Estimating biotic integrity to capture existence value of freshwater ecosystems.

The US Environmental Protection Agency (EPA) uses a water quality index (WQI) to estimate benefits of proposed Clean Water Act regulations. The WQI is relevant to human use value, such as recreation, but may not fully capture aspects of nonuse value, such as existence value. Here, we identify an index of biological integrity to supplement the WQI in a forthcoming national stated preference survey that seeks to capture existence value of streams and lakes more accurately within the conterminous United States (CONUS). We used literature and focus group research to evaluate aquatic indices regularly reported by the EPA's National Aquatic Resource Surveys. We chose an index that quantifies loss in biodiversity as the observed-to-expected (O/E) ratio of taxonomic composition because focus group participants easily understood its meaning and the environmental changes that would result in incremental improvements. However, available datasets of this index do not provide the spatial coverage to account for how conditions near survey respondents affect their willingness to pay for its improvement. Therefore, we modeled and interpolated the values of this index from sampled sites to 1.1 million stream segments and 297,071 lakes across the CONUS to provide the required coverage. The models explained 13 to 36% of the variation in O/E scores and demonstrate how modeling can provide data at the required density for benefits estimation. We close by discussing future work to improve performance of the models and to link biological condition with water quality and habitat models that will allow us to forecast changes resulting from regulatory options.

Is a randomised controlled trial of take home naloxone distributed in emergency settings likely to be feasible and acceptable? Findings from a UK qualitative study exploring perspectives of people who use opioids and emergency services staff.

OBJECTIVE: Distribution of take-home naloxone (THN) by emergency services may increase access to THN and reduce deaths and morbidity from opioid overdose. As part of a feasibility study for a randomised controlled trial (RCT) of distribution of THN kits and education within ambulance services and Emergency Departments (EDs), we used qualitative methods to explore key stakeholders' perceptions of feasibility and acceptability of delivering the trial. METHODS: We undertook semi-structured interviews and focus groups with 26 people who use opioids and with 20 paramedics and ED staff from two intervention sites between 2019 and 2021. Interviews and focus groups were recorded, transcribed verbatim and analysed using Framework Analysis. RESULTS: People using opioids reported high awareness of overdose management, including personal experience of THN use. Staff perceived emergency service provision of THN as a low-cost, low-risk intervention with potential to reduce mortality, morbidity and health service use. Staff understood the trial aims and considered it compatible with their work. All participants supported widening access to THN but reported limited trial recruitment opportunities partly due to difficulties in consenting patients during overdose. Procedural problems, restrictive recruitment protocols, limited staff buy-in and patients already owning THN limited trial recruitment. Determining trial effectiveness was challenging due to high levels of alternative community provision of THN. CONCLUSIONS: Distribution of THN in emergency settings was considered feasible and acceptable for stakeholders but an RCT to establish the effectiveness of THN delivery is unlikely to generate further useful evidence due to difficulties in recruiting patients and assessing benefits.

Intuitive cooperators: Time pressure increases children's cooperative decisions in a modified public goods game.

There is mounting empirical evidence to suggest that adults are intuitively cooperative. When presented with a cooperative dilemma between self-maximizing and benefitting the common good, decisions made quickly are more likely to be cooperative, whereas slow decisions tend to favor self-interest. To investigate the ontogenetic origins of intuitive cooperation, we examined the development of intuitive cooperation in middle childhood. We presented 150 children (7-12 years of age) with an online child-friendly public goods game where participants had a choice between giving two resources to themselves or four to their group. Participants were assigned to one of three decision time conditions; speeded, neutral, or delayed. We found that when decisions were speeded, children were more likely to cooperate compared to when decisions were unconstrained or delayed. Furthermore, children's intuitive choices only favored cooperation if they believed their peers were also cooperative. This pattern of findings held across the age range included in this study. Our findings suggest that in middle and late childhood, children are intuitively cooperative when making decisions to benefit the common good. HIGHLIGHTS: Time pressure increases children's cooperation in a public goods game, compared to when decisions are delayed or unconstrained. Between 7 and 12 years of age children engage in costly cooperation most of the time regardless of decision time. When children believe others are generally cooperative, their intuition is to cooperate. From middle to late childhood, intuitive decisions favor costly cooperation towards the common good.

