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The Covid-19 pandemic turned educational institutions from face-to-face to modular learning modality. Through the implementation of distance learning scheme in the Philippines, Eastern Samar State University (ESSU) also adapted this new normal learning strategy through development of mobile-based course modules for distance learning as an alternative to its present modular distance learning approach. Paper-based modules seemed to be expensive and sometimes texts are not clear and readable. Thus, students get bored and perform low at school. As a response, a mobile-based course learning module is developed for offline learners. This study employs Waterfall model in the development of the application. This model follows several phases from designing, developing and testing the application to ensure its quality. The findings of the study show that the application is highly usable and functional after series of tests conducted. Moreover, it gained 88.75 overall score in the System Usability test conducted. This implies that the application is usable, user-friendly, functional, and portable. A further study may be conducted on the effectiveness of the mobile application in the instruction.

BACKGROUND: The strawberry is a fruit with great acceptance by the consumer due to its pleasant flavor and functionality; It is rich in vitamin C, tannins, flavonoids, anthocyanins, catechin, quercetin and kaempferol, organic acids and minerals. OBJETIVE: The objective of the research was to evaluate the effect of the feed formulation and the spray-drying process on the quality of strawberry extract microcapsules (EMs). METHODS: The response surface methodology was used with a central composite design centered on the face (α= 1), considering the factors: maltodextrin (MD), inlet air temperature (IAT), outlet air temperature (OAT) and atomizing disk speed (ADS), and the response variables: humidity (Xw), water activity (aw), solubility (S), wettability (We), hygroscopicity (Hy), bulk density (ρb), compacted density (ρc), total phenols (TP), antioxidant capacity, angle of repose (AR), strawberry powder particle size (D[3 ;2]), color (CIE-Lab), and process yield (Y). RESULTS: The aw and Xw values guarantee microbiological stability and control over degradation reactions in EMs. For the levels evaluated, the S was affected by OAT and ADS, and interaction IAT*MD, ADS*MD, IAT2, and MD2. The We depended on the factors IAT, OAT, and the interactions IAT*OAT and IAT*MD. ABTS activity was affected by the factors ADS, ADS*MD and IAT2. The Y was affected by OAT, MD, IAT*OAT, IOT*ADS, IOT*MD, OAT*ADS, ADS*MD, IOT2, and OAT2. The increase in MD caused a decrease in Hy. The value of L* was affected by ADS and MD, effects due to temperature did not affect the levels evaluated, and chromaticity a* was positively affected by the increase in ADS. IAT and MD presented a statistical effect on AR. CONCLUSIONS: The experimental optimization reached a desirability of 73.7%, being the optimal conditions: IAT (154°C), OAT (89°C), ADS (16,805 rpm), and MD (11.5%).

BACKGROUND: There is consumer interest in berries due to their nutritional properties, especially biologically active compounds. Spray drying technologies improve products stability and shelf life, and the agglomeration process subsequently enhances powder instantaneousness and flow properties. OBJECTIVE: The aim was to evaluate the effect of the fluidized bed agglomeration process on flow, reconstitution, and antioxidant properties of cape gooseberry, strawberry, and blackberry powder mix obtained by spray drying and a water and vitamin C binding solution. METHODS: A face-centered central composite design with 15 experiments was used; independent variables were fluidization temperature (T), binder solution atomization pressure (P), and process time (t), while dependent variables were moisture, solubility, wettability, particle size, antioxidant capacity, total phenols, Vit. C, β-carotenes (β-car), and total anthocyanins. RESULTS: The best process conditions from experimental optimization were T = 70 °C, P = 1.1 bar, and t = 29.1 min. This agglomerated product had low moisture values, good solubility, and 0.5 min wettability; the values for Vit. C = 246.4±3.4 mg/100 g db, β-car = 4.0±0.5 mg/100 g db, and total anthocyanins = 230.7±3.0 mg C3G/100 g db were noteworthy. CONCLUSIONS: An agglomerate berry-based powder product was obtained with 100% Vit. C RDA (Recommended Dietary Allowances) and 15% vitamin A RDA. Flow and instantaneousness properties of the agglomerated powder also significantly improved with the applied process.

The efficacy of a vector control tool in reducing mosquito biting is crucial for its acceptability. The present study compared the vector density of Culex spp. And Mansonia spp. across clusters, which received two dual-active ingredient (a.i.) long-lasting insecticidal nets (LLINs) and a standard pyrethroid-only LLIN, and assessed the seasonality of these mosquito genera. A total of 85,723 Culex spp. and 144,025 Mansonia spp. were caught over the study period. The density of Culex and Mansonia was reduced in all three arms over the study period. There was no evidence of a significant reduction in the indoor or outdoor density of Culex spp. in either dual-a.i. LLIN arm as compared to the standard pyrethroid-only net arm. A similar trend was observed with Mansonia spp. A high density of Culex spp. was found both in rainy and dry seasons, while for Mansonia spp., this was mainly observed during the rainy season. These results suggest that the novel insecticides in the dual-a.i. LLINs did not have an additional impact on these species and that pyrethroids might still be effective on them. Further work is required to determine whether these species of mosquitoes have resistance to the insecticides tested in this trial.

Additive Manufacturing is an emerging technology used in wide applications including, wastewater treatment. This paper presents applied techniques of Additive Manufacturing in producing a 3D-printed prototype model of a laboratory-scale wastewater treatment process. This paper aims to show the efficiency of bioremediation of freshwater algae in reducing traces of phosphate and nitrates - both known pollutants responsible for eutrophication. The study also comparatively reviews two different set-ups: stagnant and aerated. 3D printed bio-propeller, a polylactic acid-based aerator, was applied to the latter aforementioned set-up and was designed to be durable, environmental oriented, and cost-efficient. Moreover, the features and details of the 3D-printed bio-propeller emphasized its ability to promote algae harvesting by acting as a sustainable biofilm for algal growth. It is found that both set-ups show significant algae bioremediation efficiency for the synthetic nutrients. Also, the aerated set-up indicated a favorable result with the highest efficiency of 92.857% for nitrates and 41.667% for phosphate. It is concluded that the potentials of 3D printed bio-propeller as a prime component of Rotating Biological Contractor (RBC) can promote sustainable wastewater treatment.

