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The sentinel plantation concept consists of assessing the impact of exotic factors, such as pests and pathogens, on plants of interest by planting them out of their native range. This tool is a way to enhance knowledge for pest risk analysis (PRA) by guiding decisions on how quarantine organisms should be regulated and where to focus prevention and surveillance efforts for an early detection. In this study, the sentinel method was used in the case of research on Xylella fastidiosa, a plant pathogenic bacterium that has recently been found established in southern Europe, but whose potential impact and possible host range are still poorly documented in northern areas where the bacterium is not known to occur. To improve knowledge on the susceptibility of potential hosts of X. fastidiosa in northern Europe, a sentinel plantation of Prunus domestica cv. Opal, Quercus petraea and Salix alba was established in the X. fastidiosa-infected area of Majorca. In order to assess the circulation of the bacterium in the sentinel plot and around it, surveys of the local flora and insect vectors were carried out, as well as the planting of a network of rosemary “spy plants”. Symptomatic monitoring and molecular analyses were performed on the sentinel plants for four years. During these years, X. fastidiosa was never detected in our sentinel plants most likely because of the low infectivity pressure recorded in the surroundings. This study underlines the complexity of conducting sentinel plantation assays combined with X. fastidiosa research, highlighting the need for long-term investigation and questioning the efficiency of the sentinel tool. However, this study is placed in perspective with other valuable sentinel plantations. It also highlights the complementarity of the tool and proposes elements to improve or re-orientate the implementation of future sentinel projects.

Culicoides biting midges (Diptera: Ceratopogonidae) are the main vectors of livestock diseases such as bluetongue (BT) which mainly affect sheep and cattle. In Spain, bluetongue virus (BTV) is transmitted by several Culicoides taxa, including Culicoides imicola, Obsoletus complex, Culicoides newsteadi and Culicoides pulicaris that vary in seasonality and distribution, affecting the distribution and dynamics of BT outbreaks. Path analysis is useful for separating direct and indirect, biotic and abiotic determinants of species' population performance and is ideal for understanding the sensitivity of adult Culicoides dynamics to multiple environmental drivers. Start, end of season and length of overwintering of adult Culicoides were analysed across 329 sites in Spain sampled from 2005 to 2010 during the National Entomosurveillance Program for BTV with path analysis, to determine the direct and indirect effects of land use, climate and host factor variables. Culicoides taxa had species-specific responses to environmental variables. While the seasonality of adult C. imicola was strongly affected by topography, temperature, cover of agro-forestry and sclerophyllous vegetation, rainfall, livestock density, photoperiod in autumn and the abundance of Culicoides females, Obsoletus complex species seasonality was affected by land-use variables such as cover of natural grassland and broad-leaved forest. Culicoides female abundance was the most explanatory variable for the seasonality of C. newsteadi, while C. pulicaris showed that temperature during winter and the photoperiod in November had a strong effect on the start of the season and the length of overwinter period of this species. These results indicate that the seasonal vector-free period (SVFP) in Spain will vary between competent vector taxa and geographic locations, dependent on the different responses of each taxa to environmental conditions.

La gobernanza de la salud pública es un sistema de gestión en el que la cultura organizacional, las normas familiares, la formación y las relaciones sindicales serían determinantes de la adherencia al tratamiento como indicador de la calidad del servicio de salud. Se realizó un estudio no experimental, transversal y exploratorio con una selección probabilística de 104 estudiantes asociados con la infección por el VIH. Los resultados muestran que las relaciones en torno al cuidado de los infectados por el VIH determinan la adherencia al tratamiento (β = 0,702, R = 0,645, R2 = 0,080, R2 ajustado = 0,070; p

The public agenda, which reflects the main themes and topics of discussion between the government and the public, has shifted toward debating the pandemic’s impact on public opinion and government initiatives without considering the findings reported in the literature as indicators of the phenomenon. The objective of the present work was to establish the networks inferred from the relationships between findings reported in the literature from 2019 to 2022 regarding political and social actors in the field of social risk communication. The results warn of the prevalence of collateral axes to mobility as a restrictive effect of confinement policies and distancing of people. The analysis of health policies is recommended to anticipate conflict scenarios between the parties involved.

The identity and location of vocalization pattern generating (VPG) circuits in mammals is debated. Based on physiological experiments, investigators suggested anterior brainstem circuits in the reticular formation, and anatomic evidence suggested the nucleus retroambiguus (NRA) in the posterior brainstem, or combinations of these sites as the putative mammalian VPG. Additionally, vocalization loudness is a critical factor in acoustic communication. However, many of the underlying neuronal mechanisms are still unknown. Here, we evoked calls by stimulation of the periaqueductal gray in anesthetized male rats, performed a large-scale mapping of vocalization-related activity using the activity marker c-fos, and high-density recordings of brainstem circuits using Neuropixels probes. Both c-fosexpression and recording of vocalization-related activity point to a participation of the NRA in vocalization. More important, among our recorded structures, we found that the NRA is the only brainstem area showing a strong correlation between unit activity and call intensity. In addition, we observed functionally diverse patterns of vocalization-related activity in a set of regions around NRA. Dorsal to NRA, we observed activity specific to the beginning and end of vocalizations in the posterior level of the medullary reticular nucleus, dorsal part, whereas medial and lateral to the NRA, we observed activity related to call initiation. No clear vocalization-related activity was observed at anterior brainstem sites. Our findings suggest a set of functionally heterogeneous regions around the NRA contribute to vocal pattern generation in rats.SIGNIFICANCE STATEMENTVocalization patterns are shaped in the mammalian brainstem, but the identity and location of the circuits involved is debated. Additionally, the neuronal mechanisms of vocal intensity control are still unknown. This study consisted of a large-scale mapping of brainstem vocalization circuits based on the activity marker c-fosand high-density recordings with Neuropixels probes. The results confirm the role of nucleus retroambiguus in call production and point to a key role of neurons in this nucleus in loudness control. Dorsal to the nucleus retroambiguus and in the posterior medulla, the authors identify neurons with activity specific to the beginning and end of vocalizations. The results point to specific neural dials for various aspects of rat vocalization control in the posterior brainstem.

