Thesis (M.S.)--West Virginia University, 2010. / Title from document title page. Document formatted into pages; contains vi, 60 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 58-60).

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 103-112).

Thesis (M. Med. Sc.)--University of Hong Kong, 2005.

Thesis (Ph. D.)--University of Texas at Austin, 2004. / Supervisor: Kerry A. Kinney. Vita. Includes bibliographical references.

Thesis (M. Sc.)--University of Canterbury, 2006. / Typescript (photocopy). Includes bibliographical references (p. 169-200). Also available via the World Wide Web.

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

Molecular genetics use molecular methods that amplify specific fragments of DNA. Today, the molecular techniques which were developed for amplification and detection of specific sequences of nucleic acids helped in a great deal to understand the structure of many diseases. Polymerase chain reaction or PCR is a technique that is used for isolation and amplification of a specific sequence of DNA. PCR is an in vitro method that exploits the in vivo procedure of replication of DNA. DNA polymerase is used to amplify a piece of DNA by in vitro enzymatic replication. As PCR progresses, the DNA thus generated is itself used as a template for replication. This sets in motion a chain reaction in which the DNA template is exponentially amplified. With PCR it is possible to amplify a single or few copies of a piece of DNA across several orders of magnitude, generating millions or more copies of the DNA piece. PCR was designed and presented by Dr Kary Mullis (1983).Today PCR has a great range of implementations related to 1) the cloning and the study of gene’s expression 2) the detection of mutations that are responsible for hereditary diseases 3) Criminology, Toxicology and Forensics. / H Μοριακή Γενετική χρησιμοποιεί μοριακές μεθόδους οι οποίες ενισχύουν συγκεκριμένα τμήματα DNA. Σήμερα, οι μοριακές τεχνικές οι οποίες αναπτύχθηκαν για ενίσχυση και ανίχνευση συγκεκριμένων ακολουθιών νουκλεϊνικών οξέων βοήθησαν σε μεγάλο βαθμό στην κατανόηση της δομής πολλών ασθενειών. Η αλυσιδωτή αντίδραση πολυμεράσης είναι μια τεχνική η οποία χρησιμοποιείται για απομόνωση και ενίσχυση μιας συγκεκριμένης ακολουθίας DNA. Η αλυσιδωτή αντίδραση της πολυμεράσης είναι μια διαδικασία που πραγματοποιείται σε ελεγχόμενες συνθήκες έξω απο ζωντανούς οργανισμούς, η οποία εκμεταλλεύεται την αντιγραφή του DNA που πραγματοποιείται μεσα στον ζωντανό οργανισμό. Όσο η PCR εξελίσσεται δημιουργούνται αντίγραφα DNA χρησιμοποιώντας ως πρότυπο το αρχικό DNA. Αυτό ενεργοποιεί μια διαδικασία αλυσιδωτής αντίδρασης ώπου το DNA αναπαράγεται εκθετικά. Με την PCR μπορείς να δημιουργήσεις εκατομμύρια αντίγραφα DNA από μία συγκεκριμένη ακολουθία. Η PCR σχεδιάστηκε και παρουσιάστηκε απο τον Dr Kary Mullis. Σήμερα η PCR έχει μέγαλο εύρος εφαρμογής σε πεδία σχετικά με τη μελέτη της γενετικής έκφρασης την ανίχνευση μεταλλάξεων οι οποίες είναι υπεύθυνες για κληρονομικές ασθένειες, Εγκληματολογία, Τοξικολογία και Ιατροδικαστική.

