Tesi sul tema "Crinane"

Le travail de cette thèse consiste en la synthèse et la désymétrisation d'arylcylohexa-2,5-diène, méthodologie pouvant donner accès à différentes classes d'alcaloïdes. Ces arylcyclohexa-2,5-diènes sont obtenus à partir de biaryles par réduction de Birch alkylante. Dans un premier temps, une étude systématique de cette réaction a été effectuée sur des biaryles substitués par des groupements électrodonneurs. Dans un second temps, les systèmes diéniques ou bis ether d'énols obtenus par cette voie ont été fonctionnalisés par différents processus : dihydroxyloation, iodocyclisation, amination oxydante, et en particulier par hydroamination, réaction réalisée de manière diastéréocontrôlée et menant au squelette hydrocarboné des crinanes. Une réaction de type Mannich a aussi été exploitée pour mener à un système tricyclique, précurseur possible des morphinanes.

Dans le cadre de cette thèse, nous nous sommes intéressés à la synthèse totale d'alcaloïdes de type crinine, de la famille des Amaryllideceae. Notre stratégie de synthèse repose la formation d'un cycle à sept chaînons, avec création concomitante du carbone quaternaire caractéristique de cette famille, par réaction de Heck intramoléculaire. Ainsi, dans un premier temps, les synthèses de l'oxocrinine et de l'oxomaritidine ont été effectuées en sept étapes avec, respectivement, 22. 1% et 14. 8% de rendement. Dans un deuxième temps, la crinine, la maritidine et la buphanisine, éther méthylique de la crinine, ont été formées, en deux étapes, par réduction diastéréosélective d'énone suivie de l'inversion de configuration de l'alcool par SN2 ou par réaction de Mitsunobu. Les synthèses de la flexinine et de l'augustine, alcaloïdes possédant une fonction époxyde, ont été envisagées de multiples façons pour conduire finalement à l'époxy-alcool avec la stéréochimie adéquate. Malheureusement, les premiers essais d'inversion de configuration de l'alcool ont échoué et, n'ont donc pas permis d'accéder aux molécules naturelles cibles. Enfin, la synthèse de ces alcaloïdes a permis, par réarrangement de type diénone-phénol, de former à partir des intermédiaires spirodiénones, des dérivés biaryliques pontés, analogues de la buflavine, alcaloïdes des Amaryllidaceae
Within the framework of this thesis, we were interested in the total synthesis of crinine-type alkaloids, belonging to Amaryllideceae plants. Our strategy is based on the formation of seven membered ring and on the simultanous creation of critical quaternary carbon by intramolecular Heck reaction. First, oxocrinine and oxomaritidine have been synthetised in seven steps in, respectively, 22. 1% and 14. 8% overallyield. Secondly, maritidine, crinine, its methyl ether, the buphanisine have been obtained, in two steps, by diastereoselective enone reduction, followed by SN2 alcohol inversion or by Mitsunobu reaction. The flexinine and augustine syntheses were considered in different ways to get, finally, the epoxy-alcohol with the correct stereochemistry. Unfortunately, the first attempts of alcohol inversion failed, our synthetical target was not obtained. Finally, dienone-phenol rearrangement of the synthetised spirodienone intermediates led to the bridged biaryl compounds, buflavine analogs

Thesis (M.S.)--University of Maryland, College Park, 2005.
Thesis research directed by: Dept. of Chemistry and Biochemistry. Title from t.p. of PDF. Includes bibliographical references. Published by UMI Dissertation Services, Ann Arbor, Mich. Also available in paper.

Il presente elaborato si propone di analizzare il racconto dalla scrittrice argentina María Teresa Andruetto (Premio Hans Christian Andersen 2012) El país de Juan, nell’edizione pubblicata dalla casa editrice spagnola Anaya nel 2003. Nel 2014 ne è stata proposta la traduzione in italiano con il titolo Il paese di Juan, ad opera di Ilide Carmignani. La storia, che narra di due bambini costretti a migrare dalla campagna alle favelas argentine (Villas), si presenta come una vera e propria sfida traduttiva per il contrasto tra la poeticità del linguaggio e il realismo del mondo raccontato ed il forte legame con il contesto locale in cui è inserita. Lo studio consta di tre capitoli principali: nel primo si approfondiscono la vita, le opere, lo stile e la concezione della letteratura della Andruetto. Dopo aver impostato tale cornice stilistica e contestuale, si prosegue, nel secondo capitolo, con l’analisi del testo originale, dove vengono messi in luce gli aspetti formali e stilistici della scrittura, le tematiche, gli elementi grafici, le illustrazioni e il contesto storico-sociale. Si passa poi, nel terzo ed ultimo capitolo, ad un’analisi della traduzione italiana del libro rispetto alla capacità di ricreare un tipo di linguaggio altamente innovativo, la salvaguardia delle figure retoriche, degli aspetti grafici e dei riferimenti culturali, elementi di fondamentale importanza per creare una traduzione in grado di aprire gli orizzonti del giovane lettore. Inoltre, si confrontano i glossari della versioni spagnola e italiana evidenziandone differenze e affinità. Si pone infine l’accento sull’indiscutibile abilità della Carmignani nel saper mantenere la dicotomia testuale tra realismo e mistero, riferendosi anche all’importanza della collaborazione tra lo scrittore e il traduttore, legame che senza dubbio è possibile cogliere nella traduzione Il paese di Juan.

Os efeitos individuais e interativos dos parâmetros ambientais físicos e químicos, como temperatura, intensidade luminosa, salinidade e concentração de fósforo inorgânico dissolvido na água do mar, na produção de proteínas, carboidratos, acúmulo de fósforo tecidual e taxa de absorção do fósforo inorgânico disponível no meio de cultura em Gelidium crinale (Turner) Lamouroux, foram investigados durante um período de sete dias de cultivo laboratorial, em condições controladas. A ação dos parâmetros abióticos foi analisada de três maneiras diferentes. A primeira avaliação integrou a ação de temperatura, intensidade luminosa e fósforo inorgânico dissolvido, mantendo-se fixa a salinidade em 25 ups, onde se constatou que em todos os componentes químicos algais ocorreram interações de terceira ordem. O incremento de 2,28 a 2,67 % nos teores de proteínas foram obtidos à temperatura de 25 °C e 12 μmol m-2 s-1 de intensidade luminosa, diminuindo com a elevação da intensidade luminosa para 40 μmol m-2 s-1. Para carboidratos, ocorreram interações significativas entre os três parâmetros, com um aumento de 6,85 % sendo registrado a 25 °C de temperatura, 24 μmol m-2 s-1 de intensidade luminosa e 10,0 μM de fósforo inorgânico. O aumento máximo na taxa de fósforo tecidual (0,56 %) ocorreu em talos cultivados nas menores temperatura e intensidade luminosa e na maior concentração de fósforo inorgânico dissolvido. Com relação à intensidade luminosa, foi observada uma correlação negativa entre proteínas e carboidratos. A segunda avaliação estabeleceu a ação independente e sinérgica de temperatura, salinidade e fósforo inorgânico disponível no meio de cultivo, fixando-se a intensidade luminosa em 24 μmol m-2s-1. A maior produção de proteínas ocorreu em cultivos onde a temperatura foi de 25 °C, com uma concentração de 5,0 e 10,0 μM de fósforo inorgânico dissolvido e salinidade entre 15 e 20 ups, cujos valores médios do incremento variaram entre 2,62 a 2,83 % peso seco de alga, resultando em uma interação de terceira ordem altamente significativa. Para carboidratos a elevação de 6,85 % em sua concentração está associada à maior temperatura (25 °C), maior salinidade (25 ups) e maior quantidade de fósforo inorgânico disponível no meio de cultivo (10,0 μM). Contudo, não foi observada uma interação de terceira ordem através da análise estatística. Para esta biomolécula observaram-se interações de segunda ordem altamente significativa (P < 0,005) entre temperatura e diferentes concentrações de fósforo inorgânico e entre temperatura e salinidade (P < 0,000). O acúmulo de fósforo nos talos da alga foi menor durante os cultivos em que a salinidade foi de 25 ups,nas temperaturas de 20 e 25 °C e concentração de fósforo disponível de 2,5 μM, com percentuais entre 0,08 a 0,11 % em peso de cinzas. O maior incremento ocorreu na menor temperatura, associada à baixa salinidade e alta concentração de fósforo inorgânico no meio. O coeficiente de correlação de Pearson revelou correlações positivas, altamente significativas (P < 0,001) entre teor de proteína, temperatura e disponibilidade de fósforo inorgânico no meio de cultivo. Para carboidratos, as correlações foram positivas com os três parâmetros abióticos. Para fósforo tecidual somente com o fósforo inorgânico disponível no cultivo foi que ocorreu uma relação positiva; com os outros dois parâmetros esta correlação foi negativa. Entre os componentes químicos encontrados nas algas, proteínas e carboidratos apresentaram uma relação positiva, porém fósforo tecidual apresentou uma correlação negativa com ambos, embora com proteínas esta relação não tenha sido significativa. A terceira avaliação estudou a ação individual e o sinergismo entre os parâmetros ambientais, temperatura, intensidade luminosa e salinidade, a uma concentração fixa de fósforo inorgânico disponível no meio de cultivo (10,0 μM), sobre a composição química, bem como na taxa de absorção de fósforo inorgânico disponível. Observou-se a ocorrência de interações de terceira ordem em todos as variáveis estudadas. O teor de proteínas apresentou um aumento de 3,72 % durante o período de cultivo, passando de 20,63 % antes do cultivo, para 24,35 % após o término do experimento, principalmente nas condições de 25 °C de temperatura, 12 μmol m-2s-1 de intensidade luminosa e 15 ups de salinidade. Para carboidratos, nas condições de baixa intensidade luminosa (12 μmol m-2s- 1), a uma temperatura de 20 °C e salinidades de 10 e 15 ups, foram registrados valores inferiores à amostra controle, caracterizando um consumo desta biomolécula por parte das algas. Nestas mesmas condições ambientais, foram registrados os maiores teores de fósforo tecidual, variando entre 0,86 a 1,09 % do peso das cinzas. As maiores taxas de absorção do fósforo do meio ocorreram na salinidade de 25 ups e 25 °C de temperatura, diminuindo da intensidade luminosa de 12 μmol m-2s-1 para 40 μmol m-2s-1. As maiores concentrações de fósforo inorgânico residual na água do meio de cultivo ocorreram nas salinidades de 10 e 15 ups, em todas as intensidade luminosas e temperaturas estudadas. Através do coeficiente de correlação de Pearson, observou-se que os teores de proteínas apresentaram uma forte correlação negativa com a intensidade luminosa e positiva com a temperatura e salinidade, embora com esta última não tenha sido significativa. Para carboidratos, as correlações com os parâmetros abióticos foram todas positivas. Correlações negativa e positiva, não significativas, foram observadas entre esta biomolécula e o teor de proteínas e a taxa de absorção de fósforo disponível no meio, respectivamente. Por outro lado, com fósforo tecidual, ocorreu uma correlação negativa, altamente significativa. Este estudo mostra o estado fisiológico de Gelidium crinale e contribui para o estabelecimento das melhores condições de cultivo para produção de proteína, carboidrato e fósforo tecidual e indicação do uso racional de nutrientes, fornecendo informações para a otimização de processos de maricultura, tanto em termos de cultivo bem sucedido de algas, quanto de redução no impacto sobre o ambiente.
The individual and interactive effects of the physical and chemical environmental parameters such as temperature, light intensity, salinity and dissolved inorganic phosphorus concentration in the sea water, on protein and carbohydrate production, tissue phosphorus accumulation and absorption rate of inorganic phosphorus available in the culture medium by Gelidium crinale, were investigated in laboratory cultures, for seven days, under controlled conditions. The action of the abiotic parameters was analyzed in three different ways. The first evaluation integrated the action of temperature, light intensity and dissolved inorganic phosphorus, at a constant salinity of 25 ups, where third-order interactions were verified for all algal chemical compounds. The increase of 2,28 to 2,76 % in the protein content was obtained at a temperature of 25 °C and 12 μmol m-2 s-1 of light intensity, decreasing with an increase in light intensity towards 40 μmol m-2 s-1. For carbohydrates there was a significant interaction among the abiotic parameters, with a maximum increment of 6.85 % at 25 °C of temperature, 24 μmol m-2 s-1 of light intensity and 10.0 μM of inorganic phosphorus. The highest increase on tissue phosphorus concentration (0.56 %) occurred in thalli grown under the lowest temperature and light intensity and the highest dissolved inorganic phosphorus concentration used in this study. With regard to light intensity, was observed a negative correlation between proteins and carbohydrates. The second evaluation established the independent and synergic action of temperature, salinity and available inorganic phosphorus, setting the light intensity at 24 μmolm-2s-1. The maximum production of proteins occurred in cultures where the temperature was 25 °C, with a concentration of 5,0 and 10,0 μM of dissolved inorganic phosphorus and salinity between 15 and 20 ups, with values varying among 2,62 to 2,83% of algae dry weight, resulting in a highly third order significant interaction. For carbohydrates the elevation of 6,85% in concentration is associated to the greatest temperature (25 °C), salinity (25 psu) and amount of available inorganic phosphorus in the culture (10,0μM). However, a third-order interaction was not observed in the statistical analysis. For this biochemical compound, only a second order interaction was observed between temperature and inorganic phosphorus concentrations (P < 0,005) and between temperature and salinity (P < 0,000). The phosphorus accumulation in the thalli was minor during growth under salinity of 25 ups, temperatures of 20 and 25 °C and dissolved inorganic phosphorus concentration of 2,5 μM, with values of 0,08 and 0,11% on an ash weight basis. The greatest increase occurred in the smallest temperature, associated to low salinity and high inorganic phosphorus concentration in the culture. The Pearson’s correlation coefficient revealed highly significant (P < 0,001) positive correlations among protein content, temperature and inorganic phosphorus availability in the growth medium. For carbohydrates, correlations were positive with all three abiotic parameters. For tissue phosphorus, a positive correlation occurred only with dissolved inorganic phosphorus; with temperature and salinity the correlations were negative. Among the chemical components present in the algae, proteins and carbohydrates showed a positive relation, while tissue phosphorus presented a negative correlation with both, although this correlation was not significant with regard to protein. The third evaluation looked at the individual and synergic action among the environmental parameters of temperature, light intensity and salinity, at a constant concentration of dissolved inorganic phosphorus (10,0 μM) on the chemical composition, as well as in absorption rates of available inorganic phosphorus. Third-order interactions were observed for all the studied variables. Proteins presented an increase of 3,72% during the growth period, ranging from 20,63 %, before cultivation to 24,35% by the end of the experiment, especially at 25 °C of temperature, 12 μmolm-2s-1 of light intensity and salinity of 15 ups. For carbohydrates, under conditions of low light intensity (12 μmolm-2s- 1), a temperature of 20 °C and salinities of 10 and 15 ups, concentrations were below the control values, characterizing a consumption of this component by the algae. In these same environmental conditions, the largest quantity of tissue phosphorus was registered, varying from 0,86 to 1,09% of ash weight. The highest absorption rates of the phosphorus available in the medium took place under salinity of 25 ups and 25 °C of temperature, decreasing from a light intensity of 12 μmolm-2s-1 to 40 μmolm-2s-1. The highest concentrations of residual inorganic phosphorus in the medium seawater were measured for salinities of 10 and 15 ups, for all the light intensities and temperatures studied. Analysis of the Pearson’s correlation coefficient demonstrated that proteins content had a strong negative correlation with light intensity and positive correlation with temperature and salinity, although with this last one the correlation was not significant. For carbohydrates, the correlations with the abiotic parameters were all positive. Negative and positive correlations, though not significant, were observed among this biochemical component and protein content and absorption rate of available inorganic phosphorus, respectively. On the other hand, with tissue phosphorus, carbohydrates showed a highly significant negative correlation. This study reveals the physiological behavior of Gelidium crinale, and contributes to the establishment of the optimum conditions of the growth medium and to the rational use of nutrients, providing valuable information for optimizing processes of marine culture, both in terms of successful growth of algae and reduced impact on the environment.

