Dentro do gênero Adesmia, as técnicas moleculares ainda não foram empregadas na caracterização de germoplasma e na análise da diversidade genética das espécies brasileiras que compôem o gênero. Portanto os objetivos deste trabalho foram: caracterizar, com a utilização de marcador molecular do tipo RAPD, as espécies brasileiras do gênero Adesmia DC; com base nestas informações estabelecer relações de diversidade genética entre as espécies e os acessos analisados; relacionar dados de diversidade com dados morfológicos e de reprodução.

Made available in DSpace on 2016-06-02T19:29:41Z (GMT). No. of bitstreams: 1 TeseAJM.pdf: 1132829 bytes, checksum: d7a4ea748d124c0e0dddf18933cbad53 (MD5) Previous issue date: 2003-04-29 / Universidade Estadual de Maringá / Genetic and chemical variability of Maytenus ilicifolia Maytenus ilicifolia is a native plant of Southern Brazil commonly used as a popular medicine for indigestion, gastritis and ulcers. The large use of this plant has increased the degradation of the species, and thus it is presently included in FAO´s list for priority species for studying and conservation in South America. In this context, this work aimed to contribute with studies that help the conservation of this species. Initially, a survey in herbaria and in the field was performed to identify the incidence and distribution of the genus Maytenus in the State of Rio Grande do Sul. Three new species (M. glaucenscens Reiss, M. gonoclada Mart. and M. robusta Reiss) were found in this State. The genetic variability study of three populations showed that the intrapopulational variation (variance from 0.102 to 0.140) is higher than the interpopulational (variance between 0.076 and 0.099%). The three populations analyzed grouped themselves with low significance level and 7.6% of the present alleles are rare. The genetic variability study involving 18 native populations of Maytenus ilicifolia, Maytenus aquifolia and Maytenus evonymoidis demonstrated that the species may be separated with 100% of confidence, and that M. ilicifolia and M. aquifolia are genetically more similar. The populations of Maytenus ilicifolia analyzed formed three groups with low confidence levels. Group I included the populations of Ponta Porã, group II the populations of Santana do Livramento, Vale Verde, Canguçu and Unistalda, and group III was formed by the other 13 populations. A similarity in the genetic and environmental grouping of such species was observed. The chemical characterization was performed by gas chromatography of the extracts obtained with high pressure CO2. An increase in the yield of extract was obtained when the temperature of extraction as well as the solvent density were increased. The compounds that presented higher yield in the first fractions were dodecanoic acid, geranyl acetone, phytol, squalene, vitamin E and stigmast-5-enol. The compounds friedelan-3-ol and friedelin were extracted in higher yields in the following fractions. The chemical variability study of the volatile and semi-volatile compounds of populations of M. ilicifolia showed a yield of extract (0,488 a 0,976 %) significantly different between the populations. In addition no relation between the chemical and environmental or between the chemical and genetic grouping could be established. / Maytenus ilicifolia (espinheira-santa) é uma planta nativa da região sul do Brasil empregada na medicina popular para tratamentos de indigestão, úlceras e gastrites. A ampla utilização de Maytenus ilicifolia na medicina popular, tem levado à perda de populações da espécie, encontrando-se atualmente na lista da FAO como uma das espécies prioritárias para estudo e conservação na América do Sul. Neste sentido, o presente trabalho teve como objetivo realizar estudos que subsidiem protejos de conservação desta espécie. Inicialmente foi realizado um levantamento em herbários e a campo, verificando a ocorrência e distribuição do gênero Maytenus no estado do Rio Grande do Sul, sendo encontradas três novas espécies: M. glaucescens Reiss, M. gonoclada Mart. e M. robusta Reiss. No estudo da variabilidade genética em três populações, a variação intrapopulacional (0,102 a 0,140) é superior à interpopulacional (0,076 a 0,099). As três populações analisadas se agruparam com baixos valores de significância e 7,6% dos alelos presentes são raros. No estudo da variabilidade genética envolvendo 18 populações nativas de Maytenus ilicifolia e plantas de Maytenus aquifolia e Maytenus evonymoidis, pode-se separar as espécies com 100% de confiança e M. ilicifolia e M. aquifolia estão mais próximas geneticamente em relação a M. evonymoidis. As populações de Maytenus ilicifolia analisadas formaram 3 grupos com valores baixos de índice de confiança baixos, sendo o grupo I a população de Ponta Porá, o grupo II as populações de Santana do Livramento, Vale Verde, Canguçu e Unistalda e o grupo III formados pelas 13 populações restantes. Observou-se semelhança no agrupamento genético e ambiental destas populações. No estudo da caracterização química, via cromatografia gasosa dos extratos obtidos por CO2 a alta pressão, observou-se um acréscimo na eficiência de extração com aumento de temperatura e densidade do solvente. Os compostos ácido dodecanóico, geranil acetona, fitol, Esqualeno, Vitamina E e Stigmast-5-enol foram extraídos em maior concentração nas primeiras frações, independente das condições de extração, enquanto que os compostos Friedelan-3-ol e Friedelin, foram extraídos em maior quantidade nas frações seguintes. Na análise da variabilidade química dos compostos voláteis e semi-voláteis das populações de Maytenus ilicifolia ). Verificou-se no rendimento de extrato (0,488 a 0,976 %) e concentração dos compostos analisados diferenças significativas entre as populações analisadas e não houve relação entre o agrupamento químico das populações com os agrupamentos ambientais e genéticos.

