Thesis (Ph. D.)--Virginia Polytechnic Institute and State University, 1993. / Vita. Abstract. Includes bibliographical references (leaves 109-117). Also available via the Internet.

Thesis (Ph. D.)--Ohio University, August, 2004. / Includes bibliographical references (p. 185-196).

Thesis (doctoral)--Universität Regensburg, 2009. / Includes bibliographical references.

Thesis (Ph.D.)--Ohio University, August, 2004. / Title from PDF t.p. Includes bibliographical references (p. 185-196)

Thesis (Ph. D.)--Illinois State University, 1996. / Title from title page screen, viewed May 25, 2006. Dissertation Committee: Charles F. Thompson, Angelo P. Capparella (co-chairs), Steven A. Juliano, Anthony J. Otsuka, Scott K. Sakaluk, David F. Weber. Includes bibliographical references (leaves 78-84) and abstract. Also available in print.

Thesis (Ph. D.)--University of Hong Kong, 2007. / Also available online.

Thesis (MSc)--University of Stellenbosch, 2000. / ENGLISH ABSTRACT: Streptomyces spp. are responsible for a large proportion of the world-wide quality deterioration of potatoes causing a potato tuber disease called cornmon scab. Determining the genetic diversity of the Streptomyces spp., especially the main pathogen, S. scabies, has been a prerequisite for the ultimate control of common scab. Techniques responsible for the classification and determination of genetic diversity have improved with advances in DNA technology. Analysis of South African (S.A.) S. scabies isolates has been focusing on the organisms' morphology, physiology, pathogenicity and melanin production, but the classification of S. scabies using DNA techniques has not yet been explored. In this study various DNA techniques were screened for optimal use in determining the genetic diversity within and among isolates of S. scabies. Bacteria had been sampled from the main potato producing regions in S.A. and a few other regions. The techniques explored included RAPDs, AFLPs, RAMS, Rep-PCR, 16S rDNA sequencing and ITS analysis. The first three techniques had to be abandoned due to non-reproducibility between the same isolate extracted on separate occasions and ITS analysis was abandoned due to sequencing difficulties. Of the three Rep-PCR techniques tested (BOX, ERIC and REP), BOX was selected because it produced the clearest and most reproducible results. BOX-PCR and 16S rDNA sequencing were therefore ultimately selected as the methods to analyse the genetic diversity of the S. scabies isolates. Information concerning the pathogenicity of the isolates was supplied by the Vegetable and Ornamental Plant Research Institute of the Agricultural Research Council (VOPI, ARC, Roodeplaat). A brief analysis of the pathogenicity prediction of the isolates in this study was explored with the PCR technique. Presence of the necJ gene was previously shown to be an indication of the pathogenicity within the Streptomyces spp. group. PCR analysis is based on the amplification of a O.72kb fragment (necl) in pathogenic isolates which was absent in non-pathogenic isolates. However, in this study the test for pathogenicity lacked specificity and sensitivity and some of the problems experienced included non-reproducibility between PCR reactions and the presence of the pathogenic fragment in the nonpathogenic isolates (as designated by VOPI, ARC). These observations led to the conclusion that this technique is not an ultimate test for pathogenicity of S. scabies isolates in a South African context. The genetic distances and similarity matrices of the Rep-PCR results were calculated using Nei's genetic distance calculation (Nei M, 1975). Clusters from these matrices were constructed using the unweighted pair group average (UPGMA) with the PAUP4 package. The clusters for the 16S rDNA sequences were formed with the Neighbor Joining (NJ) method and the PAUP4 package. The NJ trees do not take small sequencing differences into account, therefore a Parsimony Network had to be constructed. The trees obtained with the 16S rDNA sequencing techniques grouped most S. scabies isolates into one major group with a 100% bootstrap robustness of this group. More genetic diversity was illustrated by the BOX-PCR technique and the isolates were generally grouped according to their different regions of origin. However, the bootstrap values were low, indicating a lack of robustness regarding the BOXPCR clustering. This was not unexpected as the number of data points employed in the BOX technique is very limited. Both techniques revealed unexpected grouping of a few isolates. Their isolated positions could be attributed to possible misclassification or to the fact that they could be genetically different S. scabies isolates. Streptomyces spp. (other than S. scabies) displayed enough differences to place them in their own distinct groups using both techniques. Comparison of the cluster results obtained in this study did not correlate to the data supplied by the VOPI, ARC (morphology, physiology, pathogenicity and melanin production) which