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Microarrays for the scalable production of metabolically relevant tumour spheroids: a tool for modulating chemosensitivity traitsHardelauf, Heike, Frimat, Jean-Philippe, Stewart, Joanna D., Schormann, Wiebke, Chiang, Ya-Yu, Lampen, Peter, Franzke, Joachim, Hengstler, Jan G., Cadenas, Cristina, Kunz-Schughart, Leoni A., West, Jonathan 02 April 2014 (has links) (PDF)
We report the use of thin film poly(dimethylsiloxane) (PDMS) prints for the arrayed mass production of highly uniform 3-D human HT29 colon carcinoma spheroids. The spheroids have an organotypic density and, as determined by 3-axis imaging, were genuinely spherical. Critically, the array density impacts growth kinetics and can be tuned to produce spheroids ranging in diameter from 200 to 550 µm. The diffusive limit of competition for media occurred with a pitch of ≥1250 µm and was used for the optimal array-based culture of large, viable spheroids. During sustained culture mass transfer gradients surrounding and within the spheroids are established, and lead to growth cessation, altered expression patterns and the formation of a central secondary necrosis. These features reflect the microenvironment of avascularised tumours, making the array format well suited for the production of model tumours with defined sizes and thus defined spatio-temporal pathophysiological gradients. Experimental windows, before and after the onset of hypoxia, were identified and used with an enzyme activity-based viability assay to measure the chemosensitivity towards irinotecan. Compared to monolayer cultures, a marked reduction in the drug efficacy towards the different spheroid culture states was observed and attributed to cell cycle arrest, the 3-D character, scale and/or hypoxia factors. In summary, spheroid culture using the array format has great potential to support drug discovery and development, as well as tumour biology research. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.
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Sexual selection in Drosophila simulansSharma, Manmohan Dev January 2010 (has links)
Over the last 100 years sexual selection has advanced into a vast field of theoretical and empirical research. While Darwin’s idea of female preference being an integral mechanism of sexual selection is no longer debated, our understanding of female preference is still very limited. For example, we know little about the genetic variation in female preference, and the costs of preference over and above the costs of mating with particular male phenotypes. Additionally, while costs of mate choice are well documented, the benefits of mate choice and their implications are still debated. For example, controversy exists over the inevitability of good gene benefits and their capability to promote adaptive sexual selection. Furthermore, the adaptiveness of sexual selection itself is debated. Our understanding of the traits involved in mate choice is also far from complete. Here I investigated aspects of sexual selection in Drosophila simulans, employing a range of behavioural approaches along with artificial selection and environmental manipulations. The findings presented here indicate that female preference can evolve when directly selected on, and that preference itself is not particularly costly. There was also no conclusive evidence for the good genes benefits of mate choice in D. simulans. These benefits are considered crucial in promoting the adaptiveness of sexual selection, and although we found sexual selection to be adaptive under some test conditions it was not adaptive in other conditions. Our investigations into traits involved in mate choice established sex-specific genetic variation in cuticular hydrocarbons and the genetic architecture of this trait was found to sex-specific evolution of cuticular hydrocarbons under natural and sexual selection. Additionally, we found that a secondary sexual character, the sex combs was positively allometric – just like most signalling and weapon traits, and there was no association between trait fluctuating asymmetry and trait size. These findings collectively indicate that sexual selection in D. simulans is consistent with classical models of this process.
