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Institute
- Biowissenschaften (2138) (remove)
Fuer die schlechte Prognose von Glioblastompatienten mit einer ueberlebenszeit von 9-15 Monaten (Norden and Wen, 2006) ist vor allem die hohe Invasivitaet dieser Tumore verantwortlich. Nach operativer Entfernung des Haupttumors entstehen aus den verbleibenden invadierten Zellen sekundaere Tumore, die sich mitunter ueber weite Bereiche des Hirns verteilen. Des Weitern sind die hochinvasiven Tumorzellen oft resistent gegen Chemo- und Strahlentherapie (Drappatz et al., 2009; Lefranc et al., 2005). In Maustumormodellen und Pateinten konnte zudem gezeigt werden, dass die neuartige antiangiogenetische Therapie zwar das Tumorwachstum verringert, jedoch die Invasivitaet stark erhoeht. (Norden et al., 2008; Ebos et al., 2009; Paez-Ribes et al., 2009). Ueber die Mechanismen die diese hohen Invasivitaet induzieren, ist bislang nur sehr wenig bekannt. Die durch Reduktion von Blutgefaessen steigende Hypoxie des Tumors foerdert die Expression von Matrix-Metalloproteinasen (MMPs). Dies fuehrt zum Abbau der extrazelluaeren Matrix des umgebenden gesunden Gewebes und beguenstigt dadurch die Tumorzellinvasion (Indelicato et al., 2010; Miyazaki et al., 2008; Shyu et al., 2007). Die Umformung des Aktinzytoskeletts und damit die Mobilitaet von Zellen wird vorwiegend durch ein akkurates Zusammenspeil der Rho GTPasen Rac, Rho und Cdc42, kontrolliert (Ridley et al., 2003). Fuer die Organisation von Axonen im Nervensystem und fuer die Blut- und Lymphgefaessbildung wurde gezeigt, dass die Interaktion der Eph-Rezeptortyrosinkinasen und Ihrer Ephrin-Liganden Signalwege induziert, die in die Regulation dieses Zusammenspiels involviert sind (Egea and Klein, 2007; Makinen et al., 2005; Palmer et al., 2002; Sawamiphak et al., 2010). Des Weiteren zeigt die Analyse der Genloci von Eph-Rezeptoren und Ephrinen in verschieden Hirntumoren eine gehaeufte Deletionen des Ephrin-B2-Gens. Die Quantifizierung von Ephrin-B2 mRNA in diesen Tumoren hat ausserdem ergeben, dass mit zunehmender Malignitaet die Expression von Ephrin-B2 sinkt. Aus diesen Gruenden wurden die Untersuchungen in dieser Arbeit auf die Rolle von Ephrin-B2 anhaengigen Signalwegen in der Glioblastomzellinvasion konzentriert. In einem modifiziertem Boyden-Chamber-Assay konnte gezeigt werden, dass das Ephrin-B2 induzierte EphB4 forward signaling und EphB4 induzierte Ephrin-B2 reverse signaling die Invasivitaet der human Glioblastomzelllinien LN-229, G55 und SNB-19 reduziert. In einem Maustumormodel konnte weiterhin gezeigt werden, dass Ephrin-B2 Knock-Out (KO) Astrozytomzellen, im Vergleich zu Wild-Typ (WT) Zellen, Tumore mit einem groesseren Volumen und einer erhoehten Invasivitaet bilden. Da die Expressionslevel fuer die Ephrin-B2 bindenden Rezeptoren EphA4, EphB1 EphB3 und EphB6 auch im adulten Hirn hoch sind (Hafner et al., 2004), weisen diese in vitro und in vivo Ergebnisse auf eine Tumorsupressorfunktion von Ephrin-B2 hin, die durch repulsive Effekte des Ephrin-B2 reverse signaling vermittelte werden koennten. Dies geht mit Erkenntnissen ueber kolorektale Tumore einher (Batlle et al., 2005). Die in einem Sphaeroid-Invasionsassay mit einer EphB-Rezeptoren freien Umgebung beobachtete verminderte Invasion von Ephrin-B2 WT deutet auf eine zusaetzliche invasionsblockierende Rolle der Ephrin-B2-Eph-Rezeptor Interaktion zwischen benachbarten Tumorzellen hin, wie sie auch in Brusttumoren gefunden wurde (Noren et al., 2006). Es scheint als sei Tumorprogression und Invasion erst moeglich, nachdem die Expression von Ephrin-B2 vermindert