Plant growth promotion of the forage plant Lupinus albus Var. Orden Dorado using Pseudomonas agronomica sp. nov. and Bacillus pretiosus sp. nov. added over a valorized agricultural biowaste

IntroductionThe overexploitation of natural ecosystems and the evolution of climate change currently force us to design new strategies for more sustainable agronomic uses. The recovery of plant residues, as an alternative to agrochemicals, can help alleviate these problems, for example, through its use for the synthesis of biofertilizers. In this work, the effect of the organic fertilizer matrix ORGAON® from the valorization of horticultural waste is tested, to which two strains of bacteria (and their consortium) are added (SAICEU11T identified as Bacillus pretiosus and SAICEU22T identified as Pseudomonas agronomica), selected for their demonstrated ability to promote plant growth (PGPB), on the lupine forage plant (Lupinus albus).MethodsFor the synthesis of the biofertilizer, both strains were added to the ORGAON® organic matrix separately, until reaching a final optical density (OD) of 0.5 McFarland in each case in the irrigation matrix. As a control, sterile ORGAON® (ORGAON®st) was used, also supplemented with the PGPB strains and a chemical fertilizer widely used in agronomy (Chem-F). With these treatments, a 6-week experiment was started under controlled laboratory conditions and on agricultural substrate, to recreate field conditions as accurately as possible. All the tests were carried out with 9 repetitions and 3 replicates of each treatment. After harvest, the improvements on the following biometric variables were studied for each treatment: total weight (Weight_T, g), shoot weight (Weight_S, g), root weight (Weight_R, g), number of leaves (Leaves, No.), shoot length (Length_S), root length (Length_R) and number of secondary roots (Roots, No.). Likewise, the identification of the tested strains and their description as new species was carried out. For this, they were studied from the phenotypic point of view (Transmission electron microscopy (TEM), metabolic profile, PGP activities, fatty acid profile and Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF)) and genotypic (sequencing of the main housekeeping genes and sequencing of the whole genome, genomic characteristics (dDDH and ANI) and phylogenetic analysis).Results and discussionAfter the statistical analysis of the results, it is shown that the individual addition of both strains on the ORGAON® and ORGAON®st organic matrix improve certain biometric variables. In the case of the SAICEU11T (Bacillus pretiosus) strain, the variables root weight (Weight_R, g), total weight (Weight_T, g) and length of the plant, and number of secondary roots (Roots, No.) significantly improve, while in the case of the strain SAICEU22T (Pseudmonas agronomica), a significant improvement of root length (Length_R) and number of secondary roots (Roots, No.) is demonstrated. On the other hand, the genotaxonomic analysis showed that both species have not been described to date. The identification based on the main housekeeping genes, show that for the Bacillus strain (SAICEU11T) the sequence similarity of the 16S rRNA was 100%, gyrB 92.69%, rpoB 97.70% and rpoD 94.67%. For the Pseudomonas strain (SAICEU22T) the results were 100% for 16S rRNA, 98.43% for rpoD and 96.94% for gyrB. However, in both cases, the dDDH and ANI values, as well as the phylogenetic analysis, show that both species are below the species threshold, which would support the hypothesis that both are new species, in line with the chemotaxonomic results obtained by MALDI-TOF spectrometry and fatty acid profile. To verify the biosafety in their handling and release into the natural environment, we have ruled out the presence of genes that encode virulence factors or resistance to antibiotics, concluding that they are suitable for use in the field to improve the yield of crop plants. Type strains are SAICEU11T (= DSM 114702T = CECT30674T) for Bacillus pretiosus and SAICEU22T (= DSM 114959T = CECT30673T) for Pseudomonas agronomicae

Risk factors for premature aging of placenta: comparative study of perinatal outcomes between grannum grade III placentas and grannum grade I-II placentas

