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The manuscript discusses how to incorporate random effects for quantile regression models for clustered data with focus on settings with many but small clusters. The paper has three contributions: (i) documenting that existing methods may lead to severely biased estimators for fixed effects parameters; (ii) proposing a new two-step estimation methodology where predictions of the random effects are first computed {by a pseudo likelihood approach (the LQMM method)} and then used as offsets in standard quantile regression; (iii) proposing a novel bootstrap sampling procedure in order to reduce bias of the two-step estimator and compute confidence intervals. The proposed estimation and associated inference is assessed numerically through rigorous simulation studies and applied to an AIDS Clinical Trial Group (ACTG) study., Accepted for Econometrics and Statistics

Purpose: We investigated whether hepcidin and erythroferrone (ERFE) could complement the Athlete Biological Passport (ABP) in indirectly detecting a 130 mL packed red blood cells (RBCs) autologous blood transfusion. Endurance performance was evaluated.Methods: Forty-eight healthy men (n = 24) and women (n = 24) participated. Baseline samples were collected weekly followed by randomization to a blood transfusion (BT, n = 24) or control group (CON, n = 24). Only the BT group donated 450 mL whole blood from which 130 mL RBCs was reinfused four weeks later. Blood samples were collected 3, 7, 14, 21 and 28 days after donation, and 3, 6, and 24 hours and 2, 3, and 6 days following reinfusion. In the CON group samples were collected with the same frequency. Endurance performance was evaluated by a 650-kCal time trial (n = 13) before and one and six days after reinfusion.Results: A time×treatment effect existed (P < 0.05) for hepcidin and ERFE. Hepcidin was increased (P < 0.01) ~110 and 89% six and 24 hours after reinfusion. Using an individual approach (99% specificity, e.g. allowing 1:100 false-positive), sensitivities, i.e. true positives, of 30% and 61% was found for hepcidin and ERFE, respectively. For the ABP, the most sensitive marker was Off-hr score ([Hb] (g·L-1) - 60 × √RET%) (P < 0.05) with a maximal sensitivity of ~58% and ~ 9% following donation and reinfusion, respectively. Combining the findings for hepcidin, ERFE and the ABP yielded a sensitivity across all time-points of 83% following reinfusion in BT. Endurance performance increased 24 hours (+6.4%, P < 0.01) and six days following reinfusion (+5.8%, P < 0.01).Conclusions: Hepcidin and ERFE may serve as biomarkers in an anti-doping context following an ergogenic, small-volume blood transfusion.

Alcohol consumption increases circulating high-density lipoprotein cholesterol (HDL-C), but HDL protein cargo may better reflect HDL function. This study examined the associations between alcohol intake and HDL subspecies containing or lacking apoC3, apoE, and apoJ in a well-phenotyped cohort. We performed a cross-sectional analysis of 2092 Cardiovascular Health Study participants aged 70 or older with HDL subspecies measured in stored specimens from 1998 to 1999. Associations between alcohol intake and apoA1 defined HDL subspecies lacking or containing apoC3, apoE, and apoJ, and circulating levels of total apoA1, apoC3, apoE, and apoJ were examined. HDL subspecies lacking and containing apoC3, apoE, and apoJ were all positively associated with alcohol intake, with ∼1% per additional drink per week or ∼7% per additional drink per day (subspecies without the apolipoproteins, P ≤ 2 × 10−9, subspecies with the apolipoproteins, P ≤ 3 × 10−5). Total apoA1 was also directly associated with alcohol consumption, with a 1% increase per additional drink per week (P = 1 × 10−14). Total apoC3 blood levels were 0.5% higher per additional drink per week (P = 0.01), but the association was driven by a few heavily drinking men. Alcohol intake was positively associated with HDL subspecies lacking and containing apoC3, apoE, or apoJ, and with total plasma apoA1. ApoC3 was directly, albeit not as robustly associated with alcohol intake. HDL protein cargo is crucial for its anti-atherosclerotic functions, but it remains to be determined whether HDL subspecies play a role in the putative association between limited alcohol intake and lower risk of coronary heart disease.

Over-fertilization has caused significant phosphorus (P) accumulation in Chinese greenhouse vegetable production (GVP) soils. This study, for the first time, quantified profile P accumulation directly from soil P measurements, as well as subsoil P immobilization, in three alkaline coarse-textured GVP soil profiles with 5 (S5), 15 (S15), and 30 (S30) years of cultivation in Tongshan, Southeast China. For each profile, soil samples were collected at depths of 0–10 (topsoil), 10–20, 20–40, 40–60, 60–80, and 80–100 cm. Phosphorus accumulation was estimated from the difference in P contents between topsoil and parent material (60–100 cm subsoil). Phosphorus mobility was assessed from measurements of water-soluble P concentration (PSol). Finally, P sorption isotherms were produced using a batch sorption experiment and fitted using a modified Langmuir model. High total P contents of 1 980 (S5), 3 190 (S15), and 2 330 (S30) mg kg−1 were measured in the topsoils versus lower total P content of approximately 600 mg kg−1 in the 80–100 cm subsoils. Likewise, topsoil PSol values were very high, varying from 6.4 to 17.0 mg L−1. The estimated annual P accumulations in the topsoils were 397 (S5), 212 (S15), and 78 (S30) kg ha−1 year−1. Sorption isotherms demonstrated the dominance of P desorption in highly P-saturated topsoils, whereas the amount of adsorbed P increased in the 80–100 cm subsoils with slightly larger P adsorption capacity. The total P adsorption capacity of the 80–100 cm subsoils at a solution P concentration of 0.5 mg L−1 was 15.7 (S5), 8.7 (S15), and 6.5 (S30) kg ha−1, demonstrating that subsoils were unable to secure P concentrations in leaching water below 0.5 mg L−1 because of their insufficient P-binding capacity.

