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OBJECTIVE: The present study sought to evaluate the psychometric properties and inter-informant agreement of the Self-Injurious Thoughts and Behaviors Interview-Revised (SITBI-R) in adolescents and their parents. METHOD: Suicidal and nonsuicidal adolescents from the community (N =206), ages 12–19 years, were administered the SITBI-R during a lab visit. Approximately half of the adolescents’ parents opted to complete a parent assessment, including the SITBI-R, on behalf of their child. Inter-rater reliability, convergent validity, and inter-informant agreement were assessed. RESULTS: The SITBI-R exhibited overall excellent inter-rater reliability and good convergent validity in adolescents. Parent-adolescent agreement ranged from fair to poor across most outcomes, with parents tending not to report past self-injurious thoughts and behaviors endorsed by their children. CONCLUSIONS: The present study suggests that the SITBI-R can be extended for use with adolescents. Future research should evaluate moderators and implications of parent-adolescent disagreement, particularly with regard to suicide risk.

The study of phytotherapy in dentistry is relevant due to the low occurrence of research in the treatment of oral pathologies, such as caries and periodontal disease. Thus, the aim of this research was to characterize the chemical composition of extracts from the leaves of Couroupita guianensis Aubl, including toxicological evaluation and the antioxidant and antimicrobial potential against Staphylococcus aureus, Streptococcus mutans and Candida albicans. Three extracts were prepared using assisted ultrasound, originating the Crude Ultrasound Extract (CUE) and Soxhlet apparatus, originating the Crude Soxhlet Extract (CSE) and the Ethanol Soxhlet Extract (ESE). In the analysis of the chemical composition, the presence of flavonoids, tannins and saponins were detected. LC-DAD analysis revealed the presence of caffeic acid, sinapic acid, rutin, quercetin, luteolin, kaempferol and apigenin in all extracts. In the GC-MS analysis, stigmasterol and β-sitosterol were identified in CUE and CSE. The antioxidant potentials, determined by the DPPH• and ABTS•+ methods, the ESE showed higher antioxidant activity (2.98 ± 0.96 and 4.93 ± 0.90, respectively). In the evaluation of toxicity, the CUE 50µg/mL and the ESE 50 µg/mL presented growth stimulation of Allium cepa roots. At a concentration of 750µg/mL, all extracts inhibited root growth. None of the extracts showed toxicity against Artemia salina. Antibacterial action was detected in all extracts, mainly against the microorganisms S. aureus and S. mutans; however, the antifungal action against C. albicans was not detected. From the results, the extracts of C. guianensis have therapeutic potential for use in the control of microorganisms in the oral microbiota.

Aroma has a major role in coffee acceptance by consumers. This work has assessed the application of cold plasma technology to induce modifications in ground coffee beans without resorting to solvents or additives, thus applying a green chemistry approach. Cold plasma improved and modulated coffee aroma, increasing desirable and decreasing undesirable aroma notes. Ground coffee beans were subjected to dielectric-barrier discharge (DBD) plasma at different excitation frequencies. Its volatile chemical profile was determined by GC-MS. The odor activity value technique was used to evaluate the aroma profile. Extensive chemical changes occurred with aldehydes, furans, and pyrazine compounds. This work showed that setting proper excitation frequency enables modulating changes toward oxidation and C-C bond scission. Chemical reaction pathways were postulated and discussed. DBD plasma technology enhanced the importance of the desirable cocoa note from 74.4 to 88.6% and decreased the significance of the green and fruity notes from 8.5 to 3.2% and 2.7 to 1.9%, respectively. Plasma-induced reactions can help mitigating aroma defects.

The increasing contamination of water resources is a severe environmental problem. Manganese is highly soluble in water, has adverse effects on the environment and human health during excessive exposure, and is difficult to remove. Biosorbents have been suggested to remove metals from aqueous solutions, therefore, apple pomace was utilized to prepare activated biochar (activated charcoal from apple pomace, ACAP). The biochar sample was characterized by X-ray fluorescence spectrometry, scanning electron microscopy, thermogravimetric analysis, Fourier transform infrared spectroscopy, and Brunauer–Emmett–Teller analysis. The aim of this study was to use ACAP as an adsorbent to remove Mn2+ ions from aqueous solutions. Adsorption experiments were performed in the pH range 3.2–6.5, with various masses of ACAP biochar and Mn2+ ion concentrations. The results were evaluated using the pseudo-first order, pseudo-second order, general order, Elovich, and Webber and Morris kinetics models. All the models fitted the experimental results, the equilibrium time was 15 min, and the pseudo-second order model presented the highest constant (k2), indicating that chemosorption is an important mechanism in adsorption. The adsorption isotherm results were adjusted using different models, wherein the Langmuir and Temkin models exhibited the best fit to the experimental results. Adsorption experiments yielded a 97.5% removal rate of Mn2+ ions at pH 6.5 and 2+ ions in aqueous media, and is applicable for treating water for human consumption.

1H spin-lattice relaxation time (T1) measurements were performed to probe the dynamic behavior of water in aqueous suspensions of cellulose nanocrystals (CNCs) and a layered smectite clay mineral with different degrees of concentration. 1H-T1 experiments were carried out over a wide frequency domain, ranging from a few kilohertz to 500 MHz, with the aid of conventional and fast field cycling nuclear magnetic resonance (NMR) techniques. The experimental relaxometry data illustrate differences between the dynamic behavior of bulk water and that confined in the vicinity of CNC-clay surfaces. Clay alone in moderate concentration was found to enforce almost no effect on the water dynamics, whereas introducing CNCs to the system presented a significantly enhanced relaxivity. The modeling of the relaxation dispersions allowed the determination of dynamical processes and variables explaining the dynamic behavior of water in CNC-clay suspensions. It turned out that reorientations mediated by translational displacements are a leading NMR relaxation mechanism for water interacting with the surfaces of CNC-clay particles in the low-frequency domain. In the high-frequency regime, however, the inner-sphere paramagnetic relaxation mechanism dominates, which is caused by the interaction of water protons with dissolved Fe ions.

