Nuclear Factors Bind to a Conserved DNA Element That Modulates Transcription of Anopheles gambiae Trypsin Genes

The Anopheles gambiae trypsin family consists of seven genes that are transcribed in the gut of female mosquitoes in a temporal coordinated and mutually exclusive manner, suggesting the involvement of a complex transcription regulatory mechanism. We identified a highly conserved 12-nucleotide motif present in all A. gambiae and Anopheles stephensi trypsin promoters. We investigated the role of this putative trypsin regulatory element (PTRE) in controlling the transcription of the trypsin genes. Gel shift experiments demonstrated that nuclear proteins of A. gambiae cell lines formed two distinct complexes with probes encompassing the PTRE sequence. Mapping of the binding sites revealed that one of the complex has the specificity of a GATA transcription factor. Promoter constructs containing mutations in the PTRE sequence that selectively abolished the binding of either one or both complexes exerted opposite effects on the transcriptional activity of trypsin promoters in A. gambiae and Aedes aegypti cell lines. In addition, the expression of a novel GATA gene was highly enriched in A. gambiae guts. Taken together our data prove that factors binding to the PTRE region are key regulatory elements possibly involved in the blood meal-induced repression and activation of transcription in early and late trypsin genes

Photo 3D technology applied to e-Learning tools production for animal biology

[EN] The teachers from the Biology Department of the University of Lille have been developing for the last two years, a digitalization program of naturalist collections. This project aims to preserve this scientific heritage and to use it for animal and plant biology teaching. Specimens are digitalized by a photo 3D capture system, that produces a 360° and/or hemispheric images of the objects starting from high-resolution pictures. Based on the use of this particular imaging technology, teachers realized multimedia eBooks and a series of files “at the glance“ for practical works in animal biology for Bachelor students in Life Sciences. These supports, enriched by graphic complements, texts, legends and interactive animations, are available on the pedagogic platform Moodle. These digital tools are viewable on computers and can be handled on smartphones and tablets for a nomad utilisation. Students generally consider these supports useful for learning and they consult these resources before, during and after the practical sessions. This distance-learning approach gives the students a complete autonomy for practical session preparation and reviews. The innovative tools here presented constitute a useful learning complement to classical academic lectures in animal biology.The project “Mammal teeth and diets“ is funded by UNISCIEL (Université des Sciences en ligne, http://www.unisciel.fr).Sautière, P.; Delbende, C.; Deleplanque, B.; Mikolajczyk, B.; Vizioli, J. (2019). Photo 3D technology applied to e-Learning tools production for animal biology. En HEAD'19. 5th International Conference on Higher Education Advances. Editorial Universitat Politècnica de València. 921-928. https://doi.org/10.4995/HEAD19.2019.9225OCS92192

Interaction of HmC1q with leech microglial cells: involvement of C1qBP-related molecule in the induction of cell chemotaxis