wurde. Es konnte weiterhin gezeigt werden, dass in hypoxischen Glioblastomzellen die Ephrin-B2 Expression durch die direkte Bindung des den Transkriptionsfaktors ZEB2 an den Ephrin-B2 Promoter reprimiert wird. In einem Weiteren Maustumormodel konnte gezeigt werden, dass die Blockierung der ZEB2 Expression mittels shRNA und die damit einhergehenden Inhibition der hypoxie induzierten Ephrin-B2 Repression das Wachstum und die Invasivitaet von Glioblastomen verringert. Zusaetzlich wurde gezeigt, dass der Verlust von ZEB2 ausreicht, die durch antiangiogenetische Therapie induzierte stark erhoehte Invasivitaet zu vermeiden. Die in dieser Arbeit gewonnen Erkenntnisse fuehren zu folgendem Modelmechanismus. In kleinen normoxischen Tumoren koennen repulsive Effekte des Ephrin-B2 reverse signalings und EphB forward signalings zwischen Tumorzellen und Zellen des umgebenden Gewebes die Ausbreitung und Invasion des Tumors unterdruecken. Zusaetzlich koennte das Ephrin-B2 induzierte EphB forward signaling zwischen benachbarten Tumorzellen die Mobilitaet der Tumorzellen wie in Brusttumoren inhibieren. Beim Erreichen einer bestimmten Tumorgroesse tritt Hypoxie auf, wodurch HIF-1alpha stabilisiert wird. Dies fuehrt dann zur ZEB2 Expression und leitet die Repression von Ephrin-B2 ein, was wiederum zur erhoehten Tumorzellemobilitaet und im Zusammenspiel mit MMPs zu Invasion fuehren kann. Gleichzeitig werden durch den HIF-induzierten VEGF-Gradienten neue Blutgefaesse rekrutiert. Damit wird der hypoxie-induzierten Invasivitaet entgegengewirkt. Wird mittels antiangiogenetischer Behandlung versucht Tumorprogression entgegenzuwirken, resultiert daraus eine erneut gesteigerte Hypoxie, die dann durch die ZEB2 vermittelte Repression von Ephrin-B2 wieder eine erhoehte Invasivitaet induzieren kann. Das Blockieren der ZEB2 Expression kann dieser durch antiangiogenetischen Behandlung induzierten Invasivitaet entgegenwirken.
Aim: To provide distribution information and preliminary conservation assessments for all species of the pineapple family (Bromeliaceae), one of the most diverse and ecologically important plant groups of the American tropics—a global biodiversity hotspot. Furthermore, we aim to analyse patterns of diversity, endemism and the conservation status of the Bromeliaceae on the continental level in the light of their evolutionary history.
Location: The Americas.
Methods: We compiled a dataset of occurrence records for 3,272 bromeliad species (93.4% of the family) and modelled their geographic distribution using either climate‐based species distribution models, convex hulls or geographic buffers dependent on the number of occurrences available. We then combined this data with information on taxonomy and used the ConR software for a preliminary assessment of the conservation status of all species following Criterion B of the International Union for the Conservation of Nature (IUCN).
Results: Our results stress the Atlantic Forest in eastern Brazil, the Andean slopes, Central America and the Guiana Highlands as centres of bromeliad diversity and endemism. Phylogenetically ancient subfamilies of bromeliads are centred in the Guiana highlands whereas the large radiations of the group spread across different habitats and large geographic area. A total of 81% of the evaluated bromeliad species are Possibly Threatened with extinction. We provide range polygons for 3,272 species, as well as newly georeferenced point localities for 911 species in the novel “bromeliad” r package, together with functions to generate diversity maps for individual taxonomic or functional groups.