AG and TA acknowledge the funding from Slovenian Research Agency, Basic core funding grant P1-0125. SNF, MGH acknowledge the FCT—Portuguese Foundation for Science and Technology projects PIDDAC (POCI-01-0145-FEDER-007688, Reference UIDB/50025/2020-2023). Publisher Copyright: © 2022, The Author(s). Local molecular ordering in liquids has attracted a lot of interest from researchers investigating crystallization, but is still poorly understood on the molecular scale. Classical nucleation theory (CNT), a macroscopic thermodynamic description of condensation, has shortcomings when dealing with clusters consisting of tens of molecules. Cluster formation and local order fluctuations in liquid media are difficult to study due to the limited spatial resolution of electron- and photon-imaging methods. We used NMR relaxometry to demonstrate the existence of dynamic clusters with short-range orientational order in nominally isotropic liquids consisting of elongated molecules. We observed clusters in liquids where the local ordering is driven by polar, steric, and hydrogen-bond interactions between the molecules. In the case of a liquid crystal, measuring the local orientational order fluctuations allowed us to observe the size of these clusters diverging when approaching the phase transition from the isotropic to the nematic phase. These fluctuations are described in terms of rotational elasticity as a consequence of the correlated reorientations of the neighbouring molecules. Our quantitative observations of the dynamic clusters in liquids, numbering about ten or fewer molecules, indicate that this is a general phenomenon in various types of liquids. publishersversion published

Insect-specific viruses (ISVs) are viruses that replicate exclusively in arthropod cells. Many ISVs have been studied in mosquitoes as many of them act as vectors for human etiological agents, such as arboviruses. Aedes (Stegomyia) albopictus is an important potential vector of several arboviruses in Brazil, such as dengue (DENV), Zika (ZIKV) and chikungunya (CHIKV). The development of next-generation sequencing metagenomics has enabled the discovery and characterization of new ISVs. Ae. albopictus eggs were collected using oviposition traps placed in two urban parks in the city of São Paulo, Brazil. The Aedes albopictus females were divided into pools and the genetic material was extracted and processed for sequencing by metagenomics. Complete genomes of ISV Wenzhou sobemo-like virus 4 (WSLV4) were obtained in three of the four pools tested. This is the first detection of ISV WSLV4 in Ae. albopictus females in Latin America. Further studies on ISVs in Ae. albopictus are needed to better understand the role of this species in the dynamics of arbovirus transmission in the Americas.

Insect-specific viruses (ISVs) are viruses that replicate exclusively in arthropod cells. Many ISVs have been studied in mosquitoes as many of them act as vectors for human etiological agents, such as arboviruses.

Cellulose-based systems are useful for many applications. However, the issue of self-organization under non-equilibrium conditions, which is ubiquitous in living matter, has scarcely been addressed in cellulose-based materials. Here, we show that quasi-2D preparations of a lyotropic cellulose-based cholesteric mesophase display travelling colourful patterns, which are generated by a chemical reaction-diffusion mechanism being simultaneous with the evaporation of solvents at the boundaries. These patterns involve spatial and temporal variation in the amplitude and sign of the helix´s pitch. We propose a simple model, based on a reaction-diffusion mechanism, which simulates the observed spatiotemporal colour behaviour. Cellulose is a naturally occurring system of interest for its chiral macromolecular structure. Here, colourful structural patterns are observed in a cellulose-based lyotropic cholesteric system, attributed to conformational self-adjustment of the cellulose chain to its environment.

Overbank flows occur in alluvial valleys during flood events when the conveyance of main channel of rivers is exceeded. Once floodplains are inundated and the so-called compound channel flow is observed, the faster flow in the main channel interacts with the slower flow in the floodplain featuring a much more pronounced 3D flow structure compared to single channel flow. These flow mechanisms comprise a shear layer near the interface, lateral momentum transfer and strong secondary currents due to the non-isotropic turbulence. This paper starts by giving an overview of the main flow mechanisms in compound channels pointing out the importance of taking into account the apparent shear stress generated between the main channel and the floodplain flows due to the interaction of these flows. A new simple model was developed to include the apparent shear stress concept as a correction of the Manning roughness coefficient of main channel and floodplains. The proposed method for predicting stage–discharge relationships was calibrated and validated by experimental data from several compound channel facilities. A significant improvement in prediction of the compound channel conveyance in comparison with the traditional methods was achieved.

Sleep disturbances have been linked to suicidal ideation and behaviors in adolescents. Specifically, insomnia and nightmares are associated with current suicide risk and predict future ideation. Associations between hypersomnia, sleep apnea, and suicide remain inconclusive. Potential biological mechanisms underlying these relationships include executive functioning deficits and hyperarousal. Related psychological factors may include thwarted belongingness, perceived burdensomeness, and negative appraisals. Assessing suicide risk in patients with sleep disturbances, and vice versa, is needed. Therapeutic interventions such as cognitive behavior therapy for insomnia and imagery rehearsal treatment, as well as pharmacologic treatments, show promise in treating sleep disorders and suicidal behavior.

CRUE-CSIC agreement; Springer Nature; Agencia Estatal de Investigación, Spain [PCIN-2016-120, PCIN-2016-063]; Fundacao para a Ciencia e Tecnologia, Portugal [BIODIVERSA/0005/2015, BIODIVERSA/0006/2015]; Agence Nationale de la Recherche, France [ANR-16-EBI3-0006-01, ANR-16-EBI3-0006-02]; AA is supported by a FPU fellowship (Ref. FPU2016-04258, Spanish Ministry of Science, Innovation and Universities). KG is supported by a fellowship from the Severo Ochoa PhD program (Ref. PA-18-PF-BP17-184, Principado de Asturias). GM is supported by post-doctoral contracts from projects PERCEBES (PCIN-2016-063) and MARISCO (CTM2014-51935-R, Spanish Ministerio de Economía y Competitividad)., Aguión, A., Cruz, T., Acuña, J.L., Broudin, C., Castro, J.J., Davoult, D., Dubert, J., Fernandes, J.N., Geiger, K.J., Jacinto, D., Mateus, D., Muñiz, C., Nolasco, R., Perrier, L., Queiroga, H., Román, S., Silva, T., Thiébaut, E., Vázquez, E., Macho, G.

