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- Hering, Jenny, et al.
(författare)
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The rapid "teabag" method for high-end purification of membrane proteins
- 2020
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 10:1
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Tidskriftsartikel (refereegranskat)abstract
- Overproduction and purification of membrane proteins are generally challenging and time-consuming procedures due to low expression levels, misfolding, and low stability once extracted from the membrane. Reducing processing steps and shortening the timespan for purification represent attractive approaches to overcome some of these challenges. We have therefore compared a fast "teabag" purification method with conventional purification for five different membrane proteins (MraY, AQP10, CIC-1, PAR2 and KCC2). Notably, this new approach reduces the purification time significantly, and the quality of the purified membrane proteins is equal to or exceeds conventional methods as assessed by size exclusion chromatography, SDS-PAGE and downstream applications such as ITC, crystallization and cryo-EM. Furthermore, the method is scalable, applicable to a range of affinity resins and allows for parallelization. Consequently, the technique has the potential to substantially simplify purification efforts of membrane proteins in basic and applied sciences.
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| 2. |
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| 3. |
- Kitchen, Philip, et al.
(författare)
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Targeting Aquaporin-4 Subcellular Localization to Treat Central Nervous System Edema
- 2020
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Ingår i: Cell. - : Elsevier BV. - 0092-8674. ; 181:4, s. 19-799
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Tidskriftsartikel (refereegranskat)abstract
- Swelling of the brain or spinal cord (CNS edema) affects millions of people every year. All potential pharmacological interventions have failed in clinical trials, meaning that symptom management is the only treatment option. The water channel protein aquaporin-4 (AQP4) is expressed in astrocytes and mediates water flux across the blood-brain and blood-spinal cord barriers. Here we show that AQP4 cell-surface abundance increases in response to hypoxia-induced cell swelling in a calmodulin-dependent manner. Calmodulin directly binds the AQP4 carboxyl terminus, causing a specific conformational change and driving AQP4 cell-surface localization. Inhibition of calmodulin in a rat spinal cord injury model with the licensed drug trifluoperazine inhibited AQP4 localization to the blood-spinal cord barrier, ablated CNS edema, and led to accelerated functional recovery compared with untreated animals. We propose that targeting the mechanism of calmodulin-mediated cell-surface localization of AQP4 is a viable strategy for development of CNS edema therapies.
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| 4. |
- Lousto, Carlos O., et al.
(författare)
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PULSAR OBSERVATIONS AT LOW LATITUDES AND LOW FREQUENCIES
- 2024
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Ingår i: Revista Mexicana de Astronomia y Astrofisica: Serie de Conferencias. - 1405-2059. ; 56, s. 134-144
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Konferensbidrag (refereegranskat)abstract
- The Pulsar Monitoring in Argentina (PuMA) is a collaboration between the Argentine Institute for Radioastronomy (IAR) and the Rochester Institute of Technology (RIT) that since 2017 has been observing southern sky pulsars with high cadence using the two restored IAR antennas in the L-Band (1400 MHz). We briefly review the first set of results of this program to study transient phenomena, such as magnetars and glitching pulsars, as well as to perform precise timing of millisecond pulsars. Access to lower frequency bands, where most of the pulsars are brighter, would allow us to reach additional pulsars, currently buried into the background noise. We identify two dozen additional glitching pulsars that could be observable in the 400 MHz band by the IAR’s projected Multipurpose Interferometer Array (MIA). We also discuss the relevance and challenges of single-pulse pulsar timing at low frequencies and the search for Fast Radio Burst (FRB) in the collected data since 2017 using machine learning techniques.
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| 5. |
- Zubieta, Ezequiel, et al.
(författare)
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First results of the glitching pulsar monitoring programme at the Argentine Institute of Radioastronomy
- 2023
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Ingår i: Monthly Notices of the Royal Astronomical Society. - : Oxford University Press (OUP). - 0035-8711 .- 1365-2966. ; 521:3, s. 4504-4521
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Tidskriftsartikel (refereegranskat)abstract
- We report here on the first results of a systematic monitoring of southern glitching pulsars at the Argentine Institute of Radioastronomy that started in the year 2019. We detected a major glitch in the Vela pulsar (PSR J0835 - 4510) and two small glitches in PSR J1048 - 5832. For each glitch, we present the measurement of glitch parameters by fitting timing residuals. We then make an individual pulse study of Vela in observations before and after the glitch. We selected 6 days of observations around the major glitch on 2021 July 22 and study their statistical properties with machine learning techniques. We use variational autoencoder (VAE) reconstruction of the pulses to separate them clearly from the noise. We perform a study with self-organizing map (SOM) clustering techniques to search for unusual behaviour of the clusters during the days around the glitch not finding notable qualitative changes. We have also detected and confirmed recent glitches in PSR J0742 - 2822 and PSR J1740 - 3015.
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