BUSCADOR RECOLECTA

[Introduction]: Drought-associated tree mortality has been increasing worldwide since the last decades, impacting structure and functioning of forest ecosystems, with implications for energy, carbon and water fluxes. However, the understanding of the individual vulnerability to drought-induced mortality is still limited., [Methods]: We aimed to identify the factors that triggered the mortality of the widely distributed Pinus sylvestris L. in an extensive forest area in central Spain. We compared radial growth patterns in pairs of alive and recently dead individuals that co-occur in close proximity and present similar age and size, thereby isolating the effects of size and environment from the mortality process. Temporal dynamics of growth, growth synchrony, and growth sensitivity to water availability (precipitation minus potential evapotranspiration) were compared between alive and recently dead trees., [Results and discussion]: Over the last 50 years, although we did not detect significant differences in growth between alive and dead trees, an increase in the growth synchrony and sensitivity to water availability (i.e. slope of the climatic water balance in the growth model) was observed in all trees as drought intensity increased. 20 years before mortality, dead individuals showed lower growth synchrony and growth sensitivity to water availability than alive ones, without significant differences in growth. Recorded reduction in growth synchrony and growth sensitivity to water availability in dead trees suggests a decoupling between tree growth and climate, which could increase the risk of hydraulic failure and/or carbon starvation under increasingly arid conditions. Thus, the use of reduced growth sensitivity to water availability as potential early-warning signal of tree mortality, together with reduced growth synchrony, should be further explored, particularly in pine species in seasonally dry areas., Peer reviewed

1 file, Supporting information for the article https://doi.org/10.1029/2019GB006229, Text S1 to S6.-- Figures S1 to S16, Peer reviewed

[Introduction]: Refractory/relapsed pediatric acute leukemia are still clinically challenging and new therapeutic strategies are needed. Interactions between Natural Killer Group 2D (NKG2D) receptor, expressed in cytotoxic immune cells, and its ligands (NKG2DL), which are upregulated in leukemic blasts, are important for anti-leukemia immunosurveillance. Nevertheless, leukemia cells may develop immunoescape strategies as NKG2DL shedding and/or downregulation., [Methods]: In this report, we analyzed the anti-leukemia activity of NKG2D chimeric antigen receptor (CAR) redirected memory (CD45RA-) T cells in vitro and in a murine model of T-cell acute lymphoblastic leukemia (T-ALL). We also explored in vitro how soluble NKG2DL (sNKG2DL) affected NKG2D-CAR T cells’ cytotoxicity and the impact of NKG2D-CAR T cells on Jurkat cells gene expression and in vivo functionality., [Results]: In vitro, we found NKG2D-CAR T cells targeted leukemia cells and showed resistance to the immunosuppressive effects exerted by sNKG2DL. In vivo, NKG2D-CAR T cells controlled T cell leukemia burden and increased survival of the treated mice but failed to cure the animals. After CAR T cell treatment, Jurkat cells upregulated genes related to proliferation, survival and stemness, and in vivo, they exhibited functional properties of leukemia initiating cells., [Discussion]: The data here presented suggest, that, in combination with other therapeutic approaches, NKG2D-CAR T cells could be a novel treatment for pediatric T-ALL., Peer reviewed

[Background]: Little is known about whether the overlap syndrome (OS) combining features of chronic obstructive pulmonary disease (COPD) and sleep apnea-hypopnea syndrome increases the risk of stroke associated with COPD itself., [Methods]: We prospectively studied 74 COPD patients and 32 subjects without lung disease. Spirometry and cardiorespiratory polygraphy were used to assess the pulmonary function of the study population and ultrasound measurements of intima media thickness (IMT) as well as the volume of plaques in both carotid arteries were also evaluated., [Results]: Polygraphic criteria of OS were met in 51% of COPD patients. We found that 79% of patients with OS and 50% of COPD patients without OS had atherosclerotic plaques in the left carotid artery (p = 0.0509). Interestingly, the mean volume of atherosclerotic plaques was significantly higher in the left carotid artery of COPD patients with OS (0.07 ± 0.02 ml) than in those without OS (0.04 ± 0.02 ml, p = 0.0305). However, regardless of the presence of OS, no significant differences were observed in both presence and volume of atherosclerotic plaques in the right carotid artery of COPD patients. Adjusted-multivariate linear regression revealed age, current smoking and the apnea/hypopnea index (OR = 4.54, p = 0.012) as independent predictors of left carotid atherosclerotic plaques in COPD patients., [Conclusions]: This study suggests that the presence of OS in COPD patients is associated with larger left carotid atherosclerotic plaques, indicating that OS might be screened in all COPD patients to identify those with higher risk of stroke., Peer reviewed