revealed differences between the S. scabies isolates within their respective regions. The lack of diversity displayed by the 16S rDNA technique can be attributed to the fact that only a limited section of the genome is involved making it inappropriate for intra-species genetic diversity analysis. The BOX technique takes various loci within the genome but is still not ideal for a thorough genetic diversity analysis. This study represents the first attempt to determine the genetic diversity of S. scabies in S.A. on DNA level. / AFRIKAANSE OPSOMMING: Streptomyces spp. is verantwoordelik vir 'n _groot deel van die wereld afname in aartappel kwaliteit as gevolg van die aartappelknol siekte bruinskurf. Die bepaling van die genetiese diversiteit tussen die Streptomyces spp., varal die hoof patogeen in die groep, S. scabies, is 'n vooreiste vir die uiteindelike beheer van bruinskurf. Tegnieke verantwoordelik vir die klassifikasie en bepaling van genetiese diversiteit het verbeter met vooruitgang in DNA tegnologie. Analise van Suid Afrika (S.A.) se S. scabies isolate konsentreer op die organisme se morfologie, fisiologie, patogenisiteit en malanien produksie, maar die klassifikasie van S. scabies met die behulp van DNA tegnieke is nog nie uitgevoer me. In hierdie studie is verskeie DNA tegnieke ondersoek vir optimale bepaling van genetiese diversiteit binne en tussen S. scabies isolate van S.A Bakteriee is verkry van die hoof aartappel-produserende areas in S.A. en ook van 'n paar ander areas. Die tegnieke wat in die studie gebruik is, het RAPDs, AFLPs, Rep-PKR, 16S rDNA volgordebepaling en ITS analise ingesluit. Die eersgenoemde drie tegnieke is uitgesluit as gevolg van nie-herhalende resultate tussen dieselfde isolaat geisoleer op verskillende geleenthede. ITS analise is uitgesluit as gevolg van probleme met volgordebepaling. Rep- PKR en 16S rDNA volgordebepaling is uiteindelik gekies as die mees geskikte metodes vir die analise van genetiese diversiteit tussen S. scabies isolate in hierdie studie omdat albei skynbare herhaalbare resultate gelewer het. Inligting met betrekking tot die patogenisiteit van die isolate is voorsien deur die Groente en Sierplant Instituut van die Landbou Navorsinsraad (YOPI, LNR). 'n Vinnige analise van die patogenisiteits voorspelling van die isolate is uitgevoer met die PKR tegniek. Dit is voorheen aangetoon dat die teenwoordiheid van die necJ geen dui op die patogenisiteit in die Streptomyces sp _groep. PKR analise het 'n O.72kb fragment (necl) in patogeniese isolate geamplifiseer wat nie teenwoordig was in niepatogeniese isolate nie. Hierdie toets vir patogenisiteit soos gebruik in hierdie studie was egter onspesifiek en onsensitief en sommige van die probleme wat ondervind is sluit in nie-herhaalbaarheid tussen PKR reaksies en die teenwoordigheid van die patogeniese fragment in die nie-patogeniese isolaat (soos beskryf deur YOPI, LNR). Uit hierdie waarnemings word afgelei dat die tegniek nie 'n geskikte toets vir patogenisiteit van S. scabies isolate in 'n S.A konteks is nie. Die genetiese afstande en ooreenkomstige matrikse van die Rep-PCR resultate is bereken met die genetiese afstand bepaling van Nei (Nei M, 1975). Groepe is gevorm met die "unweighted pair group average" (UPGMA) en PAUP4 pakket. Die "Neighbor Joining" (NJ) groepe is gevorm met die 16S rDNA volgordebepaling data mbv die PAUP4 pakket. Die NJ groepering neem nie klein volgorde verskille in ag nie en gevolglik moes'n "Parsimony Network" opgestel word. Die groepering met die 16S rDNA volgordebepaling het meeste van die isolate in een hoof groep geplaas met 'n 100% "bootstrap" waarde. Meer genetiese diversiteit is met die BOX-PCR tegniek gevind en isolate was oor die algemeen gegroepeer vol gens van hul oorsprong. Die "bootstrap" waardes vir die BOX tegniek was baie laag. Dit was nie onverwags nie, want die hoeveelheid data punte was beperk met die BOX tegniek. Albei tegnieke het 'n aantal afwykende isolate vertoon. Hul ge-isoleerde posisies kan toegeskryf word aan moontlike misklassifikasies van die isolaat. Die moontlikheid dat daar wel genetiese verkille tussen die isolate is, kan egter nie uitgesluit word nie. Streptomyces spp. (uitgesluit S. scabies) het genoeg variasie vertoon om hulle in hul eie groepe met die gebruik van beide tegnieke te plaas. Vergelyking tussen die groepe in die studie stem nie ooreen met die data verkry vanaf YOPl, LNR (morfologie, fisiologie, patogenesiteit en melanien produksie) nie wat verskille tussen S. scabies isolate binne 'n sekere gebied vertoon. Die gebrek aan diversiteit soos vertoon deur die 16S rDNA tegniek kan toegeskryf word aan die feit dat slegs 'n beperkte gedeelte van die genoom ondersoek word, wat dit ongeskik vir intra-species genetiese diversiteit analise maak. Die BOX tegniek neem verskeie loci in die genoom in ag, maar is steeds nie ideaal vir deeglike genetiese diversiteit analise nie. Hierdie studie verteenwoordig die eerste poging om die genetiese diversiteit van S. scabies in S.A. op DNA vlak te bepaal.