Introduction: The placenta aging has been related with intrauterine fetal growth, low maternal age, Caucasian, multiparity, hypertensive states and smoking habit. Grannum P. classification is the most used for its assessment. The association between grade III placenta (G3P) and ex-smoking or smokeexposed pregnants has not been studied Main outcome: To asses if smoking, being an ex-smoker or a passive-smoker is a risk factor for developing grade III placenta, as well as if there is a smoking-free period of time to avoid the effect of smoking over placenta Material and methods: A retrospective case-control study of single pregnancies followed-up at the Obstetric Ultrasound Unit between January 2013 and January 2014. Placental grading according to Grannum classification was stablished through abdominal approach between 34-36 weeks of gestation and two groups were established: grade III placenta and grade I-II placenta (G1-2P) Maternal and paternal characteristics, type of delivery and perinatal outcomes were collected Results: Baseline characteristics were similar between the two groups. In G3P the incidence of hypertensive disease of pregnancy was higher(p=0,0107). The percentage was similar for premature birth, 1st and 5th minute Apgar, type of delivery and cesarean due to risk of loss of fetal wellbeing. A lower neonatal weight was found in G3P, at the same median days at delivery, with a mean difference of 148,156(p=0,008313. Regarding weight percentile, it was found a p35 in G3P and a p47,5 in G1- 2P(p=0,08235) 15% of the total pregnant were smokers. In G3P group it was found a higher frequency of smokers and ex-smokers since 1st trimester of pregnancy (p=0.0001), as well as pregnant non-smokers with an smoking partner(p=0,0001). There was an strong evidence for association between pregnant smokers and smoking partners(p=0,0001). No difference was found regarding to neonatal weight comparing smokers, ex-smokers and ex-smokers since 1st trimester of pregnancy. The length of pregnancy was lower between G3P pregnant smokers compared to G1-2P pregnant smokers, with a mean difference of 8 days (p=0,00091) Conclusions: There is a strong evidence for association between smoking during pregnancy, quitting smoking at the beginning of the pregnancy or being a passive smoker with development of G3P Some pregnant smokers don`t develop premature aging of placenta, it could be due to either other parameters or a later aging (data were collected between 34-36 weeks). The association between quitting smoking at the beginning of the pregnancy and not having a smoking partner reduces the risk of developing G3

Oxidative stress protection and growth promotion activity of Pseudomonas mercuritolerans sp. nov., in forage plants under mercury abiotic stress conditions

SAICEUPSMT strain was isolated from soils in the mining district of Almadén (Ciudad Real, Spain), subjected to a high concentration of mercury. Using the plant model of lupinus, the strain was inoculated into the rhizosphere of the plant in a soil characterized by a high concentration of mercury (1,710 ppm) from an abandoned dump in the mining district of Almadén (Ciudad Real, Spain). As a control, a soil with a minimum natural concentration of mercury, from a surrounding area, was used. Under greenhouse conditions, the effect that the inoculum of the SAICEUPSMT strain had on the antioxidant capacity of the plant was studied, through the quantification of the enzymatic activity catalase (CAT), ascorbate peroxidase (APX), superoxide dismutase (SOD), and glutathione reductase (GR). Likewise, the capacity of the plant to bioaccumulate mercury in the presence of the inoculum was studied, as well as the effect on the biometric parameters total weight (g), shoot weight (g), root weight (g), shoot length (cm), root length (cm), total number of leaves (N), and total number of secondary roots (No). Finally, in view of the results, the SAICEUPSMT strain was identified from the phenotypic and genotypic point of view (housekeeping genes and complete genome sequencing). The inoculum with the SAICEUPSMT strain in the presence of mercury produced a significant reduction in the enzymatic response to oxidative stress (CAT, APX, and SOD). It can be considered that the strain exerts a phytoprotective effect on the plant. This led to a significant increase in the biometric parameters total plant weight, root weight and the number of leaves under mercury stress, compared to the control without abiotic stress. When analyzing the mercury content of the plant with and without bacterial inoculum, it was found that the incorporation of the SAICEUPSMT strain significantly reduced the uptake of mercury by the plant, while favoring its development in terms of biomass. Given the positive impact of the SAICEUPSMT strain on the integral development of the plant, it was identified, proving to be a Gram negative bacillus, in vitro producer of siderophores, auxins and molecules that inhibit stress precursors. The most represented fatty acids were C16:0 (33.29%), characteristic aggregate 3 (22.80%) comprising C16:1 ω7c and C16: 1ω6c, characteristic aggregate 8 (13.66%) comprising C18:1 ω7c, and C18: 1 cycle ω6c and C 17:0 (11.42%). From the genotypic point of view, the initial identification of the strain based on the 16S rRNA gene sequence classified it as Pseudomonas iranensis. However, genome-wide analysis showed that average nucleotide identity (ANI, 95.47%), DNA-DNA in silico hybridization (dDDH, 61.9%), average amino acid identity (AAI, 97.13%), TETRA (0.99%) and intergenic distance (0.04) values were below the established thresholds for differentiation. The results of the genomic analysis together with the differences in the phenotypic characteristics and the phylogenetic and chemotaxonomic analysis support the proposal of the SAICEUPSMT strain as the type strain of a new species for which the name Pseudomonas mercuritolerans sp. is proposed. No virulence genes or transmissible resistance mechanisms have been identified, which reveals its safety for agronomic uses, under mercury stress conditions