<p>Abstract</p> <p>Background</p> <p>In invertebrates, the medicinal leech is considered to be an interesting and appropriate model to study neuroimmune mechanisms. Indeed, this non-vertebrate animal can restore normal function of its central nervous system (CNS) after injury. Microglia accumulation at the damage site has been shown to be required for axon sprouting and for efficient regeneration. We characterized <it>Hm</it>C1q as a novel chemotactic factor for leech microglial cell recruitment. In mammals, a C1q-binding protein (C1qBP alias gC1qR), which interacts with the globular head of C1q, has been reported to participate in C1q-mediated chemotaxis of blood immune cells. In this study, we evaluated the chemotactic activities of a recombinant form of <it>Hm</it>C1q and its interaction with a newly characterized leech C1qBP that acts as its potential ligand.</p> <p>Methods</p> <p>Recombinant <it>Hm</it>C1q (r<it>Hm</it>C1q) was produced in the yeast <it>Pichia pastoris</it>. Chemotaxis assays were performed to investigate r<it>Hm</it>C1q-dependent microglia migration. The involvement of a C1qBP-related molecule in this chemotaxis mechanism was assessed by flow cytometry and with affinity purification experiments. The cellular localization of C1qBP mRNA and protein in leech was investigated using immunohistochemistry and <it>in situ </it>hybridization techniques.</p> <p>Results</p> <p>r<it>Hm</it>C1q-stimulated microglia migrate in a dose-dependent manner. This r<it>Hm</it>C1q-induced chemotaxis was reduced when cells were preincubated with either anti-<it>Hm</it>C1q or anti-human C1qBP antibodies. A C1qBP-related molecule was characterized in leech microglia.</p> <p>Conclusions</p> <p>A previous study showed that recruitment of microglia is observed after <it>Hm</it>C1q release at the cut end of axons. Here, we demonstrate that r<it>Hm</it>C1q-dependent chemotaxis might be driven via a <it>Hm</it>C1q-binding protein located on the microglial cell surface. Taken together, these results highlight the importance of the interaction between C1q and C1qBP in microglial activation leading to nerve repair in the medicinal leech.</p

Cross-Species Single-Cell Analysis Reveals Divergence of the Primate Microglia Program

Summary Microglia, the brain-resident immune cells, are critically involved in many physiological and pathological brain processes, including neurodegeneration. Here we characterize microglia morphology and transcriptional programs across ten species spanning more than 450 million years of evolution. We find that microglia express a conserved core gene program of orthologous genes from rodents to humans, including ligands and receptors associated with interactions between glia and neurons. In most species, microglia show a single dominant transcriptional state, whereas human microglia display significant heterogeneity. In addition, we observed notable differences in several gene modules of rodents compared with primate microglia, including complement, phagocytic, and susceptibility genes to neurodegeneration, such as Alzheimer’s and Parkinson’s disease. Our study provides an essential resource of conserved and divergent microglia pathways across evolution, with important implications for future development of microglia-based therapies in humans

Allograft Inflammatory Factor-1 in Metazoans: Focus on Invertebrates

Allograft inflammatory factor-1 (AIF-1) is a calcium-binding scaffold/adaptor protein often associated with inflammatory diseases. Originally cloned from active macrophages in humans and rats, this gene has also been identified in other vertebrates and in several invertebrate species. Among metazoans, AIF-1 protein sequences remain relatively highly conserved. Generally, the highest expression levels of AIF-1 are observed in immunocytes, suggesting that it plays a key role in immunity. In mammals, the expression of AIF-1 has been reported in different cell types such as activated macrophages, microglial cells, and dendritic cells. Its main immunomodulatory role during the inflammatory response has been highlighted. Among invertebrates, AIF-1 is involved in innate immunity, being in many cases upregulated in response to biotic and physical challenges. AIF-1 transcripts result ubiquitously expressed in all examined tissues from invertebrates, suggesting its participation in a variety of biological processes, but its role remains largely unknown. This review aims to present current knowledge on the role and modulation of AIF-1 and to highlight its function along the evolutionary scale

Allograft inflammatory factor-1 in metazoans: focus on invertebrates

International audienceAllograft inflammatory factor-1 (AIF-1) is a calcium-binding scaffold/adaptor protein often associated with inflammatory diseases. Originally cloned from active macrophages in humans and rats, this gene has also been identified in other vertebrates and in several invertebrate species. Among metazoans, AIF-1 protein sequences remain relatively highly conserved. Generally, the highest expression levels of AIF-1 are observed in immunocytes, suggesting that it plays a key role in immunity. In mammals, the expression of AIF-1 has been reported in different cell types such as activated macrophages, microglial cells, and dendritic cells. Its main immunomodulatory role during the inflammatory response has been highlighted. Among invertebrates, AIF-1 is involved in innate immunity, being in many cases upregulated in response to biotic and physical challenges. AIF-1 transcripts result ubiquitously expressed in all examined tissues from invertebrates, suggesting its participation in a variety of biological processes, but its role remains largely unknown. This review aims to present current knowledge on the role and modulation of AIF-1 and to highlight its function along the evolutionary scale