Mobile Technology plays significant contributions to the humanity in many areas. One of these is on the issues on Disaster Risk Reduction Management through developments of mobile applications. This study primarily aims to design, develop and test the Hazard Safety Tips Mobile Application in Waray-waray dialect. This study is conducted to help the residents of Salcedo, Eastern Samar become more aware of how they will be able to manage their safety in times of calamities and disaster through the aid of this application. Waterfall model is used as System Development Life Cycle (SDLC) which includes the following phases: Requirement Definition, System and Software Design, Implementation and Unit Testing, Integration and System Testing and Operation and Maintenance. The researchers gathered data in the Municipal Disaster Risk Management Office and its website which served as inputs in the development of the application. Flowcharts and data flow diagrams were created for the designing of the application. User interfaces were designed and developed using Android Studio. The application had undergone three testing strategies: Benchmark, Alpha and Beta Tests using ISO scorecard. Results showed that in terms of usability, the application scored 88.75, which is above average. This means that the developed application was a great help in terms of hazard and disaster preparedness to the residents of Salcedo, Eastern Samar and the region as a whole. The study concluded that the mobile application was effective and functional because it conforms to the specified requirements

Chemical communication is an important mechanism of mate choice across the animal tree of life. However, anthropogenic perturbation of the signaling environment can disrupt chemical communication and result in a breakdown of behavioral reproductive isolation. Here we find that calcium hydroxide (Ca(OH)2), a common and deliberately introduced anthropogenic pollutant, profoundly disrupts chemical communication in the swordtail fish Xiphophorus birchmanni. Moreover, it acts in a way that should promote hybridization, causing female X. birchmanni to prefer the chemical cues of the parapatric sister species X. malinche. We find that this flip in the direction of preference is attributable both to a reduced preference for conspecific signals and to a coupled strengthening of preference for sister-species signals.

Adaptive plasticity requires an integrated suite of functional responses to environmental variation, which can include social communication across life stages. Desert locusts (Schistocerca gregaria) exhibit an extreme example of phenotypic plasticity called phase polyphenism, in which a suite of behavioral and morphological traits differ according to local population density. Male and female juveniles developing at low population densities exhibit green- or sand-colored background-matching camouflage, while at high densities they show contrasting yellow and black aposematic patterning that deters predators. The predominant background colors of these phenotypes (green/sand/yellow) all depend on expression of the carotenoid-binding "Yellow Protein" (YP). Gregarious (high-density) adults of both sexes are initially pinkish, before a YP-mediated yellowing reoccurs upon sexual maturation. Yellow color is especially prominent in gregarious males, but the reason for this difference has been unknown since phase polyphenism was first described in 1921. Here, we use RNA interference to show that gregarious male yellowing acts as an intrasexual warning signal, which forms a multimodal signal with the antiaphrodisiac pheromone phenylacetonitrile (PAN) to prevent mistaken sexual harassment from other males during scramble mating in a swarm. Socially mediated reexpression of YP thus adaptively repurposes a juvenile signal that deters predators into an adult signal that deters undesirable mates. These findings reveal a previously underappreciated sexual dimension to locust phase polyphenism, and promote locusts as a model for investigating the relative contributions of natural versus sexual selection in the evolution of phenotypic plasticity. ispartof: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA vol:119 issue:34 ispartof: location:United States status: published

Resumen: Introducción: El fútbol infantil y juvenil es un fenómeno cultural y social de gran importancia en Argentina. Objetivo: Este trabajo tiene como objetivo dar cuenta de aspectos fundamentales de la estructura organizativa del fútbol infantil y juvenil en la liga de Mar del Plata, una de las ciudades más importantes del país. Métodos: El análisis de datos cuantitativos básicos permite dar cuenta de los aspectos fundamentales de la estructura organizativa del fútbol infantil y juvenil en esta liga. Este análisis elemental de cuantitativo se combina con una interpretación de informes basada tanto en un análisis discursivo como etnográfico. Resultados y discusión: Existe un severo proceso de exclusión y abandono en la liga de fútbol de Mar del Plata (LMF). No sólo ocurre que el número de jugadoras mujeres es extremadamente bajo en comparación con el número de jugadores varones, sino que también ocurre que más de mil varones son abandonados año tras año por la LMF. El complejo proceso de abandono y exclusión también está fuertemente ligado a una serie de fenómenos diversos, como el sexismo, la ausencia de personas con discapacidad, la imitación de prácticas de fútbol profesional y el uso de etiquetas y estereotipos. Conclusiones: Dado que promueve sistemáticamente la exclusión y el abandono, el LMF no puede cumplir una función educativa o inclusiva, y ni siquiera puede ayudar a formar jugadores profesionales de alto nivel. En su estado actual, la estructura organizativa de la LMF debería implementar algunas transformaciones radicales pero plausibles., Introduction: Children’s and youth football is a cultural and social phenomenon of huge importance in Argentina. Aim: This work aims at accounting for fundamental aspects of the organizational structure of children’s and youth football in the league of Mar del Plata, one of the most important cities in the country. Methods: The analysis of basic quantitative data enables us to account for the fundamental aspects of the organizational structure of children’s and youth football in the league of Mar del Plata. This elementary quantitative analysis is combined with an interpretation of reports based both on a discursive and an ethnographic analysis. Results & discussion: There is a severe process of exclusion and abandonment in the Mar del Plata football league (MFL). Not only it is the case that the number of female players is extremely low compared to the number of male players, but it is also the case that more than a thousand male players are abandoned year after year by the MFL. The complex process of abandonment and exclusion is also strongly connected with a series of diverse phenomena, like sexism, the absence of people with disabilities, the imitation of professional football practices, and the use of labels and stereotypes. Conclusions: Since it systematically promotes exclusion and abandonment, the MFL cannot fulfill an educational or inclusive function, and cannot even help to form high-level professional players. In its current state, the organizational structure of the MFL should apply some radical but plausible transformations., Resumo: Introdução: O futebol infantil e juvenil é um fenômeno cultural e social de grande importância na Argentina. Objetivo: Este trabalho visa dar conta dos aspectos fundamentais da estrutura organizacional do futebol infanto-juvenil da liga de Mar del Plata, uma das cidades mais importantes do país. Métodos: A análise de dados quantitativos básicos permite dar conta dos aspectos fundamentais da estrutura organizacional do futebol infanto-juvenil desta liga. Esta análise quantitativa elementar é combinada com uma interpretação de relatórios com base na análise discursiva e etnográfica. Resultados e discussão: Há um severo processo de exclusão e abandono no campeonato de futebol de Mar del Plata (LMF). Não só o número de jogadoras femininas é extremamente baixo em comparação com o número de jogadores masculinos, mas também acontece que mais de mil jogadores masculinos são abandonados ano após ano pela LMF. O complexo processo de abandono e exclusão também está fortemente ligado a uma série de fenômenos diversos, como o sexismo, a ausência de pessoas com deficiência, a imitação de práticas profissionais do futebol e o uso de rótulos e estereótipos. Conclusões: Por promover sistematicamente a exclusão e o abandono, o LMF não pode cumprir uma função educativa ou inclusiva e nem mesmo pode ajudar na formação de jogadores profissionais de alto nível. Em seu estado atual, a estrutura organizacional do MFL deve aplicar algumas transformações radicais, mas plausíveis.

