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Abstract The Gulf and Atlantic Coastal Plains of the southern United States are characterized by a wide continental shelf that was subaerially exposed for ca. 80,000 years during glacial-interval marine regressions and transgressions. Given their present submergence, little is known about the vegetation dynamics, particularly at annual time scales, of these formerly terrestrial sites due to erosional processes associated with marine transgressions. Here, we present an annually resolved and well-replicated 489-year tree-ring chronology from macrobotanical specimens—anatomically identified as Taxodium distichum (L.) Rich.—collected in situ from a recently exposed submerged forest in 18 m water depth in the northern Gulf of Mexico. This chronology not only reveals historical vegetation dynamics at annual resolutions during Marine Isotope Stages 3–5a, but it also captures a catastrophic mortality event likely connected to intense storm activity, perhaps driven by freshwater fluxes from Heinrich events. Our findings are supported by coupled climate model simulations from the last glaciation, providing new insights into the environmental history of the southeastern US coastal regions.

Abstract The distribution and abundance of ectothermic mosquitoes are strongly affected by temperature, but mechanisms remain unexplored. We describe the effect of temperature on the transcriptome of Anopheles stephensi, an invasive vector of human malaria. Adult females were maintained across a range of mean temperatures (20 °C, 24 °C and 28 °C), with daily fluctuations of +5 °C and −4 °C at each mean temperature. Transcriptomes were described up to 19 days post-blood meal. Of the >3100 differentially expressed genes, we observed shared temporal expression profiles across all temperatures, suggesting their indispensability to mosquito life history. Tolerance to 20 and 28 ( + 5°C/−4°C) was associated with larger and more diverse transcriptomes compared to 24 ( + 5 °C/−4 °C). Finally, we identified two distinct trends in gene expression in response to blood meal ingestion, oxidative stress, and reproduction. Our work has implications for mosquitoes’ response to thermal variation, mosquito immune-physiology, mosquito-malaria interactions and the development of vector control tools.

Abstract Background Rotational vertebral artery (VA) syndrome represents a rare mechanical vasculopathy that can lead to vertebrobasilar insufficiency and ischemic stroke. Objective We describe a 10‐year‐old boy with a history of chronic morning emesis who presented with acute onset dizziness, gait instability, and vomiting and was diagnosed with acute ischemic posterior circulation stroke. Interpretation Contrast‐enhanced magnetic resonance angiography (MRA) of the head and neck with dynamic positioning demonstrated loss of flow‐related enhancement with head tilted to the left, conferring a diagnosis of rotational VA syndrome. He started aspirin monotherapy and subsequently underwent C1 laminectomy, with both radiographic and clinical improvement on follow‐up. Conclusion Dynamic contrasted‐enhanced MRA imaging can serve as a noninvasive alternative to digital subtraction angiography in the diagnosis of rotational VA syndrome and should be considered in suspected cases of pediatric rotational arteriopathy.

IntroductionChildren and adolescents with attention-deficit/hyperactivity disorder (ADHD) are more likely to report a lifetime history of concussion compared to those without ADHD. Prior studies have predominantly relied on surveys of youth who self-reported their ADHD status (as opposed to parent report), examined samples with relatively narrow age ranges, and focused on youth athletes. It is unclear if ADHD severity or ADHD medication influences the association between ADHD and greater lifetime history of concussion. We investigated the association between parent-reported ADHD, ADHD severity, and medication status with parent-reported lifetime concussion history in children and adolescents ranging in age from 5 to 17, sampled from the United States general population.MethodsParent reported health data from 33,914 children and adolescents were selected from the 2021 National Survey of Children’s Health. Lifetime history of concussion, current ADHD diagnosed by a healthcare provider, and, for those with current ADHD, ADHD severity (mild or moderate/severe), and current ADHD medication status (yes/no) were reported by parents. A Mantel–Haenszel analysis was used to assess the associations between ADHD and lifetime concussion history across five age groups (ages 5–7, 8–10, 11–13, 14–15, and 16–17). Additional Mantel–Haenszel analyses were used to assess the associations between lifetime history of concussion and ADHD severity as well as medication status across five age groups.ResultsOverall, 6.2% of youth had a parent-reported history of concussion and 12.3% had parent-reported current ADHD. Twice as many youth with ADHD (10.6%) had a parent-reported history of concussion compared to youth without ADHD (5.6%). There were no differences in lifetime concussion history for those with mild (10.9%) versus moderate/severe ADHD (10.3%). Similarly, among those with ADHD, there were no differences in lifetime concussion history for those taking medication (9.9%) versus those not taking medication (11.6%).DiscussionConsistent with previous literature, youth with ADHD had greater lifetime concussion history than youth without ADHD (approximately twice the prevalence in those with ADHD). Contrary to our hypotheses among youth with ADHD, parent-reported ADHD severity and current medication status were not associated with lifetime concussion history.

ABSTRACT The widespread prevalence of Asaia in mosquitoes makes it a potential candidate for paratrangenic control in Anopheles. To better understand whether this bacterium could be used for malaria control, we quantified Asaia in An. gambiae s.l populations in malaria endemic regions examining co-infection with Plasmodium falciparum. Adult Anopheles mosquitoes were collected across two different eco-geographical localities in Cameroon, during both the dry and wet seasons. DNA was extracted from whole individual mosquitoes, and real time-qPCR amplification of the 16S ribosomal RNA was used to quantify Asaia in both An. gambiae and An. coluzzii samples. We also detected and quantified P. falciparum infection in the same mosquitoes. The density of Asaia was successfully quantified in a total of 864 field mosquitoes, comprising of 439 An. gambiae from Bankeng and 424 An. coluzii collected from Gounougou. Interestingly, a higher prevalence of Asaia in An. gambiae (88.3%) compared to An. coluzzii (80.9%) was observed. Moreover, the density of Asaia in both species was significantly affected by seasonal changes in the two localities. Furthermore, a significant difference between the infection densities of Asaia and the Plasmodium infection status in the two species was recorded. However, no correlation was observed between the number of Asaia and P. falciparum infections. This study provides evidence that naturally occurring Asaia infection is not correlated to P. falciparum development within An. gambiae and An. coluzzii. Nevertheless, further studies incorporating experimental infections are required to better investigate the correlation between Anopheles mosquitoes, Asaia, and Plasmodium.IMPORTANCEThe symbiont Asaia has emerged as a promising candidate for paratransgenic control of malaria, but further analysis of its biology and genetics across Africa is necessary. In this study, we investigated and quantified the influence of Asaia in naturally infected An. gambiae s.l. populations with the malaria parasite Plasmodium falciparum. Genomic DNA was extracted from whole individual mosquitoes collected from two localities, and Asaia was quantified using real-time qPCR by amplification of the 16S ribosomal RNA gene. We also detected and quantified Plasmodium falciparum infection in the same mosquitoes and established the correlation between Asaia and Plasmodium coinfection. This study provides evidence that naturally occurring Asaia infection is not correlated with P. falciparum development within An. gambiae and An. coluzzii mosquitoes.

Background: Rugby league is a popular collision sport among Australian adolescent and young adult men. Concussion is one of the more common injuries in rugby league. Few studies have examined concussion in youth rugby league. To examine medically diagnosed concussions from a single season within two elite-level pathway rugby league competitions by evaluating game play risk factors and conducting a video review of potential concussion signs. Methods: All players involved in the Queensland Rugby League's (QRL) under 18 years and under 20 years age group competitions during the 2019 season were included in this study. Data included all head injury assessments (HIAs) identified in real-time through the QRL injury surveillance system for these two QRL age group competitions. The purpose of this study was to (i) report the rates of HIAs and medically diagnosed concussions; (ii) examine video signs of potential concussion; (iii) review game play risk factors related to HIAs and concussions; and (iv) determine the number of days until a concussed player returned to match play and the number of subsequent games missed by concussed players. Results: There were 86 HIAs and 30 medically diagnosed concussions from the two competitions. The concussion incidence was 2.93 per 1000 player match hours in the under 18-year age group and 5.75 per 1000 player match hours in the under 20-year age group. Slow to stand was the most commonly observed video sign (78.6%; 22/28 concussions). Most concussed players (91%, 21/23) missed at least one subsequent game (M ​= ​1.4, SD ​= ​1.7, range ​= ​0–7 games), with the average days to return-to-play being 15.7 (SD ​= ​7.0, range ​= ​7–41 days). Conclusions: In elite-level pathway rugby league, the incidences of HIAs and medically diagnosed concussions were higher in the under 20 age group than the under 18 age group. Both age groups had lower incidences of HIAs and concussions than professional adult rugby league players. Return-to-play following concussion was similar across the two age groups and differed considerably compared to the elite level, with a longer time before return to play for the younger elite level development pathway players.

