BUSCADOR RECOLECTA

Peer reviewed

[Objectives] Inflammatory biomarkers, as indicators of biological states, provide a valuable approach for accurate and reproducible measurements, crucial for the effective management of mild traumatic brain injury (mTBI) in pediatric patients. This study aims to assess the diagnostic utility of blood-based inflammatory markers IL6, IL8, and IL10 in children with mTBI, including those who did not undergo computed tomography (CT) scans., [Methods] A prospective multicentric cohort study involving 285 pediatric mTBI patients was conducted, stratified into CT-scanned and non-CT-scanned groups within 24 h post-trauma, alongside 74 control subjects. Biomarker levels were quantitatively analyzed using ELISA. Sensitivity and specificity metrics were calculated to determine the diagnostic efficacy of each biomarker., [Results] A total of 223 mTBI patients (78%) did not undergo CT scan examination but were kept in observation for symptoms monitoring at the emergency department (ED) for more than 6 h (in-hospital-observation patients). Among CT-scanned patients (n = 62), 14 (23%) were positive (CT+). Elevated levels of IL6 and IL10 were found in mTBI children compared to controls. Within mTBI patients, IL6 was significantly increased in CT+ patients compared to both CT– and in-hospital-observation patients. No significant differences were observed for IL8 among the compared groups. IL6 yielded a specificity of 48% in identifying CT– and in-hospital-observation patients, with 100% sensitivity in excluding all CT+ cases. These performances were maintained whether IL6 was measured within 6 h or within 24 h after the trauma., [Conclusion] The inflammatory marker IL6 emerges as a robust biomarker, showing promising stratification value for pediatric mTBI patients undergoing CT scans or staying in observation in a pediatric ED., Peer reviewed

Technical details for the experimental setups used to characterize the physicochemical properties of ZEO-1, along with the computational details for the DFT modeling; PXRD, nitrogen adsorption/desorption, TGA figures, and SEM images for different ZEO-1 samples synthesized under different conditions; 29Si, 27Al (1D and MQMAS), 31P, and 1H solid-state NMR results; in situ IR spectroscopy results illustrating the determination of the molar extinction coefficient of the 1545 cm–1 pyridinium band, comparison between the protonic form of ZEO-1 and its Cs form, and the series of spectra recorded after sending discrete monodoses of CO into the IR cell; DFT modeling figures presenting the relative stability of the ZEO-1 structure, the calculated aluminum distribution based on the Boltzmann distribution, and the relative deprotonation energy of all the bridging hydroxyl groups versus the Al positions; catalytic test results of the initial yields of the principal cracking products as a function of initial n-hexane conversion and the n-hexane conversion as a function of TOS for ZEO-1, Beta, and USY; T–O–T bond angles corresponding to the aluminum substitution for each T-atom within the ZEO-1 framework and its corresponding isotropic chemical shift; and relative stability of the modeled 21 anionic structures and 84 neutral structures with different locations of Al and of the charge-compensating proton around them., Peer reviewed

HERFD-XANES spectra taken at the maximum of Ru Lα1 emission line for the whole Ca3Mn2–xRuxO7 (0.05 ≤ x ≤ 0.9) samples at room temperature, k2-weighted EXAFS oscillations in k-space as a function of Ru concentration (x) at Mn K and Ru K edges, and detailed EXAFS analysis for the Ca3Mn2–xRuxO7 (0.1 ≤ x ≤ 0.7) series at the Mn and Ru K-edges., Peer reviewed

Sample description, electron microscopy images and analysis, XRD of time and temperature series, TEM and SEM images of annealed samples, FIB cross sections, and schematic of synchrotron experiment., Peer reviewed