Dentro do gênero Adesmia, as técnicas moleculares ainda não foram empregadas na caracterização de germoplasma e na análise da diversidade genética das espécies brasileiras que compôem o gênero. Portanto os objetivos deste trabalho foram: caracterizar, com a utilização de marcador molecular do tipo RAPD, as espécies brasileiras do gênero Adesmia DC; com base nestas informações estabelecer relações de diversidade genética entre as espécies e os acessos analisados; relacionar dados de diversidade com dados morfológicos e de reprodução.

Made available in DSpace on 2014-06-12T15:04:52Z (GMT). No. of bitstreams: 2 arquivo4510_1.pdf: 1312401 bytes, checksum: 103b70bf03d8851fef2766e8bd0290f9 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2004 / Paracoccidioides brasiliensis é um fungo dimórfico e agente etiológico da paracoccidioidomicose, uma micose sistêmica de evolução aguda ou crônica que se não diagnosticada e tratada a tempo pode ser fatal. Um método molecular para caracterização e detecção de P. brasiliensis foi desenvolvido a partir da clonagem e do sequenciamento de um fragmento de DNA de ~750 pb, obtido por RAPD (Random Amplified Polymorphic DNA), presente em isolados virulentos e ausente em isolados avirulentos deste fungo. Uma região interna do fragmento de DNA seqüenciado foi usada para desenhar primers que posteriormente foram utilizados em uma reação de hemi-nested PCR em tubo único. A reação de PCR específica foi capaz de amplificar DNA de três isolados de P. brasiliensis reconhecidamente virulentos e três isolados recentemente obtidos de pacientes com paracoccidioidomicose. A especificidade desta PCR foi confirmada pela ausência de produtos amplificados com DNA genômico de isolados de Histoplasma capsulatum, Blastomyces dermatitidis, Coccidioides immitis, Sporothrix schenckii, Cryptococcus neoformans, Candida albicans, Aspergillus fumigatus, Mycobacterium tuberculosis, Leishmania braziliensis, Trypanosoma cruzi, Schistosoma mansoni, DNA genômico humano (leucócitos) e de isolados de P. brasiliensis reconhecidamente avirulentas. A amplificação de cDNA de um isolado virulento sugere tratar-se de um gene expresso. A detecção específica de isolados virulentos de P. brasiliensis sugere ser este um candidato a marcador de virulência para este fungo. O potencial diagnóstico da PCR específica foi verificado com DNA extraído de aspirado de linfonodo de um paciente com paracoccidioidomicose

Polymorphic light eruption (PLE) is the most common photodermatosis, affecting ~17% of the population. PLE is a delayed-type hypersensitivity response to an antigen induced by solar ultra-violet radiation (UVR). Its effects vary between patients, but the main symptom is a non-scarring, red papular rash in areas exposed to UVR. An effective therapy is low dose ultra-violet B (NBUVB) phototherapy. It is thought that NBUVB phototherapy desensitises the skin to further UVR exposure, but the mechanism by which this happens is unknown. Current immune based studies have been unable to clarify a mechanism as to how PLE arises. However, research in other skin diseases, such as psoriasis and atopic dermatitis, has shown that the barrier function of the skin is compromised by these disorders. Furthermore, research in lesional PLE skin showed an increase in barrier permeability of the skin. Recent research has specifically linked claudin proteins of tight junctions to the barrier dysfunction. Therefore, this study used quantitative immunofluorescent staining to measure tight junction (TJ) proteins and other barrier proteins of interest. Barrier function was also measured by transepidermal water loss (TEWL); a tracer dye penetration assay was used to measure TJ barrier function specifically. All measurements were made in non-lesional PLE skin, as compared to skin from healthy human volunteers. In photoprotected PLE skin the TJ protein claudin-1 was significantly reduced compared to healthy skin. The use of a tracer dye highlighted there was a reduction in TJ barrier function in PLE skin compared to healthy individuals. PLE and healthy skin were then exposed to ultra-violet B (UVB) and 24h later TJ proteins and TJ barrier function were measured. There was no change to claudin-1 after UVB exposure in PLE skin, but claudin-7 was reduced and claudin-12 increased. In contrast, in UVB-irradiated skin in normal controls after UVB exposure claudin-7 and claudin-12 were both increased, whilst claudin-1 was reduced. In PLE patients there was no further change to TJ barrier function, however, in normal controls, skin TJ barrier function was reduced post UVB. Both in healthy and PLE skin TEWL was unchanged before and after UVB exposure. Lastly TJ proteins were investigated after NBUVB in PLE patients. There was a further reduction in claudin-1 in PLE patients as well as a reduction in the TJ protein occludin, however the stratum corneum was significantly thickened. It could be suggested that this is a compensatory measure for the reduction seen in TJ barrier proteins, however further studies are needed to understand this. These data show significant differences in the TJ skin barrier in patients with PLE as compared to healthy human volunteers before and after UVB exposure. Furthermore, in PLE skin there is a significant change to the epidermis after NBUVB phototherapy. These data demonstrate that TJ protein expression and function is altered in PLE skin and may contribute to aetiology of the disorder, however the role of TJ barrier in aetiology is yet to be firmly established.