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Interactions plante - pollinisateur : caractérisation de la qualité du pollen de deux cucurbitacées durant son ontogenèse, sa présentation et son transport sur le corps de l'abeille domestique / Plant – pollinator interactions : characterisation of pollen quality during its ontogenesis, its presentation and its transport on honey bee body in two cucurbitaceaeDibos, Chloé 14 December 2010 (has links)
Chez de nombreuses Angiospermes, la pollinisation croisée est nécessaire pour le succès de la reproduction. La plupart de ces plantes ont évolué afin de favoriser la pollinisation entomophile, principalement assurée par les abeilles. C’est le cas des Cucurbitacées, dont le melon (Cucumis melo L.), importante culture du sud de la France et le concombre d’âne (Ecballium elaterium (L.) A. Rich.) plante spontanée endémique du bassin méditerranéen, les deux modèles de cette étude. Afin de mieux comprendre les relations plante-pollinisateur,nous avons choisi de caractériser le couple pollen de Cucurbitacées/abeille domestique (Apis mellifera L.) à travers la production de pollen, sa présentation sur la fleur et son transport sur l’abeille. Nous avons montré que, chez ces plantes, l’ontogenèse du pollen s’accompagne d’erreurs développementales conduisant à la formation de près de 3% de grains de pollen vides. De plus, nous avons mis en évidence que chez E. elaterium, des erreurs méiotiques entraînaient la formation de grains de pollen génétiquement anormaux qui pourraient être source d’autopolyploïdie. Chez C. melo, nos résultats ont révélé que la mise en contact du pollen avec le milieu environnant entraîne une baisse de 30% de sa viabilité, mais que celle-ci reste stable jusqu’à la fin de l’anthèse. L’aptitude à germer, quant à elle, finit d’être acquise juste avant anthèse et décroît de 12% en fin de période d’anthèse. Nous avons montré que le pollen de C. melo transporté sur le corps de l’abeille pouvait soit avoir une viabilité et une aptitude à germer préservée, probablement par protection contre la déshydratation, soit perdre quasi-totalement sa viabilité et son aptitude à germer. Enfin, nous avons détecté des composés biochimiques spécifiques des abeilles protégeant ou diminuant la qualité reproductrice du pollen / For many flowering plant species cross pollination is necessary to ensure reproductive success. Most of these plants have evolved to encourage insect pollination which is mainly carried out by bees. Such is the case of plants in the Cucurbitaceae family, including the cantaloupe (Cucumis melo L.), an important crop of the South of France, and squirting cucumber (Ecballium elaterium (L.) A. Rich.), a wild plant restricted to the Mediterranean Basin, the two vegetal models used in this study. To better understand plant-pollinator relations, we proposed to characterise the interaction between Cucurbitaceae pollen and the honey bee (Apis mellifera L.) through pollen production, its presentation and its transport on the honey bee body. Our results showed that developmental anomalies took place during pollen ontogenesis in these species leading to 3% of the pollen grains produced to be empty. Moreover, we demonstrated that meiotic abnormalities in E. elaterium leading to the production of genetically abnormal pollen grains could be a source of autopolyploidy. At anthesis in C. melo, our results showed that pollen viability decreased to 30% when pollen was exposed to environmental conditions, then remained stable during the period of anthesis.The ability for pollen to germinate was completed just before anthesis then decreased to 12% at the end of anthesis. We determined that viability and germinability of cantaloupe pollen carried on the honey bee body can be enhanced or decreased according to which specific honey bee biochemical compounds were detected
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Characterisation of engineered nanoparticles and their interaction with natural biological and non-biological materialTaylor, Cameron S. January 2014 (has links)
Form, mobility, toxicity and the eventual fate of engineered nanomaterials in environmental ecosystems are currently not well defined and are needed to improve risk assessment and legislation. The present study subjected uncoated zinc oxide (ZnO) nanoparticles (30nm and 200nm) and coated silver (Ag) nanoparticles (Paraffin: 3-8nm and citrate/PVP: 50nm) to different ionic strength media and different types of algal/bacterial extracellular-polymeric species (EPS) at long (6 months) and short (2 weeks) timescales. Changes in particle size distribution and stability were examined using a multi-method approach. Sample concentration and sample polydispersity are important factors when selecting techniques. Uncoated ZnO nanoparticles aggregated heavily in water at high concentrations (1000mg/L). However silver nanoparticles (1-10mg/L) remained stable at all ionic strengths and EPS in this study due to the steric component of their coatings. Nano-toxicological experiments involving cyanobacteria S.leopoliensis and green algae C.reinhardtii showed size-dependent toxicity from coated nanosilver particles. Smaller nanoparticles (3-8nm) showed greater dissolution over 72h and greater toxicity to both species than 50nm particles indicating silver ions are an important toxicity mechanism. Nanoparticle coatings were likely important in controlling dissolution levels. Cell viability and production of reactive oxygen species (ROS) were shown to be important mechanisms of toxicity to phycological species. Species specific effects were noted for both silver nanoparticles. EPS from S.leopoliensis were noted to remove ionic silver from suspension and different types of C.reinhardtii EPS were produced when particles underwent different levels of toxic stress indicating that EPS could both affect particle toxicity and be affected by it. This work has demonstrated that coated nanoparticles could remain stable under various ionic strengths and with exposure to algal organic matter for timescales up to 6 months. This could result in adverse effects to aquatic organisms were they to reach environmental systems and is of concern to nanomaterial risk assessors.
