Search

Abstract This study investigates the effects of varying loading conditions on excitability in neural pathways and gait dynamics. We focussed on evaluating the magnitude of the Hoffman reflex (H‐reflex), a neurophysiological measure representing the capability to activate motor neurons and the timing and placement of the foot during walking. We hypothesized that weight manipulation would alter H‐reflex magnitude, footfall and lower body kinematics. Twenty healthy participants were recruited and subjected to various weight‐loading conditions. The H‐reflex, evoked by stimulating the tibial nerve, was assessed from the dominant leg during walking. Gait was evaluated under five conditions: body weight, 20% and 40% additional body weight, and 20% and 40% reduced body weight (via a harness). Participants walked barefoot on a treadmill under each condition, and the timing of electrical stimulation was set during the stance phase shortly after the heel strike. Results show that different weight‐loading conditions significantly impact the timing and placement of the foot and gait stability. Weight reduction led to a 25% decrease in double limb support time and an 11% narrowing of step width, while weight addition resulted in an increase of 9% in step width compared to body weight condition. Furthermore, swing time variability was higher for both the extreme weight conditions, while the H‐reflex reduced to about 45% between the extreme conditions. Finally, the H‐reflex showed significant main effects on variability of both stance and swing phases, indicating that muscle‐motor excitability might serve as feedback for enhanced regulation of gait dynamics under challenging conditions.

Abstract Roadside habitat has been touted as a conservation opportunity for insect pollinators, including the declining monarch butterfly. The spectacular monarch migration is under threat from the loss of habitat and the decline of their milkweed host plants. In the northern part of their range, roadsides could potentially produce millions of monarchs annually due to high densities of milkweed; however, roadside milkweed can accumulate chemicals from roads, such as sodium from road salt. Controlled lab studies have shown mixed effects of sodium on monarch development: small increases can be beneficial as sodium is an important micronutrient in brain and muscle development, but large increases can sometimes decrease survival. It is unclear how dietary sodium affects performance in ecologically relevant conditions and the migration itself. In this experiment, we raised monarchs outdoors, in migration‐inducing conditions, on milkweed sprayed with three levels of sodium chloride. We released 2464 tagged monarchs and held an additional 246 for further lab assays. While our recovery rates to the wintering grounds were low (N = 7 individuals), individuals from all three sodium chloride treatments made it to Mexico. Butterflies reared on control milkweed and low salt concentrated sodium in their tissues, while those on high salt diets excreted sodium, suggesting high salt levels were above a physiological optimum. There were no effects of treatment on wing coloration, survival, body size, immunity, or parasite prevalence. Taken together, our results suggest that monarchs are robust to levels of sodium in milkweeds found along roadsides, which is promising with respect to the toxicity of roadside plants.

Abstract Offshore oil and gas platforms support abundant reef fish and are popular fishing sites for recreational anglers. However, the rapid decommissioning and removal of active platforms have decreased such fishing opportunities in the Gulf of Mexico, raising concerns about fisheries impacts. Conversely, planned offshore energy structures like wind turbines may offer similar habitats and fishing sites. To inform spatial planning for marine energy infrastructure in the context of recreational fisheries, we created models of fish communities associated with oil and gas platforms using existing abundance data. We employed Random Forest analysis to predict the presence‐absence and abundance of Red Snapper (Lutjanus campechanus) and Greater Amberjack (Seriola dumerili) at platforms using 47 environmental and platform variables. Nonmetric multidimensional scaling on Bray−Curtis dissimilarities explored fish species composition among 37 species. Results showed variability in Red Snapper and Greater Amberjack incidence/abundance from shore to shelf‐edge, not attributed to surrounding habitat or climatological oceanographic variables. Incidence models were more robust than abundance models. Fish species composition was significantly influenced by location gradient, with less impact from other habitat features. Our findings guide selecting areas for artificial structures to enhance angler opportunities and maintain fish diversity, but identifying the drivers of finer scale abundance variation will require further sampling.