Thesis (Ph. D.) -- University of Maryland, College Park, 2006.
Thesis research directed by: Cell Biology & Molecular Genetics. Title from t.p. of PDF. Includes bibliographical references. Published by UMI Dissertation Services, Ann Arbor, Mich. Also available in paper.

Thesis (Ph. D.) -- University of Maryland, College Park, 2006.
Thesis research directed by: Cell Biology & Molecular Genetics. Title from t.p. of PDF. Includes bibliographical references. Published by UMI Dissertation Services, Ann Arbor, Mich. Also available in paper.

Infection of the resistant Arabidopsis ecotype Di-17 with Turnip Crinkle Virus (TCV) elicits hypersensitive response (HR), accompanied by increased expression of defense genes. HR to TCV is conferred by HRT, which encodes a coiled-coil (CC)-nucleotide-binding site (NBS)-leucine-rich repeat (LRR) class of resistance (R) protein. In contrast to HR, resistance requires HRT and a recessive locus designated rrt. Unlike most CC-NBS-LRR R proteins, HRT-mediated resistance is dependent on EDS1 and independent of NDR1. Resistance is also dependent on salicylic acid (SA) pathway and light. A dark treatment, immediately following TCV inoculation, suppresses HR, resistance and activation of a majority of the TCV-induced genes. To determine the genetic, molecular and biochemical basis of light-dependent defense pathway, we studied the role of various photoreceptors in HRT-mediated resistance to TCV, HRT protein levels and its localization. Interestingly, mutation in blue-light photoreceptors led to degradation of HRT via a proteasome-dependent pathway and resulted in susceptibility to TCV. Exogenous application of SA induced transcription of HRT, which restored HRT levels in some, but not all, mutant backgrounds. These results show that different photoreceptors function distinctly in maintaining post-transcriptional stability of HRT. In addition to photoreceptors, HRT also forms a complex with several other proteins, many of which participate in the RNA silencing pathway and are required for HRT-mediated resistance. Together, our results suggest that HRT forms a multi-protein complex and that HRT-mediated signaling involves reconstitution of this complex.

Thesis (M.S.)--Worcester Polytechnic Institute.
Keywords: Turnip crinkle virus; arabidopsis; thaliana; TCV; avrRpt2; avrRpm1; avrRps4; systemic acquired resistance; virulence; Avr gene; R gene; pseudomonas syringae. Includes bibliographical references (p. 60-66).

La Lipoproteïna (a) és una classe de lipoproteïna semblant a les lipoproteïnes de baixa densitat (LDL), en la que l'apo B100 està unida de forma covalent a una glicoproteïna molt polimòrfica de mida, anomenada apo(a).
Si bé la seva funció fisiològica és desconeguda, nombrosos estudis epidemiològics han demostrat que la concentració plasmàtica elevada de Lp(a) representa un factor de risc independent de desenvolupar una malaltia cardiovascular. Alguns d'aquests estudis però han donat lloc a resultants contradictoris.
L'apo(a) comparteix amb el plasminogen una forta homologia estructural. Com el plasminogen, s'estructura en dominis proteics o Kringles que contenen regions anomenades LBS (Lysine Binding Site)capaces d'interaccionar amb els residus de lisina de les proteïnes. En l'apo(a) el Kringle IV10 confereix la funció LBS a la Lp(a).

El propòsit d'aquest estudi ha estat el de:

· Examinar l'associació entre la concentració de Lp(a), la mida de l'apo(a) i la funció LBS en pacients amb infart agut de miocardi prematur i els seus respectius controls.
· Cercar mutacions en kringle IV10 de l'apo(a) que puguin alterar l'activitat LBS de la Lp(a).
· Investigar l'efecte que l'emmagatzematge de les mostres congelades durant 5 anys té en la concentració de Lp(a) i analitzar la relació d'aquest efecte amb el número de repeticions del kringle IV dels seus al·lels.

Mètodes

La concentració plasmàtica de Lp(a) va ser mesurada per un mètode immunoturbidimètric, el genotip d'apo(a) es va determinar amb una modificació de la tècnica de pulsed-field gel electrophoresis descrita per Lackner i col., el fenotip d'apo(a) amb una electroforesi en gel d'agarosa i immunoblotting, l'activitat LBS es va quantificar amb un immunoassaig descrit per Hoover-Plow i col. i la detecció de mutacions amplificant la regió del kringle IV10 i aplicant al producte de PCR la tècnica de SSCP .

Resultats

Vegeu :

Simo JM, Camps J, Vilella E, Gomez F, Paul A, Joven J. Instability of lipoprotein(a) in plasma stored at -70 degrees C: effects of concentration, apolipoprotein(a) genotype, and donor cardiovascular disease. Clin Chem. 2001 Sep;47(9):1673-8.

Simo JM, Joven J, Vilella E, Ribas M, Figuera L, Virgos C, Sundaram IM, Hoover-Plow J. Polymorphisms in human apolipoprotein(a) kringle IV-10 and coronary artery disease: relationship to allele size, plasma lipoprotein(a) concentration, and lysine binding site activity. J Mol Med. 2001 Jun;79(5-6):294-9.

Simo JM, Joven J, Vilella E, Ribas M, Pujana MA, Sundaram IM, Hammel JP, Hoover-Plow JL. Impact of apolipoprotein(a) isoform size heterogeneity on the lysine binding function of lipoprotein(a) in early onset coronary artery disease. Thromb Haemost. 2001 Mar;85(3):412-7.


· En els pacients la concentració de Lp(a) es més elevada, la mida de l'apo(a) més petita i l'activitat LBS més alta en les isoformes més petites comparat amb els controls.

· Hem identificat una nova mutació en heterozigosi en un pacient. La mutació W81R identificada prèviament en humans no ha estat trobada en la nostra mostra en canvi la mutació M75T és freqüent. Aquesta mutació no modifica la funció LBS però el genotip TT s'associa amb risc de patir una malaltia cardiovascular.

· Durant l'emmagatzematge de les mostres la Lp(a) perd immunoreactivitat de forma significativa en pacients però no en controls. Aquesta pèrdua no es relaciona amb el número de kringle i es limita a mostres amb concentracions inicials entre 41 i 345 mg/L.

Conclusions

· Aquest estudi suggereix una associació entre malaltia cardiovascular, activitat LBS elevada i isoformes petites.

· La mutació W81R, si està present, és molt poc freqüent en la població espanyola.