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Étude de l'impact de combinaisons d'acides gras et de l'insuline sur la fonctionnalité des cellules musculaires lisses vasculairesSt-Denis, Corinne 06 1900 (has links)
L’athérosclérose est étroitement liée au diabète de type 2. De fortes concentrations plasmatiques en acides gras libres (AGL) et en insuline sont des caractéristiques retrouvées chez les patients souffrant de ces deux pathologies. Les AGL, présents dans notre alimentation, font partie de l’environnement auquel les cellules sont exposées. Leurs effets dépendent de leur nature, les acides gras saturés (AGS) étant néfastes et les acides gras monoinsaturés (AGMI) plus protecteurs. Ils ont donc des effets variés sur les cellules musculaires lisses vasculaires (CMLV) impliquées dans la pathogénèse de l’athérosclérose.
Ainsi, l’objectif principal de ce projet de maîtrise était d’évaluer l’impact de deux combinaisons d’AGL sur la viabilité des CMLV, en condition hyperinsulinémique ou non. Les deux combinaisons renfermaient les mêmes AGL mais en proportions différentes, l’une étant plus riche en AGS et l’autre en AGMI. Nos résultats ont montré que les combinaisons d’AGL ont un effet pro-apoptotique principalement dû aux AGS. L’acide oléique présent dans les combinaisons atténue cependant cet effet. Il diminue même plus fortement l’apoptose des CMLV lorsqu’associé à un AGS que lorsqu’utilisé seul. Cet impact est significatif uniquement dans certaines proportions de ces AGL et est plus efficace en présence d’insuline.
Ces résultats mettent en lumière la présence d’une compétition entre mécanismes anti- et pro-apoptotiques en fonction des proportions d’AGS versus AGMI et de l’insulinémie chez les CMLV. Ils soulignent également l’importance de la présence des AGMI dans les diètes riches en AGS et pourraient être utiles pour l’élaboration de nouvelles diètes adaptées aux patients athérosclérotiques et diabétiques. / Atherosclerosis is closely linked to type 2 diabetes. High plasmatic concentrations of free fatty acids (FFA) and insulin are common features in patients suffering from both diseases. FFA, present in our diet, are part of the environment to which body cells are exposed. Their effects are dependent of their nature, being harmful for saturated fatty acids (SFA) and more protective for monounsaturated fatty acids (MUFA). They can have therefore various effects on vascular smooth muscle cells (VSMC) implicated throughout the development of atherosclerosis.
Thus, this study aimed to assess the impact of two FFA combinations on VSMC viability, whether or not in a hyperinsulinemic condition. Both combinations contained the same FFA but in different proportions, one being richer in SFA and the other in MUFA. Our results showed that FFA combinations have a pro-apoptotic impact, mainly due to SFA. However, the presence of oleic acid in the combinations attenuated this effect. Furthermore, oleic acid had the capacity to reduce more strongly VSMC apoptosis when combined with a SFA than when used alone, although only under specific FFA ratios. This impact is even more effective in presence of insulin.
These results highlight the presence of a competition between anti- and pro-apoptotic mechanisms dependent of FFA ratios (SFA vs. MUFA) and insulinemia to which are exposed VSMC. They also underline the importance of the presence of MUFA such as oleic acid in diets rich in SFA and could be useful for the development of new diets adapted to atherosclerotic and diabetic patients.