Abstract Monarch butterflies (Danaus plexippus) have undergone large scale population declines leading to multiple conservation organizations encouraging the creation of monarch butterfly habitat. Monarch Watch encourages volunteers to create and register monarch habitat as Monarch Waystations. During registration waystations can be named. Names reflect personal meanings, community attachments, and cultural insights and waystation names in particular can provide insights into what ideas and concepts volunteers associate with a habitat creation program. A total of 37,916 registered waystation names were examined to determine most commonly used names, words and word themes. The most common types of waystations were medium‐sized suburban gardens in the United States. The most common waystation names referenced yard placement with My Backyard the most frequently used. Place‐themed words were the most used followed by animal‐themed words. Within animal‐themed words, insect words were the most used followed by mammals and birds. Theological/mortality themed words were present with memorial the most common word, implying a permanence of waystations. The most common familial terms included mom and grandma suggesting inter‐generational gardening is occurring, which is known to strengthen communities and build well‐being. The frequency of positive emotion/feeling words implies that participating in habitat creation programs is a positive experience and participation may reduce conservation apathy. Further research into habitat creation programs and the longevity of habitat created as well as the motivations and demographics of participants can help tailor habitat volunteer programs to desired audiences and increase overall participation. Read the free Plain Language Summary for this article on the Journal blog.

Abstract Delivering cargo to the central nervous system (CNS) remains a pharmacological challenge. For infectious diseases such as HIV, the CNS acts as a latent reservoir that is inadequately managed by systemic antiretrovirals (ARTs). ARTs thus cannot eradicate HIV, and given CNS infection, patients experience neurological deficits collectively referred to as “neuroHIV”. Herein, the development of bioinspired ionic liquid‐coated nanoparticles (IL‐NPs) for in situ hitchhiking on red blood cells (RBCs) is reported, which enables 48% brain delivery of intracarotid arterial‐ infused cargo. Moreover, IL choline trans‐2‐hexenoate (CA2HA 1:2) demonstrates preferential accumulation in parenchymal microglia over endothelial cells post‐delivery. This study further demonstrates successful loading of abacavir (ABC), an ART that is challenging to encapsulate, into IL‐NPs, and verifies retention of antiviral efficacy in vitro. IL‐NPs are not cytotoxic to primary human peripheral blood mononuclear cells (PBMCs) and the CA2HA 1:2 coating itself confers notable anti‐viremic capacity. In addition, in vitro cell culture assays show markedly increased uptake of IL‐NPs into neural cells compared to bare PLGA nanoparticles. This work debuts bioinspired ionic liquids as promising nanoparticle coatings to assist CNS biodistribution and has the potential to revolutionize the delivery of cargos (i.e., drugs, viral vectors) through compartmental barriers such as the blood‐brain‐barrier (BBB).

Abstract Analysis of environmental DNA (eDNA) has gained widespread usage for taxonomically based biodiversity assessment. While interest in applying noninvasive eDNA monitoring for population genetic assessments has grown, its usage in this sphere remains limited. One barrier to uptake is that the effectiveness of eDNA detection below the species level remains to be determined for multiple species and environments. Here, we test the utility of this emergent technology to obtain within‐species haplotypic variation of New Zealand (NZ) blackfoot pāua (Haliotis iris). We compare mitochondrial haplotype diversity recovered from marine eDNA samples against traditional tissue samples of blackfoot pāua collected at the same NZ coastal site. Targeting the ATP8‐ATP6 region, we recovered four mitochondrial haplotypes from eDNA versus six haplotypes from tissue samples. Three common haplotypes were recovered with both eDNA and tissue samples, while only one out of three rare haplotypes – represented in tissue samples by one individual each – was recovered with our eDNA methods. We demonstrate that eDNA monitoring is an effective tool for recovering common genetic diversity from pāua, although rare (