Polycomb repressive complex 2 (PRC2) has oncogenic and tumor-suppressive roles in cancer. There is clinical success of targeting this complex in PRC2-dependent cancers, but an unmet therapeutic need exists in PRC2-loss cancer. PRC2-inactivating mutations are a hallmark feature of high-grade malignant peripheral nerve sheath tumor (MPNST), an aggressive sarcoma with poor prognosis and no effective targeted therapy. Through RNAi screening in MPNST, we found that PRC2 inactivation increases sensitivity to genetic or small-molecule inhibition of DNA methyltransferase 1 (DNMT1), which results in enhanced cytotoxicity and antitumor response. Mechanistically, PRC2 inactivation amplifies DNMT inhibitor–mediated expression of retrotransposons, subsequent viral mimicry response, and robust cell death in part through a protein kinase R (PKR)–dependent double-stranded RNA sensor. Collectively, our observations posit DNA methylation as a safeguard against antitumorigenic cell-fate decisions in PRC2-loss cancer to promote cancer pathogenesis, which can be therapeutically exploited by DNMT1-targeted therapy.Significance:PRC2 inactivation drives oncogenesis in various cancers, but therapeutically targeting PRC2 loss has remained challenging. Here we show that PRC2-inactivating mutations set up a tumor context–specific liability for therapeutic intervention via DNMT1 inhibitors, which leads to innate immune signaling mediated by sensing of derepressed retrotransposons and accompanied by enhanced cytotoxicity.See related commentary by Guil and Esteller, p. 2020.This article is highlighted in the In This Issue feature, p. 2007

Supplementary Data from PRC2-Inactivating Mutations in Cancer Enhance Cytotoxic Response to DNMT1-Targeted Therapy via Enhanced Viral Mimicry

The objective of this paper is to review the theoretical, conceptual and empirical frameworks around the perception of risk as a rational, deliberate, planned and systematic process of contingencies and threats. A documentary and retrospective study was carried out with a non-probabilistic selection of sources indexed in international repositories, considering the journal's impact factor and the author's citation index. A model was established based on the evaluative consensus of expert judges in the subjects, although the research design limited these findings to the study scenario, suggesting the extension of the work towards lines of research concerning normative, technological and strategic trust with respect to to civil protection.

Background: The Enhanced Recovery After Surgery (ERAS) care pathways include evidence-based items designed to accelerate recovery after surgery. Interdisciplinarity is one of the key points of ERAS programs. Objective: To prepare a consensus document among the members of the Nutrition Area of the Spanish Society of Endocrinology and Nutrition (SEEN) and the Spanish Group for Multimodal Rehabilitation (GERM), in which the goal is to homogenize the nutritional and metabolic management of patients included in an ERAS program. Methods: 69 specialists in Endocrinology and Nutrition and 85 members of the GERM participated in the project. After a literature review, 79 statements were proposed, divided into 5 sections: 17 of general characteristics, 28 referring to the preoperative period, 4 to the intraoperative, 13 to the perioperative and 17 to the postoperative period. The degree of consensus was determined through a Delphi process of 2 circulations that was ratified by a consistency analysis. Results: Overall, in 61 of the 79 statements there was a consistent agreement, with the degree of consensus being greater among members of the SEEN (64/79) than members of the GERM (59/79). Within the 18 statements where a consistent agreement was not reached, we should highlight some important nutritional strategies such as muscle mass assessment, the start of early oral feeding or pharmaconutrition. Conclusion: Consensus was reached on the vast majority of the nutritional measures and care included in ERAS programs. Due to the lack of agreement on certain key points, it is necessary to continue working closely with both societies to improve the recovery of the surgical patients.

Objetivo: Avaliar a associação entre o estado nutricional e a presença de sintomas gastrointestinais em pacientes oncológicos em tratamento quimioterápico. Método: Estudo transversal e prospectivo aplicado em 54 pacientes com diagnóstico de qualquer neoplasia maligna, em tratamento quimioterápico, com idade ?18 anos, de ambos os gêneros. Os dados foram coletados em um hospital universitário terciário e foram avaliadas medidas antropométricas, dietéticas e os sintomas gastrointestinais foram avaliados pelo questionário Common Terminology Criteria for Adverse Event (CTCAE). Para todas as análises foi fixado um nível de significância de 5%. Resultados: Os pacientes apresentaram média de idade de 59,3±14,5 anos, sendo a maioria idosos (57,4%) e do sexo feminino (55,6%). O IMC alcançou média de 26,2±5,4 kg/m², sendo identificado uma prevalência de 50% de excesso de peso e obesidade, 35,1% de eutrofia e 14,8% de baixo peso. Foi encontrada maior frequência de idosos com baixo peso quando comparado com adultos. Identificou-se alta prevalência de sintomas gastrointestinais sendo os mais relatados: xerostomia (74%), disosmia (61%), disgeusia (61%), saliva espessa ou gosto metálico na boca (59,2%) e náusea (57,3%). Os participantes apresentaram média de consumo energético de 1.588±688 Kcal/dia e ingestão proteica menor do que recomendado na maioria dos pacientes. Indivíduos com excesso de adiposidade e pacientes com consumo energético maior relataram mais sintomas. Conclusão: Apesar da alta prevalência de sintomas gastrointestinais, indivíduos com câncer em tratamento quimioterápico não apresentaram impactos significativos no estado nutricional.

Neuroprosthetic technology aims to restore nervous system functionality in cases of severe damage or degeneration by recording and stimulating the electrical activity of the neural tissue. One of the key factors determining the quality of the neuroprostheses is the electrode material used to establish electrical communication with the neural tissue, which is subject to strict electrical, electrochemical, and mechanical specifications as well as biological and microfabrication compatibility requirements. This work presents a nanoporous graphene-based thin film technology and its engineering to form flexible neural implants. Bench measurements show that the developed microelectrodes offer low impedance and high charge injection capacity throughout millions of pulses. In vivo electrode performance was assessed in rodents both from brain surface and intracortically showing high-fidelity recording performance, while stimulation performance was assessed with an intrafascicular implant that demonstrated low current thresholds and high selectivity for activating subsets of axons within the sciatic nerve. Furthermore, the tissue biocompatibility of the devices was validated by chronic epicortical and intraneural implantation. Overall, this works describes a novel graphene-based thin film microelectrode technology and demonstrates its potential for high-precision neural interfacing in both recording and stimulation applications.