Two- Versus 8-Zone Lung Ultrasound in Heart Failure: Analysis of a Large Data Set Using a Deep Learning Algorithm.

OBJECTIVE: Scanning protocols for lung ultrasound often include 8 or more lung zones, which may limit real-world clinical use. We sought to compare a 2-zone, anterior-superior thoracic ultrasound protocol for B-line artifact detection with an 8-zone approach in patients with known or suspected heart failure using a deep learning (DL) algorithm. METHODS: Adult patients with suspected heart failure and B-lines on initial lung ultrasound were enrolled in a prospective observational study. Subjects received daily ultrasounds with a hand-held ultrasound system using an 8-zone protocol (right and left anterior/lateral and superior/inferior). A previously published deep learning algorithm that rates severity of B-lines on a 0-4 scale was adapted for use on hand-held ultrasound full video loops. Average severities for 8 and 2 zones were calculated utilizing DL ratings. Bland-Altman plot analyses were used to assess agreement and identify bias between 2- and 8-zone scores for both primary (all patients, 5728 videos, 205 subjects) and subgroup (confirmed diagnosis of heart failure or pulmonary edema, 4464 videos, 147 subjects) analyses. RESULTS: Bland-Altman plot analyses revealed excellent agreement for both primary and subgroup analyses. The absolute difference on the 4-point scale between 8- and 2-zone average scores was not significant for the primary dataset (0.03; 95% CI -0.01 to 0.07) or the subgroup (0.01; 95% CI -0.04 to 0.06). CONCLUSION: Utilization of a 2-zone, anterior-superior thoracic ultrasound protocol provided similar severity information to an 8-zone approach for a dataset of subjects with known or suspected heart failure.

Predator Versus Pathogen: How Does Predatory Bdellovibrio bacteriovorus Interface with the Challenges of Killing Gram-Negative Pathogens in a Host Setting?

Bdellovibrio bacteriovorus is a small deltaproteobacterial predator that has evolved to invade, reseal, kill, and digest other gram-negative bacteria in soils and water environments. It has a broad host range and kills many antibiotic-resistant, clinical pathogens in vitro, a potentially useful capability if it could be translated to a clinical setting. We review relevant mechanisms of B. bacteriovorus predation and the physiological properties that would influence its survival in a mammalian host. Bacterial pathogens increasingly display conventional antibiotic resistance by expressing and varying surface and soluble biomolecules. Predators coevolved alongside prey bacteria and so encode diverse predatory enzymes that are hard for pathogens to resist by simple mutation. Predators do not replicate outside pathogens and thus express few transport proteins and thus few surface epitopes for host immune recognition. We explain these features, relating them to the potential of predatory bacteria as cellular medicines.

Invasion of the body snatchers: the role of parasite introduction in host distribution and response to salinity in invaded estuaries.

In dynamic systems, organisms are faced with variable selective forces that may impose trade-offs. In estuaries, salinity is a strong driver of organismal diversity, while parasites shape species distributions and demography. We tested for trade-offs between low-salinity stress and parasitism in an invasive castrating parasite and its mud crab host along salinity gradients of two North Carolina rivers. We performed field surveys every six to eight weeks over 3 years to determine factors influencing parasite prevalence, host abundance, and associated taxa diversity. We also looked for signatures of low-salinity stress in the host by examining its response (time-to-right and gene expression) to salinity. We found salinity and temperature significantly affected parasite prevalence, with low-salinity sites (less than 10 practical salinity units (PSU)) lacking infection, and populations in moderate salinities at warmer temperatures reaching prevalence as high as 60%. Host abundance was negatively associated with parasite prevalence. Host gene expression was plastic to acclimation salinity, but several osmoregulatory and immune-related genes demonstrated source-dependent salinity response. We identified a genetic marker that was strongly associated with salinity against a backdrop of no neutral genetic structure, suggesting possible selection on standing variation. Our study illuminates how selective trade-offs in naturally dynamic systems may shape host evolutionary ecology.

