Caracterización simbiótica y filogenética de rizobios que nodulan la nueva especie Lupinus mariae-josephi

Una nueva especie de altramuz, Lupinus mariae-josephi, ha sido identificado recientemente en Valencia (Pascual, H.). Esta especie, a diferencia de las descritas en la Península ibérica y en el viejo mundo, no crece en suelos ácidos sino en suelos alcalinos y con alto contenido en calcio. El objetivo general de este proyecto es investigar si existen diferencias fenotípicas y genéticas entre los rizobios que nodulan Lupinus mariae-josephi y lupinos de suelos ácidos nativos de la Península Ibérica (Lupinus angustifolius, L.luteus y otras cuatro especies). En este proyecto se han aislado bacterias (rizobios) de nódulos de L. maria-josephi a partir de suelos básicos de Valencia (localidad de LLombai) y se está realizando su caracterización a nivel morfológico, nutricional, simbiótico y molecular. A nivel molecular se han comparado los genes “housekeeping” 16S rRNA, recA, atpD, gln2 y el simbiótico, nodC de diversas cepas que nodulan L. mariae-josephi y con cepas de otras especies de rizobios. Por otra parte, también se está evaluando la capacidad de L. mariae-josephi de ser nodulada por diferentes rizobios bajo condiciones bacteriológicamente controladas

Taxonomy of endosymbiotic bacteria from a novel Lupinus sp. (Lupinus mariae-josephi) endemic of a limed-alkaline soil habitat in Southeastern Spain

Lupinus mariae-josephi is a recently described Lupinus species (Pascual 2004) endemic of a Southeastern area of Spain with soils singularly of high pH and active lime content where it is endangered due to the reduced size of its habitat. Ten isolates of L. mariae-josephi endosymbiotic bacteria were obtained using trap-plants and soils from five sampling points within a native plant population area in Llombai (Valencia, Spain). The microsymbionts are extra-slow (ultrabradytrophic) growing bacteria with phenotypic and symbiotic characteristics singularly different from Bradyrhizobium strains nodulating other Lupinus spp. thriving in the Iberian Peninsula and adapted to growth in acidic soils. Cross-inoculation experiments revealed that these L. mariae-josephi endosymbiotic bacteria isolates are unable to nodulate or efficiently fix nitrogen with other Lupinus spp. Their phylogenetic status was examined by a multilocus sequence analysis of four housekeeping genes (16S rDNA, glnII, recA, atpD) and the symbiotic nodC gene. The 16S rDNA phylogenetic analysis showed that L. mariae-josephi isolates are related to strains nodulating Retama spp. in northeastern Algeria (Boulila et al., 2009), Phaseolus lunatus from Peru (Ormeño-Orrillo et al., 2006), as well as to B. elkanii, B. jicamae and B. pachyrhizi species, forming a new clade (Clade I) within the Bradyrhizobium genus. All the single and concatenated glnII+recA and glnII+recA+atpD analyses consistently support the existence of Clade I, and also revealed that, within this clade, the L. mariae-josephi endosymbiotic bacteria belong to a single evolutionary lineage that also includes strains nodulating Retama spp. from northeastern Algeria. Within this new Bradyrhizobium lineage, the phylogenetic analyses performed showed essentially convergent results indicating that the tested L. mariae-josephi isolates nested in three sub-groups that might correspond to novel sister Bradyrhizobium species. Bradyrhizobium Clade I is highly differentiated from the Bradyrhizobium clade (Clade II) that includes currently named Bradyrhizobium species and well-delineated unnamed genospecies. Singularly, all the endosymbiotic bacteria from Lupinus species adapted to acid soils in the Iberian Peninsula and tested in this study are included in Clade II. They are related either to strains of the B. canariense or B. japonicum lineages. The phylogenetic analysis based on the symbiotic nodC gene showed that L. mariae-josephi endosymbiotic bacteria define a novel branch in the nodC Bradyrhizobium tree. This branch groups together with a branch that gathers isolates from recently studied legume symbiosis such as isolates from Retama spp., which suggests the existence of a common unique ancestor for the symbiotic genes of these two groups of bradyrhizobia. In contrast, the symbiotic genes of isolates from other Lupinus spp. from the Iberian Peninsula are clearly related to the B. canariense lineage. The allopatric (geographic) speciation of the L. mariae-josephi bradyrhizobia may result from the colonization of a singular habitat, such as the basic and high calcium carbonate soils of the Valencia area, by its unique legume host

Phenotypic and phylogenetic characterization of endosymbiotic bacteria from Lupinus mariae-josephi

