Interactions between peptides and biological lipid membranes play a crucial role in many cellular processes such as in the mechanism behind Alzheimer’s disease where amyloid-beta peptide (Abeta)is thought to be a key component. The initial step of binding between a surface active peptide and its target membrane or membrane receptor can involve a non specific electrostatic association where positively charged amino acid residues and a negatively charged membrane surface interact. Here, the use of high resolution MAS NMR provides a highly sensitive and non perturbing way of studying the electrostatic potential present at lipid membrane surfaces and the changes resulting from the association of peptides. The interaction between pharmacologically relevant peptides and lipid membranes can also involve incorporation of the peptide into the membrane core and by complementing the NMR approach with differential scanning calorimetry (DSC) the hydrophobic incorporation can be studied in a non invasive way. By using 14N MAS NMR on biological lipid systems for the first time, in addition to 31P, 2H NMR and differential scanning calorimetry (DSC), gives a full picture of the changes all along the phospholipid following interactions at the membrane interface region. Being able to monitor the full length of the phospholipid enables us to differentiate between interactions related to either membrane surface association or hydrophobic core incorporation. This approach was used to establish that the interaction between nociceptin and negatively charged lipid membranes is electrostatic and hence that nociceptin can initially associate with a membrane surface before binding to its receptor. Also, it was found that Abeta can interact with phospholipid membranes via two types of interactions with fundamentally adverse effects. The results reveal that Abeta can associate with the surface of a neuronal membrane promoting accelerated aggregation of the peptide leading to neuronal apoptotic cell death. Furthermore it is also shown that Abeta can anchor itself into the membrane and suppress the neurotoxic aggregation of Abeta.

Σκοπός της παρούσας διατριβής ήταν να μελετηθεί η επίδραση του μηχανικού-ακουστικού ερεθισμού στη βιοφυσική εικόνα του εμβρύου με μια προοπτική τυχαιοποιημένη μελέτη Το υλικό της μελέτης απετέλεσαν γυναίκες με μονήρεις κυήσεις, ηλικία κύησης 30 + 0 εβδομάδες και βιοφυσική εικόνα ≤ 8/10. Οι γυναίκες με τυχαίο τρόπο κατατάσσονταν σε μια από δύο ομάδες. Στην ομάδα Α εφαρμοζόταν ακουστικός ερεθισμός διάρκειας 3 δευτερολέπτων με ένα τεχνητό λάρυγγα. Αν η βιοφυσική εικόνα παρέμενε μη φυσιολογική για 30 λεπτά, ακολουθούσε η εφαρμογή ενός δεύτερου ερεθίσματος, ίδιου με το πρώτο και εκτίμηση της βιοφυσικής εικόνας για ακόμη 30 λεπτά. Στην ομάδα Β ο χρόνος παρατήρησης παρατάθηκε για 60 λεπτά (δύο διαστήματα των 30 λεπτών) , ώστε να συμφωνεί με το χρόνο εξέτασης των γυναικών της ομάδας Α. Οι κυήσεις αντιμετωπίστηκαν με βάση την τελική βαθμολογία της βιοφυσικής εικόνας. Όσες γυναίκες γέννησαν περισσότερο από 24 ώρες από την τελευταία εξέταση δεν περιελήφθησαν στη μελέτη. Τα κριτήρια αξιολόγησης ήταν ενδομήτριος θάνατος, καισαρική τομή για εμβρυϊκή δυσπραγία, βαθμολογία Apgar < 7 στα 5 λεπτά από τον τοκετό, κεχρωσμένο αμνιακό υγρό (από μηκώνιο) και εισαγωγή στη ΜΕΘ νεογνών. Η μηδενική υπόθεση ήταν ότι η εφαρμογή του ακουστικού ερεθισμού δεν μεταβάλλει τις στατιστικές παραμέτρους της δοκιμασίας. Συνολικά 2,833 γυναίκες εισήλθαν στη μελέτη και συγκεκριμένα 1,349 στην ομάδα Α και 1,484 στην ομάδα Β. Η εφαρμογή του ακουστικού ερεθισμού μείωσε σημαντικά τον αριθμό των θετικών δοκιμασιών στην ομάδα Α σε σχέση με την ομάδα Β (4.74% vs. 6.67%, p < 