Abstract Conservation translocations often inherently involve a risk of genetic diversity loss, and thus loss of adaptive potential, but this risk is rarely quantified or monitored through time. The reintroduction of beavers to Scotland, via the Scottish Beaver Trial in Knapdale, is an example of a translocation that took place in the absence of genetic data for the founder individuals and resulted in a small and suspected to be genetically depauperate population. In this study we use a high‐density SNP panel to assess the genetic impact of that initial translocation and the effect of subsequent reinforcement translocations using animals from a different genetic source to the original founders. We demonstrate that the initial translocation did, indeed, lead to low genetic diversity (Ho = 0.052) and high mean kinship (KING‐robust = 0.159) in the Knapdale population compared to other beaver populations. We also show that the reinforcement translocations have succeeded in increasing genetic diversity (Ho = 0.196) and reducing kinship (KING robust = 0.028) in Knapdale. As yet, there is no evidence of admixture between the two genetic lineages that are now present in Knapdale and such admixture is necessary to realise the full genetic benefits of the reinforcement and for genetic reinforcement and then rescue to occur; future genetic monitoring will be required to assess whether this has happened. We note that, should admixture occur, the Knapdale population will harbour combinations of genetic diversity not currently seen elsewhere in Eurasian beavers, posing important considerations for the future management of this population. We consider our results in the wider context of beaver conservation throughout Scotland and the rest of Britain, and advocate for more proactive genetic sampling of all founders to allow the full integration of genetic data into translocation planning in general.

Abstract The lithology, geochemistry, and architecture of the continental lithospheric mantle (CLM) underlying the Kimberley Craton of north‐western Australia has been constrained using pressure‐temperature estimates and mineral compositions for >5,000 newly analyzed and published garnet and chrome (Cr) diopside mantle xenocrysts from 25 kimberlites and lamproites of Mesoproterozoic to Miocene age. Single‐grain Cr diopside paleogeotherms define lithospheric thicknesses of 200–250 km and fall along conductive geotherms corresponding to a surface heat flow of 37–40 mW/m2. Similar geotherms derived from Miocene and Mesoproterozoic intrusions indicate that the lithospheric architecture and thermal state of the CLM has remained stable since at least 1,000 Ma. The chemistry of xenocrysts defines a layered lithosphere with lithological and geochemical domains in the shallow (150 km) CLM, separated by a diopside‐depleted and seismically slow mid‐lithosphere discontinuity (100–150 km). The shallow CLM is comprised of Cr diopsides derived from depleted garnet‐poor and spinel‐bearing lherzolite that has been weakly metasomatized. This layer may represent an early (Meso to Neoarchean?) nucleus of the craton. The deep CLM is comprised of high Cr2O3 garnet lherzolite with lesser harzburgite, and eclogite. The peridotite components are inferred to have formed as residues of polybaric partial mantle melting in the Archean, whereas eclogite likely represents former oceanic crust accreted during Paleoproterozoic subduction. This deep CLM was metasomatized by H2O‐rich melts derived from subducted sediments and high‐temperature FeO‐TiO2 melts from the asthenosphere.

Abstract Many populations of long‐distance migrant shorebirds are declining rapidly. Since the 1970s, the lesser yellowlegs (Tringa flavipes) has experienced a pronounced reduction in abundance of ~63%. The potential causes of the species' decline are complex and interrelated. Understanding the timing of migration, seasonal routes, and important stopover and non‐breeding locations used by this species will aid in directing conservation planning to address potential threats. During 2018–2022, we tracked 118 adult lesser yellowlegs using GPS satellite tags deployed on birds from five breeding and two migratory stopover locations spanning the boreal forest of North America from Alaska to Eastern Canada. Our objectives were to identify migratory routes, quantify migratory connectivity, and describe key stopover and non‐breeding locations. We also evaluated predictors of southbound migratory departure date and migration distance. Individuals tagged in Alaska and Central Canada followed similar southbound migratory routes, stopping to refuel in the Prairie Pothole Region of North America, whereas birds tagged in Eastern Canada completed multi‐day transoceanic flights covering distances of >4000 km across the Atlantic between North and South America. Upon reaching their non‐breeding locations, lesser yellowlegs populations overlapped, resulting in weak migratory connectivity. Sex and population origin were significantly associated with the timing of migratory departure from breeding locations, and body mass at the time of GPS‐tag deployment was the best predictor of southbound migratory distance. Our findings suggest that lesser yellowlegs travel long distances and traverse numerous political boundaries each year, and breeding location likely has the greatest influence on migratory routes and therefore the threats birds experience during migration. Further, the species' dependence on wetlands in agricultural landscapes during migration and the non‐breeding period may make them vulnerable to threats related to agricultural practices, such as pesticide exposure.

