Estudo da formação de domínios em membranas modelos induzidos por peptídeos antimicrobianos e sua ação interfacial

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Pressure Dependence of N-15 Chemical Shifts in Model Peptides Ac-Gly-Gly-X-Ala-NH2

High pressure NMR spectroscopy has developed into an important tool for studying conformational equilibria of proteins in solution. We have studied the amide proton and nitrogen chemical shifts of the 20 canonical amino acids X in the random-coil model peptide Ac-Gly-Gly-X-Ala-NH2, in a pressure range from 0.1 to 200 MPa, at a proton resonance frequency of 800 MHz. The obtained data allowed the determination of first and second order pressure coefficients with high accuracy at 283 K and pH 6.7. The mean first and second order pressure coefficients <B-1(15N)> and <B-2(15N)> for nitrogen are 2.91 ppm/GPa and -2.32 ppm/GPa(2), respectively. The corresponding values <B-1(1H)> and <B-2(1H)> for the amide protons are 0.52 ppm/GPa and -0.41 ppm/GPa(2). Residual dependent (1)J(1H15N)-coupling constants are shown.

Pollen morphology of the genus Eremanthus Less. (Vernonieae, Asteraceae)

(Pollen morphology of the genus Eremanthus Less. (Vernonieae, Asteraceae)). In order to evaluate the significance of the pollen morphology for generic and infrageneric taxonomy of the genus Eremanthus (Vernonieae, Asteraceae), and to provide additional data for its phylogenetic reconstruction, the pollen of 20 of the 23 species of the genus was examined using light and scanning electron microscopy. Acetolysed pollen grains were measured, described, and illustrated using light microscopy, while non-acetolysed pollen grains were analyzed using scanning electron microscopy. Pollen grains of these species are isopolar, oblate-spheroidal in most of the species, more rarely prolate spheroidal or suboblate, subtriangular amb, tricolporate and subechinolophate. The variation among quantitative characters does not correlate with the macromorphological subdivision of the genus or with the generic or specific limits.

The influence of both urea and 2,2,2

The influence of external agents on proteins function and structure is essential to elucidate the unfolding pathways and self-assemble properties. The knowledge of the protein amyloid fibril formation process is important due to the fields that this subjected is related, in particular for the neurodegenerative disorders. In the present work we studied the influence of both urea and 2,2,2-Trifluoroethanol (TFE) and temperature on the structure and proteinprotein interactions of Bovine Serum Albumin (BSA), by means of UV-Vis spectroscopy, static fluorescence and small angle X-ray scattering technique. The experiments were performed in samples composed by 10 and 3 mg/ml of BSA at pH 5.8, near the protein pI. First, Thioflavin-T fluorescence measurements indicated that urea, in the absence of TFE, was able to increase the amyloid fibril formation of BSA at 45oC and increasing the urea concentration the rate of amyloid fibril formation also increases. Concerning the presence of TFE, SAXS data suggest that BSA tridimensional structure is not altered by the presence of TFE 5% and 10% v/v in all studied protein concentrations. Interestingly, the presence of TFE on the urea-containing BSA also increases the rate of amyloid fibril formation, as compared to the TFE-free system, indicating that TFE can catalyze the amyloid-fibril formation. The presence of TFE 20% v/v, however, induces the formation of aggregates, but at this time we were not able to infer if such aggregates are amyloidlike or amorphous. Taking together, the results give support to infer that BSA can for fibrils in the presence of urea at 45oC and TFE can act as a stabilizer or as a denaturant agent for BSA.

Numeric reconstruction of cytoskeleton with finite element method and topology optimization method

The importance of mechanical aspects related to cell activity and its environment is becoming more evident due to their influence in stem cell differentiation and in the development of diseases such as atherosclerosis. The mechanical tension homeostasis is related to normal tissue behavior and its lack may be related to the formation of cancer, which shows a higher mechanical tension. Due to the complexity of cellular activity, the application of simplified models may elucidate which factors are really essential and which have a marginal effect. The development of a systematic method to reconstruct the elements involved in the perception of mechanical aspects by the cell may accelerate substantially the validation of these models. This work proposes the development of a routine capable of reconstructing the topology of focal adhesions and the actomyosin portion of the cytoskeleton from the displacement field generated by the cell on a flexible substrate. Another way to think of this problem is to develop an algorithm to reconstruct the forces applied by the cell from the measurements of the substrate displacement, which would be characterized as an inverse problem. For these kind of problems, the Topology Optimization Method (TOM) is suitable to find a solution. TOM is consisted of an iterative application of an optimization method and an analysis method to obtain an optimal distribution of material in a fixed domain. One way to experimentally obtain the substrate displacement is through Traction Force Microscopy (TFM), which also provides the forces applied by the cell. Along with systematically generating the distributions of focal adhesion and actin-myosin for the validation of simplified models, the algorithm also represents a complementary and more phenomenological approach to TFM. As a first approximation, actin fibers and flexible substrate are represented through two-dimensional linear Finite Element Method. Actin contraction is modeled as an initial stress of the FEM elements. Focal adhesions connecting actin and substrate are represented by springs. The algorithm was applied to data obtained from experiments regarding cytoskeletal prestress and micropatterning, comparing the numerical results to the experimental ones

Quantification of cellular action fibers alignment from confocal microscopic images.

