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The politics of children's literature and the actors surrounding it have never been more visible than they are now, in the digital age. As one of the first children's series to gain widespread popularity concurrently with the spread of the internet, the Harry Potter septet arrived on the global stage at the perfect moment to develop an avid, connected fandom. But the fandom has laid bare the many conflicting ideologies of the fans themselves and of the actors surrounding the texts. This article examines the contentious issue of gender nonnormativity and its relation to the Harry Potter texts, the queer/trans reading practices and political resistance common to the fandom, and the ongoing disagreements over gender, made visible on social media, between Rowling and the fans of her series. The article discusses the Harry Potter novels' varied and conflicting ideologies; queer/trans readings of the Potter septet, including both invitations and resistances to queer/trans reading by Rowling herself; how gender is queered and queried in and through fan fiction; and finally, the recent hostilities between Rowling and her fans. It concludes by discussing the worsening relationship between Rowling and her fans and highlighting how fans are using their collective power to undermine Rowling's gender politics through fan fiction. By doing so, the article traces the complex politics of the reception of books for young people in the digital age, demonstrating that authors' powerful voices continue to shape readers' responses to texts long after their publication but showing, too, that readers often resist authors' attempts to influence not only their textual interpretations but their politics.

Abstract Peripheral inflammation, defined as inflammation that occurs outside the central nervous system, is an age-related phenomenon that has been identified as a risk factor for Alzheimer’s disease. While the role of chronic peripheral inflammation has been well characterized in the context of dementia and other age-related conditions, less is known about the neurologic contribution of acute inflammatory insults that take place outside the central nervous system. Herein, we define acute inflammatory insults as an immune challenge in the form of pathogen exposure (e.g., viral infection) or tissue damage (e.g., surgery) that causes a large, yet time-limited, inflammatory response. We provide an overview of the clinical and translational research that has examined the connection between acute inflammatory insults and Alzheimer’s disease, focusing on three categories of peripheral inflammatory insults that have received considerable attention in recent years: acute infection, critical illness, and surgery. Additionally, we review immune and neurobiological mechanisms which facilitate the neural response to acute inflammation and discuss the potential role of the blood–brain barrier and other components of the neuro-immune axis in Alzheimer’s disease. After highlighting the knowledge gaps in this area of research, we propose a roadmap to address methodological challenges, suboptimal study design, and paucity of transdisciplinary research efforts that have thus far limited our understanding of how pathogen- and damage-mediated inflammatory insults may contribute to Alzheimer’s disease. Finally, we discuss how therapeutic approaches designed to promote the resolution of inflammation may be used following acute inflammatory insults to preserve brain health and limit progression of neurodegenerative pathology.

Abstract Predicting COVID-19 severity is difficult, and the biological pathways involved are not fully understood. To approach this problem, we measured 4701 circulating human protein abundances in two independent cohorts totaling 986 individuals. We then trained prediction models including protein abundances and clinical risk factors to predict COVID-19 severity in 417 subjects and tested these models in a separate cohort of 569 individuals. For severe COVID-19, a baseline model including age and sex provided an area under the receiver operator curve (AUC) of 65% in the test cohort. Selecting 92 proteins from the 4701 unique protein abundances improved the AUC to 88% in the training cohort, which remained relatively stable in the testing cohort at 86%, suggesting good generalizability. Proteins selected from different COVID-19 severity were enriched for cytokine and cytokine receptors, but more than half of the enriched pathways were not immune-related. Taken together, these findings suggest that circulating proteins measured at early stages of disease progression are reasonably accurate predictors of COVID-19 severity. Further research is needed to understand how to incorporate protein measurement into clinical care.