Mosquitoes transmit medically important human pathogens, including viruses like dengue virus and parasites such as Plasmodium spp., the causative agent of malaria. Mosquito microbiomes are critically important for the ability of mosquitoes to transmit disease-causing agents. However, while large collections of bacterial isolates and genomic data exist for vertebrate microbiomes, the vast majority of work in mosquitoes to date is based on 16S rRNA gene amplicon data that provides limited taxonomic resolution and no functional information. To address this gap and facilitate future studies using experimental microbiome manipulations, we generated a bacterial Mosquito-Associated Isolate Collection (MosAIC) consisting of 392 bacterial isolates with extensive metadata and high-quality draft genome assemblies that are publicly available, both isolates and sequence data, for use by the scientific community. MosAIC encompasses 142 species spanning 29 bacterial families, with members of the Enterobacteriaceae comprising 40% of the collection. Phylogenomic analysis of 3 genera, Enterobacter, Serratia, and Elizabethkingia, reveal lineages of mosquito-associated bacteria isolated from different mosquito species in multiple laboratories. Investigation into species' pangenomes further reveals clusters of genes specific to these lineages, which are of interest for future work to test for functions connected to mosquito host association. Altogether, we describe the generation of a physical collection of mosquito-associated bacterial isolates, their genomic data, and analyses of selected groups in context of genome data from closely related isolates, providing a unique, highly valuable resource for research on bacterial colonisation and adaptation within mosquito hosts. Future efforts will expand the collection to include broader geographic and host species representation, especially from individuals collected from field populations, as well as other mosquito-associated microbes, including fungi, archaea, and protozoa.

IntroductionAttention-deficit/hyperactivity disorder (ADHD) is a neurodevelopmental disorder characterized by attention problems, excessive activity, and impulsivity - occurring in approximately 13% of children 12–17 years of age, and as many as 16% of older adolescents 18–22 years of age, with a greater prevalence in boys than girls. ADHD frequently co-occurs with specific learning disorder (LD), a neurodevelopmental disorder characterized by difficulties learning and using academic skills, such as reading, writing, and mathematics. Taken together, ADHD and/or LD are common among high school students and college students and can influence performance on computerized tests used in concussion management. However, normative data for widely used computer-based measures used in concussion management do not include reference samples with ADHD and/or LD. Previous research has documented the expected frequency of obtaining low scores on computer-based neuropsychological test measures among healthy, uninjured individuals, but few studies have examined the frequency of obtaining low scores in athletes with neurodevelopmental diagnoses, such as ADHD and/or LD. This study examined the frequency of low scores (i.e., multivariate base rates) on the Immediate Post-Concussion Assessment and Cognitive Test (ImPACT) for youth and young adults with self-reported ADHD and/or LD.MethodsThe sample included 174,878 adolescent and young adult student-athletes who completed pre-season baseline neurocognitive assessments, who were assigned to the following independent groups: ADHD only (n = 45,215), LD only (n = 54,223), ADHD and LD (n = 45,737), No ADHD or LD (n = 47,684; “control athletes”).ResultsBase rates of low scores were stratified by high school (e.g., 13–18) and collegiate age (e.g., 19–22) and sex. It was common for student athletes (all ages) with LD alone (i.e., 30–37%), or with both ADHD and LD (i.e., 24–31%), to obtain at least two low ImPACT composite scores, but not three low composite scores. However, it was relatively uncommon for control athletes (those without ADHD or LD) (i.e., 12–14%) or older athletes (ages 19–22) with ADHD (i.e., 14–15%) to obtain two (or more) low scores.DiscussionHaving information relating to the base rates of low scores among uninjured athletes enhances the interpretation of ImPACT results among the broader population of student-athletes with and without neurodevelopmental disorders.

Concussions in contact sports are challenging for athletes, health professionals and sporting bodies to prevent, detect and manage. Design of interventions for primary prevention, early recognition of concussion and continuing to improve postconcussion management are essential for protecting athletes and promoting brain health. Over the last decade, there have been advancements in video technology for analysing head impact events and improvements in the clinical management of concussions. This study protocol describes how researchers, clinicians and staff from the Australasian National Rugby League (NRL) have brought these advancements together and developed a database of videos with head impact events and clinical outcomes. The intended outputs from this work will enhance the understanding of head impact events in NRL, from biomechanical and gameplay factors to concussion and return to play outcomes. Publishing this protocol increases the transparency of this large-scale effort to better identify head impacts and their relationship to concussions and player movement behaviour to contextualise these variables to generate new knowledge and support the reproducibility of these emerging findings. Between 2017 and 2023, over 5250 head contact cases were recorded in the database, from which >1700 head injury assessments were performed, and >600 concussions were diagnosed. Future studies using these data are planned to inform both primary and secondary injury prevention initiatives, such as risk analysis and prediction of game scenarios that result in concussion, as well as investigation of features and factors that help to inform the duration of recovery and return to play.

There are no validated diagnostic criteria for traumatic encephalopathy syndrome (TES). During the early and middle 20th century, TES was described as a clinical condition that was experienced by some high-exposure boxers?and it was believed to reflect chronic traumatic brain injury. Consensus criteria for the diagnosis of TES were published in 2021. We applied the consensus criteria for TES retrospectively to cases of chronic brain damage in boxers described in articles published in the 20th century that were obtained from narrative and systematic reviews. The sample included 157 boxers identified in 21 articles published between 1929 and 1999. Two authors reviewed each case description and coded the criteria for TES. For the core clinical features, cognitive impairment was noted in 63.1%, and in 28.7% of cases the person's cognitive functioning appeared to be broadly normal. Neurobehavioral dysregulation was present in 25.5%. One third (34.4%) were identified as progressive, 30.6% were not progressive, and the course could not be clearly determined in 35.0%. In total, 29.9% met the TES consensus criteria, 28.0% did not, and 42.0% had insufficient information to make a diagnostic determination. TES, in the 20th century, was described as a neurological condition, not a psychiatric disorder?and this supports the decision of the 2021 consensus group to remove primary and secondary psychiatric diagnoses from being a core diagnostic feature. Future research is needed to determine whether, or the extent to which, cognitive impairment or neurobehavioral dysregulation described as characterizing TES are associated with chronic traumatic encephalopathy neuropathological change.