· La concentració plasmàtica de Lp(a) dels pacients és menys estable que la dels controls, les diferències no es relacionen amb la distribució dels genotips però poden ser dependents de la concentració Aquestes diferències poden complicar la interpretació d'algun estudis.
Lipoprotein (a) is a class of lipoprotein particles resembling low density lipoprotein (LDL) in which apo B100 is covalently linked to a high polymorphic glycoprotein apolipoprotein termed apo(a).
Although the physiological function of Lp(a) is unknown, numerous epidemiological studies have shown that a high plasma concentration of Lp(a) represent an independent risk factor for the development of coronary heart disease, however some confusing results in some of this studies has been reported.
Apo(a) shares a high structural homology with plasminogen As plasminogen, apo(a) kringle domains contain Lysine Binding Site(s) (LBS) that interact with residues of lysine. Kringle IV10 of apo(a) is the primary LBS of Lp(a) and is associated with lesion formation on transgenic mice .

The purpose of this study was

· To examine de association of Lp(a) concentration, apo(a) size and Lp(a) lysine-binding site in patients with early onset heart diseases and age matched controls.
· Search for mutations in the apo(a) kringle IV10 which could alter the LBS activity of Lp(a)
· to investigate the decrease in Lp(a) values in samples from controls and patients with established cardiovascular disease that had been frozen for 5 years and to analyze the relationship between such decrease and the number of kringle IV repeats in the smallest and largest isoforms.

Methods

Blood was obtained from survivors of premature myocardial infarction and age matched controls, plasma was measured by immunoturbidimetry , apo(a) genotype was determined by pulsed-field gel electrophoresis, apo(a) phenotype by gel electrophoresis and immunoblotting, LBS activity was measured by a quantitative LBS-Lp(a) immunoassay as previously described. Detection of mutations and polymorphisms in kringle IV10 was revealed by PCR amplification and applying SSCP analysis to the product

Results
See also:

Simo JM, Camps J, Vilella E, Gomez F, Paul A, Joven J. Instability of lipoprotein(a) in plasma stored at -70 degrees C: effects of concentration, apolipoprotein(a) genotype, and donor cardiovascular disease. Clin Chem. 2001 Sep;47(9):1673-8.

Simo JM, Joven J, Vilella E, Ribas M, Figuera L, Virgos C, Sundaram IM, Hoover-Plow J. Polymorphisms in human apolipoprotein(a) kringle IV-10 and coronary artery disease: relationship to allele size, plasma lipoprotein(a) concentration, and lysine binding site activity. J Mol Med. 2001 Jun;79(5-6):294-9.

Simo JM, Joven J, Vilella E, Ribas M, Pujana MA, Sundaram IM, Hammel JP, Hoover-Plow JL. Impact of apolipoprotein(a) isoform size heterogeneity on the lysine binding function of lipoprotein(a) in early onset coronary artery disease. Thromb Haemost. 2001 Mar;85(3):412-7.


· In the patients, Lp(a) concentration was higher, apo(a) size was smaller, and LBS activity higher in the small isoforms compared to the controls.

· We identified a novel mutation in a patient heterozygous for the mutation The mutation W81R previously identified in humans was not found in our sample, but the M75T mutation was frequent.. The genotype TT conferred a significant risk for myocardial infarction but was not associated with the LBS function of apo(a).

· During storage, mean Lp(a) decreased significantly in samples from patients but not in samples from controls This was not related to the kringle number and was limited to samples with initial plasma Lp(a) concentrations between 41 and 345 mg/L.

Conclusions

· This study suggests an association of high LBS activity in small isoforms of Lp(a) with disease in humans.

· W81R mutation if present is infrequent in Spanish population

· Plasma Lp(a) from patients is less stable than Lp(a) from controls, and the difference is not related to distribution of apo(a) genotypes but may be concentration-dependent. Differential sample stability may complicate the interpretation of several studies.

La socavación es una de las operaciones críticas dentro del proceso productivo en una mina explotada por hundimiento de bloques o paneles. El conocimiento del proceso de socavación proviene en gran medida de la experiencia operativa, sin un mayor conocimiento teórico. Por lo tanto, resulta fundamental mejorar el entendimiento de este proceso, y apuntar hacia la confección de guías de diseño ingenieril. La revisión del estado del arte a la fecha mostró que la tendencia actual en el diseño se orienta a la selección del hundimiento avanzado como estrategia de socavación, en general optando por geometrías de corte basal alto, en particular corte inclinado o hundimiento frontal. En el presente trabajo se muestra una comparación geotécnica relativa entre las técnicas de socavación que fueron consideradas con mayor potencial de aplicación dentro de las identificadas en la revisión de diseños: por un lado, hundimiento convencional, y por otro, las variantes de hundimiento avanzado, corte plano e inclinado. Modelación numérica elástica mediante el software Map3d fue realizada para entender el impacto de las distintas geometrías de corte basal, sobre dos aspectos críticos: los esfuerzos inducidos en la frente de socavación y la condición de los pilares de los niveles de producción y hundimiento. La estrategia de modelación desarrollada fue del tipo paramétrica, modificando una variable de interés a la vez: la secuencia de socavación, la geometría del corte; la razón entre los esfuerzos horizontal y vertical in situ y la profundidad. Para comparar entre las variantes, se utilizaron dos indicadores: la razón entre el esfuerzo vertical inducido, medido en los pilares del nivel de hundimiento posterior a la socavación, y el esfuerzo vertical in situ; y un factor de seguridad, asumiendo el criterio de falla de Mohr-Coulomb, aplicado sobre los pilares de los niveles de producción y hundimiento. Los resultados obtenidos muestran que la estrategia que parece ser más robusta corresponde a la de corte inclinado avanzado, debido a que en la mayoría de los escenarios analizados, ofrece las condiciones de diseño más benignas: mayores factores de seguridad sobre los pilares y bajos esfuerzos inducidos en el nivel de hundimiento, en particular con respecto al caso de corte plano.

Hoy, en el mundo del arte de Lima, se puede percibir un fenómeno social definido por Robert Hughes (1992) como “Cultural Cringe”. Dicho fenómeno se basa en la incapacidad que tiene el artista para fundamentar su obra en una sociedad en la que el consumidor se ve condicionado por una exposición constante a medios de comunicación masivos. Al parecer, la solución se encuentra en la propuesta de una Escuela de Arte en Barranco, escenario principal de dicho “Cringe”. Dicha casa de estudios debe poder ofrecer una educación integral, teórica y práctica, que permita que los alumnos sean competitivos en una realidad internacional y, eventualmente, debe permitir que el contexto social en el que se encuentra se vea contagiado por el conocimiento y la cultura que la misma imparte.
Defined by Robert Hughes (1992);"Cultural Cringe" is a social phenomenon that can be perceived in Lima’s Art world. This phenomenon occurs when artists are unable to demonstrate their work as conceptually and intellectually coherent pieces, especially as a result of constant exposure to “garbage” mass media. Apparently, said “Cringe” can be corrected with an Art School Project in Barranco, where the problem is more latent. The proposed School should be able to offer a comprehensive, theoretical and practical education that allows students to be competitive in an international reality and, eventually, be the kick starter of a new knowledge and culture based set of customs.
Tesis

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The use of animal models in biomedical research is ever increasing. Models that use primates might also have advantages in terms of low maintenance costs and availability of biological knowledge, thereby favoring their use in different experimental protocols. Many current stress studies use animal models at different developmental stages since biological response differs during ontogeny. The aims of this study were to perform a detailed characterization of the developmental stages of common marmosets (Callithrix jacchus), a very important animal model used in biomedical research. Ten subjects, 6 females and 4 males, were followed from birth to initial adult age (16 months). Behavioral and fecal collection for measurement of adrenal (cortisol) and sex (progesterone, estradiol and androgens) hormones took place twice a week during the first month of life and once a week for the remainder of the study. Behavior was observed for 30 minutes in the morning (0700-09:00h) and afternoon (12:00-14:00h). Behavioral profile showed changes during ontogeny, characterizing the 4 developmental stages and the respective phases proposed by Le?o et al (2009).. Differentiation of developmental stages was considered using the onset, end, change and stabilization of the behavioral profile parental care (weaning and carrying), ingestion (solid food), affiliation (social grooming) and autogrooming, agonism (scent marking and piloerection) and play behavior and endocrine profile. Infant weaning and carrying terminated within the infantile stage and the peak of solid food ingestion was recorded in the infantile III phase. Receiving grooming was recorded earlier than grooming performed by the infant and autogrooming. The first episode of scent marking was recorded in the 4th week and it was the least variable behavior, in terms of its onset, which, in almost all animals, was between the 5th and 7th week of life. Solitary play and play with the twin started around the 7th week and play with other members of the group started 8 weeks later. Sex hormone secretion started to differ from basal levels between the 21st and 23rd week of life, in males and females, suggesting that puberty occurs simultaneously in both sexes. Basal cortisol, even at an early age, was higher in females than in males. However, cortisol was not correlated with the juvenile stage, as expected, since this stage corresponds to the transition between infancy and adult age and most behaviors are intensified by this time. The behavioral and endocrine profile of subadult animals did not differ from that of the adults. These results provide more detailed parameters for the developmental process of C. jacchus and open new perspectives for the use of experimental approaches focused on the intermediate ontogenetic phases of this species
O uso de modelos animais em pesquisa biom?dica ? cada vez mais crescente e os modelos utilizando primatas devem apresentar vantagens em termos de custos de manuten??o e caracter?sticas relacionadas ao maior conhecimento da sua biologia para uso em diferentes protocolos experimentais. Diferentes modelos atuais de pesquisa em estresse utilizam animais em diferentes est?gios do desenvolvimento e in?meros estudos demonstram a diferen?a na resposta biol?gica diante de agentes estressores ao longo da ontog?nese. Com o objetivo de caracterizar de maneira mais detalhada as fases do desenvolvimento do sagui comum, Callithrix jacchus, importante modelo utilizado em pesquisa biom?dica, utilizou-se 10 animais, 6 f?meas e 4 machos, que foram acompanhados desde o nascimento at? o in?cio da idade adulta (16 meses). A coleta de dados comportamentais e de fezes para a mensura??o dos horm?nios esteroides de origem adrenal (cortisol) e gonadais (progesterona, estradiol e andr?genos) foram realizadas duas vezes por semana no primeiro m?s de vida dos filhotes e semanalmente no restante do estudo. As observa??es comportamentais tiveram dura??o de 30 minutos e foram realizadas nos turnos matutino (07:00-09:00h) e vespertino (12:00-14:00h).O perfil comportamental de C. jacchus apresentou modifica??es ao longo da ontog?nese caracterizando cada um dos 4 est?gios ontogen?ticos e suas respectivas fases propostas na classifica??o de Le?o (2009). A diferencia??o das etapas do desenvolvimento foi feita a partir do surgimento, t?rmino, varia??o e estabiliza??o no perfil comportamental - cuidado parental (amamenta??o, transporte) ingest?o de alimentos (alimenta??o s?lida), afilia??o (cata??o social recebida e feita e autocata??o) agonismo (marca??o de cheiro e piloere??o) e brincadeira - e no perfil end?crino. O comportamento de amamenta??o e transporte terminou na fase infantil II e a ingest?o alimentar apresentou seu pico na fase infantil III. A cata??o social recebida antecedeu a cata??o feita e a autocata??o, e o comportamento de marca??o de cheiro foi o que apresentou menor variabilidade em rela??o ao seu in?cio, que se concentrou, em quase todos os animais, entre a 5? e 7? semanas de vida. A brincadeira solit?ria e com o g?meo principiaram ao redor da 7? semana e a brincadeira com outros membros do grupo se iniciou 8 semanas depois. A secre??o dos horm?nios sexuais passou a se diferenciar dos valores basais entre a 21? e 23? semanas em machos e f?meas, sugerindo que a puberdade acontece simultaneamente para os dois sexos. O cortisol basal de f?meas mesmo em idade imatura foi mais elevado do que nos machos. Contudo, as varia??es do cortisol n?o se correlacionaram com o est?gio juvenil como esperado uma vez que esse est?gio corresponde ? transi??o entre a idade infantil e a idade adulta, e que se expressou com a intensifica??o da maioria dos comportamentos. Os padr?es comportamentais e end?crinos dos adultos n?o diferiram entre as idades subadulta e adulta. Estes resultados disponibilizam par?metros de desenvolvimento mais detalhados para C. jacchus e abrem perspectivas para a utiliza??o de abordagens experimentais focadas em determinadas etapas da ontog?nese dessa esp?cie

