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PREMISE OF THE STUDY Habitat fragmentation generates molecular genetic divergence among isolated populations but few studies have assessed phenotypic divergence and fitness in populations where the genetic consequences of habitat fragmentation are known. Phenotypic divergence could reflect plasticity, local adaptation, and/or genetic drift. METHODS We examined patterns and potential drivers of phenotypic divergence among 12 populations of jewelweed (Impatiens capensis Meerb.) that show strong molecular genetic signals of isolation and drift among fragmented habitats. We measured morphological and reproductive traits in both maternal plants within natural populations and their self-fertilized progeny grown together in a common garden. We also quantified environmental divergence between home sites and the common garden. KEY RESULTS Populations with less molecular genetic variation expressed less maternal phenotypic variation. Progeny in the common garden converged in phenotypes relative to their wild mothers but retained among-population differences in morphology, survival, and reproduction. Among-population phenotypic variance was 3-10x greater in home sites than in the common garden for 6 of 7 morphological traits measured. Patterns of phenotypic divergence paralleled environmental gradients in ways suggestive of adaptation. Progeny resembled their mothers less as the environmental distance between their home site and the common garden increased. CONCLUSIONS Despite strong molecular signatures of isolation and drift, phenotypic differences among these Impatiens populations appear to reflect both adaptive quantitative genetic divergence and plasticity. Quantifying the extent of local adaptation and plasticity and how these covary with molecular and phenotypic variation help us predict when populations may lose their adaptive capacity. This article is protected by copyright. All rights reserved.

Renal transplant patients are immunosuppressed and are at increased risk of opportunistic infections, including Nocardia infection. In renal transplant patients, information on the incidence and risk factors associated with nocardiosis is limited. To address the incidence and risk factors associated with nocardiosis in a large renal transplant population, we used the US Renal Data System (USRDS). Sequelae of allograft failure or rejection after infection were also examined. Demographics, clinical risk factors, Nocardia diagnosis, and allograft failure following Nocardia infection were queried in USRDS renal transplant patients using International Classification of Diseases, Ninth Revision (ICD-9) codes in billing claims and Centers for Medicare and Medicaid Services Form 2728. Generalized linear models were used to determine the risk factors associated with nocardiosis, and Cox proportional hazards models were used to examine the association of risk factors with graft failure among patients with Nocardia infection. Of 203,233 renal transplant recipients identified from 2001 to 2011, 657 (0.32%) were diagnosed with Nocardia infection. Pneumonia was the most frequent presentation (15.2%), followed by brain abscess (8.4%). Numerous factors associated with increased Nocardia infection included age >65 years (OR=2.10, 95% CI 1.71 to 2.59), history of transplant failure (OR=1.28, CI 1.02 to 1.60) or history of rejection (OR=4.83, CI 4.08 to 5.72), receipt of a deceased donor transplant (OR=1.23, CI 1.03 to 1.46), and treatment with basiliximab (OR=1.25, CI 1.00 to 1.55), cyclosporine (OR=1.30, CI 1.03 to 1.65), tacrolimus (OR=2.45, CI 2.00 to 3.00), or thymoglobulin (OR=1.89, CI 1.59 to 2.25). In patients with nocardiosis administration of antithymocyte globulin (HR=2.76), chronic obstructive pulmonary disease (HR=2.47), and presentation of Nocardia infection with brain abscess (HR=1.85) were associated with an increased risk of graft failure. This study provides new information to enhance early recognition and targeted treatment of nocardiosis in renal transplant patients.

Tisagenlecleucel is indicated for pediatric and young adult patients with relapsed/refractory (r/r) B-cell acute lymphoblastic leukemia (B-ALL) and adult patients with r/r diffuse large B-cell lymphoma (DLBCL). The tisagenlecleucel chimeric antigen receptor (CAR) contains a murine single-chain variable fragment domain; we examined the effects of humoral and cellular immune responses to tisagenlecleucel on clinical outcomes using 2 validated assays. Data were pooled from the ELIANA (registered at www.clinicaltrials.gov as #NCT02435849) and ENSIGN (#NCT02228096) trials in r/r B-ALL (N = 143) and the JULIET trial (#NCT02445248) in r/r DLBCL (N = 115). Humoral responses were determined by flow cytometric measurement of anti-murine CAR19 (mCAR19) antibodies in serum. Cellular responses were determined using T-cell production of interferon-γ in response to 2 different pools of mCAR19 peptides. Pretreatment anti-mCAR19 antibodies were detected in 81% of patients with r/r B-ALL and 94% of patients with r/r DLBCL. Posttreatment anti-mCAR19 antibodies were higher than patient-specific baseline in 42% of r/r B-ALL and 9% of r/r DLBCL patients. Pretreatment and posttreatment anti-mCAR19 antibodies did not affect tisagenlecleucel cellular kinetics, including maximum concentration and persistence (r2 < 0.05), clinical response (day-28 response, duration of response, and event-free survival), and safety. T-cell responses were consistent over time, with net responses

Preeclampsia is characterized by the development of elevated blood pressure during the second and third trimesters of pregnancy that is accompanied by end organ dysfunction. The pathogenesis of preeclampsia is multifactorial but is commonly characterized by endothelial dysfunction and the overproduction of antiangiogenic factors, including the soluble VEGF (vascular endothelial growth factor) receptor sFlt-1 (soluble Fms-like tyrosine kinase receptor 1). Previously, administration of exogenous VEGF-A, bound to a carrier protein called ELP (elastin-like polypeptide), significantly reduced free sFlt-1 levels and attenuated the hypertensive response in a rodent model of preeclampsia. However, VEGF-A administration induces multifactorial effects mediated through its direct activation of the Flk-1 receptor. In response to this, we developed a therapeutic chimera using ELP bound to VEGF-B, a VEGF isoform that binds to sFlt-1 but not to Flk-1. The purpose of this study was to evaluate the in vitro activity and pharmacological properties of ELP-VEGF-B and to test its efficacy in the reduced uterine perfusion pressure rat model of placental ischemia. ELP-VEGF-B was less potent than ELP-VEGF-A in stimulation of endothelial cell proliferation and matrix invasion, indicating that it is a weaker angiogenic driver. However, after repeated subcutaneous administration in pregnant rats, ELP-VEGF-B was maternally sequestered and reduced blood pressure when compared with saline treated animals following induction of placental ischemia (123.38±11.4 versus 139.98±10.56 mm Hg, P =0.0129). Blood pressure reduction was associated with a restoration of the angiogenic capacity of plasma from rats treated with ELP-VEGF-B. ELP-VEGF-B is a nonangiogenic, maternally sequestered protein with potential efficacy for treatment of preeclampsia.

In the Gulf of Alaska, commercially harvested fish species utilize habitats dominated by red tree corals (Primnoa pacifica) for shelter, feeding, and nurseries, but recent studies hint that environmental conditions may be interrupting the reproductive lifecycle of the corals. The North Pacific has experienced persistent and extreme thermal variability in recent years and this pattern is predicted to continue in coming decades. Recent discovery of deep-water emerged coral populations in Southeast Alaska fjords provided opportunity for detailed life-history studies and comparison to corals in managed habitats on the continental shelf. Here we show that sperm from deep colonies develops completely, but in shallow colonies, sperm development is prematurely halted, likely preventing successful production of larvae. We hypothesize that the divergence is due to differing temperature regimes presently experienced by the corals. Compared to deep populations below the thermocline, shallow populations experience much greater seasonal thermal variability and annual pulses of suspected near-lethal temperatures that appear to interrupt the production of viable gametes. The unique opportunity to comprehensively study emerged populations presently affected by thermal stress provides advance warning of the possible fate of deep corals in the Gulf of Alaska that will soon experience similar ocean conditions.

