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1
Liu, Ni. "Detection of trait-associated restriction fragment length polymorphisms in chicken." Thesis, McGill University, 1994. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=55509.
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The gene encoding chicken growth hormone (GH) was isolated from a chicken genomic library. The size of the gene was 4 kb. It was digested with PstI and subcloned into pUC18. Three of the PstI fragments were used for restriction fragment length polymorphisms (RFLPs) analysis at the GH locus in two chicken strains (fat and lean line). Four polymorphic sites were detected using a PstI fragment (PII) as a probe. One polymorphism was located at a SacI restriction site (PS1), and three at MspI sites (PM1, PM2 and PM3). A method based on polymerase chain reaction (PCR) was developed for detecting polymorphisms at PM3 site. A fragment of 823 base pairs which contained the PM3 polymorphic site was amplified. Three genotypes (+/+,$-$/$-$ and +/$-$) were distinguished by examining the MspI digested PCR products in either agarose or polyacrylamide gel.Ten anonymous cDNA clones were also isolated from a chicken liver cDNA library and used for RFLPs analysis. Three of these clones were found to be able to detected RFLPs at MspI sites in chicken strains (strain 7, 8, 9, 8R, S and K) indicating that a high frequency of genes are polymorphic and can be used as markers in mapping experiments. One of the three clones was present on a haploid genetic element. Segregation analysis showed that the inheritance of this haploid gene was determined by the genotype of the female parent.
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Chui, Hon-kit. "Restriction fragment length polymorphism analysis of a hospital outbreak of tuberculosis." Click to view the E-thesis via HKUTO, 2001. http://sunzi.lib.hku.hk/hkuto/record/B31970266.
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Chui, Hon-kit, and 徐漢傑. "Restriction fragment length polymorphism analysis of a hospital outbreak of tuberculosis." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31970266.
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Cicek, Mine II. "Genetic Analysis of Quantitative Trait Loci Associated with Seed Sucrose Content Using Molecular Markers in an Interspecific Glycine Cross." Thesis, Virginia Tech, 1997. http://hdl.handle.net/10919/36506.
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Sucrose content is one of the important seed quality traits in soybean, especially for oriental soyfood production. However, little genetic information is available on this quantitative trait yet.
A previous study was conducted on seed sucrose content of soybean using a population of F2-derived lines from an interspecific cross between an adapted high-sucrose (8.3%) G. max breeding line (V71-370) and a low sucrose (1.6%) G. soja plant introduction (PI407162). Nineteen marker loci, mapping to seven linkage groups (A1, A2, E, F, L1, I, and M), were significantly associated with seed sucrose content after screening 178 polymorphic genetic markers, including RFLPs, SSRs, RAPDs and morphological markers. The replicated field experiments were planted in 1993 and 1995.
The objective of my study was to evaluate QTLs associated with seed sucrose content utilizing an additional 153 F2:3 families from the same cross.
DNA samples from the additional families were analyzed with the nineteen genetic markers associated with sucrose in the previous study. Sucrose data were obtained from seeds harvested from a field experiment conducted in 1995. Single factor analysis of variance results for the sucrose data obtained from the 153 F2:3 families were compared to the 1995 data for the 144 F2:3 families of the previous study.
Of the nineteen genetic markers significantly associated with seed sucrose content in the previous study, seven were also significantly associated in this study. These genetic markers include sgA458a on linkage group A2, NBS61 on linkage group E, sgB164, R-B4a and sgB162 on linkage group L1, and R-B4b and sgA144 on linkage group I. The percent phenotypic variation explained by significant individual markers varied from 2.9 to 6.8% in the 153 F2:3 families.
This study shows that seed sucrose content, a quantitative trait, may be improved using the molecular marker technology. Further research is necessary in different genetic backgrounds of G. max in order to implement these markers in a breeding program for selection.Master of Science
5
Gysling, Kevin. "Polymerase chain reaction restriction fragment length polymorphism identification of trebouxia lichen photosymbionts." Honors in the Major Thesis, University of Central Florida, 2009. http://digital.library.ucf.edu/cdm/ref/collection/ETH/id/1269.
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This item is only available in print in the UCF Libraries. If this is your Honors Thesis, you can help us make it available online for use by researchers around the world by following the instructions on the distribution consent form at http://library.ucf.edu/Systems/DigitalInitiatives/DigitalCollections/InternetDistributionConsentAgreementForm.pdf You may also contact the project coordinator, Kerri Bottorff, at kerri.bottorff@ucf.edu for more information.Bachelors
Medicine
Molecular Biology and Microbiology
6
Gray, Stephen James. "The genotyping of Neisseria meningitidis by restriction fragment length polymorphism (RFLP) analysis." Thesis, Manchester Metropolitan University, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.360085.
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Stevens, Tracy Alison. "Analysis of mitochondrial DNA restriction fragment patterns in killer whales, Orcinus orca." PDXScholar, 1989. https://pdxscholar.library.pdx.edu/open_access_etds/3928.
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The mitochondrial DNA restriction fragment patterns of killer whales (Orcinus orca) were investigated in order to determine the level of genetic differentiation that exists between killer whales from various geographic locations. Twenty one killer whales were examined, seventeen of which were captive killer whales that originated from the North Atlantic and Northeast Pacific Oceans. Two were captive-born animals and two were killer whales that stranded along the Northeast Pacific coast.
DNA was extracted from blood and/or tissue samples, cleaved with a variety of restriction endonucleases and the DNA fragments were separated by horizontal agarose gel electrophoresis. The DNA was then transferred to nylon membranes and the killer whale mitochondrial DNA was visualized by hybridization to the complete mitochondrial DNA genome of Commerson's dolphin (Cephalorhynchus commersonii). The resultant restriction fragment patterns were analyzed to determine whether mitochondrial DNA variation was present between killer whales from different geographic regions or between communities and pods of killer whales from the same geographic location.
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Ogawa, Osamu. "Molecular Genetic Analysis of Human Renal Tumors by Using Restriction Fragment Length Polymorphisms." Kyoto University, 1994. http://hdl.handle.net/2433/160715.
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本文データは平成22年度国立国会図書館の学位論文(博士)のデジタル化実施により作成された画像ファイルを基にpdf変換したものであるKyoto University (京都大学)
0048
新制・課程博士
博士(医学)
甲第5794号
医博第1587号
新制||医||590(附属図書館)
UT51-94-R318
京都大学大学院医学研究科外科系専攻
(主査)教授 野田 亮, 教授 佐々木 正夫, 教授 吉田 修
学位規則第4条第1項該当
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Zhang, Jianhua. "Restriction fragment length polymorphism analysis of chloroplast DNA, mitochondrial DNA, and ribosomal DNA in turfgrasses." Diss., This resource online, 1994. http://scholar.lib.vt.edu/theses/available/etd-06062008-170748/.
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Guo, Haibin. "Combining Conventional Tests and Terminal Restriction Fragment Analysis to Evaluate Microbial Quality of Raw Milk." DigitalCommons@CalPoly, 2011. https://digitalcommons.calpoly.edu/theses/468.
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Abstract
Combining Conventional Tests and Terminal Restriction Fragment Analysis to Evaluate Microbial Quality of Raw Milk
Haibin Guo
The dairy industry is an important part in the domestic economy in the U.S. and the quality of dairy products hinges on raw milk quality. Microorganisms play a critical role in raw milk quality and they are currently tested and monitored by conventional microbiological tests. Some of the most common conventional tests include somatic cell count (SCC), standard plate count (SPC), coliform count (CC), lab pasteurized count (LPC) and proteolytic strain count (PSC). However, these methods do not correlate with each other or with the quality of milk and milk products. One factor that contributes to this lack of correlation is the insufficient knowledge of microbial communities in raw milk. In this work we aimed to evaluate modern molecular methods to complement traditional quality procedures that may eventually complement conventional tests and improve milk quality evaluation. Therefore, a molecular method, Terminal Restriction Fragment (TRF) analysis was introduced. TRF analysis has been widely used as a tool to investigate the microbial communities in environmental samples. In this study, it was applied to investigate the microbial communities in raw milk and evaluate raw milk quality.
Milk samples were collected for over six months in the Cal Poly dairy farm and evaluated by conventional tests and TRF analysis. Samples were defined as “high quality” milk and “low quality” milk according to each conventional test first. The cutoffs applied were: 50,000 cfu/ml for SPC, 70,000 cells/ml for SCC, 100 cfu/ml for CC and 250 cfu/ml for LPC. TRF analysis was conducted on raw milk samples subsequently. DNA extraction was optimized. Non-Parametric Multivariate Analysis of Variance (NPMANOVA) was applied to TRF profiles from low and high quality milk. The analysis of Similarity of Percentage (SIMPER) was used to determine each TRF peak’s contribution to the dissimilarity between the profiles of high and low quality milk. The genus/species represented by TRF peaks were estimated via database matching. In addition, conventional tests and TRF analysis were also used to analyze the factors causing low quality milk. Rain event and cow’s apparent health were the two factors investigated since raw milk samples were collected from cows in different apparent health status on wet days and dry days.
Conventional tests revealed strong correlations between the results of SPC and PSC, and SPC and CC (coefficients of correlation > 0.8). It implied that the results of conventional tests might not be independent, so the statistics based on the assumption of independence of variables were not suitable to analyze the data. SCC showed no strong correlation with any other conventional tests. Raw milk samples were grouped as high quality and low quality according to SPC, CC, SCC and LPC. Using TRF analysis, it was found that there was a significant difference between TRF profiles from low and high quality milk when the quality was defined by SPC or LPC. A TRF peak at 268 bp generated by DpnII was predominant in the TRF profiles and had high abundance in the profiles of low quality milk. Hence, Pseudomonas spp. represented by TRF peak at 268 bp was likely the predominant bacteria in the microbial community associated with raw milk. TRF peaks at 61 bp, 81 bp, 104 bp, 104 bp, 201 bp, 242 bp, 268 bp, and 270 bp contributed the most to the dissimilarity between TRF profiles from different groups of samples. In addition, the presence of DNA derived from viable but non-culturable species that were associated with raw milk quality was detected.
Rain event was the most important factor affecting the microbial quality of raw milk in this study. Both the conventional tests and TRF analysis showed that there was a significant difference between raw milk samples collected on wet days versus dry days. Samples collected on wet days harbored high bacterial counts and had high abundance of the predominant TRF peaks. In addition, the same TRF peaks contributing the most to the dissimilarity between groups separated by rain event were found to be among those contributing the most to the dissimilarity between groups of high and low quality milk defined by conventional tests. During wet days, the low quality milk was likely caused by the increased dirtiness of cow’s teats. Soil microbes are often associated with microorganisms in raw milk such as psychrotrophic bacteria, coliform groups and spore-formers. Cow’s apparent health status showed no significant influence on the microbial quality of raw milk.
Overall, the combination of conventional tests and TRF analysis can yield a comprehensive understanding of microbial community in raw milk and improve the evaluation of raw milk quality. TRF analysis was demonstrated as a useful tool and a complement to conventional tests for milk quality evaluation by providing more information on the microbial community associated with raw milk. Findings in this study can offer a basis for further study and may help the dairy industry improve raw milk quality evaluation system.
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VINCENT, ILLANDI PASCALE. "Mise en place d'une etude de liaison : a propos de la maladie de rendu osler." Lyon 1, 1992. http://www.theses.fr/1992LYO1M021.
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Zhu, Jiangtao. "Genetic Analysis of Experimental and Commercial Turkey Lines Using Restriction Fragment Length Polymorphism and DNA Fingerprinting /." The Ohio State University, 1995. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487931993466504.
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Ribeiro, Camila Maria Beder 1982. "Caracterização clínica-histológica e estudo de polimorfismos das respostas T-Helper-1 e 2 (Th1/2) nas doenças imunologicamente mediadas com manifestações bucais = líquen plano oral e reação liquenóide oral por amálgama dental." [s.n.], 2012. http://repositorio.unicamp.br/jspui/handle/REPOSIP/289239.