0.05) και αύξησε την επίπτωση των κριτηρίων αξιολόγησης στην υποομάδα των γυναικών με θετική τελική δοκιμασία (positive likelihood ratio: 24.1-CI 95%: 11.12-52.46 vs. 7.52-CI 95%: 4.93-11.46), χωρίς να τροποποιεί το περιγεννητικό αποτέλεσμα. Επιπλέον, η ειδικότητα, η θετική προγνωστική αξία και η ακρίβεια της μεθόδου βελτιώθηκαν σημαντικά στην ομάδα Α,όπως επίσης και η αρνητική προγνωστική αξία για τους ενδομήτριους θανάτους. Συμπερασματικά, η εφαρμογή του μηχανικού-ακουστικού ερεθισμού σε περιπτώσεις που υπάρχει υποψία για εμβρυϊκή δυσπραγία, βελτιώνει την αποτελεσματικότητα της μεθόδου (βιοφυσικής εικόνας), μειώνοντας τις ψευδώς θετικές δοκιμασίες και βελτιώνοντας την ακρίβεια της μεθόδου. Θα πρέπει δε να θεωρείται ως μέσο μιας πιο ενδελεχούς εμβρυϊκής εκτίμησης σε αυτές τις περιπτώσεις. / OBJECTIVES: To verify the effect of vibroacoustic stimulation on biophysical profile score, with a prospective randomised study. STUDY DESIGN: All women with singleton pregnancy, gestational age ≥ 30 weeks, intact membranes and biophysical profile score ≤ 8/10 entered the study, after giving written consent, and were randomised to two groups. In group A, a 3-second stimulus with an artificial larynx was applied; if biophysical profile remained abnormal for 30 minutes, a second stimulus was applied, and it was assessed again. In group B the observation time was extended for 60 minutes to match the time periods of group A. Pregnancies were managed by final test score and patients delivering more than 24 hours apart from last examination were disregarded from the study. Outcome criteria were intrauterine deaths, caesarean sections for fetal distress, Apgar score < 7 at 5 minutes postpartum, meconium-stained amniotic fluid and neonatal intensive care unit admissions. Our null hypothesis was that application of vibroacoustic stimulation does not alter test’s statistical parameters. RESULTS: 1,349 patients were randomised in group A, and 1,484 in group B (2,833 in total). When comparing group A to B, application of vibroacoustic stimulation significantly decreased the number of positive tests (4.74% vs. 6.67%, p < 0.05) and increased the prevalence of outcome criteria in this subgroup (positive likelihood ratio: 24.1-CI 95%: 11.12-52.46 vs. 7.52-CI 95%: 4.93-11.46), without altering perinatal outcome. Furthermore, specificity, positive predictive value and test accuracy were significantly improved, as well as negative predictive value for intrauterine death. CONCLUSION: Vibroacoustic stimulation improves the efficiency of biophysical profile score by decreasing false positive tests and improving test accuracy and should be considered as a means of a more thorough fetal evaluation when fetal compromise is suspected.

The affect on leaf area index (LAI) and net ecosystem production (NEP) of the mountain pine beetle (Dendroctonus ponderosae) (MPB) outbreak in British Columbia affecting lodgepole pine (Pinus contorta var. latifolia) forests was examined from 1999 to 2008. The process-based carbon (C) cycle model – Boreal Ecosystem Productivity Simulator (BEPS) with remotely sensed LAI inputs was used to produce annual NEP maps, which were validated using field measurements. The annual NEP ranged from 2.43 to -8.03 MtC between 1999 and 2008, with sink to source conversion in 2000. The inter-annual variability for both LAI and NEP displayed initial decreases followed by a steadily increasing trend from 2006 to 2008 with NEP returning to near C neutrality in 2008 (-1.84 MtC). The resistance of LAI and NEP to MPB attack was attributed to ecosystem resilience in the form of secondary overstory growth and increased production of non-attacked host trees.