Molecular programs that underlie precursor progression in multiple myeloma are incompletely understood. Here, we report a disease spectrum-spanning, single-cell analysis of the Vκ*MYC myeloma mouse model. Using samples obtained from mice with serologically undetectable disease, we identify malignant cells as early as 30 weeks of age and show that these tumours contain subclonal copy number variations that persist throughout progression. We detect intratumoural heterogeneity driven by transcriptional variability during active disease and show that subclonal expression programs are enriched at different times throughout early disease. We then show how one subclonal program related to GCN2 stress response is progressively activated during progression in myeloma patients. Finally, we use chemical and genetic perturbation of GCN2 in vitro to support this pathway as a therapeutic target in myeloma. These findings therefore present a model of precursor progression in Vκ*MYC mice, nominate an adaptive mechanism important for myeloma survival, and highlight the need for single-cell analyses to understand the biological underpinnings of disease progression., The molecular programs that underlie progression in multiple myeloma (MM) are incompletely understood. Here the authors use a mouse model of MM and single-cell RNA-seq to define subclonal expression programs that arise during progression and that inform targeted therapeutic strategies.

Objective: Circulating progenitor cells possess immune modulatory properties and might mitigate inflammation that is characteristic of patients with coronary artery disease. We hypothesized that patients with fewer circulating progenitor cells (CPCs) will have higher inflammatory markers and worse outcomes. Approach and Results: Patients with stable coronary artery disease were enrolled in a prospective study enumerating CPCs as CD (cluster of differentiation)-34–expressing mononuclear cells (CD34+) and inflammation as levels of IL (interleukin)-6 and high-sensitivity CRP (C-reactive protein) levels. Patients were followed for 5 years for the end points of death and myocardial infarction with repeat inflammatory biomarkers measured after a median of 2 years. In the entire cohort of 392 patients, IL-6 and high-sensitivity CRP levels remained unchanged (0.3±2.4 pg/mL and 0.1±1.0 mg/L; P =0.45) after 2 years. CPC counts (log-transformed) were inversely correlated with the change in IL-6 levels (r, −0.17; P Conclusions: Reduced cardiovascular regenerative capacity is independently associated with progressive inflammation in patients with coronary artery disease that in turn is associated with poor outcomes. Graphic Abstract: A graphic abstract is available for this article.

Bovine clinical mastitis is an important problem for the dairy industry, and Staphylococcus aureus is a common mastitis-causing pathogen in many countries. Detailed knowledge on genetic variation of Staph. aureus strains within the bovine population, including changes over time, can be useful for mastitis control programs, because severity of disease and effects on milk production are at least partly strain-associated. Therefore, the major aim of this study was to compare sequence types of Staph. aureus isolated from cases of bovine clinical mastitis from 2002 to 2003 with sequence types of a more recent set of isolates collected from 2013 to 2018, using core genome multi-locus sequence typing (cgMLST). We also wanted to compare antibiotic resistance genes of isolates from the 2 sets, to identify changes that may have occurred over time in the Staph. aureus population. A total of 157 isolates of Staph. aureus, almost equally distributed between the 2 time periods, were subjected to high-throughput sequencing and cgMLST. The results showed that the most prevalent sequence types found among the 2002 to 2003 isolates belonged to the clonal complexes CC97, CC133, and CC151, and that those complexes still dominated among the isolates from 2013 to 2018. However, a population shift from CC133 to CC97 and CC151 over time was observed. Likewise, no important differences in prevalence of antibiotic resistance genes were found between the 2 sets of isolates. As expected, genes belonging to the major facilitator superfamily of transporter proteins, and multidrug and toxic compound extrusion transporters, were very common. Moreover, several genes and mutations conferring resistance to fosfomycin were present, but not in CC97 isolates. The beta-lactamase gene blaZ was found in only 3 out of 81 isolates from 2002 to 2003 and 1 out of 76 isolates in 2013 to 2018. In conclusion, the results indicate that mastitis-associated Staph. aureus strains circulating among dairy cows in Sweden exhibit a remarkable genotypic persistence over a time frame of close to 15 yr.

Datos disponibles en https://github.com/fmsabatini/sPlotOpen_Code, EU H2020 project BACI, Grant No. 640176 (...), Sabatini, F.M., Lenoir, J., Hattab, T., Arnst, E.A., Chytrý, M., Dengler, J., De Ruffray, P., Hennekens, S.M., Jandt, U., Jansen, F., Jiménez-Alfaro, B., Kattge, J., Levesley, A., Pillar, V.D., Purschke, O., Sandel, B., Sultana, F., Aavik, T., Aćić, S., Acosta, A.T.R., Agrillo, E., Alvarez, M., Apostolova, I., Arfin Khan, M.A.S., Arroyo, L., Attorre, F., Aubin, I., Banerjee, A., Bauters, M., Bergeron, Y., Bergmeier, E., Biurrun, I., Bjorkman, A.D., Bonari, G., Bondareva, V., Brunet, J., Čarni, A., Casella, L., Cayuela, L., Černý, T., Chepinoga, V., Csiky, J., Ćušterevska, R., De Bie, E., de Gasper, A.L., De Sanctis, M., Dimopoulos, P., Dolezal, J., Dziuba, T., El-Sheikh, M.A.E.-R.M., Enquist, B., Ewald, J., Fazayeli, F., Field, R., Finckh, M., Gachet, S., Galán-de-Mera, A., Garbolino, E., Gholizadeh, H., Giorgis, M., Golub, V., Alsos, I.G., Grytnes, J.-A., Guerin, G.R., Gutiérrez, A.G., Haider, S., Hatim, M.Z., Hérault, B., Hinojos Mendoza, G., Hölzel, N., Homeier, J., Hubau, W., Indreica, A., Janssen, J.A.M., Jedrzejek, B., Jentsch, A., Jürgens, N., Kącki, Z., Kapfer, J., Karger, D.N., Kavgacı, A., Kearsley, E., Kessler, M., Khanina, L., Killeen, T., Korolyuk, A., Kreft, H., Kühl, H.S., Kuzemko, A., Landucci, F., Lengyel, A., Lens, F., Lingner, D.V., Liu, H., Lysenko, T., Mahecha, M.D., Marcenò, C., Martynenko, V., Moeslund, J.E., Monteagudo Mendoza, A., Mucina, L., Müller, J.V., Munzinger, J., Naqinezhad, A., Noroozi, J., Nowak, A., Onyshchenko, V., Overbeck, G.E., Pärtel, M., Pauchard, A., Peet, R.K., Peñuelas, J., Pérez-Haase, A., Peterka, T., Petřík, P., Peyre, G., Phillips, O.L., Prokhorov, V., Rašomavičius, V., Revermann, R., Rivas-Torres, G., Rodwell, J.S., Ruprecht, E., Rūsiņa, S., Samimi, C., Schmidt, M., Schrodt, F., Shan, H., Shirokikh, P., Šibík, J., Šilc, U., Sklenář, P., Škvorc, Ž., Sparrow, B., Sperandii, M.G., Stančić, Z., Svenning, J.-C., Tang, Z., Tang, C.Q., Tsiripidis, I., Vanselow, K.A., Vásquez Martínez, R., Vassilev, K., Vélez-Martin, E., Venanzoni, R., Vibrans, A.C., Violle, C., Virtanen, R., von Wehrden, H., Wagner, V., Walker, D.A., Waller, D.M., Wang, H.-F., Wesche, K., Whitfeld, T.J.S., Willner, W., Wiser, S.K., Wohlgemuth, T., Yamalov, S., Zobel, M., Bruelheide, H.