Abstract Background Equine obesity is a significant health and welfare concern. The proportion of domestic horse populations that are overweight are as high as 45%. As the primary decision-makers for their horses’ care, owners are theoretically ideally placed to identify whether their horses are appropriately conditioned, however, research in other countries has shown that many owners are unable to accurately judge their horse’s body condition. In this study, through the comparison of body condition scoring (BCS) performed by an expert and the horse owners and interviews with owners, we aimed to identify the proportion of horses that were overweight or obese, to assess the accuracy of horse owners' BCS assessment both prior to and after receiving information and instructions on body condition scoring, and to identify common themes amongst owners’ views regarding BCS assessment and the Henneke BCS system. Results Forty-five percent of the horses in this study were overweight or obese. The agreement between the owners and an equine veterinarian regarding the horses’ BCS was fair to good both prior to (κ = .311, P

We investigate the binding of native β-cyclodextrin (β-CD) and eight novel β-CD derivatives with two different guest compounds, using isothermal calorimetry and 2D NOESY NMR. In all cases, the stoichiometry is 1:1 and binding is exothermic. Overall, modifications at the 3' position of β-CD, which is at the secondary face, weaken binding by several kJ/mol relative to native β-CD, while modifications at the 6' position (primary face) maintain or somewhat reduce the binding affinity. The variations in binding enthalpy are larger than the variations in binding free energy, so entropy-enthalpy compensation is observed. Characterization of the bound conformations with NOESY NMR shows that the polar groups of the guests may be situated at either face, depending on the host molecule, and, in some cases, both orientations are populated. The present results were used in the SAMPL7 blinded prediction challenge whose results are detailed in the same special issue of JCAMD.

Renewable fuels are needed to replace fossil fuels in the immediate future. Lignocellulosic bioenergy crops provide a renewable alternative that sequesters atmospheric carbon. To prevent displacement of food crops, it would be advantageous to grow biofuel crops on marginal lands. These lands will likely face more frequent and extreme drought conditions than conventional agricultural land, so it is crucial to see how proposed bioenergy crops fare under these conditions and how that may affect lignocellulosic biomass composition and saccharification properties. We found that while drought impacts the plant cell wall of Sorghum bicolor differently according to tissue and timing of drought induction, drought-induced cell wall compositional modifications are relatively minor and produce no negative effect on biomass conversion. This contrasts with the cell wall-related transcriptome, which had a varied range of highly variable genes (HVGs) within four cell wall-related GO categories, depending on the tissues surveyed and time of drought induction. Further, many HVGs had expression changes in which putative impacts were not seen in the physical cell wall or which were in opposition to their putative impacts. Interestingly, most pre-flowering drought-induced cell wall changes occurred in the leaf, with matrix and lignin compositional changes that did not persist after recovery from drought. Most measurable physical post-flowering cell wall changes occurred in the root, affecting mainly polysaccharide composition and cross-linking. This study couples transcriptomics to cell wall chemical analyses of a C4 grass experiencing progressive and differing drought stresses in the field. As such, we can analyze the cell wall-specific response to agriculturally relevant drought stresses on the transcriptomic level and see whether those changes translate to compositional or biomass conversion differences. Our results bolster the conclusion that drought stress does not substantially affect the cell wall composition of specific aerial and subterranean biomass nor impede enzymatic hydrolysis of leaf biomass, a positive result for biorefinery processes. Coupled with previously reported results on the root microbiome and rhizosphere and whole transcriptome analyses of this study, we can formulate and test hypotheses on individual gene candidates' function in mediating drought stress in the grass cell wall, as demonstrated in sorghum.

Advances in power electronics and their use in Miscellaneous Electric Loads (MELs) in buildings have resulted in increased interest in using low-voltage direct current (DC) power distribution as a replacement for the standard alternating current (AC) power distribution in buildings. Both systems require an endpoint converter to convert the distribution system voltage to the MELs voltage requirements. This study focused on the efficiency of these endpoint converters by testing pairs of AC/DC and DC/DC power converters powering the same load profile. In contrast to prior studies, which estimated losses based on data sheet efficiency and rated loads, in this study, we used part load data derived from real-world time-series load measurements of MELs and experimentally characterized efficiency curves for all converters. The measurements performed for this study showed no systematic efficiency advantage for commercially available DC/DC endpoint converters relative to comparable, commercially available AC/DC endpoint converters. For the eight appliances analyzed with the pair of converters tested, in 50%, the weighted energy efficiency of the DC/DC converter was higher, while, for the other 50%, the AC/DC converter was. Additionally, the measurements indicated that the common assumption of using either data sheet efficiency values or efficiency at full load may result in substantial mis-estimates of the system efficiency.