We investigated whether immature reticulocyte fraction (IRF) and immature reticulocytes to red blood cells ratio (IR/RBC) are sensitive biomarkers for low-dose recombinant human erythropoietin (rhEpo) treatment at sea-level (SL) and moderate altitude (AL) and whether multi (FACS) or single (Sysmex-XN) fluorescence flow cytometry is superior for IRF and IR/RBC determination. Thirty-nine participants completed two interventions, each containing a four-week baseline, a four-week SL or AL (2,230m) exposure and a four-week follow-up. During exposure, rhEpo (20 IU·kg-1) or placebo (PLA) was injected at SL (SLrhEpo n = 25, SLPLA n = 9) and AL (ALrhEpo n = 12, ALPLA n = 27) every second day for three weeks. Venous blood was collected weekly. Sysmex measurements revealed that IRF and IR/RBC was up to ~70% (P < 0.01) and ~190% (P < 0.001) higher in SLrhEpo than SLPLA during treatment and up to ~45% (P < 0.001) and ~55% (P < 0.01) lower post-treatment, respectively. Compared with ALPLA, IRF and IR/RBC was up to ~20% (P < 0.05) and ~45% (P < 0.001) lower post-treatment in SLrhEpo, respectively. In ALrhEpo, IRF and IR/RBC was up to ~40% (P < 0.05) and ~110% (P < 0.001) higher during treatment and up to ~25% (P < 0.05) and ~40% (P < 0.05) lower post-treatment, respectively, compared with ALPLA. Calculated thresholds provided ~90% sensitivity for both biomarkers at SL and 33% (IRF) and 66% (IR/RBC) at AL. Specificity was >99%. Single-fluorescence flow cytometry coefficient of variation was >twofold higher at baseline (P < 0.001), and provided larger or similar changes compared to multi-fluorescence, albeit with smaller precision. In conclusion, IRF and IR/RBC were sensitive and specific biomarkers for low-dose rhEpo misuse at SL and AL.

The growing global population and rising demand for meat increasingly pressures the world's resources. Edible insects are a promising alternative protein source to unsustainable conventional meat. Despite this, disgust and neophobia are cited as significant barriers to the adoption of these novel foods in Western diets. The primary aim of this study was to assess the effects of providing three types of information — the taste, health, and sustainability benefits of entomophagy (i.e. the practice of eating insects) — on the willingness to try and hedonic response to insect-based foods among children. In addition, the differences between insects (buffalo worms and cricket) in unprocessed form and in various food applications were examined. Food disgust sensitivity, food neophobia, willingness to try, familiarity, and hedonic response to insect foods were measured. The implications of the appropriateness (as a food ingredient and to be raised as livestock) of two different insect species on acceptance were also explored. The data were collected through an online questionnaire administered in school classrooms from a sample of Danish children (n = 181). Results showed that communicating information about the benefits of entomophagy did not increase the willingness to try insect foods, irrespective of the type of information. Food neophobia was found to be a strong predictor of willingness to try insect foods, whereas food disgust sensitivity had no effect. There was no correlation between food disgust and food neophobia scores. Furthermore, certain types of insect products were found to be better liked than others (e.g. cookies over falafel). There was a species effect on hedonic response when presented as a whole insect although not when presented as processed products made with insect flour.

Apple production across Europe is experiencing major yield losses due to pest damage, and the need for alternative production methods are growing. Ecological infrastructures are important to create resilient production systems and increase functional biodiversity. This study investigated perennial flower strips as a means of promoting natural enemies, thereby reducing infestation by the rosy apple aphid, Dysaphis plantaginea, and fruit damage caused by it. In 2016 and 2017, effects of perennial flower strips were assessed in eight organic apple orchards with and without flower strips. In orchards with flower strips, assessments were done in tree rows at two distances: 1st and 3rd row from the flower strip. Assessments included field samples of arthropods, visual observations of D. plantaginea symptoms and observations of predators, observations of predators directly associated with aphid colonies, and fruit damage. In 2016, significantly more symptoms of D. plantaginea were observed in the 3rd row from the flower strip compared to the 1st row from the flower strip. Both years, more predators were observed in the 1st row from the flower strip than in the 3rd row from the flower strip. In 2017, more predators were observed in orchards with a flower strip than in orchards with no flower strip, and more predators were observed inside the aphid colonies in trees in the 1st row from the flower strip than in the 3rd row from the flower strip. Predator diversity in apple trees was also positively affected by proximity to a flower strip. In conclusion, perennial flower strips consistently promote the occurrence of natural enemies in their proximity, while the effect decreases with distance. The correlation between aphid abundance and predators was not always consistent, likely because the majority of the observed predators were generalists and therefore contributing to the natural regulation of the entire herbivore complex in the apple orchards.

