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Blowflies --- Calliphoridae --- Diptera --- Entomologia Medica E Veterinaria --- Flies --- Insecta. Diptera (Sistematica) --- North America

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Het werk voorgesteld in deze thesis concentreert zich rond de identificatie van fysiologische processen waarbij neprilysine-achtige genen van Drosophila melanogaster betrokken zijn. Deze enzymen behoren tot de familie van M13 metalloproteasen waarvan neprilysine het best gekarakteriseerd is bij vertebraten. Deze klasse van enzymen vormt een belangrijk doelwit voor geneesmiddelen omwille van hun bijdrage in de ontwikkeling van cardiovasculaire ziekten, kanker en de ziekte van Alzheimer. De identificatie van deze enzymen in invertebraten biedt de mogelijkheid om nieuwe functies te karakteriseren die mogelijk geconserveerd zijn bij vertebraten.We hebben de expressie patronen van Drosophila Nep1-Nep5 genen doorheen de ontwikkeling bepaald met behulp van in situ hybridisatie. De expressie van deze enzymen in een brede waaier van weefsels en structuren toont hun betrokkenheid aan in talrijke fysiologische processen. Het uitvoeren van testen voor leren en geheugen op larven met verhoogde Nep1 expressie in de mushroom bodies toont aan dat neprilysine misschien betrokken is bij geheugenvorming in vliegen. Experimenten die de lokalisatie van Nep1 antilichaam kleuringen aantonen in de cellen die insuline-achtige peptiden secreteren suggereren dat Nep1 betrokken kan zijn bij insuline-signalisatie in Drosophila. Een ander interessant resultaat dat we hier beschrijven is de rol van Nep2 in celmigratie. De expressie van Nep2 in ovariële ‘border’ cellen suggereerde een rol voor Nep2 in de migratie van deze cellen tijdens de oögenese. Gerichte overexpressie en RNAi van Nep2 in deze migrerende cellen resulteerde respectievelijk in vertraagde delaminatie en migratie van de ‘border’ cellen. We tonen ook aan dat Nep2 en misschien Nep4 betrokken zijn bij de ontwikkeling van het embryonaal tracheeënsysteem. Embryo’s van kandidaat mutante allelen voor deze twee enzymen vertonen fusie en vertakkingsdefecten tijdens de morfogenese van de tracheeën. De verschillen in fenotypes veroorzaakt door de mutante allelen suggereren dat Nep2 en Nep4 werkzaam zijn in verschillende processen die belangrijk zijn voor de ontwikkeling van het tracheeënsysteem. De expressie van elk van de vijf neprilysines in de mannelijke voortplantingsorganen suggereert het belang van deze klasse van enzymen voor de spermatogenese en de mannelijke vruchtbaarheid zoals reeds beschreven is bij zoogdieren. The work presented in this thesis focuses on the identification of physiological pathways in which neprilysin-like enzymes in Drosophila melanogaster are involved. These enzymes belong to the family of M13 metalloproteases, of which neprilysin is the best characterized member in vertebrates. This class of enzymes represents important drug targets due to their contribution in the development of cardiovascular diseases, cancer and Alzheimer’s disease. Identification of these enzymes in invertebrate species offers opportunities to characterize new functions which might be conserved in vertebrates.We studied the gene expression patterns of Drosophila Nep1-Nep5 by in situ hybridization throughout development. The wide range of tissues and structures in which these enzymes are expressed suggests that they are involved in numerous physiological pathways. By performing olfactory learning tests on larvae with Nep1 overexpression in the mushroom bodies we showed that Nep1 might be involved in the process of learning and memory in flies. Localization of Nep1 antibody staining in the insulin-like peptide producing cells suggests that Nep1 may be involved in insulin signaling in Drosophila. Another interesting result we report here is the role of Nep2 in cell migration. The expression of Nep2 in ovarian border cells suggested a role of Nep2 in the migration of these cells during oogenesis. By targeting overexpression and RNAi in these migrating cells we observed delayed delamination and migration of the border cells respectively. We also show that Nep2 and maybe Nep4 are involved in the development of the embryonic tracheal system. Embryos of candidate mutant alleles for these two enzymes reveal fusion and branching defects during tracheal branching morphogenesis. Based on the differences between phenotypes produced by the alleles we suggest that Nep2 and Nep4 are implicated in different processes that contribute to the development of the tracheal system. Expression of all five neprilysins in the male reproductive organs suggests the significance of this class of enzymes in spermatogenesis and male fertility as is described for their mammalian counterparts.