Xylella fastidiosaWells (1987) (Proteobacteria:Xanthomonadaceae) is a xylem pathogen bacterium transmitted by xylem feeder insects that causes several important plant diseases such as Pierce’s disease in grapes or leaf scorch in almond and olives trees. The bacterium was detected in the Balearic Islands in October 2016, including three subspecies:fastidiosa,multiplexandpauca. The major potential vectors described in the Balearics arePhilaenus spumariusL. andNeophilaenus campestrisFallen (1805). In order to interfere the life cycle of vectors, we tested the effect of mechanical control of the plant cover on the most vulnerable phases, such as nymphs and/or newly emerged adults. For this, we selected four organic orchards in Mallorca, three olive and one vineyard plots. Owners of each selected plot conducted mechanical control according to their common procedures and their own machinery, which in general included cut and tillage of the plant cover during March-April. Nymph abundance per surface (30 sampling points/treatment/orchard x 0,25 m2) was measured in each plot in a weekly basis before and after mechanical control. Our results indicated that either tillage and mowing decreased nymphal density ofX. fastidiosavectors in both types of crops. These results contribute to the integrated pest management of vectors by conducting feasible farm-based management of the regular plant cover.Abstract Figure

Polycyclic aromatic hydrocarbons (PAHs) are the most widespread xenobiotic pollutants in water and their abatement usually involves expensive and energy-consuming treatments. In this work, anthracene (AN) was selected as the recalcitrant model of PAHs and its solar light-stimulated heterogeneous photocatalytic abatement in aerated aqueous media was investigated using a new TiO derived thermoplastic nanocomposite in thin film form. The results were also compared with the benchmark TiO photocatalyst in slurry form. Finally, the possible contribution of reactive intermediates such as hydroxyl radical, AN radical cation and singlet oxygen, was investigated by using a hydroxyl radical trap and laser flash photolysis. Based on the obtained results, a feasible mechanism for AN photodegradation, which involves hydroxyl radical as the key oxidizing species is proposed., This research was funded by the University of Catania (Piano di incentivi per la ricerca di Ateneo, PIACERI–Linea 2).

Photosensitized thymine<>thymine (Thy<>Thy) formation and repair can be mediated by carbazole (Cbz). The former occurs from the Cbz triplet excited state via energy transfer, while the latter takes place from the singlet excited state via electron transfer. Here, fundamental insight is provided into the role of the topology and excited state multiplicity, as factors governing the balance between both processes. This has been achieved upon designing and synthesizing different isomers of trifunctional systems containing one Cbz and two Thy units covalently linked to the rigid skeleton of the natural deoxycholic acid. The results shown here prove that the Cbz photosensitized dimerization is not counterbalanced by repair when the latter, instead of operating through-space, has to proceed through-bond., Financial support from the Spanish Government (grant SEV-2016-0683), Generalitat Valenciana (PROMETEO/2017/075 and PROMETEO/2021/077), and Technical University of Valencia (Predoctoral FPI fellowship for P.M.) is gratefully acknowledged. G.S. thanks CTI-CSIC for the use of computational facilities.

We know little about mammalian anemotaxis, wind-sensing. Recently, however, Hartmann and colleagues showed whisker-based anemotaxis in rats. To investigate how whiskers sense airflow, we tracked whisker tips in anesthetized or cadaver rats under no airflow, low airflow and high (fan-blowing) airflow. Whisker tips showed little movement under no airflow conditions and all whisker tips moved during high airflow. Low airflow conditions – most similar to naturally occurring wind stimuli – engaged whisker tips differentially. Most whiskers moved little, the long supraorbital whisker showed maximal displacement and α, A1, β, and γ whiskers also showed movements. The long supraorbital whisker differs from other whiskers in its exposed dorsal position, upward bending, length and thin diameter.Ex vivoextracted long supraorbital whiskers also showed exceptional airflow displacement, suggesting whisker-intrinsic biomechanics mediate the unique airflow-sensitivity. Micro computed tomography revealed that the ring-wulst – the follicle structure receiving the most sensitive afferents – was more complete/ closed in supraorbital and other wind-sensitive whiskers than in non-wind-sensitive whiskers, suggesting specialization of the supraorbital for omni-directional sensing. We localized and targeted the cortical supraorbital whisker representation in simultaneous Neuropixels recordings with D/E-row whisker barrels. Responses to wind-stimuli were stronger in the supraorbital whisker representation than in D/E-row barrel cortex. We assessed the behavioral significance of whiskers in an airflow-sensing paradigm. We observed that rats spontaneously turn towards airflow stimuli in complete darkness. Selective trimming of wind-responsive whiskers diminished airflow turning responses more than trimming of non-wind-responsive whiskers. Lidocaine injections targeted to supraorbital whisker follicles also diminished airflow turning responses compared to control injections. We conclude that supraorbital whiskers act as wind antennae.New and NoteworthyAnimals rely on sensory processing of airflow (anemotaxis) to guide navigation and survival. We examined mechanisms of rat anemotaxis by combining whisker tracking, biomechanical analysis, micro computed tomography of follicle structure, Neuropixels recordings in the barrel field, behavior of airflow turning and whisker interference by trimming and lidocaine injections. This diversity of methods led to a coherent pattern of results. Whiskers greatly differ in their airflow sensitivity and strongly wind-responsive whiskers – in particular long supraorbital whiskers – determine behavioral responses to airflow stimuli in rats.

Contaminants of emerging concern (CECs) constitute a wide class of organic molecules, which are found dissolved in wastewaters. Chemical properties of CECs are as wide as their chemical structures; nevertheless, most of the CECs can be removed from wastewaters using Advanced Oxidation Processes (AOPs). However, there are examples of CECs that have proven recalcitrant to AOPs. In such cases, Advanced Reduction Processes (ARPs) include those techniques able to achieve degradation of contaminants by producing highly reactive reducing radicals. Organic photocatalysts have demonstrated to be an efficient alternative to other types of photocatalysts in AOPs. Moreover, several organic photocatalysts, according to their redox properties, could also act as reductants under appropriate scenarios, a process that has been much less explored than oxidation. As an example, Riboflavin (RF) is a natural water-soluble molecule, well-known as vitamin B2. The photophysical and redox properties of RF have been extensively applied for the oxidative photodegradation of contaminants in water. More recently, the potential of RF has been expanded to the photoreduction of more recalcitrant pollutants. For instance, benzotriazoles UV-stabilizers (BUVSs) constitute a vast family of recalcitrant contaminants, which have been successfully degraded in the presence of a commercially available derivative of RF (2′,3′,4′,5′-tetraacetylated riboflavin, RFTA) and visible light, assisted by an amine, which acts as electron donor to the excited states of RFTA, generating the RFTA.. This species ultimately produces the reduction of the selected BUVSs. Photophysical experiments demonstrated the key role of the radical anion RFTA., and allowed determining the rate constants that were correlated to the observed photodegradations., The authors would like to acknowledge H2020/Marie Skłodowska-Curie Actions under the AQUAlity project (Reference: 765860), Consellería d'Educació, Investigació, Cultura i Esport (PROMETEO/2017/075) and the Spanish Ministry of Science, Innovation and Universities (PID2019–110441RB-C33 financed by MCIN/AEI/10.13039/501100011033) are gratefully acknowledged.

