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The green python Morelia viridis is a most striking animal. Individuals are born either brick red or bright yellow and both colours change to green as adults. These colours and the remarkable colour change have long made them of interest to biologists and in demand for the pet trade. Despite this interest nothing is known of their distribution, biology or ecology in the wild. Here I address this knowledge gap by presenting results from the first detailed study of the species, at Iron Range on eastern Cape York Peninsula, Australia.¶ Individual growth was described by the von Bertalanffy growth curve, with a maximum predicted size of 1.35 metres snout-vent length. Males matured at 2.4 years and females at 3.6 years, and growth was indeterminate after approximately 12 years. The colour change from yellow to green occurs at 55 centimetres, which corresponds to individuals approximately a year old. There was no sexual dimorphism in adults, however juvenile females had larger heads than juvenile males. Adult sized individuals comprised ~50% of the population.¶ Females had a home range of 6.2 ± 1.9 ha (mean ± SE), which was positively correlated with their snout-vent length. Males adopted a roaming strategy through suitable habitat while juveniles were restricted to areas where more light reached the ground. There was overlap between multiple female home ranges, and between female home ranges and the movement paths of males. There were no differences in the distances moved by males and females of any size, although the variation in movement distances was greater in the dry season than the wet season.¶ Green pythons are obligate ambush predators which eat a variety of prey. They show an ontogenetic shift from invertebrates and terrestrial, diurnal reptiles to birds and terrestrial, nocturnal mammals. This diet change is concurrent with a shift in the time of hunting, and the location and characteristics of ambush sites. Yellow individuals were usually found within ten metres of the ground, while green individuals used the full vegetation strata and were often found in the canopy.¶ The three colour morphs of the green python appear to be adaptive for camouflage rather than intraspecific communication, as conspicuousness of each morph was always greater to a predator than to that of a conspecific. Using advanced light analysis techniques I show that each colour morph is adaptive for camouflage from visually orientated avian predators under different environmental conditions. Yellow and red morphs are half as conspicuous as green individuals would be in locations near the ground where juveniles hunt during the day. Green was the least conspicuous morph in only the canopy, where it was half as conspicuous as either the red or yellow morph. In both leafy and non-leafy sub-canopy environments green individuals were more conspicuous than both yellow and red morphs. Red morphs were least conspicuous in only the non-leafy sub-canopy environment. The conspicuousness of green males decreased with age, but this was not the case with green females. Predation of plasticine models of the three colour morphs showed that red models were ten times more likely to be predated than either green or yellow morphs, however the model colours did not always match the real morph colours.¶ There is a large predicted global distribution in Papua New Guinea, including some offshore islands, however the Australian range is restricted to small areas of eastern Cape York Peninsula. In Australia green pythons occurred in nine regional ecosystems, with most records for the closed semi-deciduous mesophyll vine forest ecosystem. A mark-recapture study at Iron Range captured 101 individuals 147 times over two wet seasons, which equates to a population size of 227 ± 81 individuals in the study area of 51 hectares. Based on the known population structure at this site only 114 (or 50%) of these individuals are adult. Although green pythons have a high density at the one intensely studied site and are predicted to occur over a large geographic area, my data are insufficient to conclude that the species is not vulnerable.

Protective coloration such as aposematism and crypsis occurs in many insects but only a few species alter their defensive strategy during the same instar. We hypothesize the adult shield bug Graphosoma lineatum with an alternating black and non-melanised longitudinal striation exhibit such a change in defensive coloration. In Sweden, the non-melanised stripes of the pre-hibernation G. lineatum are pale brown and cryptic but they change during hibernation to red and aposematic. We have tested the adaptive functions of coloration of the two G. lineatum forms against bird predators. In Paper I we used great tits as predators and measured detection time of the two forms against a background of dry grass and plants, simulating late-summer conditions. We found that the birds took longer time to find the pale than the red form. Thus, the pale form of G. lineatum is more cryptic in a dry environment than the red form. In Paper II and III we used naïve predators and measured attack rate/latency on red and pale adults and fifth-instar larvae (black and brown) to investigate avoidance and generalisation between the stages. In Paper II domestic chicks initially found the red form most intimidating, but both adult forms are more intimidating than the larva. Moreover, there was a broad generalisation among forms. In Paper III naïve great tits did not find the red form significantly more aversive than the pale adult. Neither the chicks nor the tits showed any difference in the speed of avoidance learning between the two adult colour forms. In Paper IV the shieldbugs themselves were the main focus as we compared activity levels in the different colour forms and found that G. lineatum alters behaviour in accordance to their protective strategy. Thus they were significantly less active during the cryptic phase. Taken together, these experiments suggest that the pale brown adult invests in a cryptic strategy at the cost of reduced protection from aposematism, whereas the red adult benefits from aposematism at the cost of reduced camouflage. / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Accepted.