Serial correlation and overdispersion must be handled properly in analyses of time series of counts, and parameter-driven models combine an underlying latent process with a conditional log-linear Poisson model (given the latent process) for that purpose. Regression coefficients have direct interpretations, but likelihood inference is not straight-forward. We consider a two-step procedure for estimation: First regression parameters are estimated from the marginal distribution; second parameters concerning the latent process are estimated with composite likelihood methods, based on low-order simultaneous or conditional distributions. Confidence intervals are computed by bootstrap. Properties of estimators are examined and compared to other methods in three simulation studies, and the methods are applied to two datasets from the literature concerning hospital admission related to asthma and traffic deaths.

Extensive oil palm plantations have often resulted in the decrease of soil organic carbon (SOC). Several options exist to counter this, such as recycling empty fruit bunches (EFB) as a soil amendment. However, the extent to which EFB increase SOC has been disputed. Since EFB could also be used as a climate change mitigation tool, it is necessary to truly understand their impact on SOC. The investigation of the impact of nine EFB treatments (differing in frequency and application rates) on a 27-year-old large-scale experiment (Lampung, Sumatra, Indonesia) revealed that, while EFB impacts are heterogeneous throughout the plantation, they can positively affect total SOC and permanganate oxidisable carbon (POX-C) both at shallow and deep depths (measured up to 100 cm). POX-C was closely correlated to SOC, but showed significant increases compared to the untreated control in all treatments, while total SOC was only increased in a few treatments with small and frequent rates of EFB application. Overall, between 12 (±16) and 56 (±12) t ha−1 of carbon were sequestered under the harvesting path after 21 years. Focussing on the mineral nutrition value of the EFB, oil palm companies apply a rate of 60 t of EFB every second year for their commercial production, and the analysis of three commercial plots showed that the commercial rate only increased POX-C while it had no effect on the total SOC and SOC stocks. It seems obvious that a change of paradigm is necessary to consider EFB recycling as a new management perspective, where the potential for carbon sequestration becomes an important variable for climate change mitigation besides the initial objective of integrating EFB application into the fertiliser management plan of a plantation.

Many species of the ciliate genus Strombidium can acquire functional chloroplasts from a wide range of algal prey and are thus classified as generalist non-constitutive mixotrophs. Little, however, is known about the influence of irradiance and prey availability on their ability to exploit the photosynthetic potential of the chloroplasts, and how this may explain their spatial and temporal distribution in nature. In this study, inorganic carbon uptake, growth, and ingestion rates were measured for S. cf. basimorphum under three different irradiances (10, 40, and 120 μmol photons m–2 s–1) when acclimated to three different prey densities (5 × 103, 1 × 104, and 4 × 104 cells mL–1), as well as when allowed to deplete the prey. After prey depletion, cultures survived without prey longest (∼6 days) at the medium irradiance treatment (40 μmol photons m–2 s–1), while ciliate density, inorganic carbon uptake rates, and cellular chl-a content declined fastest at the highest irradiance treatment. This indicates that the ciliates may be unable to maintain the chloroplasts functionally without replacement at high irradiances. Ingestion rates were not shown to be significantly influenced by irradiance. The maximum gross growth efficiency (GGE) in this study (1.1) was measured in cultures exposed to the medium test irradiance and lowest prey density treatment (5 × 103 cells mL–1). The relative contribution of inorganic carbon uptake to the ciliate carbon budget was also highest in this treatment (42%). A secondary GGE peak (0.99) occurred when cultures were exposed to the highest test irradiance and the medium prey density. These and other results suggest that S. cf. basimorphum, and other generalist non-constitutive mixotrophs, can flexibly exploit many different environmental conditions across the globe.

Fungi living inside plants affect many aspects of plant health, but little is known about how plant genotype influences the fungal endophytic microbiome. However, a deeper understanding of interactions between plant genotype and biotic and abiotic environment in shaping the plant microbiome is of significance for modern agriculture, with implications for disease management, breeding and the development of biocontrol agents. For this purpose, we analysed the fungal wheat microbiome from seed to plant to seeds and studied how different potential sources of inoculum contributed to shaping of the microbiome. We conducted a large-scale pot experiment with related wheat cultivars over one growth-season in two environments (indoors and outdoors) to disentangle the effects of host genotype, abiotic environment (temperature, humidity, precipitation) and fungi present in the seed stock, air and soil on the succession of the endophytic fungal communities in roots, flag leaves and seeds at harvest. The communities were studied with ITS1 metabarcoding and environmental climate factors were monitored during the experimental period. Host genotype, tissue type and abiotic factors influenced fungal communities significantly. The effect of host genotype was mostly limited to leaves and roots, and was location-independent. While there was a clear effect of plant genotype, the relatedness between cultivars was not reflected in the microbiome. For the phyllosphere microbiome, location-dependent weather conditions factors largely explained differences in abundance, diversity, and presence of genera containing pathogens, whereas the root communities were less affected by abiotic factors. Our findings suggest that airborne fungi are the primary inoculum source for fungal communities in aerial plant parts whereas vertical transmission is likely to be insignificant. In summary, our study demonstrates that host genotype, environment and presence of fungi in the environment shape the endophytic fungal community in wheat over a growing season.