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Orientador: Jacks Jorge JuniorTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: As lesões liquenóides orais (LLO), como líquen plano oral (LPO) e reação liquenóide oral por amálgama dental (RLO-AD), são doenças imunologicamente mediadas (DIM) com características histopatológicas semelhantes, porém com prognósticos distintos. As suas etiopatogenias têm sido relacionadas a polimorfismos gênicos e secreção citocinas de inflamatórias (CI), as quais são responsáveis pelas respostas imunológicas inadequadas, que causam danos estruturais à mucosa oral (MO) e estimulam o recrutamento de células inflamatórias. Assim, foi realizado um estudo em voluntários portadores de LLO (grupo-1, n=39), em voluntários saudáveis (grupo-2, n=39), e em cem amostras de DNA provenientes de voluntários saudáveis (grupo-3, n=100). A pesquisa teve como objetivos selecionar e diferenciar casos de LLO clássicas (LLO-c) por meio dos critérios de diagnósticos de LLO da Organização Mundial de Saúde (Cdx-LLO-OMS); determinar a hipossalivação nesses pacientes; analisar outros achados histopatológicos presentes nas LLO; quantificar linfócitos T (LT-CD3, LT-CD8), linfócitos B (LB), plasmócitos, mastócitos, imunomarcados contidos no infiltrado inflamatório subepitelial (IISE) nos casos de LLO a fim de avaliar as respostas Th1/2; e estimar a frequência dos polimorfismos dos genes Th1/2 envolvidos na síntese de CI através da reação em cadeia da polimerase (PCR) e polimorfismo no comprimento do fragmento de restrição (RFLP). De forma geral, pacientes do gênero feminino com idade média de 53,13 anos apresentaram maior frequência de LLO (p=0,06; OR:2,57). O infiltrado inflamatório perivascular profundo e a presença de folículos linfóides foram frequentes nas RLO-AD (p<0,001). A contagem de LB foi maior nas RLO-AD (p<0,05). A hipossalivação foi frequente em portadores de LLO (p<0,001;OR:19,72). Os alelos mutados IL4-590T,TNF-?-308A, e IL10592C foram freqüentes nos portadores de LLO (p<0,0001). Portadores de LPO apresentaram mais alelos mutados IL4-590T e IL10-592C (p<0,05). Concluiu-se, portanto que pacientes do gênero feminino com média de idade de 53 anos apresentam mais LLO. A hipossalivação foi associada à presença da lesão oral em portadores de LLO. As LLO estão associadas aos altos índices de células de reação inflamatória crônica e à expressão gênica das citocinas relacionadas às respostas Th1/2
Abstract: Oral lichenoid lesions (OLL), such as oral lichen planus (OLP) and amalgam-associate oral lichenoid reaction (AAOLR) are immunologically mediated diseases (IMD) with similar histopathologic features but distinct prognoses. Their etiopathogenesis have been related to gene polymorphisms and inflammatory cytokine (IC) secretion, which are responsible for the inappropriate immune responses that cause structural damage to the oral mucosa (OM) and stimulate the recruitment of inflammatory cells. Therefore, this study was conducted in a group of volunteers with OLL (group-1, n=39), another group comprised of healthy volunteers (group-2, n=39), and a third group comprised of hundred DNA samples obtained from healthy volunteers (group-3, n = 100). The research aimed to select and differentiate classic cases of OLL by means of diagnostic criteria for OLL of the World Health Organization (WHO-OLL-CDx), to determine the hyposalivation in these patients, to analyze other histological findings features present in OLL; to evaluate the Th1/2 by means of quantification of inflammatory cells, T-lymphocytes (CD3-TL, CD8-TL), B lymphocytes (BL), plasma cells and mast cells immunostained contained in the subepithelial inflammatory infiltrate (SEII) on the slides of OLL, and estimate the frequency of polymorphisms of genes Th1/2 involved in the synthesis of IC by means of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Overall, female patients with mean age of 53.13 years had a higher frequency of OLL (p = 0.06, OR: 2.57). The deep perivascular inflammatory infiltrate and lymphoid follicles were common in the AAOLR (p <0.001). The LB count was higher in AAOLR (p <0.05). The hyposalivation was common in patients with OLL (p <0.001, OR: 19.72). Mutated alleles of IL4-590T, TNF-?-308A and IL10-592C were frequent in patients with OLL (p <0.0001). Patients with OLP had more mutated alleles IL4 and IL10-590T-592C, when compared to AAOLR (p <0.05). Therefore it was concluded that female patients with a mean age of 53 have more OLL. Hyposalivation was associated with the presence of oral lesions in patients with OLL. The oral lichenoid lesions are associated with high rates of chronic inflammatory cell reaction and gene expression of cytokines related to Th1/2 response
Doutorado
Patologia
Doutor em Estomatopatologia
14
Bijelovic, Jelena. "Identification of mould and blue stain fungi on wood using Polymerase Chain Reaction and Terminal Restriction Fragment Length Polymorphism." Thesis, Uppsala University, Department of Medical Biochemistry and Microbiology, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7100.
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Wood inhabiting fungi oposes a great problem for preservation of wooden surfaces everywhere, being the main problem of economic losses of wooden products.
A reference collection consisting of 9 different genus constituting of 21 different strains of wood-inhabiting fungi was used for identification of unknown species of mould and blue stain fungi on wood. The fungus DNA from the samples was isolated from malt extract agar. PCR (Polymerase Chain Reaction) was conducted on rDNA ITS1 and ITS2 regions for amplification of the DNA. The 21 samples were collected to a reference collection for identification of unknown species of fungi on wooden field samples using PCR and T-RFLP (Terminal Restriction Fragment Length Polymorphism).
PCR-based methods, sequencing and T-RFLP were proven to be simple and
accurate methods for detection and identification of fungi in their early stage.
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Riocreux, Ingrid. "La négation dans le fragment moraliste (La Rochefoucauld, Pascal, Vauvenargues, Chamfort)." Thesis, Paris 4, 2013. http://www.theses.fr/2013PA040154.
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Fait de langue omniprésent et multiforme (impliquant des données morpho-syntaxiques et lexicales), la négation constitue véritablement un stylème de la moralistique classique. Focalisation, incidence, forclusion, négation uniceptive, implicite, préfixale, lexématique, problèmes de polarité et de scalarité : le statut central de la négation permet de saisir un positionnement énonciatif commun aux quatre auteurs du corpus autant qu’une prise de position philosophique spécifique à chacun d’eux. L’ambiguïté pragmatique communément associée à la négation est levée, dès lors que l’on admet la possibilité d’une lecture à plusieurs niveaux : la négation descriptive correspond à la représentation traditionnelle du discours moraliste comme une parole solitaire, isolée entre deux blancs typographiques. Mais précisément, lorsqu’on considère cette parole comme un discours adressé, la dimension polémique de la négation apparaît pleinement, plus précisément sa portée contre-doxique et métalinguistique. Les moralistes dénoncent le caractère faussé du discours commun ; ils conçoivent la négation comme un moyen de rompre le lien de référentialité abusif établi par celui-ci entre des concepts moraux et des comportements qui n’ont de vertueux que l’apparence. De l’héritage apophatique, sensible chez Pascal, jusqu’au prénihilisme chamfortien, en passant par l’anthropologie négative de La Rochefoucauld et l’immanentisme anti-artificialiste de la morale chez Vauvenargues, la négation offre une grille de lecture nouvelle pour étudier l’évolution du genre moralisteBased on quantitative data, this study shows how essential negation is in the understanding of moralistique as a literary genre that can be identified as such through precise formal elements. I examine many aspects of negation, including the questions of scope, internal and external negations, restricted negation, forclusion, implicit negation, prefixal and lexematic negation and polarity scales. Not only is negation a linguistic scheme (involving various morpho-syntactic as well as lexical patterns) but it also works as a stylistic device which the moralists make a constant and specific use of. Whereas it is commonly held that negation is pragmatically ambiguous, I argue that, in focusing on the moralist as a spectator of society, the critiques have implicitly considered negation to be mostly descriptive. While correct, this interpretation should be qualified. The main aspect of the moralists’ negation rests in its polemical power. The moralists intend to rectify a biased use of words resulting from a false conception of moral values. Therefore, these writers do not say what things are as much as what they are not. From Pascal’s apophatic views, through La Rochefoucauld’s negative anthropology and Vauvenargues’ refusal of artificial morality, to Chamfort’s prenihilistic philosophy, negation appears as a new way to get a better understanding of the evolution of moralistique
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Wallace, Diane Marie. "Large restriction fragment pattern analysis of genomic Mycobacterium fortuitum DNA as an epidemiological tool in the investigation of institutional outbreaks." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp04/mq22417.pdf.
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Masli, Aryananda. "Search for restriction fragment length polymorphism of Phaseolus vulgaris in relation to the immune gene to bean common mosaic virus." Thesis, University of North Texas, 1991. https://digital.library.unt.edu/ark:/67531/metadc798405/.
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A technique involving Restriction Fragment Length Polymorphism (RFLP) was used to observe the DNA fragment polymorphism between a bean cultivar with I/I genotype and a bean cultivar with i/i genotype. The I gene encodes immunity to bean common mosaic virus (BCMV).
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Pavan, Tycha Bianca Sabaini 1983. "Detecção do gene 16Sr-RNA e identificação de patógenos bacterianos, causadores de sepse neonatal precoce, pela técnica da RFLP-PCR ("Restriction Fragment Length Polymorfism - Polymerase Chain Reaction")." [s.n.], 2013. http://repositorio.unicamp.br/jspui/handle/REPOSIP/309724.
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Orientador: Sergio Tadeu Martins MarbaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas
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Resumo: O objetivo do trabalho foi descrever e avaliar a capacidade da "RFLP-PCR" para detectar patógenos bacterianos em recém-nascidos (RN) e determinar seu uso diagnóstico na sepse neonatal precoce (SNP). Foram avaliados RNs assintomáticos, filhos de gestantes com fatores de risco para infecção ovular e RNs com apresentação de sintomas clínicos sugestivos para SNP, nas primeiras 48 horas de vida, atendidos na unidade neonatal do CAISM/UNICAMP/SP. Realizou-se extração genética das amostras de 200?L de sangue total através do kit QIAamp DNA Mini kit column (Qiagen), seguida da detecção do gene universal bacteriano 16 S rRNA e identificação microbiana pelas sucessivas digestões com enzimas de restrições - HaeIII, AluI, DdeI, MnlI. Foram avaliados 65 RN assintomáticos e 178 RN com apresentação de sinais clínicos sugestivo de SNP. Os patógenos detectados foram K. pneumoniae, S. pneumoniae, L. monocytogenes, P. aeruginosa, E.coli, S.pyognes, S. agalactiae, S. epidermidis e S. aureus. Resultados duvidosos ou bactérias não identificadas ocorram em 56 amostras. A positividade da "RFLP-PCR" foi de 55,3% no grupo assintomático e 51,6% no grupo sintomático, sem diferença estatística (p=0,583). Houve comprovação de sete RN com sepse por cultura e 24 com sepse clínica e foram encontrados valores de sensibilidade de 87,8%, especificidade de 46,6%, valores preditivo positivo de 23,3% e preditivo negativo de 95,4%. Em conclusão, o uso da técnica mostrou uma taxa de acurácia baixa para o diagnóstico para a SNP satisfatória
Abstract: Study objective was to describe and evaluate the ability of RFLP-PCR for detection of bacterial pathogens in newborn and to determine its diagnostic utility in early neonatal sepsis. There were evaluated asymptomatic newborns, children of mothers with risk factors for infection ovulate and newborns with presentation of clinical symptoms suggestive of early neonatal sepsis, occurring before 48 hours of life, evaluated at neonatal unit at CAISM-UNICAMP. Whole blood samples were taken from newborns - 200 ?L each. A genetic extraction were performed using QIAamp DNA Mini kit column (Qiagen) and also the detection of bacterial universal 16 S rRNA gene, subsequent microbial identification using restriction digestion enzymes - HaeIII, AluI, DdeI, MnlI. There were 65 asymptomatic infants and 178 symptomatic newborns. The identified bacteria were K. pneumoniae, S. pneumoniae, L. monocytogenes, P. aeruginosa, E.coli, S.pyognes, S. agalactiae, S. epidermidis and S. aureus. Uncertain or unidentified pathogens occurred in 56 samples. RFLP-PCR positivity was 55,3% in the asymptomatic group and 51,6% in the symptomatic group, with no statistical difference (p=0,583). There were culture comproved sepsis in 7 infants and in 24 newborns were made a diagnostic of clinical sepsis. Diagnostic indexes were: sensibility 87.8%, specificity 46.6%, positive predictive value 23.3% e negative predictive value 95.4%. In conclusion, RFLP-PCR had an unsatisfactory accuracy index early neonatal sepsis
Mestrado
Saude da Criança e do Adolescente
Mestra em Ciências
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Tonin, Mariana Ferreira 1978. "Caracterização taxonômica de espécies do gênero Xanthomonas." [s.n.], 2012. http://repositorio.unicamp.br/jspui/handle/REPOSIP/317296.
