Refine
Has Fulltext
- no (56) (remove)
Year of publication
- 2023 (56) (remove)
Document Type
- Doctoral Thesis (56) (remove)
Language
- English (56) (remove)
Is part of the Bibliography
- yes (56)
Keywords
- photosynthesis (3)
- Arabidopsis thaliana (2)
- Photosynthese (2)
- fluctuating light (2)
- metabolomics (2)
- transcriptomics (2)
- (Xeno)Hormone (1)
- (xeno)hormones (1)
- Affordances (1)
- African weakly electric fish (1)
Institute
- Institut für Biochemie und Biologie (28)
- Institut für Physik und Astronomie (8)
- Institut für Chemie (4)
- Extern (2)
- Fachgruppe Betriebswirtschaftslehre (2)
- Institut für Ernährungswissenschaft (2)
- Institut für Geowissenschaften (2)
- Department Psychologie (1)
- Department Sport- und Gesundheitswissenschaften (1)
- Fachgruppe Politik- & Verwaltungswissenschaft (1)
This book provides empirical evidence that all States have a universally binding obligation to adopt national laws and international treaties to protect the marine environment, including the designation of Marine Protected Areas. Chapter by chapter this obligation is detailed, providing the foundation for holding States responsible for fulfilling this obligation. The fundamentals are analysed in a preliminary chapter, which examines the legally binding sources of the Law of the Sea as well as its historical development to help readers understand the key principles at hand.
The Law of the Sea provides more than 1000 instruments and more than 300 regulations concerning marine protection. While the scope of most treaties is limited either regarding species, regions or activities, one regulation addresses States in all waters: the obligation to protect and preserve the marine environment as stipulated under Art. 192 of the 1982 United Nations Convention on the Law of the Sea (UNCLOS). As this ‘Constitution of the Ocean’ not only contains conventional laws but also very broadly reflects pre-existing rules of customary international law, an extensive analysis of all statements made by States in the UN General Assembly, their practices, national laws and regulations as well as other public testimonials demonstrates that Art. 192 UNCLOS indeed binds the whole community of States as a rule of customary international law with an erga omnes effect. Due to the lack of any objections and its fundamental value for humankind, this regulation can also be considered a new peremptory norm of international law (ius cogens).
While the sovereign equality of States recognises States’ freedom to decide if and how to enter into a given obligation, States can also waive this freedom. If States accepted a legally binding obligation, they are thus bound to it. Concerning the specific content of Art. 192 UNCLOS, a methodical interpretation concludes that only the adoption of legislative measures (national laws and international agreements) suffices to comply with the obligation to protect and preserve the marine environment, which is confirmed by the States’ practices and relevant jurisprudence. When applied to a specific geographical area, legislative measures to protect the marine environment concur with the definition of Marine Protected Areas. Nonetheless, as the obligation applies to all waters, the Grotian principle of the freedom of the sea dictates that the restriction of activities through the designation of Marine Protected Areas, on the one hand, must be weighed against the freedoms of other States on the other. To anticipate the result: while all other rights under the UNCLOS are subject to and contingent on other regulations of the UNCLOS and international law, only the obligation to protect and preserve the marine environment is granted absolutely – and thus outweighs all other interests
Photosynthesis converts light into metabolic energy which fuels plant growth. In nature, many factors influence light availability for photosynthesis on different time scales, from shading by leaves within seconds up to seasonal changes over months. Variability of light energy supply for photosynthesis can limit a plant´s biomass accumulation. Plants have evolved multiple strategies to cope with strongly fluctuation light (FL). These range from long-term optimization of leaf morphology and physiology and levels of pigments and proteins in a process called light acclimation, to rapid changes in protein activity within seconds. Therefore, uncovering how plants deal with FL on different time scales may provide key ideas for improving crop yield. Photosynthesis is not an isolated process but tightly integrates with metabolism through mutual regulatory interactions. We thus require mechanistic understanding of how long-term light acclimation shapes both, dynamic photosynthesis and its interactions with downstream metabolism. To approach this, we analyzed the influence of growth light on i) the function of known rapid photosynthesis regulators KEA3 and VCCN1 in dynamic photosynthesis (Chapter 2-3) and ii) the interconnection of photosynthesis with photorespiration (PR; Chapter 4).
We approached topic (i) by quantifying the effect of different growth light regimes on photosynthesis and photoprotection by using kea3 and vccn1 mutants. Firstly, we found that, besides photosynthetic capacity, the activities of VCCN1 and KEA3 during a sudden high light phase also correlated with growth light intensity. This finding suggests regulation of both proteins by the capacity of downstream metabolism. Secondly, we showed that KEA3 accelerated photoprotective non-photochemical quenching (NPQ) kinetics in two ways: Directly via downregulating the lumen proton concentration and thereby de-activating pH-dependent NPQ, and indirectly via suppressing accumulation of the photoprotective pigment zeaxanthin.
For topic (ii), we analyzed the role of PR, a process which recycles a toxic byproduct of the carbon fixation reactions, in metabolic flexibility in a dynamically changing light environment. For this we employed the mutants hpr1 and ggt1 with a partial block in PR. We characterized the function of PR during light acclimation by tracking molecular and physiological changes of the two mutants. Our data, in contrast to previous reports, disprove a generally stronger physiological relevance of PR under dynamic light conditions. Additionally, the two different mutants showed pronounced and distinct metabolic changes during acclimation to a condition inducing higher photosynthetic activity. This underlines that PR cannot be regarded purely as a cyclic detoxification pathway for 2PG. Instead, PR is highly interconnected with plant metabolism, with GGT1 and HPR1 representing distinct metabolic modulators.
In summary, the presented work provides further insight into how energetic and metabolic flexibility is ensured by short-term regulators and PR during long-term light acclimation.
Unveiling the Local Universe
(2023)
The digitization has permeated almost all aspects of an individual’s life. In the work context as well as in the private sphere, one readily encounters and relies on Information and Communication Technologies (ICTs), such as Social Networking Sites (SNS), smartphones and so forth. By communicating with as well as obtaining information via such technologies, ICTs engage one’s mind as interaction happens. This interaction of ICTs and the human mind form the focal topic of this thesis. Within this thesis, the human mind is represented on behalf of a facilitated model comprising a perceptual, a cognitive and a motor subsystem. ICTs represent an external stimulus, which triggers the human mind's perceptual subsystem, the cognitive subsystem and eventually leads to a motoric response via the motor subsystem. The external stimulus causing this event chain is within this thesis an ICT. The digital environment and related ICTs are high attention environments offering large and easily accessible amounts of information. Not surprisingly, issues may arise, when the human mind deals with ICTs. Thus, the interplay between ICTs and the human mind entails downsides. This thesis investigates these downsides and in addition the ICT-based factors that cause these downsides. More specifically, the thesis investigates these two aspects as research questions in the context of SNSs as well as other ICTs (such as smartphones, e-learning etc.). Addressing the research questions, 8 articles are submitted within this thesis which address the topic with different methodologies, including quantitative, qualitative, mixed methods as well as systematic literature reviews. Article 1 investigates factors that lead to SNS fatigue and discontinuance intentions in a mixed-methods design. Article 2 explores if certain factors encountered on a newsfeed hamper sensemaking. Article 3 proposes a study design to explore the link between disorderly perceptions of a SNSs newsfeed and gender stereotype activations. Article 4 considers the interplay between users and algorithms via the newsfeed interface and the implications for relevance perceptions. Article 5 explores information acquisition, hampering factors and verification strategies of social media users. Article 6 systematically reviews addiction scales of various ICTs. Article 7 investigates click behavior in e-learning contexts and how this is linked to cultural and personality traits. Finally, article 8 offers a comprehensive overview of the antecedents and consequences of children’s smartphone usage. Within the specific context of SNSs, the thesis suggests that the cognitive tolls imposed on users’ minds cause adverse effects, such as impaired sensemaking, fatigue, stereotype activation as well as intentions to discontinue the service. Other ICTs lead to addiction, and i.e., smartphones evidence to cause cognitive impairments in children. Factors on the ICT side that promote these adverse effects are linked to specific features, such as the newsfeed for SNSs and entail overload or perceptions of disorder. The thesis adds theoretically to the understanding of downsides that arise from the interplay between human minds and ICTs. Especially, the context of SNSs is spotlighted and insights add to the growing body of literature on experiences and perceptions. For instance, one study’s result suggests that considering information organization is as important as merely decreasing overload perceptions from the users in countering adverse effects of SNS usage. Practically, the thesis emphasizes the importance of mindful interaction with ICTs. Future research is welcome to build on the exploratory investigations and may draw an even more holistic picture to enhance the interaction between ICTs and the human mind.