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Perturbation de la production de la parole chez le patient atteint d'une paralysie laryngée : données acoustiques et aérodynamiques / Perturbation of speech production in patients with laryngeal paralysis : acoustic and aerodynamic dataXiu, Noé 18 December 2018 (has links)
Notre thèse vise à étudier les conséquences d’une ablation totale ou partielle de la glande thyroïde suite à un dysfonctionnement thyroïdien, suivie ou non d’un traitement radiothérapique, et ce dans le domaine de la phonétique clinique. Ce type d’intervention perturbe généralement le système de production de la parole et conduit parfois à une dégradation de la qualité vocale de façon permanente (moins de 5% des cas) ou passagère. Le travail se veut une contribution aux recherches menées en linguistique et phonétique cliniques, plus particulièrement dans le cadre de l’analyse de faits compensatoires ou de réajustements mis en place par des patients, suite à une perturbation provoquée au niveau du fonctionnement de leur système phonatoire. Le travail a été effectué en collaboration avec le Groupe Hospitalier Saint-Vincent, et plus particulièrement avec la Clinique Sainte-Anne de Strasbourg, où se trouve le département de chirurgie thyroïdienne. Notre étude est longitudinale puisque nous avons suivi une cohorte de patients opérés de la glande thyroïde durant une année au moins, à raison d’une acquisition de données acoustiques et aérodynamiques par mois, l’examen post-opératoire ayant révélé ou non une lésion de la mobilité des plis vocaux. Nous avons étudié les possibles stratégies de compensation ou de réajustement que les patients ont pu mettre en place seuls ou à l’aide d’une rééducation orthophonique, et ce afin d’évaluer la flexibilité du système de production de la parole. Il s’agit donc d’étudier la flexibilité du système de production et de perception de la parole et de tenter de comprendre ce système à partir d’un dysfonctionnement d’origine pathologique. Il est ainsi question de déterminer les limites des déviations physiques imposées par les exigences linguistiques de clarté du système de perception de la parole. Par les différentes investigations que nous avons menées, nous avons tâché de rendre compte de la viabilité possible des unités phonétiques et phonologiques perceptivement stables, malgré une variabilité omniprésente dans le substrat physique, articulatoire, physiologique et acoustique. Une attention particulière est accordée aux dimensions sociétales liées à la qualité de vie (fatigue vocale, satisfaction des productions linguistiques, considération de soi, etc.). / Our thesis aims at studying the consequences of total or partial removal of the thyroid gland due to thyroid dysfunction, followed or not by a radiotherapy treatment, in the field of clinical phonetics. This type of intervention usually perturbs the speech production system and sometimes leads to permanent (less than 5% of cases) or transient degradation of voice quality. The work intends to be a contribution to research carried out in clinical linguistics and phonetics, more particularly in the area of compensatory or readjustment phenomena developed by patients, following perturbation provoked in their phonatory system. The work was carried out in collaboration with the Group Saint-Vincent Hospital, and more particularly with the Clinique Sainte-Anne of Strasbourg, within the department of thyroid surgery. Our study is longitudinal since we have followed a cohort of patients, who underwent thyroid gland surgery, for at least one year, acquiring acoustic and aerodynamic data every month, the postoperative examination having revealed or not a lesion in the mobility of the vocal folds. We have studied possible compensation or readjustment strategies that patients were able to deploy by themselves or with the help of speech therapy, in order to assess the flexibility of the speech production system. The purpose is thus to evaluate the flexibility of the speech production and perception system and to try to understand how this system works based on a specific dysfunction of pathological origin. It is thus a question of determining the limits of physical deviations imposed by linguistic requirements of clarity of the speech perception system. Through the various investigations that we have conducted, we have tried to account for possible viability of perceptually stable phonetic and phonological units, despite an omnipresent variability in the physical, articulatory, physiological and acoustic substrate. Particular attention is paid to societal dimensions related to quality of life (vocal fatigue, satisfaction of linguistic productions, self-esteem, etc.).
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La supervision contreproductive et ses effets sur l'efficacité des équipes de travail.McRae, Kim 08 1900 (has links)
No description available.