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Orientador: Suzete Aparecida Lanza DestéfanoTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: Espécies pertencentes ao gênero Xanthomonas são responsáveis por doenças que podem causar grandes perdas econômicas em diversas culturas. Esses fitopatógenos têm sido objeto de diversos estudos taxonômicos resultando em significativas alterações na classificação em nível inter e infraespecífico. O presente estudo teve por objetivo caracterizar taxonomicamente bactérias do gênero envolvendo: (1) diferenciação e análises filogenéticas de espécies do gênero Xanthomonas; (2) esclarecimento da posição taxonômica de linhagens classificadas como Xanthomonas sp.; (3) diferenciação de patovares da espécie X. campestris; (4) desenvolvimento de primers específicos para X. campestris, X. translucens, X. cucurbitae e X. melonis. O par de primers rpoB2F/rpoB3R, desenhado a partir de sequências do gene rpoB, foi empregado em experimentos de amplificas;ao utilizando-se DNAs de 26 espécies do gênero Xanthomonas e os produtos de amplificas;ao (800 pb) foram digeridos com diversas endonucleases. Os perfis de restrição obtidos com a utilização da enzima Hae III permitiram a diferenciação da maioria das espécies, incluindo os patógenos de mesmo hospedeiro como X. albilineans e X. sacchari (patogênicas à cana-de-açúcar); X. cucurbitae . e X. melonis (patogênicas ao melão); X. vesicatoria, X. gardneri e X. euvesicatoria/X. perforans (patogênicas ao tomateiro). Ainda, os produtos de amplificação do gene rpoB foram seqüenciados e a árvore filogenética construída a partir destas sequências também possibilitou a diferenciação das espécies do gênero, indicando que o gene rpoB pode ser considerado um marcador molecular eficiente para o estudo das relações filogenéticas de espécies do gênero Xanthomonas. Os DNAs de linhagens classificadas como Xanthomonas sp. também foram analisados por meio de experimentos de amplificação com primers específicos para a espécie X. axonopodis, de hibridizas;ao DNA-DNA, e analise de multilocus utilizando-se sequências dos genes rpoB, rpoA, atpA, recA e região espaçadora 16S-23S DNAr. Os resultados mostraram que as linhagens de X. sp. pv. viticola, X. sp. pv. betae e X. sp. pv. paulliniae pertencem a espécie X. axonopodis, entretanto, não foi possível definir a posição taxonômica das linhagens de X. sp. pv. arracaciae, X. sp. pv. esculenti e X. sp. pv. eucalypti, embora várias ferramentas tenham sido utilizadas. Assim, somente estudos complementares deverão ser conduzidos visando esclarecer a classificação dessas linhagens. Nesse estudo também foram analisadas as espécies X. campestris, X. translucens, X. cucurbitae e X. melonis. Visando a diferenciação dos patovares da espécie X. campestris, os DNAs destas linhagens foram submetidos a experimentos de PCR-RFLP do gene rpoB e as digestões duplas utilizando-se Cfo I/Mbo I. Os resultados permitiram a diferenciação dos patovares X.c. pv. raphani, X.c. pv. barbariae, X.c. pv. incanae, X.c. pv. armoraciae e X.c. pv. campestris/ X.c. pv. aberrans. Ainda, os dados obtidos nas amilises do gene rpoB permitiram o desenvolvimento de primers específicos para as espécies X. campestris, patogênicas as crucíferas (rpoB2F/xcamR) e X. translucens, patogênicas a gramíneas e cereais (trans1F/trans2R). Alem disso, amilises de sequências da região espaçadora 16S-23S DNAr possibilitaram o desenho do par de primers mecF/mecR, especifico para X. cucurbitae e X. melonis, espécies patogênicas ao melão. A especificidade destes primers foi confirmada em experimentos de amplificação utilizando-se DNAs de algumas bactérias de diferentes gêneros isoladas de mesma espécie de plantas hospedeiras. O nível de sensibilidade da técnica de PCR utilizando-se os primers desenvolvidos foi de 0,1 pg para rpoB/xcam, de 0,01 ng para trans1F/trans2R e de 1 pg para mecF/mecR
Abstract: Xanthomonas species are responsible for diseases causing economic losses in many crops. These phytopathogens have been subject of several taxonomic studies resulting in significant changes at interespecific or infraspecific level. This study aimed the taxonomic characterization of Xanthomonas species including: (1) differentiation and phylogenetic analysis of Xanthomonas species; (2) clarification of taxonomic position of strains classified as Xanthomonas sp.; (3) differentiation of the pathovars of X. campestris; ( 4) development of specific primers for X. campestris, X. translucens, X. cucurbitae and X. melonis. The rpoB2F/rpoB3R primers, designed from rpoB gene sequences, were employed in amplification experiments using DNAs from 26 species of Xanthomonas genus and the products (800 bp) were digested with different restriction enzymes. Profiles using Hae III allowed to differentiate the most species of the genus, including pathogens the affect the same plant host as X. albilineans e X. sacchari (pathogenic to sugarcane); X. cucurbitae e X. melonis (pathogenic to melon); X. vesicatoria, X. gardneri and X. euvesicatoria/X. perforans (pathogenic to tomato). Amplification products of the rpoB gene were sequenced and the phylpgenetic tree constructed from these sequences also allowed the differentiation of the Xanthomonas species, indicating that the rpoB gene can be used as an efficient molecular marker for phylogenetic relationships studies within the Xanthomonas genus. DNA of the strains classified as Xanthomonas sp. were also analysed by the amplification experiments using specific primers for X. axonopodis species, DNA-DNA hybridization and multilocus sequence analysis of the genes rpoB, rpoA, atpA, recA and intergenic spacer region 16S-23S rDNA. Results showed that the strains of X. sp. pv. viticola, X. sp. pv. betae and X. sp. pv. paulliniae belong to X. axonopodis species, however, it was not possible to define the taxonomic position of X. sp. pv. arracaciae, X. sp. pv. esculenti and X. sp. pv. eucalypti, although different approaches have been used. Thus, further studies should be conducted in order to clarify their classification. In this study X. campestris, X. translucens, X. cucurbitae and X. melonis were also analyzed. In order to differentiate the pathovars of the X. campestris specie, the DNAs of these strains were submitted to PCR-RFLP analysis of the rpoB gene and the double digestions using Cfo I/Mbo I allowed the differentiation of the pathovars X. c. pv. raphani, X.c. pv. barbariae, X.c. pv. incanae, X.c. pv. armoraciae and X.c. pv. campestris/ X.c. pv. aberrans. Also, the data obtained in the rpoB gene analysis allowed the development of specific primers for the species X. campestris, pathogenic to crucifers (rpoB2F/xcamR) and X. translucens, pathogenic to grasses and cereals (trans1F/trans2R). Furthermore, intergenic spacer 16S-23S rDNA sequences analysis enabled the development of the mecF/mecR primers, specific for X cucurbitae and X melonis, both pathogenic to melon. The specificity of these primers was confirmed by amplification experiments using DNAs from bacteria belonging to different genera pathogenic to the same plant hosts. Sensitivity level of the PCR technique using the primers developed was 0,1 pg for rpoB/xcam, 0,01 ng for trans1F/trans2R and 1 pg for mecF/mecR
Doutorado
Genetica de Microorganismos
Doutor em Genetica e Biologia Molecular
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Luro, François. "Utilisation des marqueurs moléculaires pour la cartographie du génome et les études génétiques chez les agrumes." Bordeaux 2, 1993. http://www.theses.fr/1993BOR28270.
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Vasilache, Gouandjika Ionela. "Circulation et dérive génétique du poliovirus en Afrique Centrale et de l'ouest." Paris 6, 2006. http://www.theses.fr/2006PA066222.
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Smith, Robert C. "Ecological Factors in Design of a Two-Sludge Nitrifying Activated Sludge System Incorporating Side-Stream Treatment of Anaerobic Digester Supernatant." University of Cincinnati / OhioLINK, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1291307830.
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Starr, Terence. "Isolation and characterization of DNA probes from the short arm of the human X chromosome which detect restriction fragment length polymorphisms." Thesis, University of British Columbia, 1986. http://hdl.handle.net/2429/26082.
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The isolation and characterization of DNA probes which detect RFLPs from a small region of the human genome, Xp, has been the focus of this thesis. Thirty X specific DNA probes were isolated from a flow sorted library constructed from a 49,XXXXY cell line. These probes were confirmed to be from the X chromosome by hybridization to dot blots containing DNA from mouse, human, and a somatic cell hybrid which contained the X chromosome as its only human DNA. Hybridization to somatic cell hybrids containing different regions of the X chromosome localized 9 of the probes to Xp. After testing 185 probe-enzyme combinations three of the Xp probes (localized to the distal 2/3 of Xp) were found to detect RFLPs. One probe, pTS316, was shown not to be inherited in a Mendelian fashion. The presence of one of the polymorphic bands detected by this probe correlated with the phenomenon of X chromosome inactivation.Medicine, Faculty of
Medical Genetics, Department of
Graduate
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Vadari, Yoganand. "Estimation of Microbial Diversity in Poultry Litter Using Terminal Restriction Fragment Length Polymorphism and Isolation of Phosphate Accumulating Bacteria from Poultry Litter." TopSCHOLAR®, 2004. http://digitalcommons.wku.edu/theses/239.
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The contamination of fresh water by phosphates in poultry litter results in substantial eutrophication of fresh water causing fish kills and other types of environmental damage. The poultry indus try in Kentucky is expanding rapidly. The number of broilers is increasing as more poultry farms are established in the state producing waste that needs disposal. Investigations were made to study the possibility of using microorganisms normally found in poultry litter to sequester phosphate, thereby delaying phosphate runoff after litter is applied to croplands. Little is known, however, about the microflora of poultry litter. Terminal restriction fragment length polymorphism of 16S rDNA from bacteria was used to investigate the bacterial diversity of poultry litter. Poultry litter was collected from a local producer. DNA was isolated using commercial kits and amplified using the polymerase chain reaction with primers specific for bacterial 16S rDNA. The amplified fragments were digested using HhaI restriction endonuclease and the DNA fragment lengths were determined. To determine the sensitivity of this method, known quantities of Escherichia coli cells were spiked into litter prior to DNA extraction. Successful amplification of the bacterial rDNA was highly variable but could be improved by passing the purified DNA through two purification columns in lieu of only one column. The detection threshold for E. coli was 10 cells, however, the results also varied widely. Bacteria capable of hyper-accumulating intracellular phosphate were isolated from poultry litter as possible tools for phosphate remediation in poultry litter. Five strains of phosphate accumulating bacteria were successfully isolated from poultry litter. Poultry litter was suspended in sterile nanopure water and 100μl was plated on BHI plates containing an addtional 750mM K2HPO4. Isolated colonies were screened for intracellular metachromatic granules using the Nile blue stain, a presumptive test for polyphosphate. Positive colonies were cultured in BHI and BHI with supplementation of K2HPO4 and free intracellular phosphate concentrations were determined in cell extracts. Total phosphates were measured in cell extracts subjected to hydrolysis by addition of 12N HCl and heating at 100°C for 60 min. Polyphosphate was determined by subtraction of free phosphates from total phosphates. Results showed five isolates of gram-positive bacteria were obtained from poultry litter. All isolates were cocci arranged in chains or clusters and were catalase positive. All isolates showed considerable levels of intracellular phosphate accumulation, which were comparable to Microlunatus phosphovorus, a bacterium known to hyper-accumulate phosphate. Biolo g analysis indicated four of the five strains isolated were Staphylococcus sp. and one strain was unidentified.
