Dissertations / Theses on the topic 'Sine sweep'

In questa tesi si �e studiato un metodo per virtualizzare ed emulare le distorsioni armoniche non lineari del suono di uno strumento musicale, l'Ocarina di Budrio, tramite algoritmi implementati in ambiente Matlab. Da un punto di vista matematico tali non linearit�à sono state modellate utilizzando lo sviluppo in serie di Volterra diagonale, una sempli�cazione del caso generale valida quando le non linearit�à dipendono dal valore istantaneo dell'ingresso.

Structural Health Monitoring (SHM) techniques are widely used in a number of Non – destructive Evaluation (NDE) applications. There is a need to develop effective techniques for SHM, so that the safety and integrity of the structures can be improved. Two most widely used SHM methods for plates and rods use either the spectrum of the impedances or monitor the propagation of lamb waves. Piezoelectric wafer – active sensors (PWAS) were used for excitation and sensing. In this study, surface response to excitation (SuRE) and Lamb wave propagation was monitored to estimate the integrity of the round objects including the pipes, tubes and cutting tools. SuRE obtained the frequency response by applying sweep sine wave to surface. The envelope of the received signal was used to detect the arrival of lamb waves to the sensor. Both approaches detect the structural defects of the pipes and tubes and the wear of the cutting tool.

Vibration generated from various sources (engine, road surface, tires, exhaust, etc.) should be considered in the design of a car body. These vibrations travel through transfer systems (drivetrain, suspension, body, etc.) to the steering wheel, seats and other areas where it is detected by the passengers of the vehicle. Transmission routes must be studied and efforts made to keep transfer systems from amplifying vibration and to absorb it instead. Since the superior vibration transfer system is the car body, finite element analysis and experimental vibration analysis are performed on car body-in-white. Body vibration analysis entails understanding and improving the body&rsquo
s dynamic characteristics that act as vibration transfer channels. In the previous study, a finite element model has been created for a car body-in-white available in Automotive Laboratory (Mechanical Engineering Department, Middle East Technical University, Ankara) and its natural frequencies and mode shapes have been determined using finite element analysis software. In this study, vibration tests have been performed on actual car body-in-white. Frequency response functions between 34 response locations and force application point have been measured. Using these frequency response functions, natural frequencies and mode shapes of the body-in-white have been determined. Finite element analysis and experimental results have been compared to evaluate the finite element model reliability.

