Glioblastoma multiforme (GBM) is the most aggressive and most common primary brain tumor in adults accounting for 50-60% of primary brain tumors. The prognosis for patients with GBM remains poor and treatment is mainly palliative with a mean survival time of less than one year. Radiotherapy is used extensively in the management of glioblastoma either alone or in combination with surgery and/or chemotherapy. However, this tumor is one of the most resistant tumors to radiotherapy thus limiting the benefit of this form of treatment. <p>Studies have shown that malignant tumors have a high content of glutathione an antioxidant responsible for protecting the cells against damage from free radicals (mainly superoxide, hydroxyl and hydrogen peroxide). It is well established that glutathione, by neutralizing these free radicals plays a major role in radioresistance. Glioblastoma has relatively high levels of glutathione. In this study, by reducing the glutathione content of glioblastoma in a rat model, we were able to investigate the effect of this reduction in enhancing the effect of radiotherapy as a form of treatment for glioblastoma multiforme in a rat model. <p>By injecting L-Buthionine-SR-Sulfoximine (BSO) in to the tumor tissue, the glutathione content of the tumor was reduced by about 70% of its initial value. When administered into the tumors 2 hours prior to radiotherapy the animals so treated had a significantly longer median survival time compared with animals that received radiotherapy alone.

During the maturation of pre-mRNA introns are removed and exons are spliced together, to form a primary transcript, a reaction that is catalyzed by the spliceosome. Alternative splicing is a complex reaction that mainly utilizes one of four mechanisms; exon skipping, 5’ splice site choice, 3’ splice site choice and intron retention. To achieve accurate splicing four sequence elements are essential, two of which are located in the splice sites themselves; 5’ splice sites and 3’ splice sites, but also the polypyrimidine tract and the branch point sequence. Alternative splicing can be regulated by histone or chromatin modulations, siRNA, transcription efficiency and various proteins, many of which belong to either the SR protein family or the hnRNP family of proteins. SR proteins usually promote exon inclusion, while hnRNP proteins usually promote exon skipping. There are also regulatory elements that are called exonic splicing enhancers or silencers depending on if they promote or inhibit the inclusion of the exon they reside in. These elements also exist in introns and are then called intronic splicing enhancers or silencers. The enhancer elements are most commonly targeted by SR proteins and the silencer elements are usually targeted by hnRNP proteins. This paper will mainly focus on the regulation of alternative splicing and the role of alternative splicing under abnormal conditions, such as when mutations cause disease.

Alternativ splicing är en process som ger upphov till att olika mRNA-sekvenser bildas från en enda gen, vilket bidrar till en ökad proteindiversitet hos organismen. Olika mRNA-sekvenser kan uppstå eftersom att det förekommer olika varianter av alternativ splicing som även kan kombineras på flera olika sätt: cassette exon (inkludering/exkludering av exon), intron retention (intronet behålls), alternative 5´splice-site choice (olika 5´ splice sites kan väljas) och slutligen alternative 3´ splice-site choice (andra 3´ splice sites kan väljas). För att alternativ splicing ska äga rum i olika pre-mRNA måste den regleras av cis-reglerande element. De cis-reglerande elementen utgörs av fyra grupper: exonic splicing enhancers (ESE), exonic splicing silencers (ESS), intronic splicing enhancers (ISE) samt intronic splicing silencers (ISS). Som namnen förtäljer finns de antingen i exoner eller introner, där de interagerar med transagerande faktorer, SR-proteiner (aktiverare) eller hnRNPs (hämmare). Alternativ splicing förekommer både i djur och i växter. Hos Homo sapiens genomgår över 74 % av de 25,000 gener som finns hos organismen, alternativ splicing. Däremot i växten Arabidopsis thaliana, genomgår endast 22 %, av den totala mängden på cirka 26,000 gener, alternativ splicing. Eftersom att processen bidrar till en ökad proteindiversitet, kommer det medföra att olika processer i organismerna påverkas, exempelvis celltillväxt, celldöd samt utvecklingen av olika sjukdomar, såsom Parkinson och cystisk fibros. Många studier har gjorts som bekräftar dess betydelse för organismerna men på grund av processens komplexitet är det fortfarande ett ämne som ständigt måste utforskas.