Lupinus mariae-josephi is a Lupinus species that thrives in a Southeastern area of Spain (Valencia) in soils of singularly high pH and active lime content. It is nodulated by extra-slow growing bacteria symbiotically and phylogenetically distant to endosymbiotic strains nodulating other Lupinus sp. native of the Iberian Peninsula and adapted to growth in acid soils. Cross-inoculation experiments revealed that the L. mariae-josephi endosymbiotic bacteria are unable to nodulate or efficiently fix nitrogen with well-known Lupinus spp. Their species affiliation was examined by a multilocus sequence analysis of four housekeeping genes (16S rDNA, glnII, recA, atpD) and the symbiotic nodC gene. Single and concatenated phylogenetic analyses of these genes consistently revealed that L. mariae-josephi endosymbiotic bacteria belong to a clade, within the Bradyrhizobium genus, highly differentiated from the Bradyrhizobium clade that includes currently named Bradyrhizobium species as well as the endosymbiotic bacteria from Lupinus species tested in this study. Within this new clade the L. mariae-josephi bacteria nested in several subgroup that may correspond to novel sister species. The phylogenetic analysis based on the nodC gene showed that L. mariae-josephi endosymbiotic bacteria define a novel branch of the nodC Bradyrhizobium tree and likely have a common unique ancestor for the symbiotic genes with nodule isolates from Retama spp

α-Hispanolol Induces Apoptosis and Suppresses Migration and Invasion of Glioblastoma Cells Likely via Downregulation of MMP-2/9 Expression and p38MAPK Attenuation

α-Hispanolol (α-H) is a labdane diterpenoid that has been shown to induce apoptosis in several human cancer cells. However, the effect of α-H in human glioblastoma cells has not been described. In the present work, we have investigated the effects of α-H on apoptosis, migration, and invasion of human glioblastoma cells with the aim of identifying the molecular targets underlying its mechanism of action. The results revealed that α-H showed significant cytotoxicity against human glioma cancer cell lines U87 and U373 in a concentration- and time-dependent manner. This effect was higher in U87 cells and linked to apoptosis, as revealed the increased percentage of sub-G1 population by cell cycle analysis and acquisition of typical features of apoptotic cell morphology. Apoptosis was also confirmed by significant presence of annexin V-positive cells and caspase activation. Pretreatment with caspase inhibitors diminishes the activities of caspase 8, 9, and 3 and maintains the percentage of viable glioblastoma cells, indicating that α-H induced cell apoptosis through both the extrinsic and the intrinsic pathways. Moreover, we also found that α-H downregulated the anti-apoptotic Bcl-2 and Bcl-xL proteins and activated the pro-apoptotic Bid and Bax proteins. On the other hand, α-H exhibited inhibitory effects on the migration and invasion of U87 cells in a concentration-dependent manner. Furthermore, additional experiments showed that α-H treatment reduced the enzymatic activities and protein levels of matrix metalloproteinase MMP-2 and MMP-9 and increased the expression of TIMP-1 inhibitor, probably via p38MAPK regulation. Finally, xenograft assays confirmed the anti-glioma efficacy of α-H. Taken together, these findings suggest that α-H may exert anti-tumoral effects in vitro and in vivo through the inhibition of cell proliferation and invasion as well as by the induction of apoptosis in human glioblastoma cells. This research describes α-H as a new drug that may improve the therapeutic efficacy against glioblastoma tumors.This study was supported by grant PI11/00036, PI14/00055, and PI17/00012 from the FIS, MPY 1410/09 from ISCIII and Spanish Ministry of Health (Instituto de Salud Carlos III; RD12/0036/0059) to SoH and by grants IERPY 1149/16 and IERPY-M 389/18 to AL. L JG was supported by FIS (FI12/00340). SaH was supported by IERPY 1149/16 from ISCIII.S

Emotional Processing Profile in Patients with First Episode Schizophrenia:The Influence of Neurocognition

This study sought to investigate the influence of neurocognition on the emotional processing profiles of patients with first-episode schizophrenia, using the 4-branch Mayer-Salovey-Caruso Emotional Intelligence Test (MSCEIT) (Perceiving Emotions; Facilitating Emotions; Understanding Emotions and Managing Emotions). A sample of 78 patients with first-episode schizophrenia and a group of 90 non-psychiatric control subjects were included in this work. The initial results showed that patients had lower scores than controls for the "Understanding Emotions" and "Managing Emotions" MSCEIT branches. However, after controlling for neurocognition, the only deficits were found on the "Managing Emotions" branch of the MSCEIT. This branch can be considered as measuring a more sophisticated level of emotional processing, which may constitute a deficit in itself. In conclusion, patients with first-episode schizophrenia present deficits in social cognition at the highest level that seem to be independent from neurocognition. These findings support the inclusion of the "Managing Emotions" branch of the MSCEIT as part of the MCCB.Instituto de Salud Carlos III (PI16/00359)Instituto de Salud Carlos III (PI19/00766)Comunidad de Madrid (S2017/BMD-3740)Ministry of Health (MOH-000013)4.242 JCR (2020) Q1, 39/167 Medicine, general & internal1.040 SJR (2021) Q1, 438/2489 Medicine (miscellaneous)No data IDR 2020UE

