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- Cossarizza, A., et al.
(author)
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Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
- 2019
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In: European Journal of Immunology. - : Wiley. - 0014-2980 .- 1521-4141. ; 49:10, s. 1457-1973
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Journal article (peer-reviewed)abstract
- These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.
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- Semba, H, et al.
(author)
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HIF-1α-PDK1 axis-induced active glycolysis plays an essential role in macrophage migratory capacity
- 2016
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In: Nature communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 7, s. 11635-
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Journal article (peer-reviewed)abstract
- In severely hypoxic condition, HIF-1α-mediated induction of Pdk1 was found to regulate glucose oxidation by preventing the entry of pyruvate into the tricarboxylic cycle. Monocyte-derived macrophages, however, encounter a gradual decrease in oxygen availability during its migration process in inflammatory areas. Here we show that HIF-1α-PDK1-mediated metabolic changes occur in mild hypoxia, where mitochondrial cytochrome c oxidase activity is unimpaired, suggesting a mode of glycolytic reprogramming. In primary macrophages, PKM2, a glycolytic enzyme responsible for glycolytic ATP synthesis localizes in filopodia and lammelipodia, where ATP is rapidly consumed during actin remodelling processes. Remarkably, inhibition of glycolytic reprogramming with dichloroacetate significantly impairs macrophage migration in vitro and in vivo. Furthermore, inhibition of the macrophage HIF-1α-PDK1 axis suppresses systemic inflammation, suggesting a potential therapeutic approach for regulating inflammatory processes. Our findings thus demonstrate that adaptive responses in glucose metabolism contribute to macrophage migratory activity.
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| 7. |
- Badea, M, et al.
(author)
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A flow immunoassay for alkylphenol ethoxylate surfactants and their metabolites - questions associated with cross-reactivity, matrix effects, and validation by chromatographic techniques
- 2003
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In: Analyst. - : Royal Society of Chemistry (RSC). - 1364-5528. ; 128:7, s. 849-856
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Journal article (peer-reviewed)abstract
- This paper describes the application and evaluation of a competitive enzyme flow injection immunoassay (EFIIA) for screening of alkylphenol ethoxylate (APEO) surfactants in different water samples based on a generic immunoassay system previously developed ( see E. Burestedt, C Nistor, U. Schagerlof and J. Emneus, Anal. Chem., 2000, 72, 4171 - 4177). The detection limits for octylphenol ethoxylates (OPEOs), nonylphenol ethoxylates (NPEOs), and nonylphenol (NP) were 0.5 mug l(-1), between 2 and 3 mug l(-1), and 50 mug l(-1), respectively, with a sample throughput of 6 h(-1) (i.e., for triplicate analysis of each sample). Different OPEOs and NPEOs were highly cross-reactive within the assay, with sensitivities in the same order of magnitude for all the ethoxylates tested, thus the result obtained by the EFIIA method could be used as an "alkylphenol ethoxylate index". No or minor matrix effects with recoveries between 70 - 120% for the reference analyte NPEO10 in tap, and surface water, and acceptable for rainwater, were observed. Influent and effluent surfactant containing wastewater samples were analysed by EFIIA, LC-MS, LC-Fluoresence (LC-FL), and a commercial microplate ELISA. High recoveries for different concentrations of APEO(10) spiked into a 200 times diluted raw influent and effluent wastewater were achieved with the EFIIA method, however, the found APEO content of the same diluted wastewater samples, before spiking, could not be correlated directly to the chromatographic result by any of the immunoassays, and the possible reasons for this are discussed. The same trend of decreasing APEO content from influent to effluent wastewater could, however, be followed for all methods employed.
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