<p>SR proteins are a family of splicing factors involved in the regulation of both constitutive and alternative splicing of pre-mRNAs. Despite years of studies, several big questions still remain: how the expression levels of SR proteins are regulated; what are the underlying mechanisms responsible for SR proteins-mediated gene regulation; what are the physiological targets of SR proteins in vivo. In my dissertation study, I am focusing on two members of the family, SF2/ASF and SRp20, to study their functional involvement in regulating microRNA/mRNA biogenesis and their own expression. </p><p>Negative feedback regulation is a common mechanism maintaining the steady-state level of SR proteins (i.e. SC35 and SRp20), and several mechanism may be involved. In order to test if miRNAs are also involved in such negative feedbacks, small RNA sequencing was used to identify differentially expressed miRNAs after SF2/ASF overexpression in an inducible stable cell line system. Among the 40 differentially expressed miRNAs, miR-7 is particularly interesting, because it is also predicted to target SF2/ASF, which forms a negative feedback regulation. This is indeed the case as shown by luciferase reporter assay and overexpression/knocking down of miR-7 in vivo. To our knowledge, this is the first identified negative feedback circuit between a SR protein and a miRNA, which may be a general mechanism in regulating SR protein homeostasis.</p><p>To characterize the mechanism underlying SF2/ASF-enhanced miRNA biogenesis, I have employed a series of molecular and biochemical approaches to pinpoint the key molecular interactions in a minigene system, which is consist of miR-7 embedded intron and the flanking exons of its host gene. By manipulating the splicing pattern of such minigene, I have uncovered a splicing-independent function of SF2/ASF in regulating miRNA biogenesis. Directly binding between SF2/ASF protein and pri-miR-7 was demonstrated by Cross-linking and immunoprecipitation assay (CLIP) and RNA affinity purification. The precise binding site was then pinpointed by combining computational prediction and mutagenesis assay. Finally, by using in vitro pri-miRNA processing assay, I showed that SF2/ASF can promote the Drosha cleavage step of pri-miR-7 through directly association with the predicted binding site. So far, this is the first SR protein discovered, which is directly involved in miRNA biogenesis. Moreover, our preliminary data also suggested that SF2/ASF may promote miRNA biogenesis in other steps after Drosha cleavage; and different SR proteins can regulate miRNA biogenesis in a substrate-specific manner. Taken together, SR family of splicing factors may be broadly involved in miRNA biogenesis through direct interactions.</p><p>In order to study the general involvement of SR proteins in RNA biogenesis, one important step stone is to have a better profile of their targets in vivo. To achieve this, I focused on SRp20, another classic SR protein. Photoactivatable-Ribonucleoside-Enhanced Cross-linking and immunoprecipitation assay combined with deep sequencing (PAR-CLIP-seq) was used to identify the binding partners of SRp20 globally, which is subsidized by candidate gene validations. Consistent with the literature, I found that SRp20 primarily targets exonic regions for splicing regulation, and such interactions are likely to be sequence dependent on the CWWCW motif. Surprisingly, I also observed extensive binding between SRp20 and the 3' UTRs of mRNA, which may affect the choice of alternative polyadenylation sites. The underlying mechanisms are being investigated by a variety of molecular methods. </p><p>In summary, I have identified a subset of miRNAs, the expression of which can be regulated by SF2/ASF in a splicing independent manner. This is the first SR protein identified in regulating miRNA biogenesis. One of the upregulated miRNAs, miRNA-7 can form a negative feedback with SF2/ASF by negatively regulating the expression of SF2/ASF on translational level. By using PAR-CLIP method, I have identified the genome-wide binding partners of SRp20 in vivo. When SRp20 binds to the exonic regions, it potentially affects the alternative splicing patterns of nearby introns. Interestingly, the 3' end choices for a subset of genes may be regulated by SRp20 through directly binding, which may be a new mechanism for the regulation of 3' end processing.</p> / Dissertation