Photosensitization by drugs is directly related with the excited species and the photoinduced processes arising from interaction with UVA light. In this context, the ability of gefitinib (GFT), a tyrosine kinase inhibitor (TKI) used for the treatment of a variety of cancers, to induce phototoxicity and photooxidation of proteins has recently been demonstrated. In principle, photodamage can be generated not only by a given drug but also by its photoactive metabolites that maintain the relevant chromophore. In the present work, a complete study of O-desmorpholinopropyl gefitinib (GFT-MB) has been performed by means of fluorescence and ultrafast transient absorption spectroscopies, in addition to molecular dynamics (MD) simulations. The photobehavior of the GFT-MB metabolite in solution is similar to that of GFT. However, when the drug or its metabolite are in a constrained environment, i.e. within a protein, their behavior and the photoinduced processes that arise from their interaction with UVA light are completely different. For GFT in complex with human serum albumin (HSA), locally excited (LE) singlet states are mainly formed; these species undergo photoinduced electron transfer with Tyr and Trp. By contrast, since GFT-MB is a phenol, excited state proton transfer (ESPT) to form phenolate-like excited species might become an alternative deactivation pathway. As a matter of fact, the protein-bound metabolite exhibits higher fluorescence yields and longer emission wavelengths and lifetimes than GFT@HSA. Ultrafast transient absorption measurements support direct ESPT deprotonation of LE states (rather than ICT), to form phenolate-like species. This is explained by MD simulations, which reveal a close interaction between the phenolic OH group of GFT-MB and Val116 within site 3 (subdomain IB) of HSA. The reported findings are relevant to understand the photosensitizing properties of TKIs and the role of biotransformation in this type of adverse side effects., Financial support from the Spanish Ministry of Science and Innovation (RYC-2015-17737, BEAGAL 18/00211, PID2020-115010RB-I00/AEI/10.13039/501100011033, FPU19/00048, PID2019-105512RB-I00/AEI/10.13039/501100011033), the Xunta de Galicia (ED431C 2021/29), the Centro singular de investigación de Galicia accreditation 2019–2022 (ED431G 2019/03), and the European Regional Development Fund (ERDF) is gratefully acknowledged. All authors are grateful to the Centro de Supercomputación de Galicia (CESGA) for use of the Finis Terrae computer.

The sentinel plantation concept consists of assessing the impact of exotic factors, such as pests and pathogens, on plants of interest by planting them out of their native range. This tool is a way to enhance knowledge for pest risk analysis (PRA) by guiding decisions on how quarantine organisms should be regulated and where to focus prevention and surveillance efforts for an early detection. In this study, the sentinel method was used in the case of research on Xylella fastidiosa, a plant pathogenic bacterium that has recently been found established in southern Europe, but whose potential impact and possible host range are still poorly documented in northern areas where the bacterium is not known to occur. To improve knowledge on the susceptibility of potential hosts of X. fastidiosa in northern Europe, a sentinel plantation of Prunus domestica cv. Opal, Quercus petraea and Salix alba was established in the X. fastidiosa-infected area of Majorca. In order to assess the circulation of the bacterium in the sentinel plot and around it, surveys of the local flora and insect vectors were carried out, as well as the planting of a network of rosemary “spy plants”. Symptomatic monitoring and molecular analyses were performed on the sentinel plants for four years. During these years, X. fastidiosa was never detected in our sentinel plants most likely because of the low infectivity pressure recorded in the surroundings. This study underlines the complexity of conducting sentinel plantation assays combined with X. fastidiosa research, highlighting the need for long-term investigation and questioning the efficiency of the sentinel tool. However, this study is placed in perspective with other valuable sentinel plantations. It also highlights the complementarity of the tool and proposes elements to improve or reorient the implementation of future sentinel projects.

The discovery of three subspecies ofXylella fastidiosain Europe has triggered major attention on the potential spread up North of the bacteria. Assessing the susceptibility of a previously unexposed European flora is a key element that remains widely unknown. Under biosafety facility, we evaluated the susceptibility of Salicaceae such asPopulus tremula, Populus canescens, Salix albaandSalix capreaby mechanically inoculating the KLN59.3 GFP-labelledX. fastidiosaat 22 °C and at 28 °C. Bacterial movement and multiplication in plants were investigated by PCR, real-time PCR, confocal or scanning electron microscopy. Nine months post-inoculation, 100 % of the plants tested positive forX. fastidiosa, with the exception of 57% forP. canescensunder the 22 °C-growing conditions. Bacteria were detected up to 120 cm from the inoculation point forS. alba. They were detected in the roots of all species and were successfully isolated forS. albaandP. tremula. Estimates of average CFU/g of plant tissue per species ranged from 1.5E + 03 to 3.5E + 06, with the lowest figures forP. canescensand the highest forP. tremulatogether with high number of totally obstructed vessels observed by confocal microscopy. The possibility of insect transmission was also evaluated using an experimental set up based on Mallorca Island. There, transmission byP. spumariusof bothX. fastidiosaST1 and ST81 was proven onS. alba. We thus demonstrated that indigenous European Salicaceae such asS. albaorP. tremulaare new potential hosts forX. fastidiosa.