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Ishii, Takashige. "CYTOPLASMIC AND NUCLEAR GENOME DIFFERENTIATION IN A-GENOME DIPLOID SPECIES OF RICE AS REVEALED BY THE RESTRICTION FRAGMENT LENGTH POLYMORPHISM ANALYSIS OF DNAS." Kyoto University, 1991. http://hdl.handle.net/2433/78244.
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Kirker, Grant Terral. "EFFECTS OF CHLOROTHALONIL AND BUTYLATED HYDROXYTOLUENE ON MICROBIAL COMMUNITIES INVOLVED IN THE DETERIORATION OF WOOD USING TERMINAL RESTRICTION FRAGMENT LENGTH POLYMORPHISM (T-RFLP) ANALYSES." MSSTATE, 2008. http://sun.library.msstate.edu/ETD-db/theses/available/etd-04022008-155301/.
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The effects of an organic biocide (CTN) with and without co-added antioxidant (BHT) on microbial communities in SYP were assessed using terminal restriction fragment length polymorphism (T-RFLP) analyses in both field and accelerated decay laboratory studies. Ammoniacal copper quaternary (ACQ-C) was used as a positive control in the field study component, but not in the laboratory test. Field stakes were treated with 0.25 and 0.37% ammoniacal copper quat (ACQ-C), CTN (0.1 and 0.25%), CTN (0.1 and 0.25%) with 2% BHT added, 2% BHT alone, and controls were left untreated. In the field studies, preservative treatment slowed the initial colonization of wood by fungi. Higher species richness and diversity were found in non-biocidal treatments (BHT and untreated controls). Fungal communities in treated wood were different based on their species composition, but eventually became more similar to untreated controls. Preservative treatment increased richness and diversity of basidiomycete fungi, but overall presence of basidiomycetes was low compared to other fungi. Preservatives did not change the species composition of basidiomycetes compared to untreated controls. Preservative treatment initially increased bacterial richness and diversity, but over time these trends diminished to levels consistent with untreated controls. Preservatives changed the species composition of colonizing bacteria so that treated and untreated communities remained different over 15 months of soil exposure. Bacterial diversity was negatively correlated with CTN depletion at the lowest rate. In the accelerated decay laboratory test, the effects of CTN and/or BHT on bacterial, fungal, and basidiomycete communities in composted and uncomposted soil were evaluated over a 12 month period. Composted soil had less fluctuation in changing microbial diversity due to more constant moisture. The consensus of the analyses of the bacterial, fungal, and basidiomycete communities indicate that wood preservatives increased microbial species richness and diversity. Preservative treatment increased species turnover that decreased over time. Eventually, microbial communities approached a stable community structure consistent with untreated controls. Preservatives were completely degraded after 30 days exposure; however, definite changes in bacterial and fungal richness, diversity, and species composition were found. Basidiomycetes again represented the smallest portion of the microbial community involved in the overall decay process.
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Glavan, Tiffany Wallingford. "Molecular Analysis Reveals Unique Microbiome in Ileal Pouch During Pouchitis Compared to Healthy Pouches in Ulcerative Colitis and Familial Adenomatous Polyposis." DigitalCommons@CalPoly, 2011. https://digitalcommons.calpoly.edu/theses/543.
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In severe cases of ulcerative colitis (UC) unresponsive to current treatment options, patients require a complete proctocolectomy, or surgical removal of the colon. Ileal pouch anal anastomosis (IPAA) has become the preferred surgical technique for patients who require surgery, as this method restores rectal function. This procedure is also used to treat colorectal cancers such as adenocarcinoma and familial adenomatous polyposis (FAP). The surgery involves an abdominal colectomy with the construction of an ileal pouch created from folded tissue recovered from the ileal portion of the small intestine. Up to 50% of patients who require IPAA surgery experience an episode of pouchitis, a non-specific inflammation of the constructed ileal pouch with unknown etiology. Several hypotheses have been proposed regarding the pathogenesis of pouchitis. Current theories include bacterial overgrowth due to fecal stasis, microbial imbalance (dysbiosis), immune alteration, genetic susceptibility, metaplasia, ischemic complications of surgery, a recurrence of UC, or even a novel form of inflammatory bowel disease. The efficacy of antibiotics and probiotics in treating pouchitis and maintaining remission underscores the importance of gut microbiota in the development of this condition. In the study, we aimed to characterize the intestinal bacterial communities that inhabit IPAA pouches of both UC and FAP patients, in an effort to investigate the hypothesis that bacterial dysbiosis is involved in the pathogenesis of pouchitis. Mucosal biopsy and stool samples were analyzed from patients with UC and pouchitis (UCP), healthy UC controls (HUC) and healthy pouches with a background of FAP (FAP). Samples were examined through analysis of terminal restriction fragment length polymorphisms (TRF) and DNA sequencing. The data presented here demonstrate that a microbial imbalance exists in pouchitis, as bacterial communities in pouchitis differ significantly from healthy UC pouches and pouches constructed for FAP. Both methods identified potential groups of organisms that may play a role in the development of pouchitis, including decreases in protective Lactobacillus and Bacteroides and increases in mucin-degrading Clostridium and Akkermansia. A better understanding of the factors driving the pathogenesis of pouchitis will not only benefit patients with this disease, but also lead to a better understanding of the complex relationship that exists between the human host and the diverse community of organisms that inhabit the gastrointestinal tract.
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Saito, Daniel 1974. "Caracterização das comunidades bacterianas associadas as infecções endodonticas : abordagem independente de cultivo." [s.n.], 2007. http://repositorio.unicamp.br/jspui/handle/REPOSIP/288628.
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Orientador: Reginaldo Bruno GonçalvesTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: A presente tese teve como objetivo a caracterização das comunidades bacterianas associadas às infecções endodônticas pelo emprego de técnicas moleculares independentes de cultivo. Ao todo, foram analisadas amostras intra-radiculares provenientes de 34 elementos dentários associados a infecções endodônticas. A análise de bibliotecas clonais de DNA ribossomal 16S (16S rDNA) permitiu a identificação de 2 a 14 filotipos bacterianos (espécies) por elemento dentário (média= 9,6), perfazendo um total de 46 filotipos distintos. Dentre estes, 9% foram considerados previamente desconhecidos e classificados taxonomicamente como novos membros da ordem Clostridiales. Espécies reconhecidamente endodônticas dos gêneros Bacteroides, Campylobacter, Eubacterium, Peptostreptococcus, Selenomonas, Treponema e Veillonella foram detectadas, assim como representantes de gêneros menos freqüentemente descritos, como Burkholderia, Filifactor e Megasphaera. O emprego da técnica quantitativa de PCR em Tempo Real, possibilitou a detecção de P. gingivalis, T. forsythia e a coexistência de ambas em 24%, 56% e 18% dos pacientes avaliados, respectivamente. Nenhuma correlação significativa foi evidenciada entre os níveis de ambas as espécies, individualmente ou em conjunto, e a presença de sintomatologia dolorosa. O uso de T-RFLP na avaliação da estrutura das comunidades bacterianas revelou um total de 123 (endonuclease HhaI) e 122 (endonuclease MspI) fragmentos de restrição terminais (T-RFs) distintos, com médias de 20,8 e 20,0 T-RFs por elemento dentário, respectivamente. Aproximadamente 50% dos fragmentos detectados apresentaram-se, no máximo, em 2 pacientes, indicando uma alta variabilidade na composição microbiana. As análises de clusterização e de estatística multivariada não revelaram diferenças significativas nas comunidades bacterianas entre os grupos de estudo assintomático, sensível ao toque e sintomático. De modo geral, os resultados obtidos reiteraram o conceito de que a microbiota associada às infecções endodônticas é essencialmente polimicrobiana, altamente variável entre indivíduos, e constituída predominantemente por bactérias anaeróbias Gram-positivas do filo Firmicutes. As espécies P. gingivalis e T. forsythia, embora relativamente prevalentes nas infecções endodônticas, não apresentaram correlação significativa com o desenvolvimento de sintomatologia dolorosa. Por fim, a ausência de agrupamentos de perfis bacterianos quanto aos parâmetros sintomatológicos sugere que a estrutura das comunidades bacterianas intra-radiculares não possui influência significativa no desenvolvimento da dor de origem endodôntica
Abstract: The objective of the present study was to characterize the bacterial communities associated with endodontic infections by use of culture-independent molecular techniques. Overall, 34 intraradicular samples from teeth harboring endodontic infections were evaluated. 16S ribosomal DNA (16S rDNA) clone library analysis allowed the identification of 2 to 14 bacterial phylotypes (species) per tooth (mean= 9.6), with a total of 46 distinct phylotypes. Among the latter, 4 (9%) were considered previously unreported and further taxonomically classified as members of the order Clostridiales. Well-known endodontic representatives of Campylobacter, Eubacterium, Peptostreptococcus, Selenomonas, Treponema e Veillonella were detected, as well as members of less frequently reported genera, such as Burkholderia, Filifactor and Megasphaera. The application of the Real Time PCR technique permitted the detection of P. gingivalis, T. forsythia and a coexistence of both in 24%, 56% e 18% of the subjects, respectively. No significant correlations were evidenced among the levels of P. gingivalis and T. forsythia, individually or conjointly, and spontaneous endodontic pain. The use of T-RFLP in the analysis of bacterial community structures revealed a total of 123 (HhaI endonuclease) and 122 (MspI endonuclease) distinct terminal restriction fragments (T-RFs), with 20.8 and 20.0 mean T-RFs per tooth, respectively. Approximately 50% of the detected fragments were exclusive to one or two patients, indicating a high inter-subject variability in the bacterial assemblages. Cluster and multivariate statistical analyses did not demonstrate significant differences in the bacterial community profiles among the asymptomatic, tender to percussion and symptomatic study groups. Taken together, the results of this study reiterate the concept that the microbiota associated with endodontic infections is essentially polymicrobial, highly variable among individuals, and predominantly composed of Gram-positive anaerobic bacteria from the phylum Firmicutes. The species P. gingivalis and T. forsythia, although relatively prevalent in root canal infections, did not present significant correlations with the development of symptomatic features. Lastly, the absence of clusters of bacterial profiles according to symptomatic parameters suggests that the intraradicular bacterial community structures, as a whole, do not bear significant influence on the development of pain of endodontic origin
Doutorado
Microbiologia e Imunologia
Doutor em Biologia Buco-Dental
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Kirker, Grant Terral. "Effects of chlorothalonil (CTN) and butylated hydroxy-toluene (BHT) on microbial communities involved in the deterioration of wood using terminal restriction fragment length polymorphism (T-RFLP) analyses." Diss., Mississippi State : Mississippi State University, 2008. http://library.msstate.edu/etd/show.asp?etd=etd-04022008-155301.
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Peyret, Guzzon Marine. "Etudes moléculaires de la diversité des communautés et populations de champignons mycorhiziens à arbuscules (Glomeromycota)." Thesis, Dijon, 2014. http://www.theses.fr/2014DIJOS065/document.