FlexRay is a new automotive network communication protocol for control and interconnection among ECUs (electronic control units) in the cluster. In the FlexRay protocol, a communication cycle consists of static segment and dynamic segment. The static segment is a TDMA scheme designed for transmitting time-triggered messages. Due to its determinism and reliability, it is particularly applicable to X-by-wire applications. Each static slot is allocated to a specified task and the task can transmit message during the exclusive slot. However, if the task has no message to transmit during its assigned slot, the slot cannot be used by other tasks. The overall utilization is low if the bandwidth requirement of each task is not high. To improve the system utilization, we apply the real-time scheduling techniques to devising a deterministic, static cyclic scheduling. The objective is to reduce the demand on the number of static slots needed for scheduling time-triggered tasks. Specifically, we treat the set of static slots that are in the same position in every communication cycle as an individual real-time channel. We model each task as a real-time task, specified by (Ci,Ti). It requires that for every Ti communication cycles, the system must allocate at least Ci time slots to satisfy the real-time constraint of the task. We decompose each such task into a set of subtasks, allocate them to the real-time channels and then apply the rate-monotonic scheduling algorithm to schedule the subtasks within each channel. Finally, we perform computer simulation to evaluate the effectiveness of our proposal. From the simulation results, we conclude that our proposal is able to effectively reduce the demand for the static slots under a wide range of real-time requirements.

Accurate forecasts of future regional temperature and rainfall patterns in many regions largely depend on characterizing anthropogenic trends in tropical Pacific climate. However, strong interannual to decadal-scale tropical Pacific climate variability, combined with sparse spatial and temporal coverage of instrumental climate datasets in this region, have obscured potential anthropogenic climate signals in the tropical Pacific. In this dissertation, I present sea-surface temperature (SST) and salinity proxy records that span over the 20th century using living corals from several islands in the central tropical Pacific. I reconstruct the SST proxy records via coral Sr/Ca, that are combined with coral oxygen isotopic (d18O) records to quantify changes in seawater d18O (hereafter d18Osw) as a proxy for salinity. Chapter 2 investigates the spatial and temporal character of SST and d18Osw-based salinity trends in the central tropical Pacific from 1972-1998, as revealed by corals from Palmyra (6ºN, 162ºW), Fanning (4ºN, 159ºW) and Christmas (2ºN, 157ºW) Islands. The late 20th century SST proxy records exhibit warming trends that are larger towards the equator, in line with a weakening of equatorial Pacific upwelling over this period. Freshening trends revealed by the salinity proxy records are larger at those sites most affected by the Inter-Tropical Convergence Zone (ITCZ), suggesting a strengthening and/or an equatorward shift of the ITCZ. Taken together, the late 20th century SST and salinity proxy records document warming and freshening trends that are consistent with a trend towards a weakened tropical Pacific zonal SST gradient under continued anthropogenic forcing. Chapter 3 characterizes the signatures of natural and anthropogenic variability in central tropical Pacific SST and d18Osw-based salinity over the course of 20th century using century-long coral proxy records from Palmyra. On interannual timescales, the SST proxy record from Palmyra tracks El Niño-Southern Oscillation (ENSO) variability. The salinity proxy record tracks eastern Pacific-centered ENSO events but is poorly correlated to central Pacific-centered ENSO events - the result of profound differences in precipitation and ocean advection that occur during the two types of ENSO. On decadal timescales, the coral SST proxy record is significantly correlated to the North Pacific Gyre Oscillation (NPGO), suggesting that strong dynamical links exist between the central tropical Pacific and the North Pacific. The salinity proxy record is significantly correlated to the Pacific Decadal Oscillation (PDO), but poorly correlated to the NPGO, suggesting that, as was the case with ENSO, these two modes of Pacific decadal climate variability have unique impacts on equatorial precipitation and ocean advection. However, the most striking feature of the salinity proxy record is a prominent late 20th century freshening trend that is likely related to anthropogenic climate change. Taken together, the coral data provide key constraints on tropical Pacific climate trends, and when used in combination with model simulations of 21st century climate, can be used to improve projections of regional climate in areas affected by tropical Pacific climate variability.