The affect on leaf area index (LAI) and net ecosystem production (NEP) of the mountain pine beetle (Dendroctonus ponderosae) (MPB) outbreak in British Columbia affecting lodgepole pine (Pinus contorta var. latifolia) forests was examined from 1999 to 2008. The process-based carbon (C) cycle model – Boreal Ecosystem Productivity Simulator (BEPS) with remotely sensed LAI inputs was used to produce annual NEP maps, which were validated using field measurements. The annual NEP ranged from 2.43 to -8.03 MtC between 1999 and 2008, with sink to source conversion in 2000. The inter-annual variability for both LAI and NEP displayed initial decreases followed by a steadily increasing trend from 2006 to 2008 with NEP returning to near C neutrality in 2008 (-1.84 MtC). The resistance of LAI and NEP to MPB attack was attributed to ecosystem resilience in the form of secondary overstory growth and increased production of non-attacked host trees.

Scanning ion conductance microscopy (SICM) is a technique for non-contact topographic imaging. In this thesis, a biophysical investigation into Alzheimer's Disease (AD) was carried, with toxic oligomers dosed locally and quantitatively on to single astrocytes using SICM and simultaneously monitoring the response of the target cell. Examination of the effectiveness of antibodies that bind to Abeta or alpha-synuclein (Asyn)peptides depends on the measurement of oligomer-induced abnormal calcium homeostasis in single astrocytes. The method was shown to work at physiological concentrations of oligomers. A series of experiments measuring the reduction in calcium inux in mixtures of antibodies and cerebrospinal fluid (CSF) of AD patients suggested that the binding to co-oligomers composed of Abeta and Asyn may be crucial in the treatment of AD. Furthermore, it may be beneficial to test antibodies before the clinical trial using this assay. The mechanism of this entry of calcium is hypothesised to be the result of the formation of oligomer-induced transient pores in the cell membrane. To verify this hypothesis, a new SICM instrument was built with two nanopipettes; one for dosing and one for detection of the adenosine triphosphate (ATP) release from these pores. A variety of different ATP sensors were made. The best had a sensitivity of 10 micro molar and works as a hexokinase-cofunctioned electrolyte-gated organic field-effect-transistor. However no statistically significant results for ATP release have been obtained in the experiments performed to date. Overall this thesis describes new biophysical methods to study the effect of protein aggregates on live cells and the effectiveness of potential therapies, such as antibodies and nanobodies, to reduce these aggregate induced effects. It can be applied to synthetic aggregates of Abeta or the aggregates present in human CSF.

Au cours des dernières décennies, l'Indonésie a connu des transformations spectaculaires des terres avec une expansion des plantations de palmiers à huile au détriment des forêts tropicales. L'Indonésie est actuellement l'une des régions ayant le plus haut taux de transformation de la surface terrestre dans le monde à cause de l'expansion des plantations de palmiers à huile et d'autres agricultures qui remplacent les forêts à grande échelle. Comme la végétation est un modificateur du climat près du sol, ces transformations à grande échelle ont des impacts majeurs sur les variables biophysiques de surface telles que la température de surface, l'albédo, les indices de végétation (NDVI), sur le bilan énergétique de surface et le partitionnement énergétique. Ce travail de thèse vise à quantifier les impacts des changements d’usage des terres en Indonésie sur les variables biophysiques de surface. Pour évaluer ces changements à l'échelle régionale, des données de télédétection sont nécessaires. Étant une variable clé de nombreuses fonctions écologiques, la température de surface (LST) est directement affectée par les changements de la couverture terrestre. Nous avons analysé la LST à partir de la bande thermique d'une image Landsat et produit une carte de température de surface avec une haute résolution (30m) pour les basses terres de la province de Jambi à Sumatra (Indonésie), une région qui a subi de grandes transformations au cours des dernières décennies. La comparaison des LST, albédo, NDVI et évapotranspiration (ET) entre sept différents types de couverture terrestre (forêts, zones urbaines, terres incultes, plantations de palmiers à huile jeunes et matures, plantations d'acacias et de caoutchouc) montre que les forêts ont des températures de surface inférieures à celles des autres types de couvert végétal, ce qui indique un effet de réchauffement local après la conversion des forêts vers des plantations. Les différences de LST atteignaient 10,1 ± 2,6 ºC (moyenne ± écart-type) entre les forêts et les terres déforestées. Les différences de températures de surface s'expliquent par un effet de refroidissement évaporatif des forêts, qui compense l'effet de réchauffement de l'albédo. Basé sur des différences observées dans les variables biophysiques entre les plantations de palmiers à huile jeunes et matures, nous avons analysé trois images Landsat couvrant une chronoséquence de plantations de