The use of genomic and phenotypic data to scan for outliers is a mainstay for studies of hybridization and speciation. Geographic cline analysis of natural hybrid zones is widely used to identify putative signatures of selection by detecting deviations from baseline patterns of introgression. As with other outlier-based approaches, demographic histories can make neutral regions appear to be under selection and vice versa. In this study, we use a forward-time individual-based simulation approach to evaluate the robustness of geographic cline analysis under different evolutionary scenarios. We modelled multiple stepping-stone hybrid zones with distinct age, deme sizes, and migration rates, and evolving under different types of selection. We found that drift distorts cline shapes and increases false positive rates for signatures of selection. This effect increases with hybrid zone age, particularly if migration between demes is low. Drift can also distort the signature of deleterious effects of hybridization, with genetic incompatibilities and particularly underdominance prone to spurious typing as adaptive introgression. Our results suggest that geographic clines are most useful for outlier analysis in young hybrid zones with large populations of hybrid individuals. Current approaches may overestimate adaptive introgression and underestimate selection against maladaptive genotypes.

Background: Physical, chemical, bromatological and morphological characteristics are some of the quality aspects that regulate food products. The Andean blackberry is a very perishable fruit, due to its moisture, acidity, and water activity. Dehydration is one of the most common techniques in food preservation. Methods: In this research, blackberry concentrate, inulin and Lacticaseibacillus casei (L. casei), were co-encapsulated by spray drying using different conditions: maltodextrin (MD: 4-8%), inlet air temperature (IAT: 120-140 ºC), outlet air temperature (OAT: 70-80 ºC) and atomizing disc speed (ADS: 20000-24000 rpm). Results: The experimental optimization had a desirability function (71.9%), MD (5.0%), IAT (121.1 ºC), OAT (71.6 ºC) and ADS (24000 rpm). Physicochemical characteristics and antioxidant capacity were determined in spray dried product. The blackberry powder biofortified with L. casei and inulin (BPLI) achieved 7.12±0.69 Log CFU/g, total phenols (2912.9±96.4 mg eq-gallic acid/100g), anthocyanins (202.9±5.2 mg eq-cyd-3-glu/100g), and antioxidant capacity (ABTS: 2423.1±96.6 mg Trolox/100g and DPPH: 3302.6±76.4 mg Trolox/100g). Conclusions: The physicochemical properties of biofortified blackberry powder indicate that it is a product with appropriate solubility, humidity, and water activity for consumption and storage. BPLI has the potential to be a health food and could be positioned as a functional food due to its probiotic and prebiotic characteristics and the antioxidants that are beneficial to modern consumers; on the other hand, it represents a raw material with potential use in the food industry.

We report on national trends in STEM program quality using the Dimensions of Success (DoS), an empirical observation tool that provides a common definition of STEM program quality. We analyzed rati...

The authors have withdrawn their manuscript because they can no longer stand by the behavioral data set and corresponding manuscript as submitted to bioRxiv. A colleague downloaded the data and noted seven instances where pairs of values were duplicated across trials for different pairs of stimuli (data set with duplicated values highlighted is available as supplementary data). While omitting these values from the data set does not qualitatively change the conclusions presented, we have no means to validate the remaining data and therefore feel the best course of action is to treat the whole dataset as invalid. Consequently, analyses and conclusions based wholly or partly on behavioral data are invalid as well. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.

We revisit the merits of the retrospective pretest–posttest (RPP) design for repeated-measures research. The underutilized RPP method asks respondents to rate survey items twice during the same posttest measurement occasion from two specific frames of reference: “now” and “then.” Individuals first report their current attitudes or beliefs following a given intervention, and next they are prompted to think back to a specific time prior to the given intervention and rate the item again retrospectively. The design addresses many of the validity concerns that plague the traditional pretest–posttest design. Particularly when measuring noncognitive constructs, the RPP design allows participants to gauge the degree of change that they experience with greater awareness and precision than a traditional approach. We review the undesirable features of traditional designs and highlight the benefits of the retrospective approach. We offer examples from two recent, original studies and conclude with the recommendation that the RPP design be employed more broadly. We also conclude with a discussion of important directions for future examination of this design.

The objective of this study is to describe the characteristics of smokers admitted to different medical and surgical services in a university hospital and the perception of patients regarding the need for a specialized intervention. The sample comprises a total of 307 patients (mean age of 59.4 years), being 40% (n = 123) non-smokers, 42.7% (n = 131) ex-smokers, and 17.3% (n = 53) smokers. The average consumption of smokers was 22.2 cigarettes / day and the severity of nicotine dependence evaluated with the Fagerstrom test exceeded 5 points in more than half of the sample. On the other hand, 77.7% had made at least one previous attempt to quit tobacco use. Almost the entire sample (89.9 %) of smokers and ex-smokers considered it necessary to develop tobacco treatment programs during hospitalization. Finally, the importance of the hospital context is argued as an opportunity to address the cessation of smoking. The data obtained in this study will allow focusing more appropriately on the management of these patients and optimizing resources.

