A genetic toolkit and gene switches to limit Mycoplasma growth for biosafety applications

Mycoplasmas have exceptionally streamlined genomes and are strongly adapted to their many hosts, which provide them with essential nutrients. Owing to their relative genomic simplicity, Mycoplasmas have been used to develop chassis for biotechnological applications. However, the dearth of robust and precise toolkits for genomic manipulation and tight regulation has hindered any substantial advance. Herein we describe the construction of a robust genetic toolkit for M. pneumoniae, and its successful deployment to engineer synthetic gene switches that control and limit Mycoplasma growth, for biosafety containment applications. We found these synthetic gene circuits to be stable and robust in the long-term, in the context of a minimal cell. With this work, we lay a foundation to develop viable and robust biosafety systems to exploit a synthetic Mycoplasma chassis for live attenuated vectors for therapeutic applications

The role of the cerebellum in drug-cue associative memory: functional interactions with the medial prefrontal cortex

Drug-induced Pavlovian memories are thought to be crucial for drug addiction because they guide behaviour towards environments with drug availability. Drug-related memory depends on persistent changes in dopamine-glutamate interactions in the medial pre- frontal cortex (mPFC), basolateral amygdala, nucleus accumbens core and hippocampus. Recent evidence from our laboratory indi- cated that the cerebellum is also a relevant node for drug-cue associations. In the present study, we tested the role that specific regions of the cerebellum and mPFC play in the acquisition of cocaine-induced preference conditioning. Quinolinic acid was used to manage a permanent deactivation of lobule VIII in the vermis prior to conditioning. Additionally, lidocaine was infused into the prelim- bic and infralimbic (IL) cortices for reversible deactivation before every training session. The present findings show, for the first time, that the cerebellum and mPFC might act together in order to acquire drug-cue Pavlovian associations. Either a dorsal lesion in lobule VIII or an IL deactivation encouraged cocaine-induced preference conditioning. Moreover, simultaneous IL-cerebellar deactivation prevented the effect of either of the separate deactivations. Therefore, similar to the IL cortex, neural activity in the cerebellum may be crucial for ensuring inhibitory control of the expression of cocaine-related memories

Mapping cFos expression after cerebellar and medial prefrontal deactivations in rats trained to acquire cocaine-induced preference conditioning

6th Mediterranean Conference of Neuroscience (MNS Malta 2017

From back to front: A functional model for the cerebellar modulation in the establishment of conditioned preferences for cocaine‐related cues

This is the pre-peer reviewed version of the following article: From back to front: A functional model for the cerebellar modulation in the establishment of conditioned preferences for cocaine‐related cues, which has been published in final form at https://doi.org/10.1111/adb.12834. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.It is now increasingly clear that the cerebellum may modulate brain functions altered in drug addiction. We previously demonstrated that cocaine‐induced conditioned preference increased activity at the dorsal posterior cerebellar vermis. Unexpectedly, a neurotoxic lesion at this region increased the probability of cocaine‐induced conditioned preference acquisition. The present research aimed at providing an explanatory model for such as facilitative effect of the cerebellar lesion. First, we addressed a tracing study in which we found a direct projection from the lateral (dentate) nucleus to the ventral tegmental area (VTA) that also receives Purkinje axons from lobule VIII in the vermis. This pathway might control the activity and plasticity of the cortico‐striatal circuitry. Then we evaluated cFos expression in different regions of the medial prefrontal cortex and striatum after a lesion in lobule VIII before conditioning. Additionally, perineuronal net (PNN) expression was assessed to explore whether the cerebellar lesion might affect synaptic stabilization mechanisms in the medial prefrontal cortex (mPFC). Damage in this region of the vermis induced general disinhibition of the mPFC and striatal subdivisions that receive dopaminergic projections, mainly from the VTA. Moreover, cerebellar impairment induced an upregulation of PNN expression in the mPFC. The major finding of this research was to provide an explanatory model for the function of the posterior cerebellar vermis on drug‐related memory. In this model, damage of the posterior vermis would release striatum‐cortical networks from the inhibitory tonic control exerted by the cerebellar cortex over VTA, thereby promoting drug effects

Nucleus incertus projections to rat medial septum and entorhinal cortex: rare collateralization and septal-gating of temporal lobe theta rhythm activity