palmiers à huile pour étudier la dynamique des variables biophysiques de surface pendant le cycle de rotation de 20-25 ans des plantations de palmiers à huile. Nos résultats montrent que les différences entre les plantations de palmiers à huile à différents stades du cycle de rotation du palmier à huile se reflètent dans les différences du bilan énergétique de surface, du partitionnement énergétique et des variables biophysiques. Au cours du cycle de rotation des plantations de palmiers à huile, les différences de température à la surface diminuent graduellement et se rapprochent de zéro autour du stade mature de la plantation de palmiers à huile de 10 ans. Parallèlement, le NDVI augmente et l'albédo diminue à proximité des valeurs typiques des forêts. Le bilan énergétique de surface et le partitionnement énergétique montrent des tendances de développement liés aux variables biophysiques et à l'âge des plantations de palmiers à huile. Les nouvelles plantations et les jeunes plantations (<5 ans) ont un rayonnement net plus faible que les plantations de palmiers à huile matures, mais ont des températures de surface plus élevées que les plantations de palmiers à huile matures. Les changements dans les variables biophysiques, le bilan énergétique et la répartition de l'énergie au cours du cycle d’une rotation du palmier à huile peuvent s'expliquer par l'effet de refroidissement évaporatif précédemment identifié dans les forêts, qui compense l'effet de réchauffement de l'albédo. L'un des principaux déterminants de ce mécanisme est la couverture végétale au cours des différentes phases du cycle de rotation du palmier à huile. Le NDVI en tant qu'indicateur du couvert végétal a montré une relation inverse cohérente avec LST de différentes plantations de palmiers à huile âgés, une tendance qui est également observée pour différents types d'utilisation des terres dans cette étude. Une analyse régionale et à plus long terme de la tendance LST entre 2000 et 2015 basée sur les données MODIS montre que dans la journée la température moyenne de Jambi a augmenté de 1,05 ºC, suivant la tendance des changements observés et dépassant les effets du réchauffement climatique. Afin d'évaluer les effets de l'expansion du palmier à huile sur le climat, le bilan énergétique de surface, le partitionnement énergétique et les processus biophysiques jouent un rôle important et le cycle complet de rotation des plantations de palmiers à huile doit être envisagé. Basé sur nos résultats, nous construisons le cycle de rotation des plantations de palmiers à huile et les changements qui se produisent au cours du développement de la végétation de palmiers à huile. Cette étude fournit des preuves que l'expansion des plantations de palmiers à huile et d'autres cultures commerciales entraîne des changements dans les variables biophysiques, réchauffant la surface du sol et augmentant ainsi l'augmentation de la température de l'air à cause du changement climatique. En utilisant des données Landsat à haute résolution, nous avons pu inclure les effets du changement d'utilisation des terres sur les variables biophysiques. Comprendre les effets du changement de la couverture terrestre sur les variables biophysiques peut soutenir des politiques concernant la conservation des forêts existantes, la planification et l'expansion des plantations de palmiers à huile et les mesures de boisement possibles. La connaissance des variables biophysiques, du bilan radiatif et de la répartition énergétique au cours du cycle de rotation du palmier à huile peut inclure de nouvelles pratiques de gestion susceptibles de réduire les conditions environnementales et microclimatiques extrêmes dans la phase initiale des plantations de palmiers à huile.

O presente trabalho descreve um programa de computador em linguagem Java que reproduz o modelo compartimental reduzido de célula mitral do bulbo olfativo construído por Davison, Feng e Brown (Brain Res. Bull. 51:393-399,2000), como uma simplificação do modelo detalhado de Bhalla e Bower (J. Neurophysiol., 69:1948-1965, 1993). O modelo reduzido considera a célula mitral como composta por quatro compartimentos, modelados conforme a metodologia de HODGKIN e HUXLEY. Por seu baixo custo computacional, o modelo reduzido permite a construção de modelos de rede de grande porte para o bulbo olfativo. A implementação computacional feita em Java apresenta grande similaridade com a original, indicando uma robustez do modelo com relação a versões em plataformas distintas. / This work describes a computer program written in Java, which reproduces the reduced compartimental model of the mitral cell of the olfactory bulb constructed by Davison, Feng and Brown (Brain Res. Bull. 51:393-399,2000), as a simplified version of the detailed model of Bhalla and Bower (J. Neurophysiol., 69:1948-1965, 1993). The reduced model considers the mitral cell as composed of four compartiments modeled according to the Hodgkin-Huxley formalism. Due to its low computational cost, the reduced model allows the construction of large-scale network models of the olfactory bulb. The computer implementation made in Java shows great similarity with the original, indicating that the model is robust with respect to implementations in different platforms.