Il existe actuellement deux structures nanotubulaires largement étudiées : les nanotubes de carbone (CNTs) possédant un gap de l'ordre de 1 eV et ceux de nitrure de bore (BN-NTs) dont le gap est compris entre 5 et 6 eV. Parvenir à doper ces nanotubes par substitution d'éléments chimiques pouvant introduire des états donneurs ou accepteurs est supposé être une approche prometteuse pour moduler les propriétés électroniques et optiques de ces nanostructures. Les objectifs de cette thèse ont été doubles. D'une part, il s'est agi de développer des procédés de synthèse modulables pour pouvoir produire différents types de tubes hétéroatomiques. La technique CVD assistée par aérosol et la technique de vaporisation laser ont été utilisées à cette fin. D'autre part, une analyse fine à l'échelle macroscopique et sub-nanométrique des échantillons a été réalisée principalement par microscopie électronique en transmission (HRTEM) et spectroscopie de pertes d'énergies des électrons résolue spatialement (SR-EELS).<br />Il a ainsi été mis en évidence dans ce manuscrit la possibilité de réaliser des nanotubes de type CNx et CBxNy mono- et multi-feuillets. Par ailleurs, une forte tendance à la ségrégation en domaines de type C et BN a été observée avec un localisation préférentielle et une taille des domaine spécifique, fonction de la technique de synthèse utilisée.

Title from PDF of title page. / Document formatted into pages; contains 93 pages / Thesis (M.S.)--University of South Florida, 2003. / Includes bibliographical references. / Text (Electronic thesis) in PDF format. / ABSTRACT: Coral-based climate studies commonly use elemental ratios and stable isotopes of coral skeletons to address seawater temperature and hydrologic balance issues in the tropical surface oceans. Replication, or cross-checking, a standard technique used to assess the fidelity of proxy records in paleoclimatology has not been widely applied in coral-based climate studies, primarily because of the time and cost associated with generating multiple records from a single reef site. Modern and paleoclimate reconstructions based on a single proxy-coral record from a site may contain errors if individual corals from the same reef record different geochemical signals. In this study we perform a replication test using elemental ratios and stable isotopes in three Porites corals from New Caledonia. / ABSTRACT: The reef complex offshore Amédée Island, New Caledonia is an ideal site to perform a coral replication test because instrumental sea surface temperatures (SST) and sea surface salinity measurements (SSS) have been made there for over 25 years. In this study, we compare sub-monthly resolved, geochemical variations (Sr/Ca, d18O and d13C) in three Porites lutea coral heads, located 500 m apart, with the instrumental SST and SSS records over the interval 1992-1967. The monthly coral Sr/Ca and d18O time series are well correlated to each other (r=0.86, p[.0001) and to the monthly instrumental SST record (r= -0.86, p[.0001, coral Sr/Ca to SST; r= -0.77, p[.0001, coral d18O to SST). The three, sub-monthly resolved, 30-year coral Sr/Ca-SST time series have mean SST values that agree within 0.2oC with the instrumental mean SST value. A similar comparison for the coral d18O-SST records indicate a maximum difference between predicted and observed mean SST of 0.5°C. / ABSTRACT: Analysis of the monthly climatological means also indicates that Sr/Ca-SST records closely match the instrumental SST record ±0.4°C; a similar comparison using the d18O-SST record yields an average offset of ±0.6°C between observed and predicted monthly SST. Stacking the three records to form composite Sr/Ca-SST and d18O-SST records does not appreciably improve the goodness of fit between the proxy and instrumental SST records; hence a coral-based proxy climate record from a single coral accurately reflects the observed record of climate variability at this locality. These results support the concept that high fidelity climate records can be generated using a single coral core. / System requirements: World Wide Web browser and PDF reader. / Mode of access: World Wide Web.