The stalked barnacle Pollicipes pollicipes inhabits rocky shores from the Atlantic coasts of Brittany (France) to Senegal. Because of the culinary traditions of southern Europe, stalked barnacles represent an important target species for local fisheries on the Iberian Peninsula. To manage this fishery sustainably, it is therefore important to assess the dynamics of local populations over the Iberian coast, and how they are interconnected at a wider scale using finely tuned genetic markers. In this work, a new enriched library of GT microsatellites for P. pollicipes was prepared and sequenced using Ion Torrent™ Next Gen-Sequencing Technology. 1,423 adults and juveniles were sampled in 15 localities of three geographic regions: southern Portugal, Galicia and Asturias (both in northern Spain). Twenty polymorphic loci arranged in five multiplex PCRs were then tested and validated as new molecular tools to address the spatial and temporal genetic patterns of P. pollicipes. Our results revealed high genetic diversity among adults. However, juveniles were genetically more structured than their adult counterparts, which alternatively displayed much more connectivity among the three studied regions. The lack of spatial genetic heterogeneity in adults may be due to the overlapping of several generations of settlers coming from different geographic origins, which mainly depends on the orientation of residual currents along the coast during reproduction. The genetic differentiation of juveniles may indeed be congruent with Iberian Peninsula hydrodynamics, which can produce chaotic genetic patchiness (CGP) at small temporal scales due to sweepstake reproductive success, collective dispersal and/or self-recruitment. Remarkably, most of the genetic heterogeneity of juveniles found in this work was located in Galicia, which could represent an admixture between distinct metapopulations or an old refuge for the most northern populations. To conclude, high genetic variation in P. pollicipes can lead to the false impression of population panmixia at the Iberian scale by masking more restricted and current-driven larval exchanges between regions. This possibility should be taken into consideration for further specific management and conservation plans for the species over the Iberian Peninsula. Ministerio de Economía, Industria y Competitividad | Ref. PCIN-2016-120 Ministerio de Economía, Industria y Competitividad | Ref. PCIN-2016-063 Agencia Estatal de Investigación | Ref. PID2019-108481RB-I00 Fundação para a Ciência e Tecnologia | Ref. UIDB/04292/2020 Xunta de Galicia | Ref. ED431C 2020-05 Ministerio de Ciencia, Innovación y Universidades | Ref. FPU2016-04258 Principado de Asturias | Ref. PA-18-PF-BP17-184 Ministerio de Economía y Competitividad | Ref. CTM2014-51935-R NASA | Ref. 80NSSC20K0074

The eco-evolutionary history of penguins is characterised by shifting from temperate to cold environments. Breeding in Antarctica, the Emperor penguin appears as an extreme outcome of this process, with unique features related to insulation, heat production and energy management. However, whether this species actually diverged from a less cold-adapted ancestor, more ecologically similar to its sister species, the King penguin, is still an open question. As the Antarctic colonisation likely resulted in vast changes in selective pressure experienced by the Emperor penguin, the relative quantification of the genomic signatures of selection, unique to each sister species, could answer this question. Applying phylogeny-based selection tests on 7651 orthologous genes, we identified a more pervasive selection shift in the Emperor penguin than in the King penguin, supporting the hypothesis that its extreme cold adaptation is a derived state. Furthermore, among candidate genes under selection, four (TRPM8, LEPR, CRB1, and SFI1) were identified before in other cold-adapted homeotherms, like the woolly Mammoth, while other 161 genes can be assigned to biological functions relevant to cold adaptation identified in previous studies. Location and structural effects of TRPM8 substitutions in Emperor and King penguin lineages support their functional role with putative divergent effects on thermal adaptation. We conclude that extreme cold adaptation in the Emperor penguin largely involved unique genetic options which, however, affect metabolic and physiological traits common to other cold-adapted homeotherms.

The eco-evolutionary history of penguins is profoundly influenced by their shift from temperate to cold environments. Breeding only in Antarctica during the winter, the Emperor penguin appears as an extreme outcome of this process, with unique features related to insulation, heat production and energy management. However, whether this species actually diverged from a less cold-adapted ancestor, thus more similar in ecology to its sister species, the King penguin, is still an open question. As the Antarctic niche shift likely resulted in vast changes in selective pressure experienced by the Emperor penguin, the identification and relative quantification of the genomic signatures of selection, unique to each of these sister species, could answer this question. Applying a suite of phylogeny-based methods on 7,651 orthologous gene alignments of seven penguins and 13 other birds, we identified a set of candidate genes showing significantly different selection regimes either in the Emperor or in the King penguin lineage. Our comparative approach unveils a more pervasive selection shift in the Emperor penguin, supporting the hypothesis that its extreme cold adaptation is a derived state from a more King penguin-like ecology. Among the candidate genes under selection in the Emperor penguin, four genes (TRPM8, LEPR, CRB1, and SFI1) were identified before in other cold adapted vertebrates, while, on the other hand, 161 genes can be assigned to functional pathways relevant to cold adaptation (e.g., cardiovascular system, lipid, fatty acid and glucose metabolism, insulation, etc.). Our results show that extreme cold adaptation in the Emperor penguin largely involved unique genetic options which, however, affect metabolic and physiological traits common to other cold-adapted homeotherms.

Juice processing by non-thermal technology has been extensively studied, aiming at microbial inactivation and quality improvement. However, the knowledge about the possible toxic effects that those technologies can produce in foodstuffs due to the production of reactive oxygen species is still unknown. In this study, sapota-do-Solimões juice processed by ultrasound (2, 6, and 10 min) was evaluated by a toxicity test and protective effect through stress biomarkers (catalase, superoxide dismutase, and lipid peroxidation) using Artemia salina nauplii. The non-thermal processed juice was nontoxic to A. salina. However, the juice fibers imparted some damage to the animal’s body. The ultrasound-processed juice (2 and 6 min) decreased the A. salina mortality to 30% compared to the control assay with H2O2 where mortality was 80% after 48 h of exposure. However, after 72 h of exposure, the A. salina was entirely degraded by H2O2-induced toxicity. Furthermore, the catalase and superoxide dismutase presented the highest activity after A. salina was exposed to the unprocessed juice. Thus, sapota-do-Solimões juice processed by the ultrasound could promote a protective effect on A. salina, revealing this technology’s potential to enhance juice features without toxicity.