As técnicas espectroscópicas utilizam a interação entre luz e matéria como forma de obter informações das características moleculares de um sistema. Os diversos níveis de energia manifestam-se em bandas espectrais que ao serem interpretadas fornecem características sobre estrutura, orientação e interações a nível molecular. O presente trabalho explorou as diversas facetas espectroscópicas, aliadas a técnicas biofísicas/bioquímicas, no intuito de compreender melhor dois tipos de biomoléculas importantes para a maquinaria dos seres vivos: proteínas e peptídeos. O citocromo P450 é uma enzima do tipo monooxigenase e constitui uma das maiores superfamílias de proteínas. Essa hemoproteína foi assim nomeada devido à sua característica absorção na região da Banda de Soret (450 nm), sendo esta uma particularidade natural muito utilizada em estudos por espectroscopia. Além disso, existe interesse de estudar esta família de proteínas em virtude de suas variadas funções metabólicas e biossintéticas, nos mais diversos organismos, como catalisação de esteróides, ácidos graxos, fármacos, carcinógenos químicos e metabólitos de plantas. Em especial, para a P450 de Streptomyces clavuligerus (P450CLA), ainda não se sabe ao certo como e com quais moléculas interage, e como funciona o mecanismo utilizado pela proteína para atuar em vias de síntese como a do ácido clavulânico, importante composto terapêutico. Paralelo ao paradigma de interação de uma proteína e potenciais ligantes, o entendimento dos mecanismos de interação entre peptídeos e membranas lipídicas também são de suma importância para uma melhor compreensão dos sistemas biomoleculares. Peptídeos ativos na membrana desempenham funções fundamentais no sistema de defesa de diversos organismos e decifrar os mecanismos de como essas biomoléculas agem quando inseridas em bicamadas lipídicas pode auxiliar, por exemplo, no desenvolvimento de terapias seguras e eficientes contra doenças degenerativas. Desta forma, aproveitamos as características espectroscópicas naturais de ambas as moléculas para serem empregadas em técnicas de absorção UV/vis, Dicroísmo Circular Eletrônico e Magnético, Ressonância Paramagnética Eletrônica e Ressonância Magnética Nuclear, auxiliados por técnicas termodinâmicas e de fluorescência, de modo a explorar a interação da luz com a matéria, sem interferências de sondas externas, dando enfoque às alterações de estrutura e orientação, nas mais variadas formas de interação entre moléculas de sistemas biológicos. / Spectroscopic techniques use the interaction between light and matter as a way for obtaining information about molecular characteristics of a system. The many energy levels manifest themselves in spectral bands, which when are interpreted, it provides characteristics about structure, orientation and interactions at the molecular level. The present study explored the various spectroscopic facets, allied to biophysical/biochemical techniques, in order to understand better two important biomolecules types for living beings machinery: proteins and peptides. Cytochrome P450 is a monooxygenase-like enzyme and it belongs to one of the largest proteins superfamilies. The hemoprotein received this name due its unique spectral absorption in Soret Band region (450 nm), a natural particularity widely used in spectroscopic studies. In addition, there is interest in studying this protein family by virtue of their several metabolic and biosynthetic functions in the most diverse organisms, such as steroids, fatty acids, drugs, chemical carcinogens and plant metabolites. In particular, regarding P450 from Streptomyces clavuligerus (P450CLA), it is still unclear how and which molecules it interacts with, and how the mechanism used by the protein to act in synthesis pathways such as clavulanic acid, an important therapeutic compound. Parallel to the interaction paradigm between proteins and potential ligands, the interaction mechanisms understanding between peptides and lipid membranes are also of paramount importance for a better understanding of the biomolecular systems. Active membrane peptides play key roles in the defense system of various organisms and to decipher the mechanisms how these biomolecules act when inserted into lipid bilayers, for example in the development of safe and efficient therapies against degenerative diseases. This way, we take advantage of the natural spectroscopic characteristics of both molecules to be used in UV/vis absorption techniques, Electronic and Magnetic Circular Dichroism, Electronic Paramagnetic Resonance and Nuclear Magnetic Resonance, aided by thermodynamic and fluorescence techniques, in order to explore the interaction of light with matter, without interference from external probes, focusing on changes in structure and orientation, in the most varied forms of interaction between biological systems molecules.