The cellular rheology has recently undergone a rapid development with particular attention to the cytoskeleton mechanical properties and its main components - actin filaments, intermediate filaments, microtubules and crosslinked proteins. However it is not clear what are the cellular structural changes that directly affect the cell mechanical properties. Thus, in this work, we aimed to quantify the structural rearrangement of these fibers that may emerge in changes in the cell mechanics. We created an image analysis platform to study smooth muscle cells from different arteries: aorta, mammary, renal, carotid and coronary and processed respectively 31, 29, 31, 30 and 35 cell image obtained by confocal microscopy. The platform was developed in Matlab (MathWorks) and it uses the Sobel operator to determine the actin fiber image orientation of the cell, labeled with phalloidin. The Sobel operator is used as a filter capable of calculating the pixel brightness gradient, point to point, in the image. The operator uses vertical and horizontal convolution kernels to calculate the magnitude and the angle of the pixel intensity gradient. The image analysis followed the sequence: (1) opens a given cells image set to be processed; (2) sets a fix threshold to eliminate noise, based on Otsu's method; (3) detect the fiber edges in the image using the Sobel operator; and (4) quantify the actin fiber orientation. Our first result is the probability distribution II(Δθ) to find a given fiber angle deviation (Δθ) from the main cell fiber orientation θ0. The II(Δθ) follows an exponential decay II(Δθ) = Aexp(-αΔθ) regarding to its θ0. We defined and determined a misalignment index α of the fibers of each artery kind: coronary αCo = (1.72 ‘+ or =’ 0.36)rad POT -1; renal αRe = (1.43 + or - 0.64)rad POT -1; aorta αAo = (1.42 + or - 0.43)rad POT -1; mammary αMa = (1.12 + or - 0.50)rad POT -1; and carotid αCa = (1.01 + or - 0.39)rad POT -1. The α of coronary and carotid are statistically different (p < 0.05) among all analyzed cells. We discussed our results correlating the misalignment index data with the experimental cell mechanical properties obtained by using Optical Magnetic Twisting Cytometry with the same group of cells.

Rotational diffusion membrane and fluorescence anisotropy.

Structural properties of model membranes, such as lipid vesicles, may be investigated through the addition of fluorescent probes. After incorporation, the fluorescent molecules are excited with linearly polarized light and the fluorescence emission is depolarized due to translational as well as rotational diffusion during the lifetime of the excited state. The monitoring of emitted light is undertaken through the technique of time-resolved fluorescence: the intensity of the emitted light informs on fluorescence decay times, and the decay of the components of the emitted light yield rotational correlation times which inform on the fluidity of the medium. The fluorescent molecule DPH, of uniaxial symmetry, is rather hydrophobic and has collinear transition and emission moments. It has been used frequently as a probe for the monitoring of the fluidity of the lipid bilayer along the phase transition of the chains. The interpretation of experimental data requires models for localization of fluorescent molecules as well as for possible restrictions on their movement. In this study, we develop calculations for two models for uniaxial diffusion of fluorescent molecules, such as DPH, suggested in several articles in the literature. A zeroth order test model consists of a free randomly rotating dipole in a homogeneous solution, and serves as the basis for the study of the diffusion of models in anisotropic media. In the second model, we consider random rotations of emitting dipoles distributed within cones with their axes perpendicular to the vesicle spherical geometry. In the third model, the dipole rotates in the plane of the of bilayer spherical geometry, within a movement that might occur between the monolayers forming the bilayer. For each of the models analysed, two methods are used by us in order to analyse the rotational diffusion: (I) solution of the corresponding rotational diffusion equation for a single molecule, taking into account the boundary conditions imposed by the models, for the probability of the fluorescent molecule to be found with a given configuration at time t. Considering the distribution of molecules in the geometry proposed, we obtain the analytical expression for the fluorescence anisotropy, except for the cone geometry, for which the solution is obtained numerically; (II) numerical simulations of a restricted rotational random walk in the two geometries corresponding to the two models. The latter method may be very useful in the cases of low-symmetry geometries or of composed geometries.