Background: Fungi living inside seeds, leaves and roots of plants affect many aspects of plant health. Understanding the role of plant genotype, presence of fungal inoculum in the environment, abiotic environmental factors, and interaction of those factors in shaping the plant-associated fungal microbiome is of great relevance for modern agriculture. This knowledge can have direct implications in plant disease management, plant breeding, and development of microorganism-based biocontrol agents. These are important tools also in the context of increasing sustainability and adaption of crop production to a changing climate. In order to disentangle the effects of host genotype, environmental factors, and fungi present in seed stock, air and soil on endophytic fungal communities over one generation, we conducted a large-scale pot experiment with closely related cultivars of wheat over one growth-season. We studied fungi present inside the plants (endophytic fungi), in soil and air with metabarcoding, and monitored abiotic factors during the experimental period.Results: Abiotic environmental factors, wheat genotype, and wheat tissue type were all found to influence fungal communities significantly. While the effect of wheat genotype was limited, there was evidence for host genetic control of fungal communities in leaves and roots but not in seeds. The degree of relatedness between wheat cultivars and resistance levels to the leaf disease STB was not reflected in the microbiome. The effect of host genetic control on the fungal community did not differ between abiotic environments. For the phyllosphere, abiotic environmental factors largely explained differences in fungal community abundance, diversity, and presence of pathogens between the locations, whereas location-dependent differences affected the rhizosphere communities to an only small extent. We found indications that airborne fungi are the primary source of inoculum for above-ground fungal communities of wheat, since a large proportion of leaf-associated fungi were also present in the air but not in soil and seed stocks.Conclusions: Our study demonstrates how the factors genotype, environment, and presence of fungi in the environment shape the endophytic fungal community in wheat over one growing season.

The liquid fraction after liquid/solid separation of biogas digestate has a high potential as a fertilizer due to its high nutrient concentration. However, the direct application of digestate in agricultural fields results in practical problems due to its voluminous nature. One solution to this could be to concentrate nutrients onto sorbents such as biochar or zeolites, which can subsequently be used as a fertilizer. This study investigated the ability of biochar and zeolite ‘clinoptilolite' enriched with digestate nutrients to supply nitrogen (N) when used as a fertilizer. A pot experiment with ryegrass was conducted to test the effect of a nutrient-enriched biochar and clinoptilolite by determining plant biomass growth and N uptake. This included untreated biochar and clinoptilolite as controls and two levels of N application (15 and 45 mg N per pot) each at two initial loading ratios (low and high). Nutrient-enriched biochar and clinoptilolite increased plant biomass yield (up to 1.02 and 2.39 g per pot) and N uptake (up to 11.23 and 39.94 mg N per pot) compared to the untreated sorbents treatments. Initial loading ratio had a significant effect on plant biomass response and apparent N recovery (ANR) for enriched clinoptilolite, and lower initial loading ratio improved plant growth. In contrast to clinoptilolite, higher initial loading ratio resulted in higher ANR. In conclusion, our results reveal that N released from enriched clinoptilolite and biochar could be taken up by the plants, clinoptilolite performed more effectively than biochar, and initial loading ratio affected the performance of the sorbents when used as a fertilizer.

A pathogenicity assay for in planta screening of biological control agents (BCAs) towards sheath blight in rice was developed, and used to evaluate a panel of Rhizoctonia solani Kuhn hyphae-associated bacteria for their ability to control sheath blight under screenhouse conditions. The best performing BCA, Burkholderia sp. strain A-7.3, was selected for field trials. Disease incidence and disease severity were significantly reduced leading to a significant grain yield increase of 7%. Furthermore, R. solani sclerotia formation and sclerotia viability were significantly reduced, thus lowering the reservoir for primary infections in the next crop cycle. The paper presents a reliable pathogenicity assay that can be used to test BCA performance under different scenarios e.g. plant variety and soil conditions, and help predict translation of results obtained under screenhouse conditions to field performance. Furthermore, it supports the hypothesis that hyphae-associated bacteria are a promising source of niche-specific BCAs towards fungal pathogens.

Intensive phosphorus (P) inputs to plastic-covered greenhouse vegetable production (PGVP) in China has led to excessive soil P accumulation increasing the potential for leaching to surface waters. This study examined the mobility and hence the potential risk of P losses through correlations between soil solution P (PSol) and soil extractable P as determined by conventional soil P test methods (STPs) including degree of P saturations (DPSs), and diffusive gradient in thin-films (DGT P) technique. A total of 75 topsoil samples were chosen from five representative Chinese PGVPs covering a wide range of physiochemical soil properties and cultivation history. Total P and Olsen P contents varied from 260 to 4900, and 5 to 740mgkg-1, respectively, while PSol concentrations were between 0.01 and 10.8mgL-1 reflecting the large differences in vegetation history, fertilization schemes, and soil types. Overall, DGT P provided the best correlation with PSol (r2=0.97) demonstrating that DGT P is a versatile measure of P mobility regardless of soil type. Among the DPSs tested, oxalate extractable Al (DPSOx-Al) had the best correlation with PSol (r2=0.87). In the STP versus PSol relationships, STP break-points above which P mobilization increases steeply were 513μgL-1 and 190mgkg-1 for DGT P or Olsen P, respectively, corresponding to PSol concentration of 0.88mgL-1. However, for PSol concentration of 0.1mgL-1 that initiates eutrophication, the corresponding DGT P and Olsen P values were 27μgL-1 and 22mgkg-1, respectively. Over 80% of the investigated soils had DGT P and Olsen P above these values, and thus are at risk of P mobilization threatening receiving waters by eutrophication. This paper demonstrates that the DGT extracted P is a powerful measure for soluble P and hence for assessment of P mobility from a broad range of soil types.

