176
Abstract
Microcystins (MCs) are cyclic peptides produced by cyanobacteria, which can be harmful to humans and animals when ingested. Differences in the coding of the non‑ribosomal peptide synthetase/polyketide synthase enzyme complex responsible for microcystin production have resulted in more than 100 microcystin variants being [...] Read more.
Microcystins (MCs) are cyclic peptides produced by cyanobacteria, which can be harmful to humans and animals when ingested. Differences in the coding of the non‑ribosomal peptide synthetase/polyketide synthase enzyme complex responsible for microcystin production have resulted in more than 100 microcystin variants being reported to date. The microcystin diversity of Microcystis CAWBG11 was investigated using matrix-assisted laser desorption/ionization-time of flight mass spectrometry and liquid chromatography-mass spectrometry. This revealed that CAWBG11 simultaneously produced 21 known microcystins and six new congeners: [Asp3] MC-RA, [Asp3] MC-RAba, [Asp3] MC-FA, [Asp3] MC-WA, MC-FAba and MC-FL. The new congeners were putatively characterized by tandem mass spectrometry and chemical derivatization. A survey of the microcystin congeners produced by 49 cyanobacterial strains documented in scientific literature showed that cyanobacteria generally produce four microcystin congeners, but strains which produce up to 47 microcystin congeners have been reported. Microcystis CAWBG11 (which produces at least 27 congeners) was positioned in the top ten percentile of the strains surveyed, and showed fluidity of the amino acids incorporated into both position two and position four. Full article
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