Abstract Background The rugby league tackle has been identified as the game event with the greatest propensity for a clinically diagnosed concussion. This study aims to replicate the work conducted in professional rugby league and rugby union by examining Head Injury Assessment (HIA) events to determine the associated tackle characteristics that increase concussion risk in sub-elite rugby league players. This comparison between competition levels is important due to the less developed physiological and tackle proficiency characteristics of sub-elite rugby league players and the fewer resources available for an on-field diagnosis, compared to the elite level of the sport. Results Tackles resulting in Head Injury Assessments (HIAs, n = 131) and 2,088 tackles that did not result in a head injury were identified and coded from one season of the 2019 Queensland Cup. The body position of both ball carrier and tackler, tackle height, and body contact areas were evaluated. The propensity for tacklers to undergo a head injury assessment was 1.49 HIAs per 1,000 tackles, equating to a 2.5-fold higher risk than that of the ball carrier (0.59 HIAs per 1,000 tackles). The risk for an HIA was 2.75-fold greater when the tackler was upright (2.89 HIAs per 1,000 tackles) compared to a bent-at-the-waist tackler (1.05 HIAs per 1,000 tackles). The greatest risk for the tackler and ball carrier sustaining an HIA occurred when the tackle height was high, with head-to-head contact having the greatest propensity for an HIA (44.37 HIAs per 1,000 tackles). HIA risk was also greater for both players when the ball carrier did not employ an evasion strategy (3.73 HIAs per 1,000 tackles). Conclusions The study replicates results from research in elite rugby league and rugby union. A combination of higher head contact/proximity and upright body position significantly increase an HIA risk. Tackler head position and ball carrier evasion behaviours also affect risk, suggesting that injury prevention strategies designed to reduce tackle height and improve tackle technique by focusing on head position, body position, and in a novel finding, ball carrier evasion, may reduce head injury risk in sub-elite rugby league players.

The brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.), is an important vector for Rickettsia rickettsii, causative agent of Rocky Mountain spotted fever. Current public health prevention and control efforts to protect people involve preventing tick infestations on domestic animals and in and around houses. Primary prevention tools rely on acaricides, often synthetic pyrethroids (SPs); resistance to this chemical class is widespread in ticks and other arthropods. Rhipicephalus sanguineus s.l. is a complex that likely contains multiple unique species and although the distribution of this complex is global, there are differences in morphology, ecology, and perhaps vector competence among these major lineages. Two major lineages within Rh. sanguineus s.l., commonly referred to as temperate and tropical, have been documented from multiple locations in North America, but are thought to occupy different ecological niches. To evaluate potential acaricide resistance and better define the distributions of the tropical and temperate lineages throughout the US and in northern Mexico, we employed a highly multiplexed amplicon sequencing approach to characterize sequence diversity at: 1) three loci within the voltage-gated sodium channel (VGSC) gene, which contains numerous genetic mutations associated with resistance to SPs; 2) a region of the gamma-aminobutyric acid-gated chloride channel gene (GABA-Cl) containing several mutations associated with dieldrin/fipronil resistance in other species; and 3) three mitochondrial genes (COI, 12S, and 16S). We utilized a geographically diverse set of Rh sanguineus s.l. collected from domestic pets in the US in 2013 and a smaller set of ticks collected from canines in Baja California, Mexico in 2021. We determined that a single nucleotide polymorphism (T2134C) in domain III segment 6 of the VGSC, which has previously been associated with SP resistance in Rh. sanguineus s.l., was widespread and abundant in tropical lineage ticks (>50 %) but absent from the temperate lineage, suggesting that resistance to SPs may be common in the tropical lineage. We found evidence of multiple copies of GABA-Cl in ticks from both lineages, with some copies containing mutations associated with fipronil resistance in other species, but the effects of these patterns on fipronil resistance in Rh. sanguineus s.l. are currently unknown. The tropical lineage was abundant and geographically widespread, accounting for 79 % of analyzed ticks and present at 13/14 collection sites. The temperate and tropical lineages co-occurred in four US states, and as far north as New York. None of the ticks we examined were positive for Rickettsia rickettsii or Rickettsia massiliae.

Abstract Over recent decades, the southeastern United States (Southeast) has become increasingly well represented by the terrestrial climate proxy record. However, while the paleo proxy records capture the region's hydroclimatic history over the last several centuries, the understanding of near surface air temperature variability is confined to the comparatively shorter observational period (1895‐present). Here, we detail the application of blue intensity (BI) methods on a network of tree‐ring collections and examine their utility for producing robust paleotemperature estimates. Results indicate that maximum latewood BI (LWBI) chronologies exhibit positive and temporally stable correlations (r = 0.28–0.54, p

Summary: It is well established that the basolateral amygdala (BLA) is an emotional processing hub that governs a diverse repertoire of behaviors. Selective engagement of a heterogeneous cell population in the BLA is thought to contribute to this flexibility in behavioral outcomes. However, whether this process is impacted by previous experiences that influence emotional processing remains unclear. Here we demonstrate that previous positive (enriched environment [EE]) or negative (chronic unpredictable stress [CUS]) experiences differentially influence the activity of populations of BLA principal neurons projecting to either the nucleus accumbens core or bed nucleus of the stria terminalis. Chemogenetic manipulation of these projection-specific neurons can mimic or occlude the effects of CUS and EE on behavioral outcomes to bidirectionally control avoidance behaviors and stress-induced helplessness. These data demonstrate that previous experiences influence the responsiveness of projection-specific BLA principal neurons, biasing information routing through the BLA, to drive divergent behavioral outcomes.

Abstract The southeastern United States Coastal Plain ecosystem contains baldcypress (Taxodium distichum) swamps and grass‐dominated marshes. These ecosystems also occurred on the exposed continental shelf during lower sea levels but are rarely preserved due to the mechanically erosive nature of transgression and regression. Two presently marine sites on the northeastern Gulf of Mexico's continental shelf contain well‐preserved woody terrestrial sediments that were the subject of previous studies. This study continues the investigation using geochemical (δ13C, δ15N, δ34S) and palynological characteristics of these formerly terrestrial sediments to determine if swamps and/or marshes existed at the time of deposition. The first site is located ∼20 km southeast of Horn Island, Mississippi (MS) and the core has terrestrial sediments radiocarbon dated to 11,066–10,228 (2σ) calibrated years BP (early Holocene). The second site is the “Alabama Underwater Forest” located ∼13 km south of Gulf Shores, Alabama (AL) and the cores have terrestrial sediments optically stimulated luminescence dated to 63 ka (±10 ka, 2σ) to 72 ka (±16 ka, 2σ) (late Pleistocene). Geochemical results for the MS sediments indicate a swamp‐to‐freshwater marsh transitional series, whereas the AL sediments indicate a swamp‐to‐saltwater marsh transitional series, both supported by palynological results. Further exploration of the geochemical results using linear discriminant analysis, trained with published geochemical data, supports the swamp and marsh interpretations. We conclude that the near‐pristine preservation of these woody deposits is not solely due to physical mechanisms, such as rapid burial, but is also coupled with anoxia‐ and euxinia‐driven biogeochemical reactions promoting wood and woody debris preservation in swamp and marsh environments.

Abstract Background The mosquito microbiome is an important modulator of vector competence and vectoral capacity. Unlike the extensively studied bacterial microbiome, fungal communities in the mosquito microbiome (the mycobiome) remain largely unexplored. To work towards getting an improved understanding of the fungi associated with mosquitoes, we sequenced the mycobiome of three field-collected and laboratory-reared mosquito species (Aedes albopictus, Aedes aegypti, and Culex quinquefasciatus). Results Our analysis showed both environment and host species were contributing to the diversity of the fungal microbiome of mosquitoes. When comparing species, Ae. albopictus possessed a higher number of diverse fungal taxa than Cx. quinquefasciatus, while strikingly less than 1% of reads from Ae. aegypti samples were fungal. Fungal reads from Ae. aegypti were

Glial fibrillary acidic protein (GFAP) has become the most promising biomarker for detecting traumatic abnormalities on head computed tomography (CT) in patients with traumatic brain injury (TBI), but most studies have not addressed the potential added value of combining the biomarker with clinical variables that confer risk for intracranial injuries. The Scandinavian Guidelines for Initial Management of Minimal, Mild, and Moderate Head Injuries in Adults were the first clinical decision rules in the field with an incorporated biomarker, the S100 astroglial calcium-binding protein B (S100B), which is used in the Mild (Low Risk) group defined by the guidelines. Our aim was to evaluate the performance of the guidelines when S100B was substituted with GFAP. The sample (N?=?296) was recruited from the Tampere University Hospital's emergency department between November 2015 and November 2016, and there were 49 patients with available GFAP results who were stratified in the Mild (Low Risk) group (thus patients undergoing biomarker triaging). A previously reported cutoff of plasma GFAP ?140?pg/mL was used. Within the Mild (Low Risk) group (n?=?49), GFAP sensitivity (with 95% confidence intervals in parentheses) for detecting traumatic CT abnormalities was 1.0 (0.40?1.00), specificity 0.34 (0.19?0.53), the negative predictive value (NPV) 1.0 (0.68?1.00), and the positive predictive value (PPV) 0.16 (0.05?0.37). The sensitivity and specificity of the modified guidelines with GFAP, when applied to all imaged patients (n?=?197) in the whole sample, were 0.94 (0.77?0.99) and 0.20 (0.15?0.28), respectively. NPV was 0.94 (0.80?0.99) and PPV 0.18 (0.13?0.25). In the Mild (Low Risk) group, none of the patients with GFAP results below 140?pg/mL had traumatic abnormalities on their head CT. These findings were derived from a small patient subgroup. Future researchers should replicate these findings in larger samples and assess whether GFAP has added or comparable value to S100B in acute TBI management.