Darwin’s theory of sexual selection fundamentally changed how we think about sex and evolution. The struggle over mating and fertilization is a powerful driver of diversification within and among species. Contemporaries dismissed Darwin’s conjecture of a “taste for the beautiful” as favoring particular mates over others, but there is now overwhelming evidence for a primary role of both male and female mate choice in sexual selection. Darwin’s misogyny precluded much analysis of the “taste”; an increasing focus on mate choice mechanisms before, during, and after mating reveals that these often evolve in response to selection pressures that have little to do with sexual selection on chosen traits. Where traits and preferences do coevolve, they can do so whether fitness effects on choosers are positive, neutral, or negative. The spectrum of selection on traits and preferences, and how traits and preferences respond to social effects, determine how sexual selection and mate choice influence broader-scale processes like reproductive isolation and population responses to environmental change.

Understanding how organisms adapt to changing environments is a core focus of research in evolutionary biology. One common mechanism is adaptive introgression, which has received increasing attention as a potential route to rapid adaptation in populations struggling in the face of ecological change, particularly global climate change. However, hybridization can also result in deleterious genetic interactions that may limit the benefits of adaptive introgression. Here, we used a combination of genome-wide quantitative trait locus mapping and differential gene expression analyses between the swordtail fish species Xiphophorus malinche and X. birchmanni to study the consequences of hybridization on thermotolerance. While these two species are adapted to different thermal environments, we document a complicated architecture of thermotolerance in hybrids. We identify a region of the genome that contributes to reduced thermotolerance in individuals heterozygous for X. malinche and X. birchmanni ancestry, as well as widespread misexpression in hybrids of genes that respond to thermal stress in the parental species, particularly in the circadian clock pathway. We also show that a previously mapped hybrid incompatibility between X. malinche and X. birchmanni contributes to reduced thermotolerance in hybrids. Together, our results highlight the challenges of understanding the impact of hybridization on complex ecological traits and its potential impact on adaptive introgression.

BACKGROUND: Long-lasting insecticidal nets (LLINs) are currently the primary method of malaria control in sub-Saharan Africa and have contributed to a significant reduction in malaria burden over the past 15 years. However, this progress is threatened by the wide-scale selection of insecticide-resistant malaria vectors. It is, therefore, important to accelerate the generation of evidence for new classes of LLINs. METHODS: This protocol presents a three-arm superiority, single-blinded, cluster randomized controlled trial to evaluate the impact of 2 novel dual-active ingredient LLINs on epidemiological and entomological outcomes in Benin, a malaria-endemic area with highly pyrethroid-resistant vector populations. The study arms consist of (i) Royal Guard® LLIN, a net combining a pyrethroid (alpha-cypermethrin) plus an insect growth regulator (pyriproxyfen), which in the adult female is known to disrupt reproduction and egg fertility; (ii) Interceptor G2® LLIN, a net incorporating two adulticides (alpha-cypermethrin and chlorfenapyr) with different modes of action; and (iii) the control arm, Interceptor® LLIN, a pyrethroid (alpha-cypermethrin) only LLIN. In all arms, one net for every 2 people will be distributed to each household. Sixty clusters were identified and randomised 1:1:1 to each study arm. The primary outcome is malaria case incidence measured over 24 months through active case detection in a cohort of 25 children aged 6 months to 10 years, randomly selected from each cluster. Secondary outcomes include 1) malaria infection prevalence (all ages) and prevalence of moderate to severe anaemia in children under 5 years old, measured at 6 and 18 months post-intervention; 2) entomological indices measured every 3 months using human landing catches over 24 months. Insecticide resistance intensity will also be monitored over the study period. DISCUSSION: This study is the second cluster randomised controlled trial to evaluate the efficacy of these next-generation LLINs to control malaria transmitted by insecticide-resistant mosquitoes. The results of this study will form part of the WHO evidence-based review to support potential public health recommendations of these nets and shape malaria control strategies of sub-Saharan Africa for the next decade. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03931473 , registered on 30 April 2019.

Understanding how organisms adapt to changing environments is a core focus of research in evolutionary biology. One common mechanism is adaptive introgression, which has received increasing attention as a potential route to rapid adaptation in populations struggling in the face of ecological change, particularly global climate change. However, hybridization can also result in deleterious genetic interactions that may limit the benefits of adaptive introgression. Here, we used a combination of genome-wide quantitative trait locus mapping and differential gene expression analyses between the swordtail fish species Xiphophorus malinche and X. birchmanni to study the consequences of hybridization on thermotolerance. While these two species are adapted to different thermal environments, we document a complicated architecture of thermotolerance in hybrids. We identify a region of the genome that contributes to reduced thermotolerance in individuals heterozygous for X. malinche and X. birchmanni ancestry, as well as widespread misexpression in hybrids of genes that respond to thermal stress in the parental species, particularly in the circadian clock pathway. We also show that a previously mapped hybrid incompatibility between X. malinche and X. birchmanni contributes to reduced thermotolerance in hybrids. Together, our results highlight the challenges of understanding the impact of hybridization on complex ecological traits and its potential impact on adaptive introgression.

