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Introduction: Relatively little is known about the molecular pathways influenced by cannabis use in humans. We used a multi-omics approach to examine protein, metabolomic, and lipid markers in plasma differentiating between cannabis users and nonusers to understand markers associated with cannabis use. Methods: Eight discordant twin pairs and four concordant twin pairs for cannabis use completed a blood draw, urine and plasma toxicology testing, and provided information about their past 30-day cannabis use and other substance use patterns. The 24 twins were all non-Hispanic whites. Sixty-six percent were female. Median age was 30 years. Fifteen participants reported that they had used cannabis in the last 30 days, including eight participants that used every day or almost every day (29–30 of 30 days). Of these 15 participants, plasma 11-nor-9-carboxy-Δ(9)-tetrahydrocannabinol (THC-COOH) and total tetrahydrocannabinol (THC) concentrations were detectable in 12 participants. Among the eight “heavy users” the amount of total THC (sum of THC and its metabolites) and plasma THC-COOH concentrations varied widely, with ranges of 13.1–1713 ng/mL and 2.7–284 ng/mL, respectively. A validated liquid chromatography–tandem mass spectrometry (LC-MS/MS) assay measured plasma THC-COOH, THC, and other cannabinoids and metabolites. Plasma THC-COOH was used as the primary measure. Expression levels of 1305 proteins were measured using SOMAScan assay, and 34 lipid mediators and 314 metabolites were measured with LC-MS/MS. Analyses examined associations between markers and THC-COOH levels with and without taking genetic relatedness into account. Results: Thirteen proteins, three metabolites, and two lipids were identified as associated with THC-COOH levels. Myc proto-oncogene was identified as associated with THC-COOH levels in both molecular insight and potential marker analyses. Five pathways (interleukin-6 production, T lymphocyte regulation, apoptosis, kinase signaling pathways, and nuclear factor kappa-light-chain-enhancer of activated B cells) were linked with molecules identified in these analyses. Conclusions: THC-COOH levels are associated with immune system-related pathways. This study presents a feasible approach to identify additional molecular markers associated with THC-COOH levels.

OBJECTIVES: Cannabis legalization may have unintended consequences, such as effects on prevalence of adolescent use of other substances. We hypothesize that in states that have legalized recreational cannabis (RCL states), electronic vapor product use (“vaping”), cannabis, and cigarette use among adolescents is more prevalent than in states that have only legalized medical cannabis(MCL states) and states that have neither legalized recreational nor medical cannabis (NL states). METHODS: Data are from the Center for Disease Control’s 2017 Youth Risk Behavior Surveillance Survey (n=107,665), a nationally representative survey of US high school students. We categorized student responses by cannabis legalization status of the state in which they lived at time of survey, then tested associations between cannabis legalization status and adolescent vaping, cannabis, and cigarette use, using logistic regressions. RESULTS: Students in RCL states were significantly more likely to report current vaping behaviors compared to NL students [odds ratios (OR’s) 2.07–2.21]. Students in cannabis-legal states were significantly more likely to report ever or currently using cannabis compared to NL students (OR’s 1.27–1.40). MCL students were significantly less likely to report current cigarette smoking (OR=0.86) compared to NL students. CONCLUSIONS: Adolescents in cannabis-legal states were more likely to report vaping and cannabis use compared to adolescents in NL states. Public health officials, policymakers, and clinicians should consider the associations between cannabis legalization and adolescent vaping, cigarette, and cannabis use patterns.

The incidence of tickborne diseases in the United States is increasing; reported cases more than doubled from >22,000 in 2004 to >48,000 in 2016 (1). Ticks are responsible for approximately 95% of all locally acquired vectorborne diseases reported by states and the District of Columbia, with Lyme disease accounting for >80% of those cases (2). After a tick bite, persons might seek care at an emergency department (ED) for tick removal and to receive postexposure prophylaxis, which has been shown to effectively prevent Lyme disease when taken within 72 hours of a high-risk bite (3). Using data from CDC's National Syndromic Surveillance Program (NSSP), investigators examined ED tick bite visits during January 2017-December 2019 by sex, age group, U.S. region, and seasonality. During this 36-month period, 149,364 ED tick bite visits were identified. Mean cumulative incidence was 49 ED tick bite visits per 100,000 ED visits overall; incidence was highest in the Northeast (110 per 100,000 ED visits). The seasonal distribution of ED tick bite visits was bimodal: the larger peak occurred during the spring and early summer, and the smaller peak occurred in the fall. This pattern aligns with the seasonality of a known and abundant human-biter, the blacklegged tick, Ixodes scapularis (4). Compared with other age groups, pediatric patients aged 0-9 years accounted for the highest number and incidence of ED tick bite visits; incidence was higher among male patients than among females. Tick bites are not monitored by current surveillance systems because a tick bite is an event that in and of itself is not a reportable condition to health departments. Syndromic surveillance of ED tick bite visits can provide timely information that might predict temporal and geographic risk for exposure to tickborne diseases and guide actionable public health messaging such as avoiding tick habitats, wearing repellent consistently when outdoors, and performing regular tick checks during times of increased tick bite risk.

Surveys indicate US residents spent more time outdoors in 2020 than in 2019, but fewer tick bite-related emergency department visits and Lyme disease laboratory tests were reported. Despite ongoing exposure, Lyme disease case reporting for 2020 might be artificially reduced due to coronavirus disease-associated changes in healthcare-seeking behavior.

Objectives Although tick-borne diseases account for a large number of health care visits in the United States, clinical practices for tick bite and Lyme disease treatment and prevention are not well understood. The objective of this study was to better understand factors associated with clinical practices related to tick bites and Lyme disease. Methods In 2013-2015, questions about tick-bite evaluation, Lyme disease diagnosis and treatment, appropriate use of Lyme disease testing, and tick-bite prevention were included in Porter Novelli’s DocStyles survey, a nationally representative annual web-based survey of health care providers. We performed analyses of responses by provider license type and state-level incidence (high or low) of Lyme disease in 2019. Results A total of 4517 providers were surveyed across the 3 study years. Overall, 80.9% of providers reported that they had evaluated at least 1 patient for a tick bite, 47.6% had diagnosed at least 1 patient with Lyme disease, and 61.9% had treated at least 1 patient for Lyme disease in the previous year. Providers from states with a high incidence of Lyme disease saw more patients for tick bites and Lyme disease than providers from states with a low incidence of Lyme disease. Few providers correctly chose Lyme disease testing as clinically useful in the hypothetical case of a patient from a state with a high incidence of Lyme disease with an arthritic knee (36.0%) or with new-onset atrioventricular block (39.5%), and respondents across all provider types incorrectly chose testing when not clinically indicated. Most providers (69.7%) reported routinely recommending tick-bite prevention methods to patients. Conclusions Many providers evaluate patients for tick bites and treat patients for Lyme disease, but knowledge about appropriate testing is low. Providers may benefit from tailored education about appropriate Lyme disease diagnosis, testing, and effective tick-bite prevention.