Functional materials, also called "Smart Materials", are described by their ability to fulfill a desired task through targeted interaction with its environment. Due to this functional integration, such materials are of increased interest, especially in areas where the increasing micronization of components is required. Modern manufacturing processes (e.g. microfluidics) and the availability of a wide variety of functional materials (e.g. shape memory materials) now enable the production of particle-based switching components. This category includes micropumps and microvalves, whose basic function is the active control of liquid flows. One approach in realizing those microcomponents as pursued by this work, enables variable size-switching of water-filled microballoons by implementing a stimulus-sensitive switching motif in the capsule's membrane shell, while being under the influence of a constant driving force. The switching motif with its gatekeeper function has a critical influence on one or more material parameters, which modulate the capsule's resistance against the driving force in microballoon expansion process. The advantage of this concept is that even non-variable analyte conditions, such as concentration levels of ions, can be capitalized to generate external force fields that, under the control of the membrane, cause an inflation of the microballoon by an osmotically driven water influx. In case of osmotic pressure gradients as the driving force for the capsule expansion, material parameters associated with the gatekeeper function are specifically the permeability and the mechanical stiffness of the shell material. While a modulation of the shell permeability could be utilized to kinetically impede the water influx on large time scales, a modulation of the shell's mechanical stiffness even might be utilized to completely prevent the capsule inflation due to a possible non-deformability beneath a certain threshold pressure. In polymer networks, which are a suitable material class for the demanded capsule shell because of their excellent elasticity, both the permeability and the mechanical properties are strongly influenced by the crystallinity of the material. Since the permeability is effectively reduced with increasing crystallinity, while the mechanical stiffness is simultaneously greatly increased, both effects point in the same direction in terms of their functional relationship. For this reason and due to a reversible and contactless modulation of the membrane crystallinity by heat input, crystallites may be suitable switching motifs for controlling the capsule expansion. As second design element of reversible expandable microballoons, the capsule geometry, defined by an aqueous core enveloped by the temperature-sensitive polymer network membrane, should allow an osmotic pressure gradient across the membrane layer. The strength of the inflation pressure and the associated inflation velocity upon membrane melting should be controlled by the salt concentration within the aqueous core, while a turn in the osmotic gradient should furthermore allow the reversible process of capsule deflation. Therefore, it should be possible to build either microvalves and micropumps, while their intended action of either pumping or valving is determined by their state of expansion and the direction of the osmotic pressure gradient.. Microballoons of approximately 300 µm in diameter were formed via droplet-based microfluidics from double-emulsion templates (w/o/w). The elastomeric capsule membrane was formed by photo-crosslinking of methacrylate (MA) functionalized oligo(ε-caprolactone) precursors (≈ 3.8 MA-arms, Mn ≈ 12000 g mol-1) within the organic medium layer (o) via UV-exposure after droplet-formation. After removal of the toluene/chloroform mixture by slow extraction via the continuous aqueous phase, the capsules solidified under the development of a characteristic "mushroom"-like shape at specific experimental conditions (e.g. λ = 308 nm, 57 mJ·s-1·cm-2, 16 min). It could be furthermore shown that in dependency to the process parameters: oligomer concentration and curing-time also spherical capsules were accessible. Long curing-times and high oligomer concentrations at a fixed light-intensity favored the formation of "mushroom"-like capsules, whereas the contrary led to spherical shaped capsules. A comparative study on thin polymer network films of same composition and equal treatment proved a correlation between the film's crosslink density and their contraction capability, while stronger crosslinked polymer networks showed a stronger contraction after solvent removal. In combination with observations during capsule solidification via light-microscopy, where a continuous shaping from almost spherical crosslinked templates to "mushroom"-shaped and solidified capsules was stated, the following mechanism was proposed. In case of low oligomer contents and short curing-times, the contraction of the capsule shell during solvent removal is strongly diminished due to a low degree of crosslinking. Therefore, the solidifying shell could freely collapse onto the aqueous core. In the other case, high oligomer concentrations and long curing-times will favor the formation of highly crosslinked capsule membranes with a strong contraction capability. Due to an observed decentered location of the aqueous core within the swollen polymer network, an uneven radial stress along the capsule's circumference is exerted to the incompressible core. This lead to an uneven contraction during solvent removal and a directed flow of the core fluid into the direction of the minimal stress vector. In consequence, the initially thicker spherical cap contracts, whereas the opposing thinner spherical cap get stretched. The "mushroom"-shape over some advantages over their spherical shaped counterparts, why they were selected for the further experiments. Besides the necessity of a high density of crosslinking for the purpose of extraordinary elasticity and toughness, the form-anisotropy promotes a faster microballoon expandability due to a partial reduction of the membrane thickness. Additionally, pre-stretched regions of thin thickness might provide a better resistance against inflation pressure than spherical but non-stretched capsules of equal membrane thickness. The resulting "mushroom"-shaped microcapsules exhibited a melting point of Tm ≈ 50 - 60 °C and a degree of crystallinity of Xc ≈ 29 - 38 % depending on the membrane thickness and internal salt content, which is slightly lower than for the non-crosslinked oligomer and reasoned by a limited chain mobility upon crosslinking. Nonetheless, the melting transition of the polymer network was associated with a strong drop in its mechanical stiffness, which was shown to have a strong influence on the osmotic driven expansion of the microcapsules. Capsules that were subjected to osmotic pressures between 1.5 and 4.7 MPa did not expand if the temperature was well below the melting point of the capsule's membrane, i.e. at room temperature. In contrast, a continuous expansion, while approaching asymptotically to a final capsule size, was observed if the temperature exceeded the melting point, i.e. 60 °C. Microballoons, which were kept for 56 days at ∆Π = 1.5 MPa and room temperature, did not change significantly in diameter, why the impact of the mechanical stiffness on the expansion behavior is considered to be the greater than the influence of the shell permeability. The time-resolved expansion behavior of the microballoons above their Tm was subsequently modeled, using difusion equations that were corrected for shape anisotropy and elastic restoring forces. A shape-related and expansion dependent pre-factor was used to dynamically address the influence of the shell thickness differences along the circumference on the inflation velocity, whereas the microballoon's elastic contraction upon inflation was rendered by the inclusion of a hyperelastic constitutive model. An important finding resulting from this model was the pronounced increase in inflation velocity compared to hypothetical capsules with a homogeneous shell thickness, which stresses the benefit of employing shape anisotropic balloon-like capsules in this study. Furthermore, the model was able to predict the finite expandability on basis of entropy-elastic recovery forces and strain-hardening effects. A comparison of six different microballoons with different shell thicknesses and internal salt contents showed the linear relationship between the volumetric expansion, the shell thickness and the applied osmotic pressure, as represented by the model. As the proposed model facilitates the prediction of the expansion kinetics depending on the membranes mechanical and diffusional characteristics, it might be a screening tool for future material selections. In course of the microballoon expansion process, capsules of intermediate diameters could be isolated by recrystallization of the membrane, which is mainly caused by a restoration of the membrane's mechanical stiffness and is otherwise difficult to achieve with other stimuli-sensitive systems. The capsule's crystallinity of intermediate expansion states was nearly unchanged, whereas the lamellar crystal size tends to decreased with the expansion ratio. Therefore, it was assumed that the elastic modulus was only minimally altered and might increased due to the networks segment-chain extension. In addition to the volume increase achieved by inflation, a turn in the osmotic gradient also facilitated the reversible deflation, which was shown in inflation/deflation cycles. These both characteristics of the introduced microballoons are important parameter regarding the realization of micropumps and microvalves. The fixation of expanded microcapsules via recrystallization enabled the storage of entropy-elastic strain-energy, which could be utilized for pumping actions in non-aqueous media. Here, the pumping velocity depended on both, the type of surrounding medium and the applied temperature. Surrounding media that supported the fast transport of pumped liquid showed an accelerated deflation, while high temperatures further accelerate the pumping velocity. Very fast rejection of the incorporated payload was furthermore realized with pierced expanded microballoons, which were subjected to temperatures above their Tm. The possible fixation of intermediate particle sizes provide opportunities for vent constructions that allowed the precise adjustment of specific flow-rates and multiple valve openings and closings. A valve construction was realized by the insertion of a single or multiple microballoons in a microfluidic channel. A complete and a partial closing of the microballoon-valves was demonstrated as a function of the heating period. In this context, a difference between the inflation and deflation velocity was stated, summarizing slower expansion kinetics. Overall, microballoons, which presented both on-demand pumping and reversible valving by a temperature-triggered change in the capsule's volume, might be suitable components that help to design fully integrated LOC devices, due to the implementation of the control switch and controllable inflation/deflation kinetics. In comparison to other state of the art stimuli-sensitive materials, one has to highlight the microballoons capability of stabilizing almost continuously intermediate capsule sizes by simple recrystallization of the microballoon's membrane.
Negotiations are a way of joint decision-making and thereby a form of social conflict. By determining the concrete allocation of scarce resources, negotiations have a great impact on the value creation of companies. If companies succeed in achieving better negotiation results in the long term, they can increase their profitability. Ensuring a company's negotiation success is therefore an organizational issue of central importance. While the question of ensuring individual negotiation success has been the subject and topic of multidisciplinary research for a long time, the question of how organizations can implement and ensure continuous negotiation success remains largely unexplored. This dissertation therefore aims to investigate how companies enable their employees to consistently achieve better negotiation outcomes. It is significant that, in the corporate context, negotiators do not act as individuals but as embedded representatives of an organization, and that negotiations are not one-time events but recurring necessities for the existence of the organization instead. In organizations, those recurring processes with a similar fundamental structure are handled by routines. A planned improvement of routines is often forced by new artifacts. In this context, artifacts refer to human-created technologies with which humans interact within routines and therefore artifacts have a central influence on executing the routine. If negotiation activities in companies are represented by organizational routines, one central issue for improving companies’ negotiation performance is the artifacts’ incorporation into organizational negotiation routines that facilitate the efficient application of the insights from negotiation research. The dissertation consists of three studies that were written as research papers to examine artifacts in the organizational negotiation context. The first study focuses on the pre-negotiation stage and presents four tools to assist negotiation practitioners in efficiently preparing for negotiation. The study examines the negotiation preparation’s effectiveness and efficiency and the negotiation outcome in a case-based experiment. The second study is devoted to a closer examination of the barriers that inhibit the adoption of negotiation support systems (NSSs) as one kind of organizational negotiation artifact. The investigation is conducted using a structural equation model based on information from participating practitioners. The third study is concerned with the future of negotiation support system research. An exploratory study based on qualitative in-depth interviews with proven and published experts in the field aims to evaluate the current state of research. The general discussion of the dissertation connects, summarizes, and concludes the study results and derives implications for practice, limitations, and future research ideas.
The emerging threat of antibiotic-resistant bacteria has become a global challenge in the last decades, leading to a rising demand for alternative treatments for bacterial infections. One approach is to target the bacterial cell envelope, making understanding its biophysical properties crucial. Specifically, bacteriophages use the bacterial envelope as an entry point to initiate infection, and they are considered important building blocks of new antibiotic strategies against drug-resistant bacteria.. Depending on the structure of the cell wall, bacteria are classified as Gram-negative and Gram-positive. Gram-negative bacteria are equipped with a complex cell envelope composed of two lipid membranes enclosing a rigid peptidoglycan layer. The synthesis machinery of the Gram-negative cell envelope is the target of antimicrobial agents, including new physical sanitizing procedures addressing the outer membrane (OM). It is therefore very important to study the biophysical properties of the Gram-negative bacterial cell envelope. The high complexity of the Gram-negative OM sets the demand for a model system in which the contribution of individual components can be evaluated separately. In this respect, giant unilamellar vesicles (GUVs) are promising membrane systems to study membrane properties while controlling parameters such as membrane composition and surrounding medium conditions.
The aim of this work was to develop methods and approaches for the preparation and characterization of a GUV-based membrane model that mimics the OM of the Gram-negative cell envelope. A major component of the OM is the lipopolysaccharide (LPS) on the outside of the OM heterobilayer. The vesicle model was designed to contain LPS in the outer leaflet and lipids in the inner leaflet. Furthermore, the interaction of the prepared LPS-GUVs with bacteriophages was tested. LPS containing GUVs were prepared by adapting the inverted emulsion technique to meet the challenging properties of LPS, namely their high self-aggregation rate in aqueous solutions. Notably, an additional emulsification step together with the adaption of solution conditions was employed to asymmetrically incorporate LPS containing long polysaccharide chains into the artificial membranes. GUV membrane asymmetry was verified with a fluorescence quenching assay. Since the necessary precautions for handling the quenching agent sodium dithionite are often underestimated and poorly described, important parameters were tested and identified to obtain a stable and reproducible assay. In the context of varied LPS incorporation, a microscopy-based technique was introduced to determine the LPS content on individual GUVs and to directly compare vesicle properties and LPS coverage. Diffusion coefficient measurements in the obtained GUVs showed that increasing LPS concentrations in the membranes resulted in decreased diffusivity.
Employing LPS-GUVs we could demonstrate that a Salmonella bacteriophage bound with high specificity to its LPS receptor when presented at the GUV surface, and that the number of bound bacteriophages scaled with the amount of presented LPS receptor. In addition to binding, the bacteriophages were able to eject their DNA into the vesicle lumen. LPS-GUVs thus provide a starting platform for bottom-up approaches for the generation of more complex membranes, in which the effects of individual components on the membrane properties and the interaction with antimicrobial agents such as bacteriophages could be explored.
This dissertation focuses on the understanding of the optical manipulation of microgels dispersed in aqueous solution of azobenzene containing surfactant. The work consists of three parts where each part is a systematic investigation of the (1) photo-isomerization kinetics of the surfactant in complex with the microgel polymer matrix, (2) light driven diffusiosmosis (LDDO) in microgels and (3) photo-responsivity of microgel on complexation with spiropyran.