Functional logistic regression is becoming more popular as there are many situations where we are interested in the relation between functional covariates (as input) and a binary response (as output). Several approaches have been advocated, and this paper goes into detail about three of them: dimension reduction via functional principal component analysis, penalized functional regression, and wavelet expansions in combination with Least Absolute Shrinking and Selection Operator penalization. We discuss the performance of the three methods on simulated data and also apply the methods to data regarding lameness detection for horses. Emphasis is on classification performance, but we also discuss estimation of the unknown parameter function.

Lipid flippases of the P4 ATPase family establish phospholipid asymmetry in eukaryotic cell membranes and are involved in many essential cellular processes. The yeast Saccharomyces cerevisiae contains five P4 ATPases, among which Dnf3p is poorly characterized. Here, we demonstrate that Dnf3p is a flippase that catalyzes translocation of major glycerophospholipids, including phosphatidylserine, towards the cytosolic membrane leaflet. Deletion of the genes encoding Dnf3p and the distantly related P4 ATPases Dnf1p and Dnf2p results in yeast mutants with aberrant formation of pseudohyphae, suggesting that the Dnf1p-Dnf3p proteins have partly redundant functions in the control of this specialized form of polarized growth. Furthermore, as previously demonstrated for Dnf1 and Dnf2p, the phospholipid flipping activity of Dnf3p is positively regulated by flippase kinase 1 (Fpk1p) and Fpk2p. Phylogenetic analyses demonstrate that Dnf3p belongs to a subfamily of P4 ATPases specific for fungi and are likely to represent a hallmark of fungal evolution.

Lipid flippases of the P4 ATPase family establish phospholipid asymmetry in eukaryotic cell membranes and are involved in many essential cellular processes. The yeast

Cancer cells exhibit decreased stiffness. We provide novel measurement of cell stiffness changes during transformation. Stiffness decreases as transformation progresses, and RSD distinguishes later stages from earlier stages. Stiffness may be a valuable biomarker for early cancer detection., Cancer cells consistently exhibit decreased stiffness; however, the onset and progression of this change have not been characterized. To study the development of cell stiffness changes, we evaluated the shear stiffness of populations of cells during transformation to a carcinogenic state. Bronchial epithelial cells were exposed to sodium arsenite to initiate early stages of transformation. Exposed cells were cultured in soft agar to further transformation and select for clonal populations exhibiting anchorage-independent growth. Shear stiffness of various cell populations in G1 was assessed using a novel non-invasive assay that applies shear stress with fluid flow and evaluates nanoscale deformation using quantitative phase imaging (QPI). Arsenic-treated cells exhibited reduced stiffness relative to control cells, while arsenic clonal lines, selected by growth in soft agar, were found to have reduced stiffness relative to control clonal lines, which were cultured in soft agar but did not receive arsenic treatment. The relative standard deviation (RSD) of the stiffness of Arsenic clones was reduced compared with control clones, as well as to the arsenic-exposed cell population. Cell stiffness at the population level exhibits potential to be a novel and sensitive framework for identifying the development of cancerous cells.

A classification problem with a functional predictor is studied, and it is suggested to use a multinomial functional regression (MFR) model for the analysis. The discrete wavelet transform and LASSO penalization are combined for estimation, and the fitted model is used for classification of new curves with unknown class membership. The MFR approach is applied to two datasets, one regarding lameness detection for horses and another regarding speech recognition. In the applications, as well as in a simulation study, the performance of the MFR approach is compared to that of other methods for supervised classification of functional data, and MFR performs as well or better than the other methods.

External sourcing from foreign suppliers is an important aspect of the firm’s internationalization However, data on such sourcing is available from neither databases nor annual reports. Thus, the corporate risk implications of such sourcing have not been studied previously. We obtain the necessary data by surveying Scandinavian non-financial firms. We find that highly international firms reduce corporate risk by externally sourcing from foreign suppliers both compared to sourcing from own production facilities abroad (due to superior flexibility) and compared to domestic sourcing (due to off-setting cash flows). Our results are statistically significant, are economically meaningful, and have important policy implications. External sourcing from foreign suppliers is an important aspect of the firm's internationalization. However, data on such sourcing is available from neither databases nor annual reports. Thus, the corporate risk implications of such sourcing have not been studied previously. We obtain the necessary data by surveying Scandinavian non-financial firms. We find that highly international firms reduce corporate risk by externally sourcing from foreign suppliers both compared to sourcing from own production facilities abroad (due to superior flexibility) and compared to domestic sourcing (due to off-setting cash flows). Our results are statistically significant, are economically meaningful, and have important policy implications.