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Efeitos biológicos e avaliação dose-resposta das partículas de exaustão do diesel sobre o desenvolvimento embrionário inicial de camundongos / Biological effects and dose-response assessment of diesel exhaust particles on in vitro early embryo development in miceJanuário, Daniela Aparecida Nicolosi Foltran 12 March 2010 (has links)
Experimentos anteriores realizados em nosso laboratório indicam que o sucesso gestacional é afetado pela poluição atmosférica. O presente estudo teve como objetivo avaliar os efeitos biológicos associados a uma curva dose resposta das partículas de exaustão do diesel (PED) sobre o desenvolvimento embrionário inicial e o potencial de implantação, utilizando-se como modelo a fertilização in vitro e o cultivo embrionário de camundongos. No Experimento 1, encontrou-se um efeito negativo dose-dependente sobre o desenvolvimento embrionário inicial, o processo de eclosão, a alocação das células e a morfologia da massa celular interna (MCI) dos blastocistos. A análise post-hoc revelou que o desenvolvimento precoce do embrião não foi afetado pelas concentrações de 0,2 µg/cm2 ou 2 µg/cm2, mas foi significativamente afetado pela concentração de 20 µg/cm2 de PED. O processo de eclosão foi prejudicado pelas concentrações de 2 µg/cm2 e 20 µg/cm2. A alocação das células da MCI e a relação entre as células da MCI e do trofectoderma foram significativamente afetadas por todas as concentrações. Adicionalmente, observou-se um efeito negativo sobre a morfologia da MCI para as concentrações de 2 µg/cm2 e 20 µg/cm2. O Experimento 2, apesar de não mostrar efeito significativo sobre o potencial de implantação, evidenciado pela capacidade de adesão dos blastocistos e crescimento trofoblástico, revelou que a morfologia da MCI no dia 8 de cultivo, as taxas de viabilidade e de apoptose celular e a expressão de Oct4 e Cdx2 foram significativamente afetadas. O teste HSD-Tukey demonstrou que a presença de PED (0,2 µg/cm2 e 2 µg/cm2) durante o desenvolvimento embrionário aumentou significativamente a taxa de células em apoptose dos embriões tanto no dia 5 quanto no dia 8 de cultivo e, embora a proporção de células viáveis no dia 8 tenha sido prejudicada por ambas as concentrações, apenas a exposição a 2 µg/cm2 de PED diminuiu a viabilidade celular no dia 5. Por outro lado, tanto a concentração de 0,2 µg/cm2 como a de 2 µg/cm2 tiveram um efeito negativo significativo sobre a qualidade da MCI no dia 8 e a taxa de expressão de Oct4 nos blastocistos e aumentaram a porcentagem de células desses blastocistos expressando Cdx2, adicionalmente, a razão Oct4/Cdx2 dos embriões expostos a 0,2 µg/cm2 e 2 µg/cm2 foi significativamente menor. Frente a esses resultados, presumi-se que as PED poderiam estar envolvidas nos mecanismos que levariam à diminuição do sucesso reprodutivo observado em camundongos expostos à poluição atmosférica ambiental / Previous experiments conducted in our laboratory demonstrate that successful pregnancy is affected by air pollution. The aim of this study was to evaluate the biological effects associated with a dose-response curve of the diesel exhaust particles (DEP) on early embryonic development and implantation potential, using mice in vitro fertilization and culture embryo as model. In Experiment 1, we found a negative dose-dependent effect on the embryonic development, hatching process, cell allocation and morphology of inner cell mass (ICM) of blastocysts. A post-hoc analysis revealed that the early development of the embryo was not affected by concentrations of 0.2 g/cm2 or 2g/cm2, but was significantly affected by the concentration of 20 g/cm2 of DEP. The hatching process was impaired by concentrations of 2 g/cm2 and 20 g/cm2. Cell allocation of ICM and the ratio between cells of ICM and trophectoderm were significantly affected by all concentrations. Addicionaly, we observed a negative effect on ICM morphology was observed for the 2 µg/cm2 and the 20 µg/cm2 concentrations. Experiment 2, despite showing no significant effect on implantation potential, as evidenced by the adhesion ability and trophoblast outgrowth, revealed that ICM morphology on day 8 of culture, rates of cell viability and apoptosis, and expression of Oct4 and Cdx2 were