Abstract To improve the understanding of the formation and evolution of the sub‐continental lithospheric mantle (SCLM) underlying the South Australian Craton we have conducted a detailed petrological study on >3,000 mantle xenocrysts from 13 kimberlites emplaced across the craton. Pressure (P) and temperature (T) estimates on Cr diopside and garnet have been coupled with their chemical concentrations to constrain lithospheric thickness and chemo‐lithostratigraphy. We show that lithospheric thickness is greatest beneath the Gawler Craton, whereas thinner lithosphere occurs beneath the Adelaide Fold Belt. Mineral compositions highlight two litho‐chemical domains within the shallow and deep SCLM that are separated by a mid‐lithosphere discontinuity (MLD). The shallow SCLM (60–130 km) comprises low Cr2O3 lherzolite and wehrlite. Shallow SCLM xenocrysts record depleted and refertilized compositions enriched in light rare earth elements related to metasomatism by kimberlite or related melts. The mid‐lithosphere (130–160 km) is depleted in garnet and Cr diopside which may relate to a layer of pargasite lherzolite. The deep SCLM (>160 km) comprises high Cr2O3 lherzolite with elevated TiO2 and FeO. We interpret the litho‐chemical stratification of the SCLM to reflect a multi‐stage top‐down growth. The shallow SCLM reflects an amalgamation of Precambrian cratonic nuclei characterized by heterogeneity in geochemical enrichment and depletion. Interaction of the shallow SCLM with mantle plumes accreted melts along the paleo‐lithosphere‐asthenosphere boundary, which now occurs as a MLD. The deep SCLM represents depleted mantle residue formed during mantle plume impingement and thickened during orogenesis. This domain has been metasomatized and refertilized by high‐T melts from the asthenosphere.

Background To investigate oral leukoplakia (OL) and risk of upper gastrointestinal (UGI) cancer deaths in the Linxian Dysplasia Nutrition Intervention Trial (NIT) cohort. Methods A total of 3318 subjects with esophageal squamous dysplasia enrolled on 1 May 1985, and were followed up until 30 September 2015. Participants with OL at baseline were treated as an exposed group, while the remainder was selected as a control group. All subjects were followed monthly and reviewed quarterly by the Linxian Cancer Registry. Cox proportional hazard model was used to estimate hazard ratios (HRs) and 95% confidence intervals (95% CIs). Results During the 30‐year follow‐up, a total of 902 UGI cancer deaths occurred, including 541 esophageal squamous cell carcinoma (ESCC) related, 284 gastric cardia carcinoma (GCC) related, and 77 gastric noncardia carcinoma (GNCC) related deaths. Relative to subjects without OL, the long‐term risk of ESCC mortality in participants with OL increased by 26.1% (HR = 1.26, 95% CI: 1.05–1.52). In the subgroup analyses, adverse effects of OL on ESCC mortality were observed especially in younger subjects (HR = 1.48, 95% CI: 1.11–1.97), females (HR = 1.44, 95% CI: 1.11–1.89), non‐smokers (HR = 1.44, 95% CI: 1.15–1.81), nondrinkers (HR = 1.28, 95% CI: 1.04–1.57), and individuals with a family history of cancer (HR = 1.37, 95% CI: 1.05–1.79). No associations were observed between OL and risk of GCC and GNCC mortality. Conclusions OL may increase the long‐term risk of ESCC mortality, especially in younger subjects, females, nondrinkers, non‐smokers, and subjects with a family cancer history. Future studies are needed to explore the potentially etiological mechanism.