Magister Chirurgiae Dentium - MChD / Tooth discoloration has increased the demand by patients to pursue aesthetic treatment options. Bleaching is considered a conservative approach in performing an aesthetic treatment for discolored teeth; however colour rebound and post-operative sensitivity are among the adverse effects associated with vital bleaching. In-office bleaching systems employ the use of high hydrogen peroxide concentrations. The effects of in-office bleaching agents on the degree of colour change and the gender differences in relation to bleaching outcomes are questionable. Post-operative sensitivity can be considered a bleaching side effect and the number of patients that experience it is unknown. Aim: The aim of this study was to assess the outcome of an in-office vital bleaching technique. Objectives: The objectives of this study were to evaluate the degree of colour change after vital tooth bleaching using a 35% hydrogen peroxide gel (Yotuel® Special, Biocosmetics Laboratories, Spain), using a spectrophotometer (CM-2600d Konica, Minolta) to measure the colour change at each intervention during the bleaching process, to assess the post operative sensitivity during the first week of the intervention, to evaluate the patients' perception of the colour change and to compare it to the colour change (~E) expressed in numeric values and to investigate whether gender differences exist in relation to the outcome of the bleaching procedure. Materials and Methods: The maxillary anterior teeth of 22 patients comprising of 11 males and 11 females were. bleached with a 35% hydrogen peroxide gel (Yotuel® Special, Biocosmetics Laboratories, Spain). Pre-treatment readings of the two maxillary central incisors using a spectrophotometer (CM-2600d Konica, Minolta) were obtained. Subsequent readings were obtained after scaling and polishing, before bleaching (which was used as a baseline reading), immediately after bleaching, one week and one month postoperatively. Patients were requested to complete a form regarding postoperative tooth sensitivity and their colour perception toward the bleaching treatment. Results: The quantitative effect of the bleaching material on tooth colour showed an increase in L * values and a decrease in a* and b* values, the changes were significant (p values- 0.05) except for the mean value of b* one month after bleaching which was only significant between five and ten percent.

Increased near infrared (NIR) reflection, closely resembling the red edge found in leaves, has been known in frogs for many years. Whereas previously thought of as an isolated rarity, we have shown that it is likely far more prevalent than previously believed, occurring in multiple distinct family groups and world regions. To date, there are now 26 anuran species known to demonstrate increased NIR reflectance, from 12 different genera, 4 families, and 3 ecozones. The visible/NIR reflection spectra of each individual measured was found to be characteristic of its species; whether it was wild or captive bred; and its sex. A machine learning based classification system was demonstrated as a viable method of identifying these properties from a frog's reflection spectra alone. How this reflection spectra developed from a pre-metamorphosis froglet through to adult frog was tracked, with the gradual changes to the reflection spectra of both NIR reflective and other frogs identified as being most likely dominated by the reduction in epidermal melanophores, and the increasing number of dermal iridophores. A modified consumer camera was shown to be a viable method for rapid identification of increased NIR reflection in anurans, and was used to identify that salamanders also show variation in NIR reflection between ground dwelling and leaf sitting species. The overnight colour change in Hylomantis lemur was observed, and found to occur pre-emptively of the frog's future location; with the frogs regularly transitioning from pale green ‘daytime' colouration, to the dark brown ‘night time' colouration, while still on the green leaf surface before becoming active, and undertaking the reverse transition while still active, but shortly before returning to the leaf. It seems likely that this change is for protection from silhouetting whilst active. Optical coherence tomography images were taken of several species of frog, and found to be a viable method for non-invasive investigation of anuran skin structure, with structural differences observed between the two colourations of H. lemur. It was found that the most likely cause of the increased NIR reflection in frogs is a reduction in melanin, either by absence or substitution with pterorhodin. Although the true benefit to the frog is difficult to determine, it seems likely that cryptic thermoregulation plays a key role: the maintenance of body temperature for the purpose of camouflage from animals capable of far-infrared vision. This thesis demonstrates the legitimacy of several techniques and approaches for non-invasive study of anurans, but the ultimate scope of the project is fundamentally limited by the range of frogs available. Further insight is likely to arise from increasing this scope, applying these techniques to more frogs, from more species, in more regions, and the author wishes all future researchers the greatest success in this endeavour.