Anthropogenic chemicals are fundamental for maintaining our standard of living in modern society. Unfortunately, some chemicals are persistent and can enter waste streams and, ultimately, the environment. Commonly used household products, including pharmaceuticals and personal care products, are notable sources of contaminants. The aim of this study was to develop an ‘Up-the-Pipe Solutions’ framework to raise awareness within the wider community about the presence and potential risks of the chemicals found in household products. There is potential to reduce the levels of contaminants in waste streams or substitute them with less harmful alternatives by raising awareness. This framework is based on ‘The Natural Step’ and the concept of essentiality; it recognises the importance of engaging with communities to raise awareness of each individual’s contribution to the problem. Our daily activities and behavioural patterns can have adverse consequences, including the release of persistent contaminants in main waste streams from our kitchens, bathrooms, and green wastes. The ‘Up-the-Pipe Solutions’ framework was successfully used in a small community to engage with school children, local authorities, and Māori leaders to raise awareness of chemical pollution.

Objective The primary goal of this study is to evaluate the relationship between Body Mass Index (BMI) and muscle atrophy in individuals with rotator cuff tears.Methods This study consists of patients with rotator cuff tears identified by MRI from two independent cohorts, the Rotator Cuff Outcomes Workgroup (ROW) and the Multicenter Orthopaedic Outcomes Network (MOON). Presence of atrophy (yes/no) and severity of atrophy (as an ordinal variable) were assessed on MRI by expert physicians. We used multivariable regression models to evaluate the relationship between BMI and muscle atrophy while adjusting for age and sex in each study, conducted sensitivity analyses for full-thickness tear and combined results using inverse variance-weighted meta-analysis.Results A total of 539 patients (MOON=395, ROW=144) from the combined cohorts had MRI data available on muscle atrophy. Among these patients, 246 (46%) had atrophy of at least one of the muscles of the rotator cuff and 282 (52%) had full-thickness tears. In meta-analysis across both cohorts, each 5 kg/m2 increase in BMI was associated with a 21% (aOR=1.21, 95% CI=1.02, 1.43) increased odds of having muscle atrophy among individuals with any tear size, and 36% (aOR=1.36, 95% CI=1.01–1.81) increased odds among individuals with full-thickness tear.Conclusions Higher BMI was associated with significantly higher odds of muscle atrophy in patiens with rotator cuff tears. More study is needed to unders1tand why and how this relationship exists, as well as whether interventions to reduce BMI may help improve outcomes for these patients.Level of Evidence III.

ObjectiveAdverse childhood experiences (ACEs) are associated with mental health and cognitive problems, and mental health problems are associated with perceived cognitive difficulties among adolescents. The unique contribution of ACEs to cognitive difficulties after adjusting for poor mental health is not well understood and represents the purpose of this study.MethodsThe Adolescent Behaviors and Experiences Survey was conducted in 2021 with high school students in the United States. Cognitive difficulty was assessed with: ‘Because of a physical, mental, or emotional problem, do you have serious difficulty concentrating, remembering, or making decisions?’ Four ACEs were examined: sexual violence (lifetime and past 12 months), parental emotional abuse, and parental physical abuse. Students were asked about feeling sad or hopeless (past year), considering suicide (past year), and having poor mental health (past month). Binary logistic regressions examined the association between ACEs and cognitive problems, adjusting for mental health.ResultsParticipants were 6,945 students. Students reporting poor mental health were very likely to endorse difficulty concentrating, remembering, or making decisions (girls = 81% and boys = 67%). Cognitive difficulty was uncommon among students who denied poor mental health (girls = 17% and boys = 12%). For boys [p

Background Culex (Cx.) tritaeniorhynchus is an invasive mosquito species with an extensive and expanding inter-continental distribution, currently reported across Asia, Africa, the Middle East, Europe and now Australia. It is an important vector of medical and veterinary pathogens which cause significant morbidity and mortality in human and animal populations. Across regions endemic for Japanese encephalitis virus (JEV), Cx. tritaeniorhynchus is considered the major vector and has also been shown to contribute to the transmission of several other zoonotic arboviruses including Rift Valley fever virus (RVFV) and West Nile virus (WNV). Methods In this study, we used laboratory vector competence experiments to determine if Cx. tritaeniorhynchus from a Southern European population were competent JEV vectors. We also obtained samples from multiple geographically dispersed Cx. tritaeniorhynchus populations from countries within Europe, Africa, Eurasia and Asia to perform phylogenetic analysis to measure the level of mitochondrial divergence using the cytochrome oxidase subunit 1 (CO1) gene. We also undertook bacterial 16S rRNA gene amplicon sequencing to determine microbial diversity and used multi-locus sequence typing (MLST) to determine any evidence for the presence of strains of the naturally occurring endosymbiotic bacterium Wolbachia. Results Cx. tritaeniorhynchus from a Greek population were shown be be competent vectors of JEV with high levels of virus present in saliva. We found a signficant level of mitochondrial genetic diversity using the mosquito CO1 gene between geographically dispersed populations. Furthermore, we report diverse microbiomes identified by 16S rRNA gene amplicon sequencing within and between geographical populations. Evidence for the detection of the endosymbiotic bacteria Wolbachia was confirmed using Wolbachia-specific PCR and MLST. Conclusions This study enhances our understanding of the diversity of Cx. tritaeniorhynchus and the associated microbiome across its inter-continental range and highlights the need for greater surveillance of this invasive vector species in Europe.

Abstract Extreme summer temperatures are increasingly common across the Northern Hemisphere and inflict severe socioeconomic and biological consequences. In summer 2021, the Pacific Northwest region of North America (PNW) experienced a 2-week-long extreme heatwave, which contributed to record-breaking summer temperatures. Here, we use tree-ring records to show that summer temperatures in 2021, as well as the rate of summertime warming during the last several decades, are unprecedented within the context of the last millennium for the PNW. In the absence of committed efforts to curtail anthropogenic emissions below intermediate levels (SSP2–4.5), climate model projections indicate a rapidly increasing risk of the PNW regularly experiencing 2021-like extreme summer temperatures, with a 50% chance of yearly occurrence by 2050. The 2021 summer temperatures experienced across the PNW provide a benchmark and impetus for communities in historically temperate climates to account for extreme heat-related impacts in climate change adaptation strategies.