Figure S1. Morphological, magnetic and compositional analysis of the elaborated MNPs. (a) quasi-spherical shape and size of 21 ± 4 nm according to TEM images; (b) hysteresis loop showing saturation magnetization of 54 emu/g and 12 Oe of coercive field; and (c) EDS analysis showing Fe2.9Mn0.1O4 stoichiometry; Figure S2. Characterization of MSNs. (a) typical TEM image from elaborated MSNs; (b) size of 33.3 ± 4.2 nm; (c) FTIR spectrum; and (d) N2 adsorption isotherm for BET analysis; Figure S3. ATR-FTIR spectra of unloaded (black), curcumin-loaded (blue) and ketorolac-loaded (red) MSNs with the characteristic silica bands marked in black and two bands characteristic for ketorolac tromethamine at 2970 cm−1 and 2360 cm−1 marked in red. Most of the loaded drug is inside the pores and thus not visible in ATR-FTIR; Figure S4: Cumulative drug release of curcumin from MSNs at pH 7.4 and 5.5 and 37 °C. Curcumin shows a higher release at more acidic pH, with a maximum of 2.2% after 12 days; Figure S5: Cumulative drug release of ketorolac tromethamine from MSNs at pH 7.4 and 5.5 and 37 °C. Ketorolac showed a higher release at pH 7.4, up to 11.4% after 12 days, compared to 8.7% at pH 5.5; Figure S6. Contact angle measurement of CurPAN. With a contact angle of 143.6°, the curcumin-loaded fibers are very hydrophobic; Figure S7. Images from electrospun mats, (a) ENFs-KET, (b) ENFs-@MSNs-KET, (c) ENFs-CUR, (d) ENFs-@MSNs-CUR., Peer reviewed

X-ray data (Tables S1, S2 and S3); representation of zwitterionic en2ampy (Scheme S1); representation of the planar [Cu(en2ampy)]2+ unit (Figure S1); representations of the crystal packing of 1 and 2 (Figures S2–S5); [DNA]/(ϵa-ϵf) vs. [DNA] plot for 1 (Figure S6); UV-Vis spectra of 2 (Figure S7); [DNA]/(ϵa-ϵf) vs. [DNA] plot for 2 (Figure S8); Stern-Volmer plots for 1 (Figure S9); Competitive displacement assays for DNA-EB adducts for 2 (Figure S10); Stern-Volmer plots for 2 (Figure S11); Plot of relative viscosity for 2 (Figure S12); gel electrophoresis image for 2 (Figure S13); Percent cell viability versus complex concentration for 1 and 2 (Figure S14); IR and NMR spectra of en2ampy (Figures S15–S17); IR spectra of 1 and 2 (Figures S18 and S19., Peer reviewed

Throughout the family of coronaviruses, structured RNA elements within the 5′ region of the genome are highly conserved. The fifth stem-loop element from SARS-CoV-2 (5_SL5) represents an example of an RNA structural element, repeatedly occurring in coronaviruses. It contains a conserved, repetitive fold within its substructures SL5a and SL5b. We herein report the detailed characterization of the structure and dynamics of elements SL5a and SL5b that are located immediately upstream of the SARS-CoV-2 ORF1a/b start codon. Exploiting the unique ability of solution NMR methods, we show that the structures of both apical loops are modulated by structural differences in the remote parts located in their stem regions. We further integrated our high-resolution models of SL5a/b into the context of full-length 5_SL5 structures by combining different structural biology methods. Finally, we evaluated the impact of the two most common VoC mutations within 5_SL5 with respect to individual base-pair stability., Peer reviewed

