Redox activities and ROS, NO and phenylpropanoids production by axenically cultured intact olive seedling roots after interaction with a mycorrhizal or a pathogenic fungus

Las raíces de las plántulas de olivo, en cultivo axénico, fueron colocadas alternativamente en contacto con Rhizophagus irregulares (micorrícicos) o con hongos Verticillim dahliae (patógenos). También se incluyeron tratamientos MeJA. Las raíces intactas (generación de anión superóxido, superóxido dismutasa y actividades de peroxidasa) se midieron en las actividades in vivo del apoplasto. Todos nuestros resultados mostraron que las actividades redox apoplásticas de raíces de las plántulas intactas en contacto con el hongo micorriza compatible fueron claramente atenuados en comparación con el hongo patógeno o tratado con MeJA, incluso en las primeras etapas usadas en el tratamiento. Los fenoles totales, flavonoides y glucósidos fenilpropanoides, también fueron cuantificados. Las raíces en contacto con el hongo micorriza no mejoraron la biosíntesis de compuestos fenólicos con respecto a los controles, mientras que los de contacto con el patógeno mejoraron de forma significativa la biosíntesis de todas las fracciones fenólicas medidas. Las especies reactivas del oxígeno y la acumulación de óxido nítrico en las raíces fueron examinadas por microscopía de fluorescencia. Todos ellas presentaron una acumulación mucho mayor en las raíces en contacto con el patógeno que con el hongo micorriza. En total, estos resultados indican que las raíces de las plántulas intactas de olivo, claramente diferenciadas entre micorrizas y hongos patógenos, atenuan las reacciones de defensa contra la primera para facilitar su creación, mientras que induce una reacción de defensa fuerte y sostenida contra el segundo. Ambas especies reactivas de oxígeno y nitrógeno parecían estar involucrados en estas respuestas desde los primeros momentos de contacto. Sin embargo, se necesitan más investigaciones para aclarar la diafonía propuesta entre ellos y sus respectivas funciones en estas respuestas ya que las imágenes de fluorescencia de las raíces revelaron que las especies reactivas del oxígeno se acumulan principalmente en el apoplasto (congruente con las actividades redox medidas en este compartimento), mientras el óxido nítrico se almacena principalmente en el citosol.Roots of intact olive seedlings, axenically cultured, were alternatively placed in contact with Rhizophagus irregularis (mycorrhizal) or Verticillim dahliae (pathogenic) fungi. MeJA treatments were also included. In vivo redox activities in the apoplast of the intact roots (anion superoxide generation, superoxide dismutase and peroxidase activities) were measured. All our results showed that apoplastic redox activities of intact seedling roots in contact with the compatible mycorrhizal fungus were clearly attenuated in comparison with the pathogenic fungus or treated with MeJA, even at the early stages of treatment used. Total phenolics, flavonoids and phenylpropanoid glycosides were also quantified. Roots in contact with the mycorrhizal fungus did not enhance the biosynthesis of phenolic compounds with respect to controls, while those in contact with the pathogenic one significantly enhanced the biosynthesis of all phenolic fractions measured. Reactive oxygen species and nitric oxid accumulation in roots were examined by fluorescence microscopy. All of them presented much higher accumulation in roots in contact with the pathogenic than with the mycorrhizal fungus. Altogether these results indicate that intact olive seedling roots clearly differentiated between mycorrhizal and pathogenic fungi, attenuating defense reactions against the first to facilitate its establishment, while inducing a strong and sustained defense reaction against the second. Both reactive oxygen and nitrogen species seemed to be involved in these responses from the first moments of contact. However, further investigations are required to clarify the proposed crosstalk between them and their respective roles in these responses since fluorescence images of roots revealed that reactive oxygen species were mainly accumulated in the apoplast (congruently with the measured redox activities in this compartment) while nitric oxid was mainly stored in the cytosol.-- Ministerio de Ciencia e Innovación. Proyecto CGL2009-12406 -- Junta de Extremadura. Proyecto PRI09A023peerReviewe

A window to the sea: environmental indicators for coastal risk management under the RAIA observatory (NW-Iberian Peninsula)

ECSA 59 Using the best scientific knowledge for the sustainable management of estuaries and coastal seas, San Sebastian (Spain), 5th–8th September 2022The international RAIA Observatory (www.marnaraia.org) resulted from the effort of 12 research and academic institutions and public agencies (Spanish and Portuguese) working in the field of meteorology and oceanography. The RAIA Observatory serves the main maritime activities of the Galicia-Northern Portugal Euroregion and contributes to collaborative observational networks. Under the framework of coastal risk management, environmental indicators are fundamental tools for the evaluation and mitigation of environmental risks, showing the current state and helping to predict future changes on ecosystem health regarding environmental risks. In last years, the different partners of the RAIA Observatory have identified 38 environmental indicators, in which 12 key risks affecting the ecosystem services of the Galicia-Northern Portugal Euroregion are being evaluated. Data was analyzed and compiled by the various partners of the RAIA Observatory and the development and optimization of the environmental indicators has been done according to the specifications provided by the European Environmental Agency (EEA) and the International Panel for Climate Change (IPCC). The resulting environmental indicators are included and shared on a publicly-accessible Web service, georeferenced and accompanied by plots (https://marrisk.inesctec.pt/public/#!/indicators). So far, the current status of the indicators has allowed us to establish risk assessment protocols for the Euroregion, and identify critical gaps in a temporal and spatial coverage. The inclusion of the environmental indicators in the RAIA Observatory is of great relevance for national and international data exchange and promotes future collaborationsN