Hybridisation barriers are likely to evolve during allopatric separation of populations, in parallel with divergent adaptation to different conditions in the two ranges. If the populations secondarily come into contact, limited interbreeding between them may affect the subsequent spread of the two lineages and their adaptations in the region of sympatry. Barbarea vulgaris include two genetically divergent lineages that differ in secondary metabolites and resistances to insects and a pathogen. The two plant types grow in different Eurasian ranges but co-occur in Denmark and neighbouring countries, posing the question why they have not merged and the resistances spread from one type into the range of the other. Here, we tested whether a hybridisation barrier contribute to this. Different proportions of plants of the two types were placed in net tents and pollinated by flies, and paternity of the resulting seeds determined with genetic markers. Lower proportions of fruits and seeds developed successfully in mixtures with higher proportions of heterotypic plants (i.e. of the other plant type). When combined with results on offspring paternity in a statistical analysis, we found that heterotypic pollen was much less successful in fertilizing embryos and that heterotypic seeds survived less frequently than the contypic. Mature F1 hybrids in addition produced lower proportions of mature pollen. The two B. vulgaris plant types are thus separated by substantial prezygotic and postzygotic barriers, as strong as reported for crosses between closely related but taxonomically recognised plant species. This may explain why the two B. vulgaris types have not merged in sympatry and why genes for insect-resistance have not introgressed to any extent from one to the other.

Evolution of ecotypic differentiation and maintenance of divergence in sympatry is more likely if populations occupy different habitats. Two genetically divergent chemotypes of Barbarea vulgaris (Brassicaceae) with contrasting insect and disease resistances (G- and P-type) occupy different geographical ranges in Eurasia but co-occur in Denmark. Here, they grow mostly in separate populations, posing the question why they have not merged. In a 2-year reciprocal transplant experiment, we tested whether the two plant types prefer different habitats and are adapted to these, possibly influenced by selection by herbivores. Vegetation and soil analyses indicated that the G-type occurs in slightly drier, less productive and more alkaline sites than the P-type. Plants planted at sites of their own chemotype had a higher biomass in the first year than plants of the non-resident chemotype, but were more damaged by beetle herbivory. This suggests that herbivores have adapted genetically or plastically to the resident chemotype. G-plants were less damaged by molluscs than P-plants, which may be caused by their content of saponins or glucosinolates. Levels of glucosinolates in siliques were higher in G-plants, but varied strongly among sites. Survival, reproduction and biomass in the second year did not differ between plants growing at sites of their own chemotype and plants of the other chemotype. Thus, the two chemotypes of B. vulgaris are adapted to different habitats to some extent, which could contribute to maintain their differentiation in sympatry. Herbivory by some insects, however, may counteract this.

Summary Social interactions in which bacteria respond to one another by modifying their phenotype are central determinants of microbial communities. It is known that interspecific interactions influence the biofilm phenotype of bacteria; a phenotype that is central to the fitness of bacteria. However, the underlying role of fundamental ecological factors, specifically coexistence and phylogenetic history, in biofilm formation remains unclear. This study examines how social interactions affect biofilm formation in multi-species co-cultures from five diverse environments. We found prevalence of increased biofilm formation among co-cultured bacteria that have coexisted in their original environment. Conversely, when randomly co-culturing bacteria across these five consortia, we found less biofilm induction and a prevalence of biofilm reduction. Reduction in biofilm formation was even more predominant when co-culturing bacteria from environments where long-term coexistence was unlikely to have occurred. Phylogenetic diversity was not found to be a strong underlying factor but a relation between biofilm induction and phylogenetic history was found. The data indicates that biofilm reduction is typically correlated with an increase in planktonic cell numbers, thus implying a behavioral response rather than mere growth competition. Our findings suggest that an increase in biofilm formation is a common adaptive response to long-term coexistence.

Edible insects have been promoted as an alternative sustainable source of protein, however many are still unwilling to eat insects due to the perceived disgust and neophobia. In light of the growing demand for alternative protein sources, this study examined edible insects. Tactile stimuli with food have been demonstrated to be an alternative strategy to reduce children’s natural neophobic reactions. The present study aims to examine the potential of tactile interactions in the form of a cooking activity to introduce edible insects to children. To this purpose, two types of insects (grasshopper and mealworm) were incorporated into a traditional Danish snack (oatmeal balls). Children (n = 148) tasted and evaluated the food that was either self-prepared or other-prepared. Tasting order and type of insect had significant effects on the hedonic response. The insect oatmeal balls were rated higher in the first tasting compared to the second. The mealworm version of the oatmeal balls received higher hedonic ratings than the grasshopper version. This we propose, is caused by different degrees of ‘animalness’ of the two insects. The cooking activity did not have a significant effect on children's hedonic response to the insect oatmeal balls.

