Mariestad municipality participated in a global competition to become Volvo’s site for a new lithium-ion battery factory. The competition was between eleven different countries and three different locations in Sweden. Aer declaring Mariestad and the site Korstorp as winners, extensive surveys of the site were initiated during which they did a rare find of the protected species the great crested newt. However, Volvo is still planning on going through with building the factory the way they planned, which implies asphalting an area of 140-150 ha and constructing a box like factory of one or two floors. The newts are now under great threat and will have to be moved in order to make room for the factory. Building battery factories is something we strive for on a global scale, but what happens when global sustainability opposes local? In the example of the battery factory in Mariestad Agenda 2030’s sustainable development goals biodiversity and economic growth seem to be in opposition to each other and here it becomes clear that the value of humans and non-humans are different. How are we to remedy the unequal distribution of power and how can we turn the conflict zone in Korstorp into a zone of diplomacy? The convention of the rights of the child became Swedish law in 2020 and here it is relevant taking a look at article 12, which says; “All children have the right to express their opinions, adults shall listen and consider the children’s opinions”. The children are our future and they have to live with our choices. Building in a sustainable manner implies listening to who will be affected. The society’s measurement of success needs to be altered from economic growth to one which everybody has the right to be part of. To reach that place we have to practice at an early age to think and act in a democratic manner. If children feel as if they have been heard they could come to appreciate democratic processes in which they trust their ability to alter the society and feel obligated towards it. Furthermore, the unlimited imagination of children and the fact that they are not yet indoctrinated in the routines and customs of our society might bring the innovation needed to create a new kind of factory in symbiosis with the local environment.

Newts have the incredible ability to regenerate many different organs and tissues as adults, including the limbs. Limb regeneration occurs via the dedifferentiation of stump tissue and the formation of a blastema, which provides the majority of cells for the regenerate. Despite all that we have learned about dedifferentiation and blastema formation, the cellular and molecular mechanisms underlying these processes are still poorly understood. We used representational difference analysis (RDA) to identify genes involved in the early dedifferentiation process in newt forelimb regeneration. Our analysis identified approximately 410 unique genes that were differentially regulated during this process. Microarray analysis was used to determine the expression profile of these genes throughout limb and tail regeneration. We used quantitative PCR (qPCR) to validate the expression of a subset of these genes [β-catenin, wntless, dapper, thymosin-β 4 (Tβ4), and thymosin-β 10/15 (Tβ10/15)] in regenerating limb and tail tissue, as well as in differentiating newt myoblasts. We also verified the expression of these genes in the regenerating newt limb using immunohistochemistry (IHC) and in situ hybridization (ISH). Finally, we performed a functional analysis on β-catenin, wntless, dapper, and Tβ4 by overexpressing these genes in mouse myoblasts to examine their effects on differentiation and potential roles in dedifferentiation. Quantitative PCR verified the expression of β-catenin, wntless, dapper, and Tβ4 during limb regeneration and IHC/ISH localized the β-catenin and Tβ4 proteins to the blastema during regeneration. Tβ10/15 was shown by qPCR to be expressed in the tail during regeneration. Overexpression of newt β-catenin, wntless, dapper, and Tβ4 in mouse myoblasts showed that each of these genes has an inhibitory effect on the differentiation of myoblasts into myotubes and, therefore, may play a role in promoting or maintaining the dedifferentiated state. Our work has identified a large number of genes with potential roles in regulating the dedifferentiation process during newt forelimb regeneration. We have also laid a framework from which much more work can be done by drawing on the genes we have identified and the microarray data, which indicate ideal follow-up candidates. Our analysis of specific genes has also increased our understanding of the molecular events occurring during the dedifferentiation process in the regenerating newt limb.

No description available.

No description available.