Bone fractures and pathologic conditions like chronic wounds significantly reduce the quality of life for the patients, which is especially dramatic in an elderly population with considerable multi-morbidity and lead to substantial socio-economic costs. To improve the wound healing capacity of these patients, new strategies for the design of novel multi-functional biomaterials are required: they should be able to decrease extensive pathologic tissue degradation and specifically control angiogenesis in damaged vascularized tissues like bone and skin. Glycosaminoglycans (GAGs) like hyaluronan (HA) and chondroitin sulfate (CS) as important extracellular matrix (ECM) components are involved in several biological processes such as matrix remodeling and growth factor signaling, either by directly influencing the cellular response or by interacting with mediator proteins. This could be useful in functionalizing biomaterials, but native sulfated GAGs (sGAGs) show a high batch-to-batch variability and are limited in their availability. Chemically modified HA and CS derivatives with much more defined characteristics regarding their carbohydrate backbone, sulfate group distribution and sulfation degree are favorable to study the structure-function relationship of GAGs in their interaction with mediator proteins and/or cells and this might be used to precisely modulate activity profiles to stimulate wound healing. By combining collagen type I as the main structural protein of the bone and skin ECM with these GAG derivatives, 2.5-dimensional (2.5D) and 3D artificial ECM (aECM) coatings and hydrogels were developed. These biomaterials as well as the respective GAG derivatives alone were compared to native GAGs and used to analyze how the sulfation degree, pattern and carbohydrate backbone of GAGs influence: i) the activity of tissue inhibitor of metalloproteinase-3 (TIMP-3) and vascular endothelial growth factor-A (VEGF-A) as main regulators of ECM remodeling and angiogenesis, ii) the composition and characteristics of the developed 2.5D and 3D aECMs, iii) the enzymatic degradation of collagen-based aECMs and HA/collagen-based hydrogels, iv) the proliferation and functional morphology of endothelial cells. Surface plasmon resonance (SPR) and enzyme linked immunosorbent assay (ELISA) binding studies revealed that sulfated HA (sHA) derivatives interact with TIMP-3 and VEGF-A in a sulfation-dependent manner. sHA showed an enhanced interplay with these proteins compared to native GAGs like heparin (HEP) or CS, suggesting a further impact of the carbohydrate backbone and sulfation pattern. sGAGs alone were weak modulators of the matrix metalloproteinase-1 and -2 (MMP-1 and -2) activity and did not interfere with the inhibitory potential of TIMP-3 against these proteinases during enzyme kinetic analyses. However, the formation of TIMP 3/GAG complexes reduced the binding of TIMP-3 to cluster II and IV of its endocytic receptor low-density lipoprotein receptor-related protein-1 (LRP-1, mediates the up-take and degradation of TIMP-3 from the extracellular environment) in a sulfation- and GAG type-dependent manner. It is of note that the determined complex stabilities of TIMP-3 with cluster II and IV were almost identical indicating for the first time that both clusters contribute to the TIMP-3 binding. Competitive SPR experiments demonstrated that GAG polysaccharides interfere stronger with the TIMP 3/LRP-1 interplay than GAG oligosaccharides. The importance of the position of sulfation is highlighted by the finding that a sHA tetrasaccharide exclusively sulfated at the C6 position of the N-acetylglucosamine residues significantly blocked the receptor binding, while CS and HEP hexasaccharides had no detectable effects. Thus, sHA derivatives as part of biomaterials could be used to sequester and accumulate TIMP 3 in aECMs in a defined manner where sHA-bound TIMP-3 could decrease the matrix breakdown by potentially restoring the MMP/TIMP balance. GAG binding might extend the beneficial presence of TIMP-3 into wounds characterized by excessive pathologic tissue degradation (e.g. chronic wounds, osteoarthritis). Mediator protein interaction studies with sHA coated surfaces showed the simultaneous binding of TIMP-3 and VEGF-A, even though the sHA/VEGF-A interplay was preferred. Moreover, kinetic analysis revealed almost comparable affinities of both proteins for VEGF receptor-2 (VEGFR-2), explaining their competition that mainly regulates the activation of endothelial cells. Additional SPR measurements demonstrated that the binding of sGAGs to TIMP-3 or VEGF-A decreases the binding of the respective mediator protein to VEGFR-2. Likewise, a sulfation-dependent reduction of the binding signal was observed after pre-incubation of a mixture of TIMP-3 and VEGF-A with sGAG poly- and oligosaccharides. The biological consequences of GAGs interfering with VEGF-A/VEGFR-2 and TIMP-3/VEGFR 2 were assessed in vitro using porcine aortic endothelial cells stably transfected with VEGFR 2 (PAE/KDR cells). The presence of sHA both decreased VEGF-A activity and the activity of TIMP-3 to inhibit the VEGF-A-induced VEGFR-2 phosphorylation. The same decreased activities could be observed for the migration of endothelial cells. However, if sHA, TIMP-3 and VEGF-A were present simultaneously, sHA partially restored the TIMP-3-mediated blocking of VEGF-A activity. These findings provide novel insights into the regulatory potential of sHA during endothelial cell activation as an important aspect of angiogenesis, which could be translated into the design of biomaterials to treat abnormal angiogenesis. These sHA-containing materials might control the angiogenic response by modulating the activity of TIMP 3 and VEGF-A. The in vitro fibrillogenesis of collagen type I in the presence of sHA derivatives led to 2.5D collagen-based aECM coatings with stable collagen contents and GAG contents that resemble the organic part of the bone ECM. A burst release of GAGs was observed during the first hour of incubation in buffer with the GAG content remaining almost constant afterwards, implying that the number of GAG-binding sites of collagen restricts the amounts of associated GAGs. Moreover, two differently sulfated HA derivatives could for the first time be incorporated into one multi-GAG aECM as verified via agarose gel electrophoresis and fluorescence measurements. This illustrates the multiple options to modify the aECM composition and thereby potentially their functionality. Atomic force microscopy showed that the presence of sHA derivatives during fibrillogenesis significantly reduced the resulting fibril diameter in a concentration- and sulfation-dependent manner, indicating an interference of the GAGs with the self-assembly of collagen monomers. In line with enzyme kinetic results, none of the GAGs as part of aECMs altered the enzymatic collagen degradation via a bacterial collagenase. Thus aECMs were proven to be biodegradable independent from their composition, which is favorable concerning a potential biomedical usage of the aECMs e.g. as implant coatings. HA/collagen-based hydrogels containing fibrillar collagen embedded into a network of crosslinked HA and sGAGs were developed as 3D aECMs. Scanning electron microscopy demonstrated a porous structure of the gels after lyophilization, which could favor the cultivation of cells. The presence of collagen markedly enhanced the stability of the gels against the enzymatic degradation via hyaluronidase, something beneficial to clinical use as this is often limited by the generally fast breakdown of HA. Binding and release experiments with lysozyme, as positively charged model protein for e.g. pro-inflammatory cytokines, and VEGF A revealed that the sulfation of GAGs increased the protein binding capacity for pure GAG coatings and retarded the protein release from hydrogels compared to hydrogels without sGAGs. Moreover, the additional acrylation of sHA was shown to strongly reduce the interaction with both proteins when the primary hydroxyl groups were targets of acrylation. This stresses the influence of the substitution pattern on the protein binding properties of the GAG derivatives. However, hydrogel characteristics like the elastic modulus remained unaffected. The different interaction profiles of lysozyme and VEGF-A with GAGs demonstrated a protein-specific preference of different monosaccharide compositions, suggesting that the mediator protein binding could be simultaneously adjusted for several proteins by combining different GAG derivatives. This might allow the scavenging of pro-inflammatory cytokines and at the same time a binding and release of wound healing stimulating growth factors. Since there is a growing demand for biomaterials to regenerate injured vascularized tissues like bone and skin, endothelial cells were used to examine the direct effects of solute GAGs and hydrogels containing these GAGs in vitro. In both cases, sHA strongly enhanced the proliferation of PAE/KDR cells. A VEGFR-2-mediated effect of GAGs on endothelial cells as underlying mechanism is unlikely since GAGs alone did not bind to VEGFR-2 and had no influence on VEGFR-2 phosphorylation. Other factors like GAG-induced alterations of cell-matrix interactions and cell signaling could be responsible. In accordance with SPR results, a decreased endothelial cell proliferation stimulating activity of VEGF-A was observed in the presence of solute GAGs or after binding to hydrogels compared to the respective treatment without VEGF-A. However, tube formation could be observed in the presence of solute VEGF A and GAGs and within hydrogels with sGAGs that released sufficient VEGF-A amounts over time. Overall the presence of GAGs and VEGF-A strongly promoted the endothelial cell proliferation compared to the treatment with GAGs or VEGF-A alone. Thus, HA/collagen-based hydrogels functionalized with sHA derivatives offer a promising option for the design of “intelligent” biomaterials that direct and regulate the cellular behavior instead of simply acting as inert filling material. They could be used for the controlled delivery and/or scavenging of multiple mediator proteins, thus enhancing the local availability or reducing the activity of these GAG-interacting mediator proteins, or by directly influencing the cellular response. This might be applied to a range of pathological conditions by tuning the biomaterial compositions to patient-specific needs. However, extensive in vivo validation is required to show whether these in vitro findings could be used to control the biological activity of for instance TIMP-3 and VEGF-A, especially under the pathological conditions of extended matrix degradation and dysregulated angiogenesis.