Submitted by Lúcia Brandão (lucia.elaine@live.com) on 2015-07-20T18:59:42Z No. of bitstreams: 1 Dissertação - Vanessa de Moura Pereira.pdf: 9133801 bytes, checksum: 2c0975213c6950b36dd18bf0eddf8511 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-22T19:57:32Z (GMT) No. of bitstreams: 1 Dissertação - Vanessa de Moura Pereira.pdf: 9133801 bytes, checksum: 2c0975213c6950b36dd18bf0eddf8511 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-22T20:12:37Z (GMT) No. of bitstreams: 1 Dissertação - Vanessa de Moura Pereira.pdf: 9133801 bytes, checksum: 2c0975213c6950b36dd18bf0eddf8511 (MD5) / Made available in DSpace on 2015-07-22T20:12:38Z (GMT). No. of bitstreams: 1 Dissertação - Vanessa de Moura Pereira.pdf: 9133801 bytes, checksum: 2c0975213c6950b36dd18bf0eddf8511 (MD5) Previous issue date: 2013-11-22 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The Amazon rainforest is home to the greatest diversity of plants and animals among all biomes on Earth. Ecosystems is of greater richness and diversity on the planet and performs environmental services of great relevance. However, changes in these ecosystems have been intensively occurring with increasing human occupation. With the creation of permanent preservation areas, the objective is to promote the protection of the natural environment of these areas that must be covered by native vegetation, contributing in this way, as a central pillar in the role of local biodiversity conservation. In this sense, this research aimed to answer the following problem: "The designation of the basin as Puraquequara Environmental Preservation Area in the Master Plan of the city of Manaus, in fact ensured the preservation of that area?". Are described biophysical impacts identified in the area as well as the possible causative agents. The results allow us to affirm that the performance of polluting activities generated large negative environmental impact on the area, leaving the competent authorities require the entrepreneur to taking immediate actions for the recovery of the affected environment, requiring the recipient to actually take the recommended action and making feasible administrative measures and appropriate legal action against those responsible. / A floresta amazônica abriga a maior diversidade de plantas e animais dentre todos os biomas da Terra. Representa os ecossistemas de maior riqueza e diversidade no planeta e realiza serviços ambientais de grande relevância. Entretanto, mudanças nesses ecossistemas vêm ocorrendo intensamente à medida que aumenta a ocupação humana. Com a criação de áreas de preservação permanentes, objetiva-se promover a proteção do ambiente natural dessas áreas que devem obrigatoriamente estar cobertas pela vegetação original, contribuindo, desta forma, como pilar central no papel de conservação da biodiversidade local. Neste sentido, esta pesquisa visou responder ao seguinte problema: “A designação da bacia do Puraquequara como Área de Preservação Ambiental no Plano diretor da cidade de Manaus, garantiu de fato a preservação da referida área?”. Encontram-se descritos os impactos biofísicos identificados na área, bem como os possíveis agentes causadores. Os resultados encontrados peritiram afirmar que a realização de atividades poluentes gerou grande impacto ambiental negativo na área, restando às autoridades competentes impor ao empreendedor a imediata tomada de ações em prol da recuperação do meio ambiente afetado, exigindo que o destinatário tome de fato as providências recomendadas e tornando exequíveis as medidas administrativas e as ações judiciais cabíveis contra os responsáveis.