Laboratório HE.SU.TECH.: pesquisa acadêmica combinada com a prática aplicada na documentação, representação e preservação do patrimônio arquitetônico - o caso da Villa Forni Cerato de Paladdio

Este artigo traz um estudo aprofundado sobre documentação, representação e práxis de conservação do patrimônio arquitetônico no campo da pesquisa acadêmica aplicada. A ênfase são os avanços tecnológicos na documentação digital aplicada ao Patrimônio Cultural. Destaca-se a importância em se conciliar o conhecimento da pesquisa acadêmica com o trabalho técnico aplicado. O mapeamento arquitetônico é uma das primeiras ações realizadas no processo de conhecimento de um bem e, como tal, ele torna-se imprescindível para as subsequentes operações de conservação, restauração e gestão. Este artigo apresenta uma experiência de técnicas integradas de mapeamento digital para a digitalização de uma arquitetura de Andrea Palladio: a Villa Forni Cerato. Esta experiência foi realizada pelo Grupo de Pesquisa Heritage Survey Technology (HE.SU.TECH.) do Politecnico di Milano - Polo di Mantova que desenvolve atividades de pesquisa e consultoria com instituições ligadas ao patrimônio italiano desde 2000. O caso da Villa Forni Cerato é particularmente interessante porque, neste caso, a digitalização foi integrada com outras fontes documentais, tais como as plantas e secções originais da própria Villa. O levantamento da Villa exigiu a integração de muitas tecnologias diferentes usadas no campo geomático para o Patrimônio Cultural. Este estudo de caso, em particular, tornou-se uma oportunidade para testar técnicas fotogramétricas com diferentes conjuntos fotográficos, adquiridos tanto do solo como por meio de Sistemas Aéreos Não Tripulados (UAS). Este trabalho tem sido objeto de discussão dentro do projeto de intercâmbio entre as duas universidades: Politécnico de Milão e Universidade de São Paulo.This article runs an in-depth examination of documentation, representation and conservation praxis of Architectural Heritage in the field of applied academic research. The focus is put on technological advances in digital documentation applied to Cultural Heritage. The architectural survey is one of the first actions taken in the knowledge process of an asset and, as such, it becomes essential for subsequent conservation, restoration and management operations. This paper describes an experience of integrated digital surveying techniques for the digitalization of an architecture by Andrea Palladio: Villa Forni Cerato. This experience has been carried out by the Heritage Survey Technology (HE.SU.TECH.) Group, from Politecnico di Milano - Polo di Mantova that has been carrying out research and consulting activities with institutions related to Italian Heritage since 2000. The case of Villa Forni Cerato is particularly interesting because, in this instance, digitization was integrated with other documental sources. The survey of the villa required the integration of all the different technologies used in the geomatic field for cultural HeritageThis case study, in particular, became an opportunity to test photogrammetric techniques with different photographic sets, acquired both from the ground and by means of Unmanned Aerial Systems (UAS). This work has been the subject of discussion within the exchange project between the two universities: Politecnico di Milano and Universidade de São Paulo.Este artículo trae un estudio en profundidad sobre documentación, representación y praxis para la conservación del patrimonio arquitectónico en el campo de la investigación académica aplicada. Centra-se en los avances tecnológicos en la documentación digital aplicada al Patrimonio Cultural. Se destaca la importancia de conciliar el conocimiento de la investigación académica con el trabajo técnico aplicado. El mapeo arquitectónico es una de las primeras acciones que se realizan en el proceso de conocimiento de un bien y, como tal, se vuelve fundamental para las operaciones posteriores de conservación, restauración y gestión. Este artículo presenta una experiencia de técnicas de mapeo digital integrado para la digitalización de la Villa Forni Cerato (Palladio). Esta experiencia fue llevada a cabo por el Grupo de Investigación Heritage Survey Technology (HE.SU.TECH.) Del Politecnico di Milano - Polo di Mantova, que lleva a cabo actividades de investigación y consultoría con instituciones vinculadas al patrimonio italiano desde 2000. El caso de Villa Forni Cerato es interesante porque la digitalización se ha integrado con otras fuentes documentales, como los planos originales y secciones de la Villa. La encuesta de Villa requirió la integración de muchas tecnologías diferentes utilizadas en el campo geomático del Patrimonio Cultural. Este caso de estúdio se convirtió en una oportunidad para probar técnicas fotogramétricas con diferentes conjuntos fotográficos, adquiridos tanto desde tierra como a través de Sistemas Aéreos no Tripulados (UAS). Este trabajo ha sido objeto de discusión dentro del proyecto de intercambio entre las universidades: Politecnico di Milano y Universidade de São Paulo