This mixed methods study, which is located in England investigates the capacity primary school teachers have to lay a foundation for pupils' engagement in active lifestyles through the delivery of quality Physical Education. The outlook is not positive as the study reveals low self-efficacy rates in the sample's ability (n181) to teach quality PE, a lack of consensus around what underpins positive outcomes in PE and limited prioritising of established pedagogical strategies deployed effectively in other curricular areas. Glimmers of hope and inspiration are drawn, however, from the best practice examples from 5 schools that provide evidence of a strong collective teacher efficacy towards PE. The study concludes PE is a force to mitigate health inequality with pragmatic suggestions offered to ensure primary teachers' sphere of influence is not wasted.

Our research community has limited understanding about the potential of video. This paper uses double stimulation and transformative agency to analyse two video modalities, extraction, and reflection; in a study about learning as it develops through family interactions. As researchers we were interested in children's motives. However, the children were young; therefore, posing a methodological problem. Drawing on video footage and 24 film elicitation interviews with children and fathers from 12 families in Norway, England, Hong Kong and Mumbai, India; we present case study examples using double stimulation to analyse video. We gathered between 5-10 hours of footage about father-child interactions from each family. To analyse video modalities through double stimulation, we identified the first stimulus, second stimulus, and manifestations of transformative agency. We found by combining double stimulation with video for the purposes of extraction, the non-visible becomes visible. The combination indicates opportunities for development that generate the kind of qualitative transformations, which imply child development can be seen in a new way. By combining double stimulation with video for reflection, researchers can trace outward affective and cognitive consequences of the 'mirror effect', back to children's conflict of motives, and how they decide to interpret what they see. Double stimulation therefore widens the potential of video. It shows video as an effective auxiliary stimulus to address the problem of researching young children's motives.

Purpose: The purpose of this paper is to explore the role of claims of cultural appropriation in negotiating who has the right to utilise specific racial, cultural or communities' ways of knowing in research co-production. Cultural appropriation is a claim made against those making illegitimate use of traditions, knowledge and practises that originate from specific racial and/or cultural group. Appropriation helps us interrogate the ways in which rewards and benefits in academia are distributed and shared, denied or hoarded, and by whom. Design/methodology/approach: The paper is a dialogue between two researchers in the in-betweens of research co-production, specifically in the negotiation of claiming the right to lead or engage in Afrofuturist work with communities. Findings: The claim of cultural appropriation is useful in naming, drawing boundaries and creating spaces for negotiation around access and ownership of academic work but must also develop as part of a broader transformative agenda towards racial equality in academia. Research limitations/implications: In addition to ethical considerations about power imbalances and extractive practises, research co-production should also be concerned with acknowledging and crediting knowledge production practises that originate from specific racial and/or cultural groups. Originality/value: This is the first paper to explore issues of cultural appropriation in research co-production, and co-production in relation to Afrofuturism. This extends ethical concerns on research co-production beyond academic power imbalances with, and extracting value from, communities to negotiating the relationships between academics and traditions, knowledge and practises developed by specific racial and/or cultural groups.

This paper presents an exploratory-interpretive study of two multilingual adults acquiring Norwegian through extensive reading. The study examined social and cognitive aspects of language acquisition, and individual factors, such as the language learning behaviors, experiences, attitudes, and beliefs of the participants. The data were collected using background self-reports, diaries in which the participants recorded their extensive reading and related language learning experiences, and semi-structured interviews. To represent adult language learning from a multilingual perspective, the data were analyzed qualitatively using a priori themes derived from the ecological model of multilinguality (Aronin, 2016; Aronin & Ó Laoire, 2004; Aronin & Singleton, 2012). The findings suggest that environment, previous education, reasons for learning a language, and previous knowledge of other languages affect how multilingual learners approach language learning and how they use a new language.