The mitochondrial proteins NLRX1 and TUFM form a complex that regulates type I interferon and autophagy.

The mitochondrial protein MAVS (also known as IPS-1, VISA, and CARDIF) interacts with RIG-I-like receptors (RLRs) to induce type I interferon (IFN-I). NLRX1 is a mitochondrial nucleotide-binding, leucine-rich repeats (NLR)-containing protein that attenuates MAVS-RLR signaling. Using Nlrx1(-/-) cells, we confirmed that NLRX1 attenuated IFN-I production, but additionally promoted autophagy during viral infection. This dual function of NLRX1 paralleled the previously described functions of the autophagy-related proteins Atg5-Atg12, but NLRX1 did not associate with Atg5-Atg12. High-throughput quantitative mass spectrometry and endogenous protein-protein interaction revealed an NLRX1-interacting partner, mitochondrial Tu translation elongation factor (TUFM). TUFM interacted with Atg5-Atg12 and Atg16L1 and has similar functions as NLRX1 by inhibiting RLR-induced IFN-I but promoting autophagy. In the absence of NLRX1, increased IFN-I and decreased autophagy provide an advantage for host defense against vesicular stomatitis virus. This study establishes a link between an NLR protein and the viral-induced autophagic machinery via an intermediary partner, TUFM.

NLRX1 protein attenuates inflammatory responses to infection by interfering with the RIG-I-MAVS and TRAF6-NF-κB signaling pathways.

The nucleotide-binding domain and leucine-rich-repeat-containing (NLR) proteins regulate innate immunity. Although the positive regulatory impact of NLRs is clear, their inhibitory roles are not well defined. We showed that Nlrx1(-/-) mice exhibited increased expression of antiviral signaling molecules IFN-ß, STAT2, OAS1, and IL-6 after influenza virus infection. Consistent with increased inflammation, Nlrx1(-/-) mice exhibited marked morbidity and histopathology. Infection of these mice with an influenza strain that carries a mutated NS-1 protein, which normally prevents IFN induction by interaction with RNA and the intracellular RNA sensor RIG-I, further exacerbated IL-6 and type I IFN signaling. NLRX1 also weakened cytokine responses to the 2009 H1N1 pandemic influenza virus in human cells. Mechanistically, Nlrx1 deletion led to constitutive interaction of MAVS and RIG-I. Additionally, an inhibitory function is identified for NLRX1 during LPS activation of macrophages where the MAVS-RIG-I pathway was not involved. NLRX1 interacts with TRAF6 and inhibits NF-κB activation. Thus, NLRX1 functions as a checkpoint of overzealous inflammation.

Guiding Cardiopulmonary Resuscitation with Focused Echocardiography: A Report of Five Cases.

Background: Focused transthoracic echocardiography has been used to determine etiologies of cardiac arrest and evaluate utility of continuing resuscitation after cardiac arrest. Few guidelines exist advising ultrasound timing within the advanced cardiac life support algorithm. Natural timing of echocardiography occurs during the pulse check, when views are unencumbered by stabilization equipment or vigorous movements. However, recent studies suggest that ultrasound performance during pulse checks prolongs the pause duration of cardiopulmonary resuscitation. Transesophageal echocardiography studies have demonstrated benefits in this regard, but there have been no transthoracic echocardiography studies assessing the physical performance of compressions during cardiopulmonary resuscitation. Objective: The purpose of this study was to describe cases where echocardiography performed at the beginning of the cardiac arrest algorithm offers actionable information to cardiopulmonary resuscitation itself without delaying provision of compressions. Conclusion: Providers using focused echocardiography to evaluate cardiac arrest patients should consider initiating scans at the start of compressions to identify the optimal location for compression delivery and to detect inadequate compressions. Subsequent visualization of full left ventricular compression may be seen after a location change, and combined with end tidal carbon dioxide values, gives indication for improved forward circulatory flow. Although it is not possible in all patients, doing so hastens provision of quality compressions that affect hemodynamic parameters without causing prolongations to the pulse check pause. Further research is needed to determine patient outcomes from both out-of-hospital and in-hospital cardiac arrest when cardiopulmonary resuscitation is visually guided by focused echocardiography.

