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CRISPR-Cas13 RNA-targeting systems are widely used in basic and applied sciences. However, its application has recently generated controversy due to collateral activity in mammalian cells and mouse models. Moreover, its competence could be improved in vivo. Here, we optimized transient formulations as ribonucleoprotein complexes or mRNA-gRNA combinations to enhance the CRISPR-RfxCas13d system in zebrafish. We i) use chemically modified gRNAs to allow more penetrant loss-of-function phenotypes, ii) improve nuclear RNA targeting, and iii) compare different computational models and determine the most accurate to predict gRNA activity in vivo. Furthermore, we demonstrate that transient CRISPR-RfxCas13d can effectively deplete endogenous mRNAs in zebrafish embryos without inducing collateral effects, except when targeting extremely abundant and ectopic RNAs. Finally, we implement alternative RNA-targeting CRISPR-Cas systems such as CRISPR-Cas7-11 and CRISPR-DjCas13d. Altogether, these findings contribute to CRISPR-Cas technology optimization for RNA targeting in zebrafish through transient approaches and assist in the progression of in vivo applications. CRISPR-Cas13 RNA-targeting systems comprise an invaluable set of tools in the fields of basic and applied sciences. Here, Moreno-Sánchez, Hernández-Huertas, and Nahón-Cano et al. enhanced the use of the CRISPR-RfxCas13d system in zebrafish for targeted depletion of endogenous mRNAs.

This study evaluates hindcast simulations performed with a regional climate model (RCM, RegCM4) driven by reanalysis data (ERA-Interim) over the Pacific coast and Andes Cordillera of extratropical South America. A nested domain configuration at \\[0.44^{\\circ }\\] (\\[\\sim\\] 50 km) and \\[0.09^{\\circ }\\] (\\[\\sim\\] 10 km) spatial resolutions is used for the simulations. RegCM4 is also driven by a global climate model (GCM, MPI-ESM-MR) on the same domain configuration to asses the added values for temperature and precipitation (historical simulations). Overall, both 10 km hindcast and historical simulation results are promising and exhibit a better representation of near-surface air temperature and precipitation variability compared to the 50 km simulations. High-resolution simulations suppress an overestimation of precipitation over the Andes Cordillera of northern Chile found with the 50 km simulations. The simulated daily temperature and precipitation extreme indices from 10 km hindcast simulation show a closer estimation of the observed fields. A persistent warm bias (\\[\\sim +\\,{4\\,}^{\\circ }\\hbox {C}\\]) over the Atacama Desert in 10 km hindcast simulation reveals the complexity in representing land surface and radiative processes over the desert. Difficulties in capturing the temperature trend in northern Chile are notable for both hindcast simulations. Both resolutions exhibit added values for temperature and precipitation over large parts of Chile, in particular, the 10 km resolves the coastal-valley Andes transitions over central Chile. Our results highlight that resolutions coarser than 50 km (e.g., GCMs and reanalysis) miss important climate gradients imposed by complex topography. Given that the highest spatial resolution of the current regional simulations over the South America is about 50 km, higher resolutions are important to improve our understanding of the dynamical processes that determine climate over complex terrain and extreme environments.

Air pollution and its effects on population health are currently among the most important public health issues. It is well established that the impact of air pollution on health is exceedingly high, although it ignores its real scope and effects on the aging process because studies on air quality have largely focused on younger age groups. Herein, we emphasize the relevance of air quality to the behavioral aging process, taking into account the place of residence - rural or urban. We raise the following question: Can air quality and residential settings modulate cognitive, emotional and social behaviors during the aging? Some studies have analyzed the role of residential settings and air pollution in the context of a behavioral frame in elderly people. Based on the analyzed literature, this revision concluded that air pollutants affect cognitive function, increasing the risk of dementia as well as depression and anxiety emotional responses. In addition, social networks and inclusion can modulate and mitigate the effects observed during the aging in rural areas that are exposed to less contamination. Although there is no consensus, it seems that some observed behavioral effects are sex-dependent, as women are more vulnerable to air pollution. Additionally, we examined why older adults are vulnerable to the health effects of Particulate Matter (PM) exposure and highlighted the importance of social health in this context. Environmental agents could be the key to understanding the susceptibility and variability observed during aging in behavioral symptoms. Although cognitive decline is related to increased age, it is not a manipulated factor. Efforts should be centered on locating factors implicated in the aging process that could be susceptible to manipulation or variation, such as the choice of the place of residence and the air that we are breathing. Given the significant societal impact of PM, research and policy regulations should be closely aligned and collaborative.

