The I-35W Bridge over the Mississippi River in Minneapolis, MN had a catastrophic failure in the main span of the deck truss in 2007. This collapse has brought significant attention on the gusset plate connections in steel truss bridges throughout the U.S. Steel truss bridge gusset plate design has not received much focus in the past 40 years, and there is a lack of consensus within the design profession on the procedures to evaluate, design, and rate these critical elements. In the short term, based on the best available information on the gusset plate design, the Federal Highway Administration (FHWA) has issued preliminary guidance. Although some experimental research has been conducted on the ultimate strength of gusset plates, much of this work has been directed toward the performance of tension members and their connections. There has been limited experimental work on the compression capacity and stability of gusset plates, but most of this work is relevant primarily to bracing connections common in building structures. This research focuses on comprehensive experimental and analytical studies on steel truss bridge gusset plate behavior. The studies include comparisons of advanced analytical models with the responses from large-scale experimental tests using discrete and innovative full-field measurements. The calibrated finite element analysis models are then utilized to study a variety of gusset plate configurations. Improved mechanistic idealizations that better capture the observed behavior in the experiments and analytical studies are proposed as the result of this work. The design checks recommended in this thesis present a comprehensive methodology for determining the ultimate gusset plate resistance. This research provides a large database of original results that will be useful for future similar studies. In addition, this research provides modeling procedures that permit the study of steel truss bridge connections and their adjacent framing members using truss bridge sub-assemblies. Based on the comprehensive analytical studies, simple and accurate design calculation procedures to assess the nominal ultimate strength of steel truss gusset plate connections are recommended for steel truss bridge gusset plate connections.

VASA ist ein DEAD-Box-Protein, das in der Keimbahn vorkommt und dem eine essentielle Rolle in der Entstehung und Erhaltung von Keimzellen zugeschrieben wird (Yajima und Wessel 2011a). VASA wurde in jedem bisher untersuchten Organismus in der Keimbahn gefunden (Raz 2000) und ist, da es in Säugetieren als keimzellspezifisch gilt (Castrillon et al. 2000), einer der derzeit meist genutzten Keimzellmarker. Allerdings zeigen neuere Daten aus unkonventionellen Modellorganismen sowie aus Drosophila, dass VASA in diesen Organismen, zusätzlich zu seinen Funktionen in der Keimbahn, noch weitere Funktionen außerhalb der Keimbahn übernimmt. Bisher existieren allerdings keine vergleichbaren Daten, die die Keimzellspezifität von VASA in Säugetieren widerlegen, weshalb VASA hier weiter-hin als spezifischer Keimzellmarker gilt (Alié et al. 2011). Vorarbeiten von Selma Drallé, die in der Arbeitsgruppe von Prof. Dr. rer. nat. R. Behr durchgeführt wurden, ergaben jedoch Hinweise auf eine VASA-Expression in somatischen Zellen des Weißbüschelaffen. In Weiterführung dieser Untersuchungen wurde in der vorliegenden Arbeit eine systematische Expressionsanalyse von VASA in unterschiedlichen Organen des adulten und neugeborenen Weißbüschelaffen durchgeführt. Die Arbeitshypothese hierbei war, dass VASA im Weischbüschelaffen nicht keimzellspezifisch exprimiert wird. Die Untersuchungen erfolgten mittels immunhistochemischer Färbungen, Western Blotting sowie PCR, wobei aufgrund präliminarer Hinweise jeweils Niere, Magen, Leber, Haut und Pankreas des neugeborenen und des adulten Weißbüschelaffen detailliert auf eine VASA-Expression untersucht wurden. Ovar und Hoden adulter Weißbüschelaffen dienten als Positivkontrollen. Die Ergebnisse dieser Arbeit deuten stark auf eine Expression von VASA in der Haut des neugeborenen Weißbüschelaffen hin. Dieser Befund legt nahe, das VASA im neugeborenen Weißbüschel-affen nicht keimzellspezifisch ist und über seine Funktionen in der Keimbahn hinaus noch weitere extragonadale Funktionen innehaben könnte. In den weiteren untersuchten Organen wurde keine VASA-Expression festgestellt; einzelne Befunde, die auf eine VASA-Expression hinwiesen, erwiesen sich als falsch-positiv. In dieser Arbeit wurde zudem eine für C. jacchus noch nicht in der Literatur beschriebene Splice-Variante von VASA im Ovar und im Hoden vom adulten Weißbüschelaffen gefunden, der das Exon 7 fehlt und die zudem modifizierte N- und/oder C-Termini aufweisen könnte. Die vorliegenden Ergebnisse weisen darauf hin, dass VASA im Weißbüschelaffen auch außerhalb der Keimbahn exprimiert wird und somit in dieser Spezies kein spezifischer Keim-zellmarker ist. Um diese Frage jedoch abschließend zu klären, sollten sich weitere Untersuchungen anschließen, um die in dieser Arbeit gezeigten Befunde zu untermauern.