By using commercial insurance claims data, we estimated that Lyme disease was diagnosed and treated in ≈476,000 patients in the United States annually during 2010-2018. Our results underscore the need for accurate diagnosis and improved prevention.

Voltage-gated sodium channels (Navs) are promising targets for analgesic and antiepileptic therapies. Although specificity between Nav subtypes may be desirable to target specific neural types, such as nociceptors in pain, many broadly acting Nav inhibitors are clinically beneficial in neuropathic pain and epilepsy. Here, we present the first systematic characterization of vixotrigine, a Nav blocker. Using recombinant systems, we find that vixotrigine potency is enhanced in a voltage- and use-dependent manner, consistent with a state-dependent block of Navs. Furthermore, we find that vixotrigine potently inhibits sodium currents produced by both peripheral and central nervous system Nav subtypes, with use-dependent IC50 values between 1.76 and 5.12 μM. Compared with carbamazepine, vixotrigine shows higher potency and more profound state-dependent inhibition but a similar broad spectrum of action distinct from Nav1.7- and Nav1.8-specific blockers. We find that vixotrigine rapidly inhibits Navs and prolongs recovery from the fast-inactivated state. In native rodent dorsal root ganglion sodium channels, we find that vixotrigine shifts steady-state inactivation curves. Based on these results, we conclude that vixotrigine is a broad-spectrum, state-dependent Nav blocker. SIGNIFICANCE STATEMENT: Vixotrigine blocks both peripheral and central voltage-gated sodium channel subtypes. Neurophysiological approaches in recombinant systems and sensory neurons suggest this block is state-dependent.

The hyperdiverse shrew genus Crocidura is one of few small mammal genera distributed across Sundaland and all of its boundaries. This represents a rare opportunity to study the geological history of this region through the evolutionary history of these shrews. We generate a phylogeny of all recognized species of Sundaland Crocidura and show that most speciation events took place during the Pleistocene, prior to the inundation of the Sunda Shelf around 400 000 years ago. We find east–west differentiation within two separate lineages on Borneo, and that the current taxonomy of its two endemic species does not reflect evolutionary history, but ecophenotypic variation of plastic traits related to elevation. Sulawesi shrews are monophyletic, with a single notable exception: the black-footed shrew (C. nigripes). We show that the black-footed shrew diverged from its relatives on Borneo recently, suggesting a human-assisted breach of Wallace’s line. Overall, the number of Crocidura species, especially on Borneo, probably remains an underestimate.

Since 1970, >50% of patients with plague in the United States had interactions with animals that might have led to infection. Among patients with pneumonic plague, nearly all had animal exposure. Improved understanding of the varied ways in which animal contact might increase risk for infection could enhance prevention messages.

Manipulating hydrologic conditions at the land-water interface is critical for managing wetland functions. Hydrologic manipulation to increase retention time of water is used to promote wetland conditions, enhance nitrogen (N) processing for reduced N export, and attract migratory bird populations. Human managed wetlands such as waterfowl impoundments are intended to attract waterfowl for tourism. The limited literature has shown that waterfowl impoundments export N during seasonally prescribed drawdowns; however, it is unknown how impoundment-specific characteristics and different types of impoundments influence N cycling transformations. We compared seasonal N cycling between and within moist-soil managed (MSM) and agricultural (Ag) waterfowl impoundment soils. Potential nitrification, denitrification, and N mineralization rates and soil physicochemical properties were analyzed. Potential nitrification and denitrification rates were higher in the Ag compared to MSM impoundment even when the MSM site is actively managed to promote wetland conditions year-round. Despite the higher soil organic carbon and soil moisture content at MSM compared to Ag site, the high extractable soil ammonium, low nitrate, and low nitrification rates at MSM are evidence of substrate limitation for denitrification but not nitrification. These results indicate that decoupling of nitrification and denitrification could explain the reduced N removal capacity in these managed wetlands.

We present the results of a study of the connectivity between Pacific cod spawning and nursery areas, and settlement of Pacific cod in the Gulf of Alaska. This work was conducted to address the hypothesis that spatial and temporal patterns of recruitment are related to variability in connectivity between spawning and nursery areas. To examine this hypothesis, we developed a Lagrangian, biophysical, individual-based model of Pacific cod early life history and dispersal using the Dispersal Model for Early Life Stages (DisMELS) framework. This model is driven by currents and scalars such as temperature from a version of the Regional Oceanographic Model System (ROMS) developed for the Gulf of Alaska. Results of our study show connectivity patterns predicted by the model that agree with our understanding (based on genetic analyses) that there is a high degree of localized retention in Pacific cod. The results indicate that the Shumagin Islands and Prince William Sound regions may serve as important collectors of Pacific cod recruits from upstream spawning areas. We also find correlations between individual-based model outputs and several large-scale climate indicators that appear to show settlement in several important nursery areas, and recruitment overall, are positively affected by slower gyre circulation in the Gulf of Alaska. We hypothesize that this is due to enhancement of retention, settlement in the Shumagin Island region, and reduction of transport of young cod out of the Gulf of Alaska to the southwest.