The phylum Apicomplexa has been defined by the presence of the apical complex, a structure composed of secretory organelles and specific cytoskeletal elements. A conspicuous feature of the apical complex in many apicomplexans is the conoid, a hollow tapered barrel structure composed of tubulin fibers. In Toxoplasma gondii, the apical complex is a central site of convergence for calcium-related and lipid-mediated signaling pathways that coordinate conoid protrusion, microneme secretion, and actin polymerization, to initiate gliding motility. Through cutting-edge technologies, great progress has recently been made in discovering the structural subcomponents and proteins implicated in the biogenesis and stability of the apical complex and, in turn, these discoveries have shed new light on the function and evolution of this definitive structure.

X.D. was funded by a grant from the National Institute for Health Research (NIHR) Health Protection Research Unit in Genomics and Enabling Data (no. NIHR200892). M.A.H. was supported by grants from the Medical Research Council (nos. G1001787/1, MR/N002660/1 and MR/P007201/1) and the Economic and Social Research Council (no. ES/S000186/1). E.M.H. was supported by a UK Research and Innovation (UKRI) Fellowship (no. MR/S00291X/1). The discovery of antibiotics more than 80 years ago has led to considerable improvements in human and animal health. Although antibiotic resistance in environmental bacteria is ancient, resistance in human pathogens is thought to be a modern phenomenon that is driven by the clinical use of antibiotics1. Here we show that particular lineages of methicillin-resistant Staphylococcus aureus—a notorious human pathogen—appeared in European hedgehogs in the pre-antibiotic era. Subsequently, these lineages spread within the local hedgehog populations and between hedgehogs and secondary hosts, including livestock and humans. We also demonstrate that the hedgehog dermatophyte Trichophyton erinacei produces two β-lactam antibiotics that provide a natural selective environment in which methicillin-resistant S. aureus isolates have an advantage over susceptible isolates. Together, these results suggest that methicillin resistance emerged in the pre-antibiotic era as a co-evolutionary adaptation of S. aureus to the colonization of dermatophyte-infected hedgehogs. The evolution of clinically relevant antibiotic-resistance genes in wild animals and the connectivity of natural, agricultural and human ecosystems demonstrate that the use of a One Health approach is critical for our understanding and management of antibiotic resistance, which is one of the biggest threats to global health, food security and development. Publisher PDF

This article examines news coverage of Australia's 2018 National Apology to Victims of Institutional Child Sexual Abuse to reveal how conventional news values and practices produce racialised hiera...

SARS-CoV-2 spike mRNA vaccines1–3 mediate protection from severe disease as early as ten days after prime vaccination3, when neutralizing antibodies are hardly detectable4–6. Vaccine-induced CD8+ T cells may therefore be the main mediators of protection at this early stage7,8. The details of their induction, comparison to natural infection, and association with other arms of vaccine-induced immunity remain, however, incompletely understood. Here we show on a single-epitope level that a stable and fully functional CD8+ T cell response is vigorously mobilized one week after prime vaccination with bnt162b2, when circulating CD4+ T cells and neutralizing antibodies are still weakly detectable. Boost vaccination induced a robust expansion that generated highly differentiated effector CD8+ T cells; however, neither the functional capacity nor the memory precursor T cell pool was affected. Compared with natural infection, vaccine-induced early memory T cells exhibited similar functional capacities but a different subset distribution. Our results indicate that CD8+ T cells are important effector cells, are expanded in the early protection window after prime vaccination, precede maturation of other effector arms of vaccine-induced immunity and are stably maintained after boost vaccination.

Chromosome segregation in eukaryotes is driven by the kinetochore, a macromolecular complex that connects centromeric DNA to microtubules of the spindle apparatus. Kinetochores in well-studied model eukaryotes consist of a core set of proteins that are broadly conserved among distant eukaryotic phyla. By contrast, unicellular flagellates of the class Kinetoplastida have a unique set of 36 kinetochore components. The evolutionary origin and history of these kinetochores remain unknown. Here, we report evidence of homology between axial element components of the synaptonemal complex and three kinetoplastid kinetochore proteins KKT16-18. The synaptonemal complex is a zipper-like structure that assembles between homologous chromosomes during meiosis to promote recombination. By using sensitive homology detection protocols, we identify divergent orthologues of KKT16-18 in most eukaryotic supergroups, including experimentally established chromosomal axis components, such as Red1 and Rec10 in budding and fission yeast, ASY3-4 in plants and SYCP2-3 in vertebrates. Furthermore, we found 12 recurrent duplications within this ancient eukaryotic SYCP 2–3 gene family, providing opportunities for new functional complexes to arise, including KKT16-18 in the kinetoplastid parasite Trypanosoma brucei . We propose the kinetoplastid kinetochore system evolved by repurposing meiotic components of the chromosome synapsis and homologous recombination machinery that were already present in early eukaryotes.

The apical complex is the instrument of invasion used by apicomplexan parasites, and the conoid is a conspicuous feature of this apparatus found throughout this phylum. The conoid, however, is believed to be heavily reduced or missing from Plasmodium species and other members of the class Aconoidasida. Relatively few conoid proteins have previously been identified, making it difficult to address how conserved this feature is throughout the phylum, and whether it is genuinely missing from some major groups. Moreover, parasites such as Plasmodium species cycle through 3 invasive forms, and there is the possibility of differential presence of the conoid between these stages. We have applied spatial proteomics and high-resolution microscopy to develop a more complete molecular inventory and understanding of the organisation of conoid-associated proteins in the model apicomplexan Toxoplasma gondii. These data revealed molecular conservation of all conoid substructures throughout Apicomplexa, including Plasmodium, and even in allied Myzozoa such as Chromera and dinoflagellates. We reporter-tagged and observed the expression and location of several conoid complex proteins in the malaria model P. berghei and revealed equivalent structures in all of its zoite forms, as well as evidence of molecular differentiation between blood-stage merozoites and the ookinetes and sporozoites of the mosquito vector. Collectively, we show that the conoid is a conserved apicomplexan element at the heart of the invasion mechanisms of these highly successful and often devastating parasites., Proteomic characterisation of the invasion-related conoid structure in Toxoplasma reveals that this structure is ubiquitous in apicomplexan parasites, including the malaria parasite Plasmodium, where it was previously thought to be absent.

Forests in eastern North America are experiencing high densities of white-tailed deer (Odocoileus virginianus) and encroachment by invasive plants, both of which threaten native biodiversity. We review the literature on deer and invasive plant impacts focusing on studies that simultaneously evaluate the consequences of both. Deer have more frequent and more consistently negative effects than invasive plants. Widespread deer impacts now threaten many native plant species through much of their range. In contrast, invasive plant effects currently remain more localized and/or of smaller extent within forests. Deer impacts are also cumulative, hitting preferred plant species especially hard as they decline in density. This generates difficult-to-reverse legacy effects. Invasive plant effects, in contrast, tend to be more diffuse and may be more readily reversed. High deer populations also shift physical and chemical conditions in soils promoting “invasion cascades” involving non-native earthworms and certain introduced plants. Removing invasive plants without reducing deer populations can increase deer impacts on native species. Management should be integrated to address both deer and invasive plants. To safeguard and restore native biota when resources are limited, however, it may be most effective for managers to first reduce deer populations before investing in efforts to reduce invasive populations (except when invasions are at an early stage). We should rethink and reform traditional approaches to managing deer so that we can better integrate land vegetation with wildlife management to achieve broad public objectives. Interacting effects of high ungulate populations and invasive plants deserve further study to determine whether similar recommendations apply to other regions.

