Plenary discussion on revision of the EPPO guidelines/risk assessment scheme

contribution to session I Regulatory issue

Review of honeybee pesticide poisoning incidents in Europe – evaluation of the hazard quotient approach for risk assessment

contribution to session V Honey bee poisoning incidents and monitoring schemes Background: Honeybee risk assessment is required in Europe for all pesticides where bees may be exposed. This is well established for sprayed products where the hazard quotient (HQ), calculated by dividing the application rate of the sprayed product active ingredient by the LD50, is less than 50 the product is considered safe to bees (unless it is an IGR). In the UK, Germany and the Netherlands post-registration monitoring schemes on the poisoning of honeybee by pesticides collate data on honeybee incidents. Results: The incident schemes have been invaluable in identifying agronomic practices resulting in honeybee mortality and changes have been made to labelling to address such issues. The decrease in the numbers of incidents reported supports the assertion that such schemes have positively contributed to the regulatory process and also provide confidence in the risk assessment approaches. Conclusion: This review of incidents in Europe over the last 25 years suggests that the HQ approach to risk assessment for honeybees offers an appropriate level of protection. Keywords: honeybee, pesticide, hazard quotient, risk assessmen

Preparando estudantes da Pós-Graduação para a indústria e aprendizado para a vida: uma abordagem baseada em projeto

Este estudo de caso baseia-se em projetos que têm uma dimensão de mundo real, em que os alunos estão mais propensos a adquirir as competências e habilidades que a indústria exige. Informações de base são apresenta- das, relacionando vínculos entre a academia, a indústria e o papel dos projetos dos estudantes. Os dados são provenien- tes de programas de computação em nível de mestrado, da Universidade de Sunderland, e dos projetos desenvolvidos por cada um dos estudantes. Uma abordagem específica permitiu que centenas de projetos bem sucedidos, realiza- dos anualmente com o setor da indústria, fossem descritos juntamente com a avaliação de sua eficácia em dar aos alunos uma formação de competência para a vida

A new mixture copula model for spatially correlated multiple variables with an environmental application

In environmental monitoring, multiple spatial variables are often sampled at a geographical location that can depend on each other in complex ways, such as non-linear and non-Gaussian spatial dependence. We propose a new mixture copula model that can capture those complex relationships of spatially correlated multiple variables and predict univariate variables while considering the multivariate spatial relationship. The proposed method is demonstrated using an environmental application and compared with three existing methods. Firstly, improvement in the prediction of individual variables by utilising multivariate spatial copula compares to the existing univariate pair copula method. Secondly, performance in prediction by utilising mixture copula in the multivariate spatial copula framework compares with an existing multivariate spatial copula model that uses a non-linear principal component analysis. Lastly, improvement in the prediction of individual variables by utilising the non-linear non-Gaussian multivariate spatial copula model compares to the linear Gaussian multivariate cokriging model. The results show that the proposed spatial mixture copula model outperforms the existing methods in the cross-validation of actual and predicted values at the sampled locations

Differential Hox expression in murine embryonic stem cell models of normal and malignant hematopoiesis

The Hox family are master transcriptional regulators of developmental processes, including hematopoiesis. The Hox regulators, caudal homeobox factors (Cdx1-4), and Meis1, along with several individual Hox proteins, are implicated in stem cell expansion during embryonic development, with gene dosage playing a significant role in the overall function of the integrated Hox network. To investigate the role of this network in normal and aberrant, early hematopoiesis, we employed an in vitro embryonic stem cell differentiation system, which recapitulates mouse developmental hematopoiesis. Expression profiles of Hox, Pbx1, and Meis1 genes were quantified at distinct stages during the hematopoietic differentiation process and compared with the effects of expressing the leukemic oncogene Tel/PDGFR;2. During normal differentiation the Hoxa cluster, Pbx1 and Meis1 predominated, with a marked reduction in the majority of Hox genes (27/39) and Meis1 occurring during hematopoietic commitment. Only the posterior Hoxa cluster genes (a9, a10, a11, and a13) maintained or increased expression at the hematopoietic colony stage. Cdx4, Meis1, and a subset of Hox genes, including a7 and a9, were differentially expressed after short-term oncogenic (Tel/PDGFR;2) induction. Whereas Hoxa4-10, b1, b2, b4, and b9 were upregulated during oncogenic driven myelomonocytic differentiation. Heterodimers between Hoxa7/Hoxa9, Meis1, and Pbx have previously been implicated in regulating target genes involved in hematopoietic stem cell (HSC) expansion and leukemic progression. These results provide direct evidence that transcriptional flux through the Hox network occurs at very early stages during hematopoietic differentiation and validates embryonic stem cell models for gaining insights into the genetic regulation of normal and malignant hematopoiesis

Lower pollen nutritional quality delays nest building and egg laying in Bombus terrestris audax micro-colonies leading to reduced biomass gain

This version of the article has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at: http://dx.doi.org/10.1007/s13592-021-00885-3The performance of Bombus terrestris micro-colonies fed five diets differing in pollen species composition and level of nine essential amino acids (EAA; leucine, lysine, valine, arginine, isoleucine, phenylalanine, threonine, histidine, methionine) was assessed for 37 days by recording total biomass gain, nest building initiation, brood production (eggs, small and large larvae, pupae, drones), nectar, and pollen collection. Stronger colony performance was linked to higher amino acid levels but no consistent differences in biomass gain were recorded between mono- and poly-species diets. Poorest performance occurred in micro-colonies offered pure oilseed rape (OSR) pollen which contained the lowest EAA levels. Reduced micro-colony development (delayed nest initiation and lower brood production) was related to OSR proportion in the diet and lower EAA levels. Results are discussed in relation to selection of plant species in the design of habitats to promote bee populations

Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites

<p>Abstract</p> <p>Background</p> <p>Sensitive detection of parasite surface antigens expressed on erythrocyte membranes is necessary to further analyse the molecular pathology of malaria. This study describes a modified biotin labelling/osmotic lysis method which rapidly produces membrane extracts enriched for labelled surface antigens and also improves the efficiency of antigen recovery compared with traditional detergent extraction and surface radio-iodination. The method can also be used with <it>ex-vivo </it>parasites.</p> <p>Methods</p> <p>After surface labelling with biotin in the presence of the inhibitor furosemide, detergent extraction and osmotic lysis methods of enriching for the membrane fractions were compared to determine the efficiency of purification and recovery. Biotin-labelled proteins were identified on silver-stained SDS-polyacrylamide gels.</p> <p>Results</p> <p>Detergent extraction and osmotic lysis were compared for their capacity to purify biotin-labelled <it>Plasmodium falciparum </it>and <it>Plasmodium chabaudi </it>erythrocyte surface antigens. The pellet fraction formed after osmotic lysis of <it>P. falciparum-infected </it>erythrocytes is notably enriched in suface antigens, including PfEMP1, when compared to detergent extraction. There is also reduced co-extraction of host proteins such as spectrin and Band 3.</p> <p>Conclusion</p> <p>Biotinylation and osmotic lysis provides an improved method to label and purify parasitised erythrocyte surface antigen extracts from both <it>in vitro </it>and <it>ex vivo Plasmodium </it>parasite preparations.</p

Mining Milk for Factors which Increase the Adherence of Bifidobacterium longum subsp. infantis to Intestinal Cells

peer-reviewedBifidobacteria play a vital role in human nutrition and health by shaping and maintaining the gut ecosystem. In order to exert a beneficial effect, a sufficient population of bifidobacteria must colonise the host. In this study, we developed a miniaturised high-throughput in vitro assay for assessing the colonising ability of bacterial strains in human cells. We also investigated a variety of components isolated from different milk sources for their ability to increase the adherence of Bifidobacterium longum subsp. infantis ATCC 15697, a common member of the gastrointestinal microbiota of breastfed infants, to HT-29 cells. Both conventional and miniaturised colonisation assays were employed to examine the effect of 13 different milk-derived powders on bacterial adherence, including positive controls which had previously resulted in increased bifidobacterial adherence (human milk oligosaccharides and a combination of 3′- and 6′-sialylactose) to intestinal cells. Immunoglobulin G enriched from bovine whey and goat milk oligosaccharides resulted in increased adhesion (3.3- and 8.3-fold, respectively) of B. infantis to the intestinal cells and the miniaturised and conventional assays were found to yield comparable and reproducible results. This study highlights the potential of certain milk components to favourably modulate adhesion of bifidobacteria to human intestinal cells

A review of the factors that influence pesticide residues in pollen and nectar: future research requirements for optimising the estimation of pollinator exposure

In recent years, the impact of Plant Protection Products (PPPs) on insect pollinator decline has stimulated significant amounts of research, as well as political and public interest. PPP residues have been found in various bee-related matrices, resulting in governmental bodies worldwide releasing guidance documents on methods for the assessment of the overall risk of PPPs to different bee species. An essential part of these risk assessments are PPP residues found in pollen and nectar, as they represent a key route of exposure. However, PPP residue values in these matrices exhibit large variations and are not available for many PPPs and crop species combinations, which results in inaccurate estimations and uncertainties in risk evaluation. Additionally, residue studies on pollen and nectar are expensive and practically challenging. An extrapolation between different cropping scenarios and PPPs is not yet justified, as the behaviour of PPPs in pollen and nectar is poorly understood. Therefore, this review aims to contribute to a better knowledge and understanding of the fate of PPP residues in pollen and nectar and to outline knowledge gaps and future research needs. The literature suggests that four primary factors, the crop type, the application method, the physicochemical properties of a compound and the environmental conditions have the greatest influence on PPP residues in pollen and nectar. However, these factors consist of many sub-factors and initial effects may be disguised by different sampling methodologies, impeding their exact characterisation. Moreover, knowledge about these factors is ambiguous and restricted to a few compounds and plant species. We propose that future research should concentrate on identifying relationships and common features amongst various PPP applications and crops, as well as an overall quantification of the described parameters; in order to enable a reliable estimation of PPP residues in pollen, nectar and other bee matrices

Bitopic binding mode of an M1 muscarinic acetylcholine receptor agonist associated with adverse clinical trial outcomes

The realisation of the therapeutic potential of targeting the M1 muscarinic acetylcholine receptor (M1 mAChR) for the treatment of cognitive decline in Alzheimer's disease has prompted the discovery of M1 mAChR ligands showing efficacy in alleviating cognitive dysfunction in both rodents and humans. Among these is GSK1034702, described previously as a potent M1 receptor allosteric agonist, which showed pro-cognitive effects in rodents and improved immediate memory in a clinical nicotine withdrawal test but induced significant side-effects. Here we provide evidence using ligand binding, chemical biology and functional assays to establish that rather than the allosteric mechanism claimed, GSK1034702 interacts in a bitopic manner at the M1 mAChR such that it can concomitantly span both the orthosteric and an allosteric binding site. The bitopic nature of GSK1034702 together with the intrinsic agonist activity and a lack of muscarinic receptor subtype selectivity reported here, all likely contribute to the adverse effects of this molecule in clinical trials. We conclude that these properties, whilst imparting beneficial effects on learning and memory, are undesirable in a clinical candidate due to the likelihood of adverse side effects. Rather, our data supports the notion that "pure" positive allosteric modulators showing selectivity for the M1 mAChR with low levels of intrinsic activity would be preferable to provide clinical efficacy with low adverse responses