Little is known about the mechanism of transport that enables age-0 sablefish (Anoplopoma fimbria) to reach suitable nursery sites from spawning locations far offshore, or the strength of the connection between individual spawning sites and nursery areas, or how variability in the strength of these connections may impact recruitment success. Using a model for the early life stages of sablefish, we explored the variability in connectivity between spawning and recruitment sites that can arise solely from interannual variability in environmental forcing and its impact on transport. Our major findings are that 1) the model indicates young sablefish settling in nursery areas in the Gulf of Alaska were most likely spawned in the eastern Gulf; 2) sablefish spawned in the western Gulf of Alaska are unlikely to settle anywhere in the Gulf, and are more likely to be advected farther west, perhaps to settle in the Aleutian islands or Bering Sea (to contribute to the Alaska population, they would have to undergo an active return migration as they mature); 3) total connectivity between all spawning sites and nursery areas showed stronger correlation with recruitment estimates than the strength of connections to or from specific regions; and 4) transport to St. John Baptist Bay, a known sablefish nursery area, was not the most probable end point for sablefish spawned throughout our Gulf of Alaska model domain. This suggests that young individuals arrive at this persistent nursery area due to directional swimming behavior, highly localized spawning, or small-scale currents not captured in the hydrographic model. The fact that no single correlate in our analysis had a very strong relationship to sablefish recruitment indicates that recruitment variability arises from complex interactions between the environment and the individual, and a possible disconnect in spatial scales between the Gulf of Alaska sablefish IBM and the broader sablefish stock assessment, which includes both the GOA and the Eastern Bering Sea, as well as possible contributions from Canadian stocks to the south. Our analyses determined that although the timing and extent of this transport shows significant interannual variability, both the location of likely sablefish spawning (source) areas and the comparative strength of connectivity between spawning and nursery sites appear to be relatively consistent year-to-year.

Little is known regarding the importance of early-life transport and dispersion mechanisms in determining recruitment variability for Pacific ocean perch (POP) in the Gulf of Alaska (GOA). These mechanisms influence the degree of, and variability in, connectivity between offshore natal areas (parturition sites) and inshore demersal nursery habitats for young-of-the year juveniles, and may thus play an important role in the “gauntlet” that individuals must survive from parturition to recruitment. As a first attempt to assess interannual variability in connectivity between natal and nursery areas for POP in the GOA in a synthetic manner, we developed a coupled biophysical individual-based model (IBM) for POP early life history and dispersal with simple representations of active vertical movement, passive horizontal movement, growth, and settlement in appropriate nursery habitat to integrate known early-life traits with variability in environmental forcing. We used an oceanographic model for the GOA based on the Regional Ocean Modeling system (ROMS) to provide the underlying daily physical environment to force the IBM for 1996–2011 and simulated hundreds of thousands of individual POP from parturition along the shelf break to settlement in inshore demersal nursery habitats as young-of-the-year. We used the IBM results to assess spatial patterns of annual “maximum potential” connectivity between presumed natal and nursery areas at alongshore scales of ~ 150 km, as well as the interannual variability in these patterns. Results showed that, even in the absence of mortality, most (> 70%) individuals were unsuccessful in dispersing from presumed natal areas along the continental shelf break to inshore nursery areas. For those that were successful, connectivity was directed in a counterclockwise fashion (southeast to northwest) around the GOA following prevailing current patterns. Typical dispersion distances were on the order of 100's of km alongshore, much larger than those inferred from genetic sampling. Natal areas from which the highest fractions of successful individuals originated were in the southeast GOA, while the nursery areas most frequently reached by those successful individuals were in the central GOA. POP from natal areas in the western GOA were consistently exported from the system and likely contribute little to the GOA population, although they may contribute to populations in the Aleutian Islands and eastern Bering Sea. We also found that annual indices derived from the connectivity matrices were not very strongly related to any of a suite of basin- and regional-scale environmental indices, reflecting the overall complexity and scale of the pathways POP in the GOA may undertake during their early life stages and suggesting that multiple drivers operating at different spatial and temporal scales influence connectivity patterns. Finally, while our results indicate that interannual variability in physical transport may have a substantial impact on connectivity, we found little support for the hypothesis that this alone drives variability in POP recruitment.

Tick-borne relapsing fever (TBRF) is a potentially serious spirochetal infection caused by certain species of Borrelia and acquired through the bite of Ornithodoros ticks. In 2017, Austin Public Health, Austin, TX, identified five cases of febrile illness among employees who worked in caves. A cross-sectional serosurvey and interview were conducted for 44 employees at eight organizations that conduct cave-related work. Antibodies against TBRF-causing Borrelia were detected in the serum of five participants, four of whom reported recent illness. Seropositive employees entered significantly more caves (Median 25 [SD: 15] versus Median 4 [SD: 16], p = 0.04) than seronegative employees. Six caves were entered more frequently by seropositive employees posing a potentially high risk. Several of these caves were in public use areas and were opened for tours. Education of area healthcare providers about TBRF and prevention recommendations for cavers and the public are advised.

In July 2017, fever and sepsis developed in 3 recipients of solid organs (1 heart and 2 kidneys) from a common donor in the United States; 1 of the kidney recipients died. Tularemia was suspected only after blood cultures from the surviving kidney recipient grew Francisella species. The organ donor, a middle-aged man from the southwestern United States, had been hospitalized for acute alcohol withdrawal syndrome, pneumonia, and multiorgan failure. F. tularensis subsp. tularensis (clade A2) was cultured from archived spleen tissue from the donor and blood from both kidney recipients. Whole-genome multilocus sequence typing indicated that the isolated strains were indistinguishable. The heart recipient remained seronegative with negative blood cultures but had been receiving antimicrobial drugs for a medical device infection before transplant. Two lagomorph carcasses collected near the donor's residence were positive by PCR for F. tularensis subsp. tularensis (clade A2). This investigation documents F. tularensis transmission by solid organ transplantation.

Translation includes initiation, elongation, and termination, followed by ribosome recycling. We characterize a new sequence element in 5’ untranslated regions that consists of an adjacent start and stop codon and thereby excludes elongation. In these start-stop elements, an initiating ribosome is simultaneously positioned for termination without having translocated. At the example of activating transcription factor 4 (ATF4), we demonstrate that start-stops modify downstream re-initiation, thereby repressing translation of upstream open reading frames and enhancing ATF4’s inducibility under stress. Start-stop elements are abundant in both mammals and yeast and affect key regulators such as DROSHA and the oncogenic transcription factor NFIA. They provide a unique regulatory layer that impedes ribosome scanning without the energy-expensive peptide production that accompanies upstream open reading frames.One-Sentence SummaryRegulatory start-stop elements in the 5’UTR are hitherto unappreciated contributors to 5’ UTR code and alter the translatome.

During a recent outbreak of Bartonella quintana disease in Denver, 15% of 241 persons experiencing homelessness who presented for severe acute respiratory syndrome coronavirus 2 testing were seroreactive for Bartonella. Improved recognition of B quintana disease and prevention of louse infestation are critical for this vulnerable population.