Análisis de la situación de los estudios agronómicos y forestales en Europa. Anexo III: análisis de los estudios europeos

El documento que se presenta es el resultado del análisis de la situación actual en estas ingenierías y del estudio del futuro de las mismas. Todas las propuestas que aquí se hacen se han consensuado en las sesiones del Plenario del grupo de trabajo. Por ello, es justo reconocer la aportación que este trabajo puede suponer a la futura Convergencia Europea en la Educación Superior

Sprouty2 and Spred1-2 Proteins Inhibit the Activation of the ERK Pathway Elicited by Cyclopentenone Prostanoids

Sprouty and Spred proteins have been widely implicated in the negative regulation of the fibroblast growth factor receptor-extracellular regulated kinase (ERK) pathway. In considering the functional role of these proteins, we explored their effects on ERK activation induced by cyclopentenone prostanoids, which bind to and activate Ras proteins. We therefore found that ectopic overexpression in HeLa cells of human Sprouty2, or human Spred1 or 2, inhibits ERK1/2 and Elk-1 activation triggered by the cyclopentenone prostanoids PGA1 and 15d-PGJ2. Furthermore, we found that in HT cells that do not express Sprouty2 due to hypermethylation of its gene-promoter, PGA1-provoked ERK activation was more intense and sustained compared to other hematopoietic cell lines with unaltered Sprouty2 expression. Cyclopentenone prostanoids did not induce Sprouty2 tyrosine phosphorylation, in agreement with its incapability to activate tyrosine-kinase receptors. However, Sprouty2 Y55F, which acts as a defective mutant upon tyrosine-kinase receptor stimulation, did not inhibit cyclopentenone prostanoids-elicited ERK pathway activation. In addition, Sprouty2 did not affect the Ras-GTP levels promoted by cyclopentenone prostanoids. These results unveil both common and differential features in the activation of Ras-dependent pathways by cyclopentenone prostanoids and growth factors. Moreover, they provide the first evidence that Sprouty and Spred proteins are negative regulators of the ERK/Elk-1 pathway activation induced not only by growth-factors, but also by reactive lipidic mediators