In contrast to mammals, some fish and amphibians have retained the ability to regenerate complex body structures or organs, such as the limb, the tail, the eye lens or even parts of the heart. One major difference in the response to injury is the appearance of a mesenchymal growth zone or blastema in these regenerative species instead of the scarring seen in mammals. This blastema is thought to largely derive from the dedifferentiation of various functional cell types, such as skeletal muscle, skin and cartilage. In the case of multinucleated skeletal muscle fibres, cell cycle re-entry into S-phase as well as fragmentation into mononucleated progenitors is observed both in vitro and in vivo. In order to identify molecules that initiate dedifferentiation of cells at the wound site in amphibians we have established a cellular assay with a cultured newt myogenic cell line. Using this assay we have found a serum activity that stimulates cell cycle re-entry in differentiated multinucleated newt myotubes. The activity is present in serum of all mammalian species tested so far and, interestingly, thrombin proteolysis amplifies the activity from both serum and plasma. We think this serum factor provides a link between wounding and regeneration and its identification will be a key step in understanding the remarkable differences in wound healing between mammals and amphibians. In the course of this PhD thesis we have characterized the serum factor as a thermo-labile, pH- and proteinase K-sensitive, high molecular weight protein that is resistant to denaturing conditions such as SDS, urea or organic solvents. Surprisingly, under denaturing conditions the activity behaves as a low molecular weight protein that displays charge heterogeneity on isoelectric focusing. Using these characteristics of the serum factor we have performed a systematic investigation of commonly used protein chromatography modes and separation techniques to develop a successful purification procedure. After four column chromatography steps -- cation exchange, hydrophobic interaction, heparin affinity and size exclusion chromatography under denaturing conditions -- we have achieved a 2,000-fold purification starting from a commercially available Crude Bovine Thrombin preparation. This represents about 40,000-fold purification over bovine serum. Silver stained gels of the most purified fractions revealed ten major protein bands. In order to finally identify the cell cycle re-entry factor, we are currently analyzing the purification by quantitative mass spectrometry by correlating the abundance of tryptic peptides with activity in sequential fractions across a chromatography run.