Main conclusions: Diversity centres of the Bromeliaceae agreed with macroecological patterns of other plant and animal groups, but show some particular patterns related to the evolutionary origin of the family, especially ancient dispersal corridors. A staggering 2/3rds of Bromeliaceae species might be threatened with extinction, especially so in tropical rain forests, raising concerns about the conservation of the family and bromeliad‐dependent animal species.
PaCATB : a secreted catalase protecting Podospora anserina against exogenous oxidative stress
(2011)
A differential mass spectrometry analysis of secreted proteins from juvenile and senescentPodospora anserina cultures revealed age-related differences in protein profiles. Among other proteins with decreased abundance in the secretome of senescent cultures a catalase, termed PaCATB, was identified. Genetic modulation of the abundance of PaCATB identified differential effects on the phenotype of the corresponding strains. Deletion of PaCatB resulted in decreased resistance, over-expression in increased resistance against hydrogen peroxide. While the lifespan of the genetically modified strains was found to be unaffected under standard growth conditions, increased exogenous hydrogen peroxide stress in the growth medium markedly reduced the lifespan of the PaCatB deletion strain but extended the lifespan of PaCatB over-expressors. Overall our data identify a component of the secretome of P. anserina as a new effective factor to cope with environmental stress, stress that under natural conditions is constantly applied on organisms and influences aging processes.
Untersuchungen zur Bedeutung von Superoxid-Dismutasen für die Alterung von Podospora anserina
(2012)
Im Rahmen dieser vorliegenden Doktorarbeit sollte die Bedeutung von Superoxid-Dismutasen für das Resistenzverhalten und den Alterungsprozess bei P. anserina untersucht werden. Folgende Befunde aus den Analysen konnten erhalten werden:
1. Lokalisationsstudien der drei PaSods: Aus den biochemischen und fluoreszenzmikroskopischen Untersuchungen der drei verschiedenen PaSODs geht hervor, dass PaSOD1, eine Cu/ZnSOD, überwiegend im Cytosol und zu einem geringen Anteil im mitochondrialen Intermembranraum lokalisiert ist. Eine der beiden MnSODs, PaSOD2, wird vermutlich zur Abwehr von exogenem Superoxid sekretiert. Bei PaSOD3 handelt es sich um eine mitochondriale MnSOD.
2. Generierung von verschiedenen PaSod-Mutanten: Im Rahmen dieser Arbeit wurden von jeder PaSod mindestens drei unabhängige Überexpressionsstämme, ein GFP-Stamm- und ein Deletionsstamm hergestellt. Weiterhin wurden alle möglichen Doppel-Deletionsstämme und die Dreifach-Deletionsmutante erzeugt. Alle Stämme wurden auf DNA-Ebene verifiziert, zusätzlich wurde die Proteinmenge bzw. –Aktivität überprüft.
3. Einfluss der PaSODs auf die ROS-Toleranz: Die Analysen der ROS-Resistenzen haben gezeigt, dass PaSODs eine wichtige Rolle in der Entgiftung von Superoxiden spielt. So ließ sich bei den Deletionsstämmen der PaSods eine gesteigerte Sensitivität gegenüber Paraquat feststellen. Eine Aufsummierung der Sensitivität gegenüber Paraquat ist bei der PaSod-Tripelmutante (ΔPaSod1/2/3) zu erkennen.
Überraschenderweise kann durch die gesteigerten Mengen an aktiver PaSOD in den Überexpressionsstämmen (PaSod1-3_OEx) keine verbesserte Resistenz gegenüber Paraquat erzielt werden. Darüber hinaus führt die Überexpression des Gens für die mitochondriale SOD, PaSOD3, zu massiven negativen Effekten.