Background To investigate the longitudinal relationship between physical frailty, the clinical representation of accelerated biological aging, and antidepressant medication response in older adults with depressive illness. Methods An 8-week randomized placebo-controlled trial (escitalopram or duloxetine) followed by 10 months of open antidepressant medication treatment (augmentation, switch strategies) was conducted in an outpatient research clinic. 121 adults aged 60 years or older with major depressive disorder (MDD) or persistent depressive disorder and a 24-item Hamilton Rating Scale for Depression (HRSD) ≥16 were enrolled. Primary measures assessed serially over 12 months include response (50% reduction from baseline HRSD score), remission (HRSD score Results A 2-class model best fit the data, identifying a consistently low frailty risk (63% of the sample) and consistently high frailty risk (37% of the sample) trajectory. Response and remission rates (ps ≤ .002) for adults in the high-risk frailty class were at least 21 percentage points worse than those in the low-risk class over 12 months. Furthermore, subsequent frailty was associated with previous frailty (ps ≤ .01) but not previous response or remission (ps ≥ .10). Conclusions Antidepressant medication is poorly effective for MDD occurring in the context of frailty in older adults. Furthermore, even when an antidepressant response is achieved, this response does little to improve their frailty. These data suggest that standard psychiatric assessment of depressed older adults should include frailty measures and that novel therapeutic strategies to address comorbid frailty and depression are needed.

In the present work a study of the situation that is presented in Brazil with regard to the sexually transmitted diseases is made, indicating those groups of people for which the most common certain diseases. It introduces a system of differential equations that simulates the process of transmission of different diseases, if different cases are studied that derive from the general system; the system is simplified according to the case, a qualitative study of the system of equations is presented and conclusions are drawn regarding the future situation in relation to the number of infested patients.

This work is funded by FEDER funds through the COMPETE 2020 Program, National Funds through FCT - Portuguese Foundation for Science and Technology and POR Lisboa2020 , under the projects with references POCI- 01-0145-FEDER-007688 (Reference UID/CTM/50025), UID/BIA/00329/2013, PTDC/CTM-BIO/6178/2014, M-ERA-NET2/0007/2016 (CellColor) and PTDC/CTM-REF/30529/2017. S.N. Fernandes and L.F. Lopes acknowledge the Minister of Science, Technology and Higher Education for National Funds , European Social Funds and FCT for fellowships number SFRH/BDP/78430/2011 and SFRH/BPD/84478/2012 , respectively. This work was supported by the Histology and Comparative Pathology laboratory [Electron Microscopy service] of the Instituto de Medicina Molecular João Lobo Antunes, specifically Andreia Pinto for her kind support. We would like to thank Carlos Silva for providing the C. chrysina specimen. The C. aurata specimen observed was provided by the Insect Collection of the National Museum of Natural History and Science, University of Lisbo Significant advances have been made to control the iridescence and the selective reflection of left circularly polarised (LCP) light, and transmission of right circularly polarised (RCP) light of solid films prepared from cellulose nanocrystals (CNCs). However the manipulation of the photonic properties of the CNCs films, which reflect both RCP and LCP light is less investigated. Solid films prepare from natural sources as CNCs have advantageous characteristics that are absent in other synthetic structures, such as wide availability and renewability. Here we review and compare recent research activity involving the production and characterization of photonic band gap structures resulting from an anisotropic layer inserted between two cholesteric layers with different helical pitches but the same handedness. We make connections between systems existing in Nature and synthetic ones with the hope of advancing in the production and manipulation of CNCs-based photonic structures. authorsversion published

The colloidal suspensions of aqueous cellulose nanocrystals (CNCs) are known to form liquid crystalline (LC) systems above certain critical concentrations. From an isotropic phase, tactoid formation, growth, and sedimentation have been determined as the genesis of a high-density cholesteric phase, which, after drying, originates solid iridescent films. Herein, the coexistence of a liquid crystal upper phase and an isotropic bottom phase in CNC aqueous suspensions at the isotropic-nematic phase separation is reported. Furthermore, isotropic spindle-like domains are observed in the low-density LC phase and high-density LC phases are also prepared. The CNCs isolated from the low- and high-density LC phases are found to have similar average lengths, diameters, and surface charges. The existence of an LC low-density phase is explained by the presence of air dissolved in the water present within the CNCs. The air dissolves out when the water solidifies into ice and remains within the CNCs. The self-adjustment of the cellulose chain conformation enables the entrapment of air within the CNCs and CNC buoyancy in aqueous suspensions.

Funding: We wish to thank all the collaborators (administrative staff, nurses, etc.) of the surgery block, as well as the doctors of the vascular surgery and transplantation departments of the Hospital of Santa Maria for the availability and assistance in the collection of the samples. We also thank Sociedade Portuguesa de Reumatologia for funding with two fellowships: Fundo de Apoio à Investigação 2014 and SPR/MSD 2015. DC-F received funding from a PhD grant from Fundação para a Ciência e a Tecnologia (SFRH/BD/80940/2011). Background and Aims: Atherosclerosis and osteoporosis share common risk factors, as well as inflammatory mechanisms. Our aim was to understand how atherosclerotic lesions are related with disturbances in bone. Methods: Gene expression of pro-inflammatory and bone metabolism related proteins (IL-1β, IL-6, IL-17A, TNF, RANKL, OPG, COL1, CTSK, OCL, TRAP, CBFA1, DKK1, SOST, ADIPOQ, and ADIPOR1) were analyzed in arteries and bones from 45 deceased donors and adipose tissue was used as control. Additionally, in 139 patients with advanced atherosclerosis submitted to carotid endarterectomy we compared calcium content (Alizarin red) and plaque inflammatory scores (CD3+, CD68+, and adiponectin) of patients with normal bone mineral density (BMD) with those with low BMD and explored the associations between gene expression in atherosclerotic plaques and BMD. Serum levels of pro-inflammatory and bone related proteins were measured both in donors and patients. Associations were investigated by the Pearson or Spearman correlation tests, and multivariate regression analyzes were performed when justified. Results: Gene expression of bone remodeling and pro-inflammatory proteins correlated positively in bone and aorta, independently of age and sex of donors, but not in adipose tissue. The expression of bone formation genes was significantly higher in atheroma plaques from endarterectomized patients with normal vs. low BMD as well as inflammatory CD68+ scores, regardless of patients' age and sex, but not of body mass index. No relationship was observed between serum levels and gene expression levels of pro-inflammatory or bone remodeling proteins. Conclusions: Our results suggest that the relationship between bones and vessels in the context of atherosclerotic disease and osteoporosis may rely on the intrinsic connection between the tissues involved, independently of disease stage. Serum measurements of pro-inflammatory and bone-remodeling proteins do not accurately translate tissue pathologic processes. publishersversion published