The first part comprises three publications where the first one [P1] investigates the photo-isomerization kinetics and corresponding isomer composition at a photo-stationary state of the photo-sensitive surfactant conjugated with charged polymers or micro sized polymer networks to understand the structural response of such photo-sensitive complexes. We report that the photo-isomerization of the azobenzene-containing cationic surfactant is slower in a polymer complex compared to being purely dissolved in an aqueous solution. The surfactant aggregates near the polyelectrolyte chains at concentrations much lower than the bulk critical micelle concentration. This, along with the inhibition of the photo-isomerization kinetics due to steric hindrance within the densely packed aggregates, pushes the isomer-ratio to a higher trans-isomer concentration for all irradiation wavelengths.
The second publication [P2] combines experimental results and non-adiabatic dynamic simulations for the same surfactant molecules embedded in the micelles with absorption spectroscopy measurements of micellar solutions to uncover the reasons responsible for the slowdown in photo induced trans → cis azobenzene isomerization at concentrations higher than the critical micelle concentration (CMC). The simulations reveal a decrease of isomerization quantum yields for molecules inside the micelles and observes a reduction of extinction coefficients upon micellization. These findings explain the deceleration of the trans → cis switching in micelles of the azobenzene-containing surfactants.
Finally, the third publication [P3] focusses on the kinetics of adsorption and desorption of the same surfactant within anionic microgels in the dark and under continuous irradiation. Experimental data demonstrate, that microgels can serve as a selective absorber of the trans isomers. The interaction of the isomers with the gel matrix induces a remotely controllable collapse or swelling on appropriate irradiation wavelengths. Measuring the kinetics of the microgel size response and knowing the exact isomer composition under light exposure, we calculate the adsorption rate of the trans-isomers.
The second part comprises two publications. The first publication [P4] reports on the phenomenon of light-driven diffusioosmotic (DO) long-range attractive and repulsive interactions between micro-sized objects, whose range extends several times the size of microparticles and can be adjusted to point towards or away from the particle by varying irradiation parameters such as intensity or wavelength of light. The phenomenon is fueled by the aforementioned photosensitive surfactant. The complex interaction of dynamic exchange of isomers and photo-isomerization rate yields to relative concentrations gradients of the isomers in the vicinity of micro-sized object inducing a local diffusioosmotic (DO) flow thereby making a surface act as a micropump.
The second publication [P5] exclusively aims the visualization and investigation of the DO flows generated from microgels by using small tracer particles. Similar to micro sized objects, the flow is able to push adjacent tracers over distances several times larger than microgel size. Here we report that the direction and the strength of the l-LDDO depends on the intensity, irradiation wavelength and the amount of surfactant adsorbed by the microgel. For example, the flow pattern around a microgel is directed radially outward and can be maintained quasi-indefinitely under exposure at 455 nm when the trans:cis ratio is 2:1, whereas irradiation at 365 nm, generates a radially transient flow pattern, which inverts at lower intensities.
Lastly, the third part consists of one publication [P6] which, unlike the previous works, reports on the study of the kinetics of photo- and thermo-switching of a new surfactant namely, spiropyran, upon exposure with light of different wavelengths and its interaction with p(NIPAM-AA) microgels. The surfactant being an amphiphile, switches between its ring closed spiropyran (SP) form and ring open merocyanine (MC) form which results in a change in the hydrophilic–hydrophobic balance of the surfactant as MC being a zwitterionic form along with the charged head group, generates three charges on the molecule. Therefore, the MC form of the surfactant is more hydrophilic than in the case of the neutral SP state. Here, we investigate the initial shrinkage of the gel particles via charge compensation on first exposure to SP molecules which results from the complex formation of the molecules with the gel matrix, triggering them to become photo responsive. The size and VPTT of the microgels during irradiation is shown to be a combination of heating up of the solution during light absorption by the surfactant (more pronounced in the case of UV irradiation) and the change in the hydrophobicity of the surfactant.
Biofilms are heterogeneous structures made of microorganisms embedded in a self-secreted extracellular matrix. Recently, biofilms have been studied as sustainable living materials with a focus on the tuning of their mechanical properties. One way of doing so is to use metal ions. In particular biofilms have been shown to stiffen in presence of some metal cations and to soften in presence of others. However, the specificity and the determinants of those interactions vary between species. While Escherichia coli is a widely studied model organism, little is known concerning the response of its biofilms to metal ions. In this work, we aimed at tuning the mechanics of E. coli biofilms by acting on the interplay between matrix composition and metal cations. To do so, we worked with E. coli strains producing a matrix composed of curli amyloid fibres or phosphoethanolamine-cellulose (pEtN-cellulose) fibres or both. The viscoelastic behaviour of the resulting biofilms was investigated with rheology after incubation with one of the following metal ion solutions: FeCl3, AlCl3, ZnCl2 and CaCl2 or ultrapure water. We observed that the strain producing both fibres stiffen by a factor of two when exposed to the trivalent metal cations Al(III) and Fe(III) while no such response is observed for the bivalent cations Zn(II) and Ca(II). Strains producing only one matrix component did not show any stiffening in response to either cation, but even a small softening. In order to investigate further the contribution of each matrix component to the mechanical properties, we introduced additional bacterial strains producing curli fibres in combination with non-modified cellulose, non-modified cellulose only or neither component. We measured biofilms produced by those different strains with rheology and without any solution. Since rheology does not preserve the architecture of the matrix, we compared those results to the mechanical properties of biofilms probed with the non-destructive microindentation. The microindentation results showed that biofilm stiffness is mainly determined by the presence of curli amyloid fibres in the matrix. However, this clear distinction between biofilm matrices containing or not containing curli is absent from the rheology results, i.e. following partial destruction of the matrix architecture. In addition, rheology also indicated a negative impact of curli on biofilm yield stress and flow stress. This suggests that curli fibres are more brittle and therefore more affected by the mechanical treatments. Finally, to examine the molecular interactions between the biofilms and the metal cations, we used Attenuated total reflectance - Fourier transform infrared spectroscopy (ATR-FTIR) to study the three E.coli strains producing a matrix composed of curli amyloid fibres, pEtN-cellulose fibres or both. We measured biofilms produced by those strains in presence of each of the aforementioned metal cation solutions or ultrapure water. We showed that the three strains cannot be distinguished based on their FTIR spectra and that metal cations seem to have a non-specific effect on bacterial membranes in absence of pEtN-cellulose. We subsequently conducted similar experiments on purified curli or pEtN-cellulose fibres. The spectra of the pEtN-cellulose fibres revealed a non-valence-specific interaction between metal cations and the phosphate of the pEtN-modification. Altogether, these results demonstrate that the mechanical properties of E. coli biofilms can be tuned via incubation with metal ions. While the mechanism involving curli fibres remains to be determined, metal cations seem to adsorb onto pEtN-cellulose and this is not valence-specific. This work also underlines the importance of matrix architecture to biofilm mechanics and emphasises the specificity of each matrix composition.
My thesis chiefly aims to shed light on the favourable properties of LHP semiconductors from the point of view of their electronic structure.
Currently, various hypotheses are circulating to explain the exceptionally favourable transport properties of LHPs. Seeking an explanation for the low non-radiative recombination rates and long carrier lifetimes is particularly interesting to the halide perovskites research community.
The first part of this work investigates the two main hypotheses that are believed to play a significant role: the existence of a giant Rashba effect and large polarons. The experimental method of ARPES is mainly applied to verify their credibility.
The first hypothesis presumes that a giant Rashba effect restricts the recombination losses of the charge carriers by making the band gap slightly indirect. The Rashba effect is based on a strong SOC that could appear in LHPs thanks to incorporating the heavy element Pb in their structure. Earlier experimental work had pointed out this effect at the VBM of a hybrid LHP as a viable explanation for the long lifetimes of the charge carriers.
My systematic ARPES studies on hybrid MAPbBr3 and spin-resolved ARPES studies on the inorganic CsPbBr3 disprove the presence of any Rashba effect in the VBM of the reported order of magnitude. Therefore, neither the spin texture nor an indirect band gap character at the VBM in the bulk or at the surface can explain the high efficiency of LHP. In case of existence, this effect is in terms of the Rashba parameter at least a factor of a hundred smaller than previously assumed.
The second hypothesis proposes large polaron formation in the electronic structure of LHPs and attributes it to their high defect tolerance and low non-radiative recombination rate. Because the perovskite structure consists of negative and positive ions, polarons of this kind can be expected due to the Coulomb interaction between carriers and the polar lattice at intermediate electron-phonon coupling strength. Their existence is proposed to screen the carriers and defects to avoid recombination and trapping, thus leading to long carrier lifetimes. ARPES results by one group supported this assumption, reporting a 50% effective mass enhancement over the theoretical effective mass for CsPbBr3 in the orthorhombic structure.
The current thesis examines this hypothesis experimentally by photon-energy-dependent ARPES spectra and theoretically by GW band calculations of CsPbBr3 perovskites. The investigation is based on the fact that a polaron contribution in charge transport can become evident by an increase of the effective mass as measured by ARPES over the calculated one without polaron effects. However, my experiments on crystalline CsPbBr3 did not imply a larger effective mass for which one could postulate large polarons. In fact, the effective masses determined from ARPES agree with that of theoretical predictions.
The second part of my thesis thoroughly investigates the possibility of spontaneously magnetizing LHPs by introducing Mn2+ ions. Mn doping was reported to cause ferromagnetism in one of the most common LHPs, MAPbI3, mediated by super-exchange. The current work investigates the magnetic properties of a wide concentration range of Mn-doped MAPbI3 and triple-cation films by XAS, XMCD, and SQUID measurements. Based on the XAS line shape and a sum-rule analysis of the XMCD spectra, a pure Mn2+ configuration has been confirmed. Negative Curie temperatures are extracted from fitting the magnetization with a Curie-Weiss law. However, a remanent magnetization, which would be an indication of the absence of ferromagnetism down to 2K. As far as the double exchange is concerned, the element-specific XAS excludes a sufficient amount of Mn3+ as a prerequisite for this mechanism. All the findings show no evidence of significant double exchange or ferromagnetism in Mn-doped LHPs. The magnetic behavior is paramagnetic rather than ferromagnetic.
In the dissertation's last chapter, orthorhombic features of CsPbBr3 are revealed by ARPES, including an extra VBM at the Γ-point. The VBM of CsPbBr3 shows a temperature-dependent splitting, which decreases by 190 meV between 38K and 300K and tracks a shift of a saddle point at the cubic M-point. It is possible to reproduce the energy shift using an atomic model with a larger unit cell for room temperature, allowing local inversion symmetry breaking. This indicates the importance of electric dipoles for the inorganic LHPs, which may contribute to their high efficiency by breaking inversion symmetry and a Berry-phase effect.
The light reactions of photosynthesis are carried out by a series of multiprotein complexes embedded in thylakoid membranes. Among them, photosystem I (PSI), acting as plastocyanin-ferderoxin oxidoreductase, catalyzes the final reaction. Together with light-harvesting antenna I, PSI forms a high-molecular-weight supercomplex of ~600 kDa, consisting of eighteen subunits and nearly two hundred co-factors. Assembly of the various components into a functional thylakoid membrane complex requires precise coordination, which is provided by the assembly machinery. Although this includes a small number of proteins (PSI assembly factors) that have been shown to play a role in the formation of PSI, the process as a whole, as well as the intricacy of its members, remains largely unexplored.