Pyrethroid resistance is widespread in sub-Saharan Africa. The objective of this study was to assess the insecticide resistance intensity in Anopheles gambiae s.l. (Diptera: Culicidae) in four districts of Benin in order to better understand how pyrethroid-only nets are likely to be effective. Thus, adult females of An. gambiae s.l., reared from field-collected larvae were used for assessing resistance intensity to permethrin and deltamethrin. They were tested at 1×, 5×, and 10× the diagnostic dose, using both WHO susceptibility tube testing and CDC bottle bioassays. Identification of molecular species, as well as of L1014F Kdr and Ace-1R mutations was performed using the PCR. The level of expression of biochemical enzymes was also evaluated. Overall, moderate to high resistance intensity to permethrin and deltamethrin was observed, irrespective of the testing method. While the L1014F Kdr frequency was high (>75%), Ace-1R was low (≤6%) in An. gambiaes.s. and Anopheles coluzzii, the two predominant species [52% (95% CI: 44.8–59.1) and 45% (95% CI: 38.0–52.2), respectively]. Anopheles arabiensis was found at very low frequency (3%, 95%CI: 1.1–6.4). For Biochemical analyses, α and β-esterases were over-expressed in all four districts, while mixed-function oxidases (MFOs) were over-expressed in only one. Overall, the two testing methods led to comparable conclusions, though there were a few inconsistencies between them. The moderate-high resistance intensity observed in the study area suggests that dual active-ingredient (AI) long-lasting insecticidal nets (LLINs) may provide better control of insecticide-resistant mosquitoes.

Bacteriophages are potent therapeutics against biohazardous bacteria, which rapidly develop multidrug resistance. However, routine administration of phage therapy is hampered by a lack of rapid production, safe bioengineering, and detailed characterization of phages. Thus, we demonstrate a comprehensive cell-free platform for personalized production, transient engineering, and proteomic characterization of a broad spectrum of phages. Using mass spectrometry, we validated hypothetical and non-structural proteins and could also monitor the protein expression during phage assembly. Notably, a few microliters of a one-pot reaction produced effective doses of phages against enteroaggregative Escherichia coli (EAEC), Yersinia pestis, and Klebsiella pneumoniae. By co-expressing suitable host factors, we could extend the range of cell-free production to phages targeting gram-positive bacteria. We further introduce a non-genomic phage engineering method, which adds functionalities for only one replication cycle. In summary, we expect this cell-free methodology to foster reverse and forward phage engineering and customized production of clinical-grade bacteriophages.

Photoacoustic (optoacoustic) imaging can extract molecular information with deeper tissue penetration than possible by fluorescence microscopy techniques. However, there is currently still a lack of robust genetically controlled contrast agents and molecular sensors that can dynamically detect biological analytes of interest with photoacoustics. In a biomimetic approach, we took inspiration from cuttlefish who can change their color by relocalizing pigment-filled organelles in so-called chromatophore cells under neurohumoral control. Analogously, we tested the use of melanophore cells from Xenopus laevis, containing compartments (melanosomes) filled with strongly absorbing melanin, as whole-cell sensors for optoacoustic imaging. Our results show that pigment relocalization in these cells, which is dependent on binding of a ligand of interest to a specific G protein-coupled receptor (GPCR), can be monitored in vitro and in vivo using photoacoustic mesoscopy. In addition to changes in the photoacoustic signal amplitudes, we could furthermore detect the melanosome aggregation process by a change in the frequency content of the photoacoustic signals. Using bioinspired engineering, we thus introduce a photoacoustic pigment relocalization sensor (PaPiReS) for molecular photoacoustic imaging of GPCR-mediated signaling molecules.

Apoptosis is a type of programmed cell death induced by a cascade of biochemical events, which leads to distinct morphological changes characterized by cell shrinkage, membrane blebbing, chromatin condensation, and DNA fragmentation. Apoptosis is executed by a class of cysteine proteases called caspases. Caspases are synthesized as inactive pro-caspases and activated by a series of cleavage reactions. Active caspases cleave cellular substrates and are thus the main effectors of the apoptotic cell death pathway. Detection of caspase cleavage by western blot analysis is a conventional method to demonstrate the induction of apoptosis. In the context of apoptosis, the proper analysis of western blot results depends on the understanding of the mechanisms and outcomes of caspase processing during the course of its activation. In this chapter, we describe the step-by-step methodology in the western blot analysis of caspase cleavage during apoptosis. We detail protocols for protein extraction, quantitation, casting, and running gel electrophoresis and western blot analysis of caspase -8 and caspase -9 activation. The described methods can be applied to any particular protein of interest.

Transcription factors orchestrate complex regulatory networks of gene expression. A better understanding of the common transcription factors, and their shared interactions, among a set of coregulated or differentially expressed genes can provide powerful insights into the key pathways governing such expression patterns. Critically, such information must also be considered in the context of the frequency in which a transcription factor is present in a properly selected background, and in the context of existing evidence of gene and transcription factor interaction. Given the vast amount of publicly available gene expression data that can be further scrutinized by the user-friendly analysis tools described here, many useful insights are assuredly to be revealed. The proceeding methods for application of the analysis tool CiiiDER for transcription factor-binding site identification, enrichment analysis, and coregulatory factor identification should be applicable to any dataset comparing differential gene expression in response to various stimuli and gene coexpression datasets. These methods should assist the researcher in identifying the most relevant regulators within a gene set, and refining the list of targets for future study to those which may share biologically important regulatory networks.

Within the cell, proteins are segregated into different organelles depending on their function and activation status. In response to stimulus, posttranslational modifications or loss of organelle membrane integrity lead to the movement of proteins from one compartment to another. This movement of proteins or protein translocation, exerts a significant effect on protein function. This is clearly demonstrated in the context of apoptosis wherein the cytoplasmic translocation of the mitochondrial resident protein, cytochrome C, initiates the activation of the intrinsic arm of the apoptotic pathway. Experimentally, protein translocation can be demonstrated by subcellular fractionation and subsequent western blot analysis of the isolated fractions. This chapter describes the step-by-step procedure in obtaining mitochondrial and cytoplasmic fractions from cell pellets and determining their purity and integrity.

