Minimum requirements for lap splices in reinforced concrete members, stated in building codes of TS-500 and ACI-318, have a certain factor of safety. These standards have been prepared according to research results conducted previously and they are being updated according to results of recent studies. However the reliability of lap splices for minimum requirements needs to be investigated. For this purpose, 6 beam specimens were prepared according to minimum provisions of these standards. The test results were investigated by analytical procedures and also a parametric study was done to compare two standards. For smaller diameter bars both standards give safe results. Results showed that the minimum clear cover given in TS500 is insufficient for lap spliced bars greater than or equal to 26 mm diameter.

Behavior of lap splices formed by CFRP rolls has been studied. CFRP rolls have been prepared by using CFRP sheets of a certain width. Strengthening methods that use CFRP rolls as reinforcement may require an epoxy anchored lap splice due to the conditions at the strengthening regions. It may not always be possible to strengthen the region by using only one roll fan anchored at both ends, but using two rolls from opposite faces of the member and lap splicing them at the middle so that they act as a single roll. Lap splice behavior can be studied best by using flexural beam bond specimens if the reinforcing material is steel. Therefore, it has initially been suggested that flexural beam specimens reinforced for flexure with CFRP rolls as tension reinforcement can be used in studying the lap splice behavior. However, due to the difficulties encountered in the beam tests, another type of test specimen was introduced, which was a direct pull-out specimen. In this type of test specimen, lap spliced CFRP rolls have been tested under direct tension, in which the tension has been applied by making use of concrete end blocks that transfer the tension to the rolls. Eleven tests have been made in total. Full material capacity of the rolls could not be achieved due to premature failures. However, important conclusions and recommendations have been made for future studies.

Spliced girder technology continues to attract attention due to its versatility over traditional prestressed concrete highway bridge construction. Relatively limited data is available in the literature, however, for large-scale tests of post-tensioned I-girders, and few studies have examined the behavior of the cast-in-place (CIP) splice regions of post-tensioned spliced girder bridges. In addition to limited knowledge on CIP splice region behavior, a wide variety of splice region details (e.g., splice region length, mild reinforcement details, cross-sectional geometry, etc.) continue to be used in the field. In response to these issues, the research program described in this dissertation was developed to (i) study the strength and serviceability behavior of the CIP splice regions of spliced I-girders, (ii) identify design and detailing practices that have been successfully implemented in CIP splice regions, and (iii) develop design recommendations based on the structural performance of spliced I-girder test specimens. To accomplish these tasks, an industry survey was first conducted to identify the best practices that have been implemented for the splice regions of existing bridges. Splice region details were then selected to be included in large-scale post-tensioned spliced I-girder test specimens. Two tests were conducted to study splice region behavior and evaluate the performance of the chosen details. The failure mechanisms of both test girders were characterized by a shear-compression failure of the web concrete with primary crushing occurring in the vicinity of the top post-tensioning duct. Most significantly, the girders acted essentially as monolithic members in shear at failure. Web crushing extended across much of the test span and was not localized within the splice regions. To supplement the spliced girder tests, a shear-friction experimental program was also conducted to gain a better understanding of the interface shear behavior between precast and CIP concrete surfaces at splice regions. The findings of the shear-friction study are summarized within this dissertation. Based on the results of the splice region research program, design recommendations were developed, including recommended CIP splice region details.