Herbivores may facilitate or impede exotic plant invasion, depending on their direct and indirect interactions with exotic plants relative to co-occurring natives. However, previous studies investigating direct effects have mostly used pairwise native-exotic comparisons with few enemies, reached conflicting conclusions, and largely overlooked indirect interactions such as apparent competition. Here, we ask whether native and exotic plants differ in their interactions with invertebrate herbivores. We manipulate and measure plant-herbivore and plant-soil biota interactions in 160 experimental mesocosm communities to test several invasion hypotheses. We find that compared with natives, exotic plants support higher herbivore diversity and biomass, and experience larger proportional biomass reductions from herbivory, regardless of whether specialist soil biota are present. Yet, exotics consistently dominate community biomass, likely due to their fast growth rates rather than strong potential to exert apparent competition on neighbors. We conclude that polyphagous invertebrate herbivores are unlikely to play significant direct or indirect roles in mediating plant invasions, especially for fast-growing exotic plants.

Deep-sea scleractinian cup corals are prominent members of Southern Ocean megabenthic communities, though little is known about their life history. This study used paraffin histology and scanning electron microscopy to characterize the reproduction of the scleractinian coral Balanophyllia malouinensis (Squires, 1961) from Burdwood Bank in the Drake Passage. Samples were taken in April and May via otter trawls, Blake trawls, and dredges on three separate cruises: one on the RV Lawrence M. Gould in 2006, and two on the RV Nathaniel B. Palmer in 2008 and 2011. B. malouinensis is gonochoric with rare hermaphroditism. All four spermatocyst stages were seen simultaneously in males; similarly, most females contained oocytes at varying stages of development, though seasonality or periodicity could not be determined without a wider range of sample dates. Average fecundity was 241 ± 184 oocytes/individual (n = 38) and not correlated to cup size. Maturing larvae were found in the mesenteries and coelenteron of females, indicating B. malouinensis broods its larvae. This study was the first to characterize the reproduction of a deep-sea Balanophyllia species and adds to a small but growing body of work seeking to understand the unique benthic communities of Burdwood Bank.

BACKGROUND The dual antigen iELISA uses two Streptococcus equi subsp equi surface protein antigens composed of N-terminal portions of SEQ2190 (Antigen A) and SeM (Antigen C). It is currently used to identify animals exposed to S. equi which have developed an immune response to the target antigens. OBJECTIVES To determine the usefulness of the dual antigen iELISA in a population of horses vaccinated with Pinnacle IN. We hypothesised that horses vaccinated for strangles with a live attenuated, non-encapsulated SeM-2 strain of S. equi, would seroconvert when tested 5 weeks later by the dual antigen iELISA. STUDY DESIGN Prospective case-control study. METHODS Three separate serum samples were obtained from 26 client-owned horses vaccinated annually with Pinnacle® IN and 26 university-owned (non-vaccinates): at annual strangles vaccination (S1), 5-week post-vaccination (S2) from vaccinates, and a third (S3) (at 10 weeks) from vaccinates who received a booster. Seropositivity was defined as an OD450 nm value ≥0.5 for one or both antigens. Mixed-effects ordered logistic regression analysis was used to identify factors associated with a suspect seropositive and seropositive value on the combined Antigen A and Antigen C iELISA. Post hoc pairwise comparisons of linear predictive margins were used to assess the differences in OD450 at a specific time between Antigens A and C. RESULTS Nineteen of 25 (76%) vaccinates were seropositive at S2 compared to 1 of 26 (4%) non-vaccinates. When adjusted for sample number, vaccinates were more likely to be seropositive or suspect than non-vaccinates (OR 14; P = .02, 95% CI 1.62-122.03). The OD450 value was significantly larger for Antigen C than Antigen A for vaccinates (P

Publisher's version (útgefin grein), Based on the discovery by the Resilience Project (Chen R. et al. Nat Biotechnol 34:531–538, 2016) of rare variants that confer resistance to Mendelian disease, and protective alleles for some complex diseases, we posited the existence of genetic variants that promote resilience to highly heritable polygenic disorders1,0 such as schizophrenia. Resilience has been traditionally viewed as a psychological construct, although our use of the term resilience refers to a different construct that directly relates to the Resilience Project, namely: heritable variation that promotes resistance to disease by reducing the penetrance of risk loci, wherein resilience and risk loci operate orthogonal to one another. In this study, we established a procedure to identify unaffected individuals with relatively high polygenic risk for schizophrenia, and contrasted them with risk-matched schizophrenia cases to generate the first known “polygenic resilience score” that represents the additive contributions to SZ resistance by variants that are distinct from risk loci. The resilience score was derived from data compiled by the Psychiatric Genomics Consortium, and replicated in three independent samples. This work establishes a generalizable framework for finding resilience variants for any complex, heritable disorder., SJG is supported by grants from the U.S. National Institutes of Health (5R01MH101519, 5R01AG054002), the Sidney R. Baer, Jr. Foundation, and NARSAD: The Brain & Behavior Research Foundation. SVF is supported by the K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen, Bergen, Norway, the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement number 602805, the European Union’s Horizon 2020 research and innovation programme under grant agreement number 667302 and NIMH grants 5R01MH101519 and U01 MH109536-01. HJE is supported by grants from the U.S. National Institutes of Health (U10 AA008401; U01 MH109532). Statistical analyses were conducted on the Genetic Cluster Computer, which is financially supported by the Netherlands Scientific Organization (NOW; 480-05-003) along with a supplement from the Dutch Brain Foundation and VU University. The Danish iPSYCH (The Lundbeck Foundation Initiative for Integrative Psychiatric Research) and GEMS2 teams acknowledge funding from The Lundbeck Foundation (grant no R102-A9118 and R155-2014-1724), the Stanley Medical Research Institute, an Advanced Grant from the European Research Council (project no: 294838), the Danish Strategic Research Council and grants from Aarhus University to the iSEQ and CIRRAU centers. The Danish National Biobank resource at Statens Serum Institut was supported by the Novo Nordisk Foundation. Computational resources for handling and statistical analysis of iPSYCH data on the GenomeDK HPC facility were provided by the iSEQ center, Aarhus University, Denmark (grant to ADB).

Transduction of primary human natural killer (NK) cells with lentiviral vectors has historically been challenging. We sought to evaluate multiple parameters to optimize lentiviral transduction of human peripheral blood NK cells being expanded to large numbers using a good manufacturing practice (GMP)-compliant protocol that utilizes irradiated lymphoblastoid (LCL) feeder cells. Although prestimulation of NK cells with interleukin (IL)-2 for 2 or more days facilitated transduction with vesicular stomatitis virus glycoprotein (VSVG)-pseudotyped lentivirus, there was a subsequent impairment in the capacity of transduced NK cells to proliferate when stimulated with LCL feeder cells. In contrast, incubation of human NK cells with LCL feeder cells plus IL-2 before transduction in the presence of the TBK1 inhibitor BX795 resulted in efficient lentiviral integration (mean of 23% transgene+ NK cells) and successful subsequent proliferation of the transduced cells. Investigation of multiple internal promoter sequences within the same lentiviral vector revealed differences in percentage and level of transgene expression per NK cell. Bicistronic lentiviral vectors encoding both GFP and proteins suitable for the isolation of transduced cells with magnetic beads led to efficient transgene expression in NK cells. The optimized approaches described herein provide a template for protocols that generate large numbers of fully functional and highly purified lentivirus-transduced NK cells for clinical trials., Graphical Abstract, By systematically evaluating multiple potential influencing factors, Childs and colleagues merge protocols to optimize lentiviral transduction and ex vivo expansion of human NK cells via co-culture with irradiated LCL feeder cells to produce large numbers of genetically modified NK cells for cancer immunotherapy.