4. Einfluss auf die Lebensspanne: Durch eine fehlende Entgiftung von Superoxid in den PaSod-Deletionsmutanten ist eine Verminderung der Lebensspanne nicht festzustellen. Bei PaSod-Mutantenstämme, die eine erhöhte PaSOD-Aktivität und damit eine gesteigerte Abbaurate des Superoxids aufweisen, kann bei den PaSod1- und PaSod2-Überexpressionsstämmen keine verbesserte Lebensspanne unter den gewählten Standardbedingungen erzielt werden. Vielmehr noch ist die Lebensspanne der PaSod3-Überexpressionsstämme stark reduziert.
5. Einfluss der PaSod-Modulation auf andere Komponenten des ROS-Abbausystems: Die PaSOD-Aktivitäten scheinen miteinander co-reguliert zu werden. Des Weiteren scheint es ein Zusammenhang zwischen den beiden sekretierten Enzymen PaSOD2 und PaCATB zu geben. Deutlich wird auch, dass die Modulation der Superoxid-Dismutasen eine weitreichende Auswirkung auf andere Schutzsysteme hat. Beispielweise konnte gezeigt werden, dass Komponenten des mitochondrialen ROS-Schutzsystems und der Protein-Qualitätskontrolle in den PaSod3-Überexpressionsstämmen verändert sind.
Zusammenfassend lassen die Analysen der PaSod-modulierten Stämme den Schluss zu, dass die Superoxid-Dismutase in P. anserina ein wichtiges Enzym zum Abbau des schädlichen Superoxids darstellt, welches aber nur eine untergeordnete Rolle bei der Kontrolle der Lebensspanne unter den gewählten Wachstumsbedingungen im Labor ausübt. Des Weiteren haben die Analysen gezeigt, dass es durch die Modulation der PaSod-Gene zu weitreichenden Änderungen, die das ROS-Schutzsystem (PaSOD, PaCATB und PaPRX1) sowie die Protein-Qualitätskontrolle (PaHSP60, PaLON und PaCLPP) betreffen, kommt. Welche Auswirkung dabei diese Veränderungen in Bezug auf die Lebensspanne hat, kann nur schwer abgeschätzt werden und muss mit weiteren Untersuchungen geklärt werden.
Die Neurowissenschaften sind in Forschungsarbeiten für Schüler und Studierende immer wieder als eines der schwierigsten Teilgebiete der Biologie angeführt. Die Inhalte werden überwiegend nicht verstanden. Als mögliche Ursache gelten die seltenen praktischen Zugänge für die Lernenden aufgrund limitierter Ressourcen. Diese Ursache konnte in der vorliegenden Arbeit durch eine Befragung der Lehrkräfte zu ihren Praxisumsetzungen bestätigt werden. 70 % der Lehrkräfte gaben an, dass sie keine Experimente in der Schule zum Thema Nervenzellen anbieten. Experimente zur Verhaltensbiologie führen 65 % der Lehrkräfte nicht durch.
Um Schülern die Möglichkeit zu geben, sich experimentell mit den Themenfeldern der Neuro- und Verhaltensbiologie auseinanderzusetzen, wurden im Rahmen der vorliegenden Arbeit Schülerlabortage auf dem Feld der Neurowissenschaften konzipiert. Die Konzepte wurden schülerorientiert umgesetzt und neurowissenschaftliche Forschung durch den eigenen Umgang mit modernen Forschungsapparaturen erfahrbar gemacht. Die drei Labortage für die Sekundarstufe II wurden wissenschaftlich begleitet: 1) Verhaltensbiologie, 2) systemische Ebene der Elektrophysiologie, 3) elektrophysiologische Forschungsmethoden. Um die Qualität und Wirksamkeit der Labortage beurteilen zu können, wurden sie mit Feedbackerhebungen begleitet. Die drei Labortage wurden sowohl von den Lehrkräften als auch von den Schülern bezüglich ihrer Qualität positiv bewertet. Für die Schüler konnte gezeigt werden, dass die Beurteilung weitgehend unabhängig von einem zugrunde liegenden Interesse an Biologie und Forschung ausfällt. Anhand einer retrospektiven Erhebung wird außerdem gezeigt, dass alle drei Labortage eine höchst signifikante, selbsteingeschätzte Steigerung des „Wissens“, der „Anwendungszuversicht“ und des „Interesses“ bewirken. Schüler mit niedrigen Ausgangswerten zeigen einen besonders hohen Anstieg. Für das Interesse kann weiter gezeigt werden, dass auch Schüler mit hohem Ausgangswert eine große Interessenssteigerung durch den Labortag aufweisen. Das Interesse für den verhaltensbiologischen Labortag liegt etwas niedriger – die Labortage mit elektrophysiologischen Inhalten zeigen dagegen für die Anwendungszuversicht etwas niedrigere Werte.
