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Abstract: Hypoxia refers to a condition where cells and tissues experience low, inadequate levels of O2. While healthy tissues are typically supplied with sufficient O2 (normoxia), cancerous tissues commonly face hypoxia due to the tumor’s extraordinarily high demand for oxygen. Various fluorescent small-molecule probes have been designed for selective detection of hypoxia in living cells, but few nanomaterials have been investigated for this type of bioimaging. Herein, we prepare a fluorescent covalent organic framework (COF) with β-ketoenamine linkages and post-synthetically modify it to conjugate hypoxia-sensitive nitroimidazole moieties into its pores (NI-COF). Stacks of sheets in NI-COF observed under electron microscopy were exfoliated by ultrasonication, and dynamic light scattering measurements confirmed particle size of less than 200 nm. Thus-prepared material exhibited good stability in physiological conditions and low cytotoxicity in in vitro experiments. NI-COF also showed useful fluorescence properties with an emission peak at 490 nm (λex = 420 nm) at both neutral and mildly acidic pH levels that are characteristic of tumor tissues. Encouraged by the favorable properties of the material, we incubated HeLa cells pre-treated in either hypoxic or normoxic conditions with NI-COF. Fluorescence microscopy images demonstrated that the material was preferentially taken up by hypoxic cells, which showed higher fluorescence signal in their interior than cells cultured under normoxia conditions. It is anticipated that this study will stimulate further developments of COFs for imaging various biological conditions.
Keywords: hypoxia, fluorescence, covalent organic frameworks, imaging, tumor cells
Published in RUNG: 19.09.2023; Views: 116; Downloads: 1
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Keywords: covalent organic frameworks, imaging, hypoxia, fluorescence, cancer cells
Published in RUNG: 17.05.2023; Views: 410; Downloads: 6
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Keywords: HPV, APOBEC proteins, gene expression, TCGA data
Published in RUNG: 29.11.2022; Views: 614; Downloads: 31
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Published in RUNG: 14.09.2022; Views: 684; Downloads: 7
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Abstract: Different analytical methods were tested and optimized for the determination of four biogenic amines (BA) histamine, putrescine, cadaverine and tyramine produced by grape-associated lactic acid bacteria (LAB). Autochthonous LAB were isolated from ‘Refošk’ grapes belonging to Slovenian-Italian Karst region as they represent a potential pool of beneficial LAB starter bacteria for improving the typicality and quality of wine. Six isolated strains were screened by multiplex PCR, of which four strains were positive for BA-forming genes (MKBT-49, MKBT-282, MKBT-568, MKBT-570). The production of BA was evaluated by thin-layer chromatography (TLC), high-performance liquid chromatography coupled to diode array detection (HPLC-DAD) and enzymatic method. The HPLC-DAD results showed that strain MKBT-49 (tyrdc+) had high tyramine production (386.6 ± 0.14 mg/L), which was confirmed by TLC. The ability to produce putrescine was confirmed in strain MKBT-282 by PCR, HPLC-DAD (16.4 ± 1.72 mg/L), and TLC. Histamine-producing ability was detected in strain MKBT-570, with a concentration below the limit of detection of the HPLC-DAD (<0.2 mg/L), while the other two methods were not sensitive enough for confirmation. This study shows that the production of BA can detected in native LAB and that relatively simple method such as TLC can be used effectively for the initial screening.
Keywords: grapes, wine-like matrices, biogenic amines, autochthonous lactic acid bacteria, analytical methods
Published in RUNG: 03.01.2022; Views: 1151; Downloads: 6
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