The diversity of signalling traits within and across taxa is vast and striking, prompting us to consider how novelty evolves in the context of animal communication. Sexual selection contributes to diversification, and here we endeavour to understand the initial conditions that facilitate the maintenance or elimination of new sexual signals and receiver features. New sender and receiver variants can occur through mutation, plasticity, hybridization and cultural innovation, and the initial conditions of the sender, the receiver and the environment then dictate whether a novel cue becomes a signal. New features may arise in the sender, the receiver or both simultaneously. We contend that it may be easier than assumed to evolve new sexual signals because sexual signals may be arbitrary, sexual conflict is common and receivers are capable of perceiving much more of the world than just existing sexual signals. Additionally, changes in the signalling environment can approximate both signal and receiver changes through a change in transmission characteristics of a given environment or the use of new environments. The Anthropocene has led to wide-scale disruption of the environment and may thus generate opportunity to directly observe the evolution of new signals to address questions that are beyond the reach of phylogenetic approaches.

BACKGROUND: Nurse practitioners (NPs) play a critical role in the multidisciplinary management of patients with multiple sclerosis (MS). Neurologists´ behavioral characteristics have been associated with suboptimal clinical decisions. However, limited information is available on their impact among NPs involved in MS care. The aim of this study was to assess nurses´ therapeutic choices to understand behavioral factors influencing their decision making process. METHODS: A non-interventional, cross-sectional, web-based study was conducted. NPs actively involved in the care of patients with MS were invited to participate in the study by the Spanish Society of Neurology Nursing. Participants answered questions regarding their standard practice and therapeutic management of seven simulated relapsing-remitting MS (RRMS) case scenarios. A behavioral battery was used to measure participants´ life satisfaction, mood, positive social behaviors, feeling of helpfulness, attitudes toward adoption of evidence-based innovations, occupational burnout, and healthcare-related regret. The outcome of interest was therapeutic inertia (TI), defined as the lack of treatment escalation when there is clinical and radiological evidence of disease activity. A score to quantify TI was created based on the number of simulated scenarios where treatment intensification was warranted. RESULTS: Overall, 331 NPs were invited to participate, 130 initiated the study, and 96 (29%) completed the study. The mean age (SD) was 44.6 (9.8) years and 91.7% were female. Seventy-three participants (76.0%) felt their opinions had a significant influence on neurologists´ therapeutic decisions. Sixteen NPs (16.5%) showed severe emotional exhaustion related to work and 13 (13.5%) had depressive symptoms. The mean (SD) TI score was 0.97 (1.1). Fifty-six of NPs showed TI in at least one case scenario. Higher years of nursing experience (p = 0.014), feeling of helpfulness (p = 0.014), positive attitudes toward innovations (p = 0.046), and a higher intensity of care-related regret (p = 0.021) were associated with a lower risk of TI (adjusted R2 = 0.28). Burnout was associated with higher risk of TI (p = 0.001). CONCLUSIONS: Although NPs cannot prescribe MS treatments in Spain, their behavioral characteristics may influence the management of patients with RRMS. Continuing education and specific strategies for reducing occupational burnout may lead to better management skills and improve MS care.

The study of growth and possible metastasis in animal models of tumors would benefit from reliable cell labels for noninvasive whole-organism imaging techniques such as magnetic resonance imaging. Genetically encoded cell-tracking reporters have the advantage that they are contrast-selective for viable cells with intact protein expression machinery. Besides, these reporters do not suffer from dilution during cell division. Encapsulins, which are bacterial protein nanocompartments, can serve as genetically controlled labels for multimodal detection of cells. Such nanocompartments can host various guest molecules inside their lumen. These include, for example, fluorescent proteins or enzymes with ferroxidase activity leading to biomineralization of iron oxide inside the encapsulin nanoshell. The aim of this work was to implement heterologous expression of encapsulin systems from Quasibacillus thermotolerans using the fluorescent reporter protein mScarlet-I and ferroxidase IMEF in the human hepatocellular carcinoma cell line HepG2. The successful expression of self-assembled encapsulin nanocompartments with functional cargo proteins was confirmed by fluorescence microscopy and transmission electron microscopy. Also, coexpression of encapsulin nanoshells, ferroxidase cargo, and iron transporter led to an increase in T2-weighted contrast in magnetic resonance imaging of HepG2 cells. The results demonstrate that the encapsulin cargo system from Q. thermotolerans may be suitable for multimodal imaging of cancer cells and could contribute to further in vitro and in vivo studies.

Expression of exon-specific isoforms from alternatively spliced mRNA is a fundamental mechanism that substantially expands the proteome of a cell. However, conventional methods to assess alternative splicing are either consumptive and work-intensive or do not quantify isoform expression longitudinally at the protein level. Here, we therefore developed an exon-specific isoform expression reporter system (EXSISERS), which non-invasively reports the translation of exon-containing isoforms of endogenous genes by scarlessly excising reporter proteins from the nascent polypeptide chain through highly efficient, intein-mediated protein splicing. We applied EXSISERS to quantify the inclusion of the disease-associated exon 10 in microtubule-associated protein tau (MAPT) in patient-derived induced pluripotent stem cells and screened Cas13-based RNA-targeting effectors for isoform specificity. We also coupled cell survival to the inclusion of exon 18b of FOXP1, which is involved in maintaining pluripotency of embryonic stem cells, and confirmed that MBNL1 is a dominant factor for exon 18b exclusion. EXSISERS enables non-disruptive and multimodal monitoring of exon-specific isoform expression with high sensitivity and cellular resolution, and empowers high-throughput screening of exon-specific therapeutic interventions., Truong et al. developed a cell-based reporter system, EXSISERS, that enables non-invasive quantification of the protein expression levels of exon-specific isoforms via intein-mediated protein splicing.

Additional file 1. Information sheet and consent forms.