We explored peer acceptance and friendships during adolescence, and their relationship with subsequent changes in the level of adolescents' academic achievement. Participants were 447 students (51% girls) aged from 11 to 16 years. The students completed sociometric assessments of their peer acceptance and friendships during the autumn semester (Time 1). Academic achievement data were also obtained from students' report-card grades at Time 1 as well as during the spring semester (Time 2) of the same academic year. Regression analysis indicated that peer acceptance positively predicted subsequent academic achievement. This relationship was also moderated by age, with the effect of peer acceptance on subsequent academic achievement being greater during early adolescence than in midadolescence. This research deepens understanding of the mechanisms by which positive peer relationships influence adolescents' academic achievement.

The recent pandemic has influenced teachers’ mental health and well-being. The present work follows the Job-Demands-Control model, analyzing changes in the demands, control, and social support during COVID-19, and how they influence job satisfaction, emotional exhaustion, depersonalization, and personal accomplishment among school teachers. The sample comprised 172 school teachers. The instruments applied were The Maslach Burnout Inventory (MBI), The Job Satisfaction Teacher Index (Organization for Economic Cooperation and Development (OECD), 2014), and The Demand Control Support Questionnaire (DCSQ). Job satisfaction was predicted exclusively by social support, revealing the importance of social interactions at work. Emotional exhaustion and depersonalization were predicted by job demands, showing that an increase in job demands (i.e., COVID) affects individuals’ feelings of burnout. The results show that it would be interesting to design strategies that guarantee job control in the teaching context. This would open multiple pathways to implement healthier methodological processes for teachers and the consequent research to support these processes.

CbrAB is a high ranked global regulatory system exclusive of the Pseudomonads that responds to carbon limiting conditions. It has become necessary to define the particular regulon of CbrB and discriminate it from the downstream cascades through other regulatory components. We have performed in vivo binding analysis of CbrB in P. putida and determined that it directly controls the expression of at least 61 genes; 20% involved in regulatory functions, including the previously identified CrcZ and CrcY small regulatory RNAs. The remaining are porines or transporters (20%), metabolic enzymes (16%), activities related to protein translation (5%) and orfs of uncharacterised function (38%). Amongst the later, we have selected the operon PP2810-13 to make an exhaustive analysis of the CbrB binding sequences, together with those of crcZ and crcY. We describe the implication of three independent non-palindromic subsites with a variable spacing in three different targets; CrcZ, CrcY and operon PP2810-13 in the CbrAB activation. CbrB is a quite peculiar σN-dependent activator since it is barely dependent on phosphorylation for transcriptional activation. With the depiction of the precise contacts of CbrB with the DNA, the analysis of the multimerisation status and its dependence on other factors such as RpoN o IHF, we propose a model of transcriptional activation.

The purpose of the study is to comparatively test the expectancy-value model in Chilean and Spanish samples. The model proposes: a social world (composed of social support, physical activity teasing, and weight teasing), expectancy (composed of perceived competence and appearance), task values (composed of enjoyment and stress) to predict physical activity and intention to be physically active. Participants were 497 (Chilean) and 1365 (Spanish) adolescents. Structural equation models and multi-group modelling were used. All the models presented adequate fit to the data. The results show that physical activity teasing is a contextual and essential variable; perceived competence and enjoyment influenced physical activity and intentions to be physically active; some differences appeared in the prediction of physical activity and intentions to be physically active when the multi-group model was run. Culturally tailored interventions are key to improving physical activity (PA) behaviors.