Oggetto del presente studio riguarda la verifica della conformità acustica per il progetto inerente alla ricostruzione del Teatro Amintore Galli a Rimini, di alcune zone e componenti edilizie, nonché la qualità del campo sonoro nella cavea e nel palcoscenico, con obiettivo prefissato quale il raggiungimento del benessere acustico, intendendo con questa espressione l’esigenza di evitare la presenza di rumori causati da sorgenti interne all'edificio stesso o prodotto da attività in aree circostanti.

In this work a new current-sweep stress methodology for quantitatively assessing the mixed-mode reliability (simultaneous application of high current and high voltage) of advanced SiGe HBTs is presented. This stress methodology allows one to quickly obtain the complete damage spectrum of a given device from a particular technology platform, enabling better understanding of the complex voltage, current, and temperature interdependence associated with electrical stress and burn-in of advanced transistors. We consistently observed three distinct regions of mixed-mode damage in SiGe HBTs, and find that hot carrier induced damage can be introduced into SiGe HBTs under surprisingly modest mixed-mode stress conditions. For more aggressively scaled silicon-germanium technology generations, a larger percentage of hot carriers generated in the collector-base junction are able to travel to and hence damage the EB spacer, leading to enhanced forward-mode base current leakage under stress. A new self-heating induced mixed-mode annealing effect was observed for the first time under fairly high voltage and current stress conditions, and a new damage mechanism was observed under very high voltage and current conditions. Finally, as an example of the utility of our stress methodology, we quantified the composite mixed-mode damage spectrum of a commercial third-generation (200 GHz) generation SiGe HBT. It is found that if devices are stressed with either voltage or current alone during burn-in, they can easily withstand extreme over-stress conditions. Unfortunately, devices were easily damaged when stressed with a combination of stress voltage and current, and this has significant implications for the device and circuit lifetime prediction under realistic mixed-signal operating conditions.

Doctor of Philosophy
Department of Agronomy
Tesfaye Tesso
Sweet Sorghum [Sorghum bicolor (L.) Moench] is a type of cultivated sorghum grown primarily for its sugar-rich stalks. Because of its high fermentable sugar content, the crop is widely recognized as an alternative feedstock source for bio-fuel production. The extent to which stalk sugar accumulation occurs may be determined by several factors including the sink size. Grain is the most important sink in sorghum and other grain crops. Three experiments were conducted in this study to determine the extent to which the grain sink can reduce sugar accumulation in the stalks, to test and validate a genetic system that allows development of sterile sweet sorghum hybrids, and to assess the potential of sugar-rich hybrids to overcome stalk rot diseases. The first experiment, based on 22 sweet sorghum genotypes, was undertaken to study the effect of eliminating the grain sink (removing the head prior to anthesis) on stalk juice yield, sugar accumulation, and biomass. The data showed that the grain sink had a significant effect on all traits measured. Elimination of the grain sink significantly increased oBrix % (17.8%), dry biomass (27.8%), juice yield (23.9%), and total sugar yield (43.5%). The second experiment was aimed at validating the role of A3 genetic male sterility system for producing sterile sweet sorghum hybrids. Ten sweet sorghum pollinator lines of variable sugar content were selected among the entries included in the previous experiment. The lines were crossed to four A1 and A3 cytoplasmic male sterile (CMS) lines using a Design II mating scheme. The A3 females did not have effective restorers so that the hybrids were expected to be sterile. The parental lines and corresponding hybrids were evaluated for biomass production, oBrix, juice and sugar yield using a randomized complete block design. All A3 hybrids were sterile and did not produce seed when heads were covered prior to pollination. The effect of grain sink represented by the A1 vs. A3 CMS were highly significant for Brix%, biomass, juice, and sugar yield. Comparison of parents vs. crosses component was highly significant, indicating marked heterosis effect for the traits. Both general (GCA) and specific (SCA) combining ability effects were also significant for all traits, indicating the role of both additive and dominance genetic effects in the inheritance of the characters. Earlier studies have shown positive relationships between stalk sugar concentration and stalk rot disease resistance in sorghum. Thus, the objective of the third experiment was to study the effects of the CMS mediated differential accumulation of stalk sugar on severity of charcoal rot disease caused by Macrophomina phaseolina. The experiment provided an opportunity to test the effect of variable stalk sugar in the same genetic backgrounds. The data indicated that hybrids produced from A3 cytoplasm were more resistant to charcoal rot (7.1cm lesion length) compared to those produced from the A1 hybrids (9.5 cm lesion length). The enhanced resistance of hybrids with higher sugar yield could have significant agronomic advantage in sugar based bio-fuel feedstock production.