Cropping practice influence arthropod species abundance and diversity in agroecosystems, thus impacting populations of insect pests and their natural enemies. A field study was conducted to investigate whether pest and natural enemy abundance was higher in organic compared with conventional strawberry fields, and whether the non-managed vegetation surrounding the field impacted natural enemy abundance and diversity in and around the field sites. The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), and predatory mites were sampled from strawberry leaflets and the leaves of wild herbaceous plants from the surrounding vegetation. Insect predators, hymenopteran parasitoids, spiders and herbivores were collected from the same habitats. Abundance of T. urticae was ten-fold higher in conventional compared to organic strawberry fields, whereas the ratio of spider mite to predatory mites was 9.5 times lower in organic compared with conventional strawberry fields. The 14 species of predatory mites identified were all from the family Phytoseiidae, and predominantly found on stinging nettle, Urtica dioica L. (Rosales: Urticaceae) (> 80% of individuals), in the surrounding vegetation. There was no significant effect of cropping practice on the density of insect predators and spiders, while the density of insect parasitoids and insect herbivores was higher in the organic compared to the conventional sites. Total species richness at the edge of organic fields and in the surrounding vegetation of both organic and conventional sites was higher than in the other sampling sites. This study demonstrates the major impact of cropping practice on T. urticae abundance and on the prey to predator ratio, whilst also emphasizing the importance of the surrounding vegetation as a source of natural enemies of phytophagous arthropods.

Application of wood ash to forest soils is used to counteract acidification and to reintroduce nutrients otherwise lost by harvesting, particularly in silvicultural systems with intensified biomass removal. pH governs many essential processes like nutrient availability for plant uptake, heavy metal mobility and ecosystem functioning in forest soil, but changes in soil pH on a micro scale after ash application have not been reported. The aim of this study was therefore to investigate micro vertical changes in soil pH and base cations over time after application of wood ash on forest soil. A column experiment with repacked O- and A-horizon forest soil was constructed and treatments of 0, 3 and 9 t wood ash ha−1 were applied on top of the soil in replicates of three. Soil cores were extracted from the columns monthly over one year for pH micro sensor measurements of O- and A-horizon (0–5 cm soil depth) and at four occasions (62, 152, 278 and 363 days after treatment) additional soil cores were extracted for base cation analysis. Results showed that ash application creates strong pH gradients ranging from pH 11 to 4.5 within the top 1–1.5 cm of the soil, with the highest variation within the first 50 days after application. Application of ash increases the nutrient availability. If micro scale variation is not considered, mean pH values can be a misleading indicator for the extreme conditions that can exists in the top part of the soil after ash application.

Wetland soils may act as sinks for phosphorus from agricultural soils but P sorption is sensitive to soil redox conditions. When soils become flooded and anoxic, FeIII is reduced and P sorption decreases. To get improved knowledge about the relationship between redox state and P solubility under field conditions, a grid of 4 × 5 sampling points laid out in an Fe-rich meadow soil bordering a Danish river were monitored during 11 weeks in the spring comprising in total five sampling dates. The redox state was quantified as the degree of Fe reduction (DRFe), i.e. percentage of soluble plus sorbed FeII to total pedogenic, non-silicate Fe. Average DRFe was highest (76 %) at beginning of April and decreased to 36 % within 9 weeks. The temporal and spatial variation of DRFe and the soil solution concentrations of Fe and P were high. Thus, average P in solution (Psol) ranged from 24 to 4 μM, closely correlated with average DRFe (R 2 = 0.83). Soils farthest away from the river drained first and hence exhibited the biggest change in DRFe and Psol over time. The study demonstrates the great spatial and temporal variability of redox state and P solubility in such wetlands. Some likely consequences of redox oscillations are indicated.

Ash dieback disease (caused by Hymenoscyphus fraxineus) has affected European ash species (Fraxinus spp.) in recent decades. However, some Asian and American species of Fraxinus and certain genotypes of Fraxinus excelsior are less affected by the disease. We used ITS1-metabacoding to explore the drivers influencing diversity and composition of the twig fungal communities of Fraxinus species and F. excelsior genotypes. Our results revealed that fungi in the classes Eurotiomycetes and Dothideomycetes were among the most prevalent taxa in both Fraxinus species and F. excelsior genotypes. The diversity of the fungal communities differed significantly among Fraxinus species and could be explained by seed origin. Neither host genotype nor season had a significant effect on the community diversity of F. excelsior genotypes. On the other hand, the composition of twig fungal communities differed significantly among host species and among F. excelsior genotypes, and in F. excelsior there was also a significant effect of season on the composition of the fungal community. We did not find a clear effect of ash dieback susceptibility on either diversity or composition of fungal communities in twigs of Fraxinus species, although the effect was significant on the composition of fungal communities among F. excelsior genotypes. Our results demonstrated differences in fungal communities among species of Fraxinus and of F. excelsior genotypes, suggesting specific relationship between individual host genotypes and endophytic fungi.

Cancer cells consistently exhibit decreased stiffness; however, the onset and progression of this change have not beencharacterized. To study the development of cell stiffness changes, we evaluated the shear stiffness of populations ofcells during transformation to a carcinogenic state. Bronchial epithelial cells were exposed to sodium arsenite to initiateearly stages of transformation. Exposed cells were cultured in soft agar to further transformation and select for clonalpopulations exhibiting anchorage-independent growth. Shear stiffness of various cell populations in G1 was assessed usinga novel non-invasive assay that applies shear stress with fluid flow and evaluates nanoscale deformation using quantitativephase imaging (QPI). Arsenic-treated cells exhibited reduced stiffness relative to control cells, while arsenic clonal lines,selected by growth in soft agar, were found to have reduced stiffness relative to control clonal lines, which were cultured insoft agar but did not receive arsenic treatment. The relative standard deviation (RSD) of the stiffness of Arsenic clones wasreduced compared with control clones, as well as to the arsenic-exposed cell population. Cell stiffness at the populationlevel exhibits potential to be a novel and sensitive framework for identifying the development of cancerous cells.