The aim of this study is to analyse how pupils in grade three use punctuation and howthey divide texts into sentences. The material in the study consists of 41 texts by pupils,divided into two types: narrative text and analytical text, written both by pupils withSwedish as their first language and by pupils with Swedish as their second language. Inthe study the pupils’ use of connectives and comma splices has been studied, and therelationship between graphical and syntactical sentences in the texts. The studycompares the results for narrative and analytical texts and the differences between L1and L2 pupils. The study is a follow-up of Per Ledin’s (1998) study of pupils’ use ofpunctuation in the low and intermediate levels of compulsory, and therefore the resultsof this study are compared with those obtained by Ledin. This study shows that in thisrespect L1 and L2 pupils produce texts that do not differ to any great extent. The mostprominent differences between the types of text are that the narrative texts have aclearer flow and a seemingly more logical link between sentences and clauses than theanalytical texts. The results of this study largely agree with Ledin’s results.

No description available.

A plethora of naturally-produced steroid hormones, or artificial homologues of them, are being introduced into the aquatic and terrestrial environments each year. Two examples of these are the natural oestrogen 17-oestradiol (E2) and the oestrogen receptor antagonist, Bisphenol A (BPA), both of which target the ribonucleoprotein telomerase through upregulation of its telomerase reverse transcriptase component, TERT. The main objectives of this study were firstly to isolate and characterize the actual mRNA sequence for the telomerase catalytic subuninit, Tert, in rainbow trout (Oncorhynchus mykiss) (Walbaum, 1792) and European purple sea urchin (Paracentrotus lividus) (Lamarck, 1816), with the aim of developing qPCR assays for the amplification and quantification of Tert. Further objectives were to use these assays in controlled exposure studies to establish whether and to what extent the aforementioned chemicals regulate Tert transcription and by doing so further understand the mechanism of Telomerase gene expression and the extent to which environmental oestrogen can interfere. The initial step of sequence characterization and assay devlopment was successful in the case of rainbow trout where two possible splice variants of Tert mRNA are identified, omTertShort and omTertLong. Two qPCR assays were developed for the relative quantification of both of these splice variants in rainbow trout samples, the latter of these successfully amplifying its target in test samples. In order to demonstrate in vitro and in vivo modulation of telomerase activity and mRNA expression, early life-stages of rainbow trout and purple sea urchin, as well as rainbow trout hepatocytes, were exposed to a range of concentrations of E2 and BPA. Purple sea urchin embryos were exposed to 200, 20 and 2 ng E2/ml for 28 hours until they had reached the stage of pluteus larvaes. Rainbow trout embryos were exposed to 500, 20 and 0.1 ng E2/ml and 600 and 150 ng BPA/ml for 167 days from immediately after fertilization. Rainbow trout hepatocytes were exposed to 20 and 2 ng E2/ml for 48 hours. The results from this study show that telomerase activity as well as TERT mRNA expression can be significantly modulated by exposure to oestrogens and other oestrogenic chemicals. E2 concentrations as low as 20 ng/ml lead to an increase in telomerase activity early-life stages of purple sea urchin and upregulation in the transcription of Tert mRNA in unhatched rainbow trout embryos. BPA induced similar response (600 ng/ml) in hatched rainbow trout alevins larvae. Very high exposures to E2 (500 ng/ml) do however lead to downregulation of Tert mRNA in hatched alevins larvae. Differential regulatory response can be observed between different tissue types of 167 day old fry, with an upregulatory response observed at 0.1 ng E2/ml in liver and muscle tissues, but not in brain. Similarly, brain tissues were observed expressing significantly less mRNA than liver and muscle samples when exposed to BPA (150 ng/ml). It is evident that the previously observed link between environmental oestrogens and telomerase is also present in the two test species examined; purple sea urchin and rainbow trout.