Sex chromosomes are useful for the study of how factors such as mutation, selection, recombination and effective population size affect diversity and divergence.

A comparison of gametologous introns in seven different bird species revealed a complete lack of diversity on the female-specific W chromosome. In contrast, Z had at least one segregating site in all examined species. This can be explained by the lower mutation rate and lower effective population size of W but also suggests that selection affects diversity levels on the non-recombining W chromosome.

In a diverse set of chicken breeds, the Z chromosome showed reduced diversity compared to autosomes and significant heterogeneity in levels of variation. High variance in male reproductive success, leading to a reduced Z chromosome effective population size, can partly explain this observation. In addition, we suggest that selective sweeps frequently act on the Z chromosome and are responsible for a significant part of the observed Z reduction.

Differences in the mutation rate of Z and W chromosome sequences indicate that the time spent in male germ line is important for the mutation rate, but does not exclude a specifically reduced mutation rate on the Z chromosome. Estimates of mutation rate in autosomal, Z- and W-linked chicken and turkey sequences indicate a slight reduction in the rate on Z. However, due to rate heterogeneity among introns this reduction is not significant and we cannot exclude male biased mutation as the single cause of rate variation between the chromosomal classes.

Analysis of indel mutation rates in avian and mammalian gametologous introns show frequent occurrence of indels on both W and Y, excluding meiotic recombination as the only source of this type of mutation. The different indel rate patterns in birds (Z>W) and mammals (X=Y) suggest that indels are caused by both replication and recombination.

Abstract Demographic history and natural selection are central forces shaping the genetic diversity of populations. Knowledge on these forces increases understanding of processes shaping genetic variability of populations. In this PhD thesis I investigated demographic history and selection in multiple populations of Arabidopsis lyrata, an outcrossing herbaceous plant species of the Brassicaceae family. Due to its wide distribution in the temperate and boreal regions, A. lyrata serves as a good model system to study population genetic consequences of colonization of northern latitudes. The first aim of this study was to characterize the demographic and colonization history of the species using site frequency spectra estimated from whole-genome diversity data. Another aim was to detect genetic loci targeted by recent selective sweeps at genome-wide scale as well as at candidate flowering time genes. Patterns of genome-wide selection at linked sites (linked selection) were also compared between populations of Capsella grandiflora and A. lyrata with contrasting demographic histories. Evidence for strong effective population size decline in the past few hundred thousand years was detected in A. lyrata populations species-wide. This study also suggests recent Scandinavian colonization from an unknown refugium, distinct from the Central European source population. Selection analyses revealed loci targeted by positive selection in two Scandinavian lineages after the recent population split as well as selective sweeps in flowering time genes in the colonizing populations. In comparison with the studied C. grandiflora population, the Norwegian A. lyrata population had weaker purifying selection and no evidence for reduction of diversity around genes was found. This thesis offers novel information on species colonization history and its genome-wide effects, which is important for understanding the framework of local adaptation
Tiivistelmä Populaation demografinen historia ja luonnonvalinta ovat keskeisiä populaation perinnöllisen muuntelun muokkaajia. Näiden tekijöiden tutkimus on tärkeää eliöiden sopeutumisen ymmärtämiselle. Tässä väitöskirjassa tutkin demografista historiaa ja valintaa monivuotisen ristisiittoiseen ruohovartisen Brassicaceae-heimon kasvilajin idänpitkäpalon (Arabidopsis lyrata) useissa eri populaatioissa. Idänpitkäpalko on erinomainen mallilaji pohjoiseen ympäristöön sopeutumisen tutkimukseen, koska sen toisistaan eristäytyneet paikalliset populaatiot ovat levittäytyneet laajalle boreaalisella ja lauhkealla ilmastovyöhykkeellä. Tutkimuksen tarkoituksena oli luonnehtia populaatioiden demografista historiaa ja kolonisaatioreittejä käyttäen koko perimän laajuisesta muunteluaineistosta estimoituja alleelifrekvenssispektrejä. Lisäksi koko perimän laajuista aineistoa sekä kukkimisaikaa ohjaavien geenien sekvenssejä käytettiin positiivisen luonnonvalinnan merkkien tunnistukseen. Genominlaajuista kytkeytynyttä valintaa vertailtiin toiseen ristisiittoiseen Brassicaceae-heimon lajin Capsella grandifloran populaatioon, jonka demografinen historia poikkeaa huomattavasti tutkituista idänpitkäpalon populaatioista. Tutkimuksessa havaittiin, että kaikissa tutkituissa idänpitkäpalon populaatioissa tehollinen populaatiokoko oli pienentynyt viimeisen muutaman sadantuhannen vuoden aikana. Kolonisaatiohistorian tarkastelu osoitti, että idänpitkäpalon skandinaaviset populaatiot ovat todennäköisesti peräisin keskieurooppalaisesta refugiosta erillisestä läntisestä refugiosta. Skandinavian kolonisaation yhteydessä vaikuttaneen positiivisen luonnonvalinnan merkkejä havaittiin useissa eri genomin osissa sekä erityisesti valojaksoa mittaavissa geeneissä. Tämä kertoo erilaisiin valojaksoihin sopeutumisen tärkeydestä skandinaavisen kolonisaation yhteydessä. Verrattuna tutkittuun C. grandifloran populaatioon, idänpitkäpalolla puhdistavan valinnan havaittiin olevan heikompaa ja muuntelun vähenemistä geenien ympärillä ei havaittu. Tämä tutkimus tarjoaa uutta tietoa Skandinavian kolonisaatiohistoriasta ja sen genominlaajuisista vaikutuksista. Tutkimuksessa tuotettua tietoa voidaan hyödyntää paikallisen sopeutumisen ymmärtämisessä