In the present work, two approaches were used to find candidate PSI assembly factors. First, EnsembleNet was used to select proteins thought to be functionally related to known PSI assembly factors in Arabidopsis thaliana (approach I), and second, co-immunoprecipitation (Co-IP) of tagged PSI assembly factors in Nicotiana tabacum was performed (approach II).
Here, the novel PSI assembly factors designated CO-EXPRESSED WITH PSI ASSEMBLY 1 (CEPA1) and Ycf4-INTERACTING PROTEIN 1 (Y4IP1) were identified. A. thaliana null mutants for CEPA1 and Y4IP1 showed a growth phenotype and pale leaves compared with the wild type. Biophysical experiments using pulse amplitude modulation (PAM) revealed insufficient electron transport on the PSII acceptor side. Biochemical analyses revealed that both CEPA1 and Y4IP1 are specifically involved in PSI accumulation in A. thaliana at the post-translational level but are not essential. Consistent with their roles as factors in the assembly of a thylakoid membrane protein complex, the two proteins localize to thylakoid membranes. Remarkably, cepa1 y4ip1 double mutants exhibited lethal phenotypes in early developmental stages under photoautotrophic growth. Finally, co-IP and native gel experiments supported a possible role for CEPA1 and Y4IP1 in mediating PSI assembly in conjunction with other PSI assembly factors (e.g., PPD1- and PSA3-CEPA1 and Ycf4-Y4IP1). The fact that CEPA1 and Y4IP1 are found exclusively in green algae and higher plants suggests eukaryote-specific functions. Although the specific mechanisms need further investigation, CEPA1 and Y4IP1 are two novel assembly factors that contribute to PSI formation.
Biostimulant SuperFifty based molecular priming to increase plant strength and stress tolerance
(2023)
Depressive disorders are associated with reduced life satisfaction and ability to work. The waiting time for psychotherapy in Germany is currently between three and six months. Accordingly, there is a need for alternative, evidence-based treatment options that are made accessible to patients at a low threshold. A large number of empirical studies prove the effectiveness of exercise in mild and moderate depression. For further conceptualization and quality assurance of exercise as a treatment option, it is necessary to understand the concrete mechanisms of action. In addition to physiological factors, psychological factors also play a role in the effect. As a meta-theory of human experience and behavior, Self-Determination Theory (SDT) provides a useful frame for understanding psychological mechanisms of action with concrete implications for treatment practice. The conceptual extension of SDT to include the frustration of basic psychological needs in addition to need satisfaction is proving useful in the study of mental illness. The first part of this dissertation consists of two publications that validate relevant measurement instruments in this context. The first questionnaire measures the extent of generally experienced satisfaction and frustration of the basic psychological needs for autonomy, competence, and relatedness. The second questionnaire measures the experienced satisfaction of needs by the instructor (here: exercise therapist). The second part of the dissertation includes two publications that examine and classify the satisfaction and frustration of basic psychological needs in depressive symptoms. Differences in the extent of need satisfaction and need frustration between a sample with depression and a sample without depressive symptoms are examined. Further, the relationship between need frustration and depressive symptoms is placed in the context of established pathological processes (emotional dysregulation, rumination). The main findings of this work show that by adding the dimension of need frustration to Basic Psychological Needs Theory, SDT now covers a broader spectrum on the health-disease continuum. In doing so, SDT focuses on the psychological impact of social environments. In addition to the nonfulfillment of basic psychological needs, it is primarily experienced need frustration that is a general vulnerability factor for the occurrence of psychological illness. Moreover, the unbalanced satisfaction of basic psychological needs possibly indicates a conflicting experience between the needs. For the treatment practice it can be deduced that an autonomy-supporting atmosphere, which enables the balanced satisfaction of all three needs, is central for the treatment success.
The development of seeds in angiosperms starts with a complex process of double fertilization, involving the fusion of the maternal egg cell and central cell with two paternal sperm cells. This gives rise to the embryo and the nourishing endosperm, which are then enclosed by the seed coat, derived from the maternal integuments. The growth of the seed coat in Arabidopsis thaliana (Arabidopsis) is actively inhibited before fertilization by epigenetic regulators known as Polycomb Group (PcG) proteins. These proteins deposit a repressive histone mark called H3K27me3, which must be removed to enable seed coat formation. In this thesis, I explored the mechanism of removal of H3K27me3 marks from the integument cells following fertilization, which allows for seed coat formation. We hypothesized that this removal should be primarily facilitated by histone demethylases from the JMJ family and potentially influenced by the plant hormones Brassinosteroids (BRs). This hypothesis was supported by the expression patterns of the JMJ protein REF6 and of BR related genes, which are specifically expressed in the integuments and in the seed coat. Moreover, mutations in both these pathways lead to developmental defects, such as reduced ovule viability and delayed seed coat growth. Our research provides evidence suggesting that BR signalling is likely involved in recruiting JMJ-type histone demethylases to target loci responsible for seed coat growth. Moreover, we have discovered an additional pathway through which BRs regulate seed coat development, independent of their influence on H3K27me3 marks. This finding emphasizes the diverse roles of BRs in coordinating seed development, extending beyond their well-known involvement in plant growth and development. Furthermore, I explored the role of another epigenetic mark, DNA methylation, in fertilization-independent (or autonomous) seed formation in Arabidopsis. For this, we utilized epigenetic Recombinant Inbred Lines (epiRILs) and thus identified an epigenetic Quantitative Trait Locus (epiQTL) on chromosome II, potentially responsible for the larger autonomous seed size observed in DNA methylation mutants. Overall, this thesis significantly enhances our comprehension of the intricate relationship between epigenetic modifications, hormonal signaling, and plant reproductive processes. It offers valuable insights into the genetic mechanisms governing both sexual and asexual seed formation, while also presenting potential avenues for the engineer of advantageous traits in agricultural crops.
Interested in the ideological workings of fiction, I study how major avant-garde tropes promote the potential of permanent renewal as white America’s property. Renewal ties to the capacities to create, progress, transcend, and simply be. From Black critique we know that, within dominant discourse, all these capacities have been denied to Black bodies ever since colonization. Black work has been fetishized, appropriated, stolen, and dismissed in and by dominant culture, while Black being is construed as negativity and barred on the level of ontology. It follows then that racialization operates on multiple levels in the conceptual frame of renewal. I study this conceptualization by re-reading the works of and criticism on progressive white authors. I examine how images of renewal enable the claim on futurity, transformative potential, and movement forward as exclusively white properties. Premised on oppositions between positive capacities and a state of complete incapacitation, these images are often viewed as separate constructions. This project shows that, deriving from white ideology, such representations are symbiotic and simultaneous - the "good" story of white renewal rests on the continual transgression towards Black being.
Nowadays, innovative and entrepreneurial activities and their actors are embedded in interdependent systems to drive joint value creation. Innovation ecosystems and entrepreneurial ecosystems have become established system-level concepts in management research to explain how value transpires between different actors and institutions in distinct contexts. Despite the popularity of the concepts, researchers have critiqued their theoretical depth, conceptual distinctiveness, as well as operationalization and measurement (Autio & Thomas, 2022; Klimas & Czakon, 2022). Furthermore, in light of current-day challenges, research has yet to address how context impacts innovation and entrepreneurial ecosystems and their actors and elements (Wurth et al., 2022).
The aim of this cumulative thesis is to provide a deeper understanding of the conceptualization, operationalization, and measurement of innovation and entrepreneurial ecosystems and investigate how contextual factors can influence the overall ecosystem and its key actors. To this end, bibliometric and empirical-qualitative methods, as well as narrative and systematic literature reviews, are employed. After introducing the research scope and key concepts in Chapter 1, a systematic literature review to operationalize and measure the concept of innovation ecosystems is conducted, and an integrative framework of its composition is introduced in Chapter 2. In Chapter 3, the innovation journal network is outlined by means of science mapping to determine current and emerging research areas characterizing innovation studies. In Chapters 4 and 5, the interplay between the temporal context of the Covid-19 pandemic and the spatial context of entrepreneurial ecosystems is assessed by focusing on the role of organizational resilience and affordances. The findings shed new light on the dynamics and boundaries of entrepreneurial ecosystems as they move between the spatial and digital realm. Building on this, an integrative framework of digital entrepreneurial ecosystems is presented in Chapter 6. The concluding Chapter 7 summarizes my thesis’s conceptual, theoretical, and empirical insights, highlighting implications, limitations, and promising future research avenues.
The findings of this cumulative thesis contribute to the theoretical and conceptual advancement of ecosystems in innovation and entrepreneurship by providing insights into the measurement and operationalization of its elements. Furthermore, the results show that contextual factors, such as crisis events or institutional circumstances, influence innovation and entrepreneurial ecosystems and their actors, calling for a more nuanced consideration of ecosystem configurations and dynamics. By drawing from the theory of affordances, the elements that actually afford value to the actors and how they shift between the physical and digital realm are portrayed. Based on these findings, this thesis introduces novel frameworks and conceptual advancements of the configurations and boundaries of innovation and (digital) entrepreneurial ecosystems, laying the foundation for a renewed understanding of how to design, orchestrate, and evaluate ecosystems today and in the future.
Aging is a complex process characterized by several factors, including loss of genetic and epigenetic information, accumulation of chronic oxidative stress, protein damage and aggregates and it is becoming an emergent drug target. Therefore, it is the utmost importance to study aging and agerelated diseases, to provide treatments to develop a healthy aging process. Skeletal muscle is one of the earliest tissues affected by age-related changes with progressive loss of muscle mass and function from 30 years old, effect known as sarcopenia. Several studies have shown the accumulation of protein aggregates in different animal models, as well as in humans, suggesting impaired proteostasis, a hallmark of aging, especially regarding degradation systems. Thus, different publications have explored the role of the main proteolytic systems in skeletal muscle from rodents and humans, like ubiquitin proteasomal system (UPS) and autophagy lysosomal system (ALS), however with contradictory results. Yet, most of the published studies are performed in muscles that comprise more than one fiber type, that means, muscles composed by slow and fast fibers. These fiber types, exhibit different metabolism and contraction speed; the slow fibers or type I display an oxidative metabolism, while fast fibers function towards a glycolytic metabolism ranging from fast oxidative to fast glycolytic fibers. To this extent, the aim of this thesis sought to understand on how aging impacts both fiber types not only regarding proteostasis but also at a metabolome and transcriptome network levels. Therefore, the first part of this thesis, presents the differences between slow oxidative (from Soleus muscle) and fast glycolytic fibers (Extensor digitorum longus, EDL) in terms of degradation systems and how they cope with oxidative stress during aging, while the second part explores the differences between young and old EDL muscle transcriptome and metabolome, unraveling molecular features. More specifically, the results from the present work show that slow oxidative muscle performs better at maintaining the function of UPS and ALS during aging than EDL muscle, which is clearly affected, accounting for the decline in the catalytic activity rates and accumulation of autophagy-related proteins. Strinkingly, transcriptome and metabolome analyses reveal that fast glycolytic muscle evidences significant downregulation of mitochondrial related processes and damaged mitochondria morphology during aging, despite of having a lower oxidative metabolism compared to oxidative fibers. Moreover, predictive analyses reveal a negative association between aged EDL gene signature and lifespan extending interventions such as caloric restriction (CR). Although, CR intervention does not alter the levels of mitochondrial markers in aged EDL muscle, it can reverse the higher mRNA levels of muscle damage markers. Together, the results from this thesis give new insights about how different metabolic muscle fibers cope with age-related changes and why fast glycolytic fibers are more susceptible to aging than slow oxidative fibers.