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La symbiose mycorhizienne à arbuscules, dont l’apparition est conjointe à celle des plantes terrestres il y a 460 millions d’années, est une association mutualiste à bénéfices réciproques qui s’instaure entre la plupart des plantes terrestres, y compris celles cultivées, et des microorganismes ubiquitaires du sol que sont les champignons mycorhiziens à arbuscules (CMA, phylum des Glomeromycota). Lors cette symbiose, le fort potentiel d’amélioration de la nutrition minérale des plantes, et donc de la production végétale, est un atout dans le contexte mondial actuel d’augmentation de la demande de la production agricole. Afin d’optimiser les services écosystémiques des CMA dans les écosystèmes et en particulier les agroécosystèmes, la maîtrise de cette symbiose en ingénierie écologique nécessite la compréhension des mécanismes complexes qui régissent la dynamique de cette symbiose dans ces écosystèmes. Pour cela, nous avons étudié la diversité des communautés et des populations de CMA dans les agroécosystèmes à différentes échelles spatiales et sous l’influence de différentes pratiques culturales par des techniques d’empreintes moléculaires: séquençage haut-débit et polymorphisme de longueur de fragments de restriction. Les résultats obtenus montrent que la structuration de la diversité des CMA est influencée par le type d’usage de sol (prairie vs. culture), les pratiques culturales (retournement du sol, fertilisation et système de culture) ainsi que par les facteurs abiotiques (e.g. pH du sol). En conclusion, ces différents facteurs sont à prendre en compte dans l’optimisation des services écosystémiques des CMAThe arbuscular mycorrhizal symbiosis, which appeared at the same time as land plants, 460 million years ago, is a mutualistic beneficial association between most land plants, including those cultivated, and arbuscular mycorrhizal fungi (AMF). AMF, from the Glomeromycota phylum, are widespread soil microorganisms needing a photosynthetic host to complete their life cycle (obligate symbionts). The great potential of plant mineral nutrition improvement and crop production increased during this symbiosis, make AMF an asset in the context of an increase in the demand of world food crop production. The control of that symbiosis by ecology engineering in order to improve ecosystem services, especially in agroecosystems, needs to better understand the mechanisms regulating its dynamic. Therefore, we studied community and population diversity of AMF under influences of different agricultural practices at several spatial scales using genetic fingerprinting methods: high-throughput sequencing and restriction fragment length polymorphism. Results show that AMF diversity is structured by land use type (grassland vs. arable fields), cultural practices (soil disturbance, fertilizations, culturing systems) as well as environmental factors (e.g. soil pH). In conclusion, those different factors have to taken in account in AMF ecosystemic service managing
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Corniquel, Béatrice. "Construction de banques d'ADNc de palmier dattier (Phoenix dactylifera L. ). Différenciation de cultivars par RFLP à l'aide des ADNc : isolement et caractérisation d'un ADNc polymorphe. Isolement et caractérisation d'un ADNc codant pour la glutamine synthetase cytosolique." Angers, 1994. http://www.theses.fr/1994ANGE0014.
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La caractérisation des cultivars de palmier dattier a été réalisée par des analyses RFLP et RAPD à partir des feuilles internes des rejets. Des sondes ont été isolées d'une banque d’ADNc construite à partir d'ARNM extraits de cals organogènes du cultivar bou sthammi noire. La sonde ADNc 1 a permis d'obtenir neuf profils d'hybridation distincts pour chacun des neuf cultivars étudiés. Des profils d'amplification distincts pour trois cultivars ont été obtenus par RAPD avec des amorces oligonucléotidiques. Des analyses RFLP ont été aussi réalisées sur du matériel maintenu in vitro ou issu de culture tissulaire afin de déterminer si d'éventuelles variations induites par la culture in vitro affectaient le génome du palmier dattier. L'analyse moléculaire de la sonde ADNc 1 a été abordée. Le transcrit qui comprend 2400 nucléotides est exprimé dans les feuilles et les cals organogènes. Sa séquence partielle a été établie, l'absence d'homologie entre cette séquence de 1456 nucléotides et les séquences enregistrées dans les banques de données suggère que nous avons isole un ADNc codant pour une nouvelle protéine. Un ADNc codant pour la glutamine synthétase a été isolé. L'absence au niveau de la séquence des aminoacides de cet ADNc de l'extension c-terminale qui caractérise la structure primaire des séquences des sous-unités chloroplastiques montre que l’ADNc isole code pour la sous-unité cytosolique de la glutamine synthétase.
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Bouzid, Makhlouf. "Polymorphisme génétique du virus d'Epstein-Barr en Afrique du Nord : étude dans le carcinome du rhinopharynx, la maladie de Hodgkin et les lymphomes malins non-Hodgkiniens." Université Joseph Fourier (Grenoble), 1998. http://www.theses.fr/1998GRE10111.
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L'infection quasi mondiale de l'homme par le virus d'epstein-barr (ebv) contraste avec, la restriction geographique de certaines pathologies qui lui sont associees, l'incidence et la distribution en fonction de l'age variant pour une meme maladie a travers le monde. Cette situation complexe suggere l'existence de differentes souches d'ebv, specifiques de regions geographiques ou de certaines pathologies. Le carcinome du rhinopharynx (npc) est une des pathologies associees a l'ebv, qui se caracterise par une incidence intermediaire en afrique du nord en affectant deux tranches d'age de la population : la premiere de 10 a 24 ans et la deuxieme de 45 a 55 ans. La maladie de hodgkin (mdh) est une autre pathologie associee a l'ebv relativement frequente en afrique du nord, ou elle predomine chez l'enfant alors qu'elle affecte les adultes dans les pays industrialises. Nos resultats dans l'analyse genotypique du virus d'epstein-barr implique dans le npc, dans la mdh ou present dans la population saine ebv-positive, ont montre que la souche d'ebv associee au npc d'afrique du nord est de type a/f/wi/xho1kept/h1h2, donc completement differente de celle associee au npc asiatique, de type a/f/wi/xho1lost/h. La meme souche de npc algerien est retrouvee dans les deux pics d'age et associee aux differents stades cliniques de la maladie. Cependant un autre type d'infection est associe a la maladie de hodgkin, qui se caracterise : premierement, par une coinfection avec le type a et le type b d'ebv, et deuxiemement, par le genotype h. Nos resultats ont montre aussi qu'en algerie le genotype h est dominant dans la population generale, dans les lcls etablies avec le virus oropharynge de patients npc ou d'individus sains ebv-positifs, dans la salive de personnes atteintes de npc ou de mdh. L'ensemble de ces resultats soutient l'hypothese d'une specificite pathologique du virus d'epstein-barr en afrique du nord.
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Coelho, Andrea Gobetti Vieira. "Tuberculose multirresistente e extensivamente resistente em área metropolitana de elevada incidência - município de Santos (SP), Brasil." Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/5/5134/tde-19052015-143414/.
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INTRODUÇÃO: A incidência de tuberculose (TB) em Santos (SP) situa-se em 73/100.000 habitantes-ano. A prevalência média de coinfecção TB/HIV é de 16%, taxas de cura e abandono de tratamento, entre casos novos são, respectivamente, 71% e 12%. Tais indicadores sugerem elevado risco para TB multidroga resistente (TBMR) no município, com incidência estimada em 1,9/100.000 habitantes-ano. OBJETIVO: Descrever e analisar o perfil de sensibilidade às drogas (TS) de primeira e segunda linha de tratamento entre pacientes com TB pulmonar (TBP), estimar a incidência de TBMR e a proporção de TB extensivamente resistente (TBXR); analisar aspectos moleculares, epidemiológicos e institucionais dos casos de TBP resistentes em Santos (SP). MÉTODOS: Estudo descritivo de uma coorte de pacientes de TBP, com início de tratamento ou retratamento entre 01 de janeiro de 2011 a 31 de dezembro de 2012. Definiu-se como caso de TBP, indivíduos com 15 anos ou mais, de ambos os sexos, residentes no município de Santos, com manifestações clínicas compatíveis com TBP e confirmação por cultura com isolamento de Mycobacterium tuberculosis. As variáveis de interesse para o estudo foram as características sociodemográficas, história atual e pregressa de TB, aspectos relativos ao tratamento, co-morbidades, ao diagnóstico e resistência a drogas. Para as análises comparativas entre proporções foram usados os testes qui-quadrado de Pearson e o Exato de Fisher e para variáveis contínuas o teste T de Student ou o de Kruskal - Wallis. Os perfis genéticos dos isoladas resistentes a ao menos uma droga foram obtidos pela técnica RLFP e analisados pelo programa Bionumerics versão 5.0 (Applied Maths - Bélgica). A descrição da distribuição espacial da TB resistentes e clusters foram feitas mediante a inserção dos casos no mapa de Santos, por endereço de residência, segundo o índice de vulnerabilidade social. RESULTADOS: Dos 263 casos de TBP selecionados, 68,4% (180/263) eram do sexo masculino, a mediana da idade foi de 38 anos, 8,7% (23/263) eram diabéticos; 20,4% (42/206) dos casos novos apresentavam ao menos um fator de risco para TBMR; destacando-se entre estes casos 10,7% (22/206) de confecção HIV/TB; 47,3% (123/260) tiveram tratamento supervisionado, 14,7% (91/617) dos contatos foram examinados, 18,6% (49/263) foram hospitalizados durante o tratamento, perfazendo uma média de 145,4 dias por paciente. Entre os casos resistentes a ao menos uma droga, a resistência à isoniazida foi 8,4% (22/263) e à rifampicina 3,8% (10/263) dos casos. A TBMR primária foi encontrada em 1,9% (4/206) dos casos e destes 25,0% (1/4) eram TBXR. A incidência média anual de TBMR foi de 0,57/100,000 habitantes. Dos 25 isolados resistentes ao menos uma droga, submetidos à RFLP, 12 (48,0%) foram agrupados em seis grupos genéticos, com dois pacientes em cada grupo. CONCLUSÕES: A elevada proporção TBMR primária, com um caso de TBXR enfatizam a necessidade de universalizar a cultura e TS, ampliar a cobertura do tratamento supervisionado, a investigação rotineira dos contatos e o monitoramento da resistência a drogas. O fortalecimento da vigilância da resistência às drogas é indispensável para o contínuo aperfeiçoamento do Programa de Controle da TB, especialmente em regiões de elevada carga da doençaINTRODUCTION: The incidence of tuberculosis (TB) in Santos (SP) is located around 73 / 100,000-year, approximately double that found on average in the country. The average prevalence of TB / HIV is 16% cure rates and treatment dropout among new cases are, respectively, 71% and 12%. Such indicators suggest high risk for multidrug-resistant TB (MR-TB) in the city, with the incidence estimated at 1.9 / 100,000-year. OBJECTIVE: To describe and analyze the sensitivity to drugs of first and second line treatment of patients with pulmonary TB (PTB) to estimate the incidence of MR-TB and extensively drugresistant TB (TBXR), describe molecular and institutional aspects, spatial distribution, epidemiological PTB resistant cases in the city of Santos (SP). METHODS: A descriptive study of a cohort of patients with PTB residing in the city who started treatment or retreatment in the period January 2011 to December 31, 2012. The case definition PTB individuals 15 years or more, both sexes, living in the city of Santos (SP), who present clinical manifestations compatible with PTB and whose confirmation was made by culture with isolation of M. tuberculosis. The variables of interest for the study were: bacteriological / laboratory socio-demographic characteristics, current and previous history of TB, aspects related to treatment, and comorbidities. For comparative analyzes of proportions the chi-squared tests and Fisher\'s exact were used for continuous variables and the Student t test or the Kruskal - Wallis. The genetic profiles of isolates resistant to at least one drug were obtained by RFLP (length polymorphism restriction fragment) and analyzed using version BioNumerics 5.0 (Applied Maths - Belgium) software. The description of the spatial distribution of resistant TB and the clusters was made by inserting the cases in Santos map, by address of residence, which was according to the index of social vulnerability. RESULTS: Of the 263 cases of PTB selected, 68.4% (180/263) were male, th median age was 38 years, 8.7% (23/263) were diabetes; 20.4% (42/206) of new cases had at least one risk factor for MR-TB, especially 10.7% (22/206) of making HIV / TB; 47.3% (123/260) underwent supervised treatment, 14.7% (91/617) of the contacts were examined, 18.6% (49/263) were hospitalized during treatment, totaling 7127 days of hospitalization with a mean 145.4 days per patient. Among the cases resistant to at least one drug resistance to isoniazid 8.4% (22/263) and rifampin 3.8% (10/263) of the cases was found. The primary MR-TB was found in 1.9% (4/206) of MR-TB cases and of these 25.0% (1/4) were TBXR. The average annual incidence of MDR-TB was 0.57/100,000 inhabitants. Of the 25 isolates resistant least one drug, subjected to molecular characterization of IS6110, 12 (48.0%) were grouped in six clusters, with each group including two isolates. CONCLUSIONS: A high proportion of primary MR-TB, including a case of TBXR emphasizes the need to universalize culture and TS, expand the coverage of supervised treatment, routine investigation of contacts and monitoring of drug resistance. The strengthening of the surveillance of drug resistance is essential for continuous improvement of the TB Control Program, especially in regions of high disease burden
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Thomas, Martine. "Apport de la biologie moléculaire à l'épidémiologie et la physiopathologie de l'infection chlamydienne." Compiègne, 1995. http://www.theses.fr/1995COMP862S.