Academic collection --- 577.2 --- 577.15 --- 595.772.4 --- 595.772.4 Schizophora. Calliphoridae. Drosophilidae. Muscidae. Sarcophagidae. Trypetidae. Glossina (tsetse flies). Anthomyiidae. Psilidae. Oestridae. Agromyzidae. Tachinidae --- Schizophora. Calliphoridae. Drosophilidae. Muscidae. Sarcophagidae. Trypetidae. Glossina (tsetse flies). Anthomyiidae. Psilidae. Oestridae. Agromyzidae. Tachinidae --- 577.15 Enzymes. Catalysts of biological reactions. Enzymology --- Enzymes. Catalysts of biological reactions. Enzymology --- 577.2 Molecular bases of life. Molecular biology --- Molecular bases of life. Molecular biology --- Theses

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In 2001, Benecke concluded a review on the history of forensic entomology with these optimistic words: "basic research and advanced application of forensic entomology (…) has opened the way to routine casework". At the same time, the TV show Crime Scene Investigation (CSI) largely brought forensic entomology to light. However, the show also cruelly pointed out its limits: After the team leader explained to his colleague how insects can help determine the time of death, the team leader added "You've still got to convince a jury", to which the colleague promptly responded "On guns. It's got to be better than bugs. Less Latin." Indeed, several factors—including complexity, inherent limitations, and the rapid evolution of scientific knowledge—explain the slow acceptance of insect-based evidence. In this context, this Special Issue focuses on the articulation between laboratory studies and casework, a major challenge for the future of forensic entomology.

Psychology --- Diptera --- identification --- forensic entomology --- funerary archaeoentomology --- crime scene --- autopsy --- cooling period --- entomological evidence --- expertise --- casework --- court --- criminal justice systems --- expert witness --- insect evidence --- research --- postmortem interval --- development --- succession --- species identification --- animal carcass --- cadaver --- decaying substrate --- insect succession --- successional studies --- vertebrate decomposition --- animal carcasses --- bait attraction --- ADD --- TBS --- PMI --- colonisation --- temperature --- medico-legal entomology --- time of colonization --- accumulated degree day estimates --- length-weight estimates --- species interactions --- Calliphoridae --- legislation --- expert witness statement --- criteria --- limitations --- thanatology --- confession --- post-mortem interval --- carrion --- larva --- first record --- barcoding DNA --- integrative taxonomy --- arthropods --- burial --- decay --- insects --- pig --- biological variation --- death time estimation --- alternative storage --- carrion insects --- validation --- minimum postmortem interval (PMI-min) --- rearing --- calliphoridae --- Lucilia sericata --- climate change --- global warming --- Fanniidae --- larval morphology --- human cadaver --- Forensic Entomology --- Spain --- experimental studies --- cases --- cold cases --- hair evidence --- n/a

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In 2001, Benecke concluded a review on the history of forensic entomology with these optimistic words: "basic research and advanced application of forensic entomology (…) has opened the way to routine casework". At the same time, the TV show Crime Scene Investigation (CSI) largely brought forensic entomology to light. However, the show also cruelly pointed out its limits: After the team leader explained to his colleague how insects can help determine the time of death, the team leader added "You've still got to convince a jury", to which the colleague promptly responded "On guns. It's got to be better than bugs. Less Latin." Indeed, several factors—including complexity, inherent limitations, and the rapid evolution of scientific knowledge—explain the slow acceptance of insect-based evidence. In this context, this Special Issue focuses on the articulation between laboratory studies and casework, a major challenge for the future of forensic entomology.

Psychology --- Diptera --- identification --- forensic entomology --- funerary archaeoentomology --- crime scene --- autopsy --- cooling period --- entomological evidence --- expertise --- casework --- court --- criminal justice systems --- expert witness --- insect evidence --- research --- postmortem interval --- development --- succession --- species identification --- animal carcass --- cadaver --- decaying substrate --- insect succession --- successional studies --- vertebrate decomposition --- animal carcasses --- bait attraction --- ADD --- TBS --- PMI --- colonisation --- temperature --- medico-legal entomology --- time of colonization --- accumulated degree day estimates --- length-weight estimates --- species interactions --- Calliphoridae --- legislation --- expert witness statement --- criteria --- limitations --- thanatology --- confession --- post-mortem interval --- carrion --- larva --- first record --- barcoding DNA --- integrative taxonomy --- arthropods --- burial --- decay --- insects --- pig --- biological variation --- death time estimation --- alternative storage --- carrion insects --- validation --- minimum postmortem interval (PMI-min) --- rearing --- calliphoridae --- Lucilia sericata --- climate change --- global warming --- Fanniidae --- larval morphology --- human cadaver --- Forensic Entomology --- Spain --- experimental studies --- cases --- cold cases --- hair evidence