Made available in DSpace on 2014-08-13T14:50:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-10-16Bitstream added on 2014-08-13T18:01:30Z : No. of bitstreams: 1 000747021_20151016.pdf: 607081 bytes, checksum: bbdf4b1c6ca753edb9c6aef714441073 (MD5) Bitstreams deleted on 2015-10-16T13:54:03Z: 000747021_20151016.pdf,. Added 1 bitstream(s) on 2015-10-16T13:54:46Z : No. of bitstreams: 1 000747021.pdf: 964954 bytes, checksum: 60f92abc05445fbbd653fbb32d487c4c (MD5) / A família Trypanosomatidae compreende um grande número de protozoários parasitas, incluindo importantes agentes etiológicos de doenças humanas. Entre os causadores de doenças, destacam-se Trypanosoma brucei (responsável pela doença do sono), Trypanosoma cruzi (agente causador da doença de Chagas) e Leishmanias sp (causadoras de leishmanioses). Esses parasitas possuem como mecanismo para formação de seus mRNAs, a reação de trans-splicing, que envolve a excisão de íntrons (regiões intergênicas) e a união dos éxons de dois transcritos independentes, SL RNA (spliced leader) e pré-mRNA aceptor. Em outro trabalho realizado no laboratório, a reação de trans-splicing in vitro foi estabelecida utilizando extratos nucleares (livre de células) de formas epimastigotas de T. cruzi e/ou formas procíclicas de T. brucei e como pré-mRNA aceptor, a sequência parcial de alfa tubulina (contendo a região intrônica, splice site e parte do éxon da alfa tubulina) de T. cruzi marcada radioativamente (dados ainda não publicados). O uso de material radioativo é fator limitante, devido a sua alta periculosidade (extremamente nocivo ao homem e ao meio ambiente) e, também, pelo alto custo relativo aos trâmites burocráticos atuais para sua importação. Portanto, neste trabalho reproduziu-se a reação sem o uso de material radioativo, utilizando-se a transcrição reversa/ reação em cadeia da polimerase (RT-PCR) e gel de agarose corado por brometo de etídeo para a análise dos produtos formados. A metodologia permitirá a avaliação da interferência de substâncias tripanocidas, buscando potenciais alvos terapêuticos nas ribonucleoproteínas de parasitas, preservando as células do hospedeiro. / The Trypanosomatidae family includes a large number of protozoan parasites, with importants etiological agents of human diseases. Among the causes of diseases stand out Trypanosoma brucei (responsible for sleeping sickness), Trypanosoma cruzi (Chagas disease causative agent) and Leishmania sp (causing leishmaniasis). These parasites have a mechanism that form their mRNAs, the trans-splicing reaction, which involves the introns (intergenic regions) excision and two exons union of the independent transcripts, RNA SL (spliced leader) and pre-mRNA acceptor. In another laboratory study, the trans-splicing in vitro reaction was established using nuclear extracts (cell-free) from epimastigotes forms of T. cruzi and / or procyclic forms of T. brucei, and as pre-mRNA aceptor was used the alpha tubulin partial sequence (containing the intronic region, splice site and a radiolabelled part of alpha tubulin exon from T. cruzi (unpublished data). The use of radioactive material is a limiting factor, due to its high hazard (extremely harmful to humans and environment), and also by the high cost relative to the current bureaucratic procedures for importation. Therefore, in this work the reaction was reproduced without the use of radioactive material, using the reverse transcription / polymerase chain reaction (RT-PCR) and agarose gel stained with ethidium bromide for analysis of the formed products. The method allows the evaluation of trypanocidal substances interference searching potential therapeutic targets in the parasite ribonucleoprotein, preserving the host cells.

Made available in DSpace on 2014-10-09T12:51:11Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:56:13Z (GMT). No. of bitstreams: 1 11252.pdf: 6025397 bytes, checksum: 922a7a46ef469bfb01cf33facd40b567 (MD5) / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

Magister Chirurgiae Dentium - MChD / This mini-thesis assessed whether a diode laser with a wavelength of 810 ± 10nm can be utilized as an adjunct to conventional management (i.e. scaling, root planing and polishing) of chronic periodontitis during initial phase therapy. Ethical approval and study registration (Reg no: 14/9/6) was finalized prior to commencement of the study. A split mouth randomised control trial was performed on 25 participants (who presented at the Oral Medicine and Periodontology Department of the University of the Western Cape) diagnosed with active, chronic periodontitis. In order to standardise the split mouth design the quadrants 1 & 4 were assessed together as a set and quadrants 2 & 3 were assessed as a set. A set of these quadrants were randomly assigned to either the test or control quadrants after conventional management was performed in all four quadrants. The base line bacterial colony collection (Micro-IDent®-11, Hain Lifescience GmbH, Nehren, Germany) and the clinical parameters were assessed prior to the commencement of conventional management and were reassessed at the 6 week re-evaluation visit. The set of test quadrants were treated with the diode laser as an adjunct to the preceding conventional management. The control quadrant only received the conventional management. Evaluation of the results demonstrated that the diode laser produced no statistical decrease in the bacterial parameters in the periodontal pockets and resulted in a statistical increase of C. showae (Cs) and T. denticola (Td). The clinical parameters resulted in no statistical difference for any clinical parameter, with the exception of the reduction in BOP that was statistically significant (p< 0,05) with the laser as an adjunct. It is the recommendation that within the limitations of this study, that the utilization of the diode laser (810 ± 10nm) as an adjunct at the initial visit had no statistical effect in the reduction of the bacterial parameters nor resulted in an overall improvement of the clinical parameters.