Les Héparanes Sulfates (HS) sont de polysaccharides complexes impliqués dans de nombreux processus biologiques. La structure des HS est contrôlée à la surface cellulaire par une famille particulière d‟endosulfatases extracellulaires, les Sulfs. Les Sulfs modifient dramatiquement les propriétés fonctionnelles des HS et sont impliqués dans de nombreux processus physiopathologiques, notamment le cancer. Ces enzymes se composent de deux domaines: un domaine catalytique (CAT) contenant le site actif et un domaine basique hydrophile (HD) responsable de la liaison aux HS. Le but de mon projet de thèse est de caractériser les propriétés structurales et fonctionnelles de la forme humaine HSulf-2, qui demeure à ce jour très mal connues. Dans ce cadre, nous avons tout d‟abord étudié les mécanismes de reconnaissance enzyme/substrat et caractérisé deux nouveaux motifs de reconnaissance des HS sur ces enzymes, responsable de leur activité. En utilisant des oligosaccharides naturels et synthétiques, nous avons aussi démontré que le domaine HD n'est pas essentiel pour la reconnaissance des HS, mais est permet une désulfatation processive et orientée du polysaccharide. De plus, nous avons identifié un tétrasaccharide comme étant la taille oligosaccharidique minimale requise pour l'activité de HSulf-2. Nos résultats nous ont permis de proposer un nouveau modèle décrivant le processus de désulfatation du HS par HSulf-2. D'autre part, nous avons montré que HSulf-2 est un protéoglycane, car il contient une modification post-traductionnelle unique (chaîne CS de Chondoitin Sulfate) sur son domaine HD. Cette chaîne diminue l'activité enzymatique et la liaison aux HS in vitro. Dans le microenvironnement tumoral, en utilisant un modèle de tumeur mammaire orthotopique murin, nous avons montré que la chaîne CS est libérée par protéolyse, conduisant à l'activation de HSulf-2, augmentant la capacité des tumeurs à se développer et à se transformer en métastase. Finalement, nous avons réalisé une étude structurale des Sulfs. Nous avons choisi d‟étudier séparément les deux domaines (CAT et HD). Des essais de cristallogenèse ont été menés pour le domaine CAT afin de résoudre sa structure par cristallographie aux rayons X, mais n‟ont pu aboutir. En ce qui concerne le HD, nous avons mis en place un protocole de production et de purification de HD d‟une manière recombinante et nous avons initiés une étude par RMN ainsi que d'autres techniques biophysiques afin de caractériser structuralement le domaine et d'identifier les sites de liaison aux HS. Nos résultats préliminaires suggèrent que la HD est un domaine non structuré, à l'exception de ses parties N- et C-terminales. L‟ensemble de ces travaux devrait nous permettre de mieux comprendre ces importants mécanismes de régulation des HS et de d‟envisager de nouvelles stratégies anticancéreuses ciblant les Sulfs
Heparan Sulfate (HS) are complex polysaccharides involved in many biological processes. The structure of HS is regulated at the cell surface by unique extracellular endosulfatases, the Sulfs. Sulfs dramatically change HS functional properties, thereby being implicated in many physiopathological processes including cancer. Sulfs features two domains: a catalytic domain (CAT) that comprises the active site, and an hydrophilic basic domain (HD) responsible for HS binding. The aim of my PhD project is to characterize the structural and the functional properties of the human for HSulf-2, which remains poorly understood. In this context, we have first studied the enzyme/substrate recognition mechanisms. We identified two novel HS binding motifs on these enzymes implicated in their activity. In addition, using natural and synthetic oligosaccharides, we demonstrated that the HD is not essential for HS recognition, but is directs the processive and orientated desulfation of the polysaccharide. Moreover, we showed that a tetrasaccharide is the minimal oligosaccharide size required for HSulf-2 activity. Our results enabled us to propose a new model depicting the desulfation process of HS by the Sulfs. Second, we have shown that HSulf-2 is a proteoglycan, given that it harbors a unique PTM (Chondroitin Sulfate, CS chain) on its HD domain. This chain decreases enzyme activity and HS binding in vitro. In the tumoral microenvironment, using a murine orthotropic mammary tumor model, we showed that the CS chain is lost by proteolytic processing, leading to the activation of HSulf-2, and the promotion of tumor growth, vascularization and metastasis. Finally, we have undertaken the structural characterization of the Sulfs. For this, we decided to study separately the two domains found in these enzymes (CAT and HD). Crystallogenesis assays were undertaken for the CAT domain to solve its structure by X-ray crystallography, but were unsuccessful. Regarding the HD, we set up a protocol of production and purification of recombinant HD and we initiated NMR studies and other biophysics analyses in order to structurally characterize the domain and to identify the HS binding sites. Our preliminary results suggest that the HD is an unstructured domain, except for its N- and C-terminal parts. Overall, our data provide significant insights into this critical regulatory step of HS function

