Neben den omnipotenten embryonalen Stammzellen existieren im menschlichen Körper adulte mesenchymale Stammzellen (MSZ). Diese Zellen sind in mesenchymalen Geweben über den gesamten Organismus verteilt und sorgen für die Entwicklung und Erneuerung von mesenchymalen Geweben wie Knochen, Knorpel und Bändern. Daher gelten die MSZ im Gegensatz zu den omnipotenten embryonalen Stammzellen als multipotent. Diese verschiedenen MSZ stellen keine homogene Population dar, zeigen aber sowohl in vivo und auch in vitro ein ähnliches Differenzierungsverhalten. In der vorliegenden Arbeit wurde nun eine aus den Knochentrabekeln selbst isolierte MSZ-Population, so genannte bhMSZ, mit MSZ aus dem Knochenmark, mhMSZ genannt, mittels Array-Analyse miteinander verglichen. Die technische Evaluation des Array respektive der zugehörigen SAM-Analyse (significance analysis of microarrays) mittels konventioneller oder Real-Time PCR diente dazu, die Verlässlichkeit der Aussage der Hybridisierungsverfahren zu überprüfen. Dies wurde mit einem Set an ausgewählten Genen durchgeführt, die signifikant differentiell exprimiert waren, und die im Rahmen der Stammzellbiologie relevant erschienen. Die Analyse zeigte, dass die Übereinstimmung der Aussage im Array in über 80 % mit den Ergebnissen der RT-PCR kongruent war. Auf Grund starker interindividueller Schwankungen zeigte sich aber auch, dass die Anzahl der Spender 5 nicht unterschreiten sollte. Im Rahmen der Untersuchungen ergab sich, dass offenbar bei MSZ der Passage 0 eine Kontamination der MSZ mit Plasmazellen vorliegt. Weitere Versuche zeigten, dass erst das Passagieren der MSZ kontaminierende Plasmazellen weitgehend aus der Zellkultur entfernte. Aus diesem Grund wurde in einer weiteren Array Analyse das Transkriptom von MSZ aus Knochentrabekeln mit MSZ aus dem Knochenmark in Passage 1 verglichen. Es zeigten sich in einer stringenten SAM-Analyse keine Unterschiede im Transkriptom. Für klinische Anwendungen scheinen die bhMSZ daher auf Grund der aufwendigeren Isolierung und des dennoch eher geringen Zellgewinns nicht im gleichen Maß für klinische Anwendungen geeignet wie mhMSZ. / The human body contains besides omnipotent embryonal stem cells also adult mesenchymal stem cells (MSC). These cells are spread all over the organism in mesenchymal tissues and are responsible for the development and regeneration of mesenchymal tissues like bone, cartilage and ligaments. Therefore these cells are called multipotent in comparison to the omnipotent embryonal stem cells. The different types of MSC are not a homogeneous population, but show in vivo and in vitro a similar differentiation behavior. The aim of this work was to compare the population of MSC isolated from bone marrow stroma (bhMSC) with MSC isolated from bone fragments (bhMSC) via microarray analysis. The technical evaluation of this array was performed by conventional or real-time PCR to evaluate the reliability of the hybridization procedure. This was done with an assortment of genes, which were differentially expressed, and were estimated to be relevant for the biology of stem cells. The analysis showed 80% accordance between the results of the microarray analysis and the PCR. In addition, the results showed that the number of the analysed individuals shouldn’t be under 5, because of a high interindividual variability. Besides this the experiments showed, that MSC of passage 0 are obviously contaminated with plasma cells. Further tests showed that contaminating plasma cells were widely removed in cell cultures of MSC passage 1. Therefore an additional array analysis was performed comparing MSC isolated from bone marrow stroma in passage 1 with those isolated from bone fragments. No difference could be observed in terms of their transcriptomes obtained by stringent SAM analysis. For clinical purposes bhMSC seem to be less suited because of the more sophisticated isolation procedure and nevertheless the lower cell number yield.