Thesis (M.Dent.), Faculty of Health Sciences, University of the Witwatersrand, 2009 / This in vitro study compares tooth bleaching and consequences of tooth surface effects of Ozicure Oxygen Activator (O3, RSA) with Opalescence Quick (Ultradent, USA) tooth bleaching. One hundred and thirty six teeth (canines, incisors and premolars), which were caries free, had no surface defects and within the colour range 1M2 and 5M3. Teeth were randomly divided into the three experimental groups: Opalescence Quick, Ozicure Oxygen Activator and control. The three experimental groups received three treatments of one hour each over three consecutive days. Tooth colour was assessed using the VITAPAN 3D MASTER TOOTH GUIDE (VITA, Germany) and VITA Easyshade (VITA, Germany). A randomized block design was used to assess tooth colour change. A General Linear Models test for analysis of variance for a fractional design with significance set at P<0.05 was used. Both bleaching methods significantly lightened the teeth, visually (P<0.0001) and digitally (P= 0.013). Tooth colour change was seen after the first hour of tooth bleaching; thereafter there was little or no benefit. The tooth type played a significant role in tooth colour change (visually P=0.0416 and digitally P=0.004). The quality of the spectrophotometer may account for the different results found compared to the tooth guide. Scanning electron microscopy showed no effect of enamel loss following bleaching. Atomic force microscopy showed a 2.5 rougher enamel surface with Opalescence Quick.

Atualmente o digluconato de clorexidina (CHX) constitui o agente antimicrobiano de uso bucal mais utilizado na Odontologia devido ao seu amplo espectro de ação contra bactérias, fungos e vírus e efeito residual. No entanto, este agente apresenta efeitos colaterais quando utilizado por longo período, podendo causar coloração extrínseca nos dentes e restaurações, alteração no paladar, sensibilidade na língua e descamação das mucosas. Estes efeitos adversos conduzem à pesquisa de novas formulações. Entretanto, até o momento, nenhum agente antimicrobiano substituiu a clorexidina. Assim, o objetivo deste estudo foi desenvolver uma nova formulação antimicrobiana à base de um extrato de planta derivado do chá verde, a epigalocatequina-3-galato (EGCG), avaliar sua ação antimicrobiana contra microrganismos cariogênicos, in vitro e in vivo, e a possibilidade de alteração de cor dental, in vitro. A atividade antimicrobiana in vitro contra S. mutans, S. sobrinus e L. casei foi avaliada por meio da determinação da concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM), e, posteriormente sua concentração antimicrobiana de uso clínico foi determinada in vivo, após seu uso por crianças com alto risco e atividade da doença cárie. A concentração de microrganismos cariogênicos foi determinada na saliva das crianças, antes e após a realização do bochecho em diferentes concentrações, até se obter a máxima ação antimicrobiana. A clorexidina a 0,12% (Periogard®) e a água destilada foram utilizadas como controle positivo e negativo, respectivamente. A possível alteração de cor dental foi avaliada na coroa de dentes decíduos e permanentes humanos anteriores recém-extraídos, por meio de espectrofotometria (Easy Shade) e obtenção dos valores absolutos de L*, a* e b* (Sistema CIELab). Foi determinada a cor inicial e após a realização de 1, 5, 10 e 30 simulações de bochechos com cada solução testada durante 1 minuto. Os resultados dos ensaios de CIM e CBM in vitro mostraram que a formulação de EGCG inibiu o crescimento de todos os microrganismos cariogênicos avaliados. A CIM de EGCG contra S. mutans, S. sobrinus e para o L. casei foi obtida nas concentrações de 125, 750 e 750 &mu;g/mL, respectivamente. A CBM de EGCG contra o S. mutans, S. sobrinus e para o L. casei foi verificada nas concentrações de 250, 1000 e 1000 &mu;g/mL, respectivamente. A CBM em comum entre os 3 microrganimos foi utilizada como concentração inicial para a avaliação da atividade antimicrobiana in vivo. A partir desse valor a