FREIRE, R. A. Evaluation of the coarse aggregate influence in the fatigue damage using fine aggregate matrices with different maximum nominal sizes. 2015. 86 f. Dissertação (Mestrado em Engenharia de Transportes) – Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2015.
Submitted by Marlene Sousa (mmarlene@ufc.br) on 2015-10-29T12:42:00Z No. of bitstreams: 1 2015_dis_rafreire.pdf: 6423026 bytes, checksum: 9ed8a164fac500a20720f0a2df391b94 (MD5)
Approved for entry into archive by Marlene Sousa(mmarlene@ufc.br) on 2015-11-03T18:17:39Z (GMT) No. of bitstreams: 1 2015_dis_rafreire.pdf: 6423026 bytes, checksum: 9ed8a164fac500a20720f0a2df391b94 (MD5)
Made available in DSpace on 2015-11-03T18:17:39Z (GMT). No. of bitstreams: 1 2015_dis_rafreire.pdf: 6423026 bytes, checksum: 9ed8a164fac500a20720f0a2df391b94 (MD5) Previous issue date: 2015-07-01
Fatigue cracking is one of the most common distresses in flexible pavements in Brazil and in the world. Fatigue damage in asphalt mixtures originates as a small scale phenomenon. Recently, some authors have used the fine portion of the asphalt mixtures, called Fine Aggregates Matrix (FAM), as an intermediate step to estimate the hot mix asphalt (HMA) behavior. By definition, the FAM is composed of fine aggregates, mineral filler and asphalt binder. There are many volumetric issues of these kind of mixtures. In Brazil, the developed research regarding FAM uses sieve #10 (2.00mm) in the FAM sample preparation, since the #16 sieve (most used in the literature) does not belong to the set of national standardized sieves. Recently, some authors have used the concept of continuum damage mechanics in viscoelastic media in order to quantify the damage during the fatigue testing. In this theory the cracks are considered homogeneous dispersed in the medium, and are represented from internal state variables, determined experimentally. The objective of this study is to evaluate the influence of the variation of one of the FAM constituents, the aggregates, by varying its Maximum Nominal Size (MNS), on damage resistance of these asphalt mixtures. To achieve the objective, Linear Amplitude Sweep (LAS) and time sweep (TS) tests at controlled stress and controlled strain modes of loading were performed on the mixtures and the results were analyzed using the simplified viscoelastic continuum damage (S-VECD) methodology creating C vs S curves. Through the results and a failure criterion based on energy, cyclic tests can be simulated at different loading conditions for Wöhler’s curves construction. The FAM dynamic moduli obtained are very similar to the frequency used to induce damage. However, for lower frequency, the intermediate FAM presented lower stiffness value. It was concluded that FAM with different MNS present very distinct damage performance, and the decrease in the MNS results in an increase in the mixture damage performance for low strain amplitudes. Also, the LAS testing protocol does not allow failure prediction due to the absence of failure in this type of test for FAM.
O Trincamento por fadiga é um dos problemas mais comuns em pavimentos flexíveis no Brasil e no mundo. Danos por fadiga em misturas asfálticas se origina como um fenômeno de pequena escala. Recentemente, alguns autores têm utilizado a porção fina das misturas asfálticas, chamado Matriz de Agregados Finos (MAF), como um passo intermediário para estimar o comportamento do Concreto Asfáltico (CA). Por definição, a MAF é composta de agregados finos, fíler mineral e ligante asfáltico. Há muitas questões volumétricas neste tipo de mistura. No Brasil, a pesquisa desenvolvida sobre MAF usa peneira #10 (2,00mm) na preparação de amostras, uma vez que a peneira #16 (mais utilizado na literatura) não pertence ao conjunto de peneiras normatizadas nacionalmente. Recentemente, alguns autores têm utilizado o conceito de mecânica do dano contínuo em meios viscoelásticos, a fim de quantificar o dano durante o ensaio de fadiga. Nesta teoria as trincas são consideradas homogêneas dispersa no meio, e são representados como variáveis ​internas ​de estado determinadas experimentalmente. O objetivo deste estudo é avaliar a influência da variação de um dos constituintes da MAF, os agregados, variando seu tamanho máxima nominal (TMN), na resistência ao dano destas misturas asfálticas. Para alcançar o objetivo, ensaios de Linear Amplitude Sweep (LAS) e Time Sweep (TS) à tensão controlada e à deformação controlada foram realizadas nas misturas e os resultados foram analisados ​​usando o método simplified viscoelastic continuum damage (S-VECD) para a criação de curvas C vs S. Através dos resultados e um critério de falha com base em energia, os testes cíclicos podem ser simulados em diferentes condições de carregamento para a construção da curva de Wöhler. Os módulos dinâmicos obtidos das MAFs são muito semelhantes aos da frequência utilizada para induzir danos. No entanto, para menor freqüência, a MAF intermediária apresentou menor valor de rigidez. Concluiu-se que as MAFs com diferentes TMNs apresentaram desempenhos de dano muito distintos, e à diminuição do TMN da mistura resulta em um aumento da resistência ao dano para baixas amplitudes de deformação. Além disso, o protocolo de teste LAS não permite a previsão de falha, devido à ausência de falha neste tipo de ensaio para MAF.