RF fingerprinting can solve the indoor positioning problem with satisfactory accuracy, but the methodology depends on the so-called radio map calibrated in the offline phase via manual site-survey, which is costly, time-consuming and somewhat error-prone. It also assumes the RF fingerprint’s signal-spatial correlations to remain static throughout the online positioning phase, which generally does not hold in practice. This is because indoor environments constantly experience dynamic changes, causing the radio signal strengths to fluctuate over time, which weakens the signal-spatial correlations of the RF fingerprints. State-of-the-arts have proposed adaptive RF fingerprint methodology capable of calibrating the radio map in real-time and on-demand to address these drawbacks. However, existing implementations are highly server-centric, which is less robust, does not scale well, and not privacy-friendly. This thesis aims to address these drawbacks by exploring the feasibility of implementing an adaptive RF fingerprint indoor positioning system in a distributed and client-centric architecture using only commodity Wi-Fi hardware, so it can seamlessly integrate with existing Wi-Fi network and allow it to offer both networking and positioning services. Such approach has not been explored in previous works, which forms the basis of this thesis’ main contribution. The proposed methodology utilizes a network of distributed location beacons as its reference infrastructure; hence the system is more robust since it does not have any single point-of-failure. Each location beacon periodically broadcasts its coordinate to announce its presence in the area, plus coefficients that model its real-time RSS distribution around the transmitting antenna. These coefficients are constantly self-calibrated by the location beacon using empirical RSS measurements obtained from neighbouring location beacons in a collaborative fashion, and fitting the values using path loss with log-normal shadowing model as a function of inter-beacon distances while minimizing the error in a least-squared sense. By self-modelling its RSS distribution in real-time, the location beacon becomes aware of its dynamically fluctuating signal levels caused by physical, environmental and temporal characteristics of the indoor environment. The implementation of this self-modelling feature on commodity Wi-Fi hardware is another original contribution of this thesis. Location discovery is managed locally by the clients, which means the proposed system can support unlimited number of client devices simultaneously while also protect user’s privacy because no information is shared with external parties. It starts by listening for beacon frames broadcasted by nearby location beacons and measuring their RSS values to establish the RF fingerprint of the unknown point. Next, it simulates the reference RF fingerprints of predetermined points inside the target area, effectively calibrating the site’s radio map, by computing the RSS values of all detected location beacons using their respective coordinates and path loss coefficients embedded inside the received beacon frames. Note that the coefficients model the real-time RSS distribution of each location beacon around its transmitting antenna; hence, the radio map is able to adapt itself to the dynamic fluctuations of the radio signal to maintain its signal-spatial correlations. The final step is to search the radio map to find the reference RF fingerprint that most closely resembles the unknown sample, where its coordinate is returned as the location result. One positioning approach would be to first construct a full radio map by computing the RSS of all detected location beacons at all predetermined calibration points, then followed by an exhaustive search over all reference RF fingerprints to find the best match. Generally, RF fingerprint algorithm performs better with higher number of calibration points per unit area since more locations can be classified, while extra RSS components can help to better distinguish between nearby calibration points. However, to calibrate and search many RF fingerprints will incur substantial computing costs, which is unsuitable for power and resource limited client devices. To address this challenge, this thesis introduces a novel algorithm suitable for client-centric positioning as another contribution. Given an unknown RF fingerprint to solve for location, the proposed algorithm first sorts the RSS in descending order. It then iterates over this list, first selecting the location beacon with the strongest RSS because this implies the unknown location is closest to the said location beacon. Next, it computes the beacon’s RSS using its path loss coefficients and coordinate information one calibration point at a time while simultaneously compares the result with the measured value. If they are similar, the algorithm keeps this location for subsequent processing; else it is removed because distant points relative to the unknown location would exhibit vastly different RSS values due to the different site-specific obstructions encountered by the radio signal propagation. The algorithm repeats the process by selecting the next strongest location beacon, but this time it only computes its RSS for those points identified in the previous iteration. After the last iteration completes, the average coordinate of remaining calibration points is returned as the location result. Matlab simulation shows the proposed algorithm only takes about half of the time to produce a location estimate with similar positioning accuracy compared to conventional algorithm that does a full radio map calibration and exhaustive RF fingerprint search. As part of the thesis’ contribution, a prototype of the proposed indoor positioning system is developed using only commodity Wi-Fi hardware and open-source software to evaluate its usability in real-world settings and to demonstrate possible implementation on existing Wi-Fi installations. Experimental results verify the proposed system yields consistent positioning accuracy, even in highly dynamic indoor environments and changing location beacon topologies.