Tick bite prevention practices, knowledge of Lyme disease (LD) symptoms and transmission, and patterns of LD diagnoses among Hispanic persons have been reported but not comprehensively evaluated. In 2014, CDC examined questions from a prospective nationwide survey of U.S. Hispanic adults conducted via the Offerwise QueOpinas panel regarding ticks and LD. From October to November, a total of 2,649 surveys were released and 1,006 completed surveys returned. Overall, 44% of respondents reported routinely practising at least one form of personal protection against tick bites, and wearing repellent was the most commonly reported method (29%). Approximately 6% of respondents reported a tick bite for either themselves or someone in their household during the previous 12 months. An individual or household diagnosis of LD in the previous year was reported by 2% of respondents, with the highest proportion of diagnoses reported by respondents from high LD incidence states. The annual incidence of healthcare provider-diagnosed LD in the survey population was higher than national surveillance estimates for reported LD among U.S. Hispanic persons during 2000-2013. As annual incidence of LD continues to increase, it is important to ensure equitable access to information about LD, including disease transmission, manifestations, and prevention recommendations. Results from this survey can help inform public health outreach focused on effective tick bite prevention methods and early recognition of LD.

Tickborne diseases are an increasing public health problem in the northeastern USA. Bait boxes that apply acaricide to rodents have been shown in small field studies to significantly reduce abundance of Ixodes scapularis ticks as well as their pathogen infection rates in treated areas. The effectiveness of this intervention for preventing human tickborne diseases (TBDs) has not been demonstrated. During 2012-2016, TickNET collaborators conducted a randomized, blinded, placebo-controlled trial among 622 Connecticut households. Each household received active (containing fipronil wick) or placebo (empty) bait boxes in their yards over two consecutive years. Information on tick encounters and TBDs among household members was collected through biannual surveys. Nymphal ticks were collected from a subset of 100 properties during spring at baseline, during treatment, and in the year post-intervention. Demographic and property characteristics did not differ between treatment groups. There were no significant differences post-intervention between treatment groups with respect to tick density or pathogen infection rates, nor for tick encounters or TBDs among household members. We found no evidence that rodent-targeted bait boxes disrupt pathogen transmission cycles or significantly reduce household risk of tick exposure or TBDs. The effectiveness of this intervention may depend on scale of use or local enzootic cycles.

Identifying molecular mediators of neural circuit development and/or function that contribute to circuit dysfunction when aberrantly reengaged in neurological disorders is of high importance. The role of the TWEAK/Fn14 pathway, which was recently reported to be a microglial/neuronal axis mediating synaptic refinement in experience-dependent visual development, has not been explored in synaptic function within the mature central nervous system. By combining electrophysiological and phosphoproteomic approaches, we show that TWEAK acutely dampens basal synaptic transmission and plasticity through neuronal Fn14 and impacts the phosphorylation state of pre- and postsynaptic proteins in adult mouse hippocampal slices. Importantly, this is relevant in two models featuring synaptic deficits. Blocking TWEAK/Fn14 signaling augments synaptic function in hippocampal slices from amyloid-beta–overexpressing mice. After stroke, genetic or pharmacological inhibition of TWEAK/Fn14 signaling augments basal synaptic transmission and normalizes plasticity. Our data support a glial/neuronal axis that critically modifies synaptic physiology and pathophysiology in different contexts in the mature brain and may be a therapeutic target for improving neurophysiological outcomes.

Entomological measures have long served as proxies for human risk of Lyme disease (LD) and other tickborne diseases (TBDs) in endemic areas of the United States, despite conflicting results regarding the correlation between these measures and human disease outcomes. Using data from a previous TBD intervention study in Connecticut, Maryland and New York, we evaluated whether human-tick encounters can serve as an accurate proxy for risk of TBDs in areas where LD and other Ixodes scapularis-transmitted infections are common. Among 2,590 households consisting of 4,210 individuals, experiencing a tick encounter was associated with an increased risk of both self-reported (RR = 3.17, 95% CI: 2.05, 4.91) and verified TBD (RR = 2.60, 95% CI: 1.39, 4.84) at the household level. Household characteristics associated with experiencing any tick encounter were residence in Connecticut (aOR = 1.86, 95% CI: 1.38, 2.51) or New York (aOR = 1.66, 95% CI: 1.25, 2.22), head of household having a graduate level education (aOR = 1.46, 95% CI: 1.04, 2.08), owning a pet (aOR = 1.80, 95% CI: 1.46, 2.23) and a property size of 2 acres or larger (aOR = 2.30, 95% CI: 1.42, 3.70). Results for individual characteristics were similar to those for households. Future prevention studies in LD endemic areas should consider using human-tick encounters as a robust proxy for TBD risk.

West Nile Virus (WNV) is a positive polarity, single-stranded RNA virus that causes West Nile fever, for which no cure has been found to date. WNV, like other RNA viruses, needs to compact all the information to complete the viral cycle into a very small genome. Beyond the information that is stored in the primary structure, the genome of RNA viruses bear functional structural domains that perform multiple essential functions for the viral cycle. In WNV, several of these functional domains are found in the 3'UTR region. Based on the importance of these functional domains, in this work, RNA aptamers have been studied as a possible therapeutic agent. Aptamers are oligonucleotides with the ability to efficiently bind to a molecule, not taking into account only the sequence of the target but also its structural motifs. In this work, various aptamers directed against the 3'UTR region of WNV, which could potentially inhibit processes of the WNV viral cycle, have been analysed and selected by in silico analysis. We have also studied certain characteristics of the SL-I structural element of the WNV 3¿UTR, which shows a high chance of interacting with host molecules. This work will lead further studies towards the generation of antiviral aptamers against WNV and a deeper understanding of WNV interaction with the host cell.