Dans l'optique de produire des agro‐carburants, le sorgho sucré est aujourd'hui proposé comme une alternative à d'autres espèces cultivées à grande échelle comme la canne à sucre et le maïs car il présente plusieurs avantages : le sorgho est résistant à la sécheresse et à la chaleur, il nécessite peu d'intrants, a en moyenne un cycle de culture relativement court (3‐4 mois) comparé à la canne à sucre. Il offre une grande diversité génétique à explorer et exploiter, tout en étant génétiquement moins complexe que la canne à sucre. Finalement, il peut être cultivé pour un double usage, le grain pouvant être utilisé comme source d'alimentation pour l'homme ou le bétail (à partir du grain) et le jus sucré contenu par les tiges comme source d'agrocarburant. Cette polyvalence en fait une culture idéale pour lutter contre la compétition entre cultures énergétiques et cultures vivrières et assurer des rendements dans des environnements de culture sujets au stress hydrique et thermique comme c'est le cas en Afrique de l'Ouest. Cependant, le caractère sucré du sorgho est complexe, car sous l'influence d'interactions Génotype X Environnement (GxE). Aussi, les mécanismes métaboliques, morphologiques ou phénologiques constituant la cinétique d'accumulation des glucides dans la tige et son éventuelle compétition avec le remplissage des grains restent mal connus ou très controversés dans la littérature. La présente thèse, réalisée dans le cadre du projet européen Sweetfuel, vise à comprendre ces mécanismes, afin de contribuer à la définition d'idéotypes de sorgho double usage, pour les environnements soudano‐sahéliens.Sur la base d'études expérimentales au champ au Mali et en serre en France, il a pu être démontré que les glucides sont accumulés dans les entrenoeuds des tiges par un jeu d'activités enzymatiques (favorisant l'accumulation d'hexoses puis de saccharose) dès le début de leur élongation, donc potentiellement avant la floraison. Au Mali, l'étude au champ de 14 génotypes adaptés aux conditions locales, plus ou moins sensibles à la photopériode et semés à trois dates différentes, a démontré le bénéfice d'un rallongement de la phase végétative sur la quantité de sucre accumulée dans les tiges de la plante à floraison, du fait d'un plus grand nombre d'entrenoeuds allongés et du temps à leur disposition pour accumuler des glucides avant ce stade. Ce bénéfice était cependant plus lié à la plus grande quantité de biomasse accumulée (taille des tiges) qu'à la concentration en sucre dans les entrenoeuds (plutôt stable entre dates de semis).Ainsi, la durée de la phase végétative et la sensibilité à la photopériode sont proposés comme des paramètres clés favorisant la quantité de glucides accumulée dans les tiges de la plante à floraison. D'autre part, il a été montré que la quantité de glucides présente à maturité dans les tiges des mêmes génotypes ne différait pas ou peu de celle à floraison, une éventuelle réduction pour quelques génotypes n'étant généralement pas significative et évitable par l'allongement du cycle. De plus, cette quantité de glucides dans les tiges à maturité n'a tiré aucun bénéfice de l'ablation de la panicule à floraison chez les mêmes génotypes. Ces résultats suggèrent que la compétition entre le remplissage du grain et la production de sucre est faible chez le sorgho, d'autant plus faible que la plante présente de grandes tiges et donc un grand compartiment de stockage des glucides, tamponnant cette éventuelle compétition. D'ailleurs, à une échelle plus fine, aucune différence n'a pu être mise en évidence en termes d'activité des principales enzymes du métabolisme carboné dans la tige d'un génotype dans sa version stérile (pas de remplissage du grain) et fertile.Ce travail a démontré le potentiel du sorgho pour une double utilisation dans un contexte soudano‐sahélien et la pertinence d'exploiter la diversité génétique de cette espèce pour cette objectif de sélection. Les résultats ob
Sweet sorghum offers many advantages as an alternative to widely cultivated crops such as corn and sugarcane to produce biofuels: it is resistant to water stress, it requires few inputs; it has a shorter growth cycle compared to sugarcane in particular. Sorghum also exhibits a great genetic diversity and is genetically less complex than sugarcane. Finally, sorghum can be cultivated for dual‐purpose uses, using grains for food or feed and sweet juice for biofuel production. Hence, sorghum is a promising option to reduce the competition for land and (water) resource use between food and fuel, in particular in cropping environments with high drought and heat stress frequency, as in West Africa. However, stem sweetness is a complex trait prone to genotype x environment interactions (GxE). The metabolic, morphological and phenological mechanisms involved in the kinetic of stem sugar accumulation and its possible competition with grain filling are largely unknown or controversial in the literature. The present work is part of the European project Sweetfuel and aims at better understanding these mechanisms and contributing to define dual‐purpose sorghum ideotypes for soudano‐sahelian conditions.Based on field and greenhouse experiments respectively in Mali and France, it was found that sugars start accumulating in stem internodes at the onset of their elongation, i.e. potentially soon before the plant flowers. The successive accumulation of hexose and then sucrose in internodes could be dynamically explained by changes in the activity of key enzymes related to sucrose metabolism. In Mali, a field experiment performed on 14 genotypes, contrasted for photoperiod sensitivity and sown at three planting dates, highlighted the interest of increasing vegetative phase duration to increase sugar yield. This was explained first of all by the higher number of internodes that could expand during a longer vegetative phase, and thus, by the higher production of stem biomass, and, to a minor extent, by the longer time for internodes to mature and accumulate sugar (sugar concentration in the stem was however fairly stable across sowing dates). Also, vegetative phase duration and photoperiod sensitivity can be considered as two key parameters promoting stem sugar content before grain filling. In the same time, it was shown that stem sugar content kept remarkably constant between anthesis and maturity in most of studied genotypes and that the reduction observed for some genotypes was overcome with an early sowing. Moreover, sugar accumulation in the stem between flowering and maturity did not benefit from panicle pruning. These results together suggest that the competition for carbohydrates between stem sugar reserves and grain filling is weak; it is even weaker for big/large stem genotypes with huge sugar reserves in the stem that would buffer a post‐flowering allocation of sugar from the stem to the grains if required. This low competition was confirmed at a finer scale, as no differences were observed in the activity of key enzymes of sucrose metabolism between the sterile and the fertile line of a same genotype.This work demonstrates the potential of sorghum for dual‐purpose in particular for soudano‐sahelian cropping conditions and the interest of using its genetic diversity for this breeding purpose. It provides further knowledge for revisiting the phenotyping strategies to be adopted to investigate the genetic basis of sugar and grain production and their combination. The results are also currently used to improve the way the source‐sink relationships underlying this dual production are formalized in crop and plant models at CIRAD. Such models will be then useful to assist sorghum ideotype exploration for dual purpose

