Resultados de investigación

Supplementary Table 3 Clinical and biochemical parameters of available patients harboring the c.283+1G>A variant in THRB gene. Abnormal values are in bold numbers., Inherited retinal dystrophies (IRDs) are a clinically and genetically heterogeneous group of disorders that often severely impair vision. Some patients manifest poor central vision as the first symptom due to cone-dysfunction, which is consistent with cone dystrophy (COD), Stargardt disease (STGD), or macular dystrophy (MD) among others. Here, we aimed to identify the genetic cause of autosomal dominant COD in one family. WGS was performed in 3 affected and 1 unaffected individual using the TruSeq Nano DNA library kit and the NovaSeq 6,000 platform (Illumina). Data analysis identified a novel spliceogenic variant (c.283 + 1G>A) in the thyroid hormone receptor beta gene (THRB) as the candidate disease-associated variant. Further genetic analysis revealed the presence of the same heterozygous variant segregating in two additional unrelated dominant pedigrees including 9 affected individuals with a diagnosis of COD (1), STGD (4), MD (3) and unclear phenotype (1). THRB has been previously reported as a causal gene for autosomal dominant and recessive thyroid hormone resistance syndrome beta (RTHβ); however, none of the IRD patients exhibited RTHβ. Genotype-phenotype correlations showed that RTHβ can be caused by both truncating and missense variants, which are mainly located at the 3′ (C-terminal/ligand-binding) region, which is common to both THRB isoforms (TRβ1 and TRβ2). In contrast, the c.283 + 1G>A variant is predicted to disrupt a splice site in the 5′-region of the gene that encodes the N-terminal domain of the TRβ1 isoform protein, leaving the TRβ2 isoform intact, which would explain the phenotypic variability observed between RTHβ and IRD patients. Interestingly, although monochromacy or cone response alterations have already been described in a few RTHβ patients, herein we report the first genetic association between a pathogenic variant in THRB and non-syndromic IRDs. We thereby expand the phenotype of THRB pathogenic variants including COD, STGD, or MD as the main clinical manifestation, which also reflects the extraordinary complexity of retinal functions mediated by the different THRB isoforms., Peer reviewed