Untersuchungen der Transkriptionsebene individueller präimplantatorischer Embryonalstadien können wertvolle Informationen über den physiologischen Status der betrachteten Embryonen, die z.B. zur Verbesserung der Systeme zur In vitro-Produktion von Embryonen genutzt werden können, liefern. Bisher fehlte es jedoch an einer geeigneten Technologie, um eine große Anzahl von Transkripten in einzelnen Embryonen zu erfassen. Zielsetzung der vorliegenden Arbeit war es, ein Verfahren zur globalen Amplifikation embryonaler mRNA-Präparationen zu entwickeln, das die Analyse der Transkriptionsebene einzelner präimplantatorischer Embryonalstadien über die cDNA-Array-Technologie ermöglicht. Dazu wurde die Strategie gewählt, zwei bereits etablierte Amplifikationsverfahren, Polymerasekettenreaktion und In vitro-Transkription, zu kombinieren, um so synergistische Effekte beider Verfahren zu nutzen. Die Evaluierung des entwickelten Verfahrens zeigte eine hohe Reproduzierbarkeit der erhaltenen Genexpressionsdaten und belegte, dass die relativen Mengenverhältnisse einzelner mRNA-Spezies zueinander während der globalen mRNA-Amplifikation nur unwesentlich verändert wurden. Die entwickelte Methodik ist somit geeignet, komplexe Genexpressionsprofile einzelner Blastozysten zu erstellen und Unterschiede in der Expressionsstärke einzelner Transkripte zu detektieren. Es konnte weiterhin gezeigt werden, dass es möglich ist, über heterologe Hybridisierung Genexpressionsprofile boviner Blastozysten mit cDNA-Arrays, die murine Probensequenzen enthalten, reproduzierbar darzustellen. Neben der Detektion individueller Unterschiede in den Genexpressionsprofilen diverser muriner Embryonalstadien und boviner Blastozysten lag ein Schwerpunkt dieser Arbeit in der Untersuchung der Auswirkungen verschiedener in vitro-Produktionssysteme auf die embryonale Genexpression. Die erhaltenen cDNA-Array Expressionsdaten muriner Oozyten, Zweizeller und Blastozysten befanden sich dabei in Übereinstimmung mit Daten früherer Publikationen anderer Arbeitsgruppen. Genexpressionsprofile in vitro fertilisierter boviner Blastozysten ließen eine Beurteilung der Auswirkungen unterschiedlicher Proteinsupplemente des Kulturmediums auf die embryonale Genexpression zu. Im Rahmen dieser Arbeit wurden zum ersten Mal Genexpressionsprofile einzelner präimplantatorischer Säugerembryonen über cDNA-Array-Analyse erstellt. Die entwickelte Technologie ermöglicht es -bei Verwendung entsprechender cDNA-Array-Systeme-, eine theoretisch unbegrenzte Zahl von Transkripten in individuellen Säugerembryonen semiquantitativ zu erfassen. Dies ist ein wichtiger Schritt hin zu einem besseren Verständnis komplexer Regulationsabläufe während der frühen Embryonalentwicklung und einer besseren Beurteilung der Lebensfähigkeit und Entwicklungskompetenz in vitro produzierter Embryonen, was für die Verbesserung von In vitro-Produktionssystemen für Embryonen sowohl bei Tieren als auch beim Menschen unerlässlich ist. / Transcript expression profiling in single mammalian embryos can provide valuable information about their physiological status and developmental competence that can be exploited to improve systems for embryo in vitro production. Conventional methodologies such as RT-PCR limit the number of transcripts that can be quantitatively screened in a single embryo to only a few. The purpose of this study was to develop and evaluate a methodology for the global amplification of mRNA that permits cDNA-array analysis of individual preimplantation embryos. For this purpose, two conventional amplification procedures – polymerase chain reaction and in vitro transcription – were combined to a global amplification procedure. Evaluation of methodology developed revealed that data produced were high reproducible and that the relative transcript levels found in the original (non-amplified) sample were maintained throughout the amplification process. Thus, this method is suitable to generate complex gene expression profiles and to detect differentially expressed transcripts in individual mammalian embryos. Furthermore, this study demonstrates that expression profiles can reproducibly be produced from bovine embryos using arrays consisting of murine cDNA-probes by heterologous hybridization. The focus of this study was to establish a methodology to detect differentially expressed genes in different murine developmental stages and in bovine embryos derived from different in vitro production systems. The data obtained from murine oocyte, 2-cell stage and blastocyst expression profiles were in agreement with data previously published by other groups. Expression profiles from bovine in vitro fertilized embryos cultured in different media revealed effects of different media protein supplementation on embryonic gene expression. In this study, for the first time, gene expression profiles were generated from single mammalian preimplantation embryos via model cDNA-arrays. Using state-of-the-art cDNA-arrays this technology features the quantitative screening of a virtually unlimited number of transcripts in individual blastocysts and cleavage stages. Complex expression profiles of preimplantation embryos will contribute to the understanding of the molecular mechanisms essential for embryogenesis. This is crucial for the improvement of systems for in vitro production of mammalian embryos.