The Leech Nervous System: A Valuable Model to Study the Microglia Involvement in Regenerative Processes

Microglia are intrinsic components of the central nervous system (CNS). During pathologies in mammals, inflammatory processes implicate the resident microglia and the infiltration of blood cells including macrophages. Functions of microglia appear to be complex as they exhibit both neuroprotective and neurotoxic effects during neuropathological conditions in vivo and in vitro. The medicinal leech Hirudo medicinalis is a well-known model in neurobiology due to its ability to naturally repair its CNS following injury. Considering the low infiltration of blood cells in this process, the leech CNS is studied to specify the activation mechanisms of only resident microglial cells. The microglia recruitment is known to be essential for the usual sprouting of injured axons and does not require any other glial cells. The present review will describe the questions which are addressed to understand the nerve repair. They will discuss the implication of leech factors in the microglial accumulation, the identification of nerve cells producing these molecules, and the study of different microglial subsets. Those questions aim to better understand the mechanisms of microglial cell recruitment and their crosstalk with damaged neurons. The study of this dialog is necessary to elucidate the balance of the inflammation leading to the leech CNS repair

Cytokine Impregnated Biomatrix: A New Tool to Study Multi-Wall Carbon Nanotubes Effects on Invertebrate Immune Cells

The novel features of engineered nanoparticles, such as multi-wall carbon nanotubes (MWCNTs) are impressive and attractive for technology, however they dissolved in water and accumulate in soils through the application of sewage sludge, accidental spills, and deposition from the air, agrochemicals or soil remediation. Given that several studies have revealed that chronic exposure to these nanomaterials products through the ingestion of drinking water, inhalation and dermal contact may harbour potential risks to human health, risk assessment of this nanomaterials in the aquatic environment are becoming essential. Here we propose a freshwater invertebrate, the leech Hirudo medicinalis, as a model to assess the effects MWCNTs on the immune system by means of in vivo and in vitro experiments. For this study, we used a consolidated experimental approach based on injection in the body wall of the leech of the biomatrice Matrigel (MG), added with a specific macrophage chemoattractant, the cytokine Allograft inflammatory factor-1 (AIF-1) and/or with MWCNTs. MG sponges analysis show the presence of a larger number of cells positive for both CD68 and HmAIF-1, specific monocyte-macrophage markers. Ultrastructural analysis suggests that MWCNTs may be internalized by phagocytosis but they seem also to be able to pierce cell membranes during cells migration. Cells extracted from MG were also used for in vitro treatment with MWCNTs at different concentration (2.5, 5, 10, 25, 50 and 100 \ub5g/ml) for 24 h to study cell morphology changes and production of amyloid fibrils in order to encapsulate the foreign bodies. Our results, not only confirm the ability of MWCNTs in inducing a potent inflammatory response, but highlight rapid colorimetric assays that can be successfully used as sensitive tools for aquatic pollution biomonitoring