Os óleos essenciais da Pimenta pseudocaryophyllus (Gomes) Landrum de planta de populações naturais de três ecossistemas, localizados na Ilha de Cananéia, região de restinga, no Morro da Cataia, cidade de Cajati, região de encosta, ambas em área de Mata Atlântica, e na Reserva Natural Morro Grande, cidade de Caldas, região de campos montanos, foram avaliados como promotores de permeação cutânea do diclofenaco de potássio. Os óleos essenciais foram extraídos de partes aéreas das plantas e o rendimento do processo foi entre 0,90% (p/p) e 2,7% (p/p). A análise da composição química mostrou diferenças, indicando tratar-se de três quimiotipos diferentes. A interação dos óleos essenciais e dos componentes majoritários com membrana biológica natural foi avaliada por FT-Raman e ATR- FTIR, indicando a interação com as porções lipídicas do tecido. Foram desenvolvidas seis membranas biológicas artificiais, compostas por ceramidas, ácidos graxos e colesterol em proporções equimolares, que foram caracterizadas por espectroscopia Raman confocal e foram consideradas semelhantes. As membranas foram utilizadas no desenvolvimento do sistema PAMPA (Parallel Artificial Membrane Permeability Assay) para avaliar a segurança e eficácia dos óleos essenciais e componentes majoritário como promotores de permeação do diclofenaco de potássio. Os resultados dos ensaios com o sistema PAMPA foram estatisticamente avaliados. A segurança foi avaliada com o critério de permeação mínima dos óleos através das membranas do sistema PAMPA, verificada pela absorbância mínima do eugenol na solução aceptora. Os óleos essenciais e componentes majoritários foram utilizados no pré-tratamento das membranas, nas concentrações de 0,125%, 0,25%, 0,50% e 2,00% (v/v) em etano!. Ensaios de permeação do diclofenaco de potássio no sistema PAMPA indicaram efeito de promoção da permeação para todos os compostos avaliados. O método de doseamento do fármaco por UV foi validado e utilizado para os ensaios de permeação de formulações de gel em base aquosa contendo o diclofenaco de potássio (1,0% p/p). As amostras de gel foram preparadas com o óleo procedente de Morro Grande, selecionado na etapa de avaliação de segurança, a 0,125% (p/v). Adicionalmente, foram preparadas formulações com citronelol e etanol, na mesma concentração. O óleo essencial da Reserva Natural Morro Grande teve efeito de promoção da permeação superior ao do citronelol e etanol, que foram equivalentes. / The essential oils of the species Pimenta pseudocaryophyllus (Gomes) Landrum collected from natural populations of three existing ecosystems in the Cananéia Island, located at sea level, Cajati city, located in hillside region, both in the Atlantic Forest areas, as well as species collected in the Morro Grande Natural Reserve, region of montane fields, were evaluated as skin permeation enhancers of potassium diclofenac. Essential oils were extracted from the aerial parts of the plants and the process yield was between of 0.90% (w/w) and 2.7% (w/w). The chemical composition analysis showed differences between the plants of three origins, indicating that they are different chemotypes. The interaction of the essential oils and their major components with natural biological membrane was evaluated by FT- Raman and ATR-FTIR, indicating interaction with the Iipid portions of the natural membrane. Six artificial biological membranes have been developed, consisting of ceramides, cholesterol and fatty acids in equimolar proportions, which were characterized by confocal Raman spectroscopy and found to be similar. The membranes were used in developing the PAMPA (Parallel Artificial Membrane Permeability Assay) system to evaluate the safety of the potential permeation enhancers. The test results with PAMPA system were statistically evaluated. Safety was evaluated with the criterion of minimum permeation of the essential oil through the membranes, checked by the minimum absorbance of eugenol in the acceptor solution. The essential oils and the major components were used in the pretreatment of the membranes, at concentrations of 0.125%, 0.25%, 0.50% and 2.00% (v/v) in ethanol. Results indicated permeation enhancement effect for ali compounds evaluated. The analytical method for the quantification of potassium diclofenac was validated and used for the evaluation of the permeation of aqueous based gel formulations containing potassium diclofenac (1.0% w/w). The gel samples were prepared with the oil from Morro Grande Natural Reserve, selected in the safety evaluation step, at 0.125% (w/v). In addition, formulations were prepared with citronellol and ethanol at the same concentration. The essential Gil of Morro Grande Natural Reserve was more efficient as permeation enhancer than citronellol and ethanol under the test conditions.