This PhD study, developed at Universitat Politècnica de València (UPV), aims to cover the first phase of the benchmark released by the expert group on Uncertainty Analysis in Modeling (UAM-LWR). The main contribution to the benchmark, made by the thesis' author, is the development of a MATLAB program requested by the benchmark organizers. This is used to generate neutronic libraries to distribute among the benchmark participants. The UAM benchmark pretends to determine the uncertainty introduced by coupled multi-physics and multi-scale LWR analysis codes. The benchmark is subdivided into three phases: 1. Neutronic phase: obtain collapsed and homogenized problem-dependent cross sections and criticality analyses. 2. Core phase: standalone thermohydraulic and neutronic codes. 3. System phase: coupled thermohydraulic and neutronic code. In this thesis the objectives of the first phase are covered. Specifically, a methodology is developed to propagate the uncertainty of cross sections and other neutronic parameters through a lattice physics code and core simulator. An Uncertainty and Sensitivity (U&S) analysis is performed over the cross sections contained in the ENDF/B-VII nuclear library. Their uncertainty is propagated through the lattice physics code SCALE6.2.1, including the collapse and homogenization phase, up to the generation of problem-dependent neutronic libraries. Afterward, the uncertainty contained in these libraries can be further propagated through a core simulator, in this study PARCSv3.2. The module SAMPLER -available in the latest release of SCALE- and DAKOTA 6.3 statistical tool are used for the U&S analysis. As a part of this process, a methodology to obtain neutronic libraries in NEMTAB format -to be used in a core simulator- is also developed. A code-to-code comparison with CASMO-4 is used as a verification. The whole methodology is tested using a Boiling Water Reactor (BWR) reactor type. Nevertheless, there is not any concern or limitation regarding its use in any other type of nuclear reactor. The Gesellschaft für Anlagen und Reaktorsicherheit (GRS) stochastic methodology for uncertainty quantification is used. This methodology makes use of the high-fidelity model and nonparametric sampling to propagate the uncertainty. As a result, the number of samples (determined using the revised Wilks' formula) does not depend on the number of input parameters but only on the desired confidence and uncertainty of output parameters. Moreover, the output Probability Distribution Functions (PDFs) are not subject to normality. The main disadvantage is that each input parameter must have a pre-defined PDF. If possible, input PDFs are defined using information found in the related literature. Otherwise, the uncertainty definition is based on expert judgment. A second scenario is used to propagate the uncertainty of different thermohydraulic parameters through the coupled code TRACE5.0p3/PARCSv3.0. In this case, a PWR reactor type is used and a transient control rod drop occurrence is simulated. As a new feature, the core is modeled chan-by-chan following a fully 3D discretization. No other study is found using a detailed 3D core. This U&S analysis also makes use of the GRS methodology and DAKOTA 6.3. / Este trabajo de doctorado, desarrollado en la Universitat Politècnica de València (UPV), tiene como objetivo cubrir la primera fase del benchmark presentado por el grupo de expertos Uncertainty Analysis in Modeling (UAM-LWR). La principal contribución al benchmark, por parte del autor de esta tesis, es el desarrollo de un programa de MATLAB solicitado por los organizadores del benchmark, el cual se usa para generar librerías neutrónicas a distribuir entre los participantes del benchmark. El benchmark del UAM pretende determinar la incertidumbre introducida por los códigos multifísicos y multiescala acoplados de análisis de reactores de agua ligera. El citado benchmark se divide en tres fases: 1. Fase neutrónica: obtener los parámetros neutrónicos y secciones eficaces del problema específico colapsados y homogenizados, además del análisis de criticidad. 2. Fase de núcleo: análisis termo-hidráulico y neutrónico por separado. 3. Fase de sistema: análisis termo-hidráulico y neutrónico acoplados. En esta tesis se completan los principales objetivos de la primera fase. Concretamente, se desarrolla una metodología para propagar la incertidumbre de secciones eficaces y otros parámetros neutrónicos a través de un código lattice y un simulador de núcleo. Se lleva a cabo un análisis de incertidumbre y sensibilidad para las secciones eficaces contenidas en la librería neutrónica ENDF/B-VII. Su incertidumbre se propaga a través del código lattice SCALE6.2.1, incluyendo las fases de colapsación y homogenización, hasta llegar a la generación de una librería neutrónica específica del problema. Luego, la incertidumbre contenida en dicha librería puede continuar propagándose a través de un simulador de núcleo, para este estudio PARCSv3.2. Para el análisis de incertidumbre y sensibilidad se ha usado el módulo SAMPLER -disponible en la última versión de SCALE- y la herramienta estadística DAKOTA 6.3. Como parte de este proceso, también se ha desarrollado una metodología para obtener librerías neutrónicas en formato NEMTAB para ser usadas en simuladores de núcleo. Se ha realizado una comparación con el código CASMO-4 para obtener una verificación de la metodología completa. Esta se ha probado usando un reactor de agua en ebullición del tipo BWR. Sin embargo, no hay ninguna preocupación o limitación respecto a su uso con otro tipo de reactor nuclear. Para la cuantificación de la incertidumbre se usa la metodología estocástica Gesellschaft für Anlagen und Reaktorsicherheit (GRS). Esta metodología hace uso del modelo de alta fidelidad y un muestreo no paramétrico para propagar la incertidumbre. Como resultado, el número de muestras (determinado con la fórmula revisada de Wilks) no depende del número de parámetros de entrada, sólo depende del nivel de confianza e incertidumbre deseados de los parámetros de salida. Además, las funciones de distribución de probabilidad no están limitadas a normalidad. El principal inconveniente es que se ha de disponer de las distribuciones de probabilidad de cada parámetro de entrada. Si es posible, las distribuciones de probabilidad de entrada se definen usando información encontrada en la literatura relacionada. En caso contrario, la incertidumbre se define en base a la opinión de un experto. Se usa un segundo escenario para propagar la incertidumbre de diferentes parámetros termo-hidráulicos a través del código acoplado TRACE5.0p3/PARCSv3.0. En este caso, se utiliza un reactor tipo PWR para simular un transitorio de una caída de barra. Como nueva característica, el núcleo se modela elemento a elemento siguiendo una discretización totalmente en 3D. No se ha encontrado ningún otro estudio que use un núcleo tan detallado en 3D. También se usa la metodología GRS y el DAKOTA 6.3 para este análisis de incertidumbre y sensibilidad. / Aquest treball de doctorat, desenvolupat a la Universitat Politècnica de València (UPV), té com a objectiu cobrir la primera fase del benchmark presentat pel grup d'experts Uncertainty Analysis in Modeling (UAM-LWR). La principal contribució al benchmark, per part de l'autor d'aquesta tesi, es el desenvolupament d'un programa de MATLAB sol¿licitat pels organitzadors del benchmark, el qual s'utilitza per a generar llibreries neutròniques a distribuir entre els participants del benchmark. El benchmark del UAM pretén determinar la incertesa introduïda pels codis multifísics i multiescala acoblats d'anàlisi de reactors d'aigua lleugera. El citat benchmark es divideix en tres fases: 1. Fase neutrònica: obtenir els paràmetres neutrònics i seccions eficaces del problema específic, col¿lapsats i homogeneïtzats, a més de la anàlisi de criticitat. 2. Fase de nucli: anàlisi termo-hidràulica i neutrònica per separat. 3. Fase de sistema: anàlisi termo-hidràulica i neutrònica acoblats. En aquesta tesi es completen els principals objectius de la primera fase. Concretament, es desenvolupa una metodologia per propagar la incertesa de les seccions eficaces i altres paràmetres neutrònics a través d'un codi lattice i un simulador de nucli. Es porta a terme una anàlisi d'incertesa i sensibilitat per a les seccions eficaces contingudes en la llibreria neutrònica ENDF/B-VII. La seua incertesa es propaga a través del codi lattice SCALE6.2.1, incloent les fases per col¿lapsar i homogeneïtzar, fins aplegar a la generació d'una llibreria neutrònica específica del problema. Després, la incertesa continguda en la esmentada llibreria pot continuar propagant-se a través d'un simulador de nucli, per a aquest estudi PARCSv3.2. Per a l'anàlisi d'incertesa i sensibilitat s'ha utilitzat el mòdul SAMPLER -disponible a l'última versió de SCALE- i la ferramenta estadística DAKOTA 6.3. Com a part d'aquest procés, també es desenvolupa una metodologia per a obtenir llibreries neutròniques en format NEMTAB per ser utilitzades en simuladors de nucli. S'ha realitzat una comparació amb el codi CASMO-4 per obtenir una verificació de la metodologia completa. Aquesta s'ha provat utilitzant un reactor d'aigua en ebullició del tipus BWR. Tanmateix, no hi ha cap preocupació o limitació respecte del seu ús amb un altre tipus de reactor nuclear. Per a la quantificació de la incertesa s'utilitza la metodologia estocàstica Gesellschaft für Anlagen und Reaktorsicherheit (GRS). Aquesta metodologia fa ús del model d'alta fidelitat i un mostreig no paramètric per propagar la incertesa. Com a resultat, el nombre de mostres (determinat amb la fórmula revisada de Wilks) no depèn del nombre de paràmetres d'entrada, sols depèn del nivell de confiança i incertesa desitjats dels paràmetres d'eixida. A més, las funcions de distribució de probabilitat no estan limitades a la normalitat. El principal inconvenient és que s'ha de disposar de les distribucions de probabilitat de cada paràmetre d'entrada. Si és possible, les distribucions de probabilitat d'entrada es defineixen utilitzant informació trobada a la literatura relacionada. En cas contrari, la incertesa es defineix en base a l'opinió d'un expert. S'utilitza un segon escenari per propagar la incertesa de diferents paràmetres termo-hidràulics a través del codi acoblat TRACE5.0p3/PARCSv3.0. En aquest cas, s'utilitza un reactor tipus PWR per simular un transitori d'una caiguda de barra. Com a nova característica, cal assenyalar que el nucli es modela element a element seguint una discretizació totalment 3D. No s'ha trobat cap altre estudi que utilitze un nucli tan detallat en 3D. També s'utilitza la metodologia GRS i el DAKOTA 6.3 per a aquesta anàlisi d'incertesa i sensibilitat.¿ / Mesado Melia, C. (2017). Uncertainty Quantification and Sensitivity Analysis for Cross Sections and Thermohydraulic Parameters in Lattice and Core Physics Codes. Methodology for Cross Section Library Generation and Application to PWR and BWR [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/86167

In contrast to mammals, some fish and amphibians have retained the ability to regenerate complex body structures or organs, such as the limb, the tail, the eye lens or even parts of the heart. One major difference in the response to injury is the appearance of a mesenchymal growth zone or blastema in these regenerative species instead of the scarring seen in mammals. This blastema is thought to largely derive from the dedifferentiation of various functional cell types, such as skeletal muscle, skin and cartilage. In the case of multinucleated skeletal muscle fibres, cell cycle re-entry into S-phase as well as fragmentation into mononucleated progenitors is observed both in vitro and in vivo. In order to identify molecules that initiate dedifferentiation of cells at the wound site in amphibians we have established a cellular assay with a cultured newt myogenic cell line. Using this assay we have found a serum activity that stimulates cell cycle re-entry in differentiated multinucleated newt myotubes. The activity is present in serum of all mammalian species tested so far and, interestingly, thrombin proteolysis amplifies the activity from both serum and plasma. We think this serum factor provides a link between wounding and regeneration and its identification will be a key step in understanding the remarkable differences in wound healing between mammals and amphibians. In the course of this PhD thesis we have characterized the serum factor as a thermo-labile, pH- and proteinase K-sensitive, high molecular weight protein that is resistant to denaturing conditions such as SDS, urea or organic solvents. Surprisingly, under denaturing conditions the activity behaves as a low molecular weight protein that displays charge heterogeneity on isoelectric focusing. Using these characteristics of the serum factor we have performed a systematic investigation of commonly used protein chromatography modes and separation techniques to develop a successful purification procedure. After four column chromatography steps -- cation exchange, hydrophobic interaction, heparin affinity and size exclusion chromatography under denaturing conditions -- we have achieved a 2,000-fold purification starting from a commercially available Crude Bovine Thrombin preparation. This represents about 40,000-fold purification over bovine serum. Silver stained gels of the most purified fractions revealed ten major protein bands. In order to finally identify the cell cycle re-entry factor, we are currently analyzing the purification by quantitative mass spectrometry by correlating the abundance of tryptic peptides with activity in sequential fractions across a chromatography run.

No description available.