Polymorphic ventricular tachycardia (PVT) is a life-threatening arrhythmia that is typically related to long QT syndrome, organic heart disease, electrolyte abnormalities, cardiotoxic drugs, or adrenergic stimulation. A review of the literature reveals that PVT with normal QT interval and without underlying cause is quite rare. We report a case of idiopathic spontaneous PVT with structurally normal heart and without electrolyte abnormalities, drug reactions, or evidence of catecholamine induced arrhythmia. We also review the literature on the electrocardiographic characteristics and management of idiopathic PVT.

京都大学 / 0048 / 新制・論文博士 / 博士(農学) / 乙第12834号 / 論農博第2798号 / 新制||農||1026(附属図書館) / 学位論文||H26||N4857(農学部図書室) / 31372 / (主査)教授 栗原 達夫, 教授 植田 和光, 教授 平竹 潤 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・論文博士 / 博士(農学) / 乙第12986号 / 論農博第2826号 / 新制||農||1038(附属図書館) / 学位論文||H28||N4961(農学部図書室) / 32456 / 名古屋大学大学院農学研究科生化学制御専攻 / (主査)教授 祝前 博明, 教授 今井 裕, 教授 廣岡 博之 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Thesis advisor: Vassilios Bezzerides / The aim of this study was to utilize a massively parallel reporter assay (MPRA) to identify organ-specific microRNA (miRNA) target sequences to refine the timing and expression of transgene expression for gene therapy. We previously had developed a cardiac gene therapy for Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT) using a systemically delivered adeno-associated virus (AAV9) vector. We hypothesized that incorporation of organ specific miRNA target sites into our vector construct could improve our therapy’s tissue specificity due to the ability of miRNAs to silence transgene expression. Initially, we attempted to incorporate mir-124 target sequences into our vector to detarget the brain. Although these initial attempts were unsuccessful, the study allowed us to develop a protocol to test the effectiveness of miRNA target sequences. Thereafter, we developed a method to screen thousands of putative miRNA target sequences simultaneously. In this study, target sequences of miRNAs specific to the heart, brain and liver were incorporated into a plasmid library. This plasmid library was subsequently made into AAV and injected into mice from a CPVT transgenic line. Total DNA and RNA was later extracted from the target organs, converted into genomic DNA (gDNA) and complementary DNA (cDNA) libraries respectively, and sent for amplicon sequencing. We analyzed the results using Comparative Microbiome Analysis 2.0 software (CoMA) and a custom python script to count the occurrence of each specified barcode per sample. In doing so, we showed that the miRNA suppression mechanism is not only effective but also organ specific. Furthermore, we developed a second script to create a combinatorial library from a set list of miRNA target sequences enabling us to efficiently test thousands of target sequence combinations at once. In doing so, we will be able to identify effective miRNA target sequence combinations to further improve gene therapy specificity. / Thesis (BS) — Boston College, 2021. / Submitted to: Boston College. College of Arts and Sciences. / Discipline: Departmental Honors. / Discipline: Biology.

In this thesis, a framework for generic uncertainty analysis is developed. The two basic uncertainty characteristics aleatoric and epistemic uncertainty are differentiated. Polymorphic uncertainty as the combination of these two characteristics is discussed. The main focus is on epistemic uncertainty, with fuzziness as an uncertainty model. Properties and classes of fuzzy quantities are discussed. Some information reduction measures to reduce a fuzzy quantity to a characteristic value, are briefly debated. Analysis approaches for aleatoric, epistemic and polymorphic uncertainty are discussed. For fuzzy analysis α-level-based and α-level-free methods are described. As a hybridization of both methods, non-flat α-level-optimization is proposed. For numerical uncertainty analysis, the framework PUQpy, which stands for “Polymorphic Uncertainty Quantification in Python” is introduced. The conception, structure, data structure, modules and design principles of PUQpy are documented. Sequential Weighted Sampling (SWS) is presented as an optimization algorithm for general purpose optimization, as well as for fuzzy analysis. Slice Sampling as a component of SWS is shown. Routines to update Pareto-fronts, which are required for optimization are benchmarked. Finally, PUQpy is used to analyze example problems as a proof of concept. In those problems analytical functions with uncertain parameters, characterized by fuzzy and polymorphic uncertainty, are examined.