S1 Fig. Ccr4 and Rnr4 help to tolerate MMS and HU sensitivity. The absence of Ccr4 and Rnr4 causes MMS and HU sensitivity as determined by ten-fold serial dilutions at the indicated concentrations. The experiment was repeated twice with similar results. https://doi.org/10.1371/journal.pgen.1011148.s001 (TIFF) S2 Fig. Characterization of the MCM/RNR interactions. (A) HU promotes MCM/RNR interactions, as determined by CoIP and western blot in asynchronous cultures treated with 0.2M HU for 2 hours. Untreated and treated cells with 0.025% MMS for 2 hours were included as control. (B) Mcm7 interacts with Rnr1, as determined by CoIP and western blot in asynchronous cultures of wild-type cells. This interaction is lost in the MCM-VC mutant. Asterisks represent unspecific bands. https://doi.org/10.1371/journal.pgen.1011148.s002 (TIFF) S3 Fig. Subcellular location and intensity of the Mcm4-VC/Rnr4-VN interaction. (A-C) BiFC analysis of the subcellular location and intensity of the Mcm4VC/Rnr4-VN interaction during the cell cycle in cells synchronized in G1 and released into fresh medium in the absence and presence of 0.025% MMS. (A) Representative images of dividing cells expressing Mcm4-VC and/or Rnr4-VN. (B) Intensity of the Mcm4VC/Rnr4-VN fluorescence signal in the nucleus and cytoplasm of cells grouped according to the bud-to-mother size ratio to compare similar stages of DNA replication with and without DNA damage. The Venus signal was calculated in equivalent areas of the nucleus and the cytoplasm. The SuperPlot shows the mean and SEM from three independent experiment (represented by color dots). (C) Intensity of the Mcm4-VC/Rnr4-VN fluorescence signal as determined by cell sorting analysis. Cells were grouped in G1, early and late S phase (eS and lS) and G2/M according to the presence and size of the bud. The amount of the MCM and Rnr2/Rnr4 complexes was followed by quantifying the GFP signal in cells expressing Mcm4-GFP and Rnr4-GFP under the same experimental conditions. The mean and SEM of 4–7 fluctuations tests are shown. https://doi.org/10.1371/journal.pgen.1011148.s003 (TIFF) S4 Fig. Supplementary information related to Fig 4. (A) The expression of Mcm4-VC or Rnr4-VN causes HU and MMS sensitivity as determined by ten-fold serial dilutions at the indicated concentrations (top panels). The MCM4-VC allele is recessive for HU and MMS sensitivity, as determined in cells transformed with a centromeric plasmid expressing Mcm4 from its own promoter (p316MCM4) or an empty vector (pRS316) (bottom panels). The experiments were repeated twice with similar results. (B) Levels of dNTPs in RNR4-VN and wild-type cells transformed or not with plasmid p314RNR4. The mean and SEM of four independent experiments are shown. (C-D) Frequency of petite formation in the indicated strains. The mean and SEM of 3 fluctuations tests are shown. (E) The HU sensitivity of the RNR4-VN mutant can be complemented with a Rnr4 expressing plasmid as determined by ten-fold serial dilutions at the indicated concentrations. The experiments were repeated twice with similar results. (F-G) Analysis of the MCM/RNR interactions in cells expressing Mcm4-VC and/or Rnr4-VN transformed with plasmid p314RNR4. Effect of tagging Mcm4 and Rnr4 with VC and VN, respectively, on the physical interactions between the MCM and RNR complexes in cells transformed with plasmid p314RNR4, as determined by CoIP and western blot in asynchronous cultures treated (F) or not (G) with 0.025% MMS for 2 hours. Asterisks indicate unspecific bands. Rnr4-VN* indicates a slow-migrating Rnr4-VN form. An over-exposition (OE) of the Rnr4 IP signals is shown in the bottom panel. CoIP analyses were performed twice with similar results. https://doi.org/10.1371/journal.pgen.1011148.s004 (TIFF) S5 Fig. The Mcm4-VC chimera leads to DNA damage accumulation. (A-B) Rad53 phosphorylation (A) and Sml1 levels (B) at asynchronous cultures of the indicated strains transformed with plasmid p314RNR4 in the absence of genotoxic agents. A wild-type culture incubated with 0.025% MMS for 1 hour was included as positive control. (C) Spontaneous accumulation of Rad52 foci at asynchronous cultures of the indicated strains transformed with plasmid p314RNR4. (D) Cell cycle progression of MCM-VC and wild-type strains synchronized in G1 and released into S phase, as determined by cell sorting analysis. (E) The MCM4-VC allele is recessive for the accumulation of spontaneous and HO-induced Rad52 foci, as determined in cells transformed with a centromeric plasmid expressing Mcm4 from its own promoter (p316MCM4) or an empty vector (pRS316). The kinetics of HO-induced Rad52 foci formation and resolution was performed as indicated in Fig 5B. (C, E) The mean and SEM of three independent experiments are shown. Asterisks indicate statistically significant differences according to an unpaired two-tailed Student’s t-test (three asterisks represent a P-values <0.001). https://doi.org/10.1371/journal.pgen.1011148.s005 (TIFF) S6 Fig. Characterization of checkpoint and mutagenesis in MCM4-VC cells. (A) Checkpoint activation is proficient in MCM4-VC mutants. Checkpoint activation in MCM4-VC and RNR4-VN mutants, as determined by western blot against Rad53 in asynchronous cultures treated with 0.2M HU for 60 minutes. Cell cycle progression was determined by cell sorting analysis. The P-Rad53/Rad53 ratio was determined by dividing the upper bands signal (Rad53-P) by the lower band signal (Rad53). Experiments were performed with cells transformed with plasmid p314RNR4. (B) The MCM4-VC allele is recessive for hypermutagenesis, as determined in MCM4-VC and wild-type cells transformed with a centromeric plasmid expressing Mcm4 from its own promoter (p316MCM4) or an empty vector (pRS316). The frequency of mutagenesis was determined from colonies grown with 0.0075% MMS or 50mM HU. The mean and SEM of three independent experiments are shown. Asterisks indicate statistically significant differences according to an unpaired two-tailed Student’s t-test (three asterisks represent a P-values <0.001). https://doi.org/10.1371/journal.pgen.1011148.s006 (TIFF) S7 Fig. dNTP levels in MCM4-VC and RNR4-VN mutants. (A) Levels of dNTPs in MCM4-VC and RNR4-VN mutants in the absence and presence of 0.05% MMS, as determined in mid-log phase asynchronous cultures of the indicated strains transformed with plasmid p314RNR4. The mean and SEM of four independent experiments are shown. Asterisks (relative to the wild type) and dots (between the indicated values) show statistically significant differences according to an unpaired two-tailed Student’s t-test (one, two and three asterisks represent P-values <0.05, <0.01 and <0.001, respectively). (B) The amount of Rnr3 is increased in MCM4-VC cells, as determined by western blot analysis from asynchronous cultures of MCM4-VC and wild-type cells. The amount of Rnr3 was normalized to the amount of Pgk1. The mean and SEM of three independent experiments are shown. Asterisks indicate statistically significant differences according to an unpaired two-tailed Student’s t-test (two asterisks represent a P-values <0.01). https://doi.org/10.1371/journal.pgen.1011148.s007 (TIFF) S8 Fig. MMS-induced uSCE in the MCM4-VC mutant is independent of Rev1. The frequency of HR was determined from colonies grown in the presence of 0.0075% MMS. The mean and SEM of 4 fluctuations tests are shown. https://doi.org/10.1371/journal.pgen.1011148.s008 (TIFF) S9 Fig. Raw data for Figure panels. Original blots for the indicated figure panels are shown. https://doi.org/10.1371/journal.pgen.1011148.s009 (PDF) S1 Table. Saccharomyces cerevisiae strains used in this study. Strains, genotypes, references and Figures panels where they have been used are indicated. https://doi.org/10.1371/journal.pgen.1011148.s010 (DOCX) S2 Table. Oligonucleotydes used in this study. DNA sequences and Figures panels where they have been used are indicated. https://doi.org/10.1371/journal.pgen.1011148.s011 (DOCX) S3 Table. Raw data for Figure plots. Raw values to build graphs in the indicated Figure panels are shown. https://doi.org/10.1371/journal.pgen.1011148.s012 (XLSX), Peer reviewed