COVID-19 is a spectrum of clinical symptoms in humans caused by infection with SARS-CoV-2. The coalescence of SARS-CoV-2 with seasonal respiratory viruses, particularly influenza viruses, is a global health concern. To understand this, transgenic mice expressing the human ACE2 receptor (K18-hACE2) were infected with influenza A virus (IAV) followed by SARS-CoV-2 and the host response and effect on virus biology was compared to K18-hACE2 mice infected with IAV or SARS-CoV-2 alone. The sequentially infected mice showed reduced SARS-CoV-2 RNA synthesis, yet exhibited more rapid weight loss, more severe lung damage and a prolongation of the innate response compared to the singly infected or control mice. Sequential infection also exacerbated the extrapulmonary encephalitic manifestations associated with SARS-CoV-2 infection. Conversely, prior infection with a commercially available, multivalent live-attenuated influenza vaccine (Fluenz Tetra) elicited the same reduction in SARS-CoV-2 RNA synthesis, albeit without the associated increase in disease severity. This suggests that the innate immune response stimulated by IAV inhibits SARS-CoV-2. Interestingly, infection with an attenuated, apathogenic influenza vaccine does not result in an aberrant immune response and enhanced disease severity. Taken together, the data suggest coinfection (‘twinfection’) is deleterious and mitigation steps should be instituted as part of the comprehensive public health and management strategy of COVID-19.

MHC restriction depicts CD4+ T cell activation by MHC-II and CD8A+ by MHC-I, but rare T cells with mismatched MHC restriction have been reported. Here the authors use crystallography to describe features of such a mismatched TCR-MHC interaction and implicate an atypical TCR structure and TCR-MHC interface as contributing factors.

Abstract Effective mitigation of the impacts of invasive ship rats (Rattus rattus) requires a good understanding of their ecology, but this knowledge is very sparse for urban and peri-urban areas. We radiomarked ship rats in Wellington, New Zealand, to estimate detection parameters (σ, ε 0 , θ, and g 0 ) that describe the process of an animal encountering a device (bait stations, chew cards and WaxTags) from a distance, and then approaching it and deciding whether to interact with it. We used this information in simulation models to estimate optimal device spacing for eradicating ship rats from Wellington, and for confirming eradication. Mean σ was 25.37 m (SD = 11.63), which equates to a circular home range of 1.21 ha. The mean nightly probability of an individual encountering a device at its home range center (ε 0 ) was 0.38 (SD = 0.11), whereas the probability of interacting with the encountered device (θ) was 0.34 (SD = 0.12). The derived mean nightly probability of an individual interacting with a device at its home range center (g 0 ) was 0.13 (SD = 0.08). Importantly, σ and g 0 are intrinsically linked through a negative relationship, thus g 0 should be derived from σ using a predictive model including individual variability. Simulations using this approach showed that bait stations deployed for about 500 days using a 25 m × 25 m grid consistently achieved eradication, and that a surveillance network of 3.25 chew cards ha−1 or 3.75 WaxTags ha−1 active for 14 nights would be required to confidently declare eradication. This density could be halved if the surveillance network was deployed for 28 nights or if the prior confidence in eradication was high (0.85). These recommendations take no account of differences in detection parameters between habitats. Therefore, if surveillance suggests that individuals are not encountering devices in certain habitats, device density should be adaptively revised. This approach applies to initiatives globally that aim to optimise eradication with limited funding.

The composition of the microbiome is shaped by both environment and host in most organisms, but in the mosquito Aedes aegypti the role of the host in shaping the microbiome is poorly understood. Previously, we had shown that four lines of Ae. aegypti harbored different microbiomes when reared in the same insectary under identical conditions. To determine whether these lines differed from each other across time and in different environments, we characterized the microbiome of the same four lines of Ae. aegypti reared in the original insectary and at another institution. While it was clear that the environment influenced the microbiomes of these lines, we did still observe distinct differences in the microbiome between lines within each insectary. Clear differences were observed in alpha diversity, beta diversity, and abundance of specific bacterial taxa. To determine if the line specific differences in the microbiome were maintained across environments, pair-wise differential abundances of taxa was compared between insectaries. Lines were most similar to other lines from the same insectary than to the same line reared in a different insectary. Additionally, relatively few differentially abundant taxa identified between pairs of lines were shared across insectaries, indicating that line specific properties of the microbiome are not conserved across environments, or that there were distinct microbiota within each insectary. Overall, these results demonstrate that mosquito lines under the same environmental conditions have different microbiomes across microbially- diverse environments and host by microbe interactions affecting microbiome composition and abundance is dependent on environmentally available bacteria.

Abstract Many of the world's native fauna suffer unsustainable losses from invasive mammalian predators. Conservation managers control predators on the premise that if large numbers are removed, prey will respond. This is sometimes true, but not always. Empirical relationships between predator densities and responses of vulnerable prey in Oceania often show little or no response across a broad range of predator reductions, with positive responses only at low threshold predator densities. Even then, some prey populations fail to respond. More research is required to identify predator thresholds across a range of prey taxa. This uncertainty of outcomes, coupled with the considerable cost of mammalian pest control, risks little or no return from limited conservation funds. A unifying theory is required to help understand why conservation outcomes from predator control are so variable despite the best efforts of conservation managers, and to expedite the right kind of management for a given prey species. We argue that a modern synthesis of numerical and functional response theory, in the form of total response models, provides such a theory. Stochastic consumer‐resource models are recommended for dynamic systems, but they are difficult to parameterize. Total response models, on the other hand, present a simple conceptual framework that managers can use as a heuristic to understand predator–prey systems, help explain some of the variability in predator control outcomes and stimulate thinking about other management options that can be integrated with predator control to improve conservation outcomes. Five rules of thumb are suggested to assist conservation managers.

Microplastics and their associated plastic additives are contaminants of emerging environmental concern. The extent and rate of additive leaching affects the behaviour and fate of these contaminants, and therefore the risk they present to organisms. However, most studies examining the leaching of additives from microplastics use methods that are not environmentally relevant. We evaluated additive leaching from a range of common plastics using an environmentally relevant dynamic leaching method with additives and non-intentionally added substances quantified by liquid chromatography-mass spectrometry. The leaching method was validated to ensure sink conditions were maintained over the duration of the test. Release was not solubility limited indicating that the test conditions were representative of those occurring in the environment. Leaching profiles showed significantly different kinetics across the different types of plastic, with cumulative additive release over 64 days spanning more than five orders of magnitude. This study highlights the wide variation in additive leaching behaviour between different plastics and the importance of understanding leaching kinetics to assess the chemical impacts of microplastic pollution. Furthermore, we identify the need for environmentally relevant leaching methods and suggest the creation of standardised methods to enable robust risk assessments and regulation.

IntroductionSome ultra-high exposure boxers from the 20th century suffered from neurological problems characterized by slurred speech, personality changes (e.g., childishness or aggressiveness), and frank gait and coordination problems, with some noted to have progressive Parkinsonian-like signs. Varying degrees of cognitive impairment were also described, with some experiencing moderate to severe dementia. The onset of the neurological problems often began while they were young men and still actively fighting. More recently, traumatic encephalopathy syndrome (TES) has been proposed to be present in athletes who have a history of contact (e.g., soccer) and collision sport participation (e.g., American-style football). The characterization of TES has incorporated a much broader description than the neurological problems described in boxers from the 20th century. Some have considered TES to include depression, suicidality, anxiety, and substance abuse.PurposeWe carefully re-examined the published clinical literature of boxing cases from the 20th century to determine whether there is evidence to support conceptualizing psychiatric problems as being diagnostic clinical features of TES.MethodsWe reviewed clinical descriptions from 155 current and former boxers described in 21 articles published between 1928 and 1999.ResultsMore than one third of cases (34.8%) had a psychiatric, neuropsychiatric, or neurobehavioral problem described in their case histories. However, only 6.5% of the cases were described as primarily psychiatric or neuropsychiatric in nature. The percentages documented as having specific psychiatric problems were as follows: depression = 11.0%, suicidality = 0.6%, anxiety = 3.9%, anger control problems = 20.0%, paranoia/suspiciousness = 11.6%, and personality change = 25.2%.DiscussionWe conclude that depression, suicidality (i.e., suicidal ideation, intent, or planning), and anxiety were not considered to be clinical features of TES during the 20th century. The present review supports the decision of the consensus group to remove mood and anxiety disorders, and suicidality, from the new 2021 consensus core diagnostic criteria for TES. More research is needed to determine if anger dyscontrol is a core feature of TES with a clear clinicopathological association. The present findings, combined with a recently published large clinicopathological association study, suggest that mood and anxiety disorders are not characteristic of TES and they are not associated with chronic traumatic encephalopathy neuropathologic change.