concentração foi sendo dobrada até atingir o valor de 4000 &mu;g/mL, que foi considerada a concentração de EGCG de uso clínico que causou redução da microbiota salivar cariogênica (86,48%) que mais se assemelhou à clorexidina (89,89%). A simulação de bochechos com a formulação de EGCG desenvolvida (4000 &mu;g/mL) causou alteração no valor absoluto das coordenadas L*, a* e b* em dentes decíduos e permanentes, de forma semelhante à causada pela solução de clorexidina. A alteração dos valores absolutos de L* e b* foram reversíveis após a realização de profilaxia dental. Com base nas metodologias e nos resultados obtidos no presente estudo pode-se concluir que a formulação desenvolvida, à base de EGCG, apresenta efeito antimicrobiano contra microrganismos cariogênicos, in vitro e in vivo, e causa alteração de cor dental reversível. / Currently the digluconate of chlorhexidine (CHX) is the antimicrobial agent for oral use most often used in dentistry due to its broad spectrum of activity against bacteria, fungi and viruses, and residual effect. However, this agent has side effects when used for long periods, and may cause extrinsic staining on teeth and restorations, change in taste, tenderness in the language and peeling of the mucous membranes. These adverse events lead to research into new formulations. However, to date, no antimicrobial agent has replaced the chlorhexidine. The objective of this study was to develop a new antimicrobial formulation based of green tea extract, epigallocatechin-3-gallate (EGCG), evaluate their antimicrobial activity against cariogenic microorganisms in vitro and in vivo, and possibility of dental color change in vitro. The in vitro antimicrobial activity against S. mutans, S. sobrinus e L. casei was evaluated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and subsequently its antimicrobial concentration was determined in clinical use, after its use for children at high risk and activity of caries. The concentration of cariogenic microorganisms was determined in the saliva of children before and after performing the mouthwash at different concentrations to obtain maximum antimicrobial activity. The chlorhexidine 0.12% (Periogard®) and distilled water were used as positive and negative controls, respectively. The possible dental color change was evaluated on the crown of deciduous and permanent teeth freshly extracted human anterior by means of spectrophotometry (Easy Shade) and obtaining the absolute values of L *, a * and b * (CIELab system). The color was determined in the beginning and after the completion of 1, 5, 10 and 30 mouthwash simulations for 1 minute of each solution tested. In vitro results of MIC and CBM tests showed that formulation EGCG inhibited the growth of all cariogenic microorganisms evaluated similarly to chlorhexidine. The MIC of EGCG against S. mutans, S. sobrinus and L. casei was observed at concentrations of 125, 750 and 750 &mu;g/ml, respectively. EGCG MBC against S. mutans, S. sobrinus and L. casei was observed at concentrations of 250, 1000 and 1000 &mu;g/ml, respectively. The common CBM between the three microbes was used as initial concentration for the evaluation of the antimicrobial activity in vivo. From this value the concentration was being folded up to the value of 4000 &mu;g/mL, which was considered the concentration of EGCG clinical use which caused reduction in salivary microbiota cariogenic (86.48%) that most resembled the chlorhexidine (89.89%). The simulation of mouthwash with the developed formulation EGCG (4000 &mu;g/ml) caused change in the absolute value of the coordinates L *, a * and b * in deciduous and permanent teeth, similar to that caused by chlorhexidine . The changes of the absolute values of L* and b* were reversible after performing dental prophylaxis. Accordance with the procedures and the results obtained in this study it can be concluded that the formulation developed, based on EGCG, has antimicrobial effect against cariogenic microorganisms in vitro and in vivo, and causes reversible change dental color.

