Clotting is critical in limiting loss of hemolymph and initiating wound healing in insects as well as in vertebrates. Clotting is also an important immune defense, quickly forming a secondary barrier to infection, thereby immobilizing, and possibly killing bacteria directly. Here, we describe methods to assess clotting and to extract the clot from Drosophila larval hemolymph by using aggregation of paramagnetic beads. The validity of the assay was demonstrated by characterization of mutants. We show that clotting occurs in the absence of phenoloxidase and that the Drosophila clot binds bacteria. We also describe a pullout assay to purify the clot as a whole, free from entrapped hemocytes and cellular debris. Proteins subsequently identified by mass spectrometry include both predicted and novel clot proteins. Immune induction has been shown for three of the latter, namely Tiggrin and two unknown proteins (GC15825 and CG15293) that we now propose function in hemolymph clotting. The most abundant clot protein is Hemolectin, and we confirm that hemolectin mutant larvae show clotting defects.

The changing world of scholarly communication and the emerging new wave of ‘Open Science’ or ‘Open Research’ has brought to light a number of controversial and hotly debated topics. Evidence-based rational debate is regularly drowned out by misinformed or exaggerated rhetoric, which does not benefit the evolving system of scholarly communication. This article aims to provide a baseline evidence framework for ten of the most contested topics, in order to help frame and move forward discussions, practices, and policies. We address issues around preprints and scooping, the practice of copyright transfer, the function of peer review, predatory publishers, and the legitimacy of ‘global’ databases. These arguments and data will be a powerful tool against misinformation across wider academic research, policy and practice, and will inform changes within the rapidly evolving scholarly publishing system.

Germ cells are a population of cells that do not differentiate to form somatic tissue but form the egg and sperm that ensure the reproduction of the organism. To understand how germ cells form, holds a key for identifying what sets them apart from all other cells of the organism. There are large differences between embryos regarding where and when germ cells form but the expression of Vasa protein is a common trait of germ cells. We studied the role of vasa during germ cell formation in the crustacean Parhyale hawaiensis. In a striking difference to the posterior specification of the group of germ cells in the arthropod model Drosophila, all germ cells in Parhyale originate from a single germ line progenitor cell of the 8-cell stage. We found vasa RNA ubiquitously distributed from 1-cell to 16-cell stage in Parhyale and localized to the germ cells from 32-cell stage onwards. Localization of vasa RNA to the germ cells is controlled by its 3'UTR and this could be mimicked by fluorescently labeled 3'UTR RNA. Vasa protein was first detectable at the 100-cell stage. MO-mediated inhibition of vasa translation caused germ cells to die after gastrulation. This means that in Parhyale Vasa protein is not required for the initial generation of the clone of germ cells but is required for their subsequent proliferation and maintenance. It also means that the role of vasa changed substantially during an evolutionary switch in the crustaceans by Parhyale from the specification of a group of germ cells to that of a single germ line progenitor. This is the first functional study of vasa in an arthropod beyond Drosophila.

There is no clear-cut boundary between Free and Open Source Software and Open Scholarship, and the histories, practices, and fundamental principles between the two remain complex. In this study, we critically appraise the intersections and differences between the two movements. Based on our thematic comparison here, we conclude several key things. First, there is substantial scope for new communities of practice to form within scholarly communities that place sharing and collaboration/open participation at their focus. Second, Both the principles and practices of FOSS can be more deeply ingrained within scholarship, asserting a balance between pragmatism and social ideology. Third, at the present, Open Scholarship risks being subverted and compromised by commercial players. Fourth, the shift and acceleration towards a system of Open Scholarship will be greatly enhanced by a concurrent shift in recognising a broader range of practices and outputs beyond traditional peer review and research articles. In order to achieve this, we propose the formulation of a new type of institutional mandate. We believe that there is substantial need for research funders to invest in sustainable open scholarly infrastructure, and the communities that support them, to avoid the capture and enclosure of key research services that would prevent optimal researcher behaviours. Such a shift could ultimately lead to a healthier scientific culture, and a system where competition is replaced by collaboration, resources (including time and people) are shared and acknowledged more efficiently, and the research becomes inherently more rigorous, verified, and reproducible.

The arthropod cuticle is a multilayered extracellular matrix produced by the epidermis during embryogenesis and moulting. Molecularly and histologically, cuticle differentiation has been extensively investigated in the embryo of the insect Drosophila melanogaster. To learn about the evolution of cuticle differentiation, we have studied the histology of cuticle differentiation during embryogenesis of the amphipod crustacean Parhyale hawaiensis, which had a common ancestor with Drosophila about 510 million years ago. The establishment of the layers of the Parhyale juvenile cuticle is largely governed by mechanisms observed in Drosophila, e.g. as in Drosophila, the synthesis and arrangement of chitin in the inner procuticle are separate processes. A major difference between the cuticle of Parhyale and Drosophila concerns the restructuring of the Parhyale dorsal epicuticle after deposition. In contrast to the uniform cuticle of the Drosophila larva, the Parhyale cuticle is subdivided into two regions, the ventral and the dorsal cuticles. Remarkably, the boundary between the ventral and dorsal cuticles is sharp suggesting active extracellular regionalisation. The present analysis of Parhyale cuticle differentiation should allow the characterisation of the cuticle-producing and -organising factors of Parhyale (by comparison with the branchiopod crustacean Daphnia pulex) in order to contribute to the elucidation of fundamental questions relevant to extracellular matrix organisation and differentiation.