Aluminium alloys with Si as the major alloying element form a class of material providing the most significant part of all casting manufactured materials. These alloys have a wide range of applications in the automotive and aerospace industries due to an excellent combination of castability and mechanical properties, as well as good corrosion resistance and wear resistivity. Additions of minor alloying elements such as Cu and Mg improve the mechanical properties and make the alloy responsive to heat treatment. The aim of this work is studying the role of size and morphology of microstructural constituents (e.g SDAS, Si-particles and intermetalics) on mechanical properties of Al-Si based casting alloy at room temperatures up to 500 ºC. The cooling rate controls the secondary dendrite arm spacing (SDAS), size and distribution of secondary phases. As SDAS becomes smaller, porosity and second phase constituents are dispersed more finely and evenly. This refinement of the microstructure leads to substantial improvement in tensile properties (e.g. Rm and εF). Addition of about 280 ppm Sr to EN AC- 46000 alloy yields fully modified Si-particles (from coarse plates to fine fibres) regardless of the cooling conditions. Depression in eutectic growth temperature as a result of Sr addition was found to be strongly correlated to the level of modification irrespective of coarseness of microstructure. Modification treatment can improve elongation to failure to a great extent as long as the intermetallic compounds are refined in size. Above 300 ºC, tensile strength, Rp0.2 and Rm, of EN AC-46000 alloys are dramatically degraded while the ductility was increased. The fine microstructure (SDAS 10 μm) has superior Rm and ductility compared to the coarse microstructure (SDAS 25 μm) at all test temperature (from room to 500 ºC). Concentration of solutes (e.g. Cu and Mg) in the dendrites increases at 300 ºC and above where Rp0.2 monotonically decreased. The brittleness of the alloy below 300 ºC was related to accumulation of a high volume fraction damaged particles such as Cu- Fe-bearing phases and Si-particles. The initiation rate of damage in the coarse particles was significantly higher, which enhances the probability of failure and decreasing both Rm and εF compared to the fine microstructure. A physically-based model was adapted, improved and validated in order to predict the flow stress behaviour of EN AC- 46000 cast alloys at room temperature up to 400 ºC for various microstructures. The temperature dependant variables of the model were quite well correlated to the underlying physics of the material

The Ertsberg-Grasberg Mining District of Papua, Indonesia (Western New Guinea) hosts the Ertsberg Cu-Au Skarn, the giant Grasberg Porphyry Cu-Au deposit, and several other orebodies. Two 1700-meter-long cores beneath the Kucing Liar ore skarn (KL98-10-22) and the Grasberg Igneous Complex (KL98-10-21) contain high concentrations of vein and disseminated molybdenite. KL98-10-22, the focus of this study, intersects two previously unencountered intrusions, the “Tertiary intrusion Kucing Liar” (Tikl) and “Tertiary Pliocene intrusion” (Tpi). An intense dilatational quartz vein stockwork cuts Tikl and Ekmai Sandstone (Kkes) units, predating Tpi intrusion. Prior to these ultradeep cores, which extend almost 3 km below pre-mining surface, molybdenite was rarely observed in the district. Geochemistry and isotopic data indicate that Tikl and Tpi intrusions originated from the same large magmatic system that emplaced other ore-forming Ertsberg-Grasberg district intrusions. Magma in a lower crustal chamber was recharged at least twice, according to Sr-Nd data. Laser-ablation inductively-coupled plasma mass spectrometry of magmatic zircons yields 238U-206Pb ages between 3.40 ± 0.12 Ma (Dalam Andesite) and 2.77 ± 0.15 Ma (Ertsberg intrusion), revealing a shorter period of igneous activity than previously measured by K-Ar and Ar-Ar dating. Analyses include composite ages of 3.28 ± 0.08 Ma for Tikl and 3.18 ± 0.11 Ma for Tpi. Inherited zircon cores indicate Precambrian (mostly Proterozoic) basement. Molybdenite veining beneath the Kucing Liar Skarn and Grasberg Igneous Complex postdates stockwork veining and occurred before the 2.99 ± 0.11 Ma Kali dikes. Only one molybdenite vein was observed cutting Tpi. Molybdenites yielded ~3 Ma Re-Os ages and anomalous >4 Ma and <0.5 Ma ages; anomalous ages were not reproducible in follow-up analyses (this study). Smearing deformation of molybdenite (through fault activity) causes crystal strain, likely leading to annealing recrystallization. Recrystallization possibly redistributes daughter-product Os, resulting in anomalous ages from annealed material. Fluids with high Mo/Cu ratios (which were likely supercritical) precipitated late-stage molybdenite deep in the system. These fluids developed through magma chamber crystallization, which concentrated molybdenum in the melt as an incompatible element, and stripping of Cu from the magma chamber during hydrothermal activity. / text