Thesis (MDip (Medical Technology))--Cape Technikon, 1993 / The use of genetic marker testing to resolve cases of disputed paternity, is well established. The number and range of systems used depends on the expertise of the laboratory, and for this reason various laboratories offer different systems. Standard testing includes tests in the following genetic marker systems: human leukocyte antigen (tissue) typing; red cell blood groups; and red cell enzyme and serum protein testing. The Provincial Laboratory for Tissue Immunology currently offers a range of 16 genetic marker systems capable of excluding >99% of falsely accused men. Following the discovery DNA polymorphisms, particularly VNTR DNA polymorphisms, and the commercial availability of VNTR DNA probes, PLTI decided to offer this service to our clients. This study was the initial phase in the establishment of a VNTR DNA typing laboratory and covered the determination of inter-and intra-gel accuracy and precision, selection of restriction enzyme/probe combination, and evaluation and comparison of the results of 100 disputed paternity cases tested using both standard and VNTR DNA typing. Of the 100 cases tested, in 33 cases, the putative father was excluded using standard testing. These exclusions were confirmed using VNTR DNA typing, and, furthermore, an additional two exclusions of paternity were shown using only VNTR DNA typing. In another two cases of disputed paternity, the exclusions obtained using standard tests required further confirmation. VNTR DNA typing convincingly excluded both falsely accused putative fathers. The VNTR DNA typing laboratory now functions as an integral part of the disputed paternity service. Due to the cost and time involved in VNTR DNA typing it is reserved at this stage for: those cases which require further confirmation of the results of standard testing; when the probability of paternity is low (<99.7%); or when a specific request is made.

Made available in DSpace on 2014-06-12T15:49:18Z (GMT). No. of bitstreams: 2 arquivo127_1.pdf: 5135920 bytes, checksum: 7ce3b83de0b3c66828d195aba8d141ea (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2007 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A família Araceae pertence às monocotiledôneas estimando-se que inclua cerca de 3.500 espécies e 105 gêneros, com ampla distribuição mundial, exceto na Antártida. O gênero Anthurium Schott concentra o maior número de espécies (ca. de 800), seguido de Philodendron Schott que inclui cerca de 400 espécies, no qual concentra-se a maioria das análises deste trabalho. A presente avaliação inclui análises citogenéticas de 40 espécies da família Araceae, enquanto 19 espécies foram amostradas pela primeira vez através de marcadores moleculares (DNA Amplification Fingerprinting DAF). Coletas incluíram áreas de floresta amazônica (estados de Manaus e Pará) e de Mata Atlântica incluindo quatro estados brasileiros (Alagoas, Bahia, Minas Gerais e Pernambuco). Para 32 espécies tratam-se das primeiras informações citológicas. O número cromossômico 2n=32 foi observado em 21 espécies, seguido por 2n=34 em oito espécies. O número 2n=30 foi observado apenas em quatro espécies, porém, o mais incomum foi o número 2n=46, observado numa única espécie. Quanto à morfologia cromossômica, observou-se uma conservação entre as populações da maioria das espécies estudadas, com predominância de pares submetacêntricos e metacêntricos, com poucos acrocêntricos. Com base nas atuais análises e em dados da literatura, a disploidia parece ser o principal mecanismo citoevolutivo, sobretudo em Philodendron, o qual pode ser considerado como um gênero paleopoliplóide. Em 13 espécies, a coloração com fluorocromos, antes ou após o tratamento para bandeamento C, evidenciou variações qualitativas (CMA3/DAPI e CB-CMA3 Palavras-chave: Araceae, Cromossomos, Disploidia, Hibridização, Fluorocromos, DNA Amplification Fingerprinting, UPGMA. /DAPI, respectivamente), quantitativas e de posição da heterocromatina constitutiva, revelando tratar-se de um importante marcador carioevolutivo. A impregnação argêntica revelou a variação da quantidade de nucléolos em quatro espécies, que apresentaram no máximo quatro, seis ou oito nucléolos. O presente trabalho apresenta também os primeiro dados com técnicas de bandeamento para Philodendron e Anthurium (e para a família Araceae), assim como hibridização in situ fluorescente (FISH) com sonda de DNAr 45S para A. gracile. Polimorfismos revelados pelo marcador DAF e analisados pelo método de UPGMA permitiram uma distinção entre os 16 taxa analisados, com níveis significativos de polimorfismo. Dendrogramas gerados por esta metodologia foram consistentes com informações taxonômicas e dados populacionais. As relações intra, interespecíficas e intergenéricas (Philodendron e Dieffenbachia) são discutidas, revelando possíveis tendências evolutivas