An ever-increasing demand for wider bandwidths in communication, radar, and imaging systems has emerged. In order to facilitate this growing demand, progressive research into millimeter-wave technologies has become vital in achieving next generation networks such as 5G. Being cost effective and easy to manufacture, Substrate Integrated Waveguide (SIW) circuits have been demonstrated as a viable candidate for high-frequency applications due to their low-loss, high quality-factor, and high power-handling capabilities.Research on beam-forming networks, specifically the Butler matrix, has demonstrated powerful beam-steering capabilities through the use of passive component networks. Through these clever configurations, a cost effective and robust option is available for us to use. In order to further millimeter-wave research in this area, this thesis presents a modified configuration of the Butler Matrix in SIW that is physically reconfigurable; by separating the Butler matrix from the antenna array at a pre-selected point, the array can be easily interchanged with other 1-Dimensional,and 2-Dimensional slot antenna arrays. Although this system does not fall under the rigorous definitions of Reconfigurable Antennas, it should be noted that the interchangeability of 1 and 2 dimensional arrays is not typically expressed in Butler matrix configurations. Design and simulations are carried out in CST Microwave Studio to inspect individual components as well as system characteristics. Circuit prototypes are then manufactured and tested in an anechoic chamber to validate simulation results and the design approach. / Graduate / 2020-07-17

Student Number : 0000713F - PhD thesis - School of Physics - Faculty of Science / The Doppler shift and the Doppler broadening of prompt gamma emissions were measured for some residues formed in the interaction of 33 A MeV 12C ions with a 63Cu target using the AFRODITE detector array at Faure, Cape Town. This is a potentially new technique to carry out nuclear interaction studies. Coincident gamma rays emitted by the residues are used in their identification. Detection at angles other than 90◦ with respect to the beam axis gives the magnitude of the mean Doppler shifts and the average linear momentum transfer. The Doppler broadening of the detected gamma lines at 90 ◦ with respect to the beam axis could give the residue recoil angular distribution. The precise shapes of the Doppler shifted and broadened gamma lines for each of the residues extracted, reveals the distribution, in magnitude and angle, of the momentum transferred in the interaction process. In addition, characteristic gamma energy transitions of each residue populated carry additional information on angular momentum (spin) transfer, production cross-section and nuclear excitation states. The measured residues show a unique distribution of momentum ranging from single nucleon transfer to complete damping of the projectile momentum. The measured observables are consistent with the existing data from other techniques, making the new technique viable option for studying nuclear interaction kinematics. A comparison of the experimental measurements with the predictions of the model developed in Milano 1 and GEANT4 calculations shows that the model developed in Milano model give a much better agreement compared to the GEANT4 calculations, attributed to the assumption of projectile break-up and re-emission process of some of the fragments during the first step of the nuclear interaction process.