Atualmente o digluconato de clorexidina (CHX) constitui o agente antimicrobiano de uso bucal mais utilizado na Odontologia devido ao seu amplo espectro de ação contra bactérias, fungos e vírus e efeito residual. No entanto, este agente apresenta efeitos colaterais quando utilizado por longo período, podendo causar coloração extrínseca nos dentes e restaurações, alteração no paladar, sensibilidade na língua e descamação das mucosas. Estes efeitos adversos conduzem à pesquisa de novas formulações. Entretanto, até o momento, nenhum agente antimicrobiano substituiu a clorexidina. Assim, o objetivo deste estudo foi desenvolver uma nova formulação antimicrobiana à base de um extrato de planta derivado do chá verde, a epigalocatequina-3-galato (EGCG), avaliar sua ação antimicrobiana contra microrganismos cariogênicos, in vitro e in vivo, e a possibilidade de alteração de cor dental, in vitro. A atividade antimicrobiana in vitro contra S. mutans, S. sobrinus e L. casei foi avaliada por meio da determinação da concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM), e, posteriormente sua concentração antimicrobiana de uso clínico foi determinada in vivo, após seu uso por crianças com alto risco e atividade da doença cárie. A concentração de microrganismos cariogênicos foi determinada na saliva das crianças, antes e após a realização do bochecho em diferentes concentrações, até se obter a máxima ação antimicrobiana. A clorexidina a 0,12% (Periogard®) e a água destilada foram utilizadas como controle positivo e negativo, respectivamente. A possível alteração de cor dental foi avaliada na coroa de dentes decíduos e permanentes humanos anteriores recém-extraídos, por meio de espectrofotometria (Easy Shade) e obtenção dos valores absolutos de L*, a* e b* (Sistema CIELab). Foi determinada a cor inicial e após a realização de 1, 5, 10 e 30 simulações de bochechos com cada solução testada durante 1 minuto. Os resultados dos ensaios de CIM e CBM in vitro mostraram que a formulação de EGCG inibiu o crescimento de todos os microrganismos cariogênicos avaliados. A CIM de EGCG contra S. mutans, S. sobrinus e para o L. casei foi obtida nas concentrações de 125, 750 e 750 &mu;g/mL, respectivamente. A CBM de EGCG contra o S. mutans, S. sobrinus e para o L. casei foi verificada nas concentrações de 250, 1000 e 1000 &mu;g/mL, respectivamente. A CBM em comum entre os 3 microrganimos foi utilizada como concentração inicial para a avaliação da atividade antimicrobiana in vivo. A partir desse valor a concentração foi sendo dobrada até atingir o valor de 4000 &mu;g/mL, que foi considerada a concentração de EGCG de uso clínico que causou redução da microbiota salivar cariogênica (86,48%) que mais se assemelhou à clorexidina (89,89%). A simulação de bochechos com a formulação de EGCG desenvolvida (4000 &mu;g/mL) causou alteração no valor absoluto das coordenadas L*, a* e b* em dentes decíduos e permanentes, de forma semelhante à causada pela solução de clorexidina. A alteração dos valores absolutos de L* e b* foram reversíveis após a realização de profilaxia dental. Com base nas metodologias e nos resultados obtidos no presente estudo pode-se concluir que a formulação desenvolvida, à base de EGCG, apresenta efeito antimicrobiano contra microrganismos cariogênicos, in vitro e in vivo, e causa alteração de cor dental reversível. / Currently the digluconate of chlorhexidine (CHX) is the antimicrobial agent for oral use most often used in dentistry due to its broad spectrum of activity against bacteria, fungi and viruses, and residual effect. However, this agent has side effects when used for long periods, and may cause extrinsic staining on teeth and restorations, change in taste, tenderness in the language and peeling of the mucous membranes. These adverse events lead to research into new formulations. However, to date, no antimicrobial agent has replaced the chlorhexidine. The objective of this study was to develop a new antimicrobial formulation based of green tea extract, epigallocatechin-3-gallate (EGCG), evaluate their antimicrobial activity against cariogenic microorganisms in vitro and in vivo, and possibility of dental color change in vitro. The in vitro antimicrobial activity against S. mutans, S. sobrinus e L. casei was evaluated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and subsequently its antimicrobial concentration was determined in clinical use, after its use for children at high risk and activity of caries. The concentration of cariogenic microorganisms was determined in the saliva of children before and after performing the mouthwash at different concentrations to obtain maximum antimicrobial activity. The chlorhexidine 0.12% (Periogard®) and distilled water were used as positive and negative controls, respectively. The possible dental color change was evaluated on the crown of deciduous and permanent teeth freshly extracted human anterior by means of spectrophotometry (Easy Shade) and obtaining the absolute values of L *, a * and b * (CIELab system). The color was determined in the beginning and after the completion of 1, 5, 10 and 30 mouthwash simulations for 1 minute of each solution tested. In vitro results of MIC and CBM tests showed that formulation EGCG inhibited the growth of all cariogenic microorganisms evaluated similarly to chlorhexidine. The MIC of EGCG against S. mutans, S. sobrinus and L. casei was observed at concentrations of 125, 750 and 750 &mu;g/ml, respectively. EGCG MBC against S. mutans, S. sobrinus and L. casei was observed at concentrations of 250, 1000 and 1000 &mu;g/ml, respectively. The common CBM between the three microbes was used as initial concentration for the evaluation of the antimicrobial activity in vivo. From this value the concentration was being folded up to the value of 4000 &mu;g/mL, which was considered the concentration of EGCG clinical use which caused reduction in salivary microbiota cariogenic (86.48%) that most resembled the chlorhexidine (89.89%). The simulation of mouthwash with the developed formulation EGCG (4000 &mu;g/ml) caused change in the absolute value of the coordinates L *, a * and b * in deciduous and permanent teeth, similar to that caused by chlorhexidine . The changes of the absolute values of L* and b* were reversible after performing dental prophylaxis. Accordance with the procedures and the results obtained in this study it can be concluded that the formulation developed, based on EGCG, has antimicrobial effect against cariogenic microorganisms in vitro and in vivo, and causes reversible change dental color.