significantly affected. The Tukey HSD test showed that presence of DEP (0.2 g/cm2 and 2 g/cm2) during embryonic development increased significantly the rate of apoptotic cells in embryos as on day 5 as on day 8 of culture, although the proportion of viable cells on day 8 was impaired by both concentrations, only exposure to 2 g/cm2 PED decreased cell viability on day 5. On the other hand, both the concentration of 0.2 g/cm2 such as 2 g/cm2 had a significant negative effect on the quality of ICM on the day 8 and the rate of expression of Oct4 on blastocysts, and increased the percentage of cells from these embryos expressing Cdx2, also, Oct4/Cdx2 ratio were significantly lower in the blastocysts derived from embryos exposed to 0.2 g/cm2 and 2 g/cm2¬ concentrations. Given these results, the suggestion is that DEP could be involved in the mechanisms that lead to decreased reproductive success observed in mice exposed to environmental pollution
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Avaliação in vitro da viabilidade de Enterococcus faecalis e Candida albicans nos túbulos dentinários após a aplicação de hidróxido de cálcio e clorexidina gel 2% / In vitro evauluation of the viability of Enterococcus faecalis and Candida albicans in dentinal tubules after placement of calcium hydroxide and chlorhexidine gel 2%Delgado, Ronan Jacques Rezende 12 June 2007 (has links)
Uma infecção pulpar pode resultar na colonização microbiana de todo sistema de canais radiculares incluindo os túbulos dentinários. Estes microorganismos e seus produtos tóxicos são responsáveis pelo desenvolvimento e persistência da periodontite apical de origem endodôntica. O presente estudo objetivou avaliar a viabilidade de E. faecalis e C. albicans em túbulos dentinários após a aplicação de hidróxido de cálcio, clorexidina gel 2%, hidróxido de cálcio associado à clorexidina gel 2% e soro fisiológico, através da análise por cultura microbiológica e microscopia de fluorescência. Para tanto 120 raízes de dentes humanos foram padronizadas e autoclavadas, sendo posteriormente divididas em 2 grupos (n= 60) para contaminação com E. faecalis e C. albicans por 21 dias. Em seguida, foram divididas em 8 grupos (n= 15) para aplicação das substâncias antimicrobianas nos canais radiculares e posterior incubação em estufa por 14 dias. Amostras da dentina radicular na extensão de 0 - 100 µm e de 100 - 200 µm foram coletadas e submetidas à cultura microbiológica através do plaqueamento em meios de cultura. Após 48 horas de incubação promoveu-se a avaliação das UFC. Paralelamente, as amostras foram processadas para análise em microscopia de fluorescência com auxílio de marcadores fluorescentes específicos, a fim de se determinar a proporção de microorganismos viáveis e não viáveis. Outros 6 espécimes foram preparados para análise em MEV. Os resultados mostraram uma maior capacidade de penetração intratubular para E. faecalis quando comparado a C. albicans. A aplicação de medicação intracanal resultou em significativa redução da viabilidade dos microorganismos quando comparado ao grupo controle independente da medicação aplicada e em ambas as porções da dentina radicular avaliadas. Entretanto, ao compararmos individualmente as medicações, observamos o melhor desempenho da clorexidina gel 2 % e da associação de hidróxido de cálcio e clorexidina gel 2% sem diferença significante entre elas. O hidróxido de cálcio apresentou os piores resultados para desinfecção dos canais radiculares contaminados com E. faecalis e C. albicans. Estes achados foram confirmados tanto pela cultura microbiológica quanto pela microscopia de fluorescência. A cultura microbiológica e a microscopia de fluorescência são métodos adequados e complementares para avaliação da viabilidade de E. faecalis e C. albicans e a eficácia da clorexidina gel 2% e da associação hidróxido de cálcio e clorexidina gel 2% justificam seu uso em endodontia como medicação intracanal. / A pulp infection can result in a microbial colonization of the entire root canals system, including dentinal tubules. These microorganisms and their toxic product are responsible for the development and persistence of apical periondontitis from endodontic source. The present study aimed to evaluate E. faecalis and C. albicans viability in