Abstract Background Prolonged cytotoxic concentrations of cytarabine (CA) are required for maximum cytotoxicity. DepoCyt is a human liposomal cytarabine (LC) product that lasts longer in plasma and CSF compared with free CA (FC). The use of LC has not been evaluated in dogs. Objectives To perform a LC pharmacokinetic (PK) study when administered SC in dogs. Animals Five healthy female beagles. Methods Three‐period, 3‐treatment, nonblinded, randomized, and crossover design, including a pilot study. LC was administered at 50 mg/m2 SC and FC was administered at 25 and 50 mg/m2 SC and IV. Plasma CA concentrations were measured until 240, 72, and 8 hours after SC LC, SC FC, and IV FC administration, respectively. CA plasma concentrations were quantitated by ultra‐high‐performance liquid chromatography with mass spectrometry (MS/MS) detection and concentration‐time profiles were evaluated by noncompartmental analysis. Results Subcutaneous LC administration resulted in a maximum plasma concentration of 26.3 to 59.78 ng/mL, time to reach maximum plasma concentration of 2 hours, area under the concentration‐time curve to last measurable concentration of 669.3 to 1126 h × ng/mL, and plasma bioavailability (%F) of 19.6% to 31.3%. The PK profiles of FC after SC and IV administration differed when compared with LC. Conclusions and Clinical Importance In healthy dogs, SC LC administration at 50 mg/m2 results in measurable plasma CA concentrations, is apparently safe and well tolerated, but does not result in prolonged cytotoxic plasma concentrations. Poor absorption of LC prevented establishment of a complete LC PK profile.

Background The objective of this study was to explore the association between fresh fruit consumption and long‐term risk of upper gastrointestinal cancer (UGI) in the Linxian Dysplasia Nutrition Intervention Trial (NIT) cohort. Methods A cohort of 3318 subjects with esophageal squamous dysplasia participated in the Linxian Dysplasia NIT in May 1985 and were followed up until 30 September 2015. Demographic characteristics, lifestyle, and history of diseases were collected at the baseline. The primary endpoint was death from esophageal squamous cell carcinoma (ESCC), gastric cardia carcinoma (GCC), and gastric noncardia carcinoma (GNCC). Hazard ratios (HRs) and 95% confidence intervals (95% CIs) were estimated using the Cox proportional hazard model. Results In the 30‐year follow‐up, a total of 541 ESCC, 284 GCC, and 77 GNCC deaths occurred. Relative to those who never or rarely consumed fresh fruit, the risk of ESCC mortality in participants who consumed fresh fruit more than 12 times/year were significantly decreased by 37.3% (HR = 0.63, 95% CI: 0.49–0.81). In the subgroup analyses, significantly protective effects on ESCC mortality were observed especially in females (HR = 0.59, 95% CI: 0.40–0.89), non‐smokers (HR = 0.67, 95% CI: 0.48–0.94), and nondrinkers (HR = 0.69, 95% CI: 0.51–0.93). Conclusions Consuming fresh fruit more than 12 times/year may reduce the long‐term risk of ESCC mortality in this dysplasia population, particularly in females, non‐smokers, and nondrinkers. Future studies are needed to confirm these findings.

Abstract Commercial fisheries yield essential foods, sustain cultural practices, and provide widespread employment around the globe. Commercially harvested species face a myriad of anthropogenic threats including degraded habitats, changing climate, overharvest, and pollution. Microplastics are pollutants of increasing concern, which are pervasive in the environment and can harbor or adsorb pollutants from surrounding waters. Aquatic organisms, including commercial species, encounter and ingest microplastics, but there is a paucity of data about those caught and cultured in North America. Additional research is needed to determine prevalence, physiological effects, and population‐level implications of microplastics in commercial species from Canada, the United States, and Mexico. Investigations into possible human health effects of microplastic exposure from seafood are also greatly needed. This synthesis summarizes current knowledge, identifies data gaps, and provides future research directions for addressing microplastics effects in commercially valuable North American fishery species.