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The Polymerase Chain Reaction technic was applied to the epidemiological and immunopathological study of Chlamydia trachomatis genital infections. Compared to a reference method combining cell culture and PCR, the PCR technic shows a sensitivity of 93. 6% and a specificity of 83. 8%, while cell culture presents a sensitivity of 47. 6% and a specificity of 98%. One hundred and seventy nine clinical isolates of C. Trachomatis were serotyped using the microimmunofluorescence method and genotyped by Restriction Fragment Length Polymorphism. One hundred and forty eight strains (82. 7%) gave similar results by the two technics, but 31 strains (17. 3%) gave discrepant results. These discrepancies are explained by a lack of specificity of the monoclonal antibodies. The epidemiological results obtained by RFLP are in agreement with the results observed in different countries at different time points. In order to find new epidemiological markers, we demonstrated the presence of repetitive extragenic palindromic sequences (REP) in C. Trachomatis chromosome. The Rep-PCR amplification pattern of C. Trachomatis differs from the amplification patterns of other bacteria. The PCR was used to detect cytokines mRNA in the genital tracts of mice infected with a human strain of C. Trachomatis. We detected an increase in the transcription of IL-1a, IL-1b, IL-2, IL-6, IL-10, IL-12, TNFa, IFNg and Macrophage Inflammatory Protein genes at D4 post-infection in infected mice compared to normal mice. Transcription rates of IL-2, IL-12 and IFNg remain high at D14 while the transcription rates of the other cytokines return to a baseline level. Therefore, C. Trachomatis infection seems to induce a Th1 immune response in vivo in a mouse model.
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Gunnar, Erika. "Characterization of the genetic basis in two cases of abetalipoproteinemia reveals two novel mutations." Thesis, Linköping University, Department of Physics, Chemistry and Biology, 2010. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-58620.
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BACKGROUND: Abetalipoproteinemia (ABL) is a rare autosomal recessive disorder caused by mutations in the gene coding for microsomal triglyceride transfer protein (MTTP).
AIM: To characterize the genetic basis of ABL in two unrelated patients.
RESULTS: In the first patient, the substitution c.1911C>T in exon 12 of the MTTP gene, resulting in the protein substitution p.P552L, was discovered using mutation screening. The parents are heterozygous and the proband is a homozygous carrier of this substitution. Using restriction fragment length polymorphism (RFLP), 100 control subjects were analyzed and none carried the substitution indicating that it is a novel MTTP mutation. Sequencing of the other ABL patient showed that the proband carried a homozygous single base insertion, at position c.2342IVS16+2-3insT, located at the donor splice-site of intron 16 resulting in skipping of exon 16 and truncation of the protein. The proband's mother is heterozygous for the insertion while the father does not carry the insertion. Multiplex ligation-dependent probe amplification (MLPA) did not identify any deletion encompassing exon 16 in the proband, father or mother. Nonpaternity was excluded using polymorphic markers from several chromosomes. Haplotype analysis using markers spanning chromosome 4 revealed heterodisomy (two homologous chromosomes) of 4p and the distal part of 4q, and isodisomy (duplication of one chromosome) of 4q12-4q26.
CONCLUSION: These data show that the cause of ABL in one of the patients is a missense mutation, p.P552L, while the cause of ABL in the other patient is due to uniparental disomy, probably resulting from non-disjunstion in meiosis I.
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Marrot, Laurent. "Mise en evidence du polymorphisme de l'adn a l'aide de sondes chimiques." Orléans, 1988. http://www.theses.fr/1988ORLE2010.
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Le polymorphisme de l'adn et les distorsions induites par la fixation d'un agent antitumoral: le cis-ddp sont etudies a l'aide de reactifs specifiques de la nature et de la conformation des bases. Le bromo ou chloroacetaldehyde permet de cartographier les adenines et les cytosines d'une sequence en conformation z et des jonctions adn-b-adn-z. La caracterisation des distorsions induites par le cis-ddp sur des oligonucleotides et sur les fragments d'adn a ete realisee a l'aide de sondes chimiques. L'accessibilite de la thymine complementaire de l'adenine d'un adduit d(apg) depend de la sequence environnante, la cytosine complementaire n'est pas accessible
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Henrion, Bénédicte. "Caractérisation et identification de champignons ectomycorhiziens par amplification enzymatique (PCR) de l'ADN ribosomal : application au suivi du basidiomycète laccaria bicolor en pépinière forestière." Nancy 1, 1993. http://www.theses.fr/1993NAN10329.
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Afin d'identifier des champignons mycorhiziens dans les écosystèmes forestiers, à différents niveaux taxonomiques, un diagnostic moléculaire basé sur le polymorphisme de l'ADNr a été mis au point. Le polymorphisme de l'ADN ribosomal, étudie par PCR couplée à la RFLP, des régions codantes et non codantes, permet la caractérisation au niveau du genre, de l'espèce ou de l'isolat d'un grand nombre de champignons ectomycorhiziens. Cette identification est réalisée à partir de mycélium, de carpophores et de mycorhizes. Ce diagnostic moléculaire est utilisé en épidémiologie afin d'étudier la structure et la dynamique des communautés mycorhiziennes. Il a permis d'apprécier la survie et la compétitivité du champignon ectomycorhizien, laccaria bicolor s238, par l'identification de ces ectomycorhizes, dans une expérimentation en pépinière forestière
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Chikaha, Tchouya-Ngandjio Antoinette Brigitte. "Epidémiologie des infections urogénitales à Chlamydia trachomatis chez des étudiants camerounais (Yaoundé) : génotypage des souches, sensibilité aux antibiotiques : mise au point de la détection quantitative par PCR en temps réel." Paris 6, 2003. http://www.theses.fr/2003PA066055.
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Léveillard, Thierry. "Le polymorphisme des gènes de l'inter-alpha-trypsine inhibiteur : recherche d'association génétique avec l'emphysème pulmonaire." Rouen, 1989. http://www.theses.fr/1989ROUES015.
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RFLP des gènes de l'inhibiteur de la trypsine inter alpha chez l'homme, déterminisme génétique et fréquence allélique de ces marqueurs dans une population de référence et dans une population constituée d'individus non déficitaires en alpha-1-antitrypsine souffrant d'emphysème pulmonaire
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Corrêa, Daniele Bussioli Alves 1985. "Caracterização morfológica, patogênica e molecular de linhagens de Streptomyces associadas à sarna da batata de diferentes regiões produtoras do Brasil." [s.n.], 2011. http://repositorio.unicamp.br/jspui/handle/REPOSIP/317453.
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Orientador: Suzete Aparecida Lanza DestéfanoDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
Made available in DSpace on 2018-08-17T18:58:40Z (GMT). No. of bitstreams: 1 Correa_DanieleBussioliAlves_M.pdf: 2704976 bytes, checksum: 40df9160618a30b2959f8df5ff93e585 (MD5) Previous issue date: 2011
Resumo: A batata ocupa o quarto lugar em volume de produção mundial de alimentos após o arroz, trigo e milho. O Brasil é o maior produtor dentre os países da América Latina, porém ainda apresenta baixa produtividade devida às doenças que afetam a cultura. Dentre as doenças bacterianas, a sarna da batata é uma das mais importantes economicamente e sua ocorrência é generalizada no mundo. Diferentes espécies do gênero Streptomyces estão associadas à essa doença e os principais sintomas se caracterizam por lesões irregulares que podem tomar toda a superfície do tubérculo, acarretando diminuição do seu valor comercial ou, até mesmo, impedindo a sua comercialização. Atualmente, a incidência da sarna está aumentando consideravelmente, tornando-se um fator limitante do cultivo de batata no Brasil. O presente trabalho teve por objetivo a identificação de linhagens de Streptomyces sp. associadas à sarna da batata, provenientes de diferentes regiões produtoras do Brasil, por meio de taxonomia polifásica. Cento e noventa linhagens de Streptomyces foram analisadas, sendo 165 nacionais obtidas dos estados da Bahia, Goiás, Minas Gerais, Paraná, São Paulo, Rio Grande do Sul e Santa Catarina; 13 de material vegetal importado do Chile, França e Holanda; e 12 linhagens Tipo de Streptomyces representantes de espécies associadas à sarna. Na caracterização morfológica as linhagens apresentaram heterogeneidade com relação à micromorfologia de hifas e coloração dos esporos. A patogenicidade das linhagens foi avaliada pela presença dos genes nec1, tomA (tomatinase) e txtAB (taxtomina A) e os resultados indicaram que 94 linhagens (52,8%) apresentaram amplificação dos três genes de patogenicidade avaliados, 14 (7,9%) não apresentaram nenhum dos genes e 70 (39,3%) mostraram sinal positivo de amplificação para um ou mais genes. Para algumas linhagens a confirmação da patogenicidade foi efetuada por meio de testes em minitubérculos de batata. Nos testes moleculares, incluindo amplificação com primers específicos para S. scabiei e S. turgidiscabies, PCR-RFLP do gene atpD e análises das seqüências dos genes atpD, gyrB, recA, rpoB e trpB, foi possível a separação das linhagens analisadas em diferentes perfis genéticos. As análises das características morfológicas, patogênicas e moleculares permitiram a identificação de 57 linhagens pertencentes à S. scabiei, 28 à espécie S. ipomoeae, 13 à S. caviscabies/S. setonii, 12 à S. europaeiscabiei e duas à S. sampsonii. As 66 linhagens restantes apresentaram características distintas das espécies Tipo testadas, podendo representar possíveis novas espécies de Streptomyces associadas à sarna no Brasil
Abstract: Potato is the world's fourth most important food crop after rice, wheat and maize. Brazil is the biggest producer among the countries of Latin America, but it still has low productivity due to diseases that affect the crop. Among the bacterial diseases, the potato scab is one of the most economically important, and its occurrence is widespread in the world. Different species of the genus Streptomyces are associated with this disease and the main symptoms are characterized by irregular lesions that can affect all the tuber surface causing decrease of its commercial value or preventing its commercialization. Currently, the incidence of potato scab is increasing considerably, becoming a limiting factor in potato production in Brazil. This study aimed to identify Streptomyces strains associated with potato scab, from different potato growing areas in Brazil, through polyphasic taxonomy. One hundred and ninety strains of Streptomyces were analyzed, including 165 Brazilian strains obtained from potatoes coming from the states of Bahia, Goias, Minas Gerais, Parana, Sao Paulo, Rio Grande do Sul and Santa Catarina; 13 of plant material from Chile, France and Netherlands; and 12 type strains of Streptomyces species associated with potato scab. In the morphological characterization, the strains showed heterogeneity in the hyphal micromorphology and color of spores. The pathogenicity of strains was investigated by presence of the nec1 and tomA genes, and txtAB operon (thaxtomin A). The results indicated that 94 strains (52.8%) showed amplification of the three pathogenicity genes, 14 (7.9%) showed no amplification of the genes and 70 (39.3%) showed positive signal for only one or more genes. The pathogenicity of some strains was confirmed with artificial inoculation onto potato minitubers. In the molecular tests, including PCR amplification with specific primers for S.scabiei and S. turgidiscabies, analysis of PCR-RFLP of atpD gene and sequences analysis of the atpD, gyrB, recA, rpoB and trpB genes, the strains could be separated into different genetic profiles. The morphological, pathogenic and molecular data allowed identifying of 57 strains belonging to S. scabiei, 28 to S. ipomoeae, 13 to S. caviscabies/S. setonii, 12 to S. europaeiscabiei and two to S. sampsonii. The 66 remaining strains showed different genetic profiles in comparison with the type strains of Streptomyces, and may represent new species of Streptomyces associated with potato scab in Brazil
Mestrado
Genetica de Microorganismos
Mestre em Genética e Biologia Molecular
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Moreira, Abdiel Aparecido. ""Pesquisa de sítios de restrição enzimática em segmento da ORF K1 do genoma de herpesvírus humano tipo 8 (HHV-8) em isolados clínicos de São Paulo: relação com subtipos virais e implantação da técnica de RFLP (Restriction Fragment Length Polymorphism Analyses) para determinar subtipos virais"." Universidade de São Paulo, 2003. http://www.teses.usp.br/teses/disponiveis/9/9136/tde-01102004-164856/.
