In the human genome, 43 BTB (Bric-à-brac, Tramtrack, and Broad Complex) containing BTB-Zinc Finger proteins have been identified, many of which are transcription factors involved in cancer and development. These BTB domains have been shown to form homodimers and heterodimers which raise DNA binding affinity and specificity for transcription factors. This project was to develop an efficient assay to systematically identify interactions between BTB domains. It combined a co-expression system, fluorescent protein tagging and Ni-NTA plate retention. It was concluded that fourteen analyzed BTB domains formed homodimers, but only certain BTB pairs formed heterodimers, such as BCL6 with Miz1 and Miz1 with RP58. To further understand the specificity of BTB domain interactions, more structural and sequence information is still needed. In conclusion, this assay provided a comprehensive detection method for BTB domain interaction mapping. The information generated provides candidates for further functional and structural studies.

A low-cost power router (PR), capable of dynamic, independent control of active- and reactive-power flows on meshed grids is presented. The operating principle, detailed schematics, and various possible implementations of the proposed power router are discussed. Various operating modes are identified and a control algorithm has been proposed and verified through simulations. Small-signal and frequency-domain models of the power router from basic time-domain equations are developed. A three-tier protection system based on the fail-normal switch to avoid single point-of-failure is presented. The operation of proposed protection system in isolating the converter and the grid in the event of faults is verified through simulation. An analytical method to evaluate the stability of a system with multiple power routers is proposed. Necessary conditions for the PR-controller design to ensure stable operation of a system with multiple power routers is proposed. These necessary conditions are verified through simulation studies. Potential applications of proposed power router in distribution system and the associated challenges in implementation are presented. The functionality and advantages of the proposed power router are experimentally demonstrated at 13 kV, 1 MVA. The proposed power router can result in a low cost power routing solution that can reduce electric grid congestion and efficient implementation of RPS mandates.

Differential gene expression plays a key role in developmental pathways within an organism. Examples of such pathways include primary sex determination signaling and the formation of secondary sexual characteristics. This dissertation is focused on the use of suppression subtractive hybridization (SSH) to identify genes that are differentially expressed and involved in some aspect of sexual development in the Mediterranean fruit fly (medfly), Ceratitis capitata. In the course ofthis project, a method for sexing individual specimens from pre-adult stages was developed. This method was used to collect sex-specific RNAs at different developmental stages for use in SSH. A total of25 subtraction products were obtained across all the stages examined. Analysis of these products revealed that approximately half were similar to cytoplasmic ribosomal proteins and mitochondrial ribosomal RNA The remaining products represent putative medfly homologs of other previously identified genes or potentially novel genes One ofthe subtraction products, representing a potentially novel gene, was characterized in detail. This gene, named mapotge', represents a novel medfly gene that appears to encode a polypeptide of 299 amino acids. The N-terminus of this polypeptide contains a BTB-POZ domain. This domain functions as a protein-protein interaction motif found in a wide range of organisms from humans to Drosophila that mediates protein dimerization and oligomerization. The temporal expression pattern of mapotge' was determined using RT-PCR and Northern blot analysis. These revealed that the transcript is expressed throughout embryogenesis in both females and males, and in adult females that are > 0.5 days post-eclosion. Minimal expression is observed in female and male third instar larvae, early pupae, and in adult males. Studies were also initiated to characterize the representation of additioual sequences containing a BTB-POZ domain in the medfly genome. This was performed using Southern blot analysis and degenerate primers for the polymerase chain reaction (PCR). These results indicate the presence of at least three sequences in the medfly, in addition to 'mapotge', that contain a BTB-POZ domain. Potential evolutionary relationships ofthe BTB-POZ domain sequences from the medfly and other insect species were also analyzed.