A obesidade se tornou uma das maiores preocupações de saúde pública. É um distúrbio neuroendócrino, no qual fatores ambientais e genéticos agem em conjunto, levando ao excesso de armazenamento de energia na forma de gordura corporal. A síndrome de Prader-Willi (PWS) é a mais freqüente das síndromes que possui a obesidade como uma de suas características, com incidência de 1:25.000 nascimentos. É caracterizada por hipotonia neonatal com dificuldade de sucção, atraso do desenvolvimento neuropsicomotor (DNPM), hiperfagia, obesidade, baixa estatura em adolescentes, mãos e pés pequenos, hipogonadismo, distúrbios do sono, características faciais dismórficas, deficiência intelectual leve a moderada e comportamento obsessivo-compulsivo. Pacientes com atraso do DNPM e/ou dificuldade de aprendizado, distúrbios de comportamento, obesidade e/ou hiperfagia, com teste negativo para PWS, foram estudados com plataformas de SNP array, “The GeneChip® Mapping 500K Set” da Affymetrix, ou array-CGH, CytoSure ISCA 4x180k da OGT, para identificar genes relacionados a obesidade e hiperfagia, assim como, novas regiões genômicas implicadas na etiologia de síndromes genéticas associadas à obesidade. Dentre os 31 pacientes estudados, oito apresentaram variações de número de cópias (CNVs) em seu genoma: deleção em 1p22.1p21.2; deleção em 3q25.33q26.1 e deleção em 13q31.2q32.1; duplicação em 7q36.2; deleção em 8p23.3p23.1 e duplicação em 12p13.33p13.31; duplicação 16p13.11p12.3; duplicação em 17q11.2; deleção em 20p12.1; duplicação em 21q22.13. Duas dessas alterações foram herdadas de pais fenotipicamente normais. Algumas dessas CNVs sobrepõem regiões genômicas previamente relacionadas com obesidade, incluindo a microdeleção de 1p21.3 e as duplicações dos cromossomos 12 e 21. Identificamos genes anteriormente descritos como associados à obesidade (PTBP2, DPYD, MIR137, GNB3 e PPM1L), ou possivelmente envolvidos com este fenótipo (HTR5A e KCNJ6), mapeados em várias dessas CNVs. Além disso, os genes RNF135, NF1, DPP6, GPC5, DYRK1A e MACROD2 são os prováveis causadores da deficiência intelectual, atraso do desenvolvimento neuropsicomotor, dificuldades de aprendizagem, distúrbios de comportamento e outras características clínicas encontrados nos pacientes. O diagnóstico e prognóstico dos pacientes e o Aconselhamento Genético aos pais e familiares é fornecido / Obesity has become a major concern for public health. It is a neuroendocrine disorder, in which genetic and environmental factors act together, leading to excessive storage of energy as fat. Prader-Willi syndrome (PWS) is the main obesity-related syndrome with a birth incidence of 1:25,000. It is characterized by neonatal hypotonia, poor sucking, developmental delay, hyperphagia, obesity, short stature in adolescents, small hands and feet, hypogonadism, sleep disturbance, dysmorphic facial features, mild to moderate intellectual disability and obsessive-compulsive behavior. Patients with psychomotor developmental delay and/or learning disabilities, behavior disorders, obesity and/or hyperphagia, who tested negative for PWS, were studied by chromosomal microarray analysis, including the SNP-based platform “The GeneChip® Mapping 500K Set” (Affymetrix), and the array-CGH platform “CytoSure ISCA 4x180k (OGT)”, to identify genes related to hyperphagia and obesity, as well as new genomic regions implicated in the etiology of genetic syndromes associated with obesity. Of 31 patients studied, eight had copy number variants (CNVs) in the genome: 1p22.1p21.2 deletion; 3q25.33q26.1 deletion and 13q31.2q32.1 deletion; 7q36.2 duplication; 8p23.3p23.1 deletion and 12p13.33p13.31 duplication; 16p13.11p12.3 duplication; 17q11.2 duplicaton; 20p12.1 deletion; 21q22.13 duplication. Two of these CNVs were inherited from an unaffected father. Some of these CNVs overlap genomic regions that have previously been related to obesity, including the 1p21.3 microdeletion and the duplications of chromosomes 12 and 21. Furthermore, we identified genes previously described as associated with obesity (PTBP2, DPYD, MIR137, GNB3 and PPM1L), or possibly involved with this phenotype (HTR5A and KCNJ6), mapped to several of these CNVs. In addition, the genes RNF135, NF1, DPP6, GPC5, DYRK1A and MACROD2 are likely implicated in intellectual disability, developmental delay, learning disabilities, behavioral disorders and other clinical features found in patients. The diagnosis and prognosis of patients and genetic counseling to parents and families is provided

by C.H. Fung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1989. / Bibliography: leaves [81-82]