Nucleus incertus (NI) neurons in the pontine tegmentum give rise to ascending forebrain projections and express the neuropeptide relaxin-3 (RLN3) which acts via the relaxin-family peptide 3 receptor (RXFP3). Activity in the hippocampus and entorhinal cortex can be driven from the medial septum (MS), and the NI projects to all these centers, where a prominent pattern of activity is theta rhythm, which is related to spatial memory processing. Therefore, we examined the degree of collateralization of NI projections to the MS and the medial temporal lobe (MTL), comprising medial and lateral entorhinal cortex (MEnt, LEnt) and dentate gyrus (DG), and the ability of the MS to drive entorhinal theta in the adult rat. We injected fluorogold and cholera toxin-B into the MS septum and either MEnt, LEnt or DG, to determine the percentage of retrogradely labeled neurons in the NI projecting to both or single targets, and the relative proportion of these neurons that were RLN3-positive ( +). The projection to the MS was threefold stronger than that to the MTL. Moreover, a majority of NI neurons projected independently to either MS or the MTL. However, RLN3 + neurons collateralize significantly more than RLN3-negative (–) neurons. In in vivo studies, electrical stimulation of the NI induced theta activity in the MS and the entorhinal cortex, which was impaired by intraseptal infusion of an RXFP3 antagonist, R3(BΔ23-27)R/I5, particularly at ~ 20 min post-injection. These findings suggest that the MS plays an important relay function in the NI-induced generation of theta within the entorhinal cortex.Funding for open access charge: CRUE-Universitat Jaume IThis research was funded by the Postdoctoral Program of the UJI POSDOC/2021/19 (IG-M); UJI Predoctoral Program PREDOC/2021/19 (MN-S); Fundación Alicia Koplowitz, Spain, grant number 19I436 (FEO-B, FR-B, EC-G); the Spanish Ministerio de Ciencia, Innovación y Universidades, grant number RTI2018-095698-B-I00 (FEO-B, IG-M, FR-B, EC-G); AICO Generalitat Valenciana, grant number AICO/2021/246 (EC-G, FO-B, FR-B), National Health and Medical Research Council of Australia, grant number (ALG), the Spanish Ministerio de Ciencia, Innovación y Universidades, grant number PID2019-107809RB-I00 (AN-M) and Universitat Jaume I, grant numbers UJI-A2017-17 (FR-B) and UJI-B2019-54 (FEO-B).Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. Funding was provided by the Universitat Jaume I, POSDOC/2021/19, PREDOC/2021/19, UJI-A2017-17, Ministerio de Ciencia, Innovación y Universidades, PID2019-107809RB-I00, RTI2018-095698-B-I00, Fundación Alicia Koplowitz, 19I436, Conselleria de Innovación, Universidades, Ciencia y Sociedad Digital, Generalitat Valenciana, AICO/2021/246, National Health and Medical Research Council of Australia, 1067522

The use of GIS and Multi-Criteria evaluation (MCE) to determine scenarios of potential floodings in fluvial basins : study case: cuenca Guanabo, Habana city

El trabajo muestra la potencialidad que presentan los Sistemas de Información Geográfica (SIG) y la evaluación multicriterio (EMC), para determinar con precisión, la cartografía de las zonas susceptibles al desarrollo de inundaciones recurrentes en cuencas fluviales. La capacidad de análisis y manejo de la información espacial que poseen los SIG en la actualidad, conjuntamente con el procesamiento estadístico apoyados en el uso de la EMC, permiten cuantificar factores del medio natural y establecer grados de adecuación, reduciendo la subjetividad durante el procesamiento analítico. Las zonas susceptibles determinadas con el empleo de esta metodología se comparó con estudios anteriores y los resultados presentan una buena coincidencia espacial aunque ésta mejora en precisión cartográfica. Sin duda los resultados obtenidos constituyen valiosas herramientas de gestión, los cuales ayudan a la toma de decisiones en los territorios e incrementan el nivel de conocimiento para reducir los riesgos de desastres naturales.The work shows the potential that Geographic Information Systems and Multi-Criteria Evaluation (EMC) have, to determine accurately, the cartography of favorable areas to recurrent floods development in fluvial basins. The analysis and special information handling capacity that the SIGs have nowadays, together with statistic processing resting in the (EMC) use, make it possible to quantify natural resources factors and establish degrees of fitness, reducing subjectivity during analytic processing. The capable zones determined with the employment of this methodology were compared with previous studies and the result presents a good spatial coincidence though this one improves in cartographic precision. Certainly, the results obtained become valuable management tools that help territory decision making and increase knowledge level to reduce natural disasters risks.Fil: Olivera Acosta, Jorge. La Habana (Cuba). Agencia de medio ambienteFil: Morales, Gustavo Martín. La Habana (Cuba). Agencia de medio ambienteFil: García Rivero, Alberto E.. La Habana (Cuba). Agencia de medio ambienteFil: Salgado, Efrén Jaimez. La Habana (Cuba). Agencia de medio ambienteFil: López, Martin Luis. La Habana (Cuba). Agencia de medio ambienteFil: Estrada, Reinaldo. La Habana (Cuba). Agencia de medio ambienteFil: Campos Dueñas, Mario. La Habana (Cuba). Agencia de medio ambienteFil: Miravet, Bárbara Liz. La Habana (Cuba). Agencia de medio ambient

Solar photovoltaic technology in isolated rural communities in Latin America and the Caribbean