Winning the war against resistant bacteria will require a change of paradigm in antibiotic discovery. A promising new direction is the targeting of non-essential pathways required for successful infection, such as quorum-sensing, virulence, and biofilm formation. Similarly important will be strategies to prevent or revert antibiotic resistance. Here, we argue that the (p)ppGpp signaling pathway should be a prime target of this effort, since its inactivation could potentially achieve all these goals simultaneously. The hyperphosphorylated guanine nucleotide (p)ppGpp is an ancient and universal second messenger of bacteria that has pleotropic effects on the physiology of these organisms. Initially described as a stress signal—an alarmone—it is now clear that (p)ppGpp plays a more general and fundamental role in bacterial adaptation, by integrating multiple internal and environmental signals to establish the optimal balance between growth and maintenance functions at any given time. Given such centrality, perturbation of the (p)ppGpp pathway will affect bacteria in multiple ways, from the ability to adjust metabolism to the available nutrients to the capacity to differentiate into developmental forms adapted to colonize different niches. Here, we provide an overview of the (p)ppGpp pathway, how it affects bacterial growth, survival and virulence, and its connection with antibiotic tolerance and persistence. We will emphasize the dysfunctions of cells living without (p)ppGpp and finalize by reviewing the efforts and prospects of developing inhibitors of this pathway, and how these could be employed to improve current antibiotic therapy.

It costs money to improve the welfare of farm animals. For people with animals under their care, there are many factors to consider regarding changes in practice to improve welfare, and the optimal course of action is not always obvious. Decision support systems for animal welfare, such as economic cost–benefit analyses, are lacking. This review attempts to provide clarity around the costs and benefits of improving farm animal welfare, thereby enabling the people with animals under their care to make informed decisions. Many of the costs are obvious. For example, training of stockpeople, reconfiguration of pens, and administration of pain relief can improve welfare, and all incur costs. Other costs are less obvious. For instance, there may be substantial risks to market protection, consumer acceptance, and social licence to farm associated with not ensuring good animal welfare. The benefits of improving farm animal welfare are also difficult to evaluate from a purely economic perspective. Although it is widely recognised that animals with poor welfare are unlikely to produce at optimal levels, there may be benefits of improving animal welfare that extend beyond production gains. These include benefits to the animal, positive effects on the workforce, competitive advantage for businesses, mitigation of risk, and positive social consequences. We summarise these considerations into a decision tool that can assist people with farm animals under their care, and we highlight the need for further empirical evidence to improve decision-making in animal welfare.

This article belongs to the Special Issue Cytotoxicity and Genotoxicity Assessment in Nanomaterials. Cellulose micro/nanomaterials (CMNM), comprising cellulose microfibrils (CMF), nanofibrils (CNF), and nanocrystals (CNC), are being recognized as promising bio-nanomaterials due to their natural and renewable source, attractive properties, and potential for applications with industrial and economical value. Thus, it is crucial to investigate their potential toxicity before starting their production at a larger scale. The present study aimed at evaluating the cell internalization and in vitro cytotoxicity and genotoxicity of CMNM as compared to two multi-walled carbon nanotubes (MWCNT), NM-401 and NM-402, in A549 cells. The exposure to all studied NM, with the exception of CNC, resulted in evident cellular uptake, as analyzed by transmission electron microscopy. However, none of the CMNM induced cytotoxic effects, in contrast to the cytotoxicity observed for the MWCNT. Furthermore, no genotoxicity was observed for CNF, CNC, and NM-402 (cytokinesis-block micronucleus assay), while CMF and NM-401 were able to significantly raise micronucleus frequency. Only NM-402 was able to induce ROS formation, although it did not induce micronuclei. Thus, it is unlikely that the observed CMF and NM-401 genotoxicity is mediated by oxidative DNA damage. More studies targeting other genotoxicity endpoints and cellular and molecular events are underway to allow for a more comprehensive safety assessment of these nanocelluloses. This research received funding from the Portuguese Foundation for Science and Technology (FCT/MCTES), through national funds (PIDDAC) under the project ToxApp4NanoCELFI (PTDC/SAUPUB/32587/2017). The following financial grants are also acknowledged: Projects UIDB/00009/2020 and UIDP/00009/2020 (ToxOmics); Projects UIDB/04138/2020 and UIDP/04138/2020 (iMed.Ulisboa) and L. Gonçalves Principal Researcher grant (CEECIND/03143/2017); FEDER and POR Lisboa2020 through the COMPETE 2020 POCI and PORL; POCI-01-0145-FEDER-007688 (UIDB/50025/2020- 2023); NanoCell2SEC (PTDC/CTM-REF/30529/2017); CIEPQPF (UIDB/00102/2020); European Topology Interdisciplinary Action (EUTOPIA CA17139); Inpactus Project—Innovative Products and Technologies from Eucalyptus (Project Nº 21874, funded by Portugal 2020 through ERDF in the frame of COMPETE 2020 Nº246/AXIS II/2017). Nádia Vital holds an FCT/MCTES PhD Scholarship grant (2020.07168.BD) info:eu-repo/semantics/publishedVersion