The carapace length (CL) at which American lobster (Homarus americanus) females reach maturity can be used to evaluate egg production, growth patterns, and the overall health of lobster stocks. The female maturity datasets used to represent Gulf of Maine (GOM) lobsters in the 2015 Atlantic States Marine Fisheries Commission American Lobster Stock Assessment were collected in the 1990s by the Maine Department of Marine Resources at two coastal sites. Many studies have demonstrated an inverse relationship between temperature and the size at maturity in female lobsters, and GOM waters have warmed significantly over this period. To update these GOM maturity datasets, we used ovarian staging to determine the maturity status of over 1200 females from fives sites over 3 years. Broad application of this methodology in tandem with key growth measurements on females 50–120 mm CL allowed us to characterize reproductive development and generate maturity ogives (proportion mature at a given CL). We observed a latitudinal gradient in the size at maturity across this coastal region of the GOM and quantified a decrease in this size over 25 years. These findings have implications for future stock assessment approaches and management measures implemented to sustain this valuable fishery.

Darwin spent years investigating the effects of self-fertilization, concluding that "nature abhors perpetual self-fertilization." Given that selection purges inbred populations of strongly deleterious mutations and drift fixes mild mutations, why does inbreeding depression (ID) persist in highly inbred taxa and why do no purely selfing taxa exist? Background selection, associations and interference among loci, and drift within small inbred populations all limit selection while often increasing fixation. These mechanisms help to explain why more inbred populations in most species consistently show more fixed load. This drift load is manifest in the considerable heterosis regularly observed in between-population crosses. Such heterosis results in subsequent high ID, suggesting a mechanism by which small populations could retain variation and inbreeding load. Multiple deleterious recessive mutations linked in repulsion generate pseudo-overdominance. Many tightly linked load loci could generate a balanced segregating load high enough to sustain ID over many generations. Such pseudo-overdominance blocks (or "PODs") are more likely to occur in regions of low recombination. They should also result in clear genetic signatures including genomic hotspots of heterozygosity; distinct haplotypes supporting alleles at intermediate frequency; and high linkage disequilibrium in and around POD regions. Simulation and empirical studies tend to support these predictions. Additional simulations and comparative genomic analyses should explore POD dynamics in greater detail to resolve whether PODs exist in sufficient strength and number to account for why ID and load persist within inbred lineages.

BACKGROUND Strangles is a worldwide infectious disease caused by Streptococcus equi subsp. equi that affects the upper respiratory tract of horses. Streptococcus equi subsp. equi characterisation by seM-typing is internationally used for epidemiological studies and comparison of isolates. OBJECTIVES To identify and to compare the seM-types of Argentinian isolates of Streptococcus equi subsp. equi. STUDY DESIGN Investigation of bacterial isolates using molecular and phylogenetic approaches. METHODS A total of 59 Argentinian isolates of Streptococcus equi subsp. equi obtained between 2007 and 2019 were studied by seM-typing. The sequence similarity of Argentinian seM-types and the other alleles available on the seM database was determined using BLAST and phylogenetic analysis was performed using the Neighbour-Joining algorithm. The amino acid sequences were predicted and compared with the predicted amino acid sequence of the reference strain 4047 using the MEGA 7 software and PROVEAN tool. RESULTS Eight seM-types were found among the isolates. Only one of them (seM-61) has been previously reported and the other seven alleles (seM-129, seM-130, seM-131, seM-132, seM-133, seM-134 and seM-135) were novel seM sequences. High genetic similarity was observed among the Argentinian seM-types, with the exception of seM-130. No functional effects of amino acid differences were predicted. MAIN LIMITATIONS The number of related and unrelated isolates per year. CONCLUSIONS Seven novel seM-types and seM-61 that were previously reported in Brazil were circulating in Argentina which were identified as circulating in Argentinian horses between 2007 and 2019. The high genetic similarity among the Argentinian and Brazilian seM-types suggests that there is a geographical distribution of strain types. The geographical restriction of strains is likely to reflect the movement of horses between different equine disciplines and neighbouring countries.

The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for virus infection through the engagement of the human ACE2 protein1 and is a major antibody target. Here we show that chronic infection with SARS-CoV-2 leads to viral evolution and reduced sensitivity to neutralizing antibodies in an immunosuppressed individual treated with convalescent plasma, by generating whole-genome ultra-deep sequences for 23 time points that span 101 days and using in vitro techniques to characterize the mutations revealed by sequencing. There was little change in the overall structure of the viral population after two courses of remdesivir during the first 57 days. However, after convalescent plasma therapy, we observed large, dynamic shifts in the viral population, with the emergence of a dominant viral strain that contained a substitution (D796H) in the S2 subunit and a deletion (ΔH69/ΔV70) in the S1 N-terminal domain of the spike protein. As passively transferred serum antibodies diminished, viruses with the escape genotype were reduced in frequency, before returning during a final, unsuccessful course of convalescent plasma treatment. In vitro, the spike double mutant bearing both ΔH69/ΔV70 and D796H conferred modestly decreased sensitivity to convalescent plasma, while maintaining infectivity levels that were similar to the wild-type virus.The spike substitution mutant D796H appeared to be the main contributor to the decreased susceptibility to neutralizing antibodies, but this mutation resulted in an infectivity defect. The spike deletion mutant ΔH69/ΔV70 had a twofold higher level of infectivity than wild-type SARS-CoV-2, possibly compensating for the reduced infectivity of the D796H mutation. These data reveal strong selection on SARS-CoV-2 during convalescent plasma therapy, which is associated with the emergence of viral variants that show evidence of reduced susceptibility to neutralizing antibodies in immunosuppressed individuals.

Iron (Fe), an essential element for plant growth, is abundant in soil but with low bioavailability. Thus, plants developed specialized mechanisms to sequester the element. Beneficial microbes have recently become a favored method to promote plant growth through increased uptake of essential micronutrients, like Fe, yet little is known of their mechanisms of action. Functional mutants of the epiphytic bacterium Azospirillum brasilense, a prolific grass-root colonizer, were used to examine mechanisms for promoting iron uptake in Zea mays. Mutants included HM053, FP10, and ipdC, which have varying capacities for biological nitrogen fixation and production of the plant hormone auxin. Using radioactive iron-59 tracing and inductively coupled plasma mass spectrometry, we documented significant differences in host uptake of Fe2+/3+ correlating with mutant biological function. Radioactive carbon-11, administered to plants as 11CO2, provided insights into shifts in host usage of ‘new’ carbon resources in the presence of these beneficial microbes. Of the mutants examined, HM053 exhibited the greatest influence on host Fe uptake with increased plant allocation of 11C-resources to roots where they were transformed and exuded as 11C-acidic substrates to aid in Fe-chelation, and increased C-11 partitioning into citric acid, nicotianamine and histidine to aid in the in situ translocation of Fe once assimilated.

Optimal diet and functional response models are used to understand the evolution of primate foraging strategies. The predictions of these models can be tested by examining the geographic and seasonal variation in dietary diversity. Dietary diversity is a useful tool that allows dietary comparisons across differing sampling locations and time periods. Bonobos (Pan paniscus) are considered primarily frugivorous and consume fruits, leaves, insects, vertebrates, terrestrial herbaceous vegetation, and flowers. Frugivores, like bonobos, are valuable for examining dietary diversity and testing foraging models because they eat a variety of species and are subject to seasonal shifts in fruit availability. Frugivorous primate species thus allow for tests of how variation in dietary diversity is correlated with variation in ecological factors. We investigated measures of dietary diversity in bonobos at two research camps across field seasons within the same protected area (N’dele and Iyema) in Lomako Forest, Democratic Republic of the Congo. We compared the results of behavioral observation (1984/1985, 1991, 1995, 2014, and 2017) and fecal washing analysis (2007 and 2009) between seasons and study period using three diversity indices (Shannon’s, Simpson’s, and SW evenness). The average yearly dietary diversity indices at N’dele were Shannon’s Hʹ = 2.04, Simpson’s D = 0.82, and SW evenness = 0.88 while at Iyema, the indices were Shannon’s Hʹ = 2.02, Simpson’s D = 0.82, and SW evenness = 0.88. Behavioral observation data sets yielded significantly higher dietary diversity indices than fecal washing data sets. We found that food item (fruit, leaf, and flower) consumption was not associated with seasonal food availability for the 2017 behavioral observation data set. Shannon’s index was lower during periods when fewer bonobo dietary items were available to consume and higher when fruit was abundant. Finally, we found that optimal diet models best-explained patterns of seasonal food availability and dietary diversity. Dietary diversity is an essential factor to consider when understanding primate diets and can be a tool in understanding variation in primate diets, particularly among frugivores. Dietary diversity varies across populations of the same species and across time, and it is critical in establishing a complete understanding of how primate diets change over time.