Bovine tuberculosis (BTB) is a zoonotic disease that affects domestic and wild animals, and humans. It is caused by Mycobacterium bovis (M. bovis) and has a wide host range. The effective control of BTB is of paramount importance and this can be achieved through the use of accurate and comprehensive diagnostic tests. The most widely used methods to detect BTB are the skin test and in vitro gamma interferon assay which do not detect anergic animals, but serological tests such as ELISA and fluorescence polarization assay (FPA) have been found promising in ancilliary tuberculosis diagnosis. The overall aim was to study M. tuberculosis complex (MTBC) protein, mycobacterial protein bovis 70 (MPB70) as a target for serological assays in the detection of antibodies to bovine tuberculosis. The MPB70 protein was expressed, purified and labeled with fluorescein (FITC). The mpb70 gene was fragmented into three regions without disrupting predicted epitopes. The resulting protein Fragments were expressed as fusion proteins with the monster green fluorescent protein (MGFP). The recombinant MPB70 (rMPB70) and the expressed gene fragments 2&3 were tested in immunoblots and ELISAs. The rMPB70 and fragment 2-MGFP reacted with chicken antibodies raised against rMPB70 and immune sera from BTB infected buffaloes. MPB70 peptides were synthesized as an approach to identify even smaller antigenic regions. The peptides BT1G (residues 31-45) and BT51L (residues 81-95) were recognised by anti-MPB70 chicken antibodies in the ELISA and fall within fragment 1 and 2, respectively. The tracers (rMPB70-FITC, fragment 2-MGFP fusion and peptides BT1G&BT51L) were tested in the FPA, but the results failed to distinguish between immune sera from chickens immunized with rMPB70 and negative control sera. Even though the FPA was not successful, the MPB70 fragment 2-MGFP fusion protein, which was recognized by sera from BTB infected buffaloes, was tested in an ELISA using panels of sera from uninfected and naturally M. bovis infected buffaloes and cattle. The diagnostic performance of the ELISA was, however, overall unsatisfactory and hence of very limited use as a serological test to detect antibody responses to BTB as a stand-alone assay. Sera from some of the animals gave false positive reactions indicating that MPB70 was not sufficiently specific for serodiagnosis of M. tuberculosis complex infections. / Dissertation (MSc)--University of Pretoria, 2012. / Veterinary Tropical Diseases / unrestricted

Generation and characterization of activation tagged potato mutants could aid in functional genomic studies. Morphological and molecular studies were conducted to compare potato cv. Bintje, its two mutants, underperformer (up), and nikku generated using the activation tagging vector pSKI074, and nikku revertant plants. Mutant up exhibited a dwarf phenotype (plant height 42 cm vs. 73 cm in cv. Bintje), abundant axillary shoot growth (3.1 shoots/plant compared to 0.7 shoots/plant in cv. Bintje; in vitro plants), greater tuber yield, altered tuber traits and early senescence compared to wild-type Bintje under in vitro conditions. Under in vivo conditions, the dwarf and early senescence phenotypes of the mutant were consistent, but the tuber yield of up was less (250 g/plant compared to 610 g/plant in wild-type Bintje) and had fewer axillary shoots compared to wild-type (1.9 shoots/plant in up vs. 4.7 shoots/plant in Bintje). Mutant nikku plants exhibited an extremely dwarf phenotype (plant height 2 cm in nikku vs. 6 cm in Bintje), had small hyponastic leaves, were rootless, and infrequently produced small tubers when compared to cv. Bintje. The overall nikku phenotype was suggestive of a constitutive stress response, which was further supported by the higher expression levels of several stress-responsive genes in nikku. The nikku revertant plants exhibited near normal stem elongation, larger leaves and consistent rooting, and it was a case of partial reversion. Southern blot analyses indicated the presence of single T-DNA insertions on chromosome 10 in the up and on chromosome 12 in the nikku mutant. The reversion in the nikku plants was not associated with the loss of enhancer copies from the original nikku mutant. Reverse transcriptase PCR analyses indicated transcriptional activation/repression of several genes in the up and nikku mutants, suggesting pleiotropic effects. In revertant, the expression levels of several genes which were differentially regulated in the nikku mutant were similar to Bintje. The gene immediately flanking the right border of the T-DNA insertion, which encoded a novel BTB/POZ (Broad complex, Tramtrac, Bric a brac; also known as Pox virus and Zinc finger) domain-containing protein, was highly up-regulated in the up mutant. This protein domain plays an important role in several important developmental, transcriptional and regulatory pathways. The mRNA-seq analyses resulted in 1,632 genes that were differentially expressed between mutant up and Bintje and the total number of up-regulated genes (661) were less than the number of genes down-regulated (971 genes) in the up mutant. Further analyses indicated that a variety of biological processes including decreased cell division, cell cycle activity, and abiotic stress responses were modified in the up mutant. In the nikku mutant, two potato genes, encoding an Acyl-CoA N-acyltransferases (NAT) superfamily protein, and a predicted major facilitator superfamily protein (MFS) were identified and overexpression lines Bintje/35S::NAT1 and Bintje/35S::PMT1 were created for recapitulation of the nikku mutant phenotype. Methylated DNA-PCR between the nikku and the revertant indicated a change in methylation status of the 35S enhancers, suggesting that the nikku revertant phenotype may be associated with some epigenetic modification. / Ph. D.