Phenols are regarded as highly toxic chemicals. Their effects are difficult to study in in vitro systems because of their ambiguous fate (degradation, auto-oxidation and volatility). In the course of in vitro studies of a series of redox-cycling phenols, we found evidences of cross-contamination in several in vitro high-throughput test systems, in particular by trimethylbenzene-1, 4-diol/trimethylhydroquinone (TMHQ) and 2,6-di-tertbutyl-4-ethylphenol (DTBEP), and investigated in detail the physicochemical basis for such phenomenon and how to prevent it. TMHQ has fast degradation kinetics followed by significant diffusion rates of the resulting quinone to adjacent wells, other degradation products being able to air-diffuse as well. DTBEP showed lower degradation kinetics, but a higher diffusion rate. In both cases the in vitro toxicity was underestimated because of a decrease in concentration, in addition to cross-contamination to neighbouring wells. We identified four degradation products for TMHQ and five for DTBEP indicating that the current effects measured on cells are not only attributable to the parent phenolic compound. To overcome these drawbacks, we investigated in detail the physicochemical changes occurring in the course of the incubation and made use of gas-permeable and non-permeable plastic seals to prevent it. Diffusion was greatly prevented by the use of both plastic seals, as revealed by GC–MS analysis. Gas non-permeable plastic seals, reduced to a minimum compounds diffusion as well oxidation and did not affect the biological performance of cultured cells. Hence, no toxicological cross-contamination was observed in neighbouring wells, thus allowing a more reliable in vitro assessment of phenol-induced toxicity., The authors wish to acknowledge the support of the European Union’s Horizon 2020 research and innovation 493 program under Grant Agreement No 681002 (EU-ToxRisk). We are indebted to CIBEREHD (ISCIII), for supporting our research program on hepatotoxicity of xenobiotics. L.T. was supported by the Miguel 494 Servet I Program financed by the Institute of Health Carlos III (Plan Estatal de I+D+i 2013-2016) 495 and co-financed by the European Regional Development Fund "A way to achieve Europe" (FEDER) 496 through grant CP16/00097. TMS acknowledges her personal predoctoral grant (FI18/00260, 497 ISCIII, Spain).

European guidelines for testing attractant and repellent efficacy (i.e., Product type 19 [PT19]) have been in revision since 2017. A key topic of discussion is the current approach to evaluating topical repellents. The European Chemical Agency has stated field testing should be avoided because of mosquito-borne disease risks. However, the most common laboratory method, the arm-in-cage (AIC) test, may limit the reliable extrapolation of lab results to field conditions. This study’s main goal was to assess alternative laboratory methods for evaluating topical mosquito repellents that use mosquito landing rates more representative of those in the field. The study took place at three European testing labs using 30 study participants per test and the mosquito, Aedes albopictus (Skuse, 1894, Diptera: Culicidae). In phase 1, a conventional AIC test and a sleeved AIC test were performed. Respectively, the arm area exposed was 600 and 100 cm2, and cage volume was 0.040 and 0.064 m3. Mosquito density was the same for both: 1 female/840 cm3. In phase 2, room-based testing (40 ± 5 mosquitoes in 25–30 m3) was used as a proxy for field testing. The mosquito repellent employed was 15% N,N-diethyl-m-toluamide in ethanol at two doses: 1 and 0.5 g/600 cm2. The protection times measured at each laboratory were analyzed both separately and together using nonparametric (Kruskal–Wallis) test. The two alternatives methods showed to be potential alternatives to the current AIC method recreated field mosquito landing rates and achieved reproducible protection times across laboratories., Graphical Abstract Graphical Abstract

Several pathogens are known to be transmitted by arthropods. One of the most relevant, in economic terms, affecting animals is bluetongue virus. Its known vectors are several species of Culicoides midges. In Europe, the considered main vector species are Culicoides imicola and Culicoides obsoletus, though other species may be implicated at different levels. In the present work, the activity of these vector species between sunset and sunrise from May to November is analyzed according to their captures in a rotator bottle collector adapted to a light trap. Additionally, the Culicoides populations of two areas in Mallorca (Spain) with different characteristics (rural vs. urban) are compared. The results indicated that June is the month of higher abundance of C. obsoletus in our climatic conditions—being active during the first three hours after sunset. Conversely, C. imicola reached the maximum level of captures during October, and captures were more evenly distributed during the night. Collections from the two areas revealed that Culicoides populations were composed by the same species; however, abundance and sex ratio presented marked differences. These results add valuable insight into the ecology of Culicoides and may be used to design more accurate strategies to control diseases associated with Culicoides.

9 Pág., Phlebotomine sand flies are vectors of several diseases of importance for public health, including leishmaniosis, bartonellosis, and sand fly fevers. An entomological survey on blood-feeding Diptera was conducted in June–November 2020–2021 to know the diversity of insect vectors in Mallorca (Balearic Islands, Spain). Among the vectors collected, Phlebotomus (Larroussius) perfiliewi Parrot, 1930 was found being the first record of this species in Spain. Phlebotomus perfiliewi s.l. is one of the main vectors of Leishmania infantum in the Mediterranean Basin and Central Asia. The identification of this species was confirmed by both morphological features and DNA barcoding. Phylogenetic analyses showed that the specimens captured were Ph. perfiliewi s.s. (99.85–100% homologues from Italy and Algeria specimens), with a sequence divergence of 0.17%. The cytochrome c oxidase subunit I gene clearly separates the three species that make up the Ph. perfiliewi species complex. In addition, we also provide a brief discussion about their identification remarks, phylogenetic relationships, and vector status., This study was funded by the Agriculture, Fisheries and Food council through the project num. BIA08/20-2 from the agricultural and fishing guarantee fund (FOGAIBA) of the Government of the Balearic Islands. Samplings were supported by own resources of ZAP-UIB and FOGAIBA. R.G.L. was initially supported by a Margalida Comas contract from the Government of the Balearic Islands and from the European Regional Development Fund (FEDER) (REF- PD/038/2019) and he is currently funded by a Juan de la Cierva 2019 Formación contract (FJC2019-041291-I) from the Ministry of Science and Innovation.

No hay resúmenes disponibles. Investigaciones UCA, Año. 1, Vol. 1, 2019-2020: 380-383.