The increasing global population has led to a growing demand for cost-effective and eco-friendly methods of water purification. This demand has reached a peak due to the increasing presence of impurities and pollutants in water and a growing awareness of waterborne diseases. Advanced oxidation processes (AOPs) are effective methods to address these challenges, due to the generation of highly reactive radicals, such as sulfate radical (SO4•-), hydroxyl radical (•OH), and/or superoxide radical (•O2-) in oxidation reactions. Relative to conventional hydrogen peroxide (H2O2)-based AOPs for wastewater treatment, the persulfate-related AOPs are receiving increasing attention over the past decades, due to their stronger oxidizing capability and a wider pH working window. Further deployment of the seemingly plausible technology as an alternative for the well-established one in industry, however, necessitates a careful evaluation of compounding factors, such as water matrix effects, toxicological consequences, costs, and engineering challenges, etc. To this end, rational design of efficient and environmentally friendly catalysts constitutes an indispensable pathway to advance persulfate activation efficacy and to elucidate the mechanisms in AOPs, the combined endeavors are expected to provide insightful understanding and guidelines for future studies in wastewater treatment. A dozens of transition metal-based catalysts have been developed for persulfate-related AOPs, while the undesirable metal leaching and poor stability in acidic conditions have been identified as major obstacles. Comparatively, the carbonaceous materials are emerging as alternative candidates, which are characterized by metal-free nature, wide availability, and exceptional resistance to acid and alkali, as well as tunable physicochemical and electronic properties, the combined merits make them an attractive option to overcome the aforementioned limitations in metal-based catalytic systems. This dissertation aims at developing novel carbonaceous materials to boost the activity in peroxymonosulfate (PMS) activation processes. Functionalized carbon materials with metal particles or heteroatoms were constructed and further evaluated in terms of their ability to activate PMS for AOPs. The main contents of this thesis are summarized as follows: (1) Iron oxide-loaded biochar: improving stability and alleviating metal leakage Metal leaching constitutes one of the main drawbacks in using transition metals as PMS activators, which is accompanied by the generation of metal-containing sludge, potentially leading to secondary pollution. Meanwhile, the metal nanoparticles are prone to aggregate, causing rapid decay of catalytic performance. The use of carbons as supports for transition metals could mitigate these deficiencies, because the interaction between metals and carbons could in turn disperse and stabilize metal nanoparticles, thus suppressing the metal leaching. In this work, the environmentally benign lignin with its abundant phenolic groups, which is well known to serve as carbon source with high yields and flexibility, was utilized to load Fe ions. The facile low-temperature pre-treatment pyrolytic strategy was employed to construct a green catalyst with iron oxides embedded in Kraft-lignin-derived biochar (termed as γ-Fe2O3@KC). The γ-Fe2O3@KC was capable of activating PMS to generate stable non-radical species (1O2 and Fe (V)=O) and to enhance electron transfer efficiency. A surface-bound reactive complex (catalyst-PMS*) was identified by electrochemical characterizations and discussed with primary surface-bound radical pairs to explain the contradictions between quenching and EPR detection results. The system also showed encouraging reusability for at least 5 times and high stability at pH 3-9. The low concentration of iron in γ-Fe2O3@KC/PMS system implied that the carbon scaffold of biochar substantially alleviated metal leakage. (2) MOF-derived nanocarbon: new carbon crystals Traditional carbon materials are of rather moderate performance in activation PMS, due to the poor electron transfer capacity within the amorphous structure and limited active sites for PMS adsorption. Herein, we established crystalline nanocarbon materials via a simple NaCl-templated strategy using the monoclinic zeolitic imidazolate framework-8 (ZIF-8) sealed with NaCl crystals as the precursors. Specifically, NaCl captured dual advantages in serving as structure-directing agent during hydrolysis and protective salt reactor to facilitate phase transformation during carbonization. The structure-directing agent NaCl provided a protective and confined space for the evolution of MOF upon carbonization, which led to high doping amounts of nitrogen (N) and oxygen elements (O) in carbon framework (N: 14.16 wt%, O: 9.6 wt%) after calcination at a high temperature of 950 oC. We found that N-O co-doping can activate the chemically inert carbon network and the nearby sp2-hybridized carbon atoms served as active sites for adsorption and activation. Besides, the highly crystallized structure with well-established carbon channels around activated carbon atoms could significantly accelerate electron transfer process after initial adsorption of PMS. As such, this crystalline nanocarbon exhibited excellent catalytic kinetics for various pollutants. (3) MOF-derived 2D carbon layers: enhanced mass/electron transfer The two-dimensional (2D) configuration of carbon-based nanosheets with inherent nanochannels and abundant active sites residing on the layer edges or in between the layers, allowed the accessible interaction and close contact between the substrates and reactants, as well as the dramatically improved electron- and mass-transfer kinetics. In this regard, we developed dual-templating strategy to afford 2D assembly of the crystalline carbons, which found efficiency in reinforcing the interactions between the catalyst surface and foreign pollutants. Specifically, we found that the ice crystals and NaCl promoted the evolution of MOF in a 2D fashion during the freezing casting stage, while the later further allowed the formation of a graphitic surface at high calcination temperature, by virtue of the templating effect of molten salt. Due to the highly retained co-doping amounts, N and O heteroatoms created abundant active sites for PMS activation, the 2D configuration of carbon-based nanosheets enable efficient interaction of PMS and pollutants on the surface, which further boosted the kinetics of degradation.
The musculoskeletal system provides support and enables movement to the body, and its deterioration is a crucial aspect of age-related functional decline. Mesenchymal stromal cells (MSCs) play an important role in musculoskeletal homeostasis due to their broad differentiation potentials and their ability to support osteogenic and myogenic tissue maintenance and regeneration. In the bone, MSCs differentiate either into osteochondrogenic progenitors to form osteocytes and chondrocytes, or increasingly with age into adipogenic progenitors which give rise to bone-resident adipocytes. In skeletal muscle, during healthy regeneration MSCs provide regulatory signals that activate local, tissue-specific stem cells, known as satellite cells, which regenerate contractile myofibres. This process involves a significant cross-talk to immune cells stemming from both lymphoid and myeloid lineages. During ageing, muscle-resident MSCs undergo increased adipogenic lineage commitment, causing niche changes that contribute to fatty infiltration in muscles. These shifts in cell populations in bone lead to the loss of osteogenic cells and subsequently osteoporosis, or in muscle to impaired regeneration and to the development of sarcopenia. However, the signals that drive transition of MSCs into their respective cellular fates remain elusive.
This thesis aims to elucidate the transcriptional shifts modulating cell states and cell types in musculoskeletal MSC fate determination. Single-cell RNA-sequencing (scRNA-seq) was used to characterise cell type-specific transcript regulation. State-of-the-art bioinformatics tools were combined with different analytical platforms that include both droplet-based scRNA-seq for large heterogeneous populations, and microfluidics-based scRNA-seq to assess small, rare subpopulations. For each platform, distinct computational pipelines were established including filtering steps to exclude low-quality cells, and data visualisation was performed by dimensionality reduction. Downstream analysis included clustering, cell type annotation, and differential gene expression to investigate transcriptional states in defined cell types during ageing and injury in the muscle and bone. Finally, a novel tool to assess publication activities in defined areas of research for the identified marker genes was developed.
The results in the bone indicate that ageing MSCs increasingly commit towards an adipogenic fate at the expense of osteogenic specialisation. The data also suggests that significant cell population shifts of MSC-type fibro-adipogenic progenitors during muscle ageing underlie the pathologies observed in homeostatic and post-injury regenerative conditions. High-throughput visualisation of publication activity for candidate genes enabled more effective biological evaluation of scRNA-seq data. These results expose critical age-related changes in the stem cell niches of skeletal muscle and bone, highlight their respective sensitivity to nutrition and pathology, and elucidate novel factors that modulate stem cell-based regeneration. Targeting these processes might improve musculoskeletal health in the context of ageing and prevent the negative effects of pathological lineage determination.
In times of ongoing biodiversity loss, understanding how communities are structured and what mechanisms and local adaptations underlie the patterns we observe in nature is crucial for predicting how future ecological and anthropogenic changes might affect local and regional biodiversity. Aquatic zooplankton are a group of primary consumers that represent a critical link in the food chain, providing nutrients for the entire food web. Thus, understanding the adaptability and structure of zooplankton communities is essential. In this work, the genetic basis for the different temperature adaptations of two seasonally shifted (i.e., temperature-dependent) occurring freshwater rotifers of a formerly cryptic species complex (Brachionus calyciflorus) was investigated to understand the overall genetic diversity and evolutionary scenario for putative adaptations to different temperature regimes. Furthermore, this work aimed to clarify to what extent the different temperature adaptations may represent a niche partitioning process thus enabling co-existence. The findings were then embedded in a metacommunity context to understand how zooplankton communities assemble in a kettle hole metacommunity located in the northeastern German "Uckermark" and which underlying processes contribute to the biodiversity patterns we observe. Using a combined approach of newly generated mitochondrial resources (genomes/cds) and the analysis of a candidate gene (Heat Shock Protein 40kDa) for temperature adaptation, I showed that the global representatives of B. calyciflorus s.s.. are genetically more similar than B. fernandoi (average pairwise nucleotide diversity: 0.079 intraspecific vs. 0.257 interspecific) indicating that both species carry different standing genetic variation. In addition to differential expression in the thermotolerant B. calyciflorus s.s. and thermosensitive B. fernandoi, the HSP 40kDa also showed structural variation with eleven fixed and six positively selected sites, some of which are located in functional areas of the protein. The estimated divergence time of ~ 25-29 Myr combined with the fixed sites and a prevalence of ancestral amino acids in B. calyciflorus s.s. indicate that B. calyciflorus s.s. remained in the ancestral niche, while B. fernandoi partitioned into a new niche. The comparison of mitochondrial and nuclear markers (HPS 40kDa, ITS1, COI) revealed a hybridisation event between the two species. However, as hybridisation between the two species is rare, it can be concluded that the temporally isolated niches (i.e., seasonal-shifted occurrence) they inhabit based on their different temperature preferences most likely represent a pre-zygotic isolation mechanism that allows sympatric occurrence while maintaining species boundaries. To determine the processes underlying zooplankton community assembly, a zooplankton metacommunity comprising 24 kettle holes was sampled over a two-year period. Active (i.e., water samples) and dormant communities (i.e., dormant eggs hatched from sediment) were identified using a two-fragment DNA metabarcoding approach (COI and 18S). Species richness and diversity as well as community composition were analysed considering spatial, temporal and environmental parameters. The analysis revealed that environmental filtering based on parameters such as pH, size and location of the habitat patch (i.e., kettle hole) and surrounding field crops largely determined zooplankton community composition (explained variance: Bray-Curtis dissimilarities: 10.5%; Jaccard dissimilarities: 12.9%), indicating that adaptation to a particular habitat is a key feature of zooplankton species in this system. While the spatial configuration of the kettle holes played a minor role (explained variance: Bray-Curtis dissimilarities: 2.8% and Jaccard dissimilarities: 5.5%), the individual kettle hole sites had a significant influence on the community composition. This suggests monopolisation/priority effects (i.e., dormant communities) of certain species in individual kettle holes. As environmental filtering is the dominating process structuring zooplankton communities, this system could be significantly influenced by future land-use change, pollution and climate change.