Abstract Microplastics are of increasing concern as they are readily ingested by aquatic organisms. This study investigated microplastic trophic transfer using larval inland silversides (Menidia beryllina) (5 d posthatch) and unicellular tintinnid (Favella spp.) as a model food chain relevant to North American estuaries. Low‐density polyethylene microspheres (10–20 μm) were used to compare direct ingestion of microplastics by larval fish and trophic transfer via tintinnid prey. Dichlorodiphenyltrichloroethane (DDT)‐treated microspheres were used to determine sorbed pollutant effects on microplastic ingestion. Larval fish exposed directly to microspheres ingested significantly fewer than those exposed via contaminated prey. Larvae ingested significantly more ciliates containing DDT‐treated microspheres than ciliates containing untreated plastics but did not discriminate when exposed directly. Larvae reared for 16 d following a direct 2 h exposure had significantly lower wet weight values than unexposed controls. Our results demonstrate that trophic transfer is a significant route of microplastic exposure that can cause detrimental effects in sensitive life stages.

Abstract Extracellular vesicles (EVs) are of growing interest due to their potential diagnostic, disease surveillance, and therapeutic applications. While several studies have evaluated EV isolation methods in various biofluids, there are few if any data on these techniques when applied to stool. The latter is an ideal biospecimen for studying EVs and colorectal cancer (CRC) because the release of tumour markers by luminal exfoliation into stool occurs earlier than vascular invasion. Since EV release is a conserved mechanism, bacteria in stool contribute to the overall EV population. In this study, we assessed five EV separation methods (ultracentrifugation [UC], precipitation [EQ‐O, EQ‐TC], size exclusion chromatography [SEC], and ultrafiltration [UF]) for total recovery, reproducibility, purity, RNA composition, and protein expression in stool supernatant. CD63, TSG101, and ompA proteins were present in EV fractions from all methods except UC. Human (18s) and bacterial (16s) rRNA was detected in stool EV preparations. Enzymatic treatment prior to extraction is necessary to avoid non‐vesicular RNA contamination. Ultrafiltration had the highest recovery, RNA, and protein yield. After assessing purity further, SEC was the isolation method of choice. These findings serve as the groundwork for future studies that use high throughput omics technologies to investigate the potential of stool‐derived EVs as a source for novel biomarkers for early CRC detection.

Abstract Background The utility of environmental DNA (eDNA) metabarcoding surveys to accurately detect species depends on the degree of DNA dispersal. Multiple marine studies have observed only minimal eDNA transport by horizontal water movement across small spatial scales, leading to the conclusion that spatially specific eDNA signals accurately resemble in‐field species assemblages along a horizontal axis. Marine communities, however, are also structured vertically according to depth. In marine environments displaying permanent water stratification, vertical zonation patterns may be more apparent and present on smaller spatial scales (i.e., meters) than horizontal community structuring. The scale at which eDNA signals differ along a vertical transect and the accuracy of eDNA metabarcoding in revealing naturally stratified communities have yet to be assessed. Methods and results In this study, we determined the ability of eDNA metabarcoding surveys to distinguish vertically localized community assemblages. To test this, we sampled three vertical transects along a steep rock wall at three depths (0 m, 4 m, 15 m), covering two distinct communities that were separated by near‐permanent water column stratification in the form of a strong halocline at ~3 m. Using three metabarcoding assays, our eDNA metabarcoding survey detected 54 taxa, across 46 families and 7 phyla, including 19 fish, 15 crustacean, and 8 echinoderm species. Ordination and cluster analyses show distinct eDNA signals across the halocline for all three replicate transects, suggesting that vertical dispersal of eDNA between communities was limited. Furthermore, eDNA signals of individual taxa were only retrieved within their observed vertical distribution, providing biological validation for the obtained results. Our results demonstrate, for the first time, the need to take into consideration oceanographic (e.g. water column stratification) and biological processes (e.g. vertical community structuring) when designing sampling strategies for marine eDNA metabarcoding surveys.