Snap and common beans (Phaseolus vulgaris L.) are severely affected by Bean golden mosaic virus (BGMV) infection, so far the only begomovirus reported on these crops in Brazil (1). Samples of snap and common beans colonized by the whitefly Bemisia tabaci biotype B and displaying golden mosaic, chlorotic spots, and leaf distortion were collected in three production regions in Goiás State (Goianápolis, Luziânia, and Itaberaí) between 2003 and 2007. Total DNA extracted from leaf samples was used as template in PCR assays using universal primers targeting conserved regions of the DNA-A and DNA-B genomes (3). Begomovirus-specific amplicons were observed only with DNA template from symptomatic plants. Two single amplicons were observed for both genomic segments, indicating the presence of bipartite species in all samples. Sequence analysis of four isolates (named as GO-176, GO-260, GO-354, and GO-368) obtained from common bean samples indicated identity levels of approximately 95% with the DNA-A segment of BGMV (GenBank Accession No. FJ665283). However, the complete DNA-A sequence (GenBank Accession No. HM357459.1) of the GO-060 isolate (from a symptomatic snap bean plant collected in Goianápolis) displayed 76% identity with BGMV (GenBank Accession No. FJ665283) and 95% identity with the DNA-A of a Sida micrantha mosaic virus (SimMV) isolate (GenBank Accession No. EU908733.1) reported to be infecting okra (Abelmoschus esculentus L.) and 94.8% with a SimMV isolate reported to be infecting soybean (GenBank Accession No. FJ686693) in Brazil (2). Koch's postulates were fulfilled for the isolate GO-060 by inoculating a set of soybean and bean accessions via a biolistic approach. The ratio of positive PCR amplicons per total of inoculated plants were 15 of 16 for snap bean cv. Trepador, 9 of 10 for snap bean cv. Fartura, 18 of 24 for common bean cv. Olate Pinto, and 19 of 25 for common bean cv. Carioca. The isolate was also able to infect eight of nine soybean ‘Doko’ plants. Sequence analysis using symptomatic leaf samples (15 days after inoculation) confirmed SimMV as the causal agent. To our knowledge, this is the first report of a SimMV isolate infecting P. vulgaris. This virus is apparently fast expanding its host range from Malvaceae to Solanaceae species and leguminous hosts after the introduction of B. tabaci biotype B (2). More extensive surveys are necessary to access the current epidemiological importance of SimMV in both snap and common beans in Brazil. References: (1) J.C. Faria and D. P. Maxwell. Phytopathology 89:262, 1999. (2) F. R. Fernandes et al. Arch. Virol. 154:1567, 2009. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993.

Cellulose nano crystals (CNCs) are promising materials for energy efficient buildings related to the control of reflectivity and heat absorption/reflection of light. In this sense it is important to improve CNCs films fire retardant properties, which can be achieved by adding clays. Cellulose nanocrystals (CNCs) and nanolayers obtained from Sodium Fluorohectorite (NaFh) synthetic clay are both known to form liquid crystalline phases in aqueous suspensions. CNCs form cholesteric phases, which structure is preserved after water evaporation, while dry NaFh nanolayers aligned films collapse. In this initial work, it is shown that CNCs are compatible with NaFh clay. We demonstrate that the liquid crystalline phase of CNCs in water is not destroyed by the presence of NaFh nanolayers. The NaFh nanolayers act as planar anchoring surfaces to the cellulose nanorods and, after evaporation of the water coloured films are obtained. The precursor solutions and the photonic films were investigated by Describe several techniques. Open Access This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

BackgroundExisting diagnostic methods for the parasitic gastrointestinal nematode,Haemonchus contortus, are time consuming and require specialised expertise, limiting their utility in the field. A practical, on-farm diagnostic tool could facilitate timely treatment decisions, thereby preventing losses in production and flock welfare. We previously demonstrated the ability of visible–near-infrared (Vis–NIR) spectroscopy to detect and quantify blood in sheep faeces with high accuracy. Here we report our investigation of whether variation in sheep type and environment affect the prediction accuracy of Vis–NIR spectroscopy in quantifying blood in faeces.MethodsVisible–NIR spectra were obtained from worm-free sheep faeces collected from different environments and sheep types in South Australia (SA) and New South Wales, Australia and spiked with various sheep blood concentrations. Spectra were analysed using principal component analysis (PCA), and calibration models were built around the haemoglobin (Hb) wavelength region (387–609 nm) using partial least squares regression. Models were used to predict Hb concentrations in spiked faeces from SA and naturally infected sheep faeces from Queensland (QLD). Samples from QLD were quantified using Hemastix® test strip and FAMACHA© diagnostic test scores.ResultsPrincipal component analysis showed that location, class of sheep and pooled versus individual samples were factors affecting the Hb predictions. The models successfully differentiated ‘healthy’ SA samples from those requiring anthelmintic treatment with moderate to good prediction accuracy (sensitivity 57–94%, specificity 44–79%). The models were not predictive for blood in the naturally infected QLD samples, which may be due in part to variability of faecal background and blood chemistry between samples, or the difference in validation methods used for blood quantification. PCA of the QLD samples, however, identified a difference between samples containing high and low quantities of blood.ConclusionThis study demonstrates the potential of Vis–NIR spectroscopy for estimating blood concentration in faeces from various types of sheep and environmental backgrounds. However, the calibration models developed here did not capture sufficient environmental variation to accurately predict Hb in faeces collected from environments different to those used in the calibration model. Consequently, it will be necessary to establish models that incorporate samples that are more representative of areas whereH. contortusis endemic.Graphical Abstract

Background: Existing diagnostic methods for the parasitic gastrointestinal nematode, Haemonchus contortus, are time consuming and require specialised expertise, limiting their utility in the field. A practical, on-farm diagnostic tool could facilitate timely treatment decisions, preventing production and welfare loss in the flock. We previously demonstrated the ability of visible-near infrared (vis-NIR) spectroscopy to detect and quantify blood in sheep faeces with high accuracy. Here we investigate whether variation in sheep type and environment affect the prediction accuracy of vis-NIR spectroscopy in quantifying blood in faeces.Methods: Vis-NIR spectra were obtained from worm-free sheep faeces from different environments in South Australia (SA) and New South Wales (NSW), Australia and spiked with various sheep blood concentrations collected. Spectra were analysed using principal component analysis (PCA), and calibration models were built around the haemoglobin (Hb) wavelength region (387 – 609 nm) using partial least squares (PLS) regression. Models were used to predict Hb concentrations in spiked faeces from SA and naturally infected Queensland (QLD) faeces. Naturally occurring blood in QLD samples was quantified using Hemastix® and FAMACHA© scores.Results: PCA showed that location, class of sheep and pooled/individual samples were factors affecting the Hb predictions in sheep faeces. The calibration models successfully differentiated ‘healthy’ SA samples from those requiring anthelmintic treatment with moderate to good prediction accuracy (sensitivity: 57 – 94%, specificity: 44 – 79%). The models were not predictive for naturally infected QLD samples, which may be due in part to variability of faecal background and blood chemistry between samples, or the difference in validation methods used for blood quantification. PCA of QLD samples, however, identified a difference between samples containing high and low quantities of blood.Conclusion: This study demonstrates the potential of vis-NIR spectroscopy for estimating blood concentration in faeces from various types of sheep and environmental backgrounds. However, the calibration models developed here did not capture enough environmental variation to accurately predict Hb in faeces collected from environments different to those used in the calibration model. Consequently, it will be necessary to establish models that incorporate samples that are more representative of areas where H. contortus is endemic for the accurate prediction of H. contortus infections in these regions.