dentinal tubules after the application of calcium hydroxide, chlorhexidine gel 2%, calcium hydroxide associated to chlorhexidine gel 2% and physiological solution, through the analysis by microbiological culture and fluorescence microscopy. For that, 120 human teeth root were standardized and submitted to autoclave, and after ere divided into 2 groups (n=60) for E. faecalis and C. albicans contamination for 21 days. Following this, they were divided into 8 groups (n=15) for application of antimicrobial substances in the root canals and subsequent incubation for 14 days. Samples from root dentin with 0 - 100 µm and 100 - 200 µm of extension were collected and submitted to microbiological culture.After 48 hours of incubation, it was performed the evaluation of colony-forming units (CFU). At the same time, the samples were processed for fluorescence microscopic analysis with the assistance of specific fluorescent markers, in order to determine the proportion of viable and non-viable microorganisms. Other 6 specimens were prepared for the analysis of scanning electron microscopy. Results demonstrated a higher capacity of penetration in the tubules for E. faecalis than for C. albicans. The application of intracanal medication resulted in significant reduction of microorganisms viability when compared to the control groups independently of the medication used and in both evaluated portions of root dentin. However, when one compares individually the medications, it was observed a better performance of chlorhexidine gel 2% and the association of calcium hydroxide with chlorhexidine gel 2% without a significant difference between them. Calcium hydroxide had the worst results for disinfection of root canal contaminated with E. faecalis and C. albicans. These findings were confirmed for the microbiological culture as well as for the fluorescence microscopy. The microbiological culture and the fluorescence microscopy are adequate methods and complementary for the evaluation of E. faecalis and C. albicans viability and the efficacy of chlorhexidine gel 2% and the association of calcium hydroxide with chlorhexidine gel 2% warrant their use in Endodontics as an intracanal medication.
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Eficiência de dietas larvais para produção massal de Ceratitis capitata e Anastrepha fraterculus (Diptera: Tephritidae) / Efficiency of larval diets for mass rearing of Ceratitis capitata and Anastrepha fraterculus (Diptera: Tephritidae)Maset, Bruno Aguiar 22 February 2019 (has links)
Várias espécies de moscas-das-frutas têm sido alvo de investigações visando sua criação massal, incluindo Ceratitis capitata (Wiedemann) (mosca-do-Mediterrâneo ou medfly) e, mais recentemente, Anastrepha fraterculus (Wiedemann) (mosca-das-frutas sul americana), sendo as espécies de maior importância econômica e quarentenária em várias áreas produtoras de frutas do Brasil, Uruguai, Argentina e Peru. Um dos fatores determinantes para que a Técnica do Inseto Estéril (TIE) seja aplicada contra uma determinada praga é a existência de métodos de criação massal da espécie. Para se alcançar a criação em larga escala dessas espécies de moscas-das-frutas, é necessário, inicialmente, o preparo de uma dieta larval adequada. Como a dieta de criação pode oferecer uma quantidade enorme de substâncias nutritivas, as larvas criadas artificialmente amadurecem mais cedo do que as selvagens, o que acelera a sucessão de gerações. Essas mudanças detectadas durante o estágio larval podem formar adultos com maiores reservas nutritivas, atingindo a maturidade sexual mais cedo. Existem várias biofábricas de mosca-das-frutas no mundo capazes de produzir machos estéreis que são utilizados para a erradicação ou supressão populacional em área-ampla. Visando-se diminuir os custos de produção dos insetos, pesquisas voltadas para a otimização e desenvolvimento de novas dietas não podem parar. Portanto, os objetivos deste trabalho foram determinar a adequação e densidade larval ideal das moscas C. capitata e A. fraterculus em novas dietas à base de farinha de milho (FM), bagaço de cana-de-açúcar (BC), pó de cenoura liofilizada (PC) e carragenina (MC), escolhendo a dieta mais eficiente e viável economicamente para produção de cada uma dessas espécies