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A epidemia da Síndrome de Imunodeficiência Adquirida (AIDS) fez aumentar a incidência de sarcoma de Kaposi (SK) em todos os países e o SK passou a ser considerado doença definidora de AIDS. Desde a descoberta de seu agente etiológico, o herpesvírus humano tipo 8 (HHV-8), vários estudos vêm sendo realizados com o objetivo de caracterizar subtipos virais presentes em todas as formas de SK: clássica, endêmica, iatrogênica e epidêmica. Os sistemas rotineiramente usados na subtipagem do HHV-8 têm usado o seqüenciamento de um gene que contém regiões hipervariáveis e que codifica uma glicoproteína de membrana viral (ORF K1). O presente trabalho apresenta um sistema de subtipagem alternativo que se baseia na presença de sítios de restrição enzimática em um pequeno segmento do gene ORF K1, região hipervariável 1 (VR1) e que permite discriminar subtipos virais. A análise de 68 seqüências; 50 que pertenciam a 36 pacientes com sarcoma de Kaposi infectados e não infectados pelo HIV-1 de São Paulo e 18 a protótipos dos subtipos A a E, mostraram mapas de restrição enzimática característicos dos principais subtipos virais descritos até o momento. Tomando como base apenas as enzimas disponíveis comercialmente, foram selecionadas cinco que se mostraram úteis para a subtipagem de HHV-8: Taq I, Nsi I, Hinf I, Hae III e Mse I. Os resultados obtidos com a técnica de PCR-RFLP (reação em cadeia de polimerase associada à análise do polimorfismo de fragmentos de restrição enzimática) mostraram que de 48 espécimes brasileiros previamente classificados como sendo dos subtipos A, B e C por seqüenciamento gênico, todos foram corretamente subtipados pela técnica de PCR-RFLP. Três amostras (duas do subtipo A e uma do B) apresentaram mais um sítio de restrição enzimática além dos descritos como sendo os predominantes. Mais recentemente, outras 27 amostras de 18 casos de infecção por HHV-8 foram subtipadas pela PCR-RFLP. Houve detecção de oito isolados do subtipo A, sendo seis de variante predominante, um de variante minoritária conhecida e um de nova variante viral. Dois casos de infecção por HHV-8 do subtipo B e sete do subtipo C também foram identificados. Finalmente, um provável caso de infecção pelo subtipo E foi encontrado em paciente com SK-AIDS disseminado, resistente à quimioterapia. Na avaliação global, houve maior número de casos de infecção por HHV-8 dos subtipos A e C. Concluindo, devido à alta sensibilidade e especificidade, baixo custo, rapidez e facilidade de execução, a técnica de PCR-RFLP pode ser usada em larga escala para estudos de epidemiologia molecular, principalmente em países em desenvolvimento.AIDS epidemic has increased the incidence rates of Kaposi s sarcoma (KS) in all countries, and KS has been considered an AIDS-defining illness. Since the discovery of the human herpesvirus 8 (HHV-8), the etiological agent of KS, several studies have been conducted in order to characterize HHV-8 in all forms of KS: classic, endemic, iatrogenic, and epidemic. The HHV-8 genome presents a hypervariable region termed ORF K1 useful for virus subtyping. The objectives of the present study were to describe an alternative method for subtyping HHV-8, to compare this new method with DNA sequencing, and to use this method for HHV-8 subtyping. After cloning and sequencing a segment of the ORF K1 (VR1) in 50 HHV-8/DNA isolates from 36 Brazilian KS-AIDS patients, we searched for restriction enzymatic sites in this segment of DNA, and compared them with 18 sequences reported in the literature. Then we constructed the enzymatic restriction maps useful for discriminating all HHV-8 subtypes described up to now, and standardized a PCR-RFLP (restriction fragment length polymorphism analysis) using five commercial enzymes: Taq I, Nsi I, Hinf I, Hae III e Mse I. After comparing the results obtained by the two methods, we used PCR-RFLP for HHV-8 subtyping in 27 new HHV-8/DNA isolates. The results obtained by DNA sequencing and PCR-RFLP showed 100% of concordance, and allowed the use of PCR-RFLP for HHV-8 subtyping. Indeed, we disclosed that among KS-AIDS patients from São Paulo, subtypes A and C are more prevalent than subtype B. Although additional restriction sites were detected in some Brazilian HHV-8 isolates, the majority of them belonged to the predominant strains described in the literature. Interestingly, one probable case of HHV-8 subtype E was detected in a patient who presented disseminated KS and resistance to chemotherapy. Because of its high sensitivity, specificity, low cost, and rapid execution, PCR-RFLP could be used on a large scale, mostly in countries with poor resources and where KS is endemic.
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Watson, Brian T. "Population biology and fish hosts of several federally endangered freshwater mussels (Bivalvia: Unionidae) of the upper Tennessee River drainage, Virginia and Tennessee." Thesis, This resource online, 1999. http://scholar.lib.vt.edu/theses/available/etd-08222008-063606/.
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Chausse, Anne-Marie. "Approche moleculaire de l'association entre le complexe majeur d'histocompatibilite et les maladies dans deux especes differentes." Paris 7, 1988. http://www.theses.fr/1988PA077033.
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Wintz, Henri. "Contribution a l'etude de l'organisation et de la structure des genes de rna de transfert mitochondriaux des plantes." Université Louis Pasteur (Strasbourg) (1971-2008), 1988. http://www.theses.fr/1988STR13008.
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Les genes codant pour le trna**(phe), trna**(trp), trna**(typ), trna**(met), trna**(pro), trna**(asp) et trna**(his) ont ete identifies a l'aide de trnas isoaccepteurs purifies et d'oligonucleotide de synthese. Localisation des genes de trna chloroplastiques presents dans le genome mitochondrial de mais. Determination de la sequence nucleotidique de deux genes de trna (trna**(cys) et trna**(ser)) et d'un pseudo-gene de trna**(pne) inactive par la presence d'une insertion dans la sequence codante du gene. Le gene de trna**(cys) fait partie d'une insertion de pna chloroplastique dans le genome mitochondrial. Mise en evidence de deux phases de lecture ouverte susceptible de coder pour une sous unite(n**(o) 3) de la nadh dehydrogenase et pour la proteine s12 de la petite sous unite du ribosome. L'etude comparative de ces genes suggere que ces genes mitochondriaux et chloroplastiques ont une origine commune
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Amorim, Paulo Vinícius Gonçalves Holanda. "Análise da expressão do KISS1/KISS1 do polimorfismo rs5780218 KISS1 em somatotropinomas e adenomas hipofisários clinicamente não funcionantes." Universidade de São Paulo, 2018. http://www.teses.usp.br/teses/disponiveis/5/5135/tde-02082018-121901/.
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Apesar de amplamente estudados, os mecanismos envolvidos no processo de transformação neoplásica das células hipofisárias e na progressão desses tumores permanecem, ainda, pouco esclarecidos. Os genes da kisspeptina (KISS1), originalmente identificado como um supressor tumoral, e de seu receptor (KISS1R) desempenham um papel crucial no eixo hipotalâmico-hipofisário-gonadal e sua perda de expressão foi, recentemente, relacionada ao surgimento dos adenomas hipofisários. Com o objetivo de estudar a importância do KISS1/KISSR nesses tumores, foram avaliadas a expressão desses genes e a frequência do polimorfismo rs5780218 na região promotora do KISS1, em somatotropinomas e adenomas hipofisários clinicamente não funcionantes (ACNF). Foram avaliados 97pacientes, 49 somatotropinomas e 48 ACNF. DNA tumoral foi obtido de todos os tumores e RNA de 68 caso. Desses tumores, 52 foram classificados como invasivos (44 apresentavam invasão apenas para o seio cavernoso) e 45 não invasivos. A quantificação da expressão do mRNA de KISS1 e KISS1R foi realizada pela qPCR em tempo real com sondas TaqMan utilizando o método de quantificação relativa 2-deltaCt. A determinação do genótipo do rs5780218 foi feita por PCR seguida pela técnica de polimorfismo no comprimento do fragmento de restrição (RFLP). O gene KISS1 apresentou-se hipo-expresso na vasta maioria dos pacientes avaliados (97.2% dos somatotropinomas e 92.6% do ACNF). Em relação ao KISS1R, este também estava hipo-expresso na maior parte dos pacientes (100% dos somatotropinomas e 84.4% dos ACNF). Hiperexpressão do KISS1 e KISS1R foi rara em ambos os subtipos tumorais. Em relação as características clínicas dos pacientes e ao fenótipo tumoral, como tamanho e invasividade, não foram encontradas diferenças significantes na expressão desses genes. A avaliação do polimorfismo rs5780218 no KISS1 mostrou que o genótipo homozigoto para o alelo variante foi bem mais frequente nos somatotropinomas (32.6%) versus ACNF (10.4%; P=0.03). A presença do rs5780218 foi relacionada a invasividade tumoral, quando considerado apenas a invasão para o seio cavernoso (P=0.03). Esse é um dado interessante, já que a KISS1 pode formar um complexo com as metaloproteinases e estas têm sido relacionadas a invasão dos adenomas hipofisários para o seio cavernoso e não para o seio esfenoidal. Entre os pacientes com ACNF, o alelo variante foi mais frequente nos indivíduos do sexo masculino (P=0.02) e foi relacionado com uma menor idade de diagnóstico (mediana 33.0 anos, min 26 - max 42) quanto presente em homozigose (P < 0.01); os pacientes homozigotos para o alelo ancestral e heterozigotos apresentaram idade diagnóstica com mediana de 50.0 (min 19 - max 73) e 54.8 (min 17- max 84), respectivamente. Curiosamente, a expressão do KISS1 foi menor nos tumores com homozigose para o alelo mutante tantos nos somatotropinomas quanto nos ACNF. Em conclusão, foi observada hipo-expressão do KISS1 e KISS1R nos somatotropinomas e ACNF, podendo essa perda expressão estar relacionada a gênese desses adenomas. O alelo variante rs5780218 KISS1 foi relacionado a invasão para o seio cavernoso e encontrado em maior frequência nos somatotropinomas, sugerindo que a importância dos KISS1/KISS1R pode diferir entre os subtipos de tumores hipofisáriosAlthough broadly studied, the mechanisms involved in the neoplastic process of pituitary cells remains still unclear. Kisspeptin1 (KISS1), originally identified as a tumor suppressor, and its receptor (KISS1R) play a crucial role in the hypothalamic-pituitary-gonadal axis and the loss of their expression was, recently, associated with pituitary adenomas onset. Aiming to investigated the importance of KISS1/KISS1R in these tumors, expression of KISS1 and KISS1R was determined in somatotropinomas and nonfunctioning pituitary adenomas (NFPA). The frequency of the rs5780218 polymorphism, located in the KISS1 promoter region, was also evaluated. A total of 97 patients were assessed, 49 somatotropinomas and 48 NFPA. Among these, 52 were categorized as invasive (44 of which only showed invasion to the cavernous sinus). KISS1 and KISS1R mRNA expression was performed through RT-qPCR using TaqMan probes and the 2-deltaCt relative quantification method. KISS1 rs5780218 genotyping was done by PCR and restriction fragment length polymorphism (RFLP) method. The KISS1 gene was underexpressed in the vast majority of the cases (97.2% of the somatotropinomas and 92.6% of NFPA). KISS1Runderexpression have also been observed in most cases (100% of the somatotropinomas and 84.4% of the NFPA). KISS1 and KISS1R overexpression was rarely detected. No significant differences were found between KISS1 and KISS1R gene expression and patient\'s clinical characteristics and tumor phenotype, such as size and invasiveness. The characterization of rs5780218 showed that the variant genotype in homozygosis was much more frequent in somatotropinomas (32.6%) versus NFPA (10.4%; P=0.03). The presence of variant allele was associated with tumor invasiveness when considered invasion to the cavernous sinus only (P=0.03). This data is particularly interesting, since KISS1 has the ability for form a complex with metalloproteinases and these, for instance, are related to invasiveness of pituitary adenomas to the cavernous, but not to the sphenoidal, sinus. When considered only NFPA, the variant allele was more frequent in males (P=0.02) and was associated with earlier age at presentation (median 33 years old, min 26 - max 42) when in homozygosis (P < 0.01); the wilt-type homozygotes and heterozygotes had medians of 50.0 (min 19 - max 73) and 54.8 (min 17 - max 84), respectively. Curiously, KISS1 expression was lower in rs5780218 homozygotes both in somatotropinomas and NFPA. In conclusion, we have identified the KISS1 and KISS1R underexpression in both somatotropinomas and NFPA, which lead to notion that the loss of expression might be related to the genesis of these adenomas. The rs5780218 KISS1 variant allele was associated with invasion to the cavernous sinus and was found to be more frequent in somatotropinomas, suggesting that role of KISS1/KISS1R in tumor behavior might differ between pituitary adenomas subtypes
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Cadima, Bruno Ferencz Papp. "Determinação de polimorfismos dos genes ABCC2 e ABCG2 como fator preditivo de resposta ao tratamento com cisplatina em pacientes com carcinoma epidermóide de cabeça e pescoço." Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/5/5155/tde-27092010-160453/.