Background: There is limited literature evaluating patient satisfaction and outcomes after outpatient joint arthroplasty procedures in academic medical centers (AMCs). The aims of this study are to determine: (1) patients' desires to repeat their procedures and be discharged on the same day, (2) patient-reported outcome measures (PROMs), (3) time to discharge, (4) readmission rates, and (5) factors that predict PROMs in patients undergoing outpatient joint arthroplasty in AMCs. Methods: A prospective survey was completed by 66 total hip arthroplasty (THA), 35 total knee arthroplasty (TKA), and 180 unicondylar knee arthroplasty (UKA) outpatients who underwent surgery from May 2018 to December 2020 in 2 AMCs. The survey consisted of questions regarding hip or knee PROMs (Hip Disability and Osteoarthritis Outcome Score for Joint Replacement, and Knee Injury and Osteoarthritis Outcome Score for Joint Replacement), satisfaction with outpatient procedures and discharges, and reasons for readmissions. Linear regression analysis was conducted with statistical significance set at P < .05. Results: 100% of THA, 93.8% of TKA, and 93.0% of UKA outpatients stated that they would re-elect to undergo their respective procedure. Furthermore, 94% of THA, 81% of TKA, and 95% of UKA patients stated they would like same-day discharge again. THA, TKA, and UKA patients reported respective mean PROM scores of 94.7, 89.9, and 86.1. Readmission rates were 1.5%, 0.0%, and 0.5%, for THA, TKA, and UKA, respectively. Conclusions: Patients who underwent outpatient joint arthroplasty procedures at 2 AMCs experienced minimal readmissions and reported a high desire to repeat their outpatient procedures.

Objectives There is burgeoning evidence for the potential of nature-based interventions to improve wellbeing. However, the role of nature in enhancing maternal mental health, child development and early relationships remains relatively unexplored. This study aimed to develop a co-designed nature-based intervention to improve postnatal mothers’ and infants’ wellbeing.Design, setting and participants Person-based and co-design approaches informed the planning and design of the postnatal nature-based intervention. In stage 1, a multidisciplinary team was formed to agree research questions and appropriate methodology, and a scoping review was conducted. Six qualitative focus groups were then held with 30 mothers and 15 professional stakeholders. In stage 2, intervention guiding principles and a logic model were developed, and a stakeholder consensus meeting was convened to finalise the prototype intervention. The research was conducted in Bristol, UK, across voluntary, educational and community-based healthcare settings.Results Stakeholder consultation indicated significant enthusiasm for a postnatal nature-based intervention. A scoping review identified little existing research in this area. Focus group data are reported according to four broad themes: (1) perceived benefits of a group postnatal nature-based intervention, (2) potential drawbacks and barriers to access, (3) supporting attendance and implementation, and (4) ideas for intervention content. The developed intervention was tailored for mothers experiencing, or at risk of, postnatal mental health difficulties. It was identified that the intervention should facilitate engagement with the natural world through the senses, while taking into account a broad range of postnatal-specific practical and psychological needs.Conclusions To our knowledge, this is the first reported use of person-based and co-design approaches to develop a postnatal nature-based intervention. The resulting intervention was perceived by target users to address their needs and preferences. Further research is needed to determine the feasibility, clinical and cost-effectiveness of this approach.

Category: Midfoot/Forefoot; Other Introduction/Purpose: Minimally invasive surgery (MIS) is becoming increasingly popular among foot and ankle orthopaedic surgeons, as it has been shown to have limited complication rates and faster recovery times. Although there is robust data on MIS to treat hallux valgus, few studies have evaluated the role of MIS for 1st metatarsophalangeal (MTP) fusion to treat hallux rigidus - the most common arthritis of the foot. Hallux rigidus affects 2.5% of the U.S. population over age 50. Given this disease prevalence, it is important to characterize outcomes following MTP fusion procedures to optimize surgical approaches. In this retrospective cohort study, we aim to compare (1) patient-reported pain, (2) rate of successful fusion, and (3) rate of complications between groups who underwent open or MIS 1st MTP arthrodesis. Methods: This retrospective cohort study was conducted by evaluating clinical metrics, surgical outcomes, and radiographic data through chart review of 78 patients (25 male, 53 female), who underwent a 1st MTP fusion procedure at a large urban academic medical center from July 2015 through January 2023. 71 patients had a diagnosis of hallux rigidus, with other common diagnoses including hammertoe (24), hallux valgus (24), additional arthritides (11), and metatarsalgia (11). Two patients were excluded from our analysis due to lack of sufficient orthopaedic follow-up preventing evaluation of joint fusion. Pre- and post-operative pain scores were recorded on a 0-10 scale, though some notes documented subjective experience of pain (e.g., “minimal”). Complications were identified using post-operative clinic notes. The following radiographic data were included: hallux valgus angle, intermetatarsal angle, Coughlin classification, presence of arthritis, pre-operative sesamoid position, fusion construct materials, and success of fusion. Results: Of the 80 1st MTP fusion surgeries (two patients received both left and right foot operations), 51 were open and 29 were minimally invasive. There was no significant difference in the change in pre- to post-operative pain between the two groups (p=0.39). MIS patients had a higher successful fusion rate (100.0%) when compared to open surgery patients (94.1%), though this difference was insignificant (p=0.55). There was no significant difference in overall complication rates between MIS and open surgery groups (p=0.79). However, the MIS patients did not have any cases of deep infection or wound dehiscence and all (5/5) of their complications were related to hardware removal after fusion, whereas half of the complications (5/10) for the open surgery patients were related to infection and wounds. Conclusion: This retrospective analysis of 78 patients treated for hallux rigidus with 1st MTP fusion surgery showed no significant difference in rates of successful fusion and complications among patients who received an MIS procedure compared to an open surgery. There was no difference in patient-reported pain improvement between the two groups. Our main limitation is provider specialization, as only one surgeon in this study performed the MIS cases. Further studies should be done to obtain higher-level evidence to elucidate potential benefits of MIS over open surgery for 1st MTP arthrodesis.