Valuing Hemlock Woolly Adelgid Control in Public Forests: Scope Effects with Attribute Non-Attendance.

Sensitivity to the scope of public good provision is an important indication of validity for the contingent valuation method. An online survey was administered to an opt-in non-probability sample panel to estimate the willingness-to-pay to protect hemlock trees from a destructive invasive species on federal land in North Carolina. We collected survey responses from 907 North Carolina residents. We find evidence that attribute non-attendance (ANA) is a factor when testing for sensitivity to scope. When estimating the model with stated ANA, the ecologically and socially important scope coefficients become positive and statistically significant with economically significant marginal willingness-to-pay estimates.

An Integrated Assessment Model for Valuing Water Quality Changes in the U.S.

The US Environmental Protection Agency (EPA) often requires expertise from environmental assessors, hydrologists, economists, and others to analyze the benefits of regional and national policy decisions related to changes in water quality. This led EPA to develop two models to form an Integrated Assessment Model (IAM): HAWQS is a web-based water quantity and quality modeling systems and BenSPLASH is a modeling platform for quantifying the economic benefits of changes in water quality. This paper discusses the development of the component models and applies HAWQS and BenSPLASH to a case study in the Republican River Basin.

Integrating perspectives: How the development of second-personal competence lays the foundation for a second-personal morality.

The integration of first-, second-, and third-personal information within joint intentional collaboration provides the foundation for broad-based second-personal morality. We offer two additions to this framework: a description of the developmental process through which second-personal competence emerges from early triadic interactions, and empirical evidence that collaboration with a concrete goal may provide an essential focal point for this integrative process.

The NLRP3 inflammasome mediates in vivo innate immunity to influenza A virus through recognition of viral RNA.

The nucleotide-binding domain and leucine-rich-repeat-containing (NLR) family of pattern-recognition molecules mediate host immunity to various pathogenic stimuli. However, in vivo evidence for the involvement of NLR proteins in viral sensing has not been widely investigated and remains controversial. As a test of the physiologic role of the NLR molecule NLRP3 during RNA viral infection, we explored the in vivo role of NLRP3 inflammasome components during influenza virus infection. Mice lacking Nlrp3, Pycard, or caspase-1, but not Nlrc4, exhibited dramatically increased mortality and a reduced immune response after exposure to the influenza virus. Utilizing analogs of dsRNA (poly(I:C)) and ssRNA (ssRNA40), we demonstrated that an NLRP3-mediated response could be activated by RNA species. Mechanistically, NLRP3 inflammasome activation by the influenza virus was dependent on lysosomal maturation and reactive oxygen species (ROS). Inhibition of ROS induction eliminated IL-1beta production in animals during influenza infection. Together, these data place the NLRP3 inflammasome as an essential component in host defense against influenza infection through the sensing of viral RNA.

A gene trap transposon eliminates haematopoietic expression of zebrafish Gfi1aa, but does not interfere with haematopoiesis.

A transposon-mediated gene trap screen identified the zebrafish line qmc551 that expresses a GFP reporter in primitive erythrocytes and also in haemogenic endothelial cells, which give rise to haematopoietic stem and progenitor cells (HSPCs) that seed sites of larval and adult haematopoiesis. The transposon that mediates this GFP expression is located in intron 1 of the gfi1aa gene, one of three zebrafish paralogs that encode transcriptional repressors homologous to mammalian Gfi1 and Gfi1b proteins. In qmc551 transgenics, GFP expression is under the control of the endogenous gfi1aa promoter, recapitulates early gfi1aa expression and allows live observation of gfi1aa promoter activity. While the transposon integration interferes with the expression of gfi1aa mRNA in haematopoietic cells, homozygous qmc551 fish are viable and fertile, and display normal primitive and definitive haematopoiesis. Retained expression of Gfi1b in primitive erythrocytes and up-regulation of Gfi1ab at the onset of definitive haematopoiesis in homozygous qmc551 carriers, are sufficient to allow normal haematopoiesis. This finding contradicts previously published morpholino data that suggested an essential role for zebrafish Gfi1aa in primitive erythropoiesis.