"Circular dichroism The differential transmittance measurements were carried out in a custom-made optical setup. A white-light tungsten halogen lamp (Ocean Optics, HL-2000-HP, FL, USA) is coupled to a silver reflective collimator (RC08SMA-P01, Thorlabs) and used as an excitation light source. The light beam is passed through a Glan-Thompson Calcite Polarizer (GTH10M, Thorlabs) and directed to a super achromatic quarter wave-plate (SAQWP05M-700, Thorlabs) oriented at ±π/4 compared to the polarization direction on a rotation mount (ELL14, Thorlabs) to obtain a circularly polarized light beam. The optical elements are automatically controlled by custom software (LabView NXG). The sample is positioned at the focal plane of a pair of 4x objectives (RMS4X, NA = 0.1, Olympus). Finally, the transmitted light is fiber-coupled to a spectrometer (Ocean Optics, QEPro-FL). Steady-state circularly polarized photoluminescence Chiral photoluminescence is characterized using the same optical elements as for the dichroic transmittance but reversed in order. First, the emitted chiral photoluminescence is directed to a quarter wave-plate (10RP52-1B, Newport) oriented at ±π/4 and then to a linear polarizer (20LP-VIS-B, Newport), filtering one of the circular components. The excitation source consisted of a 200ps pulsed laser source (LDH-P-C-405 laser driven with a PDL 800B driver with 5–80 MHz repetition rate)) with its wavelength peak at 405nm. In the case of the unpolarized excitation study, the light source used for excitation was an unpolarized LED centered at 405nm (M405L4, Thorlabs) coupled to a 4x (RMS4X, NA=0.1, Olympus) objective used to collimate the diverging emitted beam and refocused with an achromatic 50mm lens upon the metasurface. Time-resolved circularly polarized photoluminescence The optical set-up used for the time-resolved photoluminescence was PicoQuant Time Correlated Single Photon Counting system (Time Harp 260 PICO board, 25 ps temporal resolution; PMA Hybrid 40 detector, 250 ps response time; 405 nm LDH-P-C-405 laser driven with a PDL 800B driver with 5–80 MHz repetition rate) equipped with a compact monochromator (Solar Laser Systems). Photoluminescence lifetimes were retrieved using the PicoQuant FluoFit Pro software, and fitting the PL decay data accounting for instrument response function.", Peer reviewed