L'incessant augment del nombre de seqüències genòmiques, juntament amb l'increment del nombre de tècniques experimentals de les que es disposa, permetrà obtenir el catàleg complet de les funcions cel.lulars de diferents organismes, incloent-hi la nostra espècie. Aquest catàleg definirà els fonaments sobre els que es podrà entendre millor com els organismes funcionen a nivell molecular. Al mateix temps es tindran més pistes sobre els canvis que estan associats amb les malalties. Per tant, la seqüència en brut, tal i com s'obté dels projectes de seqüenciació de genomes, no té cap valor sense les anàlisis i la subsegüent anotació de les característiques que defineixen aquestes funcions. Aquesta tesi presenta la nostra contribució en tres aspectes relacionats de l'anotació dels gens en genomes eucariotes. Primer, la comparació a nivell de seqüència entre els genomes humà i de ratolí es va dur a terme mitjançant un protocol semi-automàtic. El programa de predicció de gens SGP2 es va desenvolupar a partir d'elements d'aquest protocol. El concepte al darrera de l'SGP2 és que les regions de similaritat obtingudes amb el programa TBLASTX, es fan servir per augmentar la puntuació dels exons predits pel programa geneid, amb el que s obtenen conjunts d'anotacions més acurats d'estructures gèniques. SGP2 té una especificitat que és prou gran com per que es puguin validar experimentalment via RT-PCR. La validació de llocs d'splicing emprant la tècnica de la RT-PCR és un bon exemple de com la combinació d'aproximacions computacionals i experimentals produeix millors resultats que per separat. S'ha dut a terme l'anàlisi descriptiva a nivell de seqüència dels llocs d'splicing obtinguts sobre un conjunt fiable de gens ortòlegs per humà, ratolí, rata i pollastre. S'han explorat les diferències a nivell de nucleòtid entre llocs U2 i U12, pel conjunt d'introns ortòlegs que se'n deriva d'aquests gens. S'ha trobat que els senyals d'splicing ortòlegs entre humà i rossegadors, així com entre rossegadors, estan més conservats que els llocs no relacionats. Aquesta conservació addicional pot ser explicada però a nivell de conservació basal dels introns. D'altra banda, s'ha detectat més conservació de l'esperada entre llocs d'splicing ortòlegs entre mamífers i pollastre. Els resultats obtinguts també indiquen que les classes intròniques U2 i U12 han evolucionat independentment des de l'ancestre comú dels mamífers i les aus. Tampoc s'ha trobat cap cas convincent d'interconversió entre aquestes dues classes en el conjunt d'introns ortòlegs generat, ni cap cas de substitució entre els subtipus AT-AC i GT-AG d'introns U12. Al contrari, el pas de GT-AG a GC-AG, i viceversa, en introns U2 no sembla ser inusual. Finalment, s'han implementat una sèrie d'eines de visualització per integrar anotacions obtingudes pels programes de predicció de gens i per les anàlisis comparatives sobre genomes. Una d'aquestes eines, el gff2ps, s'ha emprat en la cartografia dels genomes humà, de la mosca del vinagre i del mosquit de la malària, entre d'altres. El programa gff2aplot i els filtres associats, han facilitat la tasca d'integrar anotacions de seqüència amb els resultats d'eines per la cerca d'homologia, com ara el BLAST. S'ha adaptat també el concepte de pictograma a l'anàlisi comparativa de llocs d splicing ortòlegs, amb el desenvolupament del programa compi. / El aumento incesante del número de secuencias genómicas, junto con el incremento del número de técnicas experimentales de las que se dispone, permitirá la obtención del catálogo completo de las funciones celulares de los diferentes organismos, incluida nuestra especie. Este catálogo definirá las bases