Light is the essential energy source for plants to drive photosynthesis. In nature, light availability is highly variable and often fluctuates on very short time scales. As a result, plants developed mechanisms to cope with these fluctuations. Understanding how to improve light use efficiency in natural fluctuating light (FL) conditions is a major target for agronomy.
In the first project, we identified an Arabidopsis thaliana plant that showed reduced levels of rapidly inducible non-photochemical quenching (NPQ). This plant was devoid of any T-DNA insertion. Using a mapping-by-sequencing approach, we successfully located the causal genomic region near the end of chromosome 4. Through variant investigations in that region, we identified a deletion of about 20 kb encompassing 9 genes. By complementation analysis, we confirmed that one of the deleted genes, VTC2, is the causal gene responsible for the low NPQ. Loss of VTC2 decreased NPQ particularly in old leaves, with young leaves being only slightly affected. Additionally, ascorbate levels were almost abolished in old leaves, likely causing the NPQ decrease by reducing the activity of the xanthophyll cycle. Although ascorbate levels in younger leaves were reduced compared to wild-type plants, they remained at a comparably higher level. This difference may be due to the VTC2 paralog VTC5, which is expressed at a higher level in young leaves than in old ones.
Plants require the PROTON GRADIENT REGULATION 5 (PGR5) protein for survival in FL. pgr5 mutants die because they fail to increase the luminal proton concentration in response to high light (HL) phases. A rapid elevation in ∆pH is needed to slow down electron transport through the Cytochrome b6 f complex (photosynthetic control). In FL, such lack of control in the pgr5 mutants results in photosystem I (PSI) overreduction, reactive oxygen species (ROS) production, and cell death. Decreases in photosystem II (PSII) activity introduced by crossing pgr5 with PSII deficient mutants
rescued the lethality of pgr5 in FL. PGR5 was suggested to act as part of the ferredoxin-plastoquinone reductase (FQR), involved in cyclic electron transfer around PSI. However, the proposed molecular role of PGR5 remains highly debated. To learn more about PGR5 function, we performed a forward genetic screen in Arabidopsis thaliana to identify EMS-induced suppressor mutants surviving longer when grown in FL compared to pgr5 mutants (referred to as ”suppressor of pgr5 lethality in fluctuating light”, splf ). 11 different candidate genes were identified in a total of 22 splf plants.
Mutants of seven of these genes in the pgr5 background showed low Fv/Fm values when grown in non-fluctuating low light (LL). Five of these 4genes were previously reported to have a role in PSII biogenesis or function. Two others, RPH1 and a DEAD/DEAH box helicase (AT3G02060), have not been linked to PSII function before. Three of splf candidate genes link to primary metabolism, fructose-2,6-bisphosphatase (F2KP ), udp-glucose pyrophosphorylase 1 (UGP1 ) and ferredoxin-dependent glutamate synthase (Fd-GOGAT ). They are characterized by the fact that they survive longer in FL than pgr5 mutants but do not procede beyond the early vegetative
phase and then die.
Answer Set Programming (ASP) allows us to address knowledge-intensive search and optimization problems in a declarative way due to its integrated modeling, grounding, and solving workflow. A problem is modeled using a rule based language and then grounded and solved. Solving results in a set of stable models that correspond to solutions of the modeled problem. In this thesis, we present the design and implementation of the clingo system---perhaps, the most
widely used ASP system. It features a rich modeling language originating from the field of knowledge representation and reasoning, efficient grounding algorithms based on database evaluation techniques, and high performance solving algorithms based on Boolean satisfiability (SAT) solving technology.
The contributions of this thesis lie in the design of the modeling language, the design and implementation of the grounding algorithms, and the design and implementation of an Application Programmable Interface (API) facilitating the use of ASP in real world applications and the implementation of complex forms of reasoning beyond the traditional ASP workflow.
Heat stress (HS) is one of the major abiotic stresses which adversely affects the survival and growth of plants due to their sessile nature. To combat the detrimental effects of HS and develop thermotolerance, plants have evolved several defense mechanisms. Thermomemory is one such molecular mechanism whereby plants that have been acclimated (or primed/P) by a moderate HS can respond more efficiently and continue their growth after exposure to a severe or lethal HS (called triggering/T), while unprimed plants cannot survive. Thermomemory is known to be regulated by several transcription factors (TFs), epigenetic changes, chromatin remodellers, post-transcriptional changes and it also involves protein stability control and primary metabolism adjustment. Recent research has suggested that the shoot apical meristem (SAM) in Arabidopsis thaliana has a distinct transcriptional thermomemory which is possibly regulated by eight TFs called HEAT SHOCK FACTORS (HSFs). The main objective of this PhD thesis is to investigate the role of HSFA7b (one of the eight HSFs), in regulating thermomemory at the SAM by identifying the molecular networks it regulates. HSFA7a, a close homolog of HSFA7b, is also one of the eight HSFs that are involved in regulating thermomemory at the SAM. Thermomemory was found to be defective in the hsfa7b and hsfa7a hsfa7b mutants; the percentage survival of these seedlings was significantly lower than in wild-type (WT) seedlings after the priming and triggering (PT) treatment. Transcriptome and ChIP analyses were performed to identify the molecular networks controlled by HSFA7b and its close homolog HSFA7a, in regulating thermomemory at the SAM. The chromatin regulator SPLAYED (SYD) was found to be regulated by both HSFA7a and HSFA7b at the SAM during thermomemory. SYD is directly involved in SAM maintenance by directly regulating WUSCHEL (WUS), a master regulator of stem cell maintenance. WUS expression was down-regulated at the SAM of PT treated hsfa7a/b mutants compared to WT-Col-0 seedlings. HSFA7a and HSFA7b also jointly regulate the expression of orphan gene QUA QUINE STARCH (QQS) during thermomemory. Starch accumulation negatively correlates with QQS expression and this trend was observed in WT plants in response to thermopriming. The remobilization of starch was affected in the hsfa7a/b mutants compared to WT plants during the recovery period after T treatment. These findings indicate that defects in SAM maintenance and starch remobilization could possibly contribute to the reduced thermomemory in the hsfa7a/b mutants. Moreover, transcriptome and ChIP analysis indicate that ethylene signaling genes are directly regulated by HSFA7b during thermomemory. Transcriptome analysis of the HSFA7b-IOE line indicates that HSFA7b positively regulates the expression of HEAT STRESS ASSOCIATED 32 (HSA32), an important thermomemory gene, and HSFA7b strongly suppresses the expression of the reactive oxygen species (ROS) responsive REDOX RESPONSIVE TRANSCRIPTION FACTOR 1 (RRTF1) gene, which is also a repressed target of SYD. In Arabidopsis, the HSFA7b transcript undergoes alternative splicing at high temperatures to form two splice variants: one correctly/constitutively spliced variant which is functional and codes for the HSFA7b protein and one intron retained splice variant. Higher accumulation of the functional HSFA7b splice variant was found at the SAM compared to other tissues. Moreover, accumulation of the functional splice variant was higher in P and PT plants compared to control plants, whereas higher levels of the intron retained splice variant is found in plants subjected directly to the T treatment. The intron retained HSFA7b splice variant is degraded by the non-sense mediated decay (NMD) pathway as a means of regulating transcript level essential for protein synthesis at high temperatures. Importantly, HSFA7b protein accumulation was observed in plants subjected to PT treatment that survive and continue growth, but not in plants subjected directly to T treatment that do not survive, indicating that constitutive/ correct splicing of the HSFA7b transcript is a component of thermomemory. Taken together, these findings suggest that HSFA7a and HSFA7b jointly regulate SAM maintenance via the chromatin remodeller SYD and starch remobilization via QQS. In addition to them, HSFA7b also regulates the expression of ethylene signaling genes, heat responsive genes and the ROS responsive RRTF1. Furthermore, constitutive/correct splicing in the HSFA7b transcript is also an essential component of thermomemory.
In nature, plants often encounter biotic and abiotic stresses, which can cause reduced crop yield and quality, and threaten the nutrition of a growing human population. As heat stress (HS) is one of the main abiotic stresses, and is projected to increase due to global warming, it is necessary to better understand how plants respond and survive under HS. In Arabidopsis thaliana, plants can survive under severe HS if primed by a non-lethal HS, a process called acquisition of thermotolerance. This primed stated can be maintained for several days, and the ability of plants to maintain the primed state is called maintenance of acquired thermotolerance (mATT) or HS memory. According to current research, two Heat shock factors (HSFs) HSFA2 and HSFA3 are known to account for the majority of mATT capability, and there are other HSFs e.g. HSFA1b and HSFA6b in HSF complexes containing HSFA2 and/or HSFA3, however, the roles of these HSFs in HS memory is not clearly understood. Moreover, the mechanism of these HSFs in regulating HS memory is unclear, whether transcriptional machinery e.g. the Mediator complex contributes to transcriptional memory. This work investigates the role of HSFs and Mediator subunits in HS memory in A. thaliana. For the role of HSFs, the interaction between HSFA1b and HSFA2 during HS memory phase was confirmed by in vivo co- immunoprecipitation (Co-IP). HSFA1b, HSFA2, HSFA3 and HSFA6b targeted HS memory-related genes according to DNA affinity purification sequencing (DAP-seq) data, and targets of HSFA1b were confirmed in vivo by chromatin immunoprecipitation qPCR (ChIP-qPCR). The mutant of hsfa6b showed an HS memory deficiency phenotype in mATT survival assay. These data confirmed the role for HSFA2 and HSFA3 in HS memory, and suggest that HSFA1b and HSFA6b also function in HS memory. The Mediator complex functions as an RNA Polymerase II (RNA Pol II) co-regulator, and includes Head, Middle, Tail and Kinase modules. Both MED23 and MED32 belong to the Tail module, and they have a positive role in HS memory. MED23 interacted with HSFA3, as determined by yeast two hybrid (Y2H) and in vivo Co-IP assays. The med23 mutant showed a decreased HS memory phenotype, reduced expression of Type I (sustained expression) memory genes following HS, and reduced accumulation of the memory-associated Tri-methylation of histone H3 lysine 4 (H3K4me3)histone modification at HS memory-related gene loci after HS. MED23 was recruited to HS-inducible memory and non-memory genes after HS, as determined by ChIP-qPCR. The med32
mutant showed a reduced HS memory phenotype, decreased expression of Type I and Type II (hyper-induction) memory genes, and lower accumulation of H3K4me3 at memory gene lociafter HS. However, MED32 did not show interaction with any tested HSF in Y2H or in vivo Co-IP. MED32 regulated the recruitment of RNA Pol II at HS-inducible genes after HS, but was not itself recruited to HS memory genes after HS. These results provided more evidence
that the Mediator subunits MED23 and MED32 regulate HS memory on transcriptional and epigenetic levels. In general, this work provides a better insight into the molecular mechanism of how HSFs and Mediator subunits regulate HS memory in plants and will provide new perspectives to breed crops with improved thermotolerance.