Der Fokus der fachdidaktischen Forschung lag auf der Betrachtung des experimentellen Zugangs zur Elektrophysiologie über ein entwickeltes „EPhys-Setup“. Dabei handelt es sich um einen quasi-realen Messaufbau. Die Umsetzung kombiniert dazu Komponenten eines realen Elektrophysiologie-Setups (Hands-on Komponenten) mit einer speziell entwickelten schülerfreundlichen Software (Neurosimulation) und einem virtuellen Nervensystem in Form einer Platine. Als Modellnervensystem werden für diese Umsetzung Ganglien von Hirudo medicinalis verwendet – der Neurosimulation liegen originale elektrophysiologische Messspuren des Ganglions zugrunde. Experimentelle Vermittlungsansätze für die Elektrophysiologie finden sich kaum für den Schulbereich. Dem Bedarf einer entsprechenden Beforschung wurde mit verschiedenen Testinstrumenten nachgegangen, um den Vermittlungsansatz mit dem EPhys-Setup bewerten zu können. Dafür fand eine Wirksamkeitsanalyse über die Erhebung der Motivation der Schüler statt (Lab Motivation Scale; Dohn et al. 2016). Von Bedeutung war auch, inwiefern gegenüber der Umsetzung eine Technologieakzeptanz vorliegt (Technology Acceptance Model; Davis 1989), die im Schulkontext ausgehend von der steigenden Einbindung von Technologien einen entsprechenden Forschungsbedarf aufweist. Weiter wurde untersucht, ob sich die Bewertung des EPhys-Setups von der Bewertung einer Kontrollgruppe unterscheidet. Für die Kontrollgruppe wurde die Neurosimulation von den Hands-on Komponenten gelöst und die Schüler arbeiteten ausschließlich PC-basiert. Die Ergebnisse zeigen, dass beide Umsetzungen die Motivation förderten und eine Technologieakzeptanz bei den Schülern aufwiesen. Der Unterschied der Untersuchungsgruppen fällt gering aus. Die Abhängigkeiten, die für die verwendete Simulationsumsetzung gefunden wurden, beziehen sich ausschließlich auf Komponenten der „Freude“. Somit wird der intrinsische Bereich von den Schülern die am EPhys-Setup gearbeitet haben höher bewertet. Zur weiteren Analyse der Testinstrumente wurde auch eine Abhängigkeit der Bewertung vom zugrunde liegenden Biologieinteresse sowie von den Computerfähigkeiten vergleichend betrachtet. Der Einfluss auf die Bewertungen der drei Testskalen ist in vielen Fällen höher als der Einfluss der verwendeten Simulation. Vom individuellen Biologieinteresse der Schüler zeigen alle untersuchten Komponenten eine Abhängigkeit. Die größeren Effekte beziehen sich auf die Komponenten der „Lernwirksamkeit“ oder der „Freude“. Von den individuellen Computerfähigkeiten der Schüler zeigen Komponenten zur „Zuversicht bezüglich der Methoden und der Inhalte“ eine Abhängigkeit.
Highlights
• PUR, PVC and PLA microplastics affect life-history parameters of Daphnia magna.
• Natural kaolin particles are less toxic than microplastics.
• Microplastic toxicity is material-specific, e.g. PVC is most toxic on reproduction.