sobre las que se pueda entender mejor el funcionamiento de los organismos a nivel molecular. Al mismo tiempo, se obtendrán más pistas sobre los cambios asociados a enfermedades. Por tanto, la secuencia en bruto, tal y como se obtiene en los proyectos de secuenciación masiva, no tiene ningún valor sin los análisis y la posterior anotación de las características que definen estas funciones. Esta tesis presenta nuestra contribución a tres aspectos relacionados de la anotación de los genes en genomas eucariotas. Primero, la comparación a nivel de secuencia entre el genoma humano y el de ratón se llevó a cabo mediante un protocolo semi-automático. El programa de predicción de genes SGP2 se desarrolló a partir de elementos de dicho protocolo. El concepto sobre el que se fundamenta el SGP2 es que las regiones de similaridad obtenidas con el programa TBLASTX, se utilizan para aumentar la puntuación de los exones predichos por el programa geneid, con lo que se obtienen conjuntos más precisos de anotaciones de estructuras génicas. SGP2 tiene una especificidad suficiente como para validar esas anotaciones experimentalmente vía RT-PCR. La validación de los sitios de splicing mediante el uso de la técnica de la RT-PCR es un buen ejemplo de cómo la combinación de aproximaciones computacionales y experimentales produce mejores resultados que por separado. Se ha llevado a cabo el análisis descriptivo a nivel de secuencia de los sitios de splicing obtenidos sobre un conjunto fiable de genes ortólogos para humano, ratón, rata y pollo. Se han explorado las diferencias a nivel de nucleótido entre sitios U2 y U12 para el conjunto de intrones ortólogos derivado de esos genes. Se ha visto que las señales de splicing ortólogas entre humanos y roedores, así como entre roedores, están más conservadas que las no ortólogas. Esta conservación puede ser explicada en parte a nivel de conservación basal de los intrones. Por otro lado, se ha detectado mayor conservación de la esperada entre sitios de splicing ortólogos entre mamíferos y pollo. Los resultados obtenidos indican también que las clases intrónicas U2 y U12 han evolucionado independientemente desde el ancestro común de mamíferos y aves. Tampoco se ha hallado ningún caso convincente de interconversión entre estas dos clases en el conjunto de intrones ortólogos generado, ni ningún caso de substitución entre los subtipos AT-AC y GT-AG en intrones U12. Por el contrario, el paso de GT-AG a GC-AG, y viceversa, en intrones U2 no parece ser inusual. Finalmente, se han implementado una serie de herramientas de visualización para integrar anotaciones obtenidas por los programas de predicción de genes y por los análisis comparativos sobre genomas. Una de estas herramientas, gff2ps, se ha utilizado para cartografiar los genomas humano, de la mosca del vinagre y del mosquito de la malaria. El programa gff2aplot y los filtros asociados, han facilitado la tarea de integrar anotaciones a nivel de secuencia con los resultados obtenidos por herramientas de búsqueda de homología, como BLAST. Se ha adaptado también el concepto de pictograma al análisis comparativo de los sitios de splicing ortólogos, con el desarrollo del programa compi. / The constantly increasing amount of available genome sequences, along with an increasing number of experimental techniques, will help to produce the complete catalog of cellular functions for different organisms, including humans. Such a catalog will define the base from which we will better understand how organisms work at the molecular level. At the same time it will shed light on which changes are associated with disease. Therefore, the raw sequence from genome