Technology-enabled home-based cardiac rehabilitation (HBCR) is an emerging alternative to traditional center-based cardiac rehabilitation (CBCR), but little is known about outcomes in women. We analyzed 753 diverse and medically complex women who participated in HBCR and CBCR within an integrated health system and found both groups had similar clinical outcomes. Results suggest HBCR is a viable alternative to CBCR among women, including women with multiple comorbidities.

Background There are no validated clinical decision aids to identify neonates and young children at risk of hospital readmission or postdischarge mortality in sub-Saharan Africa, leaving the decision to discharge a child to a clinician’s impression. Our objective was to determine the precision of clinician impression to identify neonates and young children at risk for readmission and postdischarge mortality.Methods We conducted a survey study nested in a prospective observational cohort of neonates and children aged 1–59 months followed 60 days after hospital discharge from Muhimbili National Hospital in Dar es Salaam, Tanzania or John F. Kennedy Medical Center in Monrovia, Liberia. Clinicians who discharged each enrolled patient were surveyed to determine their perceived probability of the patient’s risk of 60-day hospital readmission or postdischarge mortality. We calculated the area under the precision-recall curve (AUPRC) to determine the precision of clinician impression for both outcomes.Results Of 4247 discharged patients, 3896 (91.7%) had available clinician surveys and 3847 (98.7%) had 60-day outcomes available: 187 (4.8%) were readmitted and 120 (3.1%) died within 60 days of hospital discharge. Clinician impression had poor precision in identifying neonates and young children at risk of hospital readmission (AUPRC: 0.06, 95% CI: 0.04 to 0.08) and postdischarge mortality (AUPRC: 0.05, 95% CI: 0.03 to 0.08). Patients for whom clinicians attributed inability to pay for future medical treatment as the reason for risk for unplanned hospital readmission had 4.76 times the odds hospital readmission (95% CI: 1.31 to 17.25, p=0.02).Conclusions Given the poor precision of clinician impression alone to identify neonates and young children at risk of hospital readmission and postdischarge mortality, validated clinical decision aids are needed to aid in the identification of young children at risk for these outcomes.

ABSTRACTAs humanity pushes deeper into the Anthropocene, Social-Ecological Systems (SESs) across the world are facing mounting pressures. Managing, protecting and understanding these systems require research into their complex and interlinked nature. One area that has been met with increased research in recent times is Sense of Place (SoP), broadly defined as the emotional bond that people have with a ‘place’. There has been substantial growth in the number of studies seeking to understand and quantify SoP across different contexts to help integrate it into decision-making processes. This targeted scoping review aims to explore the environmental literature for examples of applied studies that measure SoP in social-ecological contexts with a focus on how SoP has been quantified and measured. Our results show a growing body of literature as well as a number of clear knowledge gaps. The majority of studies to date have focussed on the Global North (accounting for 79% of the published papers), particularly the USA and Australia, with the distribution of authors closely mirroring this same pattern. Marine and freshwater ecosystems were understudied in comparison to terrestrial ecosystems. Mixed methods were most commonly used to collect data, with interviews and surveys being the most common instruments. Further research into why SoP is being measured is required, but preliminary investigations indicate there is a perceived applicability to policy and planning. We explore these findings and identify research directions.