O chá-verde apresenta efeitos terapêuticos na saúde geral, principalmente em função da epigalocatequina-3-galato (EGCG), componente com comprovada ação antioxidante, antiinflamatória, antimicrobiana, inativadora de endotoxina bacteriana e mineralizadora. Nesse sentido, a EGCG poderia apresentar papel promissor como medicação intracanal, durante o tratamento endodôntico de dentes portadores de necrose pulpar e lesão periapical. Assim, o objetivo deste estudo foi desenvolver um sistema de liberação tópica de EGCG, para uso no interior do sistema de canais radiculares, caracterizar suas propriedades físico-químicas bem como avaliar a possível alteração de cor dental após sua aplicação no interior dos canais radiculares de dentes decíduos e permanentes. Inicialmente, foi realizada a caracterização físico-química da EGCG por meio de espectrofotometria em Ultravioleta/Visível (UV/Vis), fluorescência e titulação potenciométrica. A seguir, foi desenvolvida uma formulação para liberação tópica prolongada de EGCG, utilizando um veículo viscoso (PEG 400). O comportamento espectral desse novo sistema de liberação de EGCG foi avaliado em função do tempo (1, 2, 3, 4, 5, 6, 24 e 27 horas) e na presença de dentes (decíduos e permanentes). Para a análise da possível alteração de cor dental, foram utilizados 20 dentes decíduos e 20 dentes permanentes, uniradiculares, recém-extraídos, provenientes do Banco de Dentes da FORP-USP. A cor da coroa dos dentes foi determinada antes (dia 0) e 21, 42 e 56 dias após a aplicação intracanal de EGCG, veiculada em água ou PEG 400, com auxílio do espectrofotômetro digital VITA Easyshade. De acordo com os resultados físico-químicos, o espectro de absorção da EGCG na região do UV/Vis apresentou um pico máximo em 274 nm e na fluorescência um pico máximo de absorção em 393 nm. A EGCG apresentou estabilidade em função do tempo e em meio ácido, tendo comportamento espectral alterado em contato com dentes decíduos e permanentes. Em meio básico o espectro de absorção da EGCG em luz UV/Vis foi alterado para 323 nm. O sistema de liberação tópica intracanal de EGCG não alterou suas propriedades físico-químicas, mantendo os valores de absorção em UV/Vis (278 nm) e fluorescência (377 nm), sua estabilidade em função do tempo e não alterou seu comportamento espectral em contato com dentes decíduos e permanentes. A aplicação do sistema proposto de liberação tópica da EGCG, no interior dos canais radiculares de dentes decíduos e permanentes, não causou alteração de cor dental significativa, após o período de 21, 42 e 56 dias. Com base nas metodologias e nos resultados obtidos no presente estudo pode-se concluir que a EGCG apresenta fluorescência e estabilidade em função do tempo e em meio ácido, enquanto em meio básico e na presença de elementos dentais seu comportamento espectral sofre alteração. O sistema desenvolvido para liberação controlada da EGCG manteve suas propriedades físico-químicas, apresentou estabilidade em função do tempo e não foi alterou seu comportamento espectral na presença dos elementos dentais. Ainda, a aplicação intracanal deste sistema não causou alteração de cor da coroa dental. / Green tea has therapeutic effects on general health, mainly due to epigallocatechin-3-gallate (EGCG), a component with proven antioxidant, anti-inflammatory, antimicrobial, inactivating bacterial endotoxin and mineralization action. In this sense, EGCG could present promising role as an intracanal medication during endodontic treatment of teeth presenting pulp necrosis and periapical lesion. Thus, the objective of this study was to develop a system for topical delivery of EGCG, for use inside the root canal system, characterize their physical and chemical properties as well as to evaluate the possible color change after its application in root canals of deciduous and permanent teeth. Initially, was performed physicochemical characterization of EGCG by spectrophotometry Ultraviolet/Visible (UV/Vis), fluorescence, and potentiometric titulation. Next, a formulation for topical extended release of EGCG using a viscous vehicle (PEG 400) was developed. The spectral characteristic of this new delivery system of EGCG was assessed in function of the medium (acidic, basic or neutral), the time (1, 2, 3, 4, 5, 6, 24 and 27 hours) and in the presence of teeth (deciduous and permanent). To the analysis of possible tooth color changes, 