Polycomb Repressive Complex 2 (PRC2) establishes and maintains di- and tri-methylation at histone 3 at lysine 27 (H3K27me2/3) in the genome and plays oncogenic and tumor suppressor roles in context-dependent cancer pathogenesis. While there is clinical success of therapeutically targeting PRC2 core component, EZH2, in PRC2-dependent cancers (e.g., follicular lymphoma, epithelioid sarcoma), it remains an unmet therapeutic bottleneck in PRC2-inactivated cancer. Biallelic inactivating mutations in PRC2 core components are a hallmark feature of high-grade malignant peripheral nerve sheath tumor (MPNST), an aggressive subtype of sarcoma with poor prognosis and no effective targeted therapeutics. Using a custom RNAi-based drop out screen, we observed that PRC2-inactivation is synthetic lethal with DNA methyltransferase 1 (DNMT1) downregulation; we further observed that small molecule DNMT inhibitors (DNMTis) resulted in enhanced cytotoxicity and antitumor response in PRC2-loss cancer context in vitro and in vivo. Mechanistically, DNMTi-mediated de-repression of retrotransposons (e.g., endogenous retroviral elements (ERVs)/LTR, LINE, SINE) and gene targets is partly restricted by PRC2, which potentially contributes to limited therapeutic activity in PRC2-wild-type (wt) cancer context. In contrast, DNMTi treatment synergizes with PRC2 inactivation and cooperatively amplifies the expression of retrotransposons (e.g., ERV/LTR, LINE, SINE), and subsequent viral mimicry response that promotes robust cell death in part through PKR-dependent double stranded-RNA (dsRNA) sensing. Collectively, our observations posit DNA methylation as a safeguard against anti-tumorigenic cell fate decisions in the context of PRC2-inactivation to promote cancer pathogenesis. Further, they identified a novel targeted therapeutic strategy in PRC2-inactivated MPNST and delineated the PRC2-inactivated cancer context for future preclinical exploration and clinical investigation of DNMT1-targeted therapies in cancer.SIGNIFICANCEPRC2-inactivation drives oncogenesis in various cancers but therapeutically targeting PRC2-loss has remained challenging. Here we show that PRC2 inactivating mutations sets up a tumor context-specific liability for synthetic lethal interaction with genetic and therapeutic inhibition of DNMT1. DNMT1 inhibitor-induced cytotoxicity in PRC2-loss cancer context is accompanied by innate immune signaling signature through PKR-mediated sensing of endogenous retrotransposons. These observations posit a therapeutic window via direct anti-tumor effect by DNMT1 inhibitors in PRC2-loss cancers, and point to potentials to be combined with innovative immunotherapeutic strategies to capitalize on innate immune signaling activation.

Immune checkpoint blockade (ICB) has demonstrated clinical success in “inflamed” tumors with significant T-cell infiltrates, but tumors with an immune-desert tumor microenvironment (TME) fail to benefit. The tumor cell-intrinsic molecular mechanisms of the immune-desert phenotype remain poorly understood. Here, we demonstrate that inactivation of the Polycomb-repressive complex 2 (PRC2) core components, EED or SUZ12, a prevalent genetic event in malignant peripheral nerve sheath tumor (MPNST) and sporadically in other cancer types, drives a context-dependent immune-desert TME. PRC2 inactivation reprograms the chromatin landscape that leads to a cell-autonomous shift from primed baseline signaling-dependent cellular responses (e.g., interferon γ) to PRC2-regulated development and cellular differentiation transcriptional programs. Further, PRC2 inactivation reprograms the TME, leads to diminished tumor immune infiltrates and immune evasion through reduced chemokine production and impaired antigen presentation and T-cell priming, and confers ICB primary resistance through blunted T-cell recruitment in vivo. We demonstrate that strategies that enhancing innate immunity via intratumoral delivery of inactivated modified vaccinia virus Ankara (MVA) leads to increased tumor immune infiltrates and sensitizes PRC2-loss tumors to ICB. Our results provide novel molecular mechanisms of context-dependent dysfunctional epigenetic reprogramming that underline the immune-desert phenotype in MPNST and other cancers with PRC2 inactivation. Importantly, our findings highlight genetic-inactivation of PRC2 as a novel context-dependent ICB therapeutic resistance biomarker in cancer, and caution that therapeutic strategies that non-selectively target PRC2 in the host may lead to undesirable context-dependent immune evasion and ICB resistance in tumors. Our studies also point to intratumoral delivery of immunogenic therapeutic viruses as an initial strategy to modulate the immune-desert TME and capitalize on the clinical benefit of ICB.

(1) To determine probabilities of immediate postoperative new motor deficits after no, reversible, and irreversible motor evoked potentials (MEP) deteriorations and (2) to calculate the same outcome considering whether MEP deteriorations were followed by surgical interventions in the absence of confounding factors.We analyzed MEPs from 513 surgeries. Four-limb MEPs were evoked by transcranial electrical stimulation. Baseline recordings were obtained before skin incision and updated before instrumentation. Motor evoked potentials deteriorations were considered significant whenever they showed a persistent, reversible, or irreversible amplitude decrease of80% of the baseline values.Nine patients showed postoperative new motor deficits. Probabilities of postoperative new motor deficits were null, 2.8%, and 36.8% with no, reversible, and irreversible MEP deteriorations, respectively. The risk of immediate postoperative new motor deficits was significantly lower (P = 0.0002) in reversible MEP compared with irreversible MEP deteriorations. In patients showing reversible/irreversible MEP deteriorations in the absence of confounding factors, surgical interventions compared with nonsurgical interventions significantly decreased the risk of immediate postoperative new motor deficits (P = 0.0216).This study shows that probabilities of immediate postoperative new motor deficits increase with the severity of intraoperative MEP changes. In addition, our results support the value of surgical interventions triggered by MEP deteriorations to reduce postoperative adverse motor outcomes.

Culicoides Latreille (Diptera: Ceratopogonidae) transmit arboviruses affecting wild and domestic ruminants such as bluetongue (BTV) and Schmallenberg virus (SBV). The sub-adult development and lifespan of Culicoides obsoletus s.s. (Meigen), Culicoides circumscriptus Kieffer and Culicoides paolae Boorman were examined at three different temperatures under laboratory conditions. Insects were collected from field between spring and autumn 2015 in two livestock farms located in Majorca (Spain). Gravid females were held individually at 18, 25 or 30 °C. Low temperatures increased the adult lifespan, time to oviposit and rate of development, whereas high temperatures increased the number of eggs, successful pupation and adult emergence as well as the larvae growth rate. The results showed that C. obsoletus s.s. have optimum development at 18 °C, whereas the optimal rearing temperature for C. circumscriptus and C. paolae was under warmer conditions of 25-30 °C. Variations in temperature/humidity and assays with different materials and substrates for oviposition should be considered in future studies. Understanding the requirements of the different species of Culicoides optimizing the results should be of special interest for predicting environmental change effects on these species, in addition to determining the rearing conditions for candidate Culicoides vectors.