The hydrophobic plasma membrane constitutes an indispensable barrier for cells in living animals. Albeit being pivotal for the maintenance of cells, the inability to cross the plasma membrane is still one of the major obstacles to overcome in order to progress current drug development. A group of substances, with restricted access to the interior of cells, which has shown great promise for future clinical use is oligonucleotides that are exploited to interfere with gene expression. Short interfering RNAs that are utilized to confer gene silencing and splice correcting oligonucleotides, applied for the manipulation of splicing patterns, are two classes of oligonucleotides that have been explored in this thesis. Cell-penetrating peptides (CPPs) are a class of peptides that has gained increasing focus in last years. This ensues as a result of their remarkable ability to convey various, otherwise impermeable, macromolecules across the plasma membrane of cells in a relatively non-toxic fashion. This thesis aims at further characterizing well-established, and newly designed, CPPs in terms of toxicity, delivery efficacy, and internalization mechanism. Our results demonstrate that different CPPs display different toxic profiles and that cargo conjugation alters the toxicity and uptake levels. Furthermore, we confirm the involvement of endocytosis in translocation of CPPs, and in particular the importance of macropinocytosis. All tested peptides facilitate the delivery of splice correcting oligonucleotides with varying efficacy, the newly designed CPP, M918, being the most potent. Finally we conclude that by promoting endosomolysis, by exploring new CPPs with improved endosomolytic properties, the biological response increases significantly. In conclusion, we believe that these results will facilitate the development of new CPPs with improved delivery properties that could be used for transportation of oligonucleotides in clinical settings.

Bridge expansion joints are directly subjected to traffic load, and thus prone to premature fatigue failure. Replacement of components such as modular bridge expansion joints is typically done in a staggered schedule to minimize traffic blockage. Field splices are used to connect the successively installed segments. These splices typically include a combination of field welding or bolting, and experience has shown that they often fail due to fatigue cracking. This thesis reports the investigation of hybrid FRP/steel splice details that avoid the use of field welding. Two configurations have been examined: A GFRP pultruded square tube section, adhesively bonded to the soffit of the spliced beam, consists the moment resisting component in one configuration, whereas the other takes advantage of two series of FRP plates for this purpose. Bolted steel plates splice the beam through web in both cases. The behaviour of these details has been studied extensively under vertical static loads. The effect of several parameters including bond length, FRP end shape, bond surface treatment, adhesive, etc. for each detail has been investigated. A three-dimensional, non-linear finite element model has been developed for each detail and validated using the experimental results. The bond strength of two adhesives was investigated experimentally using double shear lap splice tests. A new method is proposed to analyze the strength of the splice details. This method is based on the results obtained from shear lap splice tests and the verified finite element model developed for the splice detail. The finite element model could thus be used for further parametric studies. More experiments, however, are statistically required before using this model with confidence. The fatigue behaviour of one of the promising splice details has been investigated both experimentally and numerically. A special fatigue test set-up has been designed and used successfully for this purpose. Two fatigue tests to 1,000,000 cycles were run. One failed at 719, 347 cycles and the other survived 1,000,000 cycles. The predicted fatigue life as per the developed model was 871,840 cycles. More experiments are required to understand the fatigue behaviour of the splice detail under various stress ranges. / Thesis (Ph.D, Civil Engineering) -- Queen's University, 2011-11-30 16:53:07.385

Group I introns belong to a class of large RNAs that catalyze their own excision from precursor RNA through a two-step process called self-splicing reaction. These self-splicing introns have often been converted into ribozymes with the ability site specifically cleave RNA molecules. One such ribozyme, derived from a self-splicing Pneumocystis carinii group I intron, has subsequently been shown to sequence specifically excise a segment from an exogenous RNA transcript through trans excision-splicing reaction.The trans excision-splicing reaction requires that the substrate be cleaved at two positions called the 5' and 3' splice sites. The sequence requirements at these splice sites were studied. All sixteen possible base pair combinations at the 5' splice site and the four possible nucleotides at the 3' splice site were tested for reactivity. It was found that all base pair combinations at the 5' splice site allow the first reaction step and seven out of sixteen combinations allow the second step to occur. Moreover, it was also found that non-Watson-Crick base pairs are important for 5' splice site recognition and suppress cryptic splicing. In contrast to the 5' splice site, 3' splice site absolutely requires a guanosine.The pathway of the trans excision-splicing reaction is poorly understood. Therefore, as an initial approach, a kinetic framework for the first step (5' cleavage) was established. The framework revealed that substrate binds at a rate expected for RNA-RNA helix formation. The substrate dissociates with a rate constant (0.9 min-1), similar to that for substrate cleavage (3.9 min-1). Following cleavage, the product dissociation is slower than the cleavage, making this step rate limiting for multiple-turnover reactions. Furthermore, evidence suggests that P10 helix forms after the 5' cleavage step and a conformational change exists between the two reaction steps of trans excision-splicing reaction. Combining the data presented herein and the prior knowledge of RNA catalysis, provide a much more detailed view of the second step of the trans excision-splicing reaction.These studies further characterize trans excision-splicing reaction in vitro and provide an insight into its reaction pathway. In addition, the results describe the limits ofthe trans excision-splicing reaction and suggest how key steps can be targeted for improvement using rational ribozyme design approach.