sequencing projects is worthless without the complete analysis and further annotation of the genomic features that define those functions. This dissertation presents our contribution to three related aspects of gene annotation on eukaryotic genomes. First, a comparison at sequence level of human and mouse genomes was performed by developing a semi-automatic analysis pipeline. The SGP2 gene-finding tool was developed from procedures used in this pipeline. The concept behind SGP2 is that similarity regions obtained by TBLASTX are used to increase the score of exons predicted by geneid, in order to produce a more accurate set of gene structures. SGP2 provides a specificity that is high enough for its predictions to be experimentally verified by RT-PCR. The RT-PCR validation of predicted splice junctions also serves as example of how combined computational and experimental approaches will yield the best results. Then, we performed a descriptive analysis at sequence level of the splice site signals from a reliable set of orthologous genes for human, mouse, rat and chicken. We have explored the differences at nucleotide sequence level between U2 and U12 for the set of orthologous introns derived from those genes. We found that orthologous splice signals between human and rodents and within rodents are more conserved than unrelated splice sites. However, additional conservation can be explained mostly by background intron conservation. Additional conservation over background is detectable in orthologous mammalian and chicken splice sites. Our results also indicate that the U2 and U12 intron classes have evolved independently since the split of mammals and birds. We found neither convincing case of interconversion between these two classes in our sets of orthologous introns, nor any single case of switching between AT-AC and GT-AG subtypes within U12 introns. In contrast, switching between GT-AG and GC-AG U2 subtypes does not appear to be unusual. Finally, we implemented visualization tools to integrate annotation features for gene- finding and comparative analyses. One of those tools, gff2ps, was used to draw the whole genome maps for human, fruitfly and mosquito. gff2aplot and the accompanying parsers facilitate the task of integrating sequence annotations with the output of homologybased tools, like BLAST.We have also adapted the concept of pictograms to the comparative analysis of orthologous splice sites, by developing compi.

Η παρούσα διατριβή ανήκει στο γενικότερο θεματικό πεδίο της σεισμικής αποτίμησης, σχεδιασμού ή ανασχεδιασμού κατασκευών οπλισμένου σκυροδέματος με βάση τις μετακινήσεις. Οι σύγχρονες μέθοδοι αυτού του τύπου, στηρίζονται σε έλεγχο και σύγκριση της σεισμικής απαίτησης με την ικανότητα των μελών της κατασκευής σε όρους μετακινήσεων παρά σε όρους δυνάμεων. Δημιουργείται επομένως η ανάγκη για απλό και αξιόπιστο υπολογισμό της συμπεριφοράς μελών οπλισμένου σκυροδέματος σε κάμψη και διάτμηση, σε όρους μετακινήσεων. Το αντικείμενο της παρούσης διατριβής είναι η ανάπτυξη προσομοιωμάτων για τον υπολογισμό των βασικών χαρακτηριστικών της συμπεριφοράς καμπτόμενων μελών οπλισμένου σκυροδέματος και συγκεκριμένα: της ροπής διαρροής, της παραμόρφωσης στη διαρροή, της ενεργού δυσκαμψίας, της παραμόρφωσης στην αστοχία, της διατμητικής αντοχής σε ανακυκλιζόμενη φόρτιση, της αντοχής μελών με χαμηλό λόγο διάτμησης και της συμπεριφοράς υπό διαξονική καταπόνηση. Εξετάζονται μέλη διαφόρων τύπων και διαφορετικής διατομής, μέλη με ενίσχυση μανδύα οπλισμένου σκυροδέματος ή μανδύα σύνθετων υλικών, καθώς επίσης και μέλη με μάτιση του διαμήκους οπλισμού στην περιοχή πλαστικής άρθρωσης. Για την ανάπτυξη των προσομοιωμάτων, καθώς