20 primary teeth and 20 permanent teeth, single-rooted, freshly extracted, from the Bank of Teeth FORP-USP, were used. The color of the tooth crown was determined before (day 0) and 21, 42 and 56 days after intracanal application of EGCG diluted in water or PEG 400, using a digital VITA Easyshade spectrophotometer. According to results the physicochemical spectrum of absorption of the EGCG showed maximum absorption peak at 274 nm in the UV/Vis and at 393 nm in the fluorescence. The EGCG presented stability as a function of time and in acid medium, and its spectral behavior was changed in contact with deciduous and permanent teeth. In basic medium the EGCG absorption spectrum in UV/Vis was changed to 323 nm. The topical delivery system of EGCG intracanal did not change its physicochemical properties, maintaining the values of absorption in UV/Vis (278 nm) and fluorescence (377 nm), its stability as a function of time, and does not change its spectral contact with deciduous and permanent teeth. The application of the proposed system for topical delivery of EGCG in the root canals of deciduous and permanent teeth, did not cause significant tooth color change after the period of 21, 42 and 56 days. Based on the methodologies and the results obtained in this study it can be concluded that EGCG presents fluorescence and stability as a function of time and in an acidic medium, whereas in basic medium and in the presence of dental elements its spectral behavior is changed. The system developed for EGCG controlled release maintains its physicochemical properties, presents stability with time and does not change its spectral behavior in the presence of dental elements. Yet, the intracanal application of this system did not change the color of the teeth\'s crown.

This thesis addresses the mechanisms regulating the evolution and maintenance of colour polymorphism in the shrimp Hippolyte obliquimanus. It includes four chapters that illustrate, in an integrative approach, the ecological and behavioural consequences of different camouflage and habitat use strategies selected in main colour morphs. In the first chapter entitled Camouflage through colour change: mechanisms, adaptive value, and ecological significance, the physiological and structural mechanisms underpinning colour change, and hence camouflage, in different species were detailed within a general framework. Also in this section, both the adaptive value and the ecological and evolutionary implications of these processes were discussed. In the second chapter entitled Shape, colour plasticity, and habitat use indicate morph-specific camouflage strategies in a marine shrimp morph-specific colour change and habitat selection were investigated as potential mechanisms affecting camouflage efficiency and the natural distribution of H. obliquimanus colour morphs between macroalgal habitats formed by Sargassum furcatum and Galaxaura marginata. Moreover, morph-specific morphology and behavioural traits were also assessed. Shrimp showing a homogeneous coloration (H) changed colour when placed in colour-mismatching backgrounds and also exhibited a more robust shape associated to high substrate fidelity and specialization. On the other hand, striped translucent shrimp (ST) were not capable of colour change and showed high mobility, consistent with a more streamlined shape and a generalised use of algal habitats. In the third chapter entitled The adaptive value of camouflage and colour change in a polymorphic shrimp visual models and predation trials were used to test the adaptive value of camouflage and colour change of H. obliquimanus colour morphs against colour-contrasting macroalgae, which provide different levels of colour concealment. The low colour discrimination predicted by a visual model of a seahorse predator for pink shrimp against the red weed Galaxaura was directly translated to lower detection and consumption rates on this morph by captive seahorses in a laboratory experiment, demonstrating the high adaptive value of camouflage in this species. Finally, in the fourth chapter entitled Habitat-dependent niche partitioning between colour morphs of the algal-dwelling shrimp Hippolyte obliquimanus the trophic ecology of H and ST morphs when using Sargassum and Galaxaura habitats were assessed through stable isotope analysis. Morphs showed different isotopic signatures only in