Resultados de investigación

Circulating antibody-secreting cells are present in the peripheral blood of healthy individuals reflecting the continued activity of the humoral immune system. Antibody-secreting cells typically express CD27. Here we describe and characterize a small population of antibody-secreting class switched CD19+CD43+ B cells that lack expression of CD27 in the peripheral blood of healthy subjects. In this study, we characterized CD27-CD43+ cells. We demonstrate that class-switched CD27-CD43+ B cells possess characteristics of conventional plasmablasts as they spontaneously secrete antibodies, are morphologically similar to antibody-secreting cells, show downregulation of B cell differentiation markers, and have a gene expression profile related to conventional plasmablasts. Despite these similarities, we observed differences in IgA and IgG subclass distribution, expression of homing markers, replication history, frequency of somatic hypermutation, immunoglobulin repertoire, gene expression related to Toll-like receptors, cytokines, and cytokine receptors, and antibody response to vaccination. Their frequency is altered in immune-mediated disorders., Peer reviewed

Increased mitotic activity is associated with the genesis and aggressiveness of many cancers. To assess the clinical value of mitotic activity as prognostic biomarker, we performed a pan-cancer study on the mitotic network activity index (MNAI) constructed based on 54-gene mitotic apparatus network. Our pan-cancer assessment on TCGA (33 tumor types, 10,061 patients) and validation on other publicly available cohorts (23 tumor types, 9,209 patients) confirmed the significant association of MNAI with overall survival, progression-free survival, and other prognostic endpoints in multiple cancer types, including lower-grade gliomas (LGG), breast invasive carcinoma (BRCA), as well as many others. We also showed significant association between MNAI and genetic instability, which provides a biological explanation of its prognostic impact at pan-cancer landscape. Our association analysis revealed that patients with high MNAI benefitted more from anti-PD-1 and Anti-CTLA-4 treatment. In addition, we demonstrated that multimodal integration of MNAI and the AI-empowered Cellular Morphometric Subtypes (CMS) significantly improved the predictive power of prognosis compared to using MNAI and CMS alone. Our results suggest that MNAI can be used as a potential prognostic biomarker for different tumor types toward different clinical endpoints, and multimodal integration of MNAI and CMS exceeds individual biomarker for precision prognosis., Peer reviewed

Additional file 1: Table S1. Summary statistics of the Baikal 1600 m long-read sequencing and metagenomic assembly. Fig. S1. A Principal component analysis (PCA) between deep Lake Baikal metagenomes based on a Bray-Curtis similarity k-mer profile frequencies of sequencing reads. Red and blue dots represent summer and winter Illumina metagenomes, respectively, while the green dot is the sample retrieved in this study and sequenced with PacBio Sequel II. B Phylum-level composition based on 16S rRNA gene fragments (Illumina and PacBio CCS5 reads) of the different metagenomes. The single metagenome highlighted in green corresponds to the PacBio sequencing, whilst the rest of datasets belong to previous Illumina sequencing. The phylum Proteobacteria was divided into its class-level classification. Only those groups with abundance values larger than 1% in any of the metagenomes are shown. C Classification of the 1600 m PacBio CCS5 16S rRNA reads at a higher taxonomic resolution. Only sequences larger than 1000 nucleotides were considered. Sequences ascribed to the Ca. Patescibacteria phylum are highlighted in green. Table S2. Genomic parameters of LAGs recovered in this study. Table S3. Genomic parameters of LAGs recovered in this study with ANI > 99.5% to MAGs retrieved from Lake Baikal 1250 and 1350 m deep. Fig. S2. Alignment of two LAGs that are complete in a single contig and the respective MAG from the Illumina assembly. Table S4. Genomic parameters of the resulting bins from the Baikal 1600 m CCS sequences. The four Baikalibacteria bins are highlighted in yellow. Fig. S3. A Maximum likelihood phylogenetic tree of the Baikalibacteria 16S rRNA genes. Sequences outside the deep branch coming from Figure 1 were used as an outgroup for the tree. The reads from the four read bins are colored in the figure. B Diversity of 16S rRNA sequences of Baikalibacteria bins. Linear representation of selected CCS5 reads (indicated with a red circle in the left panel) containing a 16S rRNA gene. A pairwise blastn comparison among reads was performed to detect orthologous genes. Fig. S4. A Average nucleotide identity based on metagenomic reads (ANIr) of LAGs and the four Baikalibacteria Bins. B ANIr of ten randomly selected sequences of each Baikalibacteria bin. Fig. S5. Metagenomic recruitment of the largest fragment of Baikalibacteria RBin09 on Lake Thun 180 m deep. Fig. S6. Maximum likelihood phylogenetic tree of the a phytoene elongase (LyeJ), b carotenoid 3,4-desaturase (CrtD), and c bisanhydrobacterioruberin hydratase (CruF) proteins., Peer reviewed