Rats have elaborate cognitive capacities for playing Hide & Seek. Playing Hide & Seek strongly engages medial prefrontal cortex and the activity of prefrontal cortex neurons reflects the structure of the game. We wondered if prefrontal neurons would also show a mirroring of play-related neural activity. Specifically, we asked how does the activity in the rat medial prefrontal cortex differ when the animal plays itself versus when it observes others playing. Consistent with our previous work, when the animal plays itself we observed medial prefrontal cortex activity that was sharply locked to game events. Observing play, however, did not lead to a compara-ble activation of rat medial prefrontal cortex. Firing rates during observing play were lower than during real play. The modulation of responses in medial prefrontal cortex by game events was strong during playing Hide & Seek, but weak during observing Hide & Seek. We conclude the rat prefrontal cortex does not mirror play events under our experimental conditions.

This study linked the fermentation index of two varieties of cocoa beans: CCN 51 and ICS 6 with electrical conductivity (EC) of the water where they were immersed using a laboratory conductimeter. Mixtures of 11 fermented cocoa beans and unfermented were performed in order to obtain fermentation index from 0% (unfermented beans) to that reached by the fermented cocoa beans. Immersion times tested for determination of the electrical conductivity were 90 and 180 minutes. The results of electrical conductivity for CCN 51 and ICS 6 were 56.1 ± 12.0 to 1375.0 ± 28.0 mS/cm and 150.0 ± 19.9 to 1199.3 ± 44.5 mS/cm, respectively. Subsequent conductivity values ​​did not represent significant increases over time. The relationship between the fermentation index and electrical conductivity for CCN 51 showed positive trend, achieving a R2 = 63.1%. The following function by regression: EC (CCN-51) = 1196 + 10.83 * (FI) – 0.06 *(FI)2 got a model with significant adjustment to be used to determine its fermentation index.

Presence of diabetes (types 1 and 2) increases the risk of atherosclerotic cardiovascular disease. Despite adequate metabolic control and treatment of vascular risk factors until the goals recommended by the clinical practice guidelines are achieved, residual cardiovascular risk may be very high in some patients with diabetes. Stratifying the vascular risk for each patient as precisely as possible is therefore necessary. Consolidated strategies to improve patient prognosis include aggressive reduction of LDL cholesterol, blood pressure control, achievement of the best HbA1c control possible without inducing hypoglycemia, use of hypoglycemic drugs shown to have cardiovascular benefits, and use of platelet aggregation inhibitors in patients with greater initial risk. Emerging strategies for patients with very high or extreme risk would include use of drugs intended to decrease triglyceride-rich lipoproteins and inflammation.

Photosensitised biphotonic irradiation of DNA has been rarely addressed, probably due to the difficulties in the experimental design. This is associated with the selection of nucleobases and sensitisers with appropriate absorption spectra and photochemical reactivity, in combination with a laser source emitting intense UVA light of the adequate wavelength. The present paper presents a new strategy involving absorption of a first UVA photon by an adequate sensitiser followed by triplet energy transfer to a pyrimidine (Pyr) derivative and absorption of a second UVA photon by the resulting Pyr triplet excited state. The feasibility of the proposed strategy has been demonstrated using two model reactions: (i) the Norrish-Yang photocyclisation of a tert-butyluracil and (ii) the photohydration of its uracil analogue, lacking the tert-butyl substituent., Financial support by the Spanish Government (O. R-A. for FPU14/05294) and Generalitat Valenciana (PROMETEO/2017/075) is gratefully acknowledged. We also thank Dr Lhiaubet-Vallet for her help with the UPLC-MS/MS experiments.

Bi-stable perception is a strong instance of cognitive self-organization, providing a research model for how ‘the brain makes up its mind.’ The complexity of perceptual bistability prevents a simple attribution of functions to areas, because many cognitive processes, recruiting multiple brain regions, are simultaneously involved. The functional magnetic resonance imaging (fMRI) evidence suggests the activation of a large network of distant brain areas. Concurrently, electroencephalographic and magnetoencephalographic (MEEG) literature shows sub second oscillatory activity and phase synchrony on several frequency bands. Strongly represented are beta and gamma bands, often associated with neural/cognitive integration processes. The spatial extension and short duration of brain activities suggests the need for a fast, large-scale neural coordination mechanism. To address the range of temporo-spatial scales involved, we systematize the current knowledge from mathematical models, cognitive sciences and neuroscience at large, from single-cell- to system-level research, including evidence from human and non-human primates. Surprisingly, despite evidence spanning through different organization levels, models, and experimental approaches, the scarcity of integrative studies is evident. In a final section of the review we dwell on the reasons behind such scarcity and on the need of integration in order to achieve a real understanding of the complexities underlying bi-stable perception processes.

EFSA received a mandate from the European Commission to assess the effectiveness of some of the control measures against diseases included in the Category A list according to Regulation (EU) 2016/429 on transmissible animal diseases ('Animal Health Law’). This opinion belongs to a series of opinions where these control measures will be assessed, with this opinion covering the assessment of control measures for glanders. In this opinion, EFSA and the AHAW Panel of experts review the effectiveness of: (i) clinical and laboratory sampling procedures, (ii) monitoring period and (iii) the minimum radius of the protection and surveillance zone, and the minimum length of time the measures should be applied in these zones. The general methodology used for this series of opinions has been published elsewhere. Considering the epidemiology and distribution of glanders, it was foreseen that three different situations could lead to a suspicion of the disease. Sampling procedures were defined for each of the three different suspicion types, which can also be applied in most of the other scenarios assessed. The monitoring period (6 months) was assessed as effective in all scenarios. The AHAW Panel of experts considered the minimum radius and duration of the existing protection and surveillance zone, set at the establishment level, effective. Recommendations provided for each of the scenarios assessed aim to support the European Commission in the drafting of further pieces of legislation, as well as for plausible ad hoc requests in relation to glanders.