Regulation of mitochondrial antiviral signaling pathways.

Mitochondrial antiviral immunity involves the detection of viral RNA by intracellular pattern-recognition receptors (PRRs) belonging to the RIG-I-like helicase family. The convergence of these and other signaling molecules to the outer mitochondrial membrane results in the rapid induction of antiviral cytokines including type-1 interferon. Here, we discuss recent studies describing new molecules implicated in the regulation of this antiviral response.

Preschoolers' generosity increases with understanding of the affective benefits of sharing.

Research has demonstrated that sharing with others is rewarding, suggesting a proximal mechanism of humans' extraordinary tendency to engage in prosocial behavior. The current study explored the cognitive basis of the relation between generosity and happiness early in ontogeny. We demonstrate that preschool children understand the relation between generosity and happiness. Moreover, our results show that children's emotion ratings are predictive for their subsequent sharing behavior. This finding provides evidence for the theoretical claim that prosocial behavior may be related to the anticipation of positive feelings, and that this mechanism may explain early instances of generosity in preschool children.

Drosophila MUS312 and the vertebrate ortholog BTBD12 interact with DNA structure-specific endonucleases in DNA repair and recombination.

DNA recombination and repair pathways require structure-specific endonucleases to process DNA structures that include forks, flaps, and Holliday junctions. Previously, we determined that the Drosophila MEI-9-ERCC1 endonuclease interacts with the MUS312 protein to produce meiotic crossovers, and that MUS312 has a MEI-9-independent role in interstrand crosslink (ICL) repair. The importance of MUS312 to pathways crucial for maintaining genomic stability in Drosophila prompted us to search for orthologs in other organisms. Based on sequence, expression pattern, conserved protein-protein interactions, and ICL repair function, we determined that the mammalian ortholog of MUS312 is BTBD12. Orthology between these proteins and S. cerevisiae Slx4 helped identify a conserved interaction with a second structure-specific endonuclease, SLX1. Genetic and biochemical evidence described here and in related papers suggest that MUS312 and BTBD12 direct Holliday junction resolution by at least two distinct endonucleases in different recombination and repair contexts.

Affiliation affects generosity in young children: The roles of minimal group membership and shared interests.

Young children's willingness to share with others is selective, and is affected by their level of affiliation with the recipients of their generosity. We explored affiliation's impact on sharing behavior with two experiments comparing the effects of two distinct affiliative cues-minimal group membership and shared interests. Children (4- to 6-year-olds) completed a resource allocation task, making forced-choice decisions as to how to distribute stickers between themselves and others. In Experiment 1, the sharing partners were minimal in- and out-group members; in Experiment 2, they differed in their opinion of the participants' interests. Both experiments' manipulations affected feelings of affiliation, as indicated by children's stated friendship preferences and perceptions of similarity. More notably, both minimal group membership and interests affected sharing behavior. Children made fewer generous allocations toward out-group members than toward in-group members. Similarly, children made fewer generous allocations when recipients disliked their interests than when recipients shared those interests or when their opinions were unknown. Across experiments, the recipient manipulations' effects on generosity were similar in their pattern and magnitude despite fundamental differences between the two affiliative cues. These findings highlight the broad impact of affiliation on young children's sharing behavior.

Toddlers Involve Their Caregiver to Help Another Person in Need.

Three experiments with 18- to 35-month-old children (n = 169) studied toddler-caregiver interactions when being confronted with another person in need. In particular, we explored whether toddlers would request their caregiver to help a needy other when they are not able to help themselves. Children observed another person who needed help to accomplish a task, but were either not able to provide help as the object was out of reach (Experiment 1) or because an obstacle prevented children from interacting with the other person (Experiments 2, 2b). The experiments revealed the same developmental trend with 2.5-year-olds selectively involving their caregiver to help the needy other. The results are interpreted in terms of toddlers' motivation to see others helped and with respect to their developing ability to actively involve others to regulate their emotions.