In plants, the vegetative to reproductive phase transition (termed bolting in Arabidopsis) generally precedes age‐dependent leaf senescence (LS). Many studies describe a temporal link between bolting time and LS, as plants that bolt early, senesce early, and plants that bolt late, senesce late. The molecular mechanisms underlying this relationship are unknown and are potentially agriculturally important, as they may allow for the development of crops that can overcome early LS caused by stress‐related early‐phase transition. We hypothesized that leaf gene expression changes occurring in synchrony with bolting were regulating LS. ARABIDOPSIS TRITHORAX (ATX) enzymes are general methyltransferases that regulate the adult vegetative to reproductive phase transition. We generated an atx1, atx3, and atx4 (atx1,3,4) triple T‐DNA insertion mutant that displays both early bolting and early LS. This mutant was used in an RNA‐seq time‐series experiment to identify gene expression changes in rosette leaves that are likely associated with bolting. By comparing the early bolting mutant to vegetative WT plants of the same age, we were able to generate a list of differentially expressed genes (DEGs) that change expression with bolting as the plants age. We trimmed the list by intersection with publicly available WT datasets, which removed genes from our DEG list that were atx1,3,4 specific. The resulting 398 bolting‐associated genes (BAGs) are differentially expressed in a mature rosette leaf at bolting. The BAG list contains many well‐characterized LS regulators (ORE1, WRKY45, NAP, WRKY28), and GO analysis revealed enrichment for LS and LS‐related processes. These bolting‐associated LS regulators may contribute to the temporal coupling of bolting time to LS.

A surprising amount of hidden phylogenetic diversity exists in the small to medium size, drab colored squirrels of the genus Sundasciurus. This genus is endemic to Sundaland and the Philippines, where it is widespread. An earlier revision of this genus found that the high elevation ‘populations’ of the widespread, lowland slender squirrel (S. tenuis) were different species. Previous phylogenies based on mitochondrial cytochrome b sequences also suggested that the widespread, lowland Low’s squirrel (S. lowii) and the narrow endemic Fraternal squirrel (S. fraterculus) are not reciprocally monophyletic. Additionally, deep divergences have been identified between lineages within Low’s squirrel that date to the early Pliocene. Here we focus on evaluating the relationships and differences within and between populations of these two nominal species using whole mitochondrial genome sequences, nuclear intron sequences, and morphology. We reassess the taxonomy of this group, revalidate the species status of Robinson’s squirrel (Sundasciurus robinsoni Bonhote, 1903) support the species level recognition of the Natuna squirrel (Sundasciurus natunensis Thomas, 1895) and identify three other lineages that require further study. We estimate times of divergence and integrate geologic history to find that most of the divergences are pre-Pleistocene, and thus predate the Pleistocene flooding of Sundaland. Biogeographic, and ecological factors may have played a more important role than climatic factors in generating these patterns. While divergence in allopatry seems to be the main process driving speciation in lowland Sundaland squirrels (Sundasciurus), ecomorphological and behavioral adaptations in this clade suggest an important role of niche divergence., AH was supported by an Ernst Mayr Travel grant and AMNH Collection Study Grant during museum data collection. This research received support from the SYNTHESYS Project financed by European Community Research Infrastructure Action under the FP7 “Capacities” Program. AH was also supported by a Spanish Ministry of Economy contract CGL2014-58793-P. The Spanish Ministry of Science and Innovation grant CGL2017-86068-P to JL supported this study. MH was supported by discretionary research funding from Humboldt State University.

Consistent and effective use of personal prevention methods for tickborne diseases, including Lyme disease (LD), is dependent on risk awareness. To improve our understanding of the general U.S. population's experiences with tick exposure and use of personal prevention methods, we used data from ConsumerStyles, a web-based, nationally representative questionnaire on health-related topics. Questions addressed tick bites and LD diagnosis in the last year, use of personal prevention methods to prevent tick bites, and willingness to receive a theoretical LD vaccine. Of 10,551 participants surveyed over three years, 12.3 % reported a tick bite for themselves or a household member in the last year, including 15.4 % of participants in high LD incidence (LDI) states, 16.3 % in states neighboring high LDI states, and 9.4 % in low LDI states. Participants in high LDI states and neighboring states were most likely to use personal prevention methods, though 46.6 % of participants in high LDI states and 53.9 % in neighboring states reported not using any method. Participants in low LDI states, adults ≥ 75 years of age, those with higher incomes, and those living in urban housing tended to be less likely to practice personal prevention methods. Likeliness to receive a theoretical LD vaccine was high in high LDI (64.5 %), neighboring (52.5 %), and low LDI (49.7 %) states. Targeted educational efforts are needed to ensure those in high LDI and neighboring states, particularly older adults, are aware of their risk of LD and recommended personal prevention methods.

The original version of this article unfortunately contained an error in the areas of the impoundments reported in Section 2.1 of the published article.

Background Social impairments are important features of a substance use disorder diagnosis; and recent models suggest early impairments in socio-cognitive and -affective processes may predict future use. However, no systematic reviews are available on this topic. Methods We conducted a systematic review and meta-analyses exploring the association between social-cognitive and -affective processes (empathy, callous-unemotional (CU) traits, theory of mind, and social cognition) and substance use frequency (alcohol, cannabis, general drug use). We examined moderating effects of study design, gender, age, and weather conduct problems were controlled for. We also review brain studies related to social cognition and substance use disorder (SUD) risk. Results Systematic review suggested a negative association for positively valenced constructs with substance use but mixed results on the negatively valenced construct CU traits. Meta-analyses revealed moderate positive association between CU traits with alcohol and general drug use but no significance with cannabis use. Moderate effect sizes were found for CU traits in youth predicting severity of substance use by late adolescence and significantly accounted for variance independently of conduct problems. Significant moderators included gender proportions, sample type, and age. Neuroimaging meta-analysis indicated 10 coordinates that were different in youth at a high risk/with SUD compared to controls. Three of these coordinates associate with theory of mind and social cognition. Conclusion Socio-cognitive and -affective constructs demonstrate an association with current and future substance use, and neural differences are present when performing social cognitive tasks in regions with strongest associations with theory of mind and social cognition.