POCI- 01-0145-FEDER-007688 (Reference UIDB/50025/2020-2023) PTDC/CTM-BIO/6178/2014 M-ERA-NET2/0007/2016 PTDC/CTM-REF/30529/2017 EUTOPIA CA17139 Slovenian Research Agency Grant Z1-5441 P1-0099 Structural vivid colours can arise from the interference of light reflected from structures exhibiting periodicity on scales in the range of visible wavelengths. This effect is observed with light reflected from cell-walls of some plants and exoskeletons of certain insects. Sometimes the colour sequence observed for these structures consists of nearly circular concentric rings that vary in colour from Red, Orange, Yellow, Green, Cyan to Blue, from the periphery to the centre, similarly to the colour scheme sequence observed for the rainbow (ROYGB). The sequence of colours has been found for solid films obtained from droplets of aqueous cellulose nanocrystals (CNCs) suspensions and attributed to a "coffee ring"effect. In this work, coloured lyotropic solutions and solid films obtained from a cellulose derivative in the presence of trifluoroacetic acid (TFA), which acts as a "reactive solvent", are revisited. The systems were investigated with spectroscopy, using circularly and linearly polarised light, coupled with a polarised optical microscope (POM) and scanning electron microscopy (SEM). The lyotropic cholesteric liquid crystalline solutions were confined in capillaries to simplify 1D molecular diffusion along the capillary where an unexpected sequence of the structural colours was observed. The development and reappearance of the sequence of vivid colours seem consistent with the reaction-diffusion of the "reactive solvent"in the presence of the cellulosic chains. The strong TFA acts as an auto-catalyst for the chemical reaction between TFA and the hydroxyl groups, existing along the cellulosic chain, and diffuses to the top and bottom along the capillaries, carrying dissolved cellulosic chains. Uncovering the precise mechanism of colour sequence and evolution over time in cellulosic lyotropic solutions has important implications for future optical/sensors applications and for the understanding of the development of cellulose-based structures in nature. This journal is authorsversion published

Wolbachia are maternally transmitted intracellular bacteria that naturally infect over 40% of all insect species. Wolbachia have become promising biocontrol agents against mosquito-borne diseases due to their unique effects on mosquito reproduction and immunity. However, the phenotypes mediated by Wolbachia vary between strains and host-pathogen systems. Discovery of novel Wolbachia strains is essential for broadening the application of Wolbachia for mosquito control. In this study, I identified and characterized the anti-pathogen effects of natural Wolbachia infections in Australian mosquitoes. I identified three previously uncharacterized Wolbachia strains based on detection and sequencing of wsp, 16S rRNA and ftsZ genes. Wolbachia sequences were also detected from Coquillettidia xanthogaster, however, it remained unclear if the detection was resulted from a genuine infection. I established colonies of two local species that are naturally infected with Wolbachia; Aedes notoscriptus and Culex sitiens, and discovered a unique pattern of infection rates fluctuating between 15% and 60% in both colonies. Study on the maternal transmission of Wolbachia in Cx. sitiens revealed high transmission rates (99.7%) but low cytoplasmic incompatibility (9.9% mortality). I orally inoculated Wolbachia positive and negative Ae. notoscriptus and Cx. sitiens with the alphavirus Ross River virus (RRV; 103.31 – 6.48 and 105.41 – 6.36 CCID50 per mosquito, respectively). RRV infection rates varied between 17% - 85% for Ae. notoscriptus and 0% - 18% for Cx. sitiens with no significant differences between infection rates for Wol+ and Wol- mosquitoes. However, mean virus load in Wol+ Cx. sitiens was 1000× lower than in Wol- mosquitoes, suggesting that natural Wolbachia infection suppresses virus proliferation in that species. I report on progress towards transinfecting cells (Aag2 and Aa20) and mosquitoes (Aedes aegypti) with Wolbachia, including oral ingestion trials. Inoculations using the Shell vial technique resulted in temporal in vitro infections for up to three passages, however, stable infections were not yet achieved. I observed evidence that Wolbachia infected cells within the epithelial lining of the crop and potentially the midgut by fluorescence microscopy, however, the infection was transient. These discoveries broadened the current understanding on Wolbachia-host-pathogen interactions and shed light on future mosquito control strategies. The behaviour of Wolbachia in new hosts can be difficult to predict, therefore transinfection and further characterization is required to determine if these strains can induce anti-pathogen effects in the context of mosquito control.