Tropical cyclone precipitation on the US Gulf Coast is greatly influenced by the behavior of the North Atlantic subtropical high and volcanic eruptions as well as by Atlantic Multidecadal Variability, suggests a 473-year-long tree-ring proxy record.

Background: The growing role of biologic therapies as adjunct or standalone procedures in orthopedic practice has led to greater levels of direct-to-consumer biologic marketing. The present study aims to assess the quality, accuracy, and readability of online educational resources available to patients regarding biologic therapies for shoulder pathology. Methods: Eight search terms relevant to shoulder biologic therapies (shoulder + BMAC, Bone Marrow Aspirate Concentrate, PRP, Platelet Rich Plasma, Lipogems, Adipose Tissue, Biologic therapy, and Stem cell therapy) were searched across three separate search engines. The first 25 websites of each search were recorded. Duplicate websites and those not specific to shoulder pathology were excluded. Three evaluators independently assessed quality using an author-derived scoring rubric for a total of 25 possible points and accuracy for a total of 12 possible points. The Flesch-Kincaid readability test was used to quantify reading levels. Websites were further characterized by authorship and the presence of commercial bias. Results: Of the 600 results from the initial search, 59 met inclusion criteria. The mean quality of the websites was poor, with 7.97 ± 2.3 of 25 points (32%). The mean accuracy was low, with 8.47 ± 1.52 of 12 points (71%). The average reading level was 11.2 ± 1.93, with 32% of websites' reading at greater than 12th grade reading level. The search terms of “shoulder PRP” and “shoulder Platelet Rich Plasma” yielded the highest quality results (mean = 8.14 ± 2.63). “shoulder Lipogems” and “shoulder Adipose tissue” yielded the most accurate results (mean = 9.25 ± 0.96). “shoulder BMAC” and “shoulder bone marrow aspirate concentrate” were most difficult to read (mean = 12.54 ± 3.73). Sixty-four percent of websites were authored by physicians, hospitals, or medical groups. The accuracy of websites authored by health care professionals was significantly higher than the accuracy of those authored by other industry sources (P = .01). Fifteen percent of websites demonstrated commercial bias. Discussion: The online resources available to patients seeking information about biologic therapies for the treatment of shoulder pathologies are of very poor quality, moderately poor accuracy, and advanced readability. Providers should caution patients about the reliability of direct-to-consumer biologic marketing for shoulder pathology. Conclusion: The information available to patients online regarding the diagnosis, evaluation, and treatment of shoulder pathology with biologic therapies is of poor quality and accuracy and difficult readability.

Abstract Background Mosquitoes harbor microbial communities that play important roles in their growth, survival, reproduction, and ability to transmit human pathogens. Microbiome transplantation approaches are often used to study host-microbe interactions and identify microbial taxa and assemblages associated with health or disease. However, no such approaches have been developed to manipulate the microbiota of mosquitoes. Results Here, we developed an approach to transfer entire microbial communities between mosquito cohorts. We undertook transfers between (Culex quinquefasciatus to Aedes aegypti) and within (Ae. aegypti to Ae. aegypti) species to validate the approach and determine the number of mosquitoes required to prepare donor microbiota. After the transfer, we monitored mosquito development and microbiota dynamics throughout the life cycle. Typical holometabolous lifestyle-related microbiota structures were observed, with higher dynamics of microbial structures in larval stages, including the larval water, and less diversity in adults. Microbiota diversity in recipient adults was also more similar to the microbiota diversity in donor adults. Conclusions This study provides the first evidence for successful microbiome transplantation in mosquitoes. Our results highlight the value of such methods for studying mosquito-microbe interactions and lay the foundation for future studies to elucidate the factors underlying microbiota acquisition, assembly, and function in mosquitoes under controlled conditions. Video Abstract

ObjectiveThe tackle is the most injurious event in rugby league and carries the greatest risk of concussion. This study aims to replicate previous research conducted in professional men's rugby league by examining the association between selected tackle characteristics and head impact events (HIEs) in women's professional rugby league.MethodsWe reviewed and coded 83 tackles resulting in an HIE and every tackle (6,318 tackles) that did not result in an HIE for three seasons (2018–2020) of the National Rugby League Women's (NRLW) competition. Tackle height, body position of the tackler and ball carrier, as well as the location of head contact with the other player's body were evaluated. Propensity of each situation that caused an HIE was calculated as HIEs per 1,000 tackles.ResultsThe propensity for tacklers to sustain an HIE was 6.60 per 1,000 tackles (95% CI: 4.87–8.92), similar to that of the ball carrier (6.13 per 1,000 tackles, 95% CI: 4.48–8.38). The greatest risk of an HIE to either the tackler or ball carrier occurred when head proximity was above the sternum (21.66 per 1,000 tackles, 95% CI: 16.55–28.35). HIEs were most common following impacts between two heads (287.23 HIEs per 1,000 tackles, 95% CI: 196.98–418.84). The lowest propensity for both tackler (2.65 per 1,000 tackles, 95% CI: 0.85–8.20) and ball carrier HIEs (1.77 per 1,000 tackles, 95% CI: 0.44–7.06) occurred when the head was in proximity to the opponent's shoulder and arm. No body position (upright, bent or unbalanced/off feet) was associated with an increased propensity of HIE to either tackler or ball carrier.ConclusionsIn the NRLW competition, tacklers and ball carriers have a similar risk of sustaining an HIE during a tackle, differing from men's NRL players, where tacklers have a higher risk of HIEs. Further studies involving larger samples need to validate these findings. However, our results indicate that injury prevention initiatives in women's rugby league should focus on how the ball carrier engages in contact during the tackle as well as how the tackler executes the tackle.

IntroductionThis systematic review examined whether race or ethnicity are associated with clinical outcomes (e.g., time to return to school/sports, symptom duration, vestibular deficits, and neurocognitive functioning) following sport-related concussion among child, adolescent, or college-aged student athletes. Additionally, this review assessed whether the existing literature on this topic incorporated or included broader coverage of social determinants of health.MethodsThe online databases PubMed, MEDLINE®, PsycINFO®, CINAHL, Cochrane Library, EMBASE, SPORTDiscus, Scopus, and Web of Science were searched.ResultsA total of 5,118 abstracts were screened and 12 studies met inclusion criteria, including 2,887 youth and young adults. Among the included articles, only 3 studies (25%) examined whether race and ethnicity were associated with outcomes following concussion as a primary objective. None of the studies assessed the association between social determinants of health and outcomes following concussion as a primary objective, although 5 studies (41.7%) addressed a social determinant of health or closely related topic as a secondary objective.DiscussionOverall, the literature to date is extremely limited and insufficient for drawing conclusions about whether race or ethnicity are categorically associated with outcomes from sport-related concussion, or more specifically, whether there are socioeconomic, structural, or cultural differences or disparities that might be associated with clinical outcome.Systematic review registrationidentifier: PROSPERO, CRD42016041479, CRD42019128300.