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Objective- Convinced that periodontium, many times, can show alterations in human health, the aim of these studies was to investigate the periodontal situation in patients with endocrine-metabolic disorders such as, Berardinelli-Seip Syndrome, hyperthyroidism, hypothyroidism and acromegaly. Methods- Eight patients with Berardinelli-Seip Syndrome, 16 acromegalics, 30 hypothyroids, 30 hyperthyroids, and a control group with 35 patients were evaluated. Clinical attachment loss, probing depth, gingival bleeding index, gingival overgrowth and Index of Decayed, Missing and Filled Teeth were measured in each patient. All ethical aspects were rigidly observed, being the study conducted after its approval by the University of Fortaleza Research Ethics Committee. Results- The presence of periodontitis was marked in hyperthyroids and in patients with Berardinelli-Seip Syndrome. Hypothyroids showed not much presence of periodontitis, while all acromegalics presented absence of periodontitis. Conclusions- The protective effect of periodontitis in acromegalic patients is a new finding, whose mechanisms are not yet clear, but may be related to the anabolic effects of growth hormone. The presence of periodontitis in Berardinelli-Seip Syndrome may occur due the early onset of diabetes. In hyperthyroids, the high prevalence of periodontitis could be linked to thyroid hormones effects on bone, explaining also the minor prevalence in hypothyroids
Objetivo- Com a convic??o de que o periodonto, muitas vezes, pode ser um sinalizador de desequil?brios da sa?de humana, os presentes estudos tiveram como objetivo investigar a situa??o periodontal de pacientes com dist?rbios end?crino-metab?licos, destacando-se a S?ndrome de Berardinelli-Seip, hipertireoidismo, hipotireoidismo e acromegalia. M?todos- Foram avaliados 8 pacientes com S?ndrome de Berardinelli-Seip, 30 hipertire?ideos, 30 hipotire?ideos e 16 acromeg?licos, al?m de 35 pacientes que fizeram parte do grupo controle. Perda de inser??o cl?nica, profundidade de sondagem, ?ndice de sangramento gengival, aumento gengival e ?ndice de dentes cariados, perdidos e obturados foram os par?metros investigados em cada paciente. Todos os aspectos ?ticos foram rigidamente observados, sendo o estudo conduzido ap?s a aprova??o pelo Comit? de ?tica em Pesquisa da Universidade de Fortaleza. Resultados- A presen?a de periodontite foi marcante nos pacientes hipertire?ideos e naqueles com S?ndrome de Berardinelli-Seip. Os pacientes hipotire?ideos mostrarm pouca presen?a de periodontite, enquanto todos os acromeg?licos apresentaram aus?ncia de periodontite. Conclus?o- O efeito protetor dos acromeg?licos em rela??o ? periodontite ? um novo achado cujos mecanismos ainda n?o est?o claros, mas podem estar relacionados com os efeitos anab?licos do horm?nio de crescimento. A presen?a de periodontite na S?ndrome de Berardinelli-Seip pode ocorrer devido ? precoce presen?a de diabetes. Nos hipertire?ideos, a alta preval?ncia de periodontite pode estar ligada aos efeitos dos horm?nios tireoideanos no osso, explicando a menor preval?ncia nos hipotire?ideos

Replication of genetic material is the most important and central process during the viral life cycle. Most RNA viruses assign one or more proteins translated from their own genome for replicating genomic RNAs. Understanding the various biochemical activities of these replication proteins is the aim of this dissertation research. The replicase proteins of Turnip crinkle virus (TCV) and Tomato bushy stunt virus (TBSV) were selected for this study. Both viruses have small, messenger-sense, single-stranded RNA genomes. Replicase proteins p28/p88 of TCV and p33/p92 of TBSV- were expressed and purified from E. coli as N-terminal fusions to maltose binding protein. In vitro assays revealed that the recombinant p88 has RNA-dependent RNA polymerase (RdRp) and RNAbinding activities. Deletion of the N-terminal p28 domain in p88 resulted in a highly active RdRp, while further deletions at both N- and C-terminal ends abolished RdRp activity. Comparison of p88, the N-terminal p28-deletion mutant of p88 and a TCV RdRp preparation obtained from infected plants revealed remarkable similarities in RNA template recognition and plus and minus strands synthesis. Contrary to recombinant TCV RdRp activities under similar experimental conditions. p33 preferentially binds to singlestranded (ss) RNA with positive cooperativity in vitro. The RNA binding activity was mapped to arginine/proline-rich motif (RPR-motif) at the C-terminus of p33 and the corresponding sequence in p92. The non-overlapping C-terminal domain of p92 also contained additional RNA-binding regions that flank the conserved RdRp motifs on both sides. Cooperative RNA binding by p33 suggested inter-molecular interactions between p33 monomers. Indeed the yeast two-hybrid and surface plasmon resonance assays revealed interactions between p33 and p33 and also between p33 and p92. The sequence involved in the protein-protein interactions was mapped to the C-terminal region in p33, proximal to RPR-motif. Within this region, mutations introduced at two short stretches of amino acid residues were found to affect p33:p33 and p33:p92 interactions in vivo and also decreased the replication of a TBSV-defective interfering RNA in yeast, a model system, supporting the significance of these protein interactions in tombusvirus replication.

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES
Memory and learning allow animals to adapt and modify their behavior towards new experiences. Some factors may result in memory deficits such as: neurodegenerative diseases, changes due to intoxication and the use of drugs. Zebrafish can be used to model complex human behavioral traits such as learning and memory. This study aims to establish a protocol for evaluating the object recognition memory or object location tasks in zebrafish. We evaluated novel object recognition memory and analyzed the exploration time of novel and familiar objects in the training and test sessions. There was a preference of the animal to explore the new object in comparison to the familiar object (61% of exploration time). We also evaluated the object location task and measured the exploration time of each object in the familiar and novel object locations. There was a preference of the animal to explore the object in the novel location in comparison to the object in the familiar location (63% of exploration time). We also evaluated the effect of the non-competitive NMDA receptor antagonist MK-801 on the object recognition and object location memory in zebrafish. In this regimen treatment, control (water only) and treated animals (5 ?M MK-801) presented a significant preference in exploring the familiar object in comparison to the new object (66 and 68% of exploration time, respectively); however, 10 ?M MK-801-treated animals did not show differences in the exploration time of the objects. In the object location task, the animals treated with the 5 or 10 ?M MK-801 did not show a significant preference for the familiar or novel location whereas the control group had a higher preference in exploring the object in the familiar location (64% of exploration time). Therefore, it is possible to suggest that 5 ?M MK-801 impaired the memory formation in an object location task, which is in agreement with previous studies demonstrating the cognitive deficit induced by MK-801 treatment in aversive and spatial memory. Considering the different responses of the control group between original task and in the regimen treatment, we evaluated the impact of habituation on cortisol levels of animals in three different protocols: 1) habituated at the experiment apparatus for 3 days (Condition 1 ? C1), 2) habituated at the experiment apparatus for 3 days plus treatment tank exposure at fourth day (Condition 2 - C2), 3) habituated at the treatment tank exposure and experiment apparatus for 3 days and exposed to treatment tank again at fourth day (Condition 3 ? C3). The results showed higher levels of cortisol in animals submitted to C2 and C3 conditions compared to animals submitted to C1. These results demonstrated that the animals submitted to treatment tank exposure have a different performance in object recognition and object location memory due to stress responses. Therefore, these tasks are prone to evaluate memory in physiological and pathological conditions, but its use is limited to perform pharmacological studies due to sensitivity to stress caused by manipulation.
A mem?ria permite que os animais possam adaptar e modificar seu comportamento diante de novas experi?ncias. Uma s?rie de fatores pode resultar num d?ficit de mem?ria como: doen?as neurodegenerativas, altera??es decorrentes de intoxica??es e uso de f?rmacos. O peixe-zebra vem sendo usado para modelar caracter?sticas comportamentais humanas complexas, como o aprendizado e mem?ria. Este estudo tem como objetivo estabelecer um protocolo para avaliar tarefas de mem?ria de reconhecimento de objetos ou de localiza??o de objetos no peixe-zebra. N?s avaliamos a mem?ria de reconhecimento de objeto novo e analisamos o tempo de explora??o de objetos novos e familiares nas sess?es de treino e teste. Houve uma prefer?ncia do animal em explorar o novo objeto em compara??o com o objeto familiar (61% do tempo de explora??o). Tamb?m avaliamos a tarefa de localiza??o de objeto e medimos o tempo de explora??o de cada objeto nos locais familiares e novos. Houve uma prefer?ncia do animal para explorar o objeto na localiza??o nova em compara??o com o objeto na localiza??o familiar (63% do tempo de explora??o). Tamb?m avaliamos o efeito do antagonista n?o competitivo do receptor NMDA MK-801 na mem?ria de reconhecimento de objeto novo e na localiza??o de objeto em peixe-zebra. Neste tratamento, o controle (somente ?gua) e os animais tratados (MK-801 5 ?M) apresentaram uma prefer?ncia significativa na explora??o do objeto familiar em compara??o com o novo objeto (66 e 68% do tempo de explora??o, respectivamente); no entanto, animais tratados com MK-801 10 ?M n?o mostraram diferen?as no tempo de explora??o dos objetos. Na tarefa de localiza??o de objeto, os animais tratados com MK-801 5 ou 10 ?M n?o mostraram prefer?ncia significativa pelo objeto na localiza??o familiar ou nova, enquanto que o grupo controle teve uma prefer?ncia maior em explorar o objeto na localiza??o familiar (64% de tempo de explora??o). Portanto, sugere-se que o MK-801 5 ?M prejudicou a forma??o da mem?ria em uma tarefa de localiza??o de objeto, o que est? de acordo com estudos pr?vios, demonstrando o d?ficit cognitivo induzido pelo MK-801 em mem?rias aversivas ou espaciais. Considerando as diferentes respostas do grupo controle entre a tarefa original e no tratamento, avaliamos o impacto da habitua??o nos n?veis de cortisol dos animais em tr?s protocolos diferentes: 1) habituado ao aparato experimental por 3 dias (Condi??o 1 - C1) 2) habituado ao aparato experimental por 3 dias mais a exposi??o ao aqu?rio de tratamento no quarto dia (Condi??o 2 - C2), 3) habituado na exposi??o ao aqu?rio de tratamento e no aparato experimental por 3 dias e exposto ao aqu?rio de tratamento novamente no quarto dia (Condi??o 3 - C3). Os resultados mostraram n?veis mais elevados de cortisol em animais submetidos a condi??es C2 e C3 em compara??o com animais submetidos a C1. Esses resultados demonstraram que os animais submetidos ? exposi??o ao aqu?rio de tratamento apresentam desempenho diferente nas tarefas de mem?ria de reconhecimento de objetos e de localiza??o do objeto devido ao estresse. Portanto, essas tarefas s?o adequadas para avaliar a mem?ria em condi??es fisiol?gicas e patol?gicas, mas seu uso ? limitado em estudos farmacol?gicos devido ? sensibilidade ao estresse causado pela manipula??o.

