Resultados de investigación

This study analyzed the variability of equivalent black carbon (eBC) mass concentrations and their sources in urban Europe to provide insights into the use of eBC as an advanced air quality (AQ) parameter for AQ standards. This study compiled eBC mass concentration datasets covering the period between 2006 to 2022 from 50 measurement stations, including 23 urban background (UB), 18 traffic (TR), 7 suburban (SUB), and 2 regional background (RB) sites. The results highlighted the need for the harmonization of eBC measurements to allow for direct comparisons between eBC mass concentrations measured across urban Europe. The eBC mass concentrations exhibited a decreasing trend as follows: TR > UB > SUB > RB. Furthermore, a clear decreasing trend in eBC concentrations was observed in the UB sites moving from Southern to Northern Europe. The eBC mass concentrations exhibited significant spatiotemporal heterogeneity, including marked differences in eBC mass concentration and variable contributions of pollution sources to bulk eBC between different cities. Seasonal patterns in eBC concentrations were also evident, with higher winter concentrations observed in a large proportion of cities, especially at UB and SUB sites. The contribution of eBC from liquid fossil fuel combustion, mostly traffic (eBCT) was higher than that of residential and commercial sources (eBCRC) in all European sites studied. Nevertheless, eBCRC still had a substantial contribution to total eBC mass concentrations at a majority of the sites. eBC trend analysis revealed decreasing trends for eBCT over the last decade, while eBCRC remained relatively constant or even increased slightly in some cities., Attention: Use of data may require contact with data provider. Please request acknowledgement details from data originators in compliance with data sharing and citation. Attribution to RI-URBANS is required from users to give appropriate credit. Information on this is included in the files. All scientific articles and reports using data need to acknowledge the project; "This study is using data that has received funding from the European Union's Horizon 2020 research and innovation programm under grant agreement No 101036245". Grants: Agencia Estatal de Investigación, Spanish Ministry of Science and Innovation, CAIAC project (PID2019-108990RB-I00); Spanish Ministry of Science and Innovation, FPI grant (PRE-2020-095498); Generalitat de Catalunya (AGAUR, SGR-447), Peer reviewed

NMR datasets used in https://doi.org/10.1021/acs.analchem.8b01196. In the corresponding study, we have performed a comparative chemometric analysis between untargeted 1H NMR and 1H-13C HSQC NMR analyses of metabolomics samples from Saccharomyces cerevisiae (yeast) extracts. Specifically, yeast was grown in two different liquid media and their metabolism was characterized at 8 different time-points of a 3-day period. The two media used, YPD (Yeast Peptone Dextrose) and YSC (Yeast nitrogen base Synthetic Complete), are broadly used in yeast lab routines, and results from this analysis should be of interest for improving lab methodologies involving yeast., Peer reviewed

The exposure protocol involved zebrafish embryos exposed in groups of 20 to various concentrations of chemical compounds, with five replicates per treatment. The concentrations ranged from the lowest observed effect concentrations (LOECs) to control levels. After exposure, embryos were collected, washed, frozen, and stored. Metabolites were extracted from individual embryo pools using methanol and methionine sulfone. The extraction process included vortexing, sonication, and centrifugation, followed by addition of water and chloroform. The aqueous fraction was dried and reconstituted using acetonitrile-water solution. Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) was used for analysis. Chromatographic separations were carried out on a hydrophilic interaction liquid chromatography (HILIC) column. Mass spectrometry was performed using an Orbitrap mass spectrometer with electrospray ionization in positive and negative modes. The mass spectra were acquired at high resolution, and fragmentation scans were used for metabolite identification. The overall process aimed to analyze the metabolomic profile of zebrafish embryos exposed to different chemical concentrations., Peer reviewed

The dataset includes two Excel spreadsheets with the soil and water characteristics from samples obtained during wet (April) and dry (September) seasons in 2019, and at the end of the experiment, July 2020. The sampling was designed to characterize the soil and water in the context of the study of soil redox conditions using IRIS (Indicators of Reducing of Soils) devices. The selected sampling locations represent a gradient of water within the playa-lake: bare soil very frequently flooded (SAL-1); bare soil often flooded (SAL-2); and the plot colonized by annual and perennial halophytes (SAL-3). Soil data includes the main description of the soil layers per date in the three sampling plots, and their chemical and physical analytical data determined in the field and in the lab. We analyzed water from three different origins: i) surface water when present, ii) shallow groundwater from piezometers up to 80 cm depth, iii) suction probes installed at 20 and 40 cm depth, and iv) saturated paste extracts of soil samples. Photograph P1170434.jpg shows a view of the field setting of the experiment in 2019, April 17., The data corresponds to the Spanish study area, the Salineta inland saline wetland (41°28'55.34"N, 0° 9'23.59"W) located 60 km far from Zaragoza city, in one of the most arid areas of the Central Ebro Basin, NE Spain. In this project we studied the redox conditions of the hypersaline soils in Salineta playa-lake, which are subjected to intermittent flooding, along a period of 17 months, in 2019 and 2020., These data belong to the research project AQUASALT, EraNET that in Spain has been supported by grant PCI2018-09299 funded by MCIN/AEI/10.13039/50110 0 011033 and co-funded by the European Union., Peer reviewed