και για τον έλεγχο άλλων παλαιότερων, αναπτύχθηκε και αξιοποιήθηκε βάση πειραματικών δεδομένων μελών οπλισμένου σκυροδέματος με περισσότερα από 2800 πειράματα από τη διεθνή βιβλιογραφία. Για τον υπολογισμό της ροπής και της καμπυλότητας στη διαρροή, αναπτύσσονται απλές σχέσεις υπολογισμού, βασιζόμενες σε ανάλυση σε επίπεδο διατομής και καθορίζονται τα κατάλληλα κριτήρια διαρροής. Αναπτύσσονται ακολούθως σχέσεις υπολογισμού της παραμόρφωσης στη διαρροή, και συγκεκριμένα της γωνίας στροφής χορδής του μέλους στη διαρροή, θy, ως άθροισμα τριών όρων: καμπτικής παραμόρφωσης, διατμητικής παραμόρφωσης και παραμόρφωσης λόγω ολίσθησης των ράβδων διαμήκους οπλισμού από την περιοχή αγκύρωσης. Προτείνονται δε δύο εναλλακτικοί τρόποι υπολογισμού της ενεργού δυσκαμψίας, ένας θεωρητικός και ένας καθαρά εμπειρικός. Στη συνέχεια εξετάζεται η παραμόρφωση στην αστοχία και προτείνονται δύο εναλλακτικοί μέθοδοι υπολογισμού της γωνίας στροφής χορδής στην αστοχία, θu. Η 1η βασίζεται στον υπολογισμό της καμπυλότητας στην αστοχία, φu, με εφαρμογή του κατάλληλου προσομοιώματος περίσφιγξης του σκυροδέματος, και στην εφαρμογή της φu σε μήκος πλαστικής άρθρωσης ίσο με Lpl, ενώ η 2η σε καθαρά εμπειρικές εξισώσεις. Εξετάζεται ακολούθως η διατμητική αντοχή σε ανακυκλιζόμενη φόρτιση και προτείνονται προσομοιώματα για αστοχία σε διαγώνιο εφελκυσμό ή αστοχία σε λοξή θλίψη, μετά την καμπτική διαρροή. Στη συνέχεια εξετάζεται η συμπεριφορά μελών οπλισμένου σκυροδέματος υπό διαξονική καταπόνηση. Εξετάζονται επίσης μέλη με χαμηλό λόγο διάτμησης και προτείνονται νέα αντιπροσωπευτικότερα κριτήρια για τον χαρακτηρισμό ενός μέλους ως “κοντό μέλος”, καθώς και νέα μεθοδολογία υπολογισμού της αντοχής των μελών αυτών, με κατάλληλο συνδυασμό του προσομοιώματος των Shohara and Kato, 1981 και των Φαρδής και συνεργάτες 1998. Ακολούθως εξετάζονται μέλη ενισχυμένα με μανδύα σύνθετων υλικών και προτείνονται προσομοιώματα υπολογισμού της γωνίας στροφής χορδής στη διαρροή και την καμπτική αστοχία, καθώς και προσομοίωμα υπολογισμού της διατμητικής αντοχής. Στη συνέχεια εξετάζεται η συμπεριφορά μελών με μάτιση του διαμήκους οπλισμού στην περιοχή πλαστικής άρθρωσης, καθώς και η εφαρμογή μανδύα σύνθετων υλικών για την ενίσχυση της περιοχής αυτής. Τέλος εξετάζεται η συμπεριφορά στη διαρροή και στην αστοχία, μελών ενισχυμένων με μανδύα οπλισμένου σκυροδέματος. Η ανάπτυξη όλων των προτεινόμενων προσομοιωμάτων της διατριβής βασίζεται στην καλύτερη δυνατή συμφωνία με τα πειραματικά αποτελέσματα της βάσης δεδομένων, χωρίς όμως να θυσιάζεται η απλότητα και η ευχρηστία αυτών. / The present Thesis belongs in the general field of seismic assessment, design and redesign of concrete structures with displacement based procedures. Modern methods of this kind are based in controlling and comparing seismic demand with structural elements capacity in terms of displacements rather than forces. This leads in the need of estimating reinforced concrete elements performance under bending and shear, in terms of displacements. The object of the Thesis is development of models for calculating the basic performance characteristics of reinforced concrete elements under bending, in particular: yield moment, deformation at yielding, effective stiffness, deformation at ultimate, shear strength under cyclic loading, maximum strength of members with low shear ratio and behavior under biaxial loading. Members with various types of section and various characteristics are included, as also members retrofitted with FRP jacket or concrete jacket and members with lap-splice of longitudinal reinforcement in plastic hinge region. In order to develop new models and check older ones, a database of more than 2800 