Despite the decline in partially (PM 10 ) and fully (PM 2.5 ) inhalable particles observed in recent decades, Santiago in Chile shows high levels of particle and ozone pollution. Attainment plans have emphasized measures aimed at curbing primary and, to some extent, secondary particles, but little attention has been paid to photochemical pollution. Nevertheless, ozone hourly mixing ratios in Eastern Santiago regularly exceed 110 ppbv in summer, and in winter maximum mixing ratios often reach 90 ppbv. Moreover, the sum of ozone and nitrogen dioxide shows an increasing trend of more than 3.5 ppbv per decade at 5 out of 8 stations. This trend is driven by increasing NO 2 , possibly associated with increasing motorization but also with changes in photochemistry. To estimate the fraction of secondary particles in PM 2.5 and due to the lack of long-term speciation data for particles, we use carbon monoxide as a proxy of primary particles and ozone daily maxima as a proxy for secondary particle formation. We find a growing fraction of secondary particles due to an increase in the oxidizing capacity of Santiago's atmosphere. This stresses the need for new curbing measures to tackle photochemical pollution. This is particularly needed in the context of a changing climate.

Summary Rhodococcus sp. strain TFB is a metabolic versatile bacterium able to grow on naphthalene as the only carbon and energy source. Applying proteomic, genetic and biochemical approaches, we propose in this paper that, at least, three coordinated but independently regulated set of genes are combined to degrade naphthalene in TFB. First, proteins involved in tetralin degradation are also induced by naphthalene and may carry out its conversion to salicylaldehyde. This is the only part of the naphthalene degradation pathway showing glucose catabolite repression. Second, a salicylaldehyde dehydrogenase activity that converts salicylaldehyde to salicylate is detected in naphthalene‐grown cells but not in tetralin‐ or salicylate‐grown cells. Finally, we describe the chromosomally located nag genes, encoding the gentisate pathway for salicylate conversion into fumarate and pyruvate, which are only induced by salicylate and not by naphthalene. This work shows how biodegradation pathways in Rhodococcus sp. strain TFB could be assembled using elements from different pathways mainly because of the laxity of the regulatory systems and the broad specificity of the catabolic enzymes. Three differently regulated sets of enzymes participate in naphthalen degradation in Rhodococcus TFB: the Thn enzymes induced by naphthalene or tetralin but not by salicylate, the salicylaldehyde dehydrogenase induced by naphthalene but not by salicylate or tetralin, and the Nag enzymes induced by salicylate but not by naphthalene itself or tetralin. Therefore, it represents a paradigmatic example of how a biodegradation pathway can be assembled using genes belonging to different regulons.

FixK2 is a CRP/FNR-type transcription factor that plays a central role in a sophisticated regulatory network for the anoxic, microoxic and symbiotic lifestyles of the soybean endosymbiont Bradyrhizobium diazoefficiens. Aside from the balanced expression of the fixK2 gene under microoxic conditions (induced by the two-component regulatory system FixLJ and negatively auto-repressed), FixK2 activity is posttranslationally controlled by proteolysis, and by the oxidation of a singular cysteine residue (C183) near its DNA-binding domain. To simulate the permanent oxidation of FixK2, we replaced C183 for aspartic acid. Purified C183D FixK2 protein showed both low DNA binding and in vitro transcriptional activation from the promoter of the fixNOQP operon, required for respiration under symbiosis. However, in a B. diazoefficiens strain coding for C183D FixK2, expression of a fixNOQP’-‘lacZ fusion was similar to that in the wild type, when both strains were grown microoxically. The C183D FixK2 encoding strain also showed a wild-type phenotype in symbiosis with soybeans, and increased fixK2 gene expression levels and FixK2 protein abundance in cells. These two latter observations, together with the global transcriptional profile of the microoxically cultured C183D FixK2 encoding strain, suggest the existence of a finely tuned regulatory strategy to counterbalance the oxidation-mediated inactivation of FixK2 in vivo.