Photorepair mechanism of (6-4) photoproducts (6-4PP) by photolyase has been the subject of active debate over the years. The initial rationalization based on electron transfer to an oxetane or azetidine intermediate formed upon binding to the enzyme has been questioned, and there is now a more general consensus that the lesion is directly reduced from the excited flavin cofactor. However, the accepting moiety, i.e. the 5-methyl-2-pyrimidone or 5-hydroxy-5,6-dihydrothymine, has not been fully identified yet. In this work, spectroscopic experiments have been run to determine which of the 5′- or 3′-base of 6-4PP is more prone to be reduced. For this aim, the two building blocks of 6-4PP were synthesized and used as electron acceptors. Instead of the short-lived photolyase cofactor, which does not provide a time window compatible with diffusion-controlled intermolecular processes, carbazole, 2-methoxynaphthalene and phenanthrene have been selected as electron donors due to their appropriate singlet lifetimes and reduction potentials. Steady-state and time-resolved fluorescence revealed that, in solution, the pyrimidone chromophore is the most easily reduced moiety. This was confirmed by transient absorption experiments consisting of quenching of the solvated electron by the two moieties of 6-4PP., This work has been supported in part by the project PGC2018-096684-B-I00 funded by Spanish Government MCIN/AEI/10.13039/501100011033/ and FEDER “Una manera de hacer Europa” and the Generalitat Valenciana (Prometeo/2017/075).

Hepatitis C, a liver inflammation caused by the hepatitis C virus (HCV), is treated with antiviral drugs. In this context, simeprevir (SIM) is an NS3/4A protease inhibitor used in HCV genotypes 1 and 4. It is orally administered and achieves high virological cure rates. Among adverse reactions associated with SIM treatment, photosensitivity reactions have been reported. In the present work, it is clearly shown that SIM is markedly phototoxic, according to the in vitro NRU assay using BALB/c 3T3 mouse fibroblast. This result sheds light on the nature of the photosensitivity reactions induced by SIM in HCV patients, suggesting that porphyrin elevation in patients treated with SIM may not be the only mechanism responsible for SIM-associated photosensitivity. Moreover, lipid photoperoxidation and protein photooxidation assays, using human skin fibroblasts (FSK) and human serum albumin (HSA), respectively, reveal the capability of this drug to promote photodamage to cellular membranes. Also, DNA photo lesions induced by SIM are noticed through comet assay in FSK cells. Photochemical and photobiological studies on the mechanism of SIM-mediated photodamage to biomolecules indicate that the key transient species generated upon SIM irradiation is the triplet excited state. This species is efficiently quenched by oxygen giving rise to singlet oxygen, which is responsible for the oxidation of lipids and DNA (Type II mechanism). In the presence of HSA, the photobehavior is dominated by binding to site 3 of the protein, to give a stable SIM@HSA complex. Inside the complex, quenching of the triplet excited state is less efficient, which results in a longer triplet lifetime and in a decreased singlet oxygen formation. Hence, SIM-mediated photooxidation of the protein is better explained through a radical (Type I) mechanism., Financial support from the Spanish Ministry of Science and Innovation [PID2020-115010RB-I00/AEI/10.13039/501100011033 (I.A), PID2019-105512RB-I00/AEI/10.13039/501100011033 (CG-B) and FPU predoctoral fellowship for M. El O.], Axencia Galega de Innovaci´on (2020-PG067, CG-B), the Xunta de Galicia [ED431C 2021/29 and the Centro singular de investigaci´on de Galicia accreditation 2019–2022 (ED431G 2019/03),], and the European Regional Development Fund (ERDF) is gratefully acknowledged. All authors are grateful to the Centro de Supercomputaci´on de Galicia (CESGA) for use of the Finis Terrae computer.

Joint femtosecond fluorescence upconversion experiments and theoretical calculations provide a hitherto unattained degree of characterization and understanding of the mutagenic etheno adduct 3,N4-etheno-2′-deoxycytidine (ϵdC) excited state relaxation. This endogenously formed lesion is attracting great interest because of its ubiquity in human tissues and its highly mutagenic properties. The ϵdC fluorescence is modified with respect to that of the canonical base dC, with a 3-fold increased lifetime and quantum yield at neutral pH. This behavior is amplified upon protonation of the etheno ring (ϵdCH+). Quantum mechanical calculations show that the lowest energy state ππ*1 is responsible for the fluorescence and that the main nonradiative decay pathway to the ground state goes through an ethene-like conical intersection, involving the out-of-plane motion of the C5 and C6 substituents. This conical intersection is lower in energy than the ππ∗ state (ππ*1) minimum, but a sizable energy barrier explains the increase of ϵdC and ϵdCH+ fluorescence lifetimes with respect to that of dC., Support from the Spanish (project PGC2018-096684-B-I00 funded by MCIN/ AEI /10.13039/501100011033/ and FEDER, and grant BES-2016-077517 for P. L.-A.) and regional (Prometeo/2017/075) governments. The research leading to these results has received funding from LASERLAB-EUROPE (grant agreement no. 654148, European Union’s Horizon 2020 research and innovation programme). L.M.F. thanks the PID2019-110091GB-I00 project funded by MCIN/AEI/10.13039/501100011033 for financial support and the Centro de Computación Científica UAM (CCC-UAM) for computing time.

Benzotriazole UV-stabilizers (BUVSs) are commonly used in industry as solar filters, due to their strong UV light absorption. Because of their extended usage, environmental contamination of waters due to BUVSs constitutes a growing concern. Direct photodegradation of BUVSs is not efficient due to their intrinsic thermal pathways to release the absorbed light. Nevertheless, their abatement in natural environments could be assisted by chromophoric dissolved organic matter. Among the BUVSs, three representative candidates were selected, UV-326, UV-327 and UV-328, to demonstrate the potential of Riboflavin (RF) as a natural visible-light absorbing photocatalyst for the abatement of these recalcitrant pollutants under reductive conditions. The use of visible light and DABCO, as a model sacrificial electron donor, generates the radical anion RFTA. This key species reacts with the solar filters, achieving their reductive abatement from the medium. Moreover, the participation of every potential reactive species has been investigated by photophysical techniques, together with determination of the quenching rate constant for every reaction pathway. Consequently, evidence supported the main role of the reductive photodegradation pathway, being RFTA the key species in the abatement of BUVSs., This research was funded by H2020/Marie Skłodowska-Curie Actions under the AQUAlity project (Reference: 765860) and European Union's Horizon 2020 Research and Innovation Program under European Research Council (ERC) through the HyMAP project (grant agreement No. 648319). Project PID2019-110441RB-C33 financed by MCIN/AEI/10.13039/501100011033 and AEI-MICINN/FEDER, UE through the Nympha Project (PID2019-106315RB-I00), the regional government of “Comunidad de Madrid” and the European Structural Funds through their financial support to FotoArt-CM project (S2018/NMT-4367) are also acknowledged.