Kelp forests dominate wave-exposed rocky reefs along mid-to-high latitude coastlines. The distribution and structure of kelp forests is determined by a range of physical and biological processes operating across varying spatial and temporal scales. Many kelp forest systems are strongly influenced by herbivory, and overgrazing by urchins, in particular, is a recognised driver of kelp forest distribution and structure. The ecological significance of herbivory by kelp-associated limpets, however, has received far less attention. We quantified seasonal population dynamics of Laminaria digitata and the blue-rayed limpet Patella pellucida on two rocky shores in North Devon, UK. For the kelp, we quantified density, percent cover, morphology, standing biomass, elongation and erosion rates, and C:N and phenol content. For the limpet, we measured abundance, biomass, and morphology, and for the limpet-kelp interaction we recorded observable grazing damage on the blade. Both populations exhibited typical seasonality with kelp growth rates peaking in spring, maximum standing biomass observed in summer and increased erosion rates in autumn. Blue-rayed limpets, which were recorded within kelp holdfasts as well as on stipes and blades, recruited in spring and peaked in abundance in late summer, with length and biomass generally increasing through the year. The area of kelp blade damaged by limpet grazing was low (a maximum of ~4%) and relatively consistent throughout the year, and the number and size of grazing scars exhibited limited seasonality. Survey data from other L. digitata populations in the UK suggested that the limpet abundances recorded in North Devon were comparable with other sites in the region. Although the impact of P. pellucida grazing on kelp blade tissue appears to be minimal, further research into cavity grazing by limpets on the stipe and in the holdfast is needed to formerly assess the impact these cavities have on dislodgement and fracture rates, especially when increased occupancy and grazing coincides with periods of intense wave action. We conclude that while herbivory is an important processes acting upon many kelp populations globally, the grazing pressure exerted by P. pellucida on L. digitata is unlikely to strongly influence population structure on UK rocky shores.

Introduction Vectorborne diseases are major causes of death and illness worldwide. In the United States, the most common vectorborne pathogens are transmitted by ticks or mosquitoes, including those causing Lyme disease; Rocky Mountain spotted fever; and West Nile, dengue, and Zika virus diseases. This report examines trends in occurrence of nationally reportable vectorborne diseases during 2004-2016. Methods Data reported to the National Notifiable Diseases Surveillance System for 16 notifiable vectorborne diseases during 2004-2016 were analyzed; findings were tabulated by disease, vector type, location, and year. Results A total 642,602 cases were reported. The number of annual reports of tickborne bacterial and protozoan diseases more than doubled during this period, from >22,000 in 2004 to >48,000 in 2016. Lyme disease accounted for 82% of all tickborne disease reports during 2004-2016. The occurrence of mosquitoborne diseases was marked by virus epidemics. Transmission in Puerto Rico, the U.S. Virgin Islands, and American Samoa accounted for most reports of dengue, chikungunya, and Zika virus diseases; West Nile virus was endemic, and periodically epidemic, in the continental United States. Conclusions and implications for public health practice Vectorborne diseases are a large and growing public health problem in the United States, characterized by geographic specificity and frequent pathogen emergence and introduction. Differences in distribution and transmission dynamics of tickborne and mosquitoborne diseases are often rooted in biologic differences of the vectors. To effectively reduce transmission and respond to outbreaks will require major national improvement of surveillance, diagnostics, reporting, and vector control, as well as new tools, including vaccines.

Prevention of tick-borne diseases requires an understanding of when and where exposure to ticks is most likely. We used an epidemiologic approach to define these parameters for residents of a Lyme-endemic region. Two persons in each of 500 Connecticut households were asked to complete a log each night for one week during June, 2013. Participants recorded their whereabouts in 15min increments (indoors, outdoors in their yard, outdoors on others' private property, or outdoors in public spaces) and noted each day whether they found a tick on themselves. Demographic and household information was also collected. Logs were completed for 934 participants in 471 households yielding 51,895 time-place observations. Median participant age was 49 years (range 2-91 years); 52% were female. Ninety-one participants (9.8%) reported finding a tick during the week, with slightly higher rates among females and minors. Household factors positively associated with finding a tick included having indoor/outdoor pets (odds ratio (OR)=1.7; 95% confidence interval (CI): 1.1-2.9), the presence of a bird feeder in the yard (OR=1.9; CI:1.2-3.2), and presence of an outdoor dining area (OR=2.2; CI:1.1-4.3). Individual factors associated with finding a tick on a given day were bathing or showering (OR=3.7; CI:1.3-10.3) and hours spent in one's own yard (OR=1.2, CI:1.1-1.3). Nineteen participants found ticks on multiple days, more than expected assuming independence (p0.001). Participants who found ticks on multiple days did not spend more time outdoors but were significantly more likely to be male than those finding ticks on a single day (p0.03). Our findings suggest that most tick exposures in the study area occurred on private property controlled by the respective homeowner. Interventions that target private yards are a logical focus for prevention efforts.

Of the four human Argonaute (AGO) paralogs, only AGO2 has been shown to have slicer activity. The others (AGO1, AGO3 and AGO4) have been thought to assemble with microRNAs to form slicer-independent effector complexes that bind target mRNAs and silence gene expression through translational repression and deadenylation but not cleavage. Here, we report that recombinant AGO3 loaded with miR-20a cleaves complementary target RNAs, whereas AGO3 loaded with let-7a, miR-19b or miR-16 does not, indicating that AGO3 has slicer activity but that this activity depends on the guide RNA. Our cleavage assays using chimeric guides revealed the significance of seed sequence for AGO3 activity, which depends specifically on the sequence of the post-seed. Unlike AGO2, target cleavage by AGO3 requires both 5′- and 3′-flanking regions. Our 3.28 Å crystal structure shows that AGO3 forms a complete active site mirroring that of AGO2, but not a well-defined nucleic acid-binding channel. These results demonstrating that AGO3 also has slicer activity but with more intricate substrate requirements, explain the observation that AGO3 has retained the necessary catalytic residues throughout its evolution. In addition, our structure inspires the idea that the substrate-binding channel of AGO3 and consequently its cellular function, may be modulated by accessory proteins.

We examined whether pet ownership increased the risk for tick encounters and tick-borne disease among residents of three Lyme disease-endemic states as a nested cohort within a randomized controlled trial. Information about pet ownership, use of tick control for pets, property characteristics, tick encounters and human tickborne disease were captured through surveys, and associations were assessed using univariate and multivariable analyses. Pet-owning households had 1.83 times the risk (95% CI = 1.53, 2.20) of finding ticks crawling on and 1.49 times the risk (95% CI = 1.20, 1.84) of finding ticks attached to household members compared to households without pets. This large evaluation of pet ownership, human tick encounters and tickborne diseases shows that pet owners, whether of cats or dogs, are at increased risk of encountering ticks and suggests that pet owners are at an increased risk of developing tickborne disease. Pet owners should be made aware of this risk and be reminded to conduct daily tick checks of all household members, including the pets, and to consult their veterinarian regarding effective tick control products.

