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Hypoxia-induced long non-coding RNA Malat1 is dispensable for renal ischemia/reperfusion-injury
(2018)
Renal ischemia-reperfusion (I/R) injury is a major cause of acute kidney injury (AKI). Non-coding RNAs are crucially involved in its pathophysiology. We identified hypoxia-induced long non-coding RNA Malat1 (Metastasis Associated Lung Adenocarcinoma Transcript 1) to be upregulated in renal I/R injury. We here elucidated the functional role of Malat1 in vitro and its potential contribution to kidney injury in vivo. Malat1 was upregulated in kidney biopsies and plasma of patients with AKI, in murine hypoxic kidney tissue as well as in cultured and ex vivo sorted hypoxic endothelial cells and tubular epithelial cells. Malat1 was transcriptionally activated by hypoxia-inducible factor 1-α. In vitro, Malat1 inhibition reduced proliferation and the number of endothelial cells in the S-phase of the cell cycle. In vivo, Malat1 knockout and wildtype mice showed similar degrees of outer medullary tubular epithelial injury, proliferation, capillary rarefaction, inflammation and fibrosis, survival and kidney function. Small-RNA sequencing and whole genome expression analysis revealed only minor changes between ischemic Malat1 knockout and wildtype mice. Contrary to previous studies, which suggested a prominent role of Malat1 in the induction of disease, we did not confirm an in vivo role of Malat1 concerning renal I/R-injury.
Ammonium CI-Orbitrap: a tool for characterizing the reactivity of oxygenated organic molecules
(2024)
Oxygenated organic molecules (OOMs) play an important role in the formation of atmospheric aerosols. Due to various analytical challenges with respect to measuring organic vapors, uncertainties remain regarding the formation and fate of OOMs. The chemical ionization Orbitrap (CI-Orbitrap) mass spectrometer has recently been shown to be a powerful technique that is able to accurately identify gaseous organic compounds due to its greater mass resolution. Here, we present the ammonium-ion-based CI-Orbitrap (-Orbitrap) as a technique capable of measuring a wide range of gaseous OOMs. The performance of the -Orbitrap is compared with that of state-of-the-art mass spectrometers, including a nitrate-ion-based chemical ionization atmospheric pressure interface coupled to a time-of-flight mass spectrometer (-LTOF), a new generation of proton transfer reaction-TOF mass spectrometer (PTR3-TOF), and an iodide-based CI-TOF mass spectrometer equipped with a Filter Inlet for Gases and AEROsols (I−-CIMS). The instruments were deployed simultaneously in the Cosmic Leaving OUtdoors Droplets (CLOUD) chamber at the European Organization for Nuclear Research (CERN) during the CLOUD14 campaign in 2019. Products generated from α-pinene ozonolysis under various experimental conditions were simultaneously measured by the mass spectrometers. The -Orbitrap was able to identify the widest range of OOMs (i.e., O ≥ 2), from less-oxidized species to highly oxygenated organic molecules (HOMs). Excellent agreement was found between the -Orbitrap and the -LTOF with respect to characterizing HOMs and with the PTR3-TOF for the less-oxidized monomeric species. OOM concentrations measured by -Orbitrap were estimated using calibration factors derived from the OOMs with high time-series correlations during the side-by-side measurements. As with the other mass spectrometry techniques used during this campaign, the detection sensitivity of the -Orbitrap to OOMs is greatly affected by relative humidity, which may be related to changes in ionization efficiency and/or multiphase chemistry. Overall, this study shows that -ion-based chemistry associated with the high mass resolution of the Orbitrap mass analyzer can measure almost all inclusive compounds. As a result, it is now possible to cover the entire range of compounds, which can lead to a better understanding of the oxidation processes.
Ammonium CI-Orbitrap: a tool for characterizing the reactivity of oxygenated organic molecules
(2023)
Oxygenated organic molecules (OOMs) play an important role in the formation of atmospheric aerosols. Due to various analytical challenges in measuring organic vapors, uncertainties remain in the formation and fate of OOMs. The chemical ionization Orbitrap mass spectrometer (CI-Orbitrap) has recently been shown to be a powerful technique able to accurately identify gaseous organic compounds due to its great mass resolving power. Here we present the ammonium ion (NH4+) based CI-Orbitrap as a technique capable of measuring a wide range of gaseous OOMs. The performance of the CI-(NH4+)-Orbitrap was compared with that of state-of-the-art mass spectrometers, including a nitrate ion (NO3−) based CI coupled to an atmospheric pressure interfaced to long time-of-flight mass spectrometer (APi-LTOF), a new generation of proton transfer reaction-TOF mass spectrometer (PTR3-TOF), and an iodide (I−) based CI-TOF mass spectrometer equipped with a Filter Inlet for Gases and AEROsols (FIGAERO-CIMS). The instruments were deployed simultaneously in the Cosmic Leaving OUtdoors Droplets (CLOUD) chamber at the European Organization for Nuclear Research (CERN) during the CLOUD14 campaign in 2019. Products generated from α-pinene ozonolysis across multiple experimental conditions were simultaneously measured by the mass spectrometers. NH4+-Orbitrap was able to identify the widest range of OOMs (i.e., O ≥ 2), from low oxidized species to highly oxygenated volatile organic compounds (HOM). Excellent agreements were found between the NH4+-Orbitrap and the NO3−-LTOF for characterizing HOMs and with the PTR3-TOF for the less oxidized monomeric species. A semi-quantitative information was retrieved for OOMs measured by NH4+-Orbitrap using calibration factors derived from this side-by-side comparison. As other mass spectrometry techniques used during this campaign, the detection sensitivity of NH4+-Orbitrap to OOMs is greatly affected by relative humidity, which may be related to changes in ionization efficiency and/or multiphase chemistry. Overall, this study shows that NH4+ ion-based chemistry associated with the high mass resolving power of the Orbitrap mass analyzer can measure almost all-inclusive compounds. As a result, it is now possible to cover the entire range of compounds, which can lead to a better understanding of the oxidation processes.