[Background]: Little is known about whether the overlap syndrome (OS) combining features of chronic obstructive pulmonary disease (COPD) and sleep apnea-hypopnea syndrome increases the risk of stroke associated with COPD itself., [Methods]: We prospectively studied 74 COPD patients and 32 subjects without lung disease. Spirometry and cardiorespiratory polygraphy were used to assess the pulmonary function of the study population and ultrasound measurements of intima media thickness (IMT) as well as the volume of plaques in both carotid arteries were also evaluated., [Results]: Polygraphic criteria of OS were met in 51% of COPD patients. We found that 79% of patients with OS and 50% of COPD patients without OS had atherosclerotic plaques in the left carotid artery (p = 0.0509). Interestingly, the mean volume of atherosclerotic plaques was significantly higher in the left carotid artery of COPD patients with OS (0.07 ± 0.02 ml) than in those without OS (0.04 ± 0.02 ml, p = 0.0305). However, regardless of the presence of OS, no significant differences were observed in both presence and volume of atherosclerotic plaques in the right carotid artery of COPD patients. Adjusted-multivariate linear regression revealed age, current smoking and the apnea/hypopnea index (OR = 4.54, p = 0.012) as independent predictors of left carotid atherosclerotic plaques in COPD patients., [Conclusions]: This study suggests that the presence of OS in COPD patients is associated with larger left carotid atherosclerotic plaques, indicating that OS might be screened in all COPD patients to identify those with higher risk of stroke., Peer reviewed

Additional file 1: Supplementary Figure S1. Expression of RMPs in prostate tumours. A) mRNA expression of RMPs is significantly altered in human normal prostate tissue (N), primary (PT), and metastatic tumour (M). Y-axis shows log2-normalised expression data from Grasso et al. [7],Taylor et al. [6], and Varambally et al. [42] datasets (left panels). Kaplan–Meier curves showing disease-free survival (DFS) of patient groups selected according to decile expression of the indicated RMP genes (middle panel). The right panel shows differential expression in disease-free (DF) and recurrent patient samples from TCGA dataset [3]. B, C) Kaplan–Meier curves representing the disease-free survival (DFS) of patient groups selected according to the decile expression, data from the TCGA dataset [6] (TCGA: n = 491). D, E) PCa cell lines show increased METTL1 protein and mRNA levels. Western blotting showing METTL1, WDR4, AR, PTEN and phosphor-S6K expression (D), and qPCR (E) of normalised mRNA expression (lower panel) in prostate epithelial cell lines (PWR-1E and RWPE-1), benign prostatic hyperplasia cell lines (BPH1), and prostate cancer cell lines (VCap, C4-2, LNCaP, 22Rv1, DU145, and PC3). Mean ± SD (D, E). Statistical tests: ANOVA test (A), log-rank Cox test (A, B); Student’s t-test with Welch’s correction, grouping all data from benign vs. all data from tumour cells ****p < 0.0001 (E)., Consejo Superior de Investigaciones Cientificas (CSIC), Peer reviewed

Additional file 2: Supplementary Figure S2. Regulation of METTL1 expression in PCa. A) Correlation analysis between METTL1 and KLK3 expression, and METTL1 and AR expression in the human primary PCa expression datasets. The plotted values correspond to the log2-normalised expression values. Black line represents linear regression, grey area indicates the limits of the confidence intervals. Pearson's correlation coefficient (R) and p-values are indicated. Grasso n = 88; Taylor n = 183; TCGA n = 497. B) No effect on mRNA expression levels of METTL1 upon dihydrotestosterone (DHT) treatment in LNCaP cells. Mean ± SD, n = 3. C) Graphical representation of PI3K inhibition by different compounds. Small molecule inhibitors used: pan-PI3K inhibitor BKM-120 (BKM), AKT inhibitor MK2206 (MK), mTORC1 inhibitor rapamycin (RAPA), and mTORC1/2 inhibitor Torin (TOR). D, E) METTL1 expression is affected by the inhibition of downstream PI3K pathway members. Western blotting (D) and RT-qPCR (E) analyses of METTL1 expression upon PI3K pathway inhibition in PC3 cells. The cells were treated for 48 h (D) or 8 h (E). Mean ± SD, n = 2 (D) and n = 6 (E). F) Representative images of immune stained sections for Mettl1 and markers for luminal (AR), and basal (K14) cells in Pten-KO prostates (dorsal and ventral lobes) in invasive carcinoma (6 months old mice) and in aged-match wild-type (WT) prostates. Scale bars represent 50 μm. Statistical tests: One-tailed Student’s t-test (B, D, E). *p < 0.05, **p < 0.01, ***p < 0.001., Consejo Superior de Investigaciones Cientificas (CSIC), Peer reviewed