Tick-borne diseases, due to a diversity of bacterial pathogens, represent a significant and increasing public health threat throughout the Northern Hemisphere. A high-throughput 16S V1-V2 rRNA gene-based metagenomics assay was developed and evaluated using13,000 residual samples from patients suspected of having tick-borne illness and1,000 controls. Taxonomic predictions for tick-borne bacteria were exceptionally accurate, as independently validated by secondary testing. Overall, 881 specimens were positive for bacterial tick-borne agents. Twelve tick-borne bacterial species were detected, including two novel pathogens, representing a 100% increase in the number of tick-borne bacteria identified compared to what was possible by initial PCR testing. In three blood specimens, two tick-borne bacteria were simultaneously detected. Seven bacteria, not known to be tick transmitted, were also confirmed to be unique to samples from persons suspected of having tick-borne illness. These results indicate that 16S V1-V2 metagenomics can greatly simplify diagnosis and accelerate the discovery of bacterial tick-borne pathogens.

This article belongs to the Special Issue Ecology and Conservation of Freshwater Fishes Biodiversity., The bleak Alburnus alburnus is native to most of Europe. This cyprinid fish is a successful invader in the Iberian Peninsula. No studies exist on its foraging strategies on a large scale for this ecoregion. The aim of the present study was to compare dietary traits of invasive bleak among the main Iberian rivers and a ‘reference’ native bleak population from France. Bleak were sampled during May–June 2019 from the Iberian Rivers Ebro, Tagus, Guadiana, Segura and Guadalquivir and the River Saône (France). Diptera larvae and zooplankton were common food categories in the River Saône. Insect nymphs were more important in the River Ebro. The intake of plant material was higher in the River Tagus. Flying insects were more consumed in the River Guadiana. Nektonic insects were important in the River Guadalquivir. Detritus was a frequent food category for all populations, in terms of occurrence and mass. Dietary parameters followed a unimodal response in relation to the latitudinal gradient, with the maximum values for the Tagus and Guadiana populations. Overall, results suggest that this wide interpopulation variability will contribute to the species’ successful establishment throughout Mediterranean Europe, which poses a serious risk to its highly valuable native fish fauna, This research was partly supported by the Spanish Ministry of Education (FPU 13/03291), the Spanish Ministry of Economy and Competitiveness (projects CGL2015-69311-REDT and CGL2016-80820-R) and the Government of Catalonia (ref. 2014 SGR 484).

The Near Eastern fire salamander (Salamandra infraimmaculata) reaches its southern distribution range in Israel. Although the population structure has been analysed in central Israel and at the southern distribution limit, we lack knowledge on populations in the northern area, such as along Mount Hermon. S. infraimmaculata occurs at Mt. Hermon along an altitudinal gradient and appears to be fragmented by urban and agricultural landscape. We studied the genetic structure of four populations based on microsatellite loci and the mitochondrial D-loop to determine the genetic diversity and connectivity between populations. We observed moderate gene flow at lower parts, i.e. from Tel Dan and Nimrod Castle to Banias indicating extant but limited connectivity. Genetic diversity and gene flow declined along the altitudinal gradient at Mt. Hermon, reaching rock-bottom levels in the highest located population of Nimrod Pool. The observed isolation-by-elevation gradient might induce a higher extinction risk for the highland populations of S. infraimmaculata

Nearly all microbial communities are dynamic in time. Understanding how temporal dynamics in microbial community structure affect soil biogeochemistry and fertility are key to being able to predict the responses of the soil microbiome to environmental perturbations. Here, we explain the effects of soil spatial structure and relic DNA on the determination of microbial community fluctuations over time. We found that intensive spatial sampling was required to identify temporal effects in microbial communities because of the high degree of spatial heterogeneity in soil and that DNA from nonliving sources masks important temporal patterns. We identified groups of microbes with shared temporal responses and show that these patterns were predictable from changes in soil characteristics. These results provide insight into the environmental preferences and temporal relationships between individual microbial taxa and highlight the importance of considering relic DNA when trying to detect temporal dynamics in belowground communities., Few studies have comprehensively investigated the temporal variability in soil microbial communities despite widespread recognition that the belowground environment is dynamic. In part, this stems from the challenges associated with the high degree of spatial heterogeneity in soil microbial communities and because the presence of relic DNA (DNA from dead cells or secreted extracellular DNA) may dampen temporal signals. Here, we disentangle the relationships among spatial, temporal, and relic DNA effects on prokaryotic and fungal communities in soils collected from contrasting hillslopes in Colorado, USA. We intensively sampled plots on each hillslope over 6 months to discriminate between temporal variability, intraplot spatial heterogeneity, and relic DNA effects on the soil prokaryotic and fungal communities. We show that the intraplot spatial variability in microbial community composition was strong and independent of relic DNA effects and that these spatial patterns persisted throughout the study. When controlling for intraplot spatial variability, we identified significant temporal variability in both plots over the 6-month study. These microbial communities were more dissimilar over time after relic DNA was removed, suggesting that relic DNA hinders the detection of important temporal dynamics in belowground microbial communities. We identified microbial taxa that exhibited shared temporal responses and show that these responses were often predictable from temporal changes in soil conditions. Our findings highlight approaches that can be used to better characterize temporal shifts in soil microbial communities, information that is critical for predicting the environmental preferences of individual soil microbial taxa and identifying linkages between soil microbial community composition and belowground processes.

Waterfowl impoundments are hydrologically managed to provide food and habitat for migratory birds and have the potential to export nitrogen during drawdown. The goal of this study was to describe how nitrate, ammonium, and dissolved organic nitrogen dynamics varied under two different management schemes during storm events. This was made possible through monitoring the three forms of nitrogen at a high frequency (30 min) in a moist-soil managed (MSM) impoundment and seasonally flooded agricultural impoundment (Ag) for 17 months. Substantial differences in nitrogen dynamics between sites were observed when the sites were not flooded, while similar dynamics were observed during the winter flooding period. When the Ag site was drained, storm events mobilized nitrate (average 0.3 mg L−1; increase 0.8 mg L−1) and ammonium (average 0.4 mg L−1; increase 0.2 mg L−1). Under drained conditions in the MSM impoundment, rainfall reduced ammonium (average 0.5 mg L−1; decrease 0.1 mg L−1). Storms stimulated what appeared to be coupled nitrification-denitrification in both impoundments when flooded and ammonium concentrations were elevated. A surprising result of this work was observed at the Ag site, where elevated nitrate (0.5 mg L−1) was measured during high water levels when anoxic conditions were expected to support denitrification. Although water management schemes were found to be important for controlling nitrogen dynamics, other factors, such as carbon quality, require further research. This study demonstrates a high variability between storm events and the large influence of hydrologic management schemes on nitrogen dynamics during storm events.