在工業、農業、或醫藥界的實驗中，經常必須拿數個不同的試驗處理 (test treatments)和一個已使用過的對照處理(control treatment)比較 。所謂的試驗處理可能是數組新的儀器、不同配方的新藥、或不同成份的 肥料等。以實驗新藥為例，研藥者想決定是否能以新藥取代原來所使用的 藥，故對v種新藥與原藥做比較，評估其藥效之差異。為了降低實驗中不 必要的誤差以增加其準確性，集區設計成為實驗者常用的設計方法之一； 又因A式最適設計是我們欲估計的對照處理效果(effect)與試驗處理效果 之差異之估計值最小的設計，基於此良好的統計特性，我們選擇A式最適 性為評判根據。古典的A式最適性並未將對照處理與試驗處理所具備的先 前資訊(prior information)加以考慮，以上例而言，我們不可能對原來 使用的藥一無所知，經由過去的實驗或臨床的反應，研藥者必已對其藥性 有某種程度的了解，直觀上，這種過去經驗的累積，影響到實驗配置上， 可能使對照處理的實驗次數減少，相對地可對試驗處理多做實驗，設計遂 更具意義。因而本文考慮在k>v的情形下之貝式最適集區設計，對先前分 配施以某種限制，依據準確設計理論(exact design theory)，推導單項 異種消除模型(one- way elimination of heterogeneity model)之下的 貝氏A式最適設計與Γ- minimax最適設計，使Majumdar(1992)的結果能適 用於完全集區設計。此種設計對先前分配具有強韌性，即當先前分配有所 偏誤，且其誤差在某一範圍內時，此設計仍為最適設計或仍可維持所謂的 高效度(high efficiency)。本文將列舉許多實例以說明此一特性。 We consider the problem of comparing a set of v test treatments simultaneously with a control treatment when k>v. Following the work of Majumdar(1992), we use exact design theory to derive Bayes A-optimal designs and optimal Γ-minimax designs for the one-way elimination of heterogeneity model. These designs have the same properties as of Bayes A-optimal incomplete block designs. We also provide several examples of robust optimal designs and highly efficient designs.

The thesis dealt with newly built detached house for seniors. The building is designed in the village Drnovice, Blansko district. House for seniors has one basement and three floors. In the basement are designed therapeutic ingredients such as physiotherapy, massage, gym and the appropriate sanitary facilities. Further there are technical facilities, sanitary facilities and lockers for employees. The first floor is designed to entry, reception, library, lounge, dining room and other spaces. Second and third floor is designed for living. Dimensions of the maximum contour of the house are 55,14 x30,34 m. Foundations are of strip footings of plain concrete C16/20. The thickness of the base concrete slab is 150mm and is reinforcer with KARI networks 100/100/6. The building is made of Heluz system, perimeter basement walls is made of the BTB fittings. The staircase is a reinforced concrete slab. The roof of the house is flat, single.