Patterns and practices of agricultural expansion threaten the persistence of global biodiversity. Wildlife species surviving large-scale land use changes can be exposed to a suite of contaminants that may deleteriously impact their health. There is a paucity of data concerning the ecotoxicological impacts associated with the global palm oil (Elaeis guineensis) industry. We sampled wild Malay civets (Viverra tangalunga) across a patchwork landscape degraded by oil palm agriculture in Sabah, Malaysian Borneo. Using a non-lethal methodology, we quantified the levels of 13 essential and non-essential metals within the hair of this adaptable small carnivore. We robustly assessed the biological and environmental drivers of intrapopulation variation in measured levels. Metal concentrations were associated with civet age, weight, proximity to a tributary, and access to oxbow lakes. In a targeted case study, the hair metal profiles of 16 GPS-collared male civets with differing space use patterns were contrasted. Civets that entered oil palm plantations expressed elevated aluminium, cadmium, and lead, and lower mercury hair concentrations compared to civets that remained exclusively within the forest. Finally, we paired hair metal concentrations with 34 blood-based health markers to evaluate the possible sub-lethal physiological effects associated with varied hair metal levels. Our multi-facetted approach establishes these adaptable carnivores as indicator species within an extensively altered ecosystem, and provides critical and timely evidence for future studies.

Margaritifera monodonta, or the Spectaclecase Mussel, is a federally endangered freshwater mussel species that has experienced a 55% reduction in range and is currently concentrated in 3 ri...

Anthropogenic carbon emissions released into the atmosphere is driving rapid, concurrent increases in temperature and acidity across the world's oceans. Disentangling the interactive effects of warming and acidification on vulnerable life stages is important to our understanding of responses of marine species to climate change. This study evaluates the interactive effects of these stressors on the acute response of gene expression of postlarval American lobster (Homarus americanus), a species whose geographic range is warming and acidifying faster than most of the world's oceans. In the context of our experiment, we found two especially noteworthy results: First, although physiological end points have consistently been shown to be more responsive to warming in similar experimental designs, our study found gene regulation to be considerably more responsive to elevated pCO2. Furthermore, the combined effect of both stressors on gene regulation was significantly greater than either stressor alone. Using a full factorial experimental design, lobsters were raised in control and elevated pCO2 concentrations (400 ppm and 1,200 ppm) and temperatures (16°C and 19°C). A transcriptome was assembled from an identified 414,517 unique transcripts. Overall, 1,108 transcripts were differentially expressed across treatments, several of which were related to stress response and shell formation. When temperature alone was elevated (19°C), larvae downregulated genes related to cuticle development; when pCO2 alone was elevated (1,200 ppm), larvae upregulated chitinase as well as genes related to stress response and immune function. The joint effects of end‐century stressors (19°C, 1,200 ppm) resulted in the upregulation of those same genes, as well as cellulase, the downregulation of calcified cuticle proteins, and a greater upregulation of genes related to immune response and function. These results indicate that changes in gene expression in larval lobster provide a mechanism to respond to stressors resulting from a rapidly changing environment., Using nextgen transcriptomic sequencing methodologies, our study evaluates the single and joint effects of elevated temperature and CO2 on gene regulation in the postlarval stage of the American lobster. Our study shows gene regulatory response to be considerably more affected by elevated pCO2 levels than elevated temperature, contrasting what has been shown in studies of physiological end points of larval crustaceans, as well as a strong joint effect of simultaneous stressors.

An unneutered female cat of unknown age presented bloody lesions on the edematous face, and respiratory signs. Cytology and culture from the skin sample collected with fine-needle aspiration showed yeasts inside activated macrophages, and fungal growth characteristic of Histoplasma spp., which was molecularly confirmed that was Histoplasma capsulatum var. capsulatum. The cat was successfully treated with oral itraconazole (10 mg/kg/daily) for 120 days. This is the first case report of feline histoplasmosis confirmed molecularly in Brazil.

PREMISE We tested 25 classic and novel hypotheses regarding trait-origin, trait-trait, and trait-environment relationships to account for flora-wide variation in life history, habit, and especially reproductive traits using a plastid DNA phylogeny of most native (96.6%, or 1494/1547 species) and introduced (87.5%, or 690/789 species) angiosperms in Wisconsin, USA. METHODS We assembled data on life history, habit, flowering, dispersal, mating system, and occurrence across open/closed/mixed habitats across species in the state phylogeny. We used phylogenetically structured analyses to assess the strength and statistical significance of associations predicted by our models. RESULTS Introduced species are more likely to be annual herbs, occupy open habitats, have large, visually conspicuous, hermaphroditic flowers, and bear passively dispersed seeds. Among native species, hermaphroditism is associated with larger, more conspicuous flowers; monoecy is associated with small, inconspicuous flowers and passive seed dispersal; and dioecy is associated with small, inconspicuous flowers and fleshy fruits. Larger flowers with more conspicuous colors are more common in open habitats, and in understory species flowering under open (spring) canopies; fleshy fruits are more common in closed habitats. Wind pollination may help favor dioecy in open habitats. CONCLUSIONS These findings support predictions regarding how breeding systems depend on flower size, flower color, and fruit type, and how those traits depend on habitat. This study is the first to combine flora-wide phylogenies with complete trait databases and phylogenetically structured analyses to provide powerful tests of evolutionary hypotheses about reproductive traits and their variation with geographic source, each other, and environmental conditions.

Emerging data indicate that SARS-CoV-2-specific CD8+ T cells targeting different viral proteins are detectable in up to 70% of convalescent individuals. However, very little information is currently available about the abundance, phenotype, functional capacity and fate of pre-existing and induced SARS-CoV-2-specific CD8+ T cell responses during the natural course of SARS-CoV-2 infection. Here, we define a set of optimal and dominant SARS-CoV-2-specific CD8+ T cell epitopes. We also perform a high-resolution ex vivo analysis of pre-existing and induced SARS-CoV-2-specific CD8+ T cells, applying peptide-loaded major histocompatibility complex class I (pMHCI) tetramer technology. We observe rapid induction, prolonged contraction and emergence of heterogeneous and functionally competent cross-reactive and induced memory CD8+ T cell responses in cross-sectionally analyzed individuals with mild disease following SARS-CoV-2 infection and three individuals longitudinally assessed for their T cells pre- and post-SARS-CoV-2 infection. SARS-CoV-2-specific memory CD8+ T cells exhibited functional characteristics comparable to influenza-specific CD8+ T cells and were detectable in SARS-CoV-2 convalescent individuals who were seronegative for anti-SARS-CoV-2 antibodies targeting spike (S) and nucleoprotein (N). These results define cross-reactive and induced SARS-CoV-2-specific CD8+ T cell responses as potentially important determinants of immune protection in mild SARS-CoV-2 infection.