Hippocampal-Dependent Spatial Memory in the Water Maze is Preserved in an Experimental Model of Temporal Lobe Epilepsy in Rats

Cognitive impairment is a major concern in temporal lobe epilepsy (TLE). While different experimental models have been used to characterize TLE-related cognitive deficits, little is known on whether a particular deficit is more associated with the underlying brain injuries than with the epileptic condition per se. Here, we look at the relationship between the pattern of brain damage and spatial memory deficits in two chronic models of TLE (lithium-pilocarpine, LIP and kainic acid, KA) from two different rat strains (Wistar and Sprague-Dawley) using the Morris water maze and the elevated plus maze in combination with MRI imaging and post-morten neuronal immunostaining. We found fundamental differences between LIP- and KA-treated epileptic rats regarding spatial memory deficits and anxiety. LIP-treated animals from both strains showed significant impairment in the acquisition and retention of spatial memory, and were unable to learn a cued version of the task. In contrast, KA-treated rats were differently affected. Sprague-Dawley KA-treated rats learned less efficiently than Wistar KA-treated animals, which performed similar to control rats in the acquisition and in a probe trial testing for spatial memory. Different anxiety levels and the extension of brain lesions affecting the hippocampus and the amydgala concur with spatial memory deficits observed in epileptic rats. Hence, our results suggest that hippocampal-dependent spatial memory is not necessarily affected in TLE and that comorbidity between spatial deficits and anxiety is more related with the underlying brain lesions than with the epileptic condition per se

A window to the sea: environmental indicators for coastal risk management under the RAIA observatory (NW-Iberian Peninsula)

Poster.-- International Ocean Data Conference 2022, Sopot, Poland, 14-16 February 2022Environmental indicators are fundamental tools for the evaluation and mitigation of environmental risks, showing the current state and helping to predict future changes on ecosystem health regarding environmental risks The framework of the RAIA observatory www marnaraia org identified 38 environmental indicators, in which 12 key risks affecting the ecosystem services of the Euroregion Galicia Northern Portugal have been evaluated The objective of this initiative is to optimize and analyze the environmental indicators identified for the Euroregion and make them available through a Web service that allows users to get information on the current state and evolution of the ecosystem healthThis contribution has been funded by the European Union MarRISK project Adaptación costera ante el Cambio Climático conocer los riesgos y aumentar la resiliencia (0262_MarRISK1_E) through EP-INTERREG V A España-Portugal (POCTEP) programN

Russian hegemonies: historical snapshots, regional security and changing forms of Russia's role in the post-Soviet region

The broad purpose of this study is to exemplify changing forms of hierarchical rule manifested in post-Soviet Russia's varying provision of regional security related to military conflicts. Russia's regional role varies in form and in thickness. Although the endurance of Soviet legacy can be observed in four major areas (stabilising borders, economic interest, unification of fellow Slavs and fluctuating alliances), there is enough variance to suggest that over-deterministic theories about Russian regional foreign policy do not account for conjunctural factors that can contribute to change. Instead, we can view the Soviet and post-Soviet regional dynamic as one that is on a fluid anarchy–hierarchy spectrum

In vitro and in vivo effects of an anti-mouse endoglin (CD105)-immunotoxin on the early stages of mouse B16MEL4A5 melanoma tumours

11 p.-6 fig.TGF-beta superfamily co-receptors are emerging as targets for cancer therapy, acting both directly on cells and indirectly on the tumour neovasculature. Endoglin (CD105), an accessory component of the TGF-beta receptor complex, is expressed in certain melanoma cell lines and the endothelial cells of tumour neovessels. Targeting endoglin with immunotoxins is an attractive approach for actively suppressing the blood supply to tumours. Here, we report evidence indicating that endoglin is expressed in mouse melanoma B16MEL4A5 and mouse fibroblast L929 cell lines. We prepared an immunotoxin to target endoglin by coupling the rat anti-mouse MJ7/18 (IgG2a) monoclonal antibody (mAb) to the non-toxic type 2 ribosome-inactivating protein nigrin b (Ngb) with N-succinimidyl 3-(2-pyridyldithio)-propionate (SPDP) as a linker with a molar nigrin b at a MJ7/18 stoichiometry of 2:1. The MJ7-Ngb immunotoxin generated killed both cell lines, with IC50 values of 4.2 × 10−9 M for B16MEL4A5 and 7.7 × 10−11 M for L929 cells. For in vivo assays of the immunotoxin, B16MEL4A5 cells were injected subcutaneously into the right flanks of 6-week-old C57BL/6 J mice. When the animals developed palpable solid tumours, they were subjected to treatment with the immunotoxin. While treatment with either MJ7/18 mAb or Ngb did not affect tumour development, treatment with the immunotoxin completely and steadily blocked tumour growth up to 7 days, after which some tumours re-grew. Thus, vascular-targeting therapy with this anti-vascular immunotoxin could promote the destruction of newly created tumour vessels at early stages of B16MEL4A5 tumour development and readily accessible CD105+ B16MEL4A5 melanoma cells.This research was supported by grants from the Junta de Castilla y León (Grupo de Excelencia GR106 and Convenio-Consejería de Sanidad) and UVA-GIR funding to T.G., Fondo de Investigaciones Sanitarias FISPI02/0917 to R.M. and FISPI04/1279 to J.M.F., Ministerio de Ciencia e Innovación of Spain (SAF2010-19222 to C.B.) and Genoma España (MEICA) to C.B.Peer reviewe