Teacher self-efficacy is highly relevant for effective teaching, student academic development, and teachers’ wellbeing, as theoretical work (Bandura, 1997; Tschannen-Moran et al., 1998) and empirical studies (i.e., Burić & Kim, 2020; Klassen & Chiu, 2010; van Uden et al., 2013) have shown associations with effective classroom management, student support, student motivation, and teachers’ job satisfaction. Given the importance of teacher self-efficacy for teaching and learning, it is interesting to note that existing research is limited in several aspects – first, longitudinal studies on the relations between teacher self-efficacy and student academic outcomes are widely missing; second, empirical studies often assess teacher self-efficacy as a one-dimensional construct, neglecting its multidimensional character; third, studies often only focus on either the students’ or the teachers’ perspective on teaching and thus do not include simultaneously multiple sources of information on teaching practices as a consequence of teacher self-efficacy; fourth, research often does not systematically consider the effects of teacher self-efficacy on students’ academic outcomes at both the group and individual level. Against this backdrop, this dissertation presents three longitudinal studies that aim to contribute to a more detailed perspective on teacher self-efficacy by examining systematically and longitudinally the relations between multiple dimensions of teacher self-efficacy and theoretically aligned teaching quality facets as perceived by teachers and students, as well as their relations to students’ motivational-affective outcomes – more concretely, their relations to students’ interest, enjoyment, and self-efficacy in a multi-level analysis approach. Study 1 examines the longitudinal relations between teacher self-efficacy for classroom management and student-perceived monitoring and social relatedness, and investigates whether the two student-reported teaching dimensions explain the relations between teacher self-efficacy for classroom management and student enjoyment using multi-level analyses. Study 2 examines longitudinally how teacher self-efficacy for student engagement relates to student interest through student- and teacher-perceived emotional support. Also, including student and teacher perspectives on teaching, Study 3 examines the longitudinal relations between specific facets of teacher self-efficacy for classroom management and classroom discipline, teacher self-efficacy for instructional strategies and cognitive activation, teacher self-efficacy for student engagement and competence support, teacher self-efficacy for emotional support and social relatedness, as well as teacher self-efficacy for student-oriented teaching and learning goal-oriented instruction with students’ enjoyment and self-efficacy. Furthermore, in Studies 2 and 3 the cross-level mediation effects of longitudinal relations with teacher self-efficacy on student motivational-affective characteristics via teaching practices are examined in order to address the lack of empirical studies disentangling the relations for specifically individual- vs. group-level effects. The limitations and implications are discussed in terms of their theoretical and practical relevance for school practice, teacher education, and teacher training.
A biological trade-off situation denotes the dependence between traits whereby an increase in the value of one of the traits leads to a decrease in the value of at least one of the others. Understanding trade-offs in cellular systems is relevant to understanding the limits and constraints to tuning desired phenotypes. Therefore, it is mainly the case for rates (i.e. fluxes) of biochemical reactions that shape not only molecular traits, like metabolite concentrations but also determine physiological traits, like growth. Intracellular fluxes are the final phenotype from transcriptional and (post)translational regulation. Quantifying intracellular fluxes provides insights into cellular physiology under particular growth conditions and can be used to characterize the metabolic activity of different pathways. However, estimating fluxes from labelling experiments is labour-intensive; therefore, developing approaches to accurately and precisely predict intracellular fluxes is essential. This thesis addresses two main problems: (i) identifying flux trade-offs and (ii) predicting accurate and precise reaction flux at a genome-scale level. To this end, the concept of an absolute flux trade-off is defined, and a constraint-based approach, termed FluTO, was developed to identify absolute flux trade-offs. FluTO is cast as a mixed integer programming approach applied to genome-scale metabolic models of E. coli, S. cerevisiae, and A. thaliana, imposing realistic constraints on growth and nutrient uptake.. The findings showed that trade-offs are not only species-specific but also specific to carbon sources. In addition, we found that different models of a single species have a different number of reactions in trade-offs. We also showed that absolute flux trade-offs depend on the biomass reaction used to model the growth of A. thaliana under different carbon and nitrogen conditions. Findings reflect the strong relation between nitrogen, carbon, and sulphur metabolisms in the leaves of C3 plants. The concept of relative trade-offs was introduced to further study trade-offs in metabolic networks. A constraint-based approach, FluTOr, was proposed to identify reactions whose fluxes are in relative trade-off concerning an optimized fitness-related cellular task, like growth. FluTOr was employed to find the relative flux trade-offsin the genome-scale metabolic networks of E. coli, S. cerevisiae, and A. thaliana. The results showed that in contrast to the A. thaliana model, the relative trade-offs in the two microorganisms depend on the carbon source, reflecting the differences in the underlying metabolic network. Furthermore, applying FluTOr also showed that reactions that participated in relative trade-offs were implicated in cofactor biosynthesis in the two microorganisms. Prediction of reaction fluxes in the constraint-based metabolic framework is usually performed by parsimonious flux balance analysis (pFBA), employing the principle of efficient usage of protein resources. However, we argued that principles related to the coordination of flux values, neglected in previous studies, provide other means to predict intracellular fluxes. To this end, we designed a constraint-based approach, termed complex-balanced FBA (cbFBA), to predict steady-state flux distributions that maximize the number of balanced complexes in a flux distribution, whereby multi-reaction dependencies are maximized. The comparative analysis showed a better agreement of the flux distributions resulting from cbFBA compared to pFBA with experimentally measured fluxes from 17 E. coli strains and 26 S. cerevisiae knock-out mutants. The results also showed that the predictions from cbFBA are more precise than those from pFBA since cbFBA results in a smaller space of alternative solutions than pFBA.
Pichia pastoris (syn. Komagataella phaffi) is a distinguished expression system widely used in industrial production processes. Recent molecular research has focused on numerous approaches to increase recombinant protein yield in P. pastoris. For example, the design of expression vectors and synthetic genetic elements, gene copy number optimization, or co-expression of helper proteins
(transcription factors, chaperones, etc.). However, high clonal variability of transformants and low screening throughput have hampered significant success.
To enhance screening capacities, display-based methodologies inherit the potential for efficient isolation of producer clones via fluorescence-activated cell sorting (FACS). Therefore, this study focused on developing a novel clone selection method that is based on the non-covalent attachment of Fab fragments on the P. pastoris cell surface to be applicable for FACS.
Initially, a P. pastoris display system was developed, which is a prerequisite for the surface capture of secreted Fabs. A Design of Experiments approach was applied to analyze the influence of various genetic elements on antibody fragment display. The combined P. pastoris formaldehyde dehydrogenase promoter (PFLD1), Saccharomyces cerevisiae invertase 2 signal peptide (ScSUC2), - agglutinin (ScSAG1) anchor protein, and the ARS of Kluyveromyces lactis (panARS) conferred highest display levels.
Subsequently, eight single-chain variable fragments (scFv) specific for the constant part of the Fab heavy or light chain were individually displayed in P. pastoris. Among the tested scFvs, the anti-human CH1 IgG domain scFv allowed the most efficient Fab capture detected by flow cytometry.
Irrespective of the Fab sequence, exogenously added as well as simultaneously secreted Fabs were successfully captured on the cell surface. Furthermore, Fab secretion capacities were shown to correlate to the level of surface-bound Fabs as demonstrated for characterized producer clones.
Flow-sorted clones presenting high amounts of Fabs showed an increase in median Fab titers (factor of 21 to 49) compared to unsorted clones when screened in deep-well plates. For selected candidates, improved functional Fab yields of sorted cells vs. unsorted cells were confirmed in an upscaled shake flask production. Since the scFv capture matrix was encoded on an episomal plasmid with inherently unstable autonomously replicating sequences (ARS), efficient plasmid curing was observed after removing the selective pressure. Hence, sorted clones could be immediately used for production without the need to modify the expression host or vector. The resulting switchable display/secretion system provides a streamlined approach for the isolation of Fab producers and subsequent Fab production.
Potato FLC-like and SVP-like proteins jointly control growth and distinct developmental processes
(2023)
Based on worldwide consumption, Solanum tuberosum L. (potato) is the most important non-grain food crop. Potato has two ways of stable propagation: sexually via flowering and vegetatively via tuberization. Remarkably, these two developmental processes are controlled by similar molecular regulators and mechanisms. Given that FLC and SVP genes act as key flowering regulators in the model species Arabidopsis and in various other crop species, this study aimed at identifying FLC and SVP homologs in potato and investigating their roles in the regulation of plant development, with a particular focus on flowering and tuberization. Our analysis demonstrated that there are five FLC-like and three SVP like proteins encoded in the potato genome. The expression profiles of StFLCs and StSVPs throughout potato development and the detected interactions between their proteins indicate tissue specificity of the individual genes and distinct roles of a variety of putative protein complexes. In particular, we discovered that StFLC-D, as well as StFLC-B, StSVP-A, and StSVP-B play a complex role in the regulation of flowering time, as not only increased but also decreased levels of their transcripts promote earlier flowering. Most importantly, StFLC-D has a marked impact on tuberization under non-inductive conditions and susceptibility to temperature-induced tuber malformation, also known as second growth. Plants with decreased levels of StFLC-D demonstrated a strong ability to produce tubers under long days and appeared to be insensitive to temperature-induced second growth. Lastly, our data also suggests that StFLCs and StSVPs may be involved in the nitrogen-dependent regulation of potato development. Taken together, this study highlights the functional importance of StFLC and StSVP genes in the regulation of distinct developmental processes in potato.
Mitochondria and plastids are organelles with an endosymbiotic origin. During evolution, many genes are lost from the organellar genomes and get integrated in the nuclear genome, in what is known as intracellular/endosymbiotic gene transfer (IGT/EGT). IGT has been reproduced experimentally in Nicotiana tabacum at a gene transfer rate (GTR) of 1 event in 5 million cells, but, despite its centrality to eukaryotic evolution, there are no genetic factors known to influence the frequency of IGT in higher eukaryotes. The focus of this work was to determine the role of different DNA repair pathways of double strand break repair (DSBR) in the integration step of organellar DNA in the nuclear genome during IGT. Here, a CRISPR/Cas9 mutagenesis strategy was implemented in N. tabacum, with the aim of generating mutants in nuclear genes without expected visible phenotypes. This strategy led to the generation of a collection of independent mutants in the LIG4 (necessary for non-homologous end joining, NHEJ) and POLQ genes (necessary for microhomology mediated end joining, MMEJ). Targeting of other DSBR genes (KU70, KU80, RPA1C) generated mutants with unexpectedly strong developmental phenotypes.. These factors have telomeric roles, hinting towards a possible relationship between telomere length, and strength of developmental disruption upon loss of telomere structure in plants. The mutants were made in a genetic background encoding a plastid-encoded IGT reporter, that confers kanamycin resistance upon transfer to the nucleus. Through large scale independent experiments, increased IGT from the chloroplast to the nucleus was observed in lig4 mutants, as well as lines encoding a POLQ gene with a defective polymerase domain (polqΔPol). This shows that NHEJ or MMEJ have a double-sided relationship with IGT: while transferred genes may integrate using either pathway, the presence of both pathways suppresses IGT in wild-type somatic cells, thus demonstrating for the first time the extent on which nuclear genes control IGT frequency in plants. The IGT frequency increases in the mutants are likely mediated by increased availability of double strand breaks for integration. Additionally, kinetic analysis reveals that gene transfer (GT) events accumulate linearly as a function of time spent under antibiotic selection in the experiment, demonstrating that, contrary to what was previously thought, there is no such thing as a single GTR in somatic IGT experiments. Furthermore, IGT in tissue culture experiments appears to be the result of a "race against the clock" for integration in the nuclear genome, that starts when the organellar DNA arrives to the nucleus granting transient antibiotic resistance. GT events and escapes of kanamycin selection may be two possible outcomes from this race: those instances where the organellar DNA gets to integrate are recovered as GT events, and in those cases where timely integration fails, antibiotic resistance cannot be sustained, and end up considered as escapes. In the mutants, increased opportunities for integration in the nuclear genome change the overall ratio between IGT and escape events. The resources generated here are promising starting points for future research: (1) the mutant collection, for the further study of processes that depend on DNA repair in plants (2) the collection of GT lines obtained from these experiments, for the study of the effect of DSBR pathways over integration patterns and stability of transferred genes and (3) the developed CRISPR/Cas9 workflow for mutant generation, to make N. tabacum meet its potential as an attractive model for answering complex biological questions.