Immunity after the flu shot The seasonal flu shot is currently recommended each year because the influenza viral strains in circulation are continuously changing and because the antibody responses produced by the vaccine decline over time. In a human study of healthy volunteers, Davis et al. tracked antibody responses after flu vaccination. They investigated whether the vaccine led to the generation of antibody-secreting plasma cells in the bone marrow, a lymphoid organ that supports the survival of these cells for years. Although vaccination did generate influenza-specific cells, most were short-lived and lost within 1 year. The fact that a small number did persist over 1 year raises prospects that the longevity of flu vaccines can be improved and provides key information for the development of universal vaccines against influenza. Science , this issue p. 237

OBJECTIVE. Multiple studies suggest CT should be a primary diagnostic tool for coronavirus disease (COVID-19) because they reported sensitivities with CT far superior to that of reverse transcriptase polymerase chain reaction (RT-PCR) testing. This review aimed to assess these reports and found chest CT to have a clinical utility that is limited, particularly for patients who show no symptoms and patients who are screened early in disease progression. CONCLUSION. CT has limited sensitivity for COVID-19 and a lower specificity than RT-PCR testing, and it carries a risk of exposing providers to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Chest CT should be considered a supplemental diagnostic tool, particularly for patients who show symptoms.

Background Chinchillas, being small mammals, are subject to many of the urinary disorders that affect other companion animals. In cats and dogs, relationships between renal length and the presence of renal pathology have been well documented through the use of diagnostic imaging. Radiography and computed tomography (CT) are two commonly utilized diagnostic imaging modalities for assessing renal pathology. Presently, there are no published data on chinchilla kidney size and appearance using radiography or CT. This study aimed to determine healthy adult chinchilla renal size and correlate renal size to L2 body length and aortic diameter by use of radiography and CT. Methods Twenty-eight healthy adult chinchillas with no clinical evidence of renal disease were placed under sedation for acquisition of radiographs and CT imaging. Both kidneys were measured on the radiographic views (right lateral, left lateral and ventrodorsal), when identifiable. Renal length, L2 body length and abdominal aortic diameter were measured on CT multiplanar reformatted images in specified planes and windows. Results In 25% (7/28) of chinchillas, the renal length of either kidney could not be obtained on any of the three radiographic views due to superimposition of fluid and ingesta within the alimentary tract. Both the right and left kidney were identified on CT images in 100% of chinchillas (28/28). The left renal and right renal 95% confidence intervals from the dorsal plane CT images were 2.26 – 2.33 cm and 2.31 – 2.39 cm, respectively. No correlation was found between kidney length, L2 body length and diameter of the abdominal aorta on CT images. Conclusions Based on dorsal plane CT imaging, normal chinchilla renal length is proposed to be between 2.25–2.4 cm. Clinical Relevance Radiographic evaluation of chinchilla renal length is greatly limited due to their large fluid- and ingesta-filled colon and cecum, as is typical of animals that are hindgut fermenters. Through establishment of a normal CT reference interval for renal length and the ever-increasing clinical utility of CT, clinicians can better evaluate chinchilla renal length and identify alterations that may indicate pathologic processes.

Nest design and characteristics can influence the microclimatic conditions in the nest. Nest-dwelling ectoparasites are sensitive to temperature and moisture and as such the conditions in the nest can influence parasite infestations. The endangered African penguin (Spheniscus demersus) breeds in different nest types and as yet little is known with regard to the microclimate and parasite infestation within these nests. This study characterized the microclimatic conditions in natural open, natural covered (with vegetation) and artificial nests, and assessed the relationship between nest characteristics (type, age, distance from the coast, orientation and entrance opening) and in-nest ectoparasite infestations and the health of African penguins in Stony Point, South Africa. Penguins (50 adults and 192 chicks) and their nests (n = 308) were sampled in 2016 and 2017. Soil temperature was higher in artificial than in natural nests, and soil and nest material moisture was lower in artificial and natural covered nests than natural open. Ectoparasite infestations were higher under warmer and drier conditions, in artificial nests and nests near the coastline. Penguin (adult and chick) body mass and chick body condition were lower in warmer nests and total plasma protein (in adults and checks) was lower in drier nests. Given the potential adverse effects of ectoparasites on host species, it is recommended that conservation agencies implement a monitoring programme to assess the ectoparasite infestation in artificial nests across multiple colonies. This information will facilitate a more holistic penguin conservation management plan that may prevent further detrimental effects on this endangered penguin species.

Carbon dioxide (CO2 ) has been approved by the US Environmental Protection Agency as a new aquatic pesticide to control invasive Asian carps and other aquatic nuisance species in the United States. However, limited CO2 toxicity data could make it challenging for resource managers to characterize the potential risk to nontarget species during CO2 applications. The present study quantified the toxicity of CO2 to 2 native riverine fishes, bluegill (Lepomis macrochirus) and fathead minnow (Pimephales promelas), using 12-h continuous flow-through CO2 exposure at 5, 15, and 25 °C water temperatures. Resulting survival indicated that bluegill (median lethal concentration [LC50] range 91-140 mg/L CO2 ) were more sensitive to CO2 than fathead minnow (LC50 range 235-306 mg/L CO2 ) across all water temperatures. Bluegill were also more sensitive to CO2 at 5 °C (LC50 91 mg/L CO2 , 95% CI 85-96 mg/L CO2 ) than at 25 °C (LC50 140 mg/L CO2 , 95% CI 135-146 mg/L CO2 ). Fathead minnow showed an opposite response and were less sensitive at 5 °C (LC50 306 mg/L CO2 , 95% CI 286-327 mg/L CO2 ) relative to 25 °C (LC50 235 mg/L CO2 , 95% CI 224-246 mg/L CO2 ). Our results show that CO2 toxicity can differ by species and water temperature. Data from the present study may inform decisions related to the use of CO2 as a control tool. Environ Toxicol Chem 2020;39:2247-2255. Published 2020. This article is a U.S. government work and is in the public domain in the USA.

Extant land plants consist of two deeply divergent groups, tracheophytes and bryophytes, which shared a common ancestor some 500 million years ago. While information about vascular plants and the two of the three lineages of bryophytes, the mosses and liverworts, is steadily accumulating, the biology of hornworts remains poorly explored. Yet, as the sister group to liverworts and mosses, hornworts are critical in understanding the evolution of key land plant traits. Until recently, there was no hornwort model species amenable to systematic experimental investigation, which hampered detailed insight into the molecular biology and genetics of this unique group of land plants. The emerging hornwort model species, Anthoceros agrestis, is instrumental in our efforts to better understand not only hornwort biology but also fundamental questions of land plant evolution. To this end, here we provide an overview of hornwort biology and current research on the model plant A. agrestis to highlight its potential in answering key questions of land plant biology and evolution.

A 1.47 Å resolution crystal structure of human inositol monophosphatase bound to the inhibitor ebselen is presented. In the structure, ebselen forms a selenosulfide bond to Cys141 and ebselen-mediated contacts between two dimers give a tetramer with approximate 222 symmetry., Inositol monophosphatase (IMPase) is inhibited by lithium, which is the most efficacious treatment for bipolar disorder. Several therapies have been approved, or are going through clinical trials, aimed at the replacement of lithium in the treatment of bipolar disorder. One candidate small molecule is ebselen, a selenium-containing antioxidant, which has been demonstrated to produce lithium-like effects both in a murine model and in clinical trials. Here, the crystallization and the first structure of human IMPase covalently complexed with ebselen, a 1.47 Å resolution crystal structure (PDB entry 6zk0), are presented. In the complex with human IMPase, ebselen in a ring-opened conformation is covalently attached to Cys141, a residue located away from the active site. IMPase is a dimeric enzyme and in the crystal structure two adjacent dimers share four ebselen molecules, creating a tetramer with approximate 222 symmetry. In the crystal structure presented in this publication, the active site in the tetramer is still accessible, suggesting that ebselen may function as an allosteric inhibitor or may block the binding of partner proteins.