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

CsRAV1 induces sylleptic branching in hybrid poplar

• Sylleptic branching in trees may increase significantly branch number, leaf area and the general growth of the tree, particularly in its early years. Although this is a very important trait, so far little is known about the genes that control this process. • This article characterizes the Castanea sativa RAV1 gene, homologous to Arabidopsis TEM genes, by analyzing its circadian behavior and examining its winter expression in chestnut stems and buds. Transgenic hybrid poplars over-expressing CsRAV1 or showing RNA interference down-regulated PtaRAV1 and PtaRAV2 expression were produced and analyzed. • Over-expression of the CsRAV1 gene induces the early formation of sylleptic branches in hybrid poplar plantlets during the same growing season in which the lateral buds form. Only minor growth differences and no changes in wood anatomy are produced. • The possibility of generating trees with a greater biomass by manipulating the CsRAV1 gene makes CsRAV1 transgenic plants promising candidates for bioenergy production

Targeting Tumor Mitochondrial Metabolism Overcomes Resistance to Antiangiogenics

Epithelial malignancies are effectively treated by antiangiogenics; however, acquired resistance is a major problem in cancer therapeutics. Epithelial tumors commonly have mutations in the MAPK/Pi3K-AKT pathways, which leads to high-rate aerobic glycolysis. Here, we show how multikinase inhibitor antiangiogenics (TKIs) induce hypoxia correction in spontaneous breast and lung tumor models. When this happens, the tumors downregulate glycolysis and switch to long-term reliance on mitochondrial respiration. A transcriptomic, metabolomic, and phosphoproteomic study revealed that this metabolic switch is mediated by downregulation of HIF1α and AKT and upregulation of AMPK, allowing uptake and degradation of fatty acids and ketone bodies. The switch renders mitochondrial respiration necessary for tumor survival. Agents like phenformin or ME344 induce synergistic tumor control when combined with TKIs, leading to metabolic synthetic lethality. Our study uncovers mechanistic insights in the process of tumor resistance to TKIs and may have clinical applicability

Delayed introduction of sirolimus in paediatric intestinal transplant recipients: indications and long-term benefits

To review our experience using sirolimus in a single centre paediatric intestinal transplantation cohort. Intestinal transplant patients with more than 3 months follow-up were divided into two groups according to their immunosuppression regimen: tacrolimus, (TAC group, n = 45 grafts) or sirolimus (SRL group, n = 38 grafts), which included those partially or completely converted from tacrolimus to sirolimus. The indications to switch were tacrolimus side effects and immunological complications. Survival and complications were retrospectively analysed comparing both groups. SRL was introduced 9 months (0 months–16.9 years) after transplant. The main cause for conversion was worsening renal function (45%), followed by haemolytic anaemia (21%) and graft-versus-host-disease (16%). Both groups showed a similar overall patient/graft survival (P = 0.76/0.08) and occurrence of rejection (24%/17%, P = 0.36). Immunological complications did not recur after conversion. Renal function significantly improved in most SRL patients. After a median follow-up of 65.17 months, 28/46 survivors were on SRL, 26 with monotherapy, with good graft function. Over one-third of our patients eventually required SRL conversion that allowed to improve their kidney function and immunological events, without entailing additional complications or survival impairment. Further trials are warranted to clarify the potential improvement of the standard tacrolimus maintenance by sirolimus conversion or addition.Fil: Andres, Ane M.. Hospital Universitario la Paz; EspañaFil: Talayero, Paloma. Hospital Universitario 12 de Octubre; EspañaFil: Alcolea Sanchez, Alida. No especifíca;Fil: Sanchez Galán, Alba. Hospital Universitario la Paz; EspañaFil: Serradilla Rodríguez, Javier. Hospital Universitario la Paz; EspañaFil: Bueno Jimenez, Alba. Hospital Universitario la Paz; EspañaFil: Gonzalez Sacristan, Rocío. No especifíca;Fil: Stringa, Pablo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; ArgentinaFil: Papa Gobbi, Rodrigo. Hospital Universitario la Paz; EspañaFil: Lasa Lazaro, Maria. Hospital Universitario 12 de Octubre; EspañaFil: Díaz Almirón, Mariana. Hospital Universitario la Paz; EspañaFil: Ramos Boluda, Esther. No especifíca;Fil: Lopez Santamaría, Manuel. Hospital Universitario la Paz; EspañaFil: Hernández Oliveros, Francisco. Hospital Universitario la Paz; Españ