Der Einfluss assistierter Reproduktionstechniken auf die epigenetische Reprogrammierung in der fruehen Embryogenese und Gametogenese im Mausmodell

Seit der Geburt von Louise J. Brown (1978) als erstem künstlich erzeugtem Kind hat sich die Nachfrage nach assistierten Reproduktionstechniken (ART) stark erhöht. Der Anteil der nach In-vitro-Fertilisation (IVF) oder Intrazytoplasmatischer Spermieninjektion (ICSI) geborenen Kinder macht mittlerweile abhängig vom betrachteten Industrieland zwischen 1-4% an der Gesamtgeburtenzahl aus. In zahlreichen Studien korreliert eine erhöhte Prävalenz für seltene Imprinting-Erkrankungen, wie z.B. Beckwith-Wiedemann oder Angelman-Syndrom, mit der Geburt nach assistierten Reproduktionstechniken. Es ist bekannt, dass die medizinischen Interventionen zur Behandlung von Sub- und Infertilität in sehr sensitive Phasen der epigenetischen Reprogrammierung des Embryos und der Keimzellen eingreifen. In der vorliegenden Arbeit wurde untersucht, ob die ovarielle Stimulation einen Einfluss auf die epigenetische Integrität von geprägten Genen in murinen Präimplantationsembryonen hat. Die in diesem Zusammenhang entwickelte digitale Bisulfitpyrosequenzierung gewährleistet die Analyse der DNA-Methylierung auf Einzelallelebene durch eine adäquate Verdünnung der Probe im Vorfeld der PCR. Die ovarielle Induktion führte zu einem erhöhten Rate an Epimutationen des paternalen H19-Allels, sowie des maternalen Snrpn-Allels. Zudem konnte festgestellt werden, dass die Expression von drei potentiellen Reprogrammierungsgenen (Apex1, Polb, Mbd3) in Embryonen aus hormonell stimulierten Muttertieren dereguliert ist. Whole-Mount Immunfluoreszenzfärbungen für APEX1 korrelierten dessen differentielle Genexpression mit dem Proteinlevel. Anzeichen früher apoptotischer Vorgänge äußerten sich in Embryonen aus hormonell induzierten Muttertieren in der hohen Rate an Embryonen, die keines der drei Transkripte exprimierten oder weniger APEX1-positive Blastomeren aufwiesen.In einer weiteren Fragestellung wurde untersucht, ob die Kryokonservierung muriner Spermatozoen den epigenetischen Status geprägter Gene in den Keimzellen beeinflusst. Die Analyse von F1-Zweizellembryonen, die durch IVF mit den jeweiligen Spermatozoen eines Männchens generiert wurden, diente der Aufklärung möglicher paternaler Transmissionen. Insgesamt konnten keine signifikanten Auswirkungen der Kryokonservierung auf den epigenetischen Status in Spermatozoen und F1-Embryonen ermittelt werden.

Direct and inverse transient eddy current problems

Die vorliegende Arbeit behandelt Vorwärts- sowie Rückwärtstheorie transienter Wirbelstromprobleme. Transiente Anregungsströme induzieren elektromagnetische Felder, welche sogenannte Wirbelströme in leitfähigen Objekten erzeugen. Im Falle von sich langsam ändernden Feldern kann diese Wechselwirkung durch die Wirbelstromgleichung, einer Approximation an die Maxwell-Gleichungen, beschrieben werden. Diese ist eine lineare partielle Differentialgleichung mit nicht-glatten Koeffizientenfunktionen von gemischt parabolisch-elliptischem Typ. Das Vorwärtsproblem besteht darin, zu gegebener Anregung sowie den umgebungsbeschreibenden Koeffizientenfunktionen das elektrische Feld als distributionelle Lösung der Gleichung zu bestimmen. Umgekehrt können die Felder mit Messspulen gemessen werden. Das Ziel des Rückwärtsproblems ist es, aus diesen Messungen Informationen über leitfähige Objekte, also über die Koeffizientenfunktion, die diese beschreibt, zu gewinnen. In dieser Arbeit wird eine variationelle Lösungstheorie vorgestellt und die Wohlgestelltheit der Gleichung diskutiert. Darauf aufbauend wird das Verhalten der Lösung für verschwindende Leitfähigkeit studiert und die Linearisierbarkeit der Gleichung ohne leitfähiges Objekt in Richtung des Auftauchens eines leitfähigen Objektes gezeigt. Zur Regularisierung der Gleichung werden Modifikationen vorgeschlagen, welche ein voll parabolisches bzw. elliptisches Problem liefern. Diese werden verifiziert, indem die Konvergenz der Lösungen gezeigt wird. Zuletzt wird gezeigt, dass unter der Annahme von sonst homogenen Umgebungsparametern leitfähige Objekte eindeutig durch die Messungen lokalisiert werden können. Hierzu werden die Linear Sampling Methode sowie die Faktorisierungsmethode angewendet.

The Feynman-Kac formula and applications to PDEs

The thesis presents a probabilistic approach to the theory of semigroups of operators, with particular attention to the Markov and Feller semigroups. The first goal of this work is the proof of the fundamental Feynman-Kac formula, which gives the solution of certain parabolic Cauchy problems, in terms of the expected value of the initial condition computed at the associated stochastic diffusion processes. The second target is the characterization of the principal eigenvalue of the generator of a semigroup with Markov transition probability function and of second order elliptic operators with real coefficients not necessarily self-adjoint. The thesis is divided into three chapters. In the first chapter we study the Brownian motion and some of its main properties, the stochastic processes, the stochastic integral and the Itô formula in order to finally arrive, in the last section, at the proof of the Feynman-Kac formula. The second chapter is devoted to the probabilistic approach to the semigroups theory and it is here that we introduce Markov and Feller semigroups. Special emphasis is given to the Feller semigroup associated with the Brownian motion. The third and last chapter is divided into two sections. In the first one we present the abstract characterization of the principal eigenvalue of the infinitesimal generator of a semigroup of operators acting on continuous functions over a compact metric space. In the second section this approach is used to study the principal eigenvalue of elliptic partial differential operators with real coefficients. At the end, in the appendix, we gather some of the technical results used in the thesis in more details. Appendix A is devoted to the Sion minimax theorem, while in appendix B we prove the Chernoff product formula for not necessarily self-adjoint operators.

Distinct DNA methylation patterns in cirrhotic liver and hepatocellular carcinoma

Abberrant DNA methylation is one of the hallmarks of cancerogenesis. Our study aims to delineate differential DNA methylation in cirrhosis and hepatic cancerogenesis. Patterns of methylation of 27,578 individual CpG loci in 12 hepatocellular carcinomas (HCCs), 15 cirrhotic controls and 12 normal liver samples were investigated using an array-based technology. A supervised principal component analysis (PCA) revealed 167 hypomethylated loci and 100 hypermethylated loci in cirrhosis and HCC as compared to normal controls. Thus, these loci show a "cirrhotic" methylation pattern that is maintained in HCC. In pairwise supervised PCAs between normal liver, cirrhosis and HCC, eight loci were significantly changed in all analyses differentiating the three groups (p < 0.0001). Of these, five loci showed highest methylation levels in HCC and lowest in control tissue (LOC55908, CELSR1, CRMP1, GNRH2, ALOX12 and ANGPTL7), whereas two loci showed the opposite direction of change (SPRR3 and TNFSF15). Genes hypermethylated between normal liver to cirrhosis, which maintain this methylation pattern during the development of HCC, are depleted for CpG islands, high CpG content promoters and polycomb repressive complex 2 (PRC2) targets in embryonic stem cells. In contrast, genes selectively hypermethylated in HCC as compared to nonmalignant samples showed an enrichment of CpG islands, high CpG content promoters and PRC2 target genes (p < 0.0001). Cirrhosis and HCC show distinct patterns of differential methylation with regards to promoter structure, PRC2 targets and CpG islands.

Secretin receptors in the human liver: expression in biliary tract and cholangiocarcinoma, but not in hepatocytes or hepatocellular carcinoma

BACKGROUND/AIMS: Gut hormone receptors are over-expressed in human cancer and allow receptor-targeted tumor imaging and therapy. A novel promising receptor for these purposes is the secretin receptor. The secretin receptor expression was investigated in the human liver because the liver is a physiological secretin target and because novel diagnostic and treatment modalities are needed for liver cancer. METHODS: Nineteen normal livers, 10 cirrhotic livers, 35 cholangiocarcinomas, and 45 hepatocellular carcinomas were investigated for secretin receptor expression by in vitro receptor autoradiography using (125)I-[Tyr(10)] rat secretin and, in selected cases, for secretin receptor mRNA by RT-PCR. RESULTS: Secretin receptors were present in normal bile ducts and ductules, but not in hepatocytes. A significant receptor up-regulation was observed in ductular reaction in liver cirrhosis. Twenty-two (63%) cholangiocarcinomas were positive for secretin receptors, while hepatocellular carcinomas were negative. RT-PCR revealed wild-type receptor mRNA in the non-neoplastic liver, wild-type and spliced variant receptor mRNAs in cholangiocarcinomas found receptor positive in autoradiography experiments, and no receptor transcripts in autoradiographically negative cholangiocarcinomas. CONCLUSIONS: The expression of secretin receptors in the biliary tract is the molecular basis of the secretin-induced bicarbonate-rich choleresis in man. The high receptor expression in cholangiocarcinomas may be used for in vivo secretin receptor-targeting of these tumors and for the differential diagnosis with hepatocellular carcinoma.

The Structure and Correlates of Self-Reported DSM-5 Maladaptive Personality Traits: Findings From Two German-Speaking Samples

The authors investigated the structure and correlates of DSM-5 maladaptive personality traits in two samples of 577 students and 212 inpatients using the German self-report form of the Personality Inventory for DSM-5. They found that (a) the factor structure of DSM-5 trait facets is largely in line with the proposed trait domains of Negative Affectivity, Detachment, Antagonism, Disinhibition, and Psychoticism; (b) all DSM-5 trait domains except Psychoticism are highly related to the respective domains of the Five-Factor Model of personality; (c) the trait facets are positively associated with a self-report measure of general personality dysfunction; and (d) the DSM-5 trait facets show differential associations with a range of self-reported DSM-IV Axis I disorders. These findings give further support to the new DSM-5 trait model and suggest that it may generalize to other languages and cul

Cultura, identidad y territorio : Reflexiones para el desarrollo de una estrategia de diferenciación del servicio agroturístico en emprendimientos de un Grupo de Cambio Rural (INTA) del partido de Coronel Suárez (Buenos Aires)

A partir de los conceptos convergentes identidad, cultura, patrimonio y territorio, se plantea analizar la posibilidad de formular e implementar una estrategia de diferenciación para la actividad de agroturismo desarrollada por los establecimientos del grupo Cortaderas II (Cambio Rural, INTA), en el partido de Coronel Suárez (BA), provincia de Buenos Aires. Con este fin, se ha efectuado un diagnóstico mediante entrevistas directas y observaciones in situ de los emprendimientos participantes. Destacando aspectos propios de su ambiente rural, se procuró identificar valores culturales y productos con anclaje territorial que impongan un sello distintivo al servicio turístico que ofrecen. Complementariamente, se indagó en qué medida los integrantes están preparados para gestionar el negocio, reconociendo en ellos competencias gerenciales y características emprendedoras. Por último, se reflexiona acerca del grado de madurez alcanzado, tipificando la cultura empresarial individual y según el trabajo grupal, de modo de evaluar si la puesta en valor de sus recursos evoluciona desde un "carácter doméstico" hasta llegar a un "carácter comercial formal". Se observa la construcción de una identidad de índole socio-cultural-económica que hace viable una estrategia competitiva diferencial con anclaje en el territorio, así como condiciones para el desarrollo de una actividad turística más formalizada basada en la gestión de calidad

Cultura, identidad y territorio : Reflexiones para el desarrollo de una estrategia de diferenciación del servicio agroturístico en emprendimientos de un Grupo de Cambio Rural (INTA) del partido de Coronel Suárez (Buenos Aires)

A partir de los conceptos convergentes identidad, cultura, patrimonio y territorio, se plantea analizar la posibilidad de formular e implementar una estrategia de diferenciación para la actividad de agroturismo desarrollada por los establecimientos del grupo Cortaderas II (Cambio Rural, INTA), en el partido de Coronel Suárez (BA), provincia de Buenos Aires. Con este fin, se ha efectuado un diagnóstico mediante entrevistas directas y observaciones in situ de los emprendimientos participantes. Destacando aspectos propios de su ambiente rural, se procuró identificar valores culturales y productos con anclaje territorial que impongan un sello distintivo al servicio turístico que ofrecen. Complementariamente, se indagó en qué medida los integrantes están preparados para gestionar el negocio, reconociendo en ellos competencias gerenciales y características emprendedoras. Por último, se reflexiona acerca del grado de madurez alcanzado, tipificando la cultura empresarial individual y según el trabajo grupal, de modo de evaluar si la puesta en valor de sus recursos evoluciona desde un "carácter doméstico" hasta llegar a un "carácter comercial formal". Se observa la construcción de una identidad de índole socio-cultural-económica que hace viable una estrategia competitiva diferencial con anclaje en el territorio, así como condiciones para el desarrollo de una actividad turística más formalizada basada en la gestión de calidad

Cultura, identidad y territorio : Reflexiones para el desarrollo de una estrategia de diferenciación del servicio agroturístico en emprendimientos de un Grupo de Cambio Rural (INTA) del partido de Coronel Suárez (Buenos Aires)

A partir de los conceptos convergentes identidad, cultura, patrimonio y territorio, se plantea analizar la posibilidad de formular e implementar una estrategia de diferenciación para la actividad de agroturismo desarrollada por los establecimientos del grupo Cortaderas II (Cambio Rural, INTA), en el partido de Coronel Suárez (BA), provincia de Buenos Aires. Con este fin, se ha efectuado un diagnóstico mediante entrevistas directas y observaciones in situ de los emprendimientos participantes. Destacando aspectos propios de su ambiente rural, se procuró identificar valores culturales y productos con anclaje territorial que impongan un sello distintivo al servicio turístico que ofrecen. Complementariamente, se indagó en qué medida los integrantes están preparados para gestionar el negocio, reconociendo en ellos competencias gerenciales y características emprendedoras. Por último, se reflexiona acerca del grado de madurez alcanzado, tipificando la cultura empresarial individual y según el trabajo grupal, de modo de evaluar si la puesta en valor de sus recursos evoluciona desde un "carácter doméstico" hasta llegar a un "carácter comercial formal". Se observa la construcción de una identidad de índole socio-cultural-económica que hace viable una estrategia competitiva diferencial con anclaje en el territorio, así como condiciones para el desarrollo de una actividad turística más formalizada basada en la gestión de calidad

Estimation of cooling of lavas from Serocki volcano

A dynamic crystallization study was undertaken to provide a framework for linking the textural variations observed in the Hole 648B lavas with the size and morphology of cooling units inferred from drilling and submersible observation. The textures produced in cooling rate experiments carried out using a Serocki lava (ALV-1690-20) are comparable to the groundmass textural characteristics of lavas from Serocki volcano. The results of the dynamic crystallization study provide a quantitative link between texture, cooling rate, and eruption temperature. The maximum half-width of cooling units estimated from textural characteristics is on the order of 3 m, a value consistent with constraints from drilling and submersible observation. Textural characteristics indicate that the temperature from which cooling began was slightly above the liquidus. The relation between cooling rate and texture are also tested on a drill core sample of basalt of similar composition from a 9-m-thick flow in DSDP Hole 396B.

On the non-slip boundary condition enforcement in SPH methods.

The implementation of boundary conditions is one of the points where the SPH methodology still has some work to do. The aim of the present work is to provide an in-depth analysis of the most representative mirroring techniques used in SPH to enforce boundary conditions (BC) along solid profiles. We specifically refer to dummy particles, ghost particles, and Takeda et al. [1] boundary integrals. A Pouseuille flow has been used as a example to gradually evaluate the accuracy of the different implementations. Our goal is to test the behavior of the second-order differential operator with the proposed boundary extensions when the smoothing length h and other dicretization parameters as dx/h tend simultaneously to zero. First, using a smoothed continuous approximation of the unidirectional Pouseuille problem, the evolution of the velocity profile has been studied focusing on the values of the velocity and the viscous shear at the boundaries, where the exact solution should be approximated as h decreases. Second, to evaluate the impact of the discretization of the problem, an Eulerian SPH discrete version of the former problem has been implemented and similar results have been monitored. Finally, for the sake of completeness, a 2D Lagrangian SPH implementation of the problem has been also studied to compare the consequences of the particle movement

On the flexural vibration of cylinders under axial loads:Numerical and experimental study

The flexural vibration of a homogeneous isotropic linearly elastic cylinder of any aspect ratio is analysed in this paper. Natural frequencies of a cylinder under uniformly distributed axial loads acting on its bases are calculated numerically by the Ritz method with terms of power series in the coordinate directions as approximating functions. The effect of axial loads on the flexural vibration cannot be described by applying infinitesimal strain theory, therefore, geometrically nonlinear strain–displacement relations with second-order terms are considered here. The natural frequencies of free–free, clamped–clamped, and sliding–sliding cylinders subjected to axial loads are calculated using the proposed three-dimensional Ritz approach and are compared with those obtained with the finite element method and the Bernoulli–Euler theory. Different experiments with cylinders axially compressed by a hydraulic press are carried out and the experimental results for the lowest flexural frequency are compared with the numerical results. An approach based on the Ritz formulation is proposed for the flexural vibration of a cylinder between the platens of the press with constraints varying with the intensity of the compression. The results show that for low compressions the cylinder behaves similarly to a sliding–sliding cylinder, whereas for high compressions the cylinder vibrates as a clamped–clamped one.

Simulación numérica computacional de fenómenos dinámicos

En este proyecto se trata la simulación numérica de un fenómeno dinámico, basado en el comportamiento de una onda transmitida a lo largo de una cuerda elástica de un instrumento musical, cuyos extremos se encuentran anclados. El fenómeno físico, se desarrolla utilizando una ecuación en derivadas parciales hiperbólicas con variables espacial y temporal, acompañada por unas condiciones de contorno tipo Dirichlet en los extremos y por más condiciones iniciales que dan comienzo al proceso. Posteriormente se han generado algoritmos para el método numérico empleado (Diferencias finitas centrales y progresivas) y la programación del problema aproximado con su consistencia, estabilidad y convergencia, obteniéndose unos resultados acordes con la solución analítica del problema matemático. La programación y salida de resultados se ha realizado con Visual Studio 8.0. y la programación de objetos con Visual Basic .Net In this project the topic is the numerical simulation of a dynamic phenomenon, based on the behavior of a transmitted wave along an elastic string of a musical instrument, whose ends are anchored. The physical phenomenon is developed using a hyperbolic partial differential equation with spatial and temporal variables, accompanied by a Dirichlet boundary conditions at the ends and more initial conditions that start the process. Subsequently generated algorithms for the numerical method used (central and forward finite differences) and the programming of the approximate problem with consistency, stability and convergence, yielding results in line with the analytical solution of the mathematical problem. Programming and output results has been made with Visual Studio 8.0. and object programming with Visual Basic. Net

Design of a Semi-analytical Method to Describe Plasma-Wall Interactions

Electric probes are objects immersed in the plasma with sharp boundaries which collect of emit charged particles. Consequently, the nearby plasma evolves under abrupt imposed and/or naturally emerging conditions. There could be localized currents, different time scales for plasma species evolution, charge separation and absorbing-emitting walls. The traditional numerical schemes based on differences often transform these disparate boundary conditions into computational singularities. This is the case of models using advection-diffusion differential equations with source-sink terms (also called Fokker-Planck equations). These equations are used in both, fluid and kinetic descriptions, to obtain the distribution functions or the density for each plasma species close to the boundaries. We present a resolution method grounded on an integral advancing scheme by using approximate Green's functions, also called short-time propagators. All the integrals, as a path integration process, are numerically calculated, what states a robust grid-free computational integral method, which is unconditionally stable for any time step. Hence, the sharp boundary conditions, as the current emission from a wall, can be treated during the short-time regime providing solutions that works as if they were known for each time step analytically. The form of the propagator (typically a multivariate Gaussian) is not unique and it can be adjusted during the advancing scheme to preserve the conserved quantities of the problem. The effects of the electric or magnetic fields can be incorporated into the iterative algorithm. The method allows smooth transitions of the evolving solutions even when abrupt discontinuities are present. In this work it is proposed a procedure to incorporate, for the very first time, the boundary conditions in the numerical integral scheme. This numerical scheme is applied to model the plasma bulk interaction with a charge-emitting electrode, dealing with fluid diffusion equations combined with Poisson equation self-consistently. It has been checked the stability of this computational method under any number of iterations, even for advancing in time electrons and ions having different time scales. This work establishes the basis to deal in future work with problems related to plasma thrusters or emissive probes in electromagnetic fields.

Molecular organization of the postsynaptic specialization

A specific set of molecules including glutamate receptors is targeted to the postsynaptic specialization of excitatory synapses in the brain, gathering in a structure known as the postsynaptic density (PSD). Synaptic targeting of glutamate receptors depends on interactions between the C-terminal tails of receptor subunits and specific PDZ domain-containing scaffold proteins in the PSD. These scaffold proteins assemble a specialized protein complex around each class of glutamate receptor that functions in signal transduction, cytoskeletal anchoring, and trafficking of the receptors. Among the glutamate receptor subtypes, the N-methyl-d-aspartate receptor is relatively stably integrated in the PSD, whereas the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor moves in and out of the postsynaptic membrane in highly dynamic fashion. The distinctive cell biological behaviors of N-methyl-d-aspartate and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors can be explained by their differential interactions with cytoplasmic proteins.