The aim of this study was to evaluate the effect of sampling technique and milk fraction on bovine milk microbiota data and to compare the microbiota in milk to microbiota on the teat end and in the teat canal. Representative milk samples are highly important for assessment of bacteriological findings and microbiota in milk. Samples were obtained from 5 healthy lactating dairy cows at udder quarter level during 1 milking. Swab samples from the teat. end and teat canal, and milk samples collected using different techniques and in different milk fractions were included. Milk was collected by hand stripping and through a teat canal cannula before and after machine milking, through a trans-teat wall needle aspirate after milking, and from udder quarter composite milk. The microbiota of the samples was analyzed with sequencing of the V1-V3 region of the 16S rRNA gene. In addition, somatic cell counts and bacterial cultivability were analyzed in the milk samples. Microbiota data were analyzed using multivariate methods, and differences between samples were tested using analysis of similarity (ANOSIM). Differences between samples were further explored via individual studies of the 10 most abundant genera. The microbiota on the teat end, in the teat canal, and in udder quarter composite milk, collected using a milking machine, differed in composition from the microbiota in milk collected directly from the udder quarter. No differences in milk microbiota composition were detected between hand-stripped milk samples, milk samples taken through a teat canal cannula, or milk samples taken as a trans-teat wall needle aspirate before or after milking. We conclude that for aseptic milk samples collected directly from the lactating udder quarter, sampling technique or milk fraction has minor effect on the microbiota composition.

Floral extracts (FEs) can influence the infectivity and epidemiology of fruit infecting Colletotrichum species. In this study, Colletotrichum fioriniae responded to cranberry FEs with an increased rate and magnitude of secondary conidiation and appressorium formation. Four other cranberry fruit rotting species also showed an increased rate of germination in the presence of FEs. However, increased appressorium formation was observed only in the latent pathogens Coleophoma cylindrospora, Colletotrichum fructivorum, and Colletotrichum fioriniae. Two other fruit rotting species, Phyllosticta vaccinii and Allantophomopsis lycopodina, did not form appressoria while secondary conidiation was only seen with the Colletotrichum spp. When conidia of Colletotrichum fioriniae were inoculated in the presence of FE, the incidence of disease was greater on cranberry fruit. Conidia of this species also formed appressoria at lower than expected temperatures in the presence of FE. Dissection of the flowers revealed that the corolla (with stamens and stigma) was the most stimulatory part of the inflorescence. These observations suggest an important and ephemeral role of flowers in the epidemiology of fruit rot. Stimulatory floral signals were readily detected using a conidial germination bioassay and rainwater samples collected from the plant canopy throughout the growing season confirmed that bioactivity was highest during the bloom period, and declined as the fruit developed. The data presented show that floral signals can alter the growth patterns of a larger than previously observed range of fungi and the mobility of floral signals within the canopy implicates these phenology-specific cues in modifying the disease cycles of numerous plant pathogens.

Zebra mussels (Dreissena polymorpha) have exacerbated the decline of native freshwater mussels (order Unionida) in North America since their arrival in the 1980s. Options for controlling invasive mussels, particularly in unionid mussel habitats, are limited. Previously, carbon dioxide (CO2) showed selective toxicity for zebra mussels, relative to unionids, when applied in cool water (12 °C). We first determined 96‐h lethal concentrations of CO2 at 5 and 20 °C to zebra mussels and responses of juvenile plain pocketbook (Lampsilis cardium). Next, we compared the time to lethality for zebra mussels at 5, 12, and 20 °C during exposure to partial pressure of CO2 (PCO2) values of 110 to 120 atm (1 atm = 101.325 kPa) and responses of juvenile plain pocketbook and fragile papershell (Leptodea fragilis). We found efficacious CO2 treatment regimens at each temperature that were minimally lethal to unionids. At 5 °C, plain pocketbook survived 96‐h exposure to the highest PCO2 treatment (139 atm). At 20 °C, the 96‐h lethal concentration to 10% of animals (LC10) for plain pocketbook (173 atm PCO2, 95% CI 147–198 atm) was higher than the LC99 for zebra mussels (118 atm PCO2, 95% CI 109–127 atm). Lethal time to 99% mortality (LT99) of zebra mussels in 110 to 120 atm PCO2 ranged from 100 h at 20 °C to 300 h at 5 °C. Mean survival of both plain pocketbook and fragile papershell juveniles exceeded 85% in LT99 CO2 treatments at all temperatures. Short‐term infusion of 100 to 200 atm PCO2 at a range of water temperatures could reduce biofouling by zebra mussels with limited adverse effects on unionid mussels. Environ Toxicol Chem 2020;39:1546–1557. Published 2020. This article is a U.S. Government work and is in the public domain in the USA. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Indirect interactions mediated by shared enemies or mutualists (i.e. apparent competition) can influence whether invasive plants harm or benefit co‐occurring species. However, studies to date have largely examined single pairwise interactions, limiting our understanding of the interplay among different types of interactions and whether indirect impacts systematically favour native or exotic species. Predicting indirect interaction strength has also proven challenging, and it remains unclear whether the strengths of different indirect interactions are correlated. We conducted a field experiment in a grassland invaded by Scotch broom Cytisus scoparius to compare the strength of its indirect impacts, via both soil fungi and herbivores, on 21 native and exotic legume species growing in pots buried in the ground. Direct interactions of plants with soil fungi were controlled using nylon mesh pot windows of differing porosity (1 or 38 µm) to prevent or allow soil fungi hyphal growth. Arthropod herbivores were controlled through spraying pyrethrum pesticide. To assess indirect impacts, interactions were compared between plants adjacent to or 50 m away from an extensive Scotch broom invasion. We measured plant performance (survival, height and biomass), arthropod and hare herbivory, and rhizobia nodulation. Despite increasing arthropod herbivory of both native and exotic plant species, Scotch broom had a net positive impact on their survival and growth, through sheltering them from abiotic stress, and indirectly via beneficial soil fungi and release from hare browsing. Soil fungi also increased arthropod herbivory, decreased rhizobia nodulation and disproportionately promoted the growth of exotic plants. Overall, exotic plants experienced stronger interactions, which favoured them with beneficial soil fungi and rhizobia but not hare browsing. Finally, indirect interaction strength was not correlated among indirect interactions mediated by different interaction partners. Synthesis. We demonstrate that invaders affect their competitors through multiple interacting indirect pathways that were stronger than direct ‘nurse plant’ effects, emphasizing the importance of a community‐level approach to studying biological invasions. Exotic species experienced stronger positive and negative impacts than natives, but were facilitated overall, potentially contributing to exotic dominance in communities.

Darwin observed that form, and in his view, meaning, of facial behaviour (observable changes in the appearance of the face, often termed facial ‘expression’) is similar between a wide range of species and concluded that this must be due to a shared ancestral origin. Yet, as with all social behaviours, exactly how to define similarity and determine homology is debated. Facial behaviour is linked to specific facial muscle movements, so one important factor in determining homology is the anatomical basis of facial behaviours that appear similar in both appearance and social function. The Facial Action Coding System (FACS) was developed for the scientific measurement of human facial behaviour and is based on individual facial muscle movements (Ekman and Friesen, 1978). FACS has since been modified for use with various non-human primate species (chimpanzees, macaques, hylobatids, orangutans) and domestic species (dogs, cats, horses). These FACS can be used to trace continuity of form in facial behaviour across species and build a better understanding of the evolution of facial communication in mammals.