Molecular Mechanisms Involved in the Response to Desiccation Stress and Persistence in Acinetobacter baumannii

Desiccation tolerance contributes to the maintenance of bacterial populations in hospital settings and may partly explain its propensity to cause outbreaks. Identification and relative quantitation of proteins involved in bacterial desiccation tolerance was made using label-free quantitation and iTRAQ labeling. Under desiccating conditions, the population of the Acinetobacter baumannii clinical strain AbH12O-A2 decreased in the first week, and thereafter, a stable population of 0.5% of the original population was maintained. Using label-free quantitation and iTRAQ labeling, 727 and 765 proteins, respectively, were detected; 584 of them by both methods. Proteins overexpressed under desiccation included membrane and periplasmic proteins. Proteins associated with antimicrobial resistance, efflux pumps, and quorum quenching were overexpressed in the samples subjected to desiccation stress. Electron microscopy revealed clear morphological differences between desiccated and control bacteria. We conclude that A. baumannii is able to survive long periods of desiccation through the presence of cells in a dormant state, via mechanisms affecting control of cell cycling, DNA coiling, transcriptional and translational regulation, protein stabilization, antimicrobial resistance, and toxin synthesis, and that a few surviving cells embedded in a biofilm matrix are able to resume growth and restore the original population in appropriate environmental conditions following a “bust-and-boom” strategy

Haplo-Cord Transplantation Using CD34+ Cells from a Third-Party Donor to Speed Engraftment in High-Risk Patients with Hematologic Disorders

AbstractAmong the strategies to optimize engraftment of cord blood (CB) stem cell transplantation (SCT), single CB with the coinfusion of CD34+ stem cells from an HLA-mismatched auxiliary donor (haplo-cord) provides a valid alternative for adult patients without a suitable donor. A total of 132 high-risk adult patients with hematological malignancies from 3 Spanish institutions underwent myeloablative haplo-cord SCT. The median age was 37 years and median weight was 70 kg; 37% had active disease. The median number of postprocessing CB total nucleated and CD34+ cells was 2.4 × 107/kg (interquartile range [IQR], 1.8 to 2.9) and 1.4 × 105/kg (IQR, .9 to 2), respectively. Neutrophil engraftment occurred in a median of 11.5 days (IQR, 10.5 to 16.5) and platelet engraftment at 36 days (IQR, 25.5 to 77). Graft failure was 2% overall and only 9% for CB. Cumulative incidence of acute graft-versus-host disease (GHVD) grades II to IV was 21% and cumulative incidence of chronic GVHD was 21%. Median follow-up was 60 months (range, 3.5 to 163). Overall survival was 43.5%, event-free survival was 38.3%, nonrelapse mortality was 35%, and relapse was 20% at 5 years. Myeloablative haplo-cord SCT results in fast engraftment of neutrophils and platelets, low incidences of acute and chronic GVHD, and favorable long-term outcomes using single CB units with relatively low cell content. Moreover, CB cell dose had no impact on CB engraftment and survival in this study. Therefore, haplo-cord SCT expands donor availability while reducing CB cell dose requirements

Biofilm formation assays.

<p>A) Quantification of biofilm formation by <i>A</i>. <i>baumannii</i> ATCC 17978 (17978), <i>A</i>. <i>baumannii</i> ATCC 17978 Δ13573 (Δ13573), <i>A</i>. <i>baumannii</i> ATCC 17978 harbouring pETRA (17978 with empty pETRA), <i>A</i>. <i>baumannii</i> ATCC 17978 harbouring pETRA with sRNA 13575 (13573), and <i>A</i>. <i>baumannii</i> ATCC Δ13573 harbouring pETRA with sRNA 13575 (Δ13573 complemented). B) SEM analysis of <i>A</i>. <i>baumannii</i> ATCC 17978 (a), <i>A</i>. <i>baumannii</i> sRNA 13573 over-producing strain (b) and <i>A</i>. <i>baumannii</i> ATCC 17978 Δ13573 (c). Micrographs were taken at 5,000x and bars indicate the scale marks (2 μm).</p

Expression level of the sRNA 13573 coding region.

<p>Expression level of the sRNA 13573 coding region.</p