Mechanosensation is a fundamental biological process that provides the basis for sensing touch and pain as well as for hearing and proprioception. A special class of ion-channel proteins known as mechanosensitive proteins convert the mechanical stimuli into electrochemical signals to mediate this process. Mechanosensitive proteins undergo conformational changes in response to mechanical force, which eventually leads to the opening of the proteins' ion channel. Mammalian mechanosensitive proteins remained a long sought-after mystery until 2010 when a family of two proteins - Piezo1 and Piezo2 - was identifed as mechanosensors [1]. The cryo-EM structures of Piezo1 and Piezo2 protein were resolved in the last years and reveal a propeller-shaped homotrimer with 114 transmembrane helices [2, 3, 4, 5]. The protein structures are curved and have been suggested to deform the surrounding membrane into a nano-dome, which mechanically responds to membrane tension resulting from external forces [2]. In this thesis, the conformations of membrane-embedded Piezo1 and Piezo2 proteins and their tension-induced conformational changes are investigated using molecular dynamics simulations. Our coarse-grained molecular dynamics simulations show that the Piezo proteins induce curvature in the surrounding membrane and form a stable protein-membrane nano-dome in the tensionless membrane. These membrane-embedded Piezo proteins, however, adopt substantially less curved conformations in our simulations compared to the cryo-EM structures solved in detergent micelles, which agrees with recent experimental investigations of the overall Piezo nano-dome shape in membrane vesicles [6, 7, 8]. At high membrane tension, the Piezo proteins attain nearly planar conformations in our simulations. Our systematic investigation of Piezo proteins under different membrane tensions indicates a half-maximal conformational response at membrane tension values rather close to the experimentally suggested values of Piezo activation [9, 10]. In addition, our simulations indicate a widening of the Piezo1 ion channel at high membrane tension, which agrees with the channel widening observed in recent nearly flattened cryo-EM structures of Piezo1 in small membrane vesicles [11]. In contrast, the Piezo2 ion channel does not respond to membrane tension in our simulations. These different responses of the Piezo1 and Piezo2 ion channels in our simulations are in line with patch-clamp experiments, in which Piezo1, but not Piezo2, was shown to be activated by membrane tension alone [12].
The G protein-coupled estrogen receptor (GPER1) is acknowledged as an important mediator of estrogen signaling. Given the ubiquitous expression of GPER1, it is likely that the receptor plays a role in a variety of malignancies, not only in the classic hormonally regulated tissues (e.g., breast, ovary, and prostate), but also in the colon. As colorectal cancer (CRC) is the third most common cancer in both men and women worldwide and environmental factors and dietary habits are important risk factors, it is increasingly recognized that natural and synthetic hormones and their associated receptors might play a role in CRC. Through oral consumption, environmental contaminants with endocrine activity are in contact with the gastrointestinal mucosa, where they might exert their toxic effects. Although GPER1 has been shown to be engaged in physiological and pathophysiological processes, its role in CRC remains poorly understood. Thus, pro- as well as anti-tumorigenic effects are described in the literature. This thesis has uncovered novel roles of GPER1 in mediating major CRC-associated phenotypes in transformed and non-transformed colon cell lines. Exposure to the estrogens 17β-estradiol (E2), bisphenol-A (BPA) and diethylstilbestrol (DES) but also the androgen dihydrotestosterone (DHT) resulted in GPER1-dependent induction of supernumerary centrosomes, whole chromosomal instability (w-CIN) and aneuploidy. Indeed, both knockdown and inhibition of GPER1 attenuated the generation of (xeno)hormone-driven supernumerary centrosomes and karyotype instability. Mechanistically, (xeno)hormone-induced centrosome amplification was associated with transient multipolar mitosis and the generation of so called anaphase “lagging” chromosomes. The results of this thesis propose a GPER1/PKA/AKAP9-pathway in regulating centrosome numbers in colorectal cancer cells and the involvement of the centriolar protein centrin. Remarkably, exposure to (xeno)hormones resulted in atypical enlargement and unexpected phosphorylation of the centriole marker centrin in interphase. These findings provide a novel role for GPER1 in key CRC-prone lesions and shed light on underlying mechanisms that involve GPER1 function in the colon. Elucidating to what extent centrosomal proteins are involved in the GPER1-mediated aneugenic effect will be an important task for future studies. The present study was intended to lay a first foundation to understand the molecular basis and potential risk factors of CRC which might help to reduce the use of laboratory animals. Since numerous animal experiments are conducted in biomedical research, the development of alternative methods is indispensable. The Federal Institute for Risk Assessment (BfR) as the German Center for the Protection of Laboratory Animals (Bf3R) addresses this issue by uncovering underlying mechanisms leading to colorectal cancer as necessary prerequisite in order to develop alternative methods.
Plant metabolism serves as the primary mechanism for converting assimilated carbon into essential compounds crucial for plant growth and ultimately, crop yield. This renders it a focal point of research with significant implications. Despite notable strides in comprehending the genetic principles underpinning metabolism and yield, there remains a dearth of knowledge regarding the genetic factors responsible for trait variation under varying environmental conditions. Given the burgeoning global population and the advancing challenges posed by climate change, unraveling the intricacies of metabolic and yield responses to water scarcity became increasingly important in safeguarding food security.
Our research group has recently started to work on the genetic resources of legume species. To this end, the study presented here investigates the metabolic diversity across five different legume species at a tissue level, identifying species-specific biosynthesis of alkaloids as well as iso-/flavonoids with diverse functional groups, namely prenylation, phenylacylation as well as methoxylation, to create a resource for follow up studies investigation the metabolic diversity in natural diverse populations of legume species.
Following this, the second study investigates the genetic architecture of drought-induced changes in a global common bean population. Here, a plethora of quantitative trait loci (QTL) associated with various traits are identified by performing genome-wide association studies (GWAS), including for lipid signaling. On this site, overexpression of candidates highlighted the induction of several oxylipins reported to be pivotal in coping with harsh environmental conditions such as water scarcity.
Diverging from the common bean and GWAS, the following study focuses on identifying drought-related QTL in tomato using a bi-parental breeding population. This descriptive study highlights novel multi-omic QTL, including metabolism, photosynthesis as well as fruit setting, some of which are uniquely assigned under drought. Compared to conventional approaches using the bi-parental IL population, the study presented improves the resolution by assessing further backcrossed ILs, named sub-ILs.
In the final study, a photosynthetic gene, namely a PetM subunit of the cytochrome b6f complex encoding gene, involved in electron flow is characterized in an horticultural important crop. While several advances have been made in model organisms, this study highlights the transition of this fundamental knowledge to horticultural important crops, such as tomato, and investigates its function under differing light conditions. Overall, the presented thesis combines different strategies in unveiling the genetic components in multi-omic traits under drought using conventional breeding populations as well as a diverse global population. To this end, it allows a comparison of either approach and highlights their strengths and weaknesses.
Social networking sites
(2023)
Arthur Schopenhauer (1788–1860) was perhaps the last polymath among the great Germanic philosophers. Switching with ease and elegance between epistemic positions and fields as diverse as idealism and empiricism, fideism and rationalism, realism and nominalism, art and religion, jurisprudence and politics, psychology and occultism, Schopenhauer erected an imposing edifice bearing testimony to his universal learning. This study is an investigation into the very conclusion of Schopenhauer’s philosophy and endeavours to answer the following question: did Schopenhauer’s doctrine of salvation issue forth organically from his intellectual output or was it annexed to his philosophy as a result of his critical engagement with religion? The labyrinthine paths through which Schopenhauer arrives at the soteriological culmination of his philosophy are subjected to critical assessment; the picture that emerges is of a philosopher who seemed convinced that he had solved some of the most pressing cosmic riddles to have tormented mankind through the ages.
Seasonal forecasts are of great interest in many areas. Knowing the amount of precipitation for the upcoming season in regions of water scarcity would facilitate a better water management. If farmers knew the weather conditions of the upcoming summer at sowing time, they could select those cereal species that are best adapted to these conditions. This would allow farmers to improve the harvest and potentially even reduce the amount of pesticides used. However, the undoubted advantages of seasonal forecasts are often opposed by their high degree of uncertainty. The great challenge of generating seasonal forecasts with lead times of several months mainly originates from the chaotic nature of the earth system. In a chaotic system, even tiny differences in the initial conditions can lead to strong deviations in the system’s state in the long run.
In this dissertation we propose an emergent machine learning approach for seasonal forecasting, called the AnlgModel. The AnlgModel combines the analogue method with myopic feature selection and bootstrapping. To benchmark the abilities of the AnlgModel we apply it to seasonal cyclone activity forecasts in the North Atlantic and Northwest Pacific. The AnlgModel demonstrates competitive hindcast skills with two operational forecasts and even outperforms these for long lead times.
In the second chapter we comprehend the forecasting strategy of the Anlg-Model. We thereby analyse the analogue selection process for the 2017 North Atlantic and the 2018 Northwest Pacific seasonal cyclone activity. The analysis shows that those climate indices which are known to influence the seasonal cyclone activity, such as the Niño 3.4 SST, are correctly represented among the selected analogues. Furthermore the selected analogues reflect large-scale climate patterns that were identified by expert reports as being determinative for these particular seasons.
In the third chapter we analyse the features that are used by the AnlgModel for its predictions. We therefore inspect the feature relevance (FR). The FR patterns learned by the AnlgModel show a high congruence with the predictor regions used by the operational forecasts. However, the AnlgModel also discovered new features, such as the SST anomaly in the Gulf of Guinea during November. This SST pattern exhibits a remarkably high predictive potential for the upcoming Atlantic hurricane activity.
In the final chapter we investigate potential mechanisms, that link two of these regions with high feature relevance to the Atlantic hurricane activity. We mainly focus on ocean surface transport. The ocean surface flow paths are calculated using Lagrangian particle analysis. We demonstrate that the FR patterns in the region of the Canary islands do not correspond with ocean surface transport. It is instead likely that these FR patterns fingerprint a wind transport of latent heat. The second region to be studied is situated in the Gulf of Guinea. Our analysis shows that the FR patterns seen there do fingerprint ocean surface transport. However, our simulations also show that at least one other mechanism is involved in linking the Gulf of Guinea SST anomaly in November to the hurricane activity of the upcoming season.
In this work the AnlgModel does not only demonstrate its outstanding forecast skills but also shows its capabilities as research tool for detecting oceanic and atmospheric mechanisms.