Supplementary Figure 2 Generation of a HVEM-deficient A20-GFP tumor cell line using a CRISPR-Cas9 approach. (A) Mouse HVEM gene is encoded by eight exons. Deletion of exon 1 encoding the first part of the signal peptide led to abrogation of HVEM expression on the cell membrane of tumor cells. Different domains of HVEM are abbreviated as follows: TM, transmembrane region; CRD, Cystein-Rich Domain; IC, intracellular domain. (B) Cleavage efficiency of different sgRNA guides designed over exon 1: control HVEM DNA target sequence, HVEM DNA target sequence plus Cas9 or HVEM DNA plus Cas9 incubated with different sgRNA guides were loaded and run in a 1% agarose gel. (C) PCR amplification of HVEM mutation of exon 1 from A20-GFP HVEM WT (LCL-7) tumor cell line and A20-GFP HVEM KO deficient tumor cell line (LCL-25). The expected band for the amplicon obtained with the flanking primers designed on exon 1 was 1090 bp (A20 HVEM WT), whereas in HVEM deficient tumor cell line was 618 bp. The indel mutation introduced into exon 1 encompassed 472 bp affecting the 5´UTR region, exon 1, exon 1 and part of intron 1-2 sequences. (D) Amino acid sequence alignment showing the complete deletion of the first part of the signal peptide encoded by exon 1 of mouse HVEM gene. The alignment of the amino acid sequence of the HVEM WT protein versus HVEM KO protein was performed with Clustal Omega (http://www.ebi.ac.uk/tools/clustalo/. An asterisk displays identical amino acids indicating perfect alignment (*). (E) Flow cytometry histograms of A20 HVEM WT, A20-GFP HVEM WT (LCL-7) and A20-GFP HVEM KO (LCL-25) cell lines stained with biotinylated isotype matched control (rat IgG2a, AFRC MAC 157) antibody or biotinylated rat anti-mouse HVEM monoclonal antibody (clone 6C9) and biotinylated rat anti-mouse BTLA (clon 4G12b) mAbs. The binding reactions were developed using streptavidin (SA)-Alexa Fluor 647 or SA-BV421. (F) In vitro rate of proliferation of HVEM WT and HVEM KO cell lines. Four replicates of A20 HVEM WT or A20 HVEM KO tumor cells were seeded in a 24 well plate at the rate of 5000 cells per well and cell counting was performed every day over a period of 6 days. Total number of cells (×106) in culture is plotted at different time points., Introduction: A high frequency of mutations affecting the gene encoding Herpes Virus Entry Mediator (HVEM, TNFRSF14) is a common clinical finding in a wide variety of human tumors, including those of hematological origin., Methods: We have addressed how HVEM expression on A20 leukemia cells influences tumor survival and its involvement in the modulation of the anti-tumor immune responses in a parental into F1 mouse tumor model of hybrid resistance by knocking-out HVEM expression. HVEM WT or HVEM KO leukemia cells were then injected intravenously into semiallogeneic F1 recipients and the extent of tumor dissemination was evaluated., Results: The loss of HVEM expression on A20 leukemia cells led to a significant increase of lymphoid and myeloid tumor cell infiltration curbing tumor progression. NK cells and to a lesser extent NKT cells and monocytes were the predominant innate populations contributing to the global increase of immune infiltrates in HVEM KO tumors compared to that present in HVEM KO tumors. In the overall increase of the adaptive T cell immune infiltrates, the stem cell-like PD-1- T cells progenitors and the effector T cell populations derived from them were more prominently present than terminally differentiated PD-1+ T cells., Conclusions: These results suggest that the PD-1- T cell subpopulation is likely to be a more relevant contributor to tumor rejection than the PD-1+ T cell subpopulation. These findings highlight the role of co-inhibitory signals delivered by HVEM upon engagement of BTLA on T cells and NK cells, placing HVEM/BTLA interaction in the spotlight as a novel immune checkpoint for the reinforcement of the anti-tumor responses in malignancies of hematopoietic origin., Peer reviewed

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/w15203679/s1, Figure S1: Chemical structure of pollutants, Figure S2: Pollutants structures as a function of the pH; Table S1: Characteristics of the pharmaceuticals; Supplementary references: [93,94,95,96,97,98]., Peer reviewed

Additional SEM double deposition anneal; annealed and etched in HF; histogram Au-NP diameter; automated measurement; annealing different temperatures; annealing in air and vacuum; quadruple deposition 20 nm; scattering and absorption cross section; reflectance spectra substrate thickness; Raman spectra different samples; raw spectra; additional surface maps; Au-NP diameter vs enhancement; laser power Raman study; Raman band table., Peer reviewed

Research conducted in the framework of MIRACLE Project (Photonic Metaconcrete with Infrared RAdiative Cooling capacity for Large Energy savings, GA 964450), coordinated by Dr. Jorge Sánchez Dolado, from Centro de Física de Materiales (CFM)., Dataset that includes the geometric characteristics (7) and the emissivity curve (110 points between 0-25um) of four vertically embedded alumnium fibers in CSH substrate., No

Research conducted in the framework of MIRACLE Project (Photonic Metaconcrete with Infrared RAdiative Cooling capacity for Large Energy savings, GA 964450), coordinated by Dr. Jorge Sánchez Dolado, from Centro de Física de Materiales (CFM)., Cementitious matrices are composed of Portlandite (CH) and a porous element, called C-S-H gel that resembles Tobermorite minerals. The dielectric response of CH has been calculated through Force Field simulations by using a software package called GULP (https://gulp.curtin.edu.au/). This datasets annexes the output of our FF simulation for CH, where the complex dielectric function of this minerals can be found. This dataset was used for the publication https://doi.org/10.1021/acsaom.3c00082., European Commission: 964450, Peer reviewed