A new method for the simulation of radio frequency (RF) coils has been developed. This method utilizes the FEM simulation package Ansoft HFSS as a base for the modeling of RF coils with complex biological loading effects. The abilities of this software have been augmented with custom MATLAB code to enable the fast prediction of lumped element values needed to properly tune and match the coil structure as well as to perform the necessary post processing of simulation data in order to quickly generate and evaluate field data of the resonating coil and compare design variations. This method was evaluated for accuracy and implemented in the re-design of an existing four channel breast coil array for clinical imaging of the female breasts. Based on the simulation results, a commercially viable printed circuit board (PCB) implementation was developed and tested in a clinical 1.5 T MR scanner. The new design allows for wide open bilateral access to the breast regions in order to accommodate various interventional procedures. The layout has also increased axillary B1 field coverage with minor penalty to the signal-to-noise ratio of the coil array, enabling high-resolution imaging over a wide field-of-view.

In this thesis the focus was on the design, fabrication, and tests of the feeding networks individually and within an array system. The array feeding network is a corporate-fed type utilizing equal-split, stepped-multiple sections of the conventional Wilkinson power divider in microstrip form with a unique topology. The feeding network was specifically designed for a broadside relatively small linearly-polarized wideband UHF non-scanning array for directed power applications that uses an array radiator with a new volumetric ribcage dipole configuration. The array has a large impedance bandwidth and consistent front lobe gain over the wide frequency band. Theoretical and experimental results describing the performance of the array feeding network and the array are presented and discussed.

Thesis (M.A.)--Boston University / The primary objective of this research is to test and validate a geometric-optical bidirectional reflectance canopy model developed by Li and Strahler, with respect to actual forest canopy reflectance measurments. This model treats forest canopies as scenes of discrete, three dimensional objects that are illuminated and viewed from different positions in the hemisphere. The shapes of the objects, their count densities and patterns of placement are the driving variables, and they condition the mixture of sunlit and shaded objects and background that is observed from a particular viewing direction, given a direction of illumination. This mixture, in turn, controls the brightness apparent to an observer or a radiometric instrument. The Advanced Solid-State Array Spectroradiometer (ASAS) is chosen to be the sensor having the ability of collecting measurements at various look angles and its imaged reflectance was used to validate the model. The modelled BRF's were compared to actual ASAS measured BRF's in sites with different canopy structures and densities. The comparision revealed execellent match between the modelled and measured reflectance, and great ability of the model in predicting the shape and magnitude of the BRDF, in almost all the sites investigated. It is concluded that the geometric optics approach provided a good way to model the bidirectional reflectance distribution function of natural vegetation canopies, that captures the most important features exhibited by bidirectional measurements of such canopies. Further modifications have been suggested that will improve the predicted BRF's, and yield better results. [TRUNCATED]

A novel approach to ionospheric imaging with the purpose of weather/distaster prediction and climate study is introduced. This feasibility study combines traditional material imaging techniques with high frequency (HF) radio via SDR (software defined radio) systems in order to capture three-dimensional images of the atmosphere. An experiment is devised and the necessary instrumentation built in order to capture coherent images of the ionosphere. The experimental results show these three-dimensional images as well as a novel approach to measuring ionospheric height. The novelty of the research comes from the use of a closely spaced phased-array of radio antennas in conjunction with a post-correlation beamformer repurposed from radio astronomy. Experiments were run at both the University of Victoria and DRAO (Dominion Radio Astrophysical Observatory), the results which led to a successful proposal for extending the research onto a larger array with support from research groups in New Mexico. / Graduate