Thin film nanostructuring at oblique angles by substrate patterning

It is demonstrated that, besides classical nanocolumnar arrays, the oblique angle geometry induces the growth of singular structures in the nanoscale when using wisely designed patterned substrates. Well-ordered array of crosses, cylindrical nanorods or hole structures arranged in square or hexagonal regular geometries are reported as examples, among others. The fundamental framework connecting substrate topography and film growth at oblique angles is presented, allowing the use of substrate patterning as a feasible thin film nanostructuring technique. A systematic analysis of the growth of TiO2 thin films on 4 different lithographic patterned substrates in 4 different scale lengths is also presented. A first conclusion is the existence of a height-based selective growth in the initial stages of the deposition, by which the film preferentially develops on top of the tallest substrate features. This behavior is maintained until the film reaches a critical thickness, the so-called Oblivion Thickness, above which the film topography becomes gradually independent of the substrate features. A general formula relating the spatial features of the pattern, the coarsening exponent and the Oblivion Thickness has been deduced.The authors thank the financial support from MCIN/AEI/10.13039/ 501100011033 projects PID2019-110430GB-C21, PID2020-112620GBI00, PID2020-114270RA-I00 and RTI2018-098117-B-C21 (also financed by FEDER Una manera de hacer europa), the Junta de Andalucía (PAIDI- 2020 through projects P18-RT-3480 and P18-RT-6079, and through its 2019 PhD Researcher Hiring Program), the CSIC (2019AEP161 and 201860E050), the Regional Government of Madrid (project IND2017/ IND-7668 and YEI contract PEJ-2019-AI/IND-14451 with support from FSE), the H2020-EU.1.2.1-FET OPEN program (grant 899352, project SOUNDofICE, and the EFRE Infra-Pro project ChAMP), and the University of Seville (VI PPIT-US). The work is supported by the Deutsche Forschungsgemeinschaft (DFG, grant Scha 632/24, “Tailored Disorder” and Scha 632/27, “DFGGer ¨atezentrum”). This work is also supported by the free state of Thuringia under grants 2015 FGI 0025 305 (FastμXRD) and B715-10009 (BioMacroNano2020), all co-financed by the European Union within the framework of the European Regional Development Fund (ERDF). The service from the MiNa Laboratory at IMN-CNM (CSIC), funded from CM (project S2018/NMT-4291 TEC2SPACE), MINECO (project CSIC13-4E- 1794) and EU (FEDER, FSE), is also acknowledged.Peer reviewe

Activation of a Ca2+-permeable cation channel by two different inducers of apoptosis in a human prostatic cancer cell line

We have combined patch clamp recording with simultaneous [Ca2+]i measurements in single LNCaP cells (a human prostate cancer cell line), to study the activation of Ca2+-permeable channels by two different inducers of apoptosis, ionomycin and serum deprivation.In perforated patch recording, LNCaP cells had a membrane potential of -40 mV and a resting [Ca2+]i of 90 nM. Application of ionomycin at levels that induced apoptosis in these cells (10 μM) produced a biphasic increase in [Ca2+]i. The first rise in [Ca2+]i was due to release of Ca2+ from internal stores and it was associated with a membrane hyperpolarization to -77 mV. The latter was probably due to the activation of high conductance, Ca2+- and voltage-dependent K+ channels (maxi-K). Conversely, the second rise in [Ca2+]i was always preceded by and strictly associated with membrane depolarization and required external Ca2+. Serum deprivation, another inducer of apoptosis, unmasked a voltage-independent Ca2+ permeability as well.A lower concentration of ionomycin (1 μM) did not induce apoptosis, and neither depolarized LNCaP cells nor produced the biphasic increase in [Ca2+]i. However, the first increment in [Ca2+]i due to release from internal Ca2+ stores was evident at this concentration of ionomycin.Simultaneous recordings of [Ca2+]i and ion channel activity in the cell attached configuration of patch clamp revealed a Ca2+-permeable, Ca2+-independent, non-selective cation channel of 23 pS conductance. This channel was activated only during the second increment in [Ca2+]i induced by ionomycin. The absence of serum activated the 23 pS channel as well, albeit at a lower frequency than with ionomycin.Thus, the 23 pS channel can be activated by two unrelated inducers of apoptosis and it could be another Ca2+ influx mechanism in programmed cell death of LNCaP cells

Verso heroico en loor del verdadero heroe en campaña Dn. Pedro Agustin de Echevarría, comandante del batallon franco y sus individuos desde la formacion de este cuerpo

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The Neurobiology of Behavior and Its Applicability for Animal Welfare: A Review

Understanding the foundations of the neurobiology of behavior and well-being can help us better achieve animal welfare. Behavior is the expression of several physiological, endocrine, motor and emotional responses that are coordinated by the central nervous system from the processing of internal and external stimuli. In mammals, seven basic emotional systems have been described that when activated by the right stimuli evoke positive or negative innate responses that evolved to facilitate biological fitness. This review describes the process of how those neurobiological systems can directly influence animal welfare. We also describe examples of the interaction between primary (innate) and secondary (learned) processes that influence behavior