La tesi affronta tematiche relative al rilievo di un aggregato edilizio, all'analisi della consistenza e dei quadri fessurativi, ai criteri di lettura dei cinematismi e alle procedure di consolidamento. Vengono, inoltre, proposte strategie di progetto, finalizzate al riuso e all'auto-mantenimento sostenibile del medesimo sistema architettonico attraverso sequenze logiche e concatenate di decisioni operative, sulla base di esigenze, criticità e requisiti emersi.

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES
The increase of environmental pollutants sources directly affects the ecosystem and human health, mainly due to loss of soil and water quality. Among these toxic agents are included endocrine disruptors, which can cause changes in biochemical pathways and physiological changes in organisms. Studies with different forms of bisphenols, compounds present in many products of human consumption and the environment, had already demonstrated extremely adverse effects on behavior and gene expression of targets related to neural and endocrine systems during development. The zebrafish, Danio rerio, is an excellent model organism for studying neurotoxicity, due to its advantages with respect to specific aspects of their development and a low maintenance cost when compared to other vertebrate model organisms. This work proposes the investigation of the effects of bisphenol A (BPA), its chlorinated derivatives and its substitutes (BPS and BPF), toxicological and behavioral parameters combined with characterization of its potential underlying cellular and molecular mechanisms such as cell death and proteins and gene expression in the nervous and endocrine systems. After the embryo and larvae were exposed to different concentrations of BPA and its derivatives, morphological analyzes and embryo survival were carried out, as well as behavioral tests of locomotion and cognitive evaluation of the larvae. In the concentrantion curve of the different bisphenolic compounds, no significant difference was observed in the survival, teratogenesis and hatching of the animals, however, a significant increase was detected in the exploratory parameters of the BPA and BPF groups. When comparing the aversive behavior of the BPA with the DMSO, there was no statistical difference between the groups, whereas in the BPS an improvement in the escape response of the larvae occurred. Additionally, behavioral and locomotor activity analyzes will be performed in adult animals previously treated with the compounds studied during the initial development. In this way, it is expected to contribute to the current knowledge of neurotoxicity mechanisms related to endocrine disruption caused by contaminants of the bisphenol family.
O crescimento das fontes de poluentes ambientais afeta diretamente o ecossistema e a sa?de humana, principalmente devido ? perda da qualidade do solo e da ?gua. Entre estes agentes t?xicos, est?o inclu?dos os disruptores end?crinos, os quais podem acarretar diversas mudan?as em vias bioqu?micas e altera??es fisiol?gicas em organismos. Estudos com diferentes formas de bisfen?is, compostos presentes em diversos produtos de consumo humano e inclusive no ambiente devido ? contamina??o por efluentes industriais, j? apresentaram efeitos extremamente nocivos na regula??o da express?o de genes relacionados ao sistema neural e end?crino durante o desenvolvimento, al?m de altera??es comportamentais severas. O zebrafish, Danio rerio, ? um excelente organismo modelo para estudar neurotoxicidade, uma vez que possui vantagens em rela??o a aspectos espec?ficos do seu desenvolvimento e um custo reduzido de manuten??o quando comparado a outros vertebrados modelos. Este projeto prop?s o estudo dos efeitos do bisfenol A (BPA), seus derivados clorados e substitutivos (BPS e BPF), sobre par?metros de embriotoxicidade e comportamento, buscando tamb?m caracterizar os mecanismos celulares e moleculares dos potenciais efeitos observados avaliando morte celular e n?veis de prote?nas e express?o de genes alvos em embri?es e larvas de zebrafish. Para tanto, ap?s a exposi??o dos embri?es e larvas a diferentes concentra??es de BPA e seus derivados, foram realizadas an?lises morfol?gicas e sobreviv?ncia dos embri?es; ensaios comportamentais de locomo??o e avalia??o cognitiva das larvas. Ao realizar a curva de concentra??o dos diferentes compostos bisfen?licos, n?o foi observada diferen?a significativa na sobreviv?ncia, teratog?nese e eclos?o dos animais, por?m, ao analisar o comportamento explorat?rio dos animais tratados com BPA e BPF foi detectado um aumento significativo nos par?metros explorat?rios das larvas. Ao compararmos o comportamento aversivo do BPA com o controle tratado com o ve?culo DMSO, n?o existiu diferen?a estat?stica entre os grupos, enquanto no BPS ocorreu uma melhora na resposta de escape das larvas. Adicionalmente, ser?o realizadas an?lises comportamentais e de atividade locomotora em animais adultos previamente tratados com os compostos estudados durante o desenvolvimento inicial. Desta forma, espera-se contribuir para o conhecimento atual dos mecanismos de neurotoxicidade relacionados ? disrup??o end?crina causada por contaminantes da fam?lia dos bisfen?is.

Syftet med denna studie är att undersöka om det finns tydliga skillnader mellan mäns och kvinnors kännedom om och användning av 24 utvalda engelska lånord hämtade från nyordslistorna åren 2010–2019. Kvantitativa och kvalitativa metoder har med hjälp av en enkätundersökning använts för att få informanter att svara på om de har kännedom om dessa engelska lånord och i så fall ange användningsfrekvensen samt i vilket sammanhang orden används. Resultatet visar att inga tydliga skillnader kan fastställas mellan mäns och kvinnors kännedom om och användning av de utvalda engelska lånorden. Däremot varierar kännedomen kring några specifika lånord där skillnader kan ses mellan könen. Slutsatsen kan dras till att kännedom om och användning av engelska lånord inte skiljer sig tydligt mellan könen, men att några märkbara skillnader kan ses i kännedom om några specifika lånord mellan könen.

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Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico
Regarding the growing number of human beings with physical and mental pathologies associated to different stressor agents, attempts are being made to validate animal models with a close phylogenetic resemblance to man, to study stress response. Callithrix jacchus has been widely used in biomedical research, including on stress, but there is scarce information in the literature about how individual and social factors modulate stressor response in this species. This study uses 4 approaches to investigate the response of male and female adult C. jacchus, under situations of stress, and in the first we show evidence of the importance of this animal as an experimental model in research involving the hypothalamus-pituitary-adrenal axis. And we investigate if sex and baseline cortisol levels modulate the behavioral and hormonal response to separation. In two additional approaches investigate if type of social support (co-specific parent or non-parent) and social rank interfere in behavioral and hormonal when the animal are exposure to a new environment, paired with a co-specific (F2), exposure of the animal to a new environment, isolated (F3) or during reunion (F4). Finally, we also investigated the androgen levels in the males, with a focus on the challenge hypothesis, referring to environmental responsiveness and male-male exposure to relatives and non-relatives of C. jacchus. It was observed that: (1) the baseline cortisol of the animal is predictive of cortisol reactivity at separation; (2) males and females do not show dimorphism in the response of cortisol to stressors, although the females have higher baseline levels of this hormone and exhibit higher frequencies of anxiety-related behaviors; (3) only social support provided by relatives proved to be effective in buffering the cortisol response. In behavioral terms this response was dimorphic, showing that only the male dyads displayed an attenuated response to stress; (4) the males showed differences in cortisol levels as a function of social rank and study phases, whereas in the females no such alterations were observed. The males with indefinite dominance hierarchy (IDH) had reduced cortisol in F2 and F4, while the IDH females showed an increase in F3 and F4; (5) the males of relative and non-relative dyads did not exhibit variations in androgen levels as a function of a new environment. These results, taken together, (a) corroborate the use of C. jacchus as a good animal model for stress-related studies, given that they exhibit similar behavioral and physiological alterations to those of human beings in response to stressor agents; (b) point to the importance of considering individual and social modulating factors during experiments with stressors; (c) provide more reliable comparison parameters in studies where these primates are used as animal models, and (d) show that androgens vary as a function of genetic proximity (relative or non-relative) when the animals are faced with physical and social environmental challenges, thus providing important information for studying the challenge hypothesis in this species
Diante do aumento crescente de patologias de natureza f?sica e mental associadas aos diferentes agentes estressores, nos seres humanos, se procura validar modelos animais com maior proximidade filogen?tica do homem, para estudo da resposta ao estresse. Callithrix jacchus vem sendo amplamente utilizado em pesquisas biom?dicas, inclusive sobre estresse, mas h? escassez de dados na literatura sobre como fatores individuais e sociais modulam a resposta a estressores nesta esp?cie. Este trabalho compreende 4 abordagens de estudo da resposta de C. jacchus, machos e f?meas, adultos, em contextos de estresse. A primeira apresenta evid?ncias sobre a import?ncia desta esp?cie como modelo experimental em pesquisas que envolvam o estudo do eixo hipot?lamo-pituit?ria-adrenal. E ainda investiga se o sexo e os n?veis basais de cortisol modulam a resposta comportamental e hormonal a separa??o. Em outras duas abordagens investiga se o tipo do suporte social (co-espec?fico parente ou n?o-parente) e o posto social interferem na resposta comportamental e hormonal quando os animais s?o expostos a um ambiente novo, pareado com co-espec?fico (F2); expostos a um ambiente novo, isolados (F3) e quando reunidos (F4). Finalmente, a quarta abordagem investigou nos machos os n?veis de andr?genos, com foco na hip?tese do desafio, referentes ? responsividade ambiental e ? exposi??o macho-macho, de animais parentes e n?o-parentes. Encontrou-se que: (1) o cortisol basal do animal ? preditivo da reatividade do cortisol ? separa??o; (2) machos e f?meas n?o apresentam dimorfismo na resposta do cortisol aos estressores, embora as f?meas apresentem maiores n?veis basais deste horm?nio e apresentam maiores freq??ncias de comportamentos indicadores de ansiedade; (3) apenas o suporte social provido por parentes mostrou-se eficaz em tamponar a resposta do cortisol. Em termos comportamentais esta resposta foi dim?rfica, mostrando que apenas as d?ades de machos aparentados apresentaram uma resposta atenuada ao estresse; (4) os machos apresentaram diferen?as nos n?veis de cortisol em fun??o do posto social e das fases do estudo, enquanto nas f?meas n?o foram observadas altera??es nestas vari?veis. Enquanto os machos com hierarquia de domin?ncia indefinida (HDI) reduziram o cortisol nas fases F2 e F4, as f?meas HDI elevaram em F3 e F4; (5) os machos de d?ades parentes e n?o-parentes n?o variaram os n?veis de andr?genos em fun??o do ambiente novo. Apenas os machos parentes elevaram os n?veis de cortisol durante a fase de reuni?o, apresentado a responsividade macho-macho maior do que aquela dos n?o-parentes. Estes resultados, no seu conjunto, (a) corroboram a utiliza??o de C. jacchus como um bom modelo animal para estudos relacionados ao estresse, por apresentarem altera??es comportamentais e fisiol?gicas semelhantes ?quelas dos seres humanos em resposta aos agentes estressores; (b) apontam a import?ncia de se considerar fatores moduladores individuais e sociais durante experimentos com estressores, (c) fornecem par?metros de compara??o com aumento da confiabilidade dos estudos nos quais estes primatas s?o utilizados como modelo animal e (d) evidenciam que os andr?genos variam em fun??o da proximidade gen?tica (parente ou n?o-parente) frente a desafios do ambiente f?sico e social, fornecendo informa??es importantes para estudo da hip?tese do desafio nesta esp?cie