ABSTRACTBackground Adults with chronic pain have a lower quality of life (QOL) compared to the general population. Chronic pain requires specialized treatment to address the multitude of factors that contribute to an individual’s pain experience, and effectively managing pain requires a biopsychosocial approach to improve patients’ QOL.Aim This study examined adults with chronic pain after a year of specialized treatment to determine the role of cognitive markers (i.e., pain catastrophizing, depression, pain self-efficacy) in predicting changes in QOL.Methods Patients in an interdisciplinary chronic pain clinic (N = 197) completed measures of pain catastrophizing, depression, pain self-efficacy, and QOL at baseline and 1 year later. Correlations and a moderated mediation were completed to understand the relationships between the variables.Results Higher baseline pain catastrophizing was significantly associated with increased mental QOL (b = 0.39, 95% confidence interval [CI] 0.141; 0.648) and decreased depression (b = −0.18, 95% CI −0.306; −0.052) over a year. Furthermore, the relationship between baseline pain catastrophizing and the change in depression was moderated by the change in pain self-efficacy (b = −0.10, 95% CI −0.145; −0.043) over a year. Patients with high baseline pain catastrophizing reported decreased depression after a year of treatment, which was associated with greater QOL improvements but only in patients with unchanged or improved pain self-efficacy.Conclusions Our findings highlight the roles of cognitive and affective factors and their impact on QOL in adults with chronic pain. Understanding the psychological factors that predict increased mental QOL is clinically useful, because medical teams can optimize these positive changes in QOL through psychosocial interventions aimed at improving patients’ pain self-efficacy.

Environmental scientists are acutely aware of the increasing dangers posed by the climate crisis, and this professional awareness is linked to raised levels of climate anxiety. In this paper we explore the use of group therapy as a tool to create a safe space for researchers to share their feelings on climate change. We examine the transcripts of a 2-day group therapy session provided to seven environmental scientists based in the United States by a professional therapist. We analyse more than 12 hours of anonymised audio transcripts to identify patterns, observations and shared experiences. Our results suggest that group therapy may provide positive and cathartic experiences for environmental scientists through sharing emotions and experiences with peers, both about the challenges of their professional lives and difficulties in processing feelings about their work subjects and the climate crisis. Further, results indicate that participants benefited from sharing strategies for coping with the emotional toll of the climate crisis.

The Covid-19 pandemic had wide-ranging effects on how people lived, worked and learned. Consequently, electricity use was altered from pre-pandemic patterns. The deviation from expected electricity usage patterns in commercial properties due to the Covid-19 pandemic was analyzed in a medium-sized American city. The focus on a single community (1) allows usage to be linked specifically to the timing of public health and executive orders; and (2) provides a fine-grained, detailed understanding of usage in different property classifications (e.g. restaurants, hotels, schools, outpatient medical facilities, offices, and religious organizations). Electricity consumption data from 2019, adjusted for average daily temperature, were used to calculate expected use in 2020. Electricity usage was found to be lower than expected for most commercial property classes, but the timing and magnitude of these effects varied. For example, within the hospitality industry, hotels evidenced a larger and more sustained decrease in usage (–17%) as compared with restaurants (–11%). In addition, usage patterns for outpatient medical facilities can be linked to specific executive orders. Together, a heterogenous rate of electricity use is found to vary according to specific types of commercial properties. Practice relevance In much of the research regarding electricity usage, utility customers are placed into the broad categories of residential, commercial and industrial. These groups are viewed as homogeneous. This study examined different property classes within the commercial group during the Covid-19 pandemic. Although electricity usage was lower than expected for most commercial property classes, the timing and magnitude of these effects varied. For example, within the hospitality industry, hotels evidenced a larger and more sustained decrease in usage than did restaurants. In addition, usage patterns for outpatient medical facilities were linked to specific executive orders. Together, these data sources allowed for a detailed examination of Covid-19’s effect on electricity use for specific types of commercial properties and it revealed that their response to the pandemic varied widely. Thus, it is an oversimplification to view commercial properties as a single, homogeneous group.