IntroductionWe examined postmortem brain tissue from men, over the age of 50, for chronic traumatic encephalopathy neuropathologic change (CTE-NC). We hypothesized that (i) a small percentage would have CTE-NC, (ii) those who played American football during their youth would be more likely to have CTE-NC than those who did not play contact or collision sports, and (iii) there would be no association between CTE-NC and suicide as a manner of death.MethodsBrain tissue from 186 men and accompanying clinical information were obtained from the Lieber Institute for Brain Development. Manner of death was determined by a board-certified forensic pathologist. Information was obtained from next of kin telephone interviews, including medical, social, demographic, family, and psychiatric history. The 2016 and 2021 consensus definitions were used for CTE-NC. Two authors screened all cases, using liberal criteria for identifying “possible” CTE-NC, and five authors examined the 15 selected cases.ResultsThe median age at the time of death was 65 years (interquartile range = 57–75; range = 50–96). There were 25.8% with a history of playing American football and 36.0% who had suicide as their manner of death. No case was rated as definitively having “features” of CTE-NC by all five authors. Ten cases were rated as having features of CTE-NC by three or more authors (5.4% of the sample), including 8.3% of those with a personal history of playing American football and 3.9% of those who did not play contact or collision sports. Of those with mood disorders during life, 5.5% had features of CTE-NC compared to 6.0% of those who did not have a reported mood disorder. Of those with suicide as a manner of death, 6.0% had features of CTE-NC compared to 5.0% of those who did not have suicide as a manner of death.DiscussionWe did not identify a single definitive case of CTE-NC, from the perspective of all raters, and only 5.4% of cases were identified as having possible features of CTE-NC by some raters. CTE-NC was very uncommon in men who played amateur American football, those with mood disorders during life, and those with suicide as a manner of death.

Background: Ongoing research of the mosquito microbiome aims to uncover novel strategies to reduce pathogen transmission. Sequencing costs, especially for metagenomics, are however still significant. A resource that is increasingly used to gain insights into host-associated microbiomes is the large amount of publicly available genomic data based on whole organisms like mosquitoes, which includes sequencing reads of the host-associated microbes and provides the opportunity to gain additional value from these initially host-focused sequencing projects. Methods: To analyse non-host reads from existing genomic data, we developed a snakemake workflow called MINUUR (Microbial INsights Using Unmapped Reads). Within MINUUR, reads derived from the host-associated microbiome were extracted and characterised using taxonomic classifications and metagenome assembly followed by binning and quality assessment. We applied this pipeline to five publicly available Aedes aegypti genomic datasets, consisting of 62 samples with a broad range of sequencing depths. Results: We demonstrate that MINUUR recovers previously identified phyla and genera and is able to extract bacterial metagenome assembled genomes (MAGs) associated to the microbiome. Of these MAGS, 42 are high-quality representatives with >90% completeness and

Abstract Climate policies will affect future surface ozone pollution in China. Here, we simulate changes in summertime ozone across China by 2030 under four emission scenarios reflecting different levels of climate action. We also contrast results obtained with two different chemical mechanisms employed in the chemical transport model (WRF‐Chem). With emission reductions in ozone precursors introduced by climate policies, both mechanisms show promising ozone mitigation for most parts of China. However, they disagree starkly in China's three main city clusters, where one mechanism projects worsening ozone pollution by 2030 despite the emission reductions. We analyze possible drivers of this important discrepancy, in particular the role of varying ozone chemical regimes affecting its sensitivity to emission changes. We recommend an intercomparison project to examine this critical modeling uncertainty among other models/mechanisms, which would be invaluable for informing local and regional emission control strategies that are based on single‐model results.

Mosquitoes develop in a wide range of aquatic habitats containing highly diverse and variable bacterial communities that shape both larval and adult traits, including the capacity of adult females of some mosquito species to transmit disease-causing organisms to humans. However, while most mosquito studies control for host genotype and environmental conditions, the impact of microbiota variation on phenotypic outcomes of mosquitoes is often unaccounted for. The inability to conduct reproducible intra- and inter-laboratory studies of mosquito-microbiota interactions has also greatly limited our ability to identify microbial targets for mosquito-borne disease control. Here, we developed an approach to isolate and cryopreserve bacterial communities derived from lab and field-based larval rearing environments of the yellow fever mosquito Aedes aegypti-a primary vector of dengue, Zika, and chikungunya viruses. We then validated the use of our approach to generate experimental microcosms colonized by standardized lab- and field-derived bacterial communities. Our results overall reveal minimal effects of cryopreservation on the recovery of both lab- and field-derived bacteria when directly compared with isolation from non-cryopreserved fresh material. Our results also reveal improved reproducibility of bacterial communities in replicate microcosms generated using cryopreserved stocks over fresh material. Communities in replicate microcosms further captured the majority of total bacterial diversity present in both lab- and field-based larval environments, although the relative richness of recovered taxa as compared to non-recovered taxa was substantially lower in microcosms containing field-derived bacteria. Altogether, these results provide a critical next step toward the standardization of mosquito studies to include larval rearing environments colonized by defined microbial communities. They also lay the foundation for long-term studies of mosquito-microbe interactions and the identification and manipulation of taxa with potential to reduce mosquito vectorial capacity.

Abstract Severe acute respiratory coronavirus 2 (SARS-CoV-2) has spread globally since its emergence in 2019. Most SARS-CoV-2 infections generate immune responses leading to rising levels of immunoglobulins (Ig) M, A and G which can be detected using diagnostic tests including enzyme-linked immunosorbent assays (ELISA). Whilst implying previous SARS-CoV-2 infection, the detection of Ig by ELISA does not guarantee the presence of neutralising antibodies (NAb) that can prevent the virus infecting cells. Plaque reduction neutralisation tests (PRNT) detect NAb, but are not amenable to mass testing as they take several days and require use of SARS-CoV-2 in high biocontainment laboratories. We evaluated the ability of IgG and IgM ELISAs targeting SARS-CoV-2 spike subunit 1 receptor binding domain (S1-RBD), and spike subunit 2 (S2) and nucleocapsid protein (NP), at predicting the presence and magnitude of NAb determined by PRNT. IgG S2 + NP ELISA was 96.8% [95% CI 83.8–99.9] sensitive and 88.9% [95% CI 51.8–99.7] specific at predicting the presence of NAbs (PRNT80 > 1:40). IgG and IgM S1-RBD ELISAs correlated with PRNT titre, with higher ELISA results increasing the likelihood of a robust neutralising response. The IgM S1-RBD assay can be used as a rapid, high throughput test to approximate the magnitude of NAb titre.

The authors study mechanisms underlying neuromodulatory control of transitions between brain and behavioral states. They identify a mechanism whereby modulation of Gq activity in basolateral amygdala parvalbumin interneurons mediates the transition to a fear-associated network and behavioral state.

Microplastics and plastic additives are contaminants of emerging environmental concern. Static leaching methods are commonly applied to assess the rate and extent of additive release from microplastics. However, this approach may not be representative of environmental conditions where near infinite dilution or percolation commonly occur. We evaluated three different approaches for assessing additive leaching under environmentally relevant sink conditions, culminating in the refinement and validation of DyLeMMA (Dynamic Leaching Method for Microplastic Assessment). Analysis was performed using a high-resolution liquid chromatography-mass spectrometry method enabling targeted quantification of additives and screening for non-intentionally added substances. Using four different plastics, sink conditions were maintained over the duration of the test, thereby avoiding solubility limited release and ensuring environmental relevance. Background contamination from ubiquitous additive chemicals was minimised, thereby providing good sensitivity and specificity. Resulting data, in the form of additive release curves, should prove suitable for fitting to release models and derivation of parameters describing additive leaching from microplastics.Key attributes of DyLeMMA: • Environmentally relevant dynamic leaching method for microplastics, demonstrated to maintain sink conditions over the test duration, • Simple, fast, and cost-effective approach without complication of using a solid phase sink, • Provide data suitable for understanding microplastic leaching kinetics and mechanisms.