com base na quantidade e qualidade de insetos produzidos e no custo de produção. As densidades larvais ideais para a criação de C. capitata nas dietas larvais FM, BC e PC foram, respectivamente, 0,5; 2,0; e 1,0 mL de ovos/kg de dieta. A dieta larval com maior produtividade foi a dieta BC, produzindo uma média de 6.514 pupas/kg de dieta, seguida das dietas FM e PC, que produziram, respectivamente, uma média de 5.018 e 4.135 pupas por kg de dieta. A ordem decrescente de viabilidade econômica entre as três dietas larvais para C. capitata foi a mesma (BC>FM>PC), sendo necessário um investimento médio de R$ 471,29; R$ 482,26; e R$ 1.085,85, respectivamente, para a produção de 1 milhão de pupas. Os resultados com A. fraterculus mostraram que as densidades larvais ideais nas dietas FM e MC foram de 2,0/kg de dieta e 1,5 mL de ovos/L de dieta, respectivamente. A dieta larval PC foi considerada inviável devido à baixa produtividade e peso de pupas. Em termos de produtividade, a dieta larval MC (produção média de 22.451 pupas/L de dieta), foi mais eficiente do que a FM (6.688 pupas/kg de dieta). Quanto à viabilidade econômica, a dieta larval MC foi mais vantajosa, demandando um investimento médio de R$ 312,67 para a produção de 1 milhão de pupas, enquanto que a dieta FM custou R$ 1.266,36 para produzir a mesma quantidade de pupas / Several species of fruit flies have been the subject of research for their mass rearing, including Ceratitis capitata (Wiedemann) (Mediterranean fruit fly or medfly) and, more recently, Anastrepha fraterculus (Wiedemann) (South American fruit fly), being these species the ones with most economic and quarantine importance in several fruit producing areas of Brazil, Uruguay, Argentina and Peru. One of the requirements for the Sterile Insect Technique (SIT) is the existence of mass rearing methods of the target species. In order to achieve the large-scale rearing of these species of fruit flies, it is necessary initially to develop a suitable larval diet. As the rearing diet can offer a huge amount of nutrients, artificially reared larvae mature earlier than wild ones, what accelerates the succession of generations. These changes during the larval stage may form adults with higher nutritional reserves and they can reach sexual maturity earlier. There are several mass-rearing facilities for fruit flies in the world capable of producing sterile males that are used for population eradication and suppression in area-wide. In order to reduce insect production costs, the optimization or development of new diets cannot stop. Therefore, the objectives of this study were to determine the adequacy and optimal larval density of C. capitata and A. fraterculus flies in new diets based on corn flour (FM), sugarcane bagasse (BC), lyophilized carrot powder (PC) and carrageenan (MC), choosing the most efficient and less costly diet for the production of each of the species, based on the quantity and quality of insects reared and production costs. The optimal larval densities for C. capitata in larval diets FM, BC and PC were, respectively, 0.5; 2.0 and 1.0 mL of eggs/kg of diet. The larval diet that reached the highest productivity was the BC diet, giving an average of 6,514 pupae/ kg of diet, followed by FM and PC diets, which produced an average of 5,018 and 4,115 pupae per kg of diet, respectively. The decreasing order of economic viability among the three larval diets used was the same (BC>FM>PC), requiring a mean investment of R$ 471.29; R$ 482.26; and R$ 1,085.85, respectively, for the production of 1 million pupae. The results with A. fraterculus showed that the ideal larval densities in FM and MC diets were 2.0 eggs/kg of diet and 1.5 mL of eggs/L of diet, respectively. The PC larval diet was considered infeasible based on the low production and weight of pupae. In terms of productivity, the MC larval diet, which resulted in an average of 22,451 pupae produced per L of diet, was more efficient than FM, which produced a mean of 6,688 pupae per kg of diet. Considering the economic costs, the MC larval diet was also superior, requiring R$ 312,67.00 for the production of 1 million pupae, while the FM diet required R$ 1,266.36 to produce the same amount of pupae
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