(A) cbk1-5E-E164S cbk1-aid cdc15-2 (YMF3905), (B) cbk1-5E-E251S cbk1-aid cdc15-2 (YMF3910), (C) cbk1-5E-E409S cbk1-aid cdc15-2 (YMF3995), (D) cbk1-5E-E615T cbk1-aid cdc15-2 (YMF3906), and (E) cbk1-5E-E711S cbk1-aid cdc15-2 (YMF3907) cells were grown in YPD and arrested in late anaphase by shifting the temperature to 37 °C before the addition of rapamycin to half of the culture for 20 min. 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 determine cell-cycle progression by flow cytometry. (F) 3D Cbk1 structure (PDB 4LQS [48]) in which different protein domains are highlighted. Residue T574 in the activation loop, together with residues D475 and S409 in the kinase domain are denoted. (G) cbk1-T574A cbk1-aid cdc15-2 cells (YMF4191) were grown as above. FACS graphs can be found in the supplementary FACS file (S1 File)., pbio.3002263.s005.pdf, Peer reviewed

Code and data for paper "Discovery of senolytics using machine learning" by Smer-Barreto et al, 2023. Files contain the datasets employed for model training and computational screening, as well as a jupyter notebook with code for model training, feature selection and screening., Peer reviewed

- Document S1. Figures S1–S7. - Table S1. List of differentially expressed genes in WT vs. mutant samples at 12 dpa with statistical data and TPM for each sample, related to Figure 6C. - Table S2. List of differentially expressed genes in WT vs. mutant samples at 14 dpa with statistical data and TPM for each sample, related to Figure 6C. - Table S3. Overlapping genes from the intersection of 12 dpa and 14 dpa lists of differentially expressed genes with statistical data and TPM for each sample, related to Figure 6C. - Table S4. Functional annotation results from common DEGs at 12 and 14 dpa that were upregulated in the WT blastema, related to Figure 6D. - Table S5. Functional annotation results from common DEGs at 12 and 14 dpa that were upregulated in the mutant blastema, related to Figure 6E. - Document S2. Article plus supplemental information., Peer reviewed

DNA extraction protocol (Cobas® DNA Sample Preparation Kit) Table S1. Panel of 29 genes analyzed with next generation sequencing, and distinction between nephropathic and phenocopy genes Table S2. Total variants, silent and non-silent mutations in FSGS patients Table S3. List of variants based on the UnifiedGenotyper and ACMG score Tabla S4. Frequency of variants annotated found in our study ..., Peer reviewed

The globally expanding threat of antibiotic resistance calls for the development of new strategies for abating Gram-negative bacterial infections. The use of extracorporeal blood cleansing devices with affinity sorbents to selectively capture bacterial lipopolysaccharide (LPS), which is the major constituent of Gram-negative bacterial outer membranes and the responsible agent for eliciting an exacerbated innate immune response in the host during infection, has received outstanding interest. For that purpose, molecules that bind tightly to LPS are required to functionalize the affinity sorbents. Particularly, anti-LPS factors (ALFs) are promising LPS-sequestrating molecules. Hence, in this work, molecular dynamics (MD) simulations are used to investigate the interaction mechanism and binding pose of the ALF isoform 3 from Penaeus monodon (ALFPm3), which is referred to as “AL3” for the sake of simplicity, and lipid A (LA, the component of LPS that represents its endotoxic principle). We concluded that hydrophobic interactions are responsible for AL3–LA binding and that LA binds to AL3 within the protein cavity, where it buries its aliphatic tails, whereas the negatively charged phosphate groups are exposed to the medium. AL3 residues that are key for its interaction with LA were identified, and their conservation in other ALFs (specifically Lys39 and Tyr49) was also analyzed. Additionally, based on the MD-derived results, we provide a picture of the possible AL3–LA interaction mechanism. Finally, an in vitro validation of the in silico predictions was performed. Overall, the insights gained from this work can guide the design of novel therapeutics for treating sepsis, since they may be significantly valuable for designing LPS-sequestrating molecules that could functionalize affinity sorbents to be used for extracorporeal blood detoxification., Peer reviewed

Pf of X. laevis oocytes injected with water (W, controls), or expressing non-tagged or Flag-tagged Atlantic halibut HhAqp1ab1-WT or sole SsAqp1ab2-WT. Oocytes injected with SsAqp1ab2-WT were co-injected with non-tagged halibut YwhazLb and exposed to FSK. The data are the mean ± SEM (n = 12 oocytes per treatment, indicated with dots above each bar)., With the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)., Peer reviewed

Histological sections were incubated with Aqp1ab2-Nt and Aqp1ab2_v1 antisera preabsorbed with the immunizing peptides, indicating the specificity of the antibodies. Abbreviations: o, oocyte; y, yolk globule; gv, germinal vesicle; ve, vitelline envelope; cp, capillary; ca, cortical alveoli. Scale bars, 10 μm., With the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)., Peer reviewed