BAI2 gehört zu den Adhesion-G-Protein-gekoppelten Rezeptoren (aGPCR). Diese bisher wenig untersuchte Klasse von ca. 30 GPCR ist charakterisiert durch eine komplexe genomische Struktur, sehr große extrazelluläre Domänen und eine Vielzahl von Splicevarianten. Bisher ist bei den meisten aGPCR, wie auch bei BAI2, wenig über ihre Signaltransduktion und Funktion bekannt. Zum Verständnis der physiologischen Relevanz und zur Suche nach dem endogenen Agonist sind Kenntnisse über Proteinstruktur, Splicevarianten und Signaltransduktion essentiell. Ziel dieser Arbeit war es, mittels verschiedener in vitro-Methoden die Proteinstruktur des BAI2 in den transmembranären und intrazellulären Domänen näher zu untersuchen, sowie die natürlichen Splicevarianten in diesem Bereich, deren evolutionäre Konservierung, Gewebespezifität und Quantität zu erfassen. Für beide gefundenen Splicevarianten, eine im dritten intrazellulären Loop (ICL3) und eine im C-Terminus, konnte eine evolutionäre Konservierung auf Aminosäure- und genomischer Organisationsebene, sowie ihre Entstehung durch Exonskipping nachgewiesen werden. Nachfolgend wurden die Splicevarianten auf mögliche Interaktionen mit intrazellulären Komponenten untersucht. In dieser Arbeit konnte gezeigt werden, dass beide ICL3-Splicevarianten natürlicherweise in einem definierten Verhältnis auftreten. Außerdem konnte gezeigt werden, dass die lange ICL3-Variante des BAI2 nicht zu einer Änderung der Membrantopologie des Rezeptors, einer Homodimerisierung über die zusätzliche Aminosäuresequenz oder zu einer Interaktion mit dem C-Terminus führt. Die Splicevariante im humanen C-Terminus des BAI2 konnte als eine variable, durch Exonskipping entstandene Calcium-unabhängige Calmodulin-Bindungsstelle identifiziert werden. Diese Arbeit belegt die Existenz mehrerer BAI2-Isoformen in vivo. Die Struktur dieser Isoformen lässt unterschiedliche Funktionalitäten vermuten. Auch wenn erste Untersuchungen zwischen den beiden ICL3-Varianten keinen Unterschied ergaben, sind diese Erkenntnisse für die weitere Analyse der Signaltransduktion und Ligandensuche bedeutend. Es ist z.B. denkbar, dass sich die beiden ICL3-Varianten in der G-Protein-Kopplung oder bei der Rekrutierung von intrazellulären Interaktionspartnern unterscheiden oder dass die Splicevariante im C-Terminus zu einer Scaffold- Funktion des Calmodulins führt und/oder die Signaltransduktion durch eine permanente Bindung des Calmodulins an einer Isoform moduliert wird.

No description available.

Thesis (Ph.D.)--Indiana University, 2009. / Title from screen (viewed on August 28, 2009). Department of Biochemistry and Molecular Biology, Indiana University-Purdue University Indianapolis (IUPUI). Advisor(s): William Bosron. Includes vita. Includes bibliographical references (leaves 102-111).

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 106-119).