Abstract Each year, massive numbers of insects fly across the continents at heights of hundreds of meters, carried by the wind, bringing both environmental benefits and serious economic and social costs. To investigate the insects' flight behavior and their response to winds, entomological radar has proved to be a particularly valuable tool; however, its observations of insect orientation are ambiguous with regard to the head/tail direction, and this greatly hinders interpretation of the migrants' flight behavior. We have developed two related methods of using wind data to resolve the head/tail ambiguity, and we have compared their outputs with those from simply assigning the heading direction to be that which is closer to the track direction. We applied all three methods to observations of Australian plague locust migrations made with an insect monitoring radar. For the study dataset, some of the headings selected by the simpler method are shown to be clearly incorrect. The two new methods generally agree and reveal a significantly different, and presumably more accurate, relationship of heading direction to track direction. However, use of these methods leads to quite a large proportion of the sample being lost because the wind values, which derive from a regional‐scale numerical model, are shown to be incompatible with the radar observations. This exploratory study has moreover demonstrated that locusts are frequently oriented at a large angle to their track and that quite often their movement is at least slightly tailfirst. Both new methods appear to be a significant improvement on the simpler method. As well as providing an accurate representation of migratory flight behavior, they allow occasions when the model wind values are unreliable to be eliminated from the data sample.

Background This prospective study investigated the association between tooth loss and upper gastrointestinal (UGI) cancer mortality in the Linxian Dysplasia Nutrition Intervention Trial Cohort. Methods Subjects were categorized into three groups according to age at baseline. No missing teeth and less or greater than median tooth loss in each group was defined as none, moderate, and severe, respectively. Hazard ratios (HRs) and 95% confidence intervals (95% CIs) were estimated using the Cox proportional hazard model. Results Through 30 September 2015, 541 esophageal squamous cell carcinoma (ESCC), 284 gastric cardia carcinoma (GCC), and 77 gastric non‐cardia carcinoma (GNCC) deaths occurred. In the six‐year follow‐up, severe tooth loss was associated with an increased risk of GCC mortality (HR 1.55, 95% CI 1.06–2.18). In the 15‐year follow‐up, moderate tooth loss increased the ESCC mortality risk by 58% (HR 1.58, 95% CI 1.06–2.35), while severe loss increased the GCC mortality risk by 30% (HR 1.30, 95% CI 1.03–1.64). In the 30‐year follow‐up, moderate tooth loss increased the risk of ESCC mortality (HR 1.34, 95% CI 1.01–1.76). In subjects aged < 55 at baseline and men, moderate tooth loss had 53% and 52% higher risks of ESCC mortality (HR

Abstract DNA extraction from environmental samples (environmental DNA; eDNA) for metabarcoding‐based biodiversity studies is gaining popularity as a noninvasive, time‐efficient, and cost‐effective monitoring tool. The potential benefits are promising for marine conservation, as the marine biome is frequently under‐surveyed due to its inaccessibility and the consequent high costs involved. With increasing numbers of eDNA‐related publications have come a wide array of capture and extraction methods. Without visual species confirmation, inconsistent use of laboratory protocols hinders comparability between studies because the efficiency of target DNA isolation may vary. We determined an optimal protocol (capture and extraction) for marine eDNA research based on total DNA yield measurements by comparing commonly employed methods of seawater filtering and DNA isolation. We compared metabarcoding results of both targeted (small taxonomic group with species‐level assignment) and universal (broad taxonomic group with genus/family‐level assignment) approaches obtained from replicates treated with the optimal and a low‐performance capture and extraction protocol to determine the impact of protocol choice and DNA yield on biodiversity detection. Filtration through cellulose‐nitrate membranes and extraction with Qiagen's DNeasy Blood & Tissue Kit outperformed other combinations of capture and extraction methods, showing a ninefold improvement in DNA yield over the poorest performing methods. Use of optimized protocols resulted in a significant increase in OTU and species richness for targeted metabarcoding assays. However, changing protocols made little difference to the OTU and taxon richness obtained using universal metabarcoding assays. Our results demonstrate an increased risk of false‐negative species detection for targeted eDNA approaches when protocols with poor DNA isolation efficacy are employed. Appropriate optimization is therefore essential for eDNA monitoring to remain a powerful, efficient, and relatively cheap method for biodiversity assessments. For seawater, we advocate filtration through cellulose‐nitrate membranes and extraction with Qiagen's DNeasy Blood & Tissue Kit or phenol‐chloroform‐isoamyl for successful implementation of eDNA multi‐marker metabarcoding surveys.