Summary: Extracellular vesicles (EVs) can mediate intercellular communication, including signaling between the placenta and maternal tissues. Human placental explant culture is a versatile in vitro model system to investigate placental function. We performed systematic studies in different tissue culture media types and oxygen tensions to identify a defined serum-free culture condition that supports high trophoblast viability and metabolism, as well as the release of similar populations of EVs, compared to traditional undefined conditions that contain media additives potentially contaminated with exogenous EVs. We also determined the time frame in which trophoblast viability and functionality remain optimal. Multiplex vesicle flow cytometry with classical EV and placenta-specific markers revealed three separate populations of explant-derived EVs: small CD63+ EVs; large PLAP+ EVs; and CD63-/PLAP- EVs. These culture and analytical approaches will enable in vitro modeling of short-term effects of environmental perturbations associated with pregnancy complications on placental function and EV release.

Adenosine deaminase (ADA) is a human mononuclear Zn2+ metalloenzyme that converts adenosine to inosine. ADA is a validated drug target for cancer, but there has been little recent work on the development of new therapeutics against this enzyme. The lack of new advancements can be partially attributed to an absence of suitable assays for high-throughput screening (HTS) against ADA. To facilitate more rapid drug discovery efforts for this target, an in vitro assay was developed that utilizes the enzymatic conversion of a visibly emitting adenosine analogue to the corresponding fluorescent inosine analogue by ADA, which can be monitored via fluorescence intensity changes. Utilizing this assay, a library of ∼350 small molecules containing metal-binding pharmacophores (MBPs) was screened in an HTS format to identify new inhibitor scaffolds against ADA. This approach yielded a new metal-binding scaffold with a Ki value of 26±1 μM.

Next-generation sequencing studies of saliva and dental plaque from subjects in both healthy and diseased states have identified bacteria belonging to the Rothia genus as ubiquitous members of the oral microbiota. To gain a deeper understanding of molecular mechanisms underlying the chemical ecology of this unexplored group, we applied a genome mining approach that targets functionally important biosynthetic gene clusters (BGCs). All 45 genomes that were mined, representing Rothia mucilaginosa, Rothia dentocariosa, and Rothia aeria, harbored a catechol-siderophore-like BGC. To explore siderophore production further, we grew the previously characterized R. mucilaginosa ATCC 25296 in liquid cultures, amended with glycerol, which led to the identification of the archetype siderophore enterobactin by using tandem liquid chromatography-mass spectrometry (LC-MS/MS), high-performance liquid chromatography (HPLC), and nuclear magnetic resonance (NMR) spectroscopy. Normally attributed to pathogenic gut bacteria, R. mucilaginosa is the first commensal oral bacterium found to produce enterobactin. Cocultivation studies including R. mucilaginosa or purified enterobactin revealed that enterobactin reduced growth of certain strains of cariogenic Streptococcus mutans and pathogenic strains of Staphylococcus aureus Commensal oral bacteria were either unaffected, reduced in growth, or induced to grow adjacent to enterobactin-producing R. mucilaginosa or the pure compound. Taken together with Rothia's known capacity to ferment a variety of carbohydrates and amino acids, our findings of enterobactin production add an additional level of explanation to R. mucilaginosa's prevalence in the oral cavity. Enterobactin is the strongest Fe(III) binding siderophore known, and its role in oral health requires further investigation.IMPORTANCE The communication language of the human oral microbiota is vastly underexplored. However, a few studies have shown that specialized small molecules encoded by BGCs have critical roles such as in colonization resistance against pathogens and quorum sensing. Here, by using a genome mining approach in combination with compound screening of growth cultures, we identified that the commensal oral community member R. mucilaginosa harbors a catecholate-siderophore BGC, which is responsible for the biosynthesis of enterobactin. The iron-scavenging role of enterobactin is known to have positive effects on the host's iron pool and negative effects on host immune function; however, its role in oral health remains unexplored. R. mucilaginosa was previously identified as an abundant community member in cystic fibrosis, where bacterial iron cycling plays a major role in virulence development. With respect to iron's broad biological importance, iron-chelating enterobactin may explain R. mucilaginosa's colonization success in both health and disease.