experiments from international literature on reinforced concrete elements was created and used here. Simple equations and procedures are suggested for calculating yield moment and corresponding curvature, based on section analysis, by specifying the appropriate yield criteria. Equations for calculating deformation at yielding, in particular chord rotation at yielding, θy as the sum of deformations due to bending, due to shear and due to slippage of longitudinal reinforcement from anchorage zone, are also developed. Calculation of effective stiffness is based on two alternative models, one theoretical and one purely empirical. Deformation at ultimate is then examined where two methods for calculating chord rotation at ultimate are suggested. 1st one is based on ultimate curvature, φu, where an appropriate concrete confinement model is used, and plastic hinge length Lpl, while 2nd one is based on purely empirical equations. Shear strength under cyclic loading is also examined and new models for calculating shear strength for shear tension and shear compression failure after flexural yield are developed. Behavior of reinforced concrete elements under biaxial loading is then examined. Elements with low shear ratio are also covered and new, more representative, criteria to characterize an element as a “short element” are suggested. A procedure based on an appropriate combination of Shohara and Kato 1981 model and Fardis et al. 1998 model is then suggested for calculating maximum strength of such “short elements”. Retrofitted members with FRP jacket are then examined and models for chord rotation at yielding and ultimate, as well as for shear strength are suggested. Behavior of members with lap-splice of longitudinal reinforcement inside plastic hinge region is then examined, including also retrofitting of this region with FRP jacket. Performance at yielding and ultimate of retrofitted members with concrete jacket is also examined. Development of all the suggested models of the Thesis is based on best fit with experimental results of the database, without sacrificing simplicity and applicability of the models.

No description available.

No description available.

Human α-N-Acetylglucosaminidase (Naglu) is a lysosomal acid hydrolase implicated in tthe rare metabolic storage disorder known as mucopolysaccharidosis type IIIB (MPS IIIB; also Sanfilippo syndrome B). Absence of this enzyme results in cytotoxic accumulation of heparan sulphate in the central nervous system, causing mental retardation and a shortened lifespan. Enzyme replacement therapy is not currently effective to treat neurological symptoms due to the inability of exogenous Naglu to access the brain. This laboratory uses a Spodoptera frugiperda (Sf9) insect cell system to express Naglu fused to a synthetic protein transduction domain with the intent to facilitate delivery of Naglu across the blood-brain barrier. The project described herein may be broken down into three main sections. Firstly, the impact of two cryptic splice sites on Naglu expression levels was analyzed in both transiently expressing Sf9 cultures and stably selected cell lines. Secondly, the effectiveness of the native Naglu secretion-signaling peptide in the Sf9 system was examined. Finally, purification of a Naglu fusion protein from suspension culture medium was performed using hydrophobic interaction chromatographic techniques. The ultimate goal of this research is to develop an efficient system for economical, large-scale production of a human recombinant Naglu fusion protein that has the potential to be successfully used for enzyme replacement therapy to treat MPS IIIB. / Graduate