The main characteristics of photovoltaic (PV) energy and its current development in Latin American and Caribbean countries (LAC); its impact on the electrification of homes, health institutions, and schools in isolated or difficult-to-access communities; and, the advantages thereof are presented and discussed by replacing the use of traditional fuels such as firewood and kerosene in order to improve inhabitants ’health as well as reducing CO2 emissions. Countries like Nicaragua, Peru, Brazil, Argentina, and Chile stand out for their growing PV energy development in the region. A case study of the electrification process by PV systems shows very positive changes are manifested in terms of improving the quality of life of the inhabitants, and especially their physical and mental health state. In addition, CO2 emission reductions from electrifying 216 houses in the nine communities reach an annual amount of 2,164.19 t/yr, reducing firewood consumption by 2,123.39 t/yr and kerosene consumption by 40.80 t/yr However, LAC countries must adopt laws and regulations that regulate the use of PV energy, with an emphasis on recycling systems at the end of their life cycle

Determinación de escenarios de inundaciones en la subcuenca del río Cunas, Junín, Perú

Se presenta una metodología basada en el procesamiento del modelo digital del terreno en SAGA-GIS para determinar los escenarios de inundaciones en caso de lluvias intensas o prolongadas en la subcuenca del río Cunas, Junín, Perú. Las variables calculadas a partir del DTM ALOS PALSAR fueron: Depresiones cerradas (CD), Distancia vertical a la red de drenaje (VD), Área de captación modificada (MCA), Escorrentía superficial - Escorrentía terrestre flujo D8, y el índice de humedad topográfico (TW1). La susceptibilidad a las inundaciones se obtiene como CD + MCA + D8 + TW1- VD. El mapa resultante validado en el campo, muestra diferentes grados de susceptibilidad a inundaciones, destacando tres sectores donde un grupo significativo de núcleos de población se encuentran expuestos a potenciales inundaciones de grado medio a alto

Determinación de escenarios de inundaciones en la subcuenca del río Cunas, Junín, Perú

Se presenta una metodología basada en el procesamiento del modelo digital del terreno en SAGA-GIS para determinar los escenarios de inundaciones en caso de lluvias intensas o prolongadas en la subcuenca del río Cunas, Junín, Perú. Las variables calculadas a partir del DTM ALOS PALSAR fueron: Depresiones cerradas (CD), Distancia vertical a la red de drenaje (VD), Área de captación modificada (MCA), Escorrentía superficial - Escorrentía terrestre flujo D8, y el índice de humedad topográfico (TW1). La susceptibilidad a las inundaciones se obtiene como CD + MCA + D8 + TW1- VD. El mapa resultante validado en el campo, muestra diferentes grados de susceptibilidad a inundaciones, destacando tres sectores donde un grupo significativo de núcleos de población se encuentran expuestos a potenciales inundaciones de grado medio a alto

Claiming the patrimonial value of the agrarian. Sugar cane in oriental Malaga

En el ámbito de protección del patrimonio, cada vez existen más autores y trabajos que inciden en la necesidad y la importancia de no concebir aisladamente los testigos inmuebles, muebles y territoriales de una determinada actividad a la hora de aplicarle una protección, pues los unos sin los otros no tienen razón de ser. En la industria de la caña de azúcar esta protección podría en muchos casos incluir el suelo agrario y fuente de materia prima que hizo funcionar dicha industria. De esta forma, la protección tendría efecto por igual sobre los elementos inmuebles (arquitectura), muebles (maquinaria y herramientas) y el territorio (suelo agrario donde se cultivaba la caña que suministraba la fábrica). El trabajo que se presenta a continuación se centra en la importancia de esta coexistencia entre fábrica-maquinariasuelo agrario. El estudio se focaliza en una de las fábricas existentes en el litoral este malagueño cuya situación en la actualidad se torna un tanto confusa debida a la ambigüedad de la norma aplicable. Por este motivo, se investiga la evolución del parcelario agrario en el entorno de la fábrica, así como el marco legislativo que da lugar al grado de protección actual ya que podría verse afectado por diversas actividades proyectadas en el municipio a corto-medio plazo. Se plantea una reflexión sobre la forma de proceder a la hora de proteger el patrimonio agrario en general, y más concretamente aquel vinculado a ciertas industrias que dejaron una profunda huella en el territorio de la Axarquía y sus habitantes.In the field of heritage protection, more and more authors and research emphasize the need and importance of not considering the immovable, movable and territorial testimonies of a certain activity independently when deciding the protection they deserve, since one without the other have no reason to exist. In the sugar cane industry, this protection could in many cases include the agricultural land which was the main source of raw material to such industry. This way, the protection would have an equal effect on the immovable elements (architecture), movable (machinery and tools), and the territory (lands where the canes were grown to supply the factory). The following work stresses on the significance of this coexistence between Factory-machinery- agricultural land. The study focusses on one of the existing factories on the east coast of Malaga, show current situation is somewhat confusing due to the ambiguity of the applicable regulations. For this reason, the evolution of the agricultural land in the area that surrounds the factory is throughtfully studied together with the legislative framework that gives rise to the current degree of protection, since this could be affected by various activities that are planned in the shortmedium term in the town. A reflection on how to proceed when it comes to protecting agricultural heritage in general, and more specifically that linked to certain industries that imprinted a deep agrarian character into the ‘Axarquia’ territory and its inhabitants