Resumo Fundamento A prevalência da obesidade vem aumentando sistematicamente na população, inclusive nas crianças e adolescentes, ao redor do mundo. Objetivos Descrever curvas percentílicas de referência para a circunferência abdominal (CA) nas crianças brasileiras e fornecer pontos de corte da CA para identificar crianças com risco de obesidade. Métodos Um estudo multicêntrico, prospectivo, tranversal foi realizado em crianças com idades entre 6 e 10 anos, matriculadas no ensino fundamental de escolas públicas e particulares de 13 cidades do estado de São Paulo. A estatura, o peso e a CA foram medidos em duplicata em 22.000 crianças (11.199 meninos). Para estabelecer o melhor ponto de corte da CA para o diagnóstico da obesidade, foram calculadas curvas ROC com crianças classificadas como com peso normal e obesas, de acordo com as curvas do IMC, estratificadas por gênero e idade, e o índice Youden foi utilizado como a eficácia potencial máxima desse biomarcador. Valores de p < 0,05 foram considerados estatisticamente significativos. Resultados Os valores da CA aumentaram com a idade, tanto nos meninos quanto nas meninas. A prevalência de obesidade em cada grupo variou de 17% (6 anos de idade) a 21,6 % (9 anos de idade), dentre os meninos, e de 14,1% (7 anos de idade) a 17,3 % (9 anos de idade), dentre as meninas. As análises ROC mostraram o percentil 75 como ponto de corte para o risco de obesidade, e o diagnóstico de obesidade está classificado no percentil 85 ou acima. Conclusão Curvas de referência da CA específicas para idade e sexo em crianças brasileiras e pontos de corte para o risco de obesidade podem ser usados em triagem nacional e estudos intervencionais para reduzir a carga da obesidade no Brasil. (Arq Bras Cardiol. 2020; 114(3):530-537) Abstract Backgroud The prevalence of obesity has systematically been increased in the population, including children and adolescents, around the world. Objectives To describe reference percentile curves for waist circumference (WC) in Brazilian children and provide cut-off values of WC to identify children at risk for obesity. Methods A multicenter, prospective, cross-sectional study was performed with children aged from 6 to 10 years old, enrolled in public and private elementary schools from 13 cities of the São Paulo State. Height, weight, and WC were measured in duplicate in 22,000 children (11,199 boys). To establish the WC best cut-off value for obesity diagnosis, ROC curves with children classified as normal weight and obese were calculated, according to BMI curves, stratified by gender and age, and the Youden Index was utilized as the maximum potential effectiveness of this biomarker. A p < 0.05 was considered statistically significant. Results WC values increased with age in both boys and girls. The prevalence of obesity in each age group varied from 17% (6 years old) to 21.6% (9 years old) among boys, and from 14.1% (7 years old) to 17.3 % (9 years old) among girls. ROC analyses have shown the 75th percentile as a cut-off for obesity risk, and the diagnosis of obesity is classified on the 85th percentile or more. Conclusion Age and gender specific reference curves of WC for Brazilian children and cut-off values for obesity risk may be used for national screening and interventional studies to reduce the obesity burden in Brazil. (Arq Bras Cardiol. 2020; 114(3):530-537)

Heavy infestations of the Barber’s pole worm, Haemonchus contortus, can cause severe wasting, morbidity and mortality in animals if not promptly treated. The current detection methods for this blood-sucking parasite involve faecal worm egg counts and diagnosis of anaemia, both of which are time consuming and require expertise. As blood is detected in sheep faeces sooner during infection than worm eggs, quantitative evaluation of blood may serve as a sensitive indicator of H. contortus infection. Here we investigated the feasibility of rapid estimation of haemoglobin (Hb) in sheep faeces using visible near-infrared spectroscopy. Haemoglobin (Hb) was assessed at various concentrations in moist sheep faeces using portable visible near-infrared (vis-NIR) spectrometers. Calibration models were developed for the region of 400–600 nm, where Hb absorption bands can be found. Within this wavelength region, Hb in sheep faeces can be estimated with minimal interference from background moisture (970 nm) or chlorophyll (670 nm), suggesting that difference in diets in sheep will have minimal effect on prediction accuracy. Predictions for sheep faeces based on a drenching threshold of 3 µg Hb/mg faeces showed high levels of accuracy with minimal sample preparation (Sensitivity = 89%, specificity = 80%). The success in detecting Hb in sheep faeces indicates the potential of vis-NIR spectroscopy as a rapid, on-farm diagnostic method for predicting blood in sheep faeces, and timing treatment of H. contortus infections.

FCT – Portuguese Foundation for Science and Technology through the Ph.D. scholarship SFRH/BD/125191/2016. project PapEl, reference PTDC/CTM-NAN/5172/2014. project CHIHC, reference PTDC/NAN-MAT/32558/2017. project PTDC/CTM-BIO/6178/2014, M-ERA-NET2/0007/2016 (CellColor). POR Lisboa2020 through project PTDC/CTM-REF/30529/2017. D.G. acknowledges the support from FCT – Portuguese Foundation for Science and Technology through the AdvaMTech PhD program scholarship PD/BD/52627/2014. Mesoporous structures made of cellulose nanocrystals (CNCs) and their self-assembly into films are of great interest not only due to their abundancy and sustainability but also due to their ease of chemical modification and nanoscale biomimicry capabilities. However, their implementation in (opto)electronic devices requires further understanding on how these self-assembled twisted mesoporous superstructures respond to electrical stimulus. In this regard, this work focuses on the infiltration of solid CNC droplets with three distinct alkali ions (Li+, Na+, and K+) to yield films with improved electrochemical response when compared to pristine ones, while preserving their photonic character. Electrochemical characterization shows capacitances of up to 2.5 μF cm–2 allowing for their integration as solid-state gate electrolytes in amorphous indium–gallium–zinc–oxide transistors, resulting in low operating voltages (10 cm2 V–1 s–1. Devices fabricated on Na+ and K+ infiltrated CNC films present the best characteristics, indicating pure capacitive charging of the semiconductor. The insights presented here contribute to applications in solid-state ionics in mesoporous structures or the combination of optically active electrolytes capable of providing unique functionalities in ion-gated transistors and circuitry. publishersversion published

FEDER funds through the COMPETE 2020 Program, National Funds through FCT -Portuguese Foundation for Science and Technology and POR Lisboa2020, PTDC/CTM-BIO/6178/2014, M-ERA-NET2/0007/2016 (CellColor) and PTDC/CTM-REF/30529/2017 (NanoCell2SEC). Lyotropic colloidal aqueous suspensions of cellulose nanocrystals (CNCs) can, after solvent evaporation, retain their chiral nematic arrangement. As water is removed the pitch value of the suspension decreases and structural colour-generating films, which are mechanically brittle in nature, can be obtained. Increasing their flexibility while keeping the chiral nematic structure and biocompatible nature is a challenging task. However, if achievable, this will promote their use in new and interesting applications. In this study, we report on the addition of different amounts of hydroxypropyl cellulose (HPC) to CNCs suspension within the coexistence of the isotropic-anisotropic phases and infer the influence of this cellulosic derivative on the properties of the obtained solid films. It was possible to add 50 wt.% of HPC to a CNCs aqueous suspension (to obtain a 50/50 solids ratio) without disrupting the LC phase of CNCs and maintaining a left-handed helical structure in the obtained films. When 30 wt.% of HPC was added to the suspension of CNCs, a strong colouration in the film was still observed. This colour shifts to the near-infrared region as the HPC content in the colloidal suspension increases to 40 wt.% or 50 wt.% The all-cellulosic composite films present an increase in the maximum strain as the concentration of HPC increases, as shown by the bending experiments and an improvement in their thermal properties. publishersversion published