A la suite d’un criblage in silico nous avons identifié un nouveau gène codant pour une protéine à domaine BTB-POZ, XBTBD6.<p>Nous avons déterminé que la protéine XBTBD6 est une protéine cytoplasmique. Dans les cellules Hela, CHO, U2OS et COS7 la protéine XBTBD6 est localisée dans des corpuscules cytoplasmiques, localisation similaire à celle des protéines XBTBD3, HBTBD1 et HBTBD2. Nous avons observé que la partie N-terminale de la protéine, contenant le domaine BTB-POZ, est localisée dans la cellule comme la protéine entière ;par contre la partie C-terminale est exclusivement nucléaire. De plus, nous avons observé que XBTBD6 est localisée de façon diffuse dans le cytoplasme des cellules Neuro2A, 9L et 518A2e. Nous avons montré que la protéine XBTBD6 homodimérise et hétérodimérise avec XBTBD3 et XBTBD2 et qu’elle interagit avec l’ubiquitine ligase E3 XCullin 3. L’ensemble de ces interactions nécessite la présence du domaine BTB-POZ. Ces données montrent que les protéines BTBD6, BTBD3, BTBD1 et BTBD2 possèdent des propriétés communes indiquant qu’elles appartiennent à un sous groupe de la famille des protéines à domaine BTB-POZ.<p>Le profil d’expression a été analysé par la technique de protection à la RNAse et par hybridation in situ. Les résultats montrent que ce gène est fortement exprimé dans le système nerveux adulte et embryonnaire. Des expériences de surexpression par micro-injection d’ARNm ont permis de placer le gène XBTBD6 dans la cascade d’activation des gènes proneuraux en aval de XNgnr-1, XNeuroD, Xath3 et Xebf3. Ces résultats montrent que XBTBD6 est un marqueur neuronal chez le xénope. <p>Au cours de l’étude de la fonction du gène XBTBD6, nous avons montré que la surexpression et la perte de fonction de ce gène dans l’embryon de xénope n’induit pas de variation du nombre de neurones dans la plaque neurale. Par contre nous avons observé que la surexpression du gène XBTBD6 dans des cellules Neuro2A en différentiation régule négativement la croissance des neurites.<p>Nous avons élaboré un modèle de fonctionnement biochimique hypothétique où la protéine XBTBD6 fonctionnerait comme protéine adaptatrice dans un complexe d’ubiquitination permettant l’ubiquitination d’une protéine cible. Nous avons recherché les partenaires potentiels de XBTBD6 en utilisant la technique du double hybride en levure mais sans y parvenir.<p> / Doctorat en sciences, Spécialisation biologie moléculaire / info:eu-repo/semantics/nonPublished

Power flow (PF) control can increase the utilization of the transmission system and connect lower cost generation with load. While PF controllers have demonstrated the ability to realize dynamic PF control for more than 25 years, PF control has been sparsely implemented. This research re-examines PF control in light of the recent development of fractionally-rated PF controllers and the incremental power flow (IPF) control concept. IPF control is the transfer of an incremental quantity of power from a specified source bus to specified destination bus along a specified path without influencing power flows on circuits outside of the path. The objectives of the research are to develop power system operation and planning methods compatible with IPF control, test the technical viability of IPF control, develop transmission planning frameworks leveraging PF and IPF control, develop power system operation and planning tools compatible with PF control, and quantify the impacts of PF and IPF control on multi-decade transmission planning. The results suggest that planning and operation of the power system are feasible with PF controllers and may lead to cost savings. The proposed planning frameworks may incent transmission investment and be compatible with the existing transmission planning process. If the results of the planning tool demonstration scale to the national level, the annual savings in electricity expenditures would be $13 billion per year (2010$). The proposed incremental packetized energy concept may facilitate a reduction in the environmental impact of energy consumption and lead to additional cost savings.