Marine cyanobacteria (blue-green algae) have been shown to possess an enormous capacity to produce structurally diverse natural products that exhibit a broad spectrum of potent biological activities, including cytotoxic, antifungal, antiparasitic, antiviral, and antibacterial activities. Using mass-spectrometry-guided fractionation together with molecular networking, cyanobacterial field collections from American Samoa and Palmyra Atoll yielded three new cyclic peptides, tutuilamides A-C. Their structures were established by spectroscopic techniques including 1D and 2D NMR, HR-MS, and chemical derivatization. Structure elucidation was facilitated by employing advanced NMR techniques including nonuniform sampling in combination with the 1,1-ADEQUATE experiment. These cyclic peptides are characterized by the presence of several unusual residues including 3-amino-6-hydroxy-2-piperidone and 2-amino-2-butenoic acid, together with a novel vinyl chloride-containing residue. Tutuilamides A-C show potent elastase inhibitory activity together with moderate potency in H-460 lung cancer cell cytotoxicity assays. The binding mode to elastase was analyzed by X-ray crystallography revealing a reversible binding mode similar to the natural product lyngbyastatin 7. The presence of an additional hydrogen bond with the amino acid backbone of the flexible side chain of tutuilamide A, compared to lyngbyastatin 7, facilitates its stabilization in the elastase binding pocket and possibly explains its enhanced inhibitory potency.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

This report describes the first application of the novel NMR-based machine learning tool "Small Molecule Accurate Recognition Technology" (SMART 2.0) for mixture analysis and subsequent accelerated discovery and characterization of new natural products. The concept was applied to the extract of a filamentous marine cyanobacterium known to be a prolific producer of cytotoxic natural products. This environmental Symploca extract was roughly fractionated, and then prioritized and guided by cancer cell cytotoxicity, NMR-based SMART 2.0, and MS2-based molecular networking. This led to the isolation and rapid identification of a new chimeric swinholide-like macrolide, symplocolide A, as well as the annotation of swinholide A, samholides A-I, and several new derivatives. The planar structure of symplocolide A was confirmed to be a structural hybrid between swinholide A and luminaolide B by 1D/2D NMR and LC-MS2 analysis. A second example applies SMART 2.0 to the characterization of structurally novel cyclic peptides, and compares this approach to the recently appearing "atomic sort" method. This study exemplifies the revolutionary potential of combined traditional and deep learning-assisted analytical approaches to overcome longstanding challenges in natural products drug discovery.

With the potential development of new disease-modifying Alzheimer's disease (AD) therapies, simple, widely available screening tests are needed to identify which individuals, who are experiencing symptoms of cognitive or behavioral decline, should be further evaluated for initiation of treatment. A blood-based test for AD would be a less invasive and less expensive screening tool than the currently approved cerebrospinal fluid or amyloid β positron emission tomography (PET) diagnostic tests. We examined whether plasma tau phosphorylated at residue 181 (pTau181) could differentiate between clinically diagnosed or autopsy-confirmed AD and frontotemporal lobar degeneration. Plasma pTau181 concentrations were increased by 3.5-fold in AD compared to controls and differentiated AD from both clinically diagnosed (receiver operating characteristic area under the curve of 0.894) and autopsy-confirmed frontotemporal lobar degeneration (area under the curve of 0.878). Plasma pTau181 identified individuals who were amyloid β-PET-positive regardless of clinical diagnosis and correlated with cortical tau protein deposition measured by 18F-flortaucipir PET. Plasma pTau181 may be useful to screen for tau pathology associated with AD.

Abstract: The discovery of biologically active small molecules requires sifting through large amounts of data to identify unique or unusual arrangements of atoms. Here, we develop, test and evaluate an atom‐based sort to identify novel features of secondary metabolites and demonstrate its use to evaluate novelty in marine microbial and sponge extracts. This study outlines an important ongoing advance towards the translation of autonomous systems to identify, and ultimately elucidate, atomic novelty within a complex mixture of small molecules.