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Os transportadores da família ABC são proteínas transmembrânicas envolvidas com o tráfego de substâncias endógenas e exógenas do meio intracelular para o extracelular, sendo alvos de estudo na resistência celular a agentes quimioterápicos. O ABCC2 é um transportador transmembrânico que exporta ativamente fármacos aniônicos conjugados e facilita o transporte de agentes anticâncer. O ABCG2 é outro transportador transmembrânico que tem influência na farmacocinética e farmacodinâmica de certos xenobióticos e substratos endógenos; além disso, acredita-se que este gene contribui para a resistência a várias drogas. Por esses motivos identificamos por sequenciamento ou PCR-RFLP os polimorfismos dos genes ABCC2 (Val417Ile, Ser789Phe e Ala1450Thr) e ABCG2 (Val12Met, Gly126stop códon, Gly141Lys) em 90 pacientes portadores de carcinoma epidermóide de cabeça e pescoço (HNSCC) e tentamos correlacionar a presença do polimorfismo com resposta a tratamento que incluiu cisplatina em todos os pacientes. Não encontramos nenhuma correlação entre a presença de polimorfismo para Val12Met, Gly141Lys e Val417Ile, determinados em 68 pacientes tratados exclusivamente com cisplatina e radioterapia, e a resposta ao tratamento. As curvas de sobrevida, determinadas por Kaplan-Meier, considerando polimórfico os pacientes que continham pelo menos um dos alelos alterados e selvagem os que não tivessem nenhum deles, mostraram que os pacientes selvagens para o polimorfismo Val12Met tiveram tendência a uma pior sobrevida (sobrevida mediana de 18,7 meses) em relação aos pacientes polimórficos (sobrevida mediana não atingida; P=0,089 Teste de log-rank), já os pacientes selvagens para o polimorfismo Gly141Lys tiveram tendência a uma pior sobrevida (sobrevida mediana de 15,8 meses) em relação aos pacientes polimórficos (sobrevida mediana de 25,6 meses; P = 0,16 Teste de logrank). Não observamos correlação entre os outros polimorfismos e sobrevida. Quanto ao GLY126stop, somente um paciente foi identificado como polimórfico. Conclusões: No nosso estudo, a freqüência do polimorfismo Val12Met de 10% está próxima dos 18% descrito na população normal (Kobayashi 2004). Nosso trabalho foi o primeiro a correlacionar estes polimorfismos com resposta ao tratamento em pacientes com carcinoma epidermóide de cabeça e pescoço e indica que Val12Met e Gly141Lys e o gene ABCG2 como um todo são candidatos a um estudo maiorATP binding cassette (ABC) transporters form one of the largest transmembrane protein families. These proteins use cellular ATP to drive the transport of various substrates across cell membranes including many exogenous and endogenous compounds, which includes drugs used in cancer treatment. ABCC2 is an ATP binding cassette transporter which accepts a diverse range of substrates, including glutathione, glucuronide, and sulfate conjugates of many metabolites and xenobiotics. ABCG2 is a member of the ATP binding cassette (ABC) transporters whose function is to pump out of the cell a wide variety of endogenous and exogenous compounds. Widely expressed in stem cells, ABCG2 is also recognized as a universal marker of stem cells. For these reasons we had identified the following polymorphisms of ABCC2 gene: -Val417Ile, Ser789Phe and Ala1450Thr- and of ABCG2 gene as well: -Val12Met, Gly126stop códon, Gly141Lys in 88 patients with head and neck squamous cell carcinoma (HNSCC). Methodology included PCR - RFLP and direct sequencing. Survival analysis was done using Kaplan-Meier curves and response measured by RECIST criteria. Comparisons were done between polymorphic patients in which at least one polymorphism was present as opposed to the patients without the polymorphism. Correlation with response to treatment was studied for Val12Met, Gly141Lys e Val417Il in 68 patients exclusively treated with concomitant cisplatin and radiotherapy and no correlation was found between these markers and treatment response. Patients without the Val12Met presented a trend towards shorter survival (median survival 18.7 months) as compared to polymorphic patients (median survival not reached, long rank p= 0.089). Although statistical significance was not reached, patients wild type for Gly141Lys polymorphism (median survival 15.8 months) had shorter survival than polymorphic patients (25.6 months, p=0.16). We did not observe any other correlation between other polymorphisms and survival. With respect to Gly126stop, only one patient was identified as polymorphic and survival analysis was not possible. As far as we know this is the first study to try to correlate these polymorphisms with treatment response and survival in HNSCC patients. Although we were unable to draw any definitive conclusions, our results indicate that Val12Met and Gly141Lys deserve to be further studied in the future.
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Moisan, Jean-Paul. "Etude et caracterisation de marqueurs genetiques specifiques du chromosome x humain (suivi de) etude des rearrangements du recepteur a l'antigene, sur des lymphocites t actives in vivo." Université Louis Pasteur (Strasbourg) (1971-2008), 1987. http://www.theses.fr/1987STR13231.
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Grandjean, Frédéric. "Variabilité morphométrique et génétique chez les populations de l'écrevisse à pattes blanches Austropotamobius pallipes : implications biogéographiques." Poitiers, 1997. http://www.theses.fr/1997POIT2258.
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La variabilité interspécifique d'austropotamobius pallipes a été estimée par analyse de caractères morphométriques et par une étude du polymorphisme de l'adn mitochondrial (restriction fragment length polymorphism). En Poitou-Charentes, l'étude de la variabilité morphologique a révélé une différenciation entre la population des mares du Pinail et un groupe homogène compose de l'ensemble de huit autres populations issues de cours d'eau. Deux hypothèses ont été émises pour expliquer cette discrimination : la première indique une introduction à partir d'animaux issus d'une autre région et la seconde fait intervenir la notion d'effet de fondation. L'étude génétique a révélé l'existence de trois haplotypes dont deux sont distribués exclusivement dans les populations issues de cours d'eau. Le troisième est exclusivement recensé chez les animaux du Pinail. L'étude génétique a été étendue à l'ensemble de l'aire de repartition (européenne) de cette espèce qui comprend trois sous-espèces géographiquement distinctes. Les 6 types mitochondriaux détectés, se répartissent en trois groupes majeurs correspondant aux trois sous-espèces décrites chez a. Pallipes : a. P. Pallipes, a. P. Italicus et a. P. Lusitanicus. Les forts pourcentages de divergence nucléotidiques révèles entre les haplotypes caractérisant les trois groupes suggèrent une séparation très ancienne de ceux-ci estimée à environ 6 a 8 millions d'années (ma) si l'on considère un pourcentage de divergence de l'adn mt de 2% par ma. L'existence de trois zones de refuges (péninsule ibérique, sud de la France et les Balkans) ayant abritées chacun de ces groupes lors des périodes de glaciations du quaternaire peut être avancée. Enfin, une étude préliminaire de la variabilité génétique des populations française a révélé l'existence des trois sous-espèces au sein du réseau hydrographique français, résultant probablement de transferts de populations
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Baud, Sylvie. "Recherche de corrélations entre marqueurs moléculaires de type RFLP et caractères agronomiques chez Zea mays." Lyon 1, 1992. http://www.theses.fr/1992LYO10119.
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Dans le cadre d'un programme eureka, 170 marqueurs ont ete etudies a l'aide de l'outil rflp (restriction fragment length polymorphism) sur 35 lignees de mais. Les distances genetiques existant entre ces lignees, ont pu etre ainsi evaluees. La precision de l'outil, sa bonne representation de la variabilite existante entre les differents groupes genetiques du mais ont pu etre mis en evidence. Trois approches differentes de recherche de qtl (quantitative trait loci) ont, au prealable, ete comparees, a l'aide de donnees simulees: basees sur la regression phenotype sur genotype et considerant un ou deux marqueurs (anova et mapmaker-qtl) ou basee sur un modele non lineaire, considerant un ou deux intervalles consecutifs de marqueurs adjacents (qtlstat). Une carte etablie a partir de 68 marqueurs analyses sur la descendance f2 du croisement choisi a permis de localiser differentes regions chromosomiques impliquees dans l'expression de divers caracteres: des genes majeurs impliques dans des pigmentations, dans differentes resistances et des qtl lies au rendement, a la precocite. Ces differentes descendances ont ete analysees dans plusieurs lieux d'essais. Des regions chromosomiques liees a certains caracteres se sont averees communes a toutes les descendances observees dans les differents lieux. D'autres, par contre, sont apparues propres a une descendance particuliere observee dans un lieu donne: cette etude a confirme la sensibilite des qtl a l'environnement et la necessite, en recherche de qtl, de pouvoir tenir compte des interactions genotype x environnement
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Lirette, Nicole Therésè. "Carte de restriction et clonage de certains fragments de l'ADN mitochondrial de la palourde de dune Spisula solidissima." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp03/MQ37853.pdf.
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