Abstract Monarch butterflies are a species of conservation priority due to declining overwintering populations in both eastern and western North America. Declines in western overwintering monarchs—more than 99.9% since monitoring began—are especially acute. However, the degree to which western monarchs are a distinct biological entity is uncertain. In this review, we focus on phenotypic and genetic differentiation between eastern and western monarchs, with the goal of informing researchers and policy‐makers who are interested in monarch conservation. Eastern and western monarchs occupy distinct environments and show some evidence for phenotypic differentiation, particularly for migration‐associated traits, though population genetic and genomic studies suggest that they are indistinguishable from one another. We suggest future studies that could improve our understanding of differences between eastern and western monarchs. We also discuss the concept of adaptive capacity in eastern and western monarchs as well as non‐migratory populations outside of the monarch's primary North American range. Finally, we discuss the prospect of completely losing migratory monarchs from western North America and what this entails for monarch conservation.

Abstract Urban landscapes combining trees and crops—urban agroforestry (UAF) systems—may offer greater ecological and cultural benefits than annual cropping systems. Interest in UAF is growing, as evidenced by an increasing number of built projects and articles in the popular press and the academic literature on the subject. However, the practice of UAF appears to far outpace research on its scientific underpinnings or its design. Developing sustainable, resilient UAF sites can be challenging because of biophysical and sociocultural conditions unique to the city; however, cities offer opportunities not found in rural environments including the potential to close open nutrient loops between consumers and sites of food production. We argue that these biophysical and sociocultural challenges and opportunities can be best addressed through an evidence‐based approach to the design of UAF systems and a complex ecological aesthetic design language integrating theory, principles, and practices from urban agroecology and allied fields, environmental psychology, and landscape architecture. The resulting multifunctional UAF systems would be socially sustainable and equitable and promote the circular metabolism of the city. Drawing on a purposive review of literature from these disciplines, we propose a preliminary framework consisting of 14 guidelines and complementary principles and strategies for the design of multifunctional, culturally preferred UAF and offer recommendations for future research.

Abstract This Technical Report describes the activities developed in the scope of the EU‐FORA Fellowship, within the work programme of risk assessment (RA) of exotic disease incursion and spread, developed at Wageningen Bioveterinary Research (WBVR). The programme focused on the work carried out in the Generic risk assessment for introduction of animal diseases (G‐RAID) project, which brings together a number of different generic RA tools from multiple European partners. The aim of the fellowship was to gain understanding of veterinary import risk assessment by using different RA tools and to learn how different algorithms can be used to calculate disease incursion risks. G‐RAID's tools cover a wide range of RA methodologies; from purely qualitative, to semi‐quantitative and fully stochastic quantitative methods, which allowed the fellow to understand a variety of algorithms used to produce the final risk estimate. The fellowship programme provided the fellow with the chance to learn in detail about how generic RAs are performed across Europe, understanding how to deal with the uncertainty and variability involved in RAs and the potential problems of data availability and reliability. The fellow made an inventory of publicly available databases on disease occurrence and international trade that could be used for import RA and assessed their quality and usefulness for the different generic RA tools. The programme also provided the fellow the opportunity to perform several import risk assessments using the RA tools of G‐RAID. She completed a RA on African swine fever using the MINTRISK model developed by WBVR. Furthermore, she assessed the risk of foot and mouth disease introduction using the Rapid Risk Assessment Tool (RRAT) model developed by WBVR and the COMPARE model developed by the Animal and Plant Health Agency (APHA). To this end, the fellow completed a short‐term visit to APHA, enabling her to have additional training in quantitative RA and to expand her professional network in this area.