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Funda??o de Amparo ? Pesquisa do Estado do Rio Grande do Sul - FAPERGS
Childhood maltreatment (CM) exposure, including physical, sexual and psychological abuse, as well as physical or emotional neglect, is associated to long-term effects on mental health. The individual?s earlier years of life are characterized by rapid neurobiological and psychological development; therefore, CM is considered an important risk factor for psychological impairment and increased vulnerability to mood disorders. It is well established that alterations in the stress response systems, such as the immune and neuroendocrine systems are involved in this process. Based on this, investigating the effects of CM exposure in healthy adolescents can help to identify patterns of vulnerability to mood disorders in adulthood. The main objectives are: 1) to investigate the effects of CM exposure in neuroimunoendocrine and oxidative parameters in healthy adolescents without mood disorder; 2) to analyze early life stress effects in an animal model of maternal separation (MS) in cognitive performance, immune and oxidative parameters. The first study recruited thirty healthy adolescents reporting CM and twenty-seven healthy adolescents with no history of CM as control group. Blood, plasma and hair samples were obtained from all participants. Lymphocytes were isolated and stimulated in vitro to evaluate lymphocyte subsets, Th1/Th2/Th17 cytokines, mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (Nfkb) signaling pathways as well as lymphocyte sensitivity to dexamethasone by flow cytometry. Brain-derived neurotrophic factor (BDNF) and hair cortisol were assessed with enzyme-linked immunosorbent assays (ELISA). Increased percentage of activated T cells (CD3+CD+CD25+ and CD3+CD69+) and senescent T cells (CD8+CD28- and CD4+CD28-), as well as a reduction of NK cells (CD3-CD56+), and NKT cells (CD3+CD56+) were observed in healthy adolescents exposed to CM. Also, it was observed an increase of intracellular signaling through increased phosphorylation of ERK1 / 2 and NF-?B and increased cytokine production, such as IL-2, IFN-y and IL-17 following CM, suggesting increased cellular activation. Increased hair cortisol levels along with increased lymphocyte resistance to glucocorticoids in vitro, as well as low BDNF concentrations were observed in CM, reflecting the chronic stress effects on neuroendocrine parameters. The second study showed cognitive impairment, loss of parvalbumin and increased pro-inflammatory cytokines (TNF-?) peripherally following MS. The intervention of the enriched environment was effective in reversing memory damage as well as inflammation caused by MS. In conclusion, our results suggest that CM alter neuroimunoendocrine and oxidative responses in both, animal and human models. Thus, the understanding of the underlying mechanisms by which CM modulates the central nervous system, endocrine, and immune system and how these systems interact to alter the physiological responses of children and healthy adolescents, may help to increase preventive actions against the development of chronic inflammatory diseases and mood disorders in adulthood.
A exposi??o aos maus-tratos na inf?ncia (MTI), incluindo diferentes formas de abuso e neglig?ncia, est? associada com repercuss?es em longo prazo na sa?de do indiv?duo. Os primeiros anos de vida de um indiv?duo caracterizam-se por r?pido desenvolvimento neurobiol?gico e psicol?gico, portanto os MTI s?o considerados importantes fatores de risco para danos psicol?gicos e aumento da vulnerabilidade para transtornos de humor. Acredita-se que, altera??es em mecanismos de resposta ao estresse, incluindo os sistemas neuroend?crino e imunol?gico/oxidativo, estejam envolvidas neste processo. Diante disto, investigar os efeitos da exposi??o aos MTI no adolescente saud?vel pode ajudar a identificar padr?es de vulnerabilidade para transtornos de humor na idade adulta. Os objetivos da tese apresentada s?o: 1) investigar os efeitos neuroimunoend?crinos e oxidativos da exposi??o aos MTI em adolescentes saud?veis sem transtorno de humor e 2) analisar os efeitos do estresse precoce em modelo animal atrav?s do protocolo de separa??o materna (SM) no desempenho cognitivo e par?metros imunol?gicos/oxidativos. Para avalia??o do primeiro objetivo, foram recrutados trinta adolescentes saud?veis com hist?rico de MTI e vinte e sete adolescentes saud?veis sem hist?ria de MTI como grupo controle. Amostras de sangue, plasma e cabelo foram obtidas de todos os participantes. Linf?citos foram isolados e estimulados in vitro para identificar subtipos linfocit?rios, citocinas Th1/Th2/Th17, vias de sinaliza??o intracelular MAPKs e NF-?B, bem como a sensibilidade dos linf?citos ao dexametasona, por citometria de fluxo. Ainda, os n?veis de BDNF plasm?tico e cortisol capilar foram avaliados pela t?cnica de ELISA. Os adolescentes com MTI apresentaram um aumento na porcentagem das c?lulas T ativadas (CD3+CD4+CD25+ e CD3+CD69+) e c?lulas T senescentes (CD8CD28- e CD4+CD28-), al?m de redu??o em c?lulas NK (CD3-CD56+) e c?lulas NKT (CD3+CD56+). Al?m disso, os linf?citos de adolescentes expostos aos MTI apresentaram maior resist?ncia ao glicocorticoide dexametasona in vitro. Tamb?m evidenciamos altera??es em rotas de sinaliza??o intracelular, observada por aumento na fosforila??o de ERK1/2 e NF-?B em c?lulas T CD8+, e aumento de citocinas como IL-2, IFN-y and IL-17 ap?s MTI, confirmando um perfil de ativa??o imune celular. Aumento dos n?veis de cortisol capilar juntamente com a redu??o das concentra??es de BDNF plasm?tico foi detectado nos adolescentes expostos aos MTI, refletindo os efeitos a longo prazo do estresse cr?nico em par?metros neuroend?crinos. Ainda, um desequil?brio entre os marcadores de dano oxidativo e defesas antioxidantes foi encontrado nos adolescentes com hist?rico de MTI. Em rela??o ao segundo objetivo, os resultados do estudo experimental demonstraram que a SM causou altera??es de mem?ria, perda de parvalbumina e aumento de citocinas pr?-inflamat?rias (TNF-?) no plasma deanimais adolescentes. Ap?s a interven??o do ambiente enriquecido foi poss?vel reverter os preju?zos de mem?ria e a inflama??o causados pela exposi??o a SM. Concluindo, nossos resultados indicam que a exposi??o ao estresse durante a inf?ncia altera a resposta neuroimunoend?crina e oxidativa em modelo animal e humano. Deste modo, compreender de que forma os MTI modulam o sistema nervoso, end?crino e imune e como estes sistemas interagem entre si para alterar precocemente as respostas fisiol?gicas da crian?a e do adolescente saud?vel pode auxiliar no aumento de a??es preventivas contra o desenvolvimento de doen?as inflamat?rias cr?nicas e transtornos de humor na idade adulta.

Master of Science
Department of Animal Sciences and Industry
Evan C. Titgemeyer
Seven ruminally cannulated heifers approximately 225 kg initial BW were used in a 7 × 4 Youden square design to determine the effects of different levels of limonene on ruminal Fusobacterium necrophorum populations. Treatments included: 1) control, 2) limonene at10 mg/kg diet DM, 3) limonene at 20 mg/kg diet DM, 4) limonene at 40 mg/kg diet DM, 5) limonene at 80 mg/kg diet DM, 6) CRINA-L (a blend of essential oil components) at 180 mg/kg diet DM, 7) tylosin at 12 mg/kg diet DM. Each period included 11 d with 10 d washouts between periods. Samples were collected on d 0 (before treatment initiation), 4, 7, and 10 for measuring F. necrophorum by most probable number (MPN) method using selective culture medium. Results indicate that CRINA-L (P = 0.52) and tylosin (P = 0.19) did not affect ruminal F. necrophorum populations. Limonene linearly decreased (P = 0.03) F. necrophorum populations, and the optimal dietary concentration for limonene was 40 mg/kg DM. Limonene did not affect ruminal degradation rate of lysine, NH3 concentration, or VFA profiles in ruminal fluid. Limonene was useful for reducing ruminal concentrations of F. necrophorum. It may have potential to control liver abscesses, although further research will be needed to assess the effect of limonene under feedlot conditions.

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Os programas de transfer?ncia condicionada de renda est?o disseminados em pa?ses em desenvolvimento na Am?rica Latina com ?nfase no Brasil como um novo paradigma nas pol?ticas sociais para erradica??o da pobreza. Consistem no repasse de recusros monet?rios de renda do governo diretamente para familias pobres mediante cumprimento das condicionalidades em educa??o e sa?de. Na sa?de, memos com vari??oes entre os pa?ses as condicionalidades tem como p?blico alvo gestantes e crian?as com vistas ? redu??o de indicadores de sa?de da crian?a, como cescimento e desenvolvimento, mortalidade infantil e incemento do pr?-natal. Os objetivos deste trabalho s?o comparar o programa de transferencia condicionada de renda brasileiro, o Bolsa Familia e os programas semelhantes na Am?rica Latina em rela??o aos seus efeitos no crescimento e desenvolvimento de crian?as at? 7 anos de idade e avaliar a prevalencia das condicionalidades em sa?de da mulher e da crian?a entre usu?rias dos servi?os de sa?de brasileiros cujas equipes aderiram ao Programa de Acesso e Qualidade da Aen??o B?sica (PMAQ). Para o primeiro objetivo foi realizada uma revis?o sistem?tica onde selecionaram-se dez artigos entre mais de mil encontrados nas bases de dados Embase, Pubmed, Scopus, Scielo e Lilacs. Os artigos s?o estudos epidemiol?gicos observacionais dos tipos descritivos transversais e anal?ticos de coorte e casos-controle. Para o estudo de prevalencia, an?lise estat?stica utilizou a regress?o de Poisson com vari?ncia robusta para investigar como a preval?ncia do cumprimento das condicionalidades na ?rea de sa?de foi influenciada pelas diversas vari?veis explicativas. Foram estimadas raz?es de preval?ncias brutas e ajustadas, com seus respectivos intervalos de confian?a de 95%. O cadastro no programa bolsa fam?lia foi considerado como principal vari?vel de exposi??o. As vari?veis de confundimento foram: idade da m?e, escolaridade, ra?a/cor, saber ler, viver sozinha e regi?o de moradia. Na an?lise dos dados foi utilizado o software R 3.0.1 (RDevelopment Core Team, 2013).Em rela??o a compara??o do Bolsa Familia com demais programas da Am?rica Latina, a revis?o mostrou resultados semelhantes quanto ao efeito positivo dos programas de transfer?ncia de renda no estado nutricional de crian?as benefici?rias, sendo que estes efeitos s?o mais evidenciados em crian?as menores de dois anos de idade, bem como pertencentes ?s fam?lias de mais baixo n?vel socioecon?mico. Para a prevalencia das condicionalidades entres os diferentes grupos de usu?rias do Bolsa Fam?lia e n?o usu?rias Os resultados apontaram que n?o houve diferen?a estat?stica significativa entre respondentes (com filhos menores de dois anos) cadastrados e n?o cadastrados no PBF quanto ?s quest?es referentes aos seguintes aspectos: cumprimento do calend?rio vacinal da crian?a, realiza??o de pelo menos sete consultas de pr? natal, realiza??o do acompanhamento do crescimento e desenvolvimento da crian?a e receber orienta??o da equipe de sa?de quanto a alimenta??o da crian?a. Conlui-se de um lado que aumento de renda m?nima para familias de exterma pobreza mostrou impacto possitivo na sa?de das cran?as no Brasil e Am?rica Latina. Do outro n?o confirmou, no Brasil, uma incremento das condicionalidades expressas numa utliza??o dos servi?os de aten??o b?sica por parte das usu?rias do Bolsa Familia.
The programs of conditional cash transfer are widespread in developing countries in Latin America with emphasis on Brazil as a new paradigm in social p olicies for poverty eradication . Consist of transferring monetary funds from the government directly to poor families by fulfilling the condition alities on education and health . In health, even wi th variations between countries , conditionality targeting public pregnant women and children with a view to improving health indic ators maternoinfantil as growth , infant mortality and prenatal care. The objectives of this study are to compare the transfer progr am conditional Brazilian income , the Family and similar programs in Latin A merica in relation to it s effects on growth in children , and to evaluate the effect of Bolsa Fam?lia in the prevalence of use of services ( conditionalities ) of prenatal care in Brazilian health services whose teams joined the Programa de Acesso e Melhoria da Qualidade da Aten??o B?sica (PMAQ - AB) . For the first objective a systematic review , we selec ted ten articles between 1007 ( one thousand and seven ) found in the databases Embase , PubMed, Scopus , Scielo and Lilacs databases was performed . Articles are ob servational epidemiological studies of transverse descriptive and analytical types of cohort and case - co ntrol. For the second objective, for it is a prevalence study , a statistical analysis using Poisson regression with robust variance was performed to i nvestigate how the prevalence of compliance with conditionalities on health was influenced b y various explanatory variables . Ratios , crude and adjusted prevalence , with their respective confidence i ntervals of 95 % were estimated . The family joined the sch olarship program was considered as the main expo sure variable . Confounding variables were: maternal age , race / color, paid employment , marital status and region of residence . In d ata analysis software R 3.0.1 (RDevelopment Core Team 2013 ) was used . Rega rding the comparison of the Bolsa Fam?lia with other programs in Latin America , the review found similar results regarding the positive effect of income transfer in the nutritional status o f beneficiary children programs , and these effects are more evident in children under two years old and belonging to familie s of lower socioeconomic status . For the prevalence of conditionalities entres different groups of users of the Bolsa Fam?lia and nonusers results showed no statistically significant difference betwe en respondents (with children under two years ) registered and not registered in PBF on issues relating to: me et at least six prenatal visits , meet and participate in health education activities . It follows from side to increase minimum income for families in extreme poverty showed positive impact on children's health in Brazil and Latin America. The o ther is not confirmed in Brazil , an increase in conditionality expressed in use of primary care by the user s of the Bolsa Fam?lia services.