Galaxaura, where the supply of food resources are limiting, leading to a tropic niche diversification between H and ST shrimp and a probable relaxation of intra-specific competition. Conversely, in Sargassum, where resources are abundant, morphs coexistence is possible even without niche partitioning. Altogether, the results of this thesis contributed to a better understanding of the adaptive value and the underlying mechanisms controlling for the maintenance of colour polymorphism in H. obliquimanus. From both an ecological and evolutionary perspective, morph-specific strategies of camouflage and habitat use likely promote the stability of shrimp populations in a spatially heterogeneous and markedly seasonal habitat. / Esta tese aborda os mecanismos que regulam a evolução e manutenção do polimorfismo de cor no camarão Hippolyte obliquimanus ao longo de quatro capítulos que ilustram de forma integrada as consequências ecológicas e comportamentais das distintas estratégias de camuflagem e de uso do habitat selecionadas nos principais morfótipos da espécie. No Capítulo 1, intitulado Camouflage through colour change: mechanisms, adaptive value, and ecological significance, são detalhados de maneira geral os mecanismos fisiológicos e estruturais responsáveis pelo processo de mudança de cor e obtenção de camuflagem em diferentes espécies de animais. Nessa seção também é detalhado o valor adaptativo e as implicações ecológicas e evolutivas desses processos. No Capítulo 2, intitulado Shape, colour plasticity, and habitat use indicate morph-specific camouflage strategies in a marine shrimp, são investigados os processos de mudança de cor e de seleção de habitat como possíveis mecanismos reguladores dos padrões de camuflagem e de distribuição dos morfótipos de cor de H. obliquimanus entre os bancos das algas Sargassum furcatum e Galaxaura marginata. Além disso, também são avaliadas diferenças morfológicas e comportamentais entre os morfótipos na utilização dos habitats vegetados. Camarões caracterizados por uma coloração homogênea (H) mudam de cor quando em contato com substratos de coloração diferente à sua, exibindo também uma morfologia mais robusta e um comportamento de alta fidelidade e especialização aos substratos de macroalgas. Por outro lado, camarões transparentes listrados (TL) não mudam de cor e apresentam elevada mobilidade, consistente com uma morfologia mais hidrodinâmica e com uso mais generalizado dos habitats estudados. No Capítulo 3, intitulado The adaptive value of camouflage and colour change in a polymorphic shrimp, são utilizados modelos visuais e experimentos de predação para testar o valor adaptativo da camuflagem e mudança de cor em diferentes morfótipos do camarão H. obliquimanus quando estes estão associados a macroalgas que fornecem diferentes graus de semelhança cromática. O baixo grau de discriminação cromática previsto para camarões rosa em associação à alga vermelha Galaxaura através da modelagem visual de um predador (cavalo-marinho) resultou em baixas taxas de detecção e predação desse morfótipo por cavalos-marinhos em um experimento de laboratório, demonstrando um alto valor adaptativo da camuflagem nessa espécie. Por fim, no Capítulo 4, intitulado Habitat-dependent niche partitioning between colour morphs of the algal-dwelling shrimp Hippolyte obliquimanus, a ecologia trófica dos morfótipos de cor H e TL na utilização dos bancos de Sargassum e Galaxaura é avaliada através de análises de isótopos estáveis. Os morfótipos de cor mostraram diferenças nas razões isotópicas apenas em Galaxaura, onde a abundância de recursos alimentares é limitante, levando à diversificação de nicho trófico entre camarões H e TL e uma provável redução da competição intraespecífica. Por outro lado, em Sargassum, onde há abundância de recursos, a coexistência entre os morfótipos é possível mesmo sem partição de nicho ecológico. Em conjunto, os resultados obtidos nessa tese detalham tópicos centrais sobre os mecanismos reguladores e o valor adaptativo do polimorfismo cromático em H. obliquimanus. Dentro de uma abordagem ecológica e evolutiva, as diferentes estratégias de camuflagem e de uso de habitat dos morfótipos de cor parecem ser essenciais para o equilíbrio populacional da espécie, uma vez que os bancos de macroalgas utilizados como habitat pelos indivíduos são bastante heterogêneos e sua disponibilidade varia sazonalmente.