This paper presents a study on symmetry of repeated bi-phased data signals, in particular, on quantification of the deviation between the two parts of the signal. Three symmetry scores are defined using functional data techniques such as smoothing and registration. One score is related to the L 2-distance between the two parts of the signal, whereas the other two are constructed to specifically measure differences in amplitude and phase. Moreover, symmetry scores based on functional principal component analysis (PCA) are examined. The scores are applied to acceleration signals from a study on equine gait. The scores turn out to be highly associated with lameness, and their applicability for lameness quantification and detection is investigated. Four classification approaches turn out to give similar results. The scores describing amplitude and phase variation turn out to outperform the PCA scores when it comes to the classification of lameness.

We suggest a simple isobole analysis for binary mixture toxicity experiments. The analysis is based on estimated logarithmic effect concentrations and their corresponding standard errors. The suggested model allows for synergism/antagonism and incorporates within-mixture variation as well as between-mixture variation in a random effects model. The likelihood ratio test for the hypothesis of concentration addition (CA) is examined, in particular its small sample properties. We study two datasets on the joint effect of acifluorfen versus diquat and glyphosate versus mechlorprop, respectively, on the growth of the aquatic macrophyte Lemna minor.

Summary Reasons for performing study: Equine lameness examination is based on subjective visual scoring of lameness. Instrumented objective methods for lameness examinations may be complicated to perform and the equipment is often stationary. Accelerometry has a potential clinical use; however, the reduction and interpretation of equine accelerometric data are not yet routine and the value of accelerometry in equine lameness examination is unclear. Objectives: To use accelerometric data to calculate 2 different accelerometric symmetry scores and to evaluate the agreement of these with traditional lameness scores done by experienced equine practitioners. Methods: Six sound horses were equipped with a 3 axis 10G piezoresistant accelerometer at the lowest point of the back. Horses were trotted and video recorded at 0, 3, 15, 30, 45 and 60 min after injection of saline into one metacarpophalangeal joint. Video recordings were scored in a blind manner according to the AAEP scale by 2 experienced practitioners. Interobserver agreements and 2 symmetry scores S and A, developed on the basis of Fourier transformation of the obtained accelerometric data, were calculated and regression analysis between AAEP scores and symmetry scores was performed. Results: Interobserver agreements were 70%. There was a statistically significant relationship between AAEP lameness scores and both symmetry scores. Conclusions: Both symmetry scores showed a significant relationship with the AAEP scores and can be a valuable tool in the detection and quantification of lameness. While the S score was able to detect changes in degree of lameness, the A score was capable of detecting the lame diagonal. However, more research is needed for the development of a combined accelerometric score to take advantage of the strengths of each of the symmetry scores.

A goodness-of-fit test for continuous-time models is developed that examines if the parameter estimates are consistent with another for different sampling frequencies. The test compares parameter estimates obtained from estimating functions for downsamples of the data. We prove asymptotic results for stationary and ergodic processes, and apply the downsampling test to linear drift diffusions. Simulations indicate that the test is quite powerful in detecting non-Markovian deviations from the linear drift diffusions.

Simultaneous equations have become increasingly popular for describing the effects of nutri- tion on the utilization of ME for protein (PD) and lipid deposition (LD) in animals. The study developed a multivariate nonlinear mixed effects (MNLME) frame- work and compared it with an alternative method for estimating parameters in simultaneous equations that described energy metabolism in growing pigs, and then proposed new PD and LD equations. The general sta- tistical framework was implemented in the NLMIXED procedure in SAS. Alternative PD and LD equations were also developed, which assumed that the instan- taneous response curve of an animal to varying energy supply followed the law of diminishing returns behavior. The Michaelis-Menten function was adopted to repre- sent a biological relationship in which the affinity con- stant (k) represented the sensitivity of PD to ME above maintenance. The approach accommodated inclusion of a PD potential (PDPotential) concept. This was described by a Gompertz function, which was parameterized in terms of the maximum rate of PD (PDMax) and corre- sponding BW (BWPDMax) at that point. Metabolizable energy for LD was equated to the difference between ME intake and the sum of ME used for maintenance and PD. Metabolizable energy designated for PD and LD was used, with efficiencies kp and kf, respectively. The new equations were compared with the van Mil- gen and Noblet (1999) equations using 2 comprehensive data sets on energy metabolism in growing pigs. The 2 equation sets were evaluated using information cri- teria, which showed that the new equations performed best for data set II, whereas the reverse was true for the first. For the data set I population, estimates for kp and kf were 0.57 (SE = 0.05) and 0.84 (SE = 0.03), respectively. Maintenance was quantified as 1.10 (SE = 0.08) MJ/d· kg 0.55 . The animal variation in the param- eter kp was estimated to be 6% CV. The animal varia- tion in PDMax and kf was estimated to be 9 and 10% of the population estimates, respectively. It was con- cluded that application of the MNLME framework was superior to the multivariate nonlinear regression model because the MNLME method accounted for correlated errors associated with PD and LD measurements and could also include the random effect of animal. It is rec- ommended that multivariate models used to quantify energy metabolism in growing pigs should account for animal variability and correlated measurement errors.