Background: Decisions based on erroneous assessments may result in unrealistic patient and family expectations, suboptimal advice, incorrect treatment, or costly medical errors. Regret is a common emotion in daily life that involves counterfactual thinking when considering alternative choices. Limited information is available on care-related regret affecting healthcare professionals managing patients with multiple sclerosis (MS). Methods: We reviewed identified gaps in the literature by searching for the combination of the following keywords in Pubmed: "regret and decision," "regret and physicians," and "regret and nurses." An expert panel of neurologists, a nurse, a psychiatrist, a pharmacist, and a psychometrics specialist participated in the study design. Care-related regret will be assessed by a behavioral battery including the standardized questionnaire Regret Intensity Scale (RIS-10) and 15 new specific items. Six items will evaluate regret in the most common social domains affecting individuals (financial, driving, sports-recreation, work, own health, and confidence in people). Another nine items will explore past and recent regret experiences in common situations experienced by healthcare professionals caring for patients with MS. We will also assess concomitant behavioral characteristics of healthcare professionals that could be associated with regret: coping strategies, life satisfaction, mood, positive social behaviors, occupational burnout, and tolerance to uncertainty. Planned Outcomes: This is the first comprehensive and standardized protocol to assess care-related regret and associated behavioral factors among healthcare professionals managing MS. These results will allow to understand and ameliorate regret in healthcare professionals. Spanish National Register (SL42129-20/598-E).

Bacteriophages are potent therapeutics against biohazardous bacteria, which rapidly develop multidrug resistance. However, routine administration of phage therapy is currently hampered by a lack of rapid phage production and efficient bioengineering and characterization capabilities. Thus, we have developed a comprehensive cell-free platform for time-efficient and personalized manufacturing, transient bioengineering, and proteomic characterization of a broad spectrum of phages. We validated 40 hypothetical proteins, including non-structural proteins, from T7 and CLB-P3 phages using high-resolution mass spectrometry and identified a new late-expressing gene cluster. Just a few microliters of the one-pot reaction produce effective doses of phages against enteroaggregative E. coli (EAEC) and potential biological warfare agents such as Yersinia pestis. By coexpressing suitable host factors, we could extend the range of cell-free phage production to include selective phages targeting gram-positive bacteria. We also introduce a transient, non-genomic phage engineering method, which adds functionalities for only one replication cycle. Finally, we showcase a pipeline for personalized phage therapy of a multidrug-resistant ESKAPE pathogen using a newly isolated and in-vitro produced phage against Klebsiella pneumoniae. We expect our cell-free methodology to enable accelerated reverse and forward phage engineering and the safe and customized production of clinical-grade therapeutic bacteriophages.

Supplementary Table 1: Select examples of novelty in mating communication as well as examples of mechanisms that may lead to novelty (noted as “NA (mechanism)” in the modality column). These examples and mechanisms are referenced in the manuscript.

Meters-long radio waves reveal a microscopic view of tissue ionic content., Despite its importance in regulating cellular or tissue function, electrical conductivity can only be visualized in tissue indirectly as voltage potentials using fluorescent techniques, or directly with radio waves. These either requires invasive procedures like genetic modification or suffers from limited resolution. Here, we introduce radio-frequency thermoacoustic mesoscopy (RThAM) for the noninvasive imaging of conductivity by exploiting the direct absorption of near-field ultrashort radio-frequency pulses to stimulate the emission of broadband ultrasound waves. Detection of ultrasound rather than radio waves enables micrometer-scale resolutions, over several millimeters of tissue depth. We confirm an imaging resolution of

The use of genomic and phenotypic data to scan for outliers is a mainstay for studies of hybridization and speciation. Geographic cline analysis of natural hybrid zones is widely used to identify putative signatures of selection by detecting deviations from baseline patterns of introgression. As with other outlier-based approaches, demographic histories can make neutral regions appear to be under selection and vice versa. In this study, we use a forward-time individual-based simulation approach to evaluate the robustness of geographic cline analysis under different evolutionary scenarios. We measured the effects of drift on genetic differentiation, and on false positive and false negatives detection using geographic clines. We modeled multiple stepping stone hybrid zones with distinct age, deme sizes, and migration rates, and evolving under different types of selection. We found that in young hybrid zones, drift increases overall genomic divergence, distorts cline shapes and increases both false positive and false negative rates. In old hybrid zones, genomic divergence and cline distortion are higher. Our results suggest that geographic clines are most useful for outlier analysis in young hybrid zones with large populations of hybrid individuals.

Loss mechanisms in fluid heating of cobalt ferrite (CFO) nanoparticles and CFO–Pd heterodimer colloidal suspensions are investigated as a function of particle size, fluid concentration and magnetic field amplitude. The specific absorption rate (SAR) is found to vary with increasing particle size due to a change in dominant heating mechanism from susceptibility to hysteresis and frictional loss. The maximum SAR is obtained for particle diameters of 11–15 nm as a result of synergistic contributions of susceptibility loss, including Néel and Brownian relaxation and especially hysteresis loss, thereby validating the applicability of linear response theory to superparamagnetic CFO nanoparticles. Our results show that the ferrofluid concentration and magnetic field amplitude alter interparticle interactions and associated heating efficiency. The SAR of the CFO nanoparticles could be maximized by adjusting the synthesis parameters. Despite the paramagnetic properties of individual palladium nanoparticles, CFO–Pd heterodimer suspensions were observed to have surprisingly improved magnetization as well as SAR values, when compared with CFO ferrofluids. This difference is attributed to interfacial interactions between the magnetic moments of paramagnetic Pd and superparamagnetic/ferrimagnetic CFO. SAR values measured from CFO–Pd heterodimer suspensions were found to be 47–52 W gFerrite−1, which is up to a factor of two higher than the SAR values of commercially available ferrofluids, demonstrating their potential as efficient heat mediators. Our results provide insight into the utilization of CFO–Pd heterodimer suspensions as potential nanoplatforms for diagnostic and therapeutic biomedical applications, e.g., in cancer hyperthermia, cryopreserved tissue warming, thermoablative therapy, drug delivery and bioimaging.

Biologists since Darwin have been fascinated by the evolution of sexually selected ornaments, particularly those that reduce viability. Uncovering the genetic architecture of these traits is key to understanding how they evolve and are maintained. Here, we investigate the genetic architecture of a sexually selected ornament, the “sword” fin extension that characterizes many species of swordtail fish (Xiphophorus). Using sworded and swordless sister species ofXiphophorus, we generated a mapping population and show that the sword ornament is polygenic – with ancestry across the genome explaining substantial variation in the trait. After accounting for the impacts of genome-wide ancestry, we identify one major effect QTL that explains ∼5% of the overall variation in the trait. Using a series of approaches, we narrow this large QTL interval to a handful of likely candidate genes, including the genesp8. Notably,sp8plays a regulatory role in fin regeneration and harbors several derived substitutions that are predicted to impact protein function in the species that has lost the sword ornament. Furthermore, we find evidence of selection on ancestry atsp8in four natural hybrid populations, consistent with selection against the sword in these populations.