Accelerated durability testing of automotive components has become a major interest as it may predict the life characteristics of the vehicle by testing fatigue failure at higher stress level within a shorter period of time. In this work, a specially designed sub-scaled experimental testing bed with the rigid and elastic supports of a simply supported beam was designed and built to compare the effects of the elastic foundation on the change of modal parameters of the tested structure which was later used to tune the FE model. Afterwards, the accelerated loading profiles of both sine sweep and random vibration were applied on the FE model to compare the deviation of the cumulative fatigue damage between the elastic and rigid supports. This work reveals a significant amount of inaccuracy in the current laboratory testing system where the dynamic properties of the tested structure are not maintained close to the real situation.
October 2016

博士
國立臺灣大學
植物學研究所
87
In an effort to study regulation of starch synthesis in the source and sink tissues, a cDNA clone for starch granule-bound starch synthase (GBSSI), which encodes a 67-kDa protein, was isolated from a cDNA library prepared from tuberous roots of sweet potato (Ipomoea batatas Lam.). This GBSSI gene was well expressed in tuberous roots, leaves, and stems, but not in roots. However, mechanisms involved in regulating the expression of this gene were different between tuberous roots and leaves. In tuberous roots, the synthesis of GBSSI transcript increased coordinately with tuberous root expansion; nevertheless, accumulation rates of GBSSI protein in starch granules remained constant regardless of tuberous root sizes, suggesting an involvement of post-transcriptional regulation for the synthesis of this protein. The levels of GBSSI transcript were also investigated in photosynthetic tissues during diurnal cycles, and the results suggest that the transcription of the GBSSI gene in leaves is controlled by endogenous circadian rhythm. The circadian rhythm of GBSSI transcripts could be maintained for at least 3 cycles even under continuous dark (DD) conditions, but the amplitude decreased gradually. Fluctuations of GBSSI protein accumulation patterns were similar to that of mRNA; however, the peak of protein level was broader than that of mRNA. The data obtained from nuclear run-on transcriptional assays indicated that the biological clock appeared to affect GBSSI gene expression at the level of transcriptional rate. Starch accumulations in the leaf tissue also showed a pattern characteristic of circadian regulation. Supplements of exogenous sucrose into medium could increase GBSSI transcript accumulations but did not change the pattern of circadian rhythm. In conclusions, the GBSSI gene expression in leaves appears to be regulated by two independent pathways: (1) biological clock(s) control the circadian expression of GBSSI gene; (2) light plays as an indirect signal to enhance GBSSI mRNA accumulations mediated by photosynthetic product, sucrose.

碩士
國立雲林科技大學
漢學資料整理研究所碩士班
101
Literature is a reflection of society and one’s experience. Besides , the phenomena of life in the story often deeply disclose a more specific message than in the reality of life. Hai-yin Lin’s “Memories of Peking: South Side Stories” describes the author''s own story of growth and migration experience in Beijing. She wrote down the adult world through the eyes of a little girl. From each main character of the story, she learned the meaning of life. Chi-Chun’s “Sweet Osmanthus Flowers Falling Like Rain Drops” is also a childhood writing. This autobiographical memoirs writes about her experience of migration to Taiwan, and how she deeply missed the passing time and space, in efforts to capture her memories of people, events, and objects. In this paper, the analysis is focus on the nostalgic writing in the “Memories of Peking: South Side Stories” and “ Sweet Osmanthus Flowers Falling Like Rain Drops”. By showing the similarities and differences in form and content of the two, we will present their literary value. The first chapter discusses the life and background of the authors, by which we will understand how they live, learn and grow up in the society of the old and new era alternately. Then by the concept of "nostalgia" theme, we analysis the historical background , space geographical, events, items and characters for every aspect, in order to know Hai-yin Lin’s and Chi-Chun’s creative themes, thoughts and feelings, style and brushwork. Lin used an extremely neat style of writing recounting the life of the former Beijing. On the other hand, Chi-Chun wrote about her feelings and mood out of sorrow and calmness. This two articles both recalled time and space to achieve the purpose of authors’ emotional expression. Meanwhile, their writing themes deeply rooted in the Chinese culture and show its effects in their articles everywhere. This unique writing style not only opened a new page of nostalgic writing, but also expanded the new and old hometown nostalgia literature.