• In case of PVC, plastic chemicals are the main driver of microplastic toxicity.
• PLA bioplastics are similarly toxic as conventional plastics.
Abstract
Given the ubiquitous presence of microplastics in aquatic environments, an evaluation of their toxicity is essential. Microplastics are a heterogeneous set of materials that differ not only in particle properties, like size and shape, but also in chemical composition, including polymers, additives and side products. Thus far, it remains unknown whether the plastic chemicals or the particle itself are the driving factor for microplastic toxicity. To address this question, we exposed Daphnia magna for 21 days to irregular polyvinyl chloride (PVC), polyurethane (PUR) and polylactic acid (PLA) microplastics as well as to natural kaolin particles in high concentrations (10, 50, 100, 500 mg/L, ≤ 59 μm) and different exposure scenarios, including microplastics and microplastics without extractable chemicals as well as the extracted and migrating chemicals alone. All three microplastic types negatively affected the life-history of D. magna. However, this toxicity depended on the endpoint and the material. While PVC had the largest effect on reproduction, PLA reduced survival most effectively. The latter indicates that bio-based and biodegradable plastics can be as toxic as their conventional counterparts. The natural particle kaolin was less toxic than microplastics when comparing numerical concentrations. Importantly, the contribution of plastic chemicals to the toxicity was also plastic type-specific. While we can attribute effects of PVC to the chemicals used in the material, effects of PUR and PLA plastics were induced by the mere particle. Our study demonstrates that plastic chemicals can drive microplastic toxicity. This highlights the importance of considering the individual chemical composition of plastics when assessing their environmental risks. Our results suggest that less studied polymer types, like PVC and PUR, as well as bioplastics are of particular toxicological relevance and should get a higher priority in ecotoxicological studies.
Plastic products leach chemicals that induce in vitro toxicity under realistic use conditions
(2021)
Plastic products contain complex mixtures of extractable chemicals that can be toxic. However, humans and wildlife will only be exposed to plastic chemicals that are released under realistic conditions. Thus, we investigated the toxicological and chemical profiles leaching into water from 24 everyday plastic products covering eight polymer types. We performed migration experiments over 10 days at 40 °C and analyzed the migrates using four in vitro bioassays and nontarget high-resolution mass spectrometry (UPLC-QTOF-MSE). All migrates induced baseline toxicity, 22 an oxidative stress response, 13 antiandrogenicity, and one estrogenicity. Overall, between 17 and 8681 relevant chemical features were present in the migrates. In other words, between 1 and 88% of the plastic chemicals associated with one product were migrating. Further, we tentatively identified ∼8% of all detected features implying that most plastic chemicals remain unknown. While low-density polyethylene, polyvinyl chloride, and polyurethane induced most toxicological endpoints, a generalization for other materials is not possible. Our results demonstrate that plastic products readily leach many more chemicals than previously known, some of which are toxic in vitro. This highlights that humans are exposed to many more plastic chemicals than currently considered in public health science and policies.
Plastics contain a complex mixture of chemicals including polymers, additives, starting substances and side-products of processing. These plastic chemicals are prone to leach into the packaged goods, in the case of food contact materials (FCMs), or into the natural environment, in the case of plastic debris. Thus, plastics represent an exposure source of chemicals for humans and wildlife alike. While it is widely known that individual plastic chemicals, such as bisphenol A and phthalates, are hazardous, little is known on the overall chemical composition and toxicity of plastics. When fragmented into smaller particles, referred to as microplastics (< 5 mm), the plastic itself can be ingested by many species. It is well established that microplastic ingestion can have negative consequences for a wide range of organisms including invertebrates, but the contribution of plastic chemicals to the toxicity of microplastics is unclear.