In a quest to design novel deployable structures, flexible polyhedra provide interesting insights. This work follows the discovery of flexible polyhedra and aims to make flexible polyhedra more useful. The dissertation describes how flexible polyhedra can be made. The flexible polyhedra first considered in this dissertation have a rotational degree of freedom. The range of this rotational movement is measured and maximised in this work by numerical maximisation. All polyhedra are established computationally: an iterative solution method is used to find vertex coordinates; several clash detecting methods are described to define whether each rotational position of a flexible polyhedron is physically possible; then a range of motion is defined between occurrences of clashes at the two ends; finally, an optimisation tool is used to maximise the range of motion. By using these tools, the range of motion of two types of simplest flexible polyhedra are maximised. The first type is a series of flexible polyhedra generalised from the Steffen flexible polyhedron. The range of motion of this type is improved to double that of Steffen’s original, from 27° to 59°. Another type of flexible polyhedron is expanded from a model provided by Tachi. Based on the understanding of Steffen’s flexible polyhedron, optimisation parameters are carefully given. This new type has achieved a wider range of motion, so now the range of motion of flexible polyhedron is tripled to 80°. After enlarging the range of motion of the degree of freedom in the 1-dof systems, the dissertation found multiple degrees of freedom in one polyhedron. The multiple mechanisms can be even repetitive, so that an n-dof polyhedron is found. A polyhedron of two degrees of freedom is first presented. Then, a unit cell for any number of mechanisms is found. As a repetitive structure, a 3-dof polyhedron is presented. Finally, this work presents the possibility of configuring a flexible polyhedral torus and a closed polyhedral surface that is able to flex without the need to stop.

Many plant RNA viruses provide replication and encapsidation functions for one or more subviral RNAs that can modulate the symptoms of the associated helper virus. In this dissertation, I report my studies on symptom modulation in Arabidopsis thaliana by subviral RNAs associated with turnip crinkle virus (TCV), a single-stranded, positive-sense, plant RNA virus. Satellite RNA C (satC) is a virulent satRNA that normally intensifies symptoms of wild-type (wt) TCV but can attenuate symptoms if the TCV coat protein (CP) is either replaced with that of cardamine chlorotic fleck carmovirus (TCV-CPCCFV) (Kong et al., 1995) or if TCV contains an alteration in the CP initiation codon (TCV-CPm) (Kong et al., 1997b). I found that TCV-CPm produced reduced level of a CP (10∼20% of wt level) that contained two additional amino acids at its N terminus and did not form virions in infected protoplasts. SatC did not substantially affect the accumulation of TCV-CPm genomic RNA in protoplasts. These results, along with data reported previously (Kong, 1996), led to the conclusion that satC-mediated symptom attenuation of TCV-CPm involves a reduction in virus long-distance movement (Kong et al., 1997b). By characterizing the promoter for the CP mRNA, i.e., the 1.45-kb sgRNA promoter and defining the sequence and structural elements required for promoter activity, I was able to construct TCV variants expressing a level of wt CP similar to TCV-CPm (10∼20% of wt). I found that these mutants also have their symptoms attenuated by satC, indicating that the level of viral CP, not the mutation in the N terminus, is the crucial factor in determining whether satC is going to attenuate or exacerbate symptoms. Another normally virulent subviral RNA, namely defective interfering RNA G (diG) exhibited different symptom modulation of TCV-CPm compared with satC, and the determinants for this differential symptom modulation were previously localized to the T-terminal 100 bases of the subviral RNAs containing six positional differences (Kong et al., 1997a). In this dissertation I report the further characterization of the determinants in these six positions and the possible mechanism underlying the differential symptom modulation by these two subviral RNAs. My results revealed that two positions located in the 3′-terminal stem-loop structures of satC and diG, which also serve as promoters for complementary strand synthesis, are critical for symptom modulation. Furthermore, the hairpin CP binding capacity correlates with the symptom modulation. Several models for symptom modulation by the subviral RNAs associated with TCV are proposed.

Replication of plus (+)-strand RNA viruses proceeds through minus (−)-strand intermediates. Satellite RNA C (satC), one of the nonessential subviral RNAs of Turnip crinkle virus (TCV), is dependent on the TCV-encoded RdRp for its replication. Earlier work showed that a stem-loop structure at the 3′ end of (+)-strand satC is required for synthesis of (−)-strands (Song and Simon, 1995a). Using an in vitro RdRp assay, I defined two separate cis-acting elements on satC (−)-strands that can promote complementary strand synthesis. One element comprises 11 bases and is located near the 3′ end (3′-proximal), and the other consists of 14 bases and is located 41 bases from the 5′ end (5′ -proximal). Both elements contain multiple consecutive C residues followed by multiple consecutive purines. In vivo mutagenesis and genetic selection (SELEX) studies were carried out to investigate the functional significance of the two elements as well as the satC (−)-strand 3′ terminus (3 ′ OH-CCCUAU), which contains the (−)-strand 3′ -end sequence 3′ OH-CC1–2(A/U)(A/U)(A/U) found in all carmovirus RNAs (named the carmovirus consensus sequence or CCS). My results indicate that the 3′-terminal CCS and the 5′-proximal element are highly conserved and required for satC (+)-strand synthesis. Although mutations introduced into the 3′-proximal element were tolerable, this element preferentially contains a sequence similar to the CCS and/or polypurines, suggesting that this element may also contribute to satC accumulation in vivo. All RNAs associated with TCV terminate with the motif CCUGCCC-3 ′ at the 3′ end. Transcripts of satC containing a deletion of the motif, or the 3′-terminal 6 bases, are nearly always repaired to wild-type in vivo by RdRp-mediated primer extension of oligoribonucleotides synthesized by abortive initiation and complementary to the 3′ end of TCV genomic RNA (Nagy et al., 1997). In this thesis, I provide evidence that two additional mechanisms are used by the TCV RdRp to repair shorter deletions of the 3 ′-end motif of satC. Deletion of the 3′-terminal CCC residues along with addition of 8 non-template bases is repaired in vivo mainly by homologous recombination between the similar 3 ′ ends of satC and TCV. Deletion of the 3′-terminal 4 or 5 bases, in the presence or absence of non-template bases, led to recovery of progeny containing a mixture of wild-type 3′ ends and non-wild-type 3′ ends that included base alterations, deletions and insertions. Assays using an in vitro RdRp transcription system indicate that the TCV RdRp is likely able to polymerize nucleotides in a template-independent, non-random fashion before initiating transcription of deletion-containing satC. The existence of 3 different repair mechanisms associated with a single virus suggests an intrinsic need for 3′ -end reconstruction in the cellular environment.

In this dissertation, I report my studies on the resistance of Arabidopsis thaliana ecotype Dijon (Di-0) to turnip crinkle carmovirus (TCV), a simple, single-stranded, positive-sense, plant RNA virus. TCV efficiently replicated in protoplasts prepared from Di-0 callus culture, but it did not move long-distance in Di-0 plants (Simon et al., 1992). Cardamine chlorotic fleck virus (CCFV), the carmovirus most closely related to TCV, infected Di-0 plants systemically (Oh et al., 1995). I found that TCV with the coat protein open reading frame (ORF) from CCFV (TCV-CP$\rm\sb{CCFV})$ and TCV with a single base mutation in the initition codon (AUG to ACG) of the coat protein ORF (TCV-CPm) were infectious on ecotypes Columbia (Col-0, TCV-susceptible) and Di-0 (TCV-resistant). These results indicate that TCV coat protein is the viral determinant for resistance of Di-0 to TCV. In addition, my results suggest that the resistance of Di-0 to TCV is specific to TCV and may involve an RNA degradation activity. I also studied the movement of TCV, CCFV, and TCV-CP$\sb{\rm CCFV}$ in Col-0 and Di-0 plants using a whole plant in situ hybridization technique. The results indicate that TCV was restricted to within small areas around the initial infection sites on inoculated leaves of Di-0 plants, whereas TCV (in Col-0), CCFV and TCV-CP$\sb{\rm CCFV})$ (in Col-0 and Di-0) moved long-distance to metabolic sink tissues (young leaves and roots) at different rates. Many plant RNA viruses are associated with small subviral RNAs, including satellite (sat-) RNAs. Small subviral RNAs require a helper virus for replication and movement, and they often intensify or attenuate the symptoms caused by the helper virus. TCV is associated with sat-RNA C, a sat-RNA that normally intensifies the symptoms of the TCV-M isolate (Simon et al., 1988; Li and Simon, 1990). I will report my work on the mechanism of symptom modulation mediated by sat-RNA C in Arabidopsis plants. My results indicate that symptom modulation by sat-RNA C is mediated by the viral coat protein, and a putative interaction between the 3$\sp\prime$ end of sat-RNA C and the N-terminus of coat protein is involved. Symptom attenuation by sat-RNAs is widely believed to be mediated by inhibition of helper virus replication through competition for replication factors (Roossinck et al., 1992). My study shows that inhibition of virus long distance movement is involved in sat-RNA C-mediated symptom attenuation of TCV-CPm (and probably TCV-CP$\rm\sb{CCFV})$ and inhibition of helper virus replication is not important. In addition, symptom attenuation mediated by sat-RNA C is localized and does not involve the major plant defense pathway. A model that explains sat-RNA C-mediated symptom attenuation is proposed.