(A) CBK1 cbk1Δ cdc15-2 (YMF3869), (B) cbk1-6E cbk1Δ cdc15-2 (YMF3763), (C) cbk1-5E-E409S cbk1Δ cdc15-2 (YMF4279), (D) cbk1-5E-E574T cbk1Δ cdc15-2 (YMF4280), and (E) cbk1-9A cbk1Δ cdc15-2 (YMF3764) cells were grown in YPD and arrested in late anaphase by raising the temperature to 37 °C before rapamycin was added to half of the culture for 20 min. Subsequently, to allow progression through the cell cycle, cells were released in the absence (i) or presence (ii) of rapamycin. Samples were taken at the indicated times to study cell-cycle progression by flow cytometry. Schematic illustration of cbk1-9A mutant in which phosphosites containing serines or threonines followed by prolines were changed to alanine to block phosphorylations (iii). FACS graphs can be found in the supplementary FACS file (S1 File)., pbio.3002263.s006.pdf, Peer reviewed

(A) DSE4-6HA cdc15-2 (YMF4029) cells were grown in YPD and arrested in late anaphase by shifting the temperature to 37 °C before the addition of DMSO (−) or rapamycin (+) for 20 min. Subsequently, cells were released from the anaphase arrest in the absence (−) or presence (+) of rapamycin before protein extracts were prepared from shown time points and analysed by immunoblotting. Raw data for blot can be found in Supporting information (S6A Raw Images). (B) iHA-CTS1 cdc14-1 cells (YMF4088) were grown and processed as in A. Protein extracts were prepared from indicated time points and analysed by immunoblotting. Raw data for blot can be found in Supporting information (S6B Raw Images). (C) CTS1-GFPEnvy cdc14-1 cells (YMF4231) were grown as in C. Samples were taken at the indicated times to determine the proportion of cells with Cts1 at the division site in the presence (i) of rapamycin. Examples of cells are shown for the 60 min time point after the release at 24 °C in the presence (ii) of rapamycin. Scale bar indicates 5 μm. Underlying data for all the graphs can be found in S1 Data file., pbio.3002263.s007.pdf, Peer reviewed

1 -xlsx File (19.0 kB). Under Commons Attribution 4.0 International., Field data from four Mediterranean shrublands monitored during an extreme drought in 2014 (hydrological year) in SE Spain. Data are plant dieback of the total plant community and main plant biotypes (shrubs, subshrubs and grasses), vegetation structure attributes (phytovolume, plant height, LAI, and sizes of patch and inter-patch areas), soil properties (soil depth and soil surface cover types) and water availability indicators (soil moisture and environmental variables)., Peer reviewed