Research conducted in the framework of MIRACLE Project (Photonic Metaconcrete with Infrared RAdiative Cooling capacity for Large Energy savings, GA 964450), coordinated by Dr. Jorge Sánchez Dolado, from Centro de Física de Materiales (CFM)., Cementitious matrices are composed of Portlandite (CH) and a porous element, called C-S-H gel that resembles Tobermorite minerals. The dielectric response of Tobermorite has been calculated through Force Field simulations by using a software package called GULP (https://gulp.curtin.edu.au/). This datasets annexes the output of our FF simulation for Tobermorite , where the complex dielectric function of this minerals can be found. This dataset was used for the publication https://doi.org/10.1021/acsaom.3c00082., No

Table S1. Unitary costs (year 2020).-- Table S2. Baseline characteristics of the study population before propensity score matching., [Objective] This study aims to compare the hospitalization rate in individuals with schizophrenia who started their treatment with aripiprazole once monthly (AOM400) or atypical oral antipsychotics (OA) in Spain., [Methods] This is an observational and retrospective study based on the electronic medical records from the BIG-PAC database. The study population consisted of individuals diagnosed with schizophrenia who initiated their treatment with AOM400 (AOM cohort) or atypical OA (OA cohort) from 01/01/2017 to 31/12/2019. A 1:1 propensity score matching (PSM) procedure was conducted to match individuals of both cohorts. The number and duration of hospitalizations, persistence to treatment, healthcare resources use, and costs were analyzed after 12 months., [Results] After the PSM, 1,017 individuals were included in each cohort [age: 41.4 years (SD: 10.6); males: 54.6%]. During the follow-up period, the AOM cohort had a 40% lower risk of hospitalization than the OA group [HR: 0.60 (95% confidence interval, CI: 0.49–0.74)]. The median time to the first hospitalization was longer in individuals with AOM400 compared to those with OA (197 days compared to 174 days; p < 0.004), whereas hospital admissions were shorter (AOM400: 6 compared to OA: 11 days; p < 0.001). After 12 months, individuals receiving AOM400 were more persistent than those with OA (64.9% compared to 53.7%; p < 0.001). The OA cohort required more healthcare resources, mainly visits to primary care physicians, specialists, and emergency rooms than those receiving AOM400 (p ≤ 0.005 in all comparisons). AOM400 reduced the costs of hospitalizations, and emergency room, specialist and primary care visits by 50.4, 36.7, 16.1, and 10.9%, respectively, in comparison to the treatment with atypical OA. AOM400 led to annual cost savings of €1,717.9 per individual, from the societal perspective., [Conclusion] Aripiprazole once monthly reduces the number and duration of hospitalizations, together with the treatment costs of schizophrenia, as it reduces the use of healthcare resources and productivity losses in these individuals., Peer reviewed

Zeta potential of AuNPs and silica-coated AuNPs (Figure S1); low magnification TEM images of different samples of AuNPs (Figure S2); low magnification bright-field TEM images of non-centrifuged AuNPs (Figure S3); photographs of Au@SiO2 NP precipitates and re-dispersions obtained from different centrifuge speeds (Figure S4); size distribution of core diameter and shell thickness (Figure S5); percentage of aggregated AuNPs and single AuNPs present in Au@SiO2 NPs obtained from 2000, 4000, and 6000 rpm centrifugation (Figure S6); TEM images and SERS spectra with 10–5 M Rh6G of Au@SiO2 NPs with thick silica shell (Figure S7); HRTEM image of a single-Au@SiO2 NP (Figure S8); high-magnification TEM images of multi-Au@SiO2 NPs (Figure S9); TEM images showing the heat effect of electron beam of TEM (Figure S10); effect of ionic strength on the NPs (Figure S11); effect of adding Rh6G to the NPs (Figure S12); calculation of SERS enhancement factor (Table S1); FDTD simulation for AuNPs (Figure S13); cyclic voltammetry with different scan rates (Figure S14); reproducibility, long-term stability, repeatability, and selectivity of Au@SiO2_2000/GCE for glucose sensing (Figure S15)., Peer reviewed

Additional file 1: Supplementary Fig. 1. Study design and Flow-chart., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed