No description available.

No description available.

<p>Germ cells represent the immortal line: they are guardians of a totipotent genome and are essential for the genetic survival of an individual organism and ultimately a species. An error at any stage in development (specification, migration, colonization, differentiation, adult maintenance) can lead to one of two disastrous outcomes: (1) germ cell death or (2) unchecked growth and proliferation leading to tumorigenesis. The work in this dissertation utilizes a classic mouse model (<italic>Ter</italic>) resulting in both of these phenotypes to further explore the molecular mechanisms important for development of germ cells. </p> <p>A homozygous nonsense mutation (<italic>Ter</italic>) in murine <italic>Dnd1</italic> (<italic>Dnd1<super>Ter/Ter</super></italic>) results in a significant (but not complete) early loss of primordial germ cells (PGCs) prior to colonization of the gonad in both sexes and all genetic backgrounds tested. The same mutation also leads to testicular teratomas only on the 129/SvJ background. Male mutants on other genetic backgrounds ultimately lose all PGCs with no incidence of teratoma formation. It is not clear how these PGCs are lost, develop into teratomas, or what factors directly control the strain-specific phenotype variation. </p> <p>Work here demonstrates that <italic>Dnd1</italic> expression is restricted to germ cells and that the <italic>Ter</italic> mutant defect is cell autonomous. The early loss of germ cells is due in part to BAX&ndash;mediated apoptosis which also affects the incidence of tumorigenesis on a mixed genetic background. Moreover, tumor formation is-specific to the male developmental pathway and not dependent on sex chromosome composition of the germ cell (XX vs. XY). Despite normal initiation of the male somatic pathway, mutant germ cells fail to differentiate as pro&ndash;spermatogonia and instead prematurely enter meiosis.</p> <p>Results here also reveal that, on a 129/SvJ background, many mutant germ cells fail to commit to the male differentiation pathway, instead maintain expression of the pluripotency markers, NANOG, SOX2, and OCT4, and initiate teratoma formation at the stage when male germ cells normally enter mitotic arrest. RNA immunoprecipitation experiments reveal that mouse DND1 directly binds a group of transcripts that encode negative regulators of the cell cycle, including <italic>p27Kip1</italic>, which is not translated in <italic>Dnd1<super>Ter/Ter</super></italic> germ cells. Additionally, overexpression of DND1 in a teratocarcinoma cell line leads to significant alteration of pathways controlling the G1/S checkpoint and the RB tumor suppressor protein. This strongly suggests that DND1 regulates mitotic arrest in male germ cells through regulation of cell cycle genes, serving as a gatekeeper to prevent the activation of a pluripotent program leading to teratoma formation. Furthermore, strain&ndash;specific morphological and expression level differences possibly account for sensitivity to tumor development.</p> / Dissertation

The annular-return flow and the utility of small-scale, combustion-based actuators for its control are investigated experimentally. The annular return flow is generated by an axial primary round jet, which impinges normally on a bounded end wall of a concentric tube, subsequently reverses direction, and exits the tube in a countercurrent flow to the primary jet. The combustion-based actuator generates a momentary (pulsed) jet that is produced by the ignition of a mixture of gaseous fuel and oxidizer in a small (cubic centimeter scale) combustion chamber. The operating frequency and the phase can be continuously varied by independently controlling the flow rate of the fuel/oxidizer and the ignition frequency. Two radially-opposing actuators are mounted on the wall of the annular return tube and are used to trigger flow transients that alter the global flow through strong feedback. The characteristics of the baseline flow and the effects of actuation are investigated using particle image velocimetry (PIV) as well as static and unsteady pressure measurements. The baseline flow is highly unstable, exhibiting very high rates of flow recirculation. The actuator jet acts as an azimuthal obstruction deflecting the primary jet and causing it to flow around the actuator jet. Furthermore, the interaction of the primary jet with the actuator jets generates large-scale circulation domains.

The induction of germ-line chimerism is an expanding focus of fisheries research. This technique is having a potential to enhance the production of gametes of species that are commercially valuable, endangered, species with problematic reproduction, using a more common or easily available species or species adapted to artificial reproduction as a surrogate host. The main goal of this technology is to establish a small-bodied surrogate broodstock producing functional donor gametes based on germ cell transplantation. Extent preliminary experiments, including documentation of donor/host embryonic and larval development, characterization of germ cells enriched by documentation of their migratory activities, sterilization of the host, isolation and cryopreservation of donor germ cells, are key factors for launching this biotechnology. All these crucial points were the main objective of the present work. The whole thesis provided the focus on two different fish species. First, our commercially valuable fish, the tench, where we would like to apply our current knowledge to create a germ-line chimera within cyprinids by transplantation of tench germ cells to smaller and faster-reproducing fish species as white cloud mountain minnow. Secondly we focused on the endangered species (listed in IUCN Red List) of large body size with long reproductive cycle, the sturgeons. In this case, we have chosen sterlet as a host, providing an advantage of shorter generation interval and smaller body size, to produce gametes of donor, a critically endangered species of large body size with long reproductive cycle, such as beluga. This innovative technology could result in collection of sperm and eggs in shorter time from small-bodied host. In tench we firstly focused on embryonic and larval development documentation together with description of origin and migration pathways of primordial germ cells (PGCs). PGCs represent a powerful tool for creation a germ-line chimera within fish species because they transmit genetic information to the next generation (Linhartova et al., 2014a). Secondly, we reported a practical technique for isolation and cryopreservation of early stages of germ cells (GC), including spermatogonia (SG) and spermatocytes (Linhartova et al., 2014b). In case of sturgeons, Saito et al. (2014) firstly described the origin and migration patterns of sturgeon PGCs deposited at the vegetal pole of the egg similar to that in anurans. Secondly, Psenicka et al. (2015) reported isolation and cryopreservation of female and male GC, SG from testes and OG and pre-vitellogenic oocytes from ovary, of 2-4-year old Siberian sturgeon. Moreover the isolated GC were transplanted into host (sterlet) and process of transplantation resulted in successful colonization of sterlet genital ridge. The potential host for germ-cell tranplantation, sterlet, was sterilized by knock-down of germ cell specific gene, the dead end gene, by the morpholino antisense oligonucleotide (MO) agent (dnd-MO). These results reported the first known and functional method of sturgeon sterilization (Linhartova et al., manuscript). We provided important information on morphology and ultrastructure of beluga spermatozoa structure by scanning and transmission electron microscopy to increase knowledge of evolutionary and taxonomic relationships among sturgeons (Linhartova et al., 2013). Finally, this thesis presents several studies with differing focus of research but with one target goal to induce germ-line chimerism in fish. All these results are prerequisite of future application and development of surrogate production in these species.

In this study, for the first time we used CRISPR/Cas9 gene editing technology in sturgeons i.e., sterlets (Acipenser ruthenus). The sequences of sgRNA and primers were designed based on published dnd1 sterlet sequence. Each pair of sgRNA oligos after ligation ready duplex DNA fragment was cloned into vector pX330-U6-Chimeric_BB-CBh-hSpCas9 backbone and thereafter the transformation to competent cells Escherichia coli DH5 was done. The plasmid carried sgRNA was extracted for downstream applications. We diluted extracted plasmid with 10% of 2 M KCl and injection into animal pole of fertilized eggs of sterlets at one to four-cell-stage, 4 hours post fertilization (hpf). At the same time, second microinjection with 2.5% FITC-biotin-dextrans was injected into vegetal pole for labelling PGCs. In the control groups, the eggs were only injected by 2.5% FITC into vegetal pole. PGCs of sterlet were visualized and photographed using a uorescent stereo microscope Leica M165 FC. To confirm the presence or deletion/insertion occurring in the target gene, we used MCE-202 MultiNA microchip electrophoresis system for DNA analysis, in which the targeted gene after amplifying by PCR was analyzed. Mutations in both treated and control embryos of sterlet were further assessed by Sanger sequencing of the PCR product. In present study, we successfully established basic protocols such as preparation of competent cells, construction of vector carrying sgRNA and its transformation into competent cells to carry out the CRISPR/Cas9 technology in sturgeons. Less number of PGCs was observed in embryos that were treated with CRISPR/Cas9; however, sequencing did not provide us a reliable evidence for mutation of the targeted gene probably due to an unspecific PCR. Therefore, more authentication of dnd1 knockout should be done in future by more specific PCR and repeated sequencing.

La presente Tesis Doctoral titulada "Estudio del comportamiento estacionario y dinámico de una pila de combustible de 300 W de potencia operando en modos ánodo cerrado y ánodo abierto" se estudia el efecto de las variables de operación de la pila de combustible como son los caudales de hidrógeno y de aire, la temperatura de operación, la temperatura de humidificación de los gases, los coeficientes estequiométricos de los gases y la presión de alimentación de los mismos sobre el comportamiento de una pila de combustible de 300W y de las celdas individuales que la componen, mediante las curvas potencial-densidad de corriente o curvas de polarización y mediante espectroscopía de impedancias electroquímicas. Para llevar a cabo el trabajo se ha desarrollado un sistema de monitorización y control de las variables mencionadas previamente. Las curvas de polarización son características de cada pila de combustible, y permiten estudiar su comportamiento estacionario en diferentes condiciones de operación. En modo ánodo cerrado el potencial aumenta con el coeficiente estequiométrico del aire. En modo ánodo abierto, el potencial aumenta con los coeficientes estequiométricos del hidrógeno y del aire, con los caudales de ambos gases y con la presión. Con respecto a la temperatura, en ambos modos de operación, los mejores resultados se obtienen cuando la temperatura de operación y de humidificación tienen valores similares. Se ha desarrollado un modelo matemático que permite determinar los parámetros cinéticos de las celdas individuales mediante el ajuste de las curvas de polarización experimentales al modelo propuesto. Al estudiar el comportamiento dinámico se ha observado que cuando se produce un aumento de la densidad de corriente el potencial disminuye bruscamente, alcanza un valor mínimo o subimpulso y luego aumenta hasta alcanzar un nuevo estado estacionario. Por otro lado, cuando se produce un cambio negativo en la densidad de corriente el potencial aumenta hasta un valo / Pérez Page, M. (2012). ESTUDIO DEL COMPORTAMIENTO ESTACIONARIO Y DINÁMICO DE UNA PILA DE COMBUSTIBLE TIPO PEM DE 300 W OPERANDO EN LOS MODOS DE OPERACIÓN DE ÁNODO CERRADO O DEAD-END Y ÁNODO ABIERTO [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/16960 / Palancia

Membrane distillation (MD) is an emerging technology capable of treating high-saline feeds and operating with low-grade heat energy. However, commercial implementation of MD is limited by so-called temperature polarization, which is the deviation in the temperature at the feed-membrane interface with respect to the bulk fluid. This work presents solutions to alleviate temperature polarization in MD by employing a localized heating concept to deliver heat at the vicinity of the feed-membrane interface. This can be realized in multiple ways, including Joule heating, photothermal heating, electromagnetic induction heating, and nanofluid heating. In the first experiment, a Joule heating concept was implemented and tested, and the results showed a 45% increase in permeate flux and a 57% decrease in specific energy consumption. This concept was further improved by implementing a new dead-end MD configuration, which led to a 132% increase in the gained output ratio. In addition, the accumulation of foulants on the membrane surface could be successfully controlled by intermittent flushing of feedwater. Three-dimensional CFD calculations of conjugate heat transfer revealed a more uniform heat transfer and temperature gradient across the membrane due to the increased feedwater temperature over a larger membrane area. In another approach, a photothermal MD concept was used to heat the feed water locally. A 2-D photothermal material, MXene, recently known for its photothermal property, was used to coat commercial MD membranes. The coated membranes were evaluated under one-sun illumination to yield a permeate flux of 0.77 kg.m$^{−2}$h$^{−1}$ with a photothermal efficiency of 65.3% for a feed concentration of 0.36 g.L$^{−1}$. The system can produce around 6 liters of water per day per square meter of membrane. An energy analysis was also performed to compare the efficiency of various energy sources. Considering the sun as a primary energy source, the performance of different heating modes was compared in terms of performance and scale-up opportunities. Overall this work demonstrates that the application of localized heating will enable the scale-up and the use of renewable energy sources to make the MD process more efficient and sustainable. / The illustrative figure was produced by Ana Bigio, scientific illustrator, KAUST.

The diploma thesis deals with restoration of dead-end meander of the Váh river, which is facing problems with fluctuating water level, inappropriate conditions for fish population and insufficient capacity for water withdrawal to adjacent pond. The thesis consists of description of general characteristics of the area and current state of the dead-end meander, followed by the design of modifications within the meander and technical object at the inlet and hydrotechnical calculations concerning the technical object, excavation works and the time of emptying the meander.

No description available.