(A) CBK1-GFP cdc15-2 (YMF3566) and CBK1-GFP TOR1-1 cdc15-2 (YMF3565) cells were grown in YPD and arrested in late anaphase by shifting the temperature to 37 °C before the addition of rapamycin for 20 min to both strains. In addition, DMSO was added to one half of CBK1-GFP cdc15-2. Untagged cdc15-2 (YMF3580) and TOR1-1 cdc15-2 cells (YMF3578) were used as controls. Cell extracts were prepared before the immunoprecipitation of Cbk1-GFP on ChromoTek GFP-Trap Magnetic beads. Isolated material was subjected to analysis by mass spectrometry. Numbers represent spectral average of key Cbk1 functional-related interactors in 3 independent replicates of the experiment. The average is calculated as the sum of all spectral counts for the 3 replicates divided by 3. We found Cbk1 interactions previously described: Mob2 [48] or components of the RAM pathway such as Tao3 [97], Sog2 [98], and Kic1 [99]. Besides, Cbk1 negative regulators Lre1 and Fir1 were also identified [5,43]. Besides, MS found Myo1, a component of the actomyosin ring [43]. Our results showed that Cbk1 interacts with the transcription factor Ace2 [47,48]. Moreover, Cbk1 was bound to Ssd1, an RNA-binding protein that represses the translation of cell wall remodelling proteins until Ssd1 is phosphorylated by Cbk1 [48,100]. Interestingly, TORC1 and Ssd1 have been described to collaborate to maintain cellular integrity [73]. We found interactions with paralogs serine/threonine kinases Kin1 and Kin2 [74,75]. Finally, we detected protein Yol036w of unknown function interacting with Cbk1 [40,74]. Details of other known Cbk1 interactors are shown in S2 Table. (B) CBK1-5FLAG ACE2-HA cdc15-2 (YMF3585) and control cells (YMF3587) (i), CBK1-5FLAG MOB2-9MYC cdc15-2 (YMF3838) and control cells (YMF3835) (ii), or CBK1-TAP SOG2-6HA cdc15-2 (YMF4235) and control cells (YMF4236) cells (iii) were grown in YPD and arrested in late anaphase by shifting to 37 °C before the addition of rapamycin for 20 min when indicated. Subsequently, protein extracts were prepared and immunoprecipitations of Cbk1 on FLAG-beads or IgG beads (as denoted) were performed before the detection of the indicated proteins by immunoblotting. Raw data for blots can be found in Supporting information (S7 Raw Images). (C) Components of TORC1 were found in the same mass spectrometry analysis as in A. Numbers represent spectral average of 3 independent replicates of the experiment., pbio.3002263.s008.pdf, Peer reviewed

(A) Orthologues of S. cerevisiae Sec3 in the indicated fungal species were identified by PSI-BLAST searches, and a CLUSTAL O multiple sequence alignment was performed. Conserved NDR/LATS phosphorylation consensus is marked with a red square following consensus in S. cerevisiae [47] and in S. pombe [54]. Putative Cbk1 phosphorylation sites were identified in Sec3 by scanning the sequence (S18, S32, S43, and S66). (B) To confirm that Sec3 was essential in the budding yeast strain used in this work, we deleted SEC3 in diploid cells (YMF4159). Spores that lack Sec3 were able to grow extremely deficiently, although they formed tiny colonies (i). To conditionally inactivate Sec3, we planned to follow the same strategy as we did for the study of Cbk1 mutants that is explained in detail in S2E Fig. We added the “auxin inducible degron” (“aid”) cassette [38] to the C-terminal end of SEC3. Then, control cells (ADH-TIR1; YJW15) and sec3-aid ADH-TIR1 cells (YMF4612) were grown at 24 °C on YPD medium before serial dilutions of 50,000, 5,000, 500, and 50 cells were plated on the indicated media and incubated for 3 days (ii). sec3-aid ADH-TIR1 cells were able to grow under restrictive conditions at 24 °C (ii), which showed that Sec3 depletion was not completely effective. This prevented us from the use of sec3-aid mutant, in combination with cdc15-2 to inactivate Cdc15 function at 37 °C, and test whether phosphomimetic version of Sec3 (Sec3-4E, S18E, S32E, S43E, and S66E) was able to rescue cell separation defects associated to cdc15-2 cells. (C) To be able to follow version of t-SNARE Sso1 that lacks N-terminal autoinhibition domain, we deleted DNA sequence that corresponds to amino acids 1–146. At the same time, we fused the tag 5xFLAG at the N-terminal of the truncated Sso1 to be able to follow the protein dynamics. As control, we tagged the N-terminal end of wt Sso1 with 5xFLAG. To determine whether 5xFLAG-Sso1 and 5xFLAG-Sso1Δ1–146 were functional, we combined 5xFLAG-SSO1 (i) or 5xFLAG-sso1Δ1–146 (ii) with sso2Δ. In both cases, harbouring 5xFLAG version of Sso1 and sso2Δ was lethal, which indicates that addition of 5xFLAG disturbed Sso1 function. Individual deletions of SSO1 and SSO2 are alive, but inactivation of both t-SNARE Sso proteins is lethal., pbio.3002263.s009.pdf, Peer reviewed

(A) Our findings suggest that TORC1 blocks Cbk1 kinase activity in the absence of Cdc15 function while cells are arrested in late anaphase at 37 °C, which would promote a defect in the following cell separation as secretory vesicle transporting hydrolases would be unable to fuse into the plasma membrane. (B) Rapamycin inhibits TORC1 and would induce the accumulation of a hypophosphorylated version of Cbk1 that would be able to phosphorylate its key substrates as the exocyct component Sec3, which regulates the function of the SNARE complex at the site of division to promote fusion of secretory vesicle at the plasma membrane. Hydrolases contained in secretory vesicle are released to promote cell separation., Peer reviewed