Given the above, the present thesis aimed at a comprehensive toxicological, ecotoxicological and chemical characterization of everyday plastics. For a comparative evaluation, 77 plastic products were selected covering 16 material types (e.g., polyethylene) made from petroleum or renewable feedstocks. These products included biodegradable products, FCMs and non-FCMs, as well as raw materials and final products, respectively. In the first two studies, the chemical mixtures contained in the 77 products were extracted with methanol and extracts were analyzed in a set of four in vitro bioassays and by non-target high-resolution gas or liquid chromatography mass spectrometry. Since an exposure only occurs if chemicals actually leach under realistic conditions, in a third study migration experiments with water were conducted for 24 out of the 77 products. The aqueous migrates were assessed in the same way as the methanolic extracts. In addition, the freshwater invertebrate Daphnia magna was exposed chronically to microplastics made of polyvinylchloride (PVC), polyurethane (PUR) and polylactic acid (PLA) to investigate the contribution of chemicals in microplastic toxicity, in a fourth study.
The experimental findings demonstrate that a wide variety of chemicals is present in plastics. A single plastic product can contain up to several thousand chemical features, most of which unique to that product and at the same time unknown. The results also indicate that the majority of these chemical mixtures are toxic in vitro. Accordingly, 65% of the plastic extracts induced baseline toxicity and 42% an oxidative stress response, while 25% had an antiandrogenic and 6% an estrogenic activity. This implies that chemicals causing unspecific toxicity are more prevalent in plastics than such with endocrine effects. These chemicals can also leach from plastics under realistic conditions. Between 17 and 8936 chemical features were detected in a single migrate sample and all 24 tested migrates induced in vitro toxicity. This means that humans and wildlife can actually be exposed to toxic plastic chemicals under realistic conditions. Generally, each product has its individual toxicological and chemical fingerprint. Thus, neither material type, feedstock, biodegradability nor the food contact suitability of a product can serve as a predictor for the toxicity, the chemical composition or complexity of a product. Likewise, this means that bio-based and biodegradable materials are not superior to their petroleum-based counterparts from a toxicological perspective despite being promoted as sustainable alternatives to conventional plastics.
Moreover, the present thesis demonstrates that plastic chemicals can be the main driver for microplastic toxicity. Irregular microplastics made of PVC, PUR and PLA adversely affected life-history traits of D. magna in a polymer type- and endpoint-dependent manner at concentrations between 100 and 500 mg L-1 and with a higher efficiency than natural kaolin particles. While the toxicity of PVC was triggered by the chemicals used in the material, the effects of PUR and PLA were induced by the physical properties of the particle.
In addition, in the fifth study, results and observations made during this thesis were integrated inter- and transdisciplinarily with the perspectives of a social scientist and a product manufacturer. This elucidated that knowledge on plastic ingredients is often concealed, is lacking or not applicable in practice. These intransparencies hinder the safety evaluation of plastic products as well as the choice and sale of the least toxic packaging material.
Overall, the present thesis highlights that the chemical safety of plastics and their bio-based and biodegradable alternatives is currently not ensured. Thus, chemicals require more consideration in the toxicity and risk assessment of plastics and microplastics. Product-specific and complex chemical compositions, including unknown compounds, pose a challenge here. Two essential steps towards non-toxic products are to increase transparency along the product life cycle and to reduce the chemical complexity of plastics by communication and regulation. The results of the present thesis indicate that products exist which do not contain toxic chemicals. These can serve to direct the design of safer plastics. Since toxicity and chemical complexity seem to increase with processing, the integration of toxicity testing during the production steps would further support the safe and sustainable production and use of plastic products.
Geoffrey Burnstock will be remembered as the scientist who set up an entirely new field of intercellular communication, signaling via nucleotides. The signaling cascades involved in purinergic signaling include intracellular storage of nucleotides, nucleotide release, extracellular hydrolysis, and the effect of the released compounds or their hydrolysis products on target tissues via specific receptor systems. In this context ectonucleotidases play several roles. They inactivate released and physiologically active nucleotides, produce physiologically active hydrolysis products, and facilitate nucleoside recycling. This review briefly highlights the development of our knowledge of two types of enzymes involved in extracellular nucleotide hydrolysis and thus purinergic signaling, the ectonucleoside triphosphate diphosphohydrolases, and ecto-5′-nucleotidase.