Knockdown resistance (kdr) alleles conferring resistance to pyrethroid insecticides are widespread amongst vector populations. Previous research has suggested that these alleles are associated with changes in the vector competence of mosquitoes for arboviruses and Plasmodium, however non-target genetic differences between mosquito strains may have had a confounding effect. Here, to minimise genetic differences, the laboratory Anopheles gambiae Kisumu strain was compared to a CRISPR/Cas9 homozygous kdr L1014F mutant Kisumu-kdr line in order to examine associations with vector competence for o'nyong nyong virus (ONNV). Mosquitoes were infected using either blood feeds or intrathoracic microinjections. There were no significant differences in the prevalence of virus in mosquito body parts between kdr mutant and wildtype lines from either oral or intrathoracic injection routes. The ONNV titre was significantly higher in the legs of the wildtype strain at 7dpi following intrathoracic microinjection, but no other significant differences in viral titre were detected. ONNV was not detected in the saliva of mosquitoes from either strain. Our findings from per os infections suggest that the kdr L1014F allele is not associated with altered infection prevalence for ONNV, a key component of vector competence.

Some people experience persistent post-concussion symptoms (PPCS) after mild traumatic brain injury (mTBI). A meaningful clinical classification and scientific progress are hampered by a lack of consensus regarding the phenomenology, assessment, and operationalization of PPCS. Here we demonstrate and evaluate how the methodology used to assess and define persistent symptoms after mTBI influences PPCS as a binary outcome. We present empirical data from 15 classification methods reflecting procedures found in the literature and clinical practice. In total, 221 patients with mTBI, 73 patients with orthopedic injuries, and 77 community controls were included in the study. The prevalence rate of PPCS in the mTBI group varied between 10% and 47%, depending on the method used to assess and define unfavorable outcome. There was generally low positive agreement between the different methods; even the two methods yielding the most similar prevalence rates (89.2% overall proportion agreement) agreed on less than half (45.5% positive agreement) of the PPCS cases. Using a liberal but not uncommon threshold for symptom severity, there was a considerable misclassification rate of PPCS in both comparison groups. Our results highlight the importance for researchers to be aware of the limitations of using binary approaches for classification of PPCS. The poor agreement between methods should be considered when (1) interpreting the heterogeneity in the existing PPCS literature and (2) developing new improved methods. An empirically informed consensus regarding classification of PPCS should be a priority for the research community.

The immunogenicity of peptides is believed to be determined by their high-affinity binding to MHC I. Here authors show that low-affinity MHC I-peptide interactions are also able to trigger robust T cell response and anti-tumour immunity in vivo.

Abstract Background Malaria control relies mainlyon insecticide-based tools. However, the effectiveness of these tools is threatened by widespread insecticide resistance in malaria vectors, highlighting the need for alternative control approaches. The endosymbiont Asaia has emerged as a promising candidate for paratransgenic control of malaria, but its biology and genetics still need to be further analyzed across Africa. Here, we investigated the prevalence of Asaia and its maternal transmission in the natural population of Anopheles mosquitoes in Cameroon. Methods Indoor-resting adult mosquitoes belonging to four species (An. coluzzii, An. arabiensis, An. funestus and An. gambiae) were collected from eight localities across Cameroon from July 2016 to February 2020. PCR was performed on the Asaia-specific 16S ribosomal RNA gene, and samples positive by PCR for Asaia were confirmed by Sanger sequencing and phylogenetic analysis. The vertical transmission of Asaia was investigated by screening F 1 mosquitoes belonging to F 0 Asaia-positive females. Results A total of 895 mosquitoes were screened. We found 43% (384) Asaia infection prevalence in four mosquito species. Phylogenetic analysis revealed that Asaia from Cameroon clustered together with the strains of Asaia isolated from other parts of the world. In addition, seven nucleotide sequence variants were found with low genetic diversity (π = 0.00241) and nucleotide sequence variant diversity (Hd = 0.481). Asaia was vertically transmitted with high frequency (range from 42.5 to 100%). Conclusions This study provides field-based evidence of the presence of Asaia in Anopheles mosquitoes in Cameroon for exploitation as a symbiont in the control of malaria in sub-Saharan Africa. Graphical abstract

BackgroundSerotonergic antidepressants may predispose to bleeding, but little is known of the risk for traumatic intracranial bleeding.MethodsThis was a prospective case-control study of 218 patients with mild traumatic brain injuries (TBI) who were treated at a Finnish tertiary trauma hospital. Injury-related information and clinical findings were prospectively collected in the emergency department. Detailed pre-injury health history was collected from electronic medical records. Information on the use of serotonergic antidepressants was attained from the Finnish national prescription registry. All head CT scans were reviewed by a neuroradiologist based on the Common Data Elements. Cases were patients with traumatic intracranial hemorrhage on head CT. Controls were patients from the same cohort, but without traumatic intracranial lesions on CT. The proportion with traumatic intracranial bleeding for patients on serotonergic antidepressant medication was compared to the proportion for patients not on serotonergic medication.ResultsThe study cohort consisted of 24 cases with traumatic intracranial bleeding and 194 injured controls. The median age of the sample was 70 years (interquartile range = 50–83). One fifth (21.6%) of all the patients were taking a serotonergic antidepressant. Of the patients on an antidepressant, 10.6% (5/47) had an acute hemorrhagic lesion compared to 11.1% (19/171) of those who were not on an antidepressant (p = 0.927). In the regression analysis, traumatic intracranial hemorrhage was not associated with antidepressant use.ConclusionSerotonergic antidepressant use was not associated with an increased risk of traumatic intracranial hemorrhage after a mild TBI. The patients in this relatively small cohort were mostly middle-aged and older adults. These factors limit the generalizability of the results in younger patients with mild TBI.

Objectives:. Trauma is a leading cause of death worldwide and in South Africa. We aimed to quantify the in-hospital trauma mortality rate in Pietermaritzburg, South Africa. Background:. The in-hospital trauma mortality rate in South Africa remains unknown, and it is unclear whether deficits in hospital care are contributing to the high level of trauma-related mortality. Methods:. All patients hospitalized because of trauma at the Department of Surgery at Grey’s Hospital, Pietermaritzburg Metropolitan Trauma Service, were prospectively entered in an electronic database starting in 2013 and the data were retrospectively analyzed. The trauma service adheres to Advanced Trauma Life Support and the doctors have attended basic and advanced courses in trauma care. The primary outcome was in-hospital mortality. Results:. Of 9795 trauma admissions, 412 (4.2%) patients died during hospital care between January 2013 and January 2019. Forty-six percent died after road traffic accidents, 19% after gunshot wounds, 13% after stab wounds, and 10% after assaults. Sixteen percent were classified as avoidable deaths due to inappropriate care and resource limitations. Fifty percent died because of traumatic brain injury and 80% of them were unavoidable. Conclusions:. In conclusion, the in-hospital trauma mortality rate at a South African trauma center using systematic trauma care is lower than that reported from other trauma centers in the world during the past 20 years. Nevertheless, 16% of death cases were assessed as avoidable if there had been better access to intensive care, dialysis, advanced respiratory care, blood for transfusion, and improvements in surgery and medical care.

ObjectiveImpaired balance and postural stability can occur with advanced age, following traumatic brain injury, in association with neurological disorders and diseases, and as the result of acute or chronic orthopedic problems. The remote assessment of balance and postural stability could be of value in clinical practice and research. We examined the test-retest reliability and reliable change estimates for Sway Balance Mobile Application tests (Sway Medical, Tulsa OK, USA) administered remotely from the participant's home.MethodPrimarily young, healthy community-dwelling adults completed Sway Balance Mobile Application tests remotely on their personal mobile devices once per week for three consecutive weeks while being supervised with a video-based virtual connection. Sway Balance tests include five stances (i.e., feet together, tandem right foot forward, tandem left foot forward, single leg right foot, single leg left foot), which are averaged to compute a Sway Balance composite score from 0 to 100, with higher scores indicating better postural stability. We examined test-retest reliability (measured with intraclass correlation coefficients, ICCs) and preliminary reliable change estimates for 70%, 80%, and 90% confidence intervals.ResultsParticipants included 55 healthy adults (ages = 26.7 ± 9.9 years, interquartile range = 20–30, range = 18–58; 38 [69%] women). Test-retest reliability for the Sway Balance composite score across three weeks was.88. Test-retest reliability for individual stances ranged from 62 to 83 (all ps