Additional file 3: Supplementary Figure S3. tRNAs are methylated at guanine-7 by METTL1. A) Western blot showing the expression of doxycycline-inducible HA-METTL1 in PC3 cells. B, C) Density distribution of reads per million (RPM) identified after PAR-CLIP analysis of METTL1-bound tRNAs (B) and other RNA species (C) in PC3 (control) and HA-METTL1 expressing PC3 cells. The red line indicates the third lower quartile of total RPMs. D) tRNAs are the most common RNA species that bind Flag-METTL1 in HEK293T cells. Density distribution of reads per million (RPM) identified after PAR-CLIP analysis of METTL1-bound RNAs: The red line indicates the third lower quartile of total RPMs. Data were retrieved from Bao et al. study. E) tRNAs comprise half of the reads of all RNAs bound to METTL1 after PAR-CLIP analysis in HEK293 cells. F) Boxplot representing the median with interquartile range of reads per million (RPM) per transcript bound to METTL1 in HEK293T cells. G) Schematic representation of genomic editing introduced by CRISPR-Cas9 technology in the PC3 METTL1 KO clones used in this study. H) METTL1 mRNA expression levels in METTL1 KO, and WT control clones used in this study. Mean ± SD, n = 3. I) Graphics representing the experimental workflow followed to generate AlkAniline-seq libraries. J) Normalised cleavage signals for METTL1-methylated tRNAs Cys and Ile, and METTL1 non-methylated tRNAs His and Glu in PC3 WT and METTL1 KO cells., Consejo Superior de Investigaciones Cientificas (CSIC), Peer reviewed

Additional file 4: Supplementary Figure S4. Accumulation of 5'tRNA fragments in METTL1 KO cells. A) The most common sequences of 5' terminal guanines (5G) containing 5'TOGs and 4G 5'TOGs are shown. B, C) Northern blot showing the absence of 5'tRNA fragments derived from Lys and Pro tRNA in PC3 WT METTL1 KO cells under normal conditions. D) qPCR showing reduced expression of METTL1 in 22Rv1 cells upon METTL1 silencing using siRNA. Mean ± SD, n = 3. E) Dot blot against m7G showing reduced methylation levels in tRNAs extracted from METTL1-silenced 22Rv1 cells. Mean ± SD, n = 3. F) Northern blot detection of Cys-derived 5'TOGs in METTL1-silenced 22Rv1 cells. Mean ± SD, n = 3. G) Northern blot detection of Cys-derived 5'TOGs in PC3 METTL1 KO cells (second biological replicate), unexposed (0 h) or after 2 and 8 h of oxidative stress exposure. H) Northern blot detection of Lys-derived 5'TOGs in PC3 METTL1 KO cells unexposed (0 h) or after 2 and 8 h of oxidative stress exposure. I) Western blotting of DU145 METTL1 KO and WT cell clones generated using CRISPR-Cas9 technology and parental DU145 (DU) cells. J) Northern blot detection of Cys-derived 5'TOGs in DU145 METTL1 KO cells (second biological replicate), unexposed (0 h) or after 2 and 8 h of oxidative stress exposure. The loading control by red-safe staining (tRNA) is shown in the bottom panels (B, C, F–H, J). Bands corresponding to full-length tRNAs are indicated with stars, and 5’tRNA fragments are indicated with arrows. Statistical tests: One-tailed Student’s t-test (C-F). *p < 0.05., Consejo Superior de Investigaciones Cientificas (CSIC), Peer reviewed

1 file, The current trend of climatic alterations will accelerate the modification of the ocean system by, among other aspects, changing the metal speciation and its bioavailability which may have an impact in their accumulation by marine organisms. Understanding the impact of these potential changes is essential for future risk assessment of metal contamination. In the present study, we selected the species Mediterranean mussel (Mytilus galloprovincialis) as the main European aquaculture production bivalve and due to its widespread use for biomonitoring purposes. A long-term test (2 months) was carried out to explore the impact that global change in the marine environment (warming and CO2 increase) may exert on the accumulation of dissolved trace metals (Cu, Co, Pb, Cd, Cr, As and Ni) in different body parts of mussels (foot and soft tissue). Studied mussels were collected at two different climatic locations (Atlantic and Mediterranean Sea) and exposed to unspiked, unpolluted seawater from the Vigo Ria (NW Iberian Peninsula). Results showed that under the global change conditions proposed in this study (1100 pCO2 and 25 °C), the increase in temperature resulted in a lower condition index and byssus strength for mussels from Atlantic Sea, while Mediterranean sea mussels, adapted to higher temperatures, did not show remarkable variations. According to trace metals accumulation in different body parts of the studied mussels, it was observed that the effect of increasing CO2 alone did not show to have an impact in the bioaccumulation, but the combined stressors (increase in CO2 and temperature) may lead to an increase in the bioaccumulation for some elements. The increase in temperature resulted in a decrease of the Cu content of foot tissue (byssus gland) in mussels from Atlantic Sea, which is in accordance with the lower byssus strength observed under such conditions. Our results indicate that the expected seawater temperature increase, which will be produced gradually during next decades, should be further study to ensure the species adaptability and aquaculture production, Peer reviewed