Nutrient remobilization during leaf senescence nourishes the growing plant. Understanding the regulation of this process is essential for reducing our dependence on nitrogen fertilizers and increasing agricultural sustainability. Our laboratory is interested in chromatin changes that accompany the transition to leaf senescence. Previously, darker green leaves were reported for Arabidopsis thaliana hac1 mutants, defective in a gene encoding a histone acetyltransferase in the CREB‐binding protein family. Here, we show that two Arabidopsis hac1 alleles display delayed age‐related developmental senescence, but have normal dark‐induced senescence. Using a combination of ChIP‐seq for H3K9ac and RNA‐seq for gene expression, we identified 43 potential HAC1 targets during age‐related developmental senescence. Genetic analysis demonstrated that one of these potential targets, ERF022, is a positive regulator of leaf senescence. ERF022 is regulated additively by HAC1 and MED25, suggesting MED25 may recruit HAC1 to the ERF022 promoter to increase its expression in older leaves.

Nutrient remobilization during leaf senescence nourishes the growing plant. Understanding the regulation of this process is essential for reducing our dependence on nitrogen fertilizers and increasing agricultural sustainability. Our lab is interested in chromatin changes that accompany the transition to leaf senescence. Previously, darker green leaves were reported for Arabidopsis thaliana hac1 mutants, defective in a gene encoding a histone acetyltransferase in the CREB-binding protein family. Here, we show that two Arabidopsis hac1 alleles display delayed age-related developmental senescence, but have normal dark-induced senescence. Using a combination of ChIP-seq for H3K9ac and RNA-seq for gene expression, we identified 44 potential HAC1 targets during age-related developmental senescence. Genetic analysis demonstrated that one of these potential targets, ERF022, is a positive regulator of leaf senescence. ERF022 is regulated additively by HAC1 and MED25, suggesting MED25 recruits HAC1 to the ERF022 promoter to increase its expression in older leaves.

We investigated the connectivity of walleye pollock in the Gulf of Alaska (GOA) and linkages to the Bering Sea (BS) and Aleutian Island (AL) regions. We used a spatially-explicit Individual-based model (IBM) coupled to 6 years of a hydrodynamic model that simulates the early life history of walleye pollock in the GOA (eggs to age-0 juveniles). The processes modeled included growth, movement, mortality, feeding and the bioenergetics component for larvae and juveniles. Simulations were set to release particles on the 1st of the month (February to May) in fourteen historical spawning areas in the GOA up to the 1st of September each year. Model results reproduced the link between the Shelikof Strait spawning area and the Shumagin nursery region for March and April spawners, besides other Potential Nursery Areas (PNAs) found in the GOA. A prominent finding of this study was the appearance of the BS as important PNAs for several GOA spawning grounds, which is supported by a consistent flow into the BS through Unimak Pass. The simulations showed the highest density of simulated surviving pollock in the western Bering Sea (WBS) region with the lowest coefficients of variation of the whole domain. Three spawning sectors were defined, which aggregate multiple spawning areas in the eastern (EGOA), central (CGOA) and western Gulf of Alaska (WGOA). A connectivity matrix showed strong retention within the CGOA (25.9%) and EGOA (23.8%), but not in the WGOA (7.2%). Within the GOA, the highest connectivity is observed from EGOA to CGOA (57.8%) followed by the connection from CGOA to WGOA (24.3%). Overall, one of the most prominent connections was from WGOA to WBS (62.8%), followed by a connection from CGOA to WBS (29.2%). In addition, scenarios of shifting spawning locations and nursery sectors of GOA, BS and AL are explored and implications for walleye pollock stock structure hypotheses are discussed.

Biophysical individual-based models (IBMs) have been used to study aspects of early life history of marine fishes such as recruitment, connectivity of spawning and nursery areas, and marine reserve design. However, there is no consistent approach to validating the spatial outputs of these models. In this study, we hope to rectify this gap. We document additions to an existing individual-based biophysical model for Alaska walleye pollock (Gadus chalcogrammus), some simulations made with this model and methods that were used to describe and compare spatial output of the model versus field data derived from ichthyoplankton surveys in the Gulf of Alaska. We used visual methods (e.g. distributional centroids with directional ellipses), several indices (such as a Normalized Difference Index (NDI), and an Overlap Coefficient (OC), and several statistical methods: the Syrjala method, the Getis-Ord Gi* statistic, and a geostatistical method for comparing spatial indices. We assess the utility of these different methods in analyzing spatial output and comparing model output to data, and give recommendations for their appropriate use. Visual methods are useful for initial comparisons of model and data distributions. Metrics such as the NDI and OC give useful measures of co-location and overlap, but care must be taken in discretizing the fields into bins. The Getis-Ord Gi* statistic is useful to determine the patchiness of the fields. The Syrjala method is an easily implemented statistical measure of the difference between the fields, but does not give information on the details of the distributions. Finally, the geostatistical comparison of spatial indices gives good information of details of the distributions and whether they differ significantly between the model and the data. We conclude that each technique gives quite different information about the model-data distribution comparison, and that some are easy to apply and some more complex. We also give recommendations for a multistep process to validate spatial output from IBMs.

Motor behaviors such as walking or withdrawing the limb from a painful stimulus rely upon integrative multimodal sensory circuitry to generate appropriate muscle activation patterns. Both the cellular components and the molecular mechanisms that instruct the assembly of the spinal sensorimotor system are poorly understood. Here we characterize the connectivity pattern of a sub-population of lamina V inhibitory sensory relay neurons marked during development by the nuclear matrix and DNA binding factor Satb2 (ISR(Satb2)). ISR(Satb2) neurons receive inputs from multiple streams of sensory information and relay their outputs to motor command layers of the spinal cord. Deletion of the Satb2 transcription factor from ISR(Satb2) neurons perturbs their cellular position, molecular profile, and pre- and post-synaptic connectivity. These alterations are accompanied by abnormal limb hyperflexion responses to mechanical and thermal stimuli and during walking. Thus, Satb2 is a genetic determinant that mediates proper circuit development in a core sensory-to-motor spinal network.