Mapping vertebrate incompatibility alleles Deleterious gene interactions may underlie the observed hybrid incompatibilities. However, few genes underlying hybrid incompatibilities have been identified, and most of these involve species that do not hybridize in natural conditions. Powell et al. used genome sequencing to map genes likely responsible for incompatibilities that reduce fitness in naturally occurring hybrid swordtail fish. These gene combinations result in malignant melanoma, which is found in naturally hybridizing populations but is not present in the parental populations (see the Perspective by Dagilis and Matute). Using genome and population resequencing, the authors performed a genome-wide association study to identify potentially causative mutations. Using an admixture mapping approach that assessed introgression between multiple swordtail fish species, the authors suggest that lineages carry different genes that interact with the same candidate gene, resulting in the observed melanomas and providing insight into convergent hybrid incompatibles that arise between species. Science , this issue p. 731 ; see also p. 710

Background Many long noncoding RNAs (lncRNAs) have been implicated in general and cell type-specific molecular regulation. Here, we asked what underlies the fundamental basis for the seemingly random appearance of nuclear lncRNA condensates in cells, and we sought compounds that can promote the disintegration of lncRNA condensates in vivo. Results As a basis for comparing lncRNAs and cellular properties among different cell types, we screened lncRNAs in human pluripotent stem cells (hPSCs) that were differentiated to an atlas of cell lineages. We found that paraspeckles, which form by aggregation of the lncRNA NEAT1, are scaled by the size of the nucleus, and that small DNA-binding molecules promote the disintegration of paraspeckles and other lncRNA condensates. Furthermore, we found that paraspeckles regulate the differentiation of hPSCs. Conclusions Positive correlation between the size of the nucleus and the number of paraspeckles exist in numerous types of human cells. The tethering and structure of paraspeckles, as well as other lncRNAs, to the genome can be disrupted by small molecules that intercalate in DNA. The structure-function relationship of lncRNAs that regulates stem cell differentiation is likely to be determined by the dynamics of nucleus size and binding site accessibility. Graphical abstract

Behavior plays a fundamental role in shaping the origin and fate of species. Mating decisions can act to promote or restrict gene flow, as can personality traits that influence dispersal and niche use. Mate choice and personality are often both learned and therefore influenced by an individual's social environment throughout development. Likewise, the molecular pathways that shape these behaviors may also be co-expressed. In this study on swordtail fish (Xiphophorus birchmanni), we show that female mating preferences for species-typical pheromone cues are entirely dependent on social experience with adult males. Experience with adults also shapes development along the shy-bold personality axis, with shy behaviors arising from exposure to risk-averse heterospecifics as a potential stress-coping strategy. In maturing females, conspecific exposure results in a strong upregulation of olfaction and vision genes compared to heterospecific exposure, as well as immune response genes previously linked to anxiety, learning and memory. Conversely, heterospecific exposure involves an increased expression of genes important for neurogenesis, synaptic plasticity and social decision-making. We identify subsets of genes within the social decision-making network and with known stress-coping roles that may be directly coupled to the olfactory processes females rely on for social communication. Based on these results, we conclude that the social environment affects the neurogenomic trajectory through which socially-sensitive behaviors are learned, resulting in adult phenotypes adapted for specific social groupings.

Loss mechanisms in fluid heating of cobalt ferrite (CFO) nanoparticles and CFO-Pd heterodimer colloidal suspensions are investigated as a function of particle size, fluid concentration and magnetic field amplitude. The specific absorption rate (SAR) is found to vary with increasing particle size due to a change in dominant heating mechanism from susceptibility to hysteresis and frictional loss. The maximum SAR is obtained for particle diameters of 11-15 nm as a result of synergistic contributions of susceptibility loss, including Neel and Brownian relaxation and especially hysteresis loss, thereby validating the applicability of linear response theory to superparamagnetic CFO nanoparticles. Our results show that the ferrofluid concentration and magnetic field amplitude alter interparticle interactions and associated heating efficiency. The SAR of the CFO nanoparticles could be maximized by adjusting the synthesis parameters. Despite the paramagnetic properties of individual palladium nanoparticles, CFO-Pd heterodimer suspensions were observed to have surprisingly improved magnetization as well as SAR values, when compared with CFO ferrofluids. This difference is attributed to interfacial interactions between the magnetic moments of paramagnetic Pd and superparamagnetic/ferrimagnetic CFO. SAR values measured from CFO-Pd heterodimer suspensions were found to be 47-52 W gFerrite-1, which is up to a factor of two higher than the SAR values of commercially available ferrofluids, demonstrating their potential as efficient heat mediators. Our results provide insight into the utilization of CFO-Pd heterodimer suspensions as potential nanoplatforms for diagnostic and therapeutic biomedical applications, e.g., in cancer hyperthermia, cryopreserved tissue warming, thermoablative therapy, drug delivery and bioimaging.

An innovative system-building initiative known as the STEM Learning Ecosystems Community of Practice (SLECoP) is transforming U.S. STEM education through cross-sector partnerships between schools, afterschool and summer programs, libraries, museums, and businesses, among others. Although logic models exist to describe how SLEs can make positive contributions toward youth STEM learning in theory, it is unknown how individual SLEs are motivated or equipped to collect the evidence needed to demonstrate their value or abilities to solve the problems they were formed to address. The present study describes the results of a 34-item qualitative survey—completed by leaders of 37 SLEs from four U.S. regions—designed to understand where SLEs are in their evaluation planning, implementing, and capacity-building processes. We found that most SLEs were championed by the extended education sector, and all were highly motivated to conduct evaluation and assessment. Most communities reported a willingness to create a shared vision around data collection, which will help researchers and practitioners track, understand, and improve STEM quality and outcomes in and out of school.