In this dissertation, I report the functions of the TCV coat protein in TCV viral and subviral RNA replication using in vivo and in vitro systems. In chapter 2, I present my results on construction of a biologically active, full-length cDNA of cardamine chlorotic fleck virus (CCFV), a new carmovirus. In chapter 3, I show that p28 and p88 of TCV viral RdRp components can be expressed in insect cells using a baculovirus expression system. In addition, I demonstrate that p28 and p88 are present in infected plant cells using antiserum against p28 fusion protein expressed in E. coli. In Chapter 4, using a TCV derivative with a deletion of coat protein ORF (TCV$\Delta$CP) and several other TCV derivatives that alter the TCV coat protein ORF, I show that TCV coat protein up-regulates the accumulation of sat-RNA C and DI RNA G, but not sat-RNA D, in protoplasts, suggesting that TCV coat protein plays a role in regulation of subviral RNA replication in vivo. In chapter 5, I show that TCV coat protein functions in TCV viral and subviral RNA transcription and TCV RdRp expression. The immunochemical analysis of cellular proteins obtained from protoplasts infected with TCV$\Delta$CP, TCV, TCV-CP$\rm\sb{CCFV}$ (a TCV derivative containing the CCFV coat protein ORF, or TCV-CPm (a TCV derivative producing low amounts of TCV-related coat protein) suggests that TCV coat protein might be involved in the repression of TCV viral RdRp synthesis and regulation of readthrough of p28, leading to a change in the stoichiometry of p28 and p88. I show evidences that TCV RdRp prepared in the absence of coat protein has template dependency and specificity with small, subviral RNA sized-templates. However, with genomic RNA sized (3-4 kb) templates, no strict template specificity is observed. Using an in vitro RdRp assay system free of coat protein, I demonstrate that TCV coat protein not only enhances the transcription of minus- and plus-strand RNA of sat-RNA C and DI RNA G at low concentrations, but also inhibits the transcription of both strands of sat-RNA C and DI RNA G at higher concentrations. Using sat-RNA D minus-strand template, no inhibition of transcription is observed even in the presence of higher than 10-fold molar ratio of TCV coat protein to template. Similar stimulatory and inhibitory effects of TCV coat protein on the transcription of TCV plus- and minus-strand genomic RNA are observed, and the inhibitory effect is more pronounced during minus-strand synthesis. These in vitro RdRp assay results suggest that the TCV coat protein may act as an important viral factor involved in regulation of viral and subviral RNA replication during virus infection in plants.

碩士
國立中興大學
植物病理學系
84
Sweet pepper plants grown in the fields with leaf showing symptoms of crinkle and mottle were collected as the studing material. Three isolates ( L1, L2 and L3 ) were isolated with different local lesion indicator. Identification of the three virus isolates was further conducted by host reactions, physical properties, serological tests and electron microscopy. All of these three virus isolates had the same characteristics. They caused yellow ring spots on Chenopodium amaranticolor, C. quinoa, Datura metel and same system symptoms on other indicator plants, their thermal incativation point were 60~70℃, dilution end point were 10-6~10-7 , and the particles were all similar as 540~560 nm × 13nm. According to the same properties, these three isolates from Taiwan were conduced as the same strain of potato virus X. The titers of the antisera produced from rabbit by injecting purified virus as antigen was 1024 as determined by the interface precipitin test. Two different enzymes ( alkaline phosphatase and horse-radish peroxidase ) were conjugated to the fractionated antibody for the use in DAS-ELISA ( double antibody sandwich immunosorbent assay ) to compare the efficiency on detecting virus. When the peroxidase inhibitor such as NaN3 or H2O2 in ethanol was added to the plant crude extract with 30 min before loading the enzyme conjugated antibody in DAS-ELISA processing, the background reaction was reduced. HRP-antibody conjugated by periodate-oxidation precisely detected the virus at the concentration of 1: 2000 and the reations were easily visulized after the treatment with adding peroxidase inhibitors such as NaN3 and H2O2 in ethonal before loading the HRP conjugate antibody.

Este relatório de Prática de Ensino Supervisionada contempla as aprendizagens feitas ao longo do ano letivo 2013/2014 no âmbito do Mestrado em Educação Pré-Escolar. Encontra-se dividido em duas partes, a Parte I, relativa à Prática de Ensino Supervisionada em contexto Creche e a Parte II, alusiva à Prática de Ensino Supervisionada em contexto Jardim de Infância. A Parte I está dividida em dois capítulos. O Capítulo I é constituído por uma dimensão reflexiva sobre as aprendizagens realizadas em contexto Creche e o Capítulo II apresenta o ensaio investigativo sobre as interações das crianças com os materiais nos momentos de brincadeira livre desenvolvido com duas crianças, numa lógica de investigação qualitativa. Os resultados revelaram que as crianças participantes do estudo estabeleciam diferentes interações com os materiais, numa incessante descoberta do que as rodeava, observando, explorando e fazendo experiências no seu mundo. A Parte II, relativa ao contexto de Jardim de Infância, encontra-se, também, dividida em dois capítulos. No Capítulo III é apresentada uma reflexão sobre as aprendizagens realizadas neste contexto e no Capítulo IV é apresentado o projeto intitulado “Os ovos” desenvolvido segundo a Metodologia de Trabalho de Projeto com 15 crianças. Com o desenvolvimento deste projeto as crianças tiveram oportunidade de aprender sobre os ovos e de cooperar umas com as outras, aprendizagens que se situam na área de formação pessoal e social, de conhecimento do mundo e de expressão e comunicação (Ministério da Educação, 1997).

碩士
國立中正大學
會計資訊與法律數位學習碩士在職專班
105
There are so many human rights protection issues related to police duties, including personal freedom, rummage, interrogation, detention and transfer of criminal……etc. These duties are clarified in the Con-stitution, being discussed in the Constitutional Court and also being found to have some academic discussions. The Police in the representative of the country perform the public authorities including interference the execution, banning and administrative enforcement. Without care, the power might violate the human right easily. The police are the first line law-executor. Along with the rise of civic law literacy, the police not only have to deal with the various cases of public security and traffic accident, but also have to protect people's rights. Especially, when point-policeman are directing the traffic, maintaining the public security, and cracking down the danger drivers, their behaviors might have lots of to do with people. Dealing with the issues concerning the traffic offenders` arguments, the claims of legitimate of law practice, and even state compensation during the services, are now unavoidable topics for the police. Base on the Constitution in the democracy country and the principle of executing by law, people's rights are free from government`s unreasonable interference. Without the law and the authorized orders as the basis, administrative organization shall not interfere people`s rights. Furthermore, even if“the administrative discretion right” permits administrative organizations can judge independently, different policemen can make different decisions in the same event or even the same police could make different decisions to different people at the roughly same situation though. Of course, in police education, police had been taught repeatedly that under the administrative discretion right, police should abide by "the principle of proportionality," ”principle of equality before the Law “ “principle of legitimate expectation” in order not to violate people`s rights. But in practice, it is really difficult to set the standard of the relative required handling techniques, and how to address the specific situation of law disputes. In this study, we focus on how police deal with those cases that people did not abide the law and put themselves in dangers first, and then got hurt due to their carelessness and what responsibilities the police should take. We study about how the police can react in similar situation. As a result, we take how the point-policemen being treat in the traffic accidents as the reference to analysis what elements can lead to state compensation events caused by civil servants` works, the association between point-police service and the state compensation, circumstance of service execution, the law basis, legal facts of the case, the pros and cons of lack of standard operating procedures. All in all, we make the final recommendations that the National Police Agency should improve the "SOP, Standard Operating Procedure," revise the law of state compensation, face the difficulties the police have while executing service, we hope that the judge will respect police`s discretion right, and let "law" "reason" "love", “favoring” being considered under administrative ethics. Keywords: Accurate process of law, state compensation, administrative discretion, SOP standard operating procedures, the principle of proportionality, the principle of equality before the law, the principle of legitimate expectation.

Rhabdoviridae family viruses attacking the plant hosts were only described at the angiosperms. In this work, a gymnosperm rhabdoviridae infection was described for the first time ever ? specifically at Ginkgo biloba. Even though there were not observed any kinds of obvious infection symptoms on any of randomly chosen plant samples, through the molecular methods and detection primers the rhabdoviridae infection was proved at six of the plant samples. The acquired nucleotide and amino acid sequences, which were compared with the GenBank sequences, confirm the Rhabdoviridae family viruses occurrence. These given sequences demonstrated a certain analogy with a Strawberry crinkle virus assigned to the genus of Cytorhabdoviruses. The analyses proved a different reciprocal homology among the nucleotide sequences of the individual isolates, and in all likelihood an occurrence of two up to now unknown viruses in the Ginkgo biloba samples was proved for the first time. A definite categorization will be dependent on an acquisition and comparison of other sequences from the isolates genome and also on certain biological characteristics observation.

Notre mémoire se veut une lecture de la version américaine de la série à succès The Office (NBC, 2005-2013) à travers la métaphore dramaturgique du sociologue Erving Goffman (1922-1982). Ce rapprochement permet une riche analyse, puisque la série comme le penseur placent le malaise au cœur de leurs projets respectifs. En ayant comme prémisse fictive d’être un documentaire, ainsi qu’en se faisant une satire de l’auto-spectacularisation des participants de télé-réalité, The Office explicite la métaphore dramaturgique goffmanienne. Les personnages de la série, et particulièrement le protagoniste Michael Scott, rendent constamment sensible le fait qu’ils sont en représentation. Le malaise étant une émotion fondamentalement désagréable, la série semble désigner les règles qu’elle considère comme essentielles à la cohésion du groupe en nous faisant subir leur transgression. Le malaise que nous éprouvons deviendrait l’occasion pour nous d’expérimenter les conséquences sociales de transgressions, nous apprenant du même coup l’importance de respecter les normes. D’abord, nous essayerons de déterminer si la série permet quelque chose comme un apprentissage par la négative des règles sociales. Dans un deuxième temps, nous tenterons de prouver que certains épisodes présentent un discours différent de celui de Goffman sur la transgression des normes sociales, présentant des moments où le malaise peut être vécu par les personnages comme des opportunités d’approfondir certaines relations interpersonnelles ou de faire des gains politiques.
Our study is intended as a reading of the American version of the successful series The Office (NBC, 2005-2013) through the dramaturgical metaphor of sociologist Erving Goffman (1922-1982). This rapprochement allows for a rich analysis as both the series and the sociologist place social embarrassment at the heart of their respective projects. With the fictional premise of being a documentary, as well as satirizing the self-spectacularization of reality TV participants, The Office makes Goffmanian dramaturgy explicit. The characters in the series, especially protagonist Michael Scott, constantly make us sensitive to the fact that they are in performance. Embarrassment being a fundamentally unpleasant emotion, the series seems to point to the rules it sees as essential to group cohesion by making us suffer their transgression. The discomfort we feel could become an opportunity for us to experience the social consequences of transgressions, teaching us the importance of upholding norms. First, we'll try to determine if the series allows for something like "negative learning" of social norms. Second, we will try to prove that some episodes convey a different narrative than Goffman's on the transgression of social norms, presenting moments when embarrassment can be experienced by the characters as opportunities to deepen some interpersonal relationships or to make political gains.