Mechanical alterations in airway smooth muscle after interaction with indoor pollutant - A mathematical model

The viscoelasticity of mammalian lung is determined by the mechanical properties and structural regulation of the airway smooth muscle (ASM). The exposure to polluted air may deteriorate these properties with harmful consequences to individual health. Formaldehyde (FA) is an important indoor pollutant found among volatile organic compounds. This pollutant permeates through the smooth muscle tissue forming covalent bonds between proteins in the extracellular matrix and intracellular protein structure changing mechanical properties of ASM and inducing asthma symptoms, such as airway hyperresponsiveness, even at low concentrations. In the experimental scenario, the mechanical effect of FA is the stiffening of the tissue, but the mechanism behind this effect is not fully w1derstood. Thus, the aim of this study is to reproduce the mechanical behavior of the ASM, such as contraction and stretching, under FA action or not. For this, it was created a two-dimensional viscoelastic network model based on Voronoi tessellation solved using Runge-Kutta method of fourth order. The equilibrium configuration was reached when the forces in different parts of the network were equal. This model simulates the mechanical behavior of ASM through of a network of dashpots and springs. This dashpot-spring mechanical coupling mimics the composition of the actomyosin machinery of ASM through the contraction of springs to a minimum length. We hypothesized that formation of covalent bonds, due to the FA action, can be represented in the model by a simple change in the elastic constant of the springs, while the action of methacholinc (MCh) reduce the equilibrium length of the spring. A sigmoid curve of tension as a function of MCh doses was obtained, showing increased tension when the muscle strip was exposed to FA. Our simulations suggest that FA, at a concentration of 0.1 ppm, can affect the elastic properties of the smooth muscle fibers by a factor of 120%. We also analyze the dynamic mechanical properties, observing the viscous and elastic behavior of the network. Finally, the proposed model, although simple, ir1corporates the phenomenology of both MCh and FA and reproduces experirnental results observed with ir1 vitro exposure of smooth muscle to .FA. Thus, this new mechanical approach incorporates several well know features of the contractile system of the cells ir1 a tissue level model. The model can also be used in different biological scales.

Influence of chest wall on lung mechanics during inspiration.

RATIONALE: The interaction between lungs and chest wall influences lung volume, that determines lung history during respiration cycle. In this study, the influence of chest wall mechanics on respiratory system is assessed by the evaluation of inspiration pressure-volume curve (PV curve) under three different situations: closed-chest, open-chest and isolated lung. The PV curve parameters in each situation allow us to further understand the role played by different chest wall elements in the respiratory function. Methods: Twenty-four male Wistar rats (236 ± 29 g) were used. The animals were weighted and then anesthetized with xylazine 2% (0,5mL/kg) and ketamine 10% (0,9mL/kg), exsanguinated and later tracheostomized with a metallic cannula (14 gauge). The cannula was connected to an automatic small animal insufflator. This setup was connected to a pressure transducer (32 samples/s). The 24 animals were randomly separated in three groups: (i) closed chest, (ii) open chest and (iii) isolated lung. The rats were insufflated with 20mL quasi-statically (constant speed of 0,1mL/s). Insufflated volume and measured pressure data were kept and PV curves were obtained for all animals. The PV curves were fitted (non-linear least squares) against the sigmoid equation (1) to obtain the sigmoid equation parameters (a,b,c,d). Elastance measurements were obtained from linear regression of pressure/volume measurements in a 0,8s interval before and after the calculated point. Results: The parameters a, b and c showed no significant change, but the parameter d showed a significant variation among the three groups. The initial elastance also varied between open and closed chest, indicating the need of a higher pressure for the lung expansion, as can be seen in Table 1. Table 1: Mean and Standard Deviation of parameters obtained for each protocol. Protocol: Closed Chest – a (mL) -0.35±0.33; b (mL) 13.93±0.89; c (cm H2O) 21.28±2.37; d (cm H2O) 6.17±0.84; r²** (%) 99.4±0.14; Initial Elastance* (cm H2)/mL) 12.72±6.66; Weight (g) 232.33±5.72. Open Chest - a (mL) 0.01±0.28; b (mL) 14.79±0.54; c (cm H2O) 19.47±1.41; d (cm H2O) 3.50±0.28; r²** (%) 98.8±0.34; Initial Elastance* (cm H2)/mL) 28.68±2.36; Weight (g) 217.33±7.97. Isolated Lung - a (mL) -0.09±0.46; b (mL) 14.22±0.75; c (cm H2O) 21.76±1.43; d (cm H2O) 4.24±0.50; r²** (%) 98.9±0.19; Initial Elastance* (cm H2)/mL) 7.13±8.85; Weight (g) 224.33±16.66. * Elastance measures in the 0-0,1 mL range. ** Goodness of sigmoid fit versus measured data Conclusion: A supporting effect of the chest wall was observed at the initial moments of inspiration, observed as a higher initial elastance in open chest situations than in closed chest situations (p=0,00001). The similar initial elastance for the isolated lung and closed chest may be explained by the specific method used for the isolated lung experiment. As the isolated lung is supported by the trachea vertically, the weight of the tissue may have a similar effect of the residual negative pressure in the thorax, responsible for maintaining the residual volume.

Speckle patterns during the spreading of lung surfactant.

Pulmonary surfactant is a very important product in the medical treatment of the syndrome of insufficiency respiratory in neonates. The synthesis of this surfactant in labs need to optimize the rate of spreading in the alveolar interstitial liquid obtaining a monolayer of the phospholipids membrane base capable to maintains several of the dynamical properties of the respiratory system during breathing. The recover of theses mechanical properties has to be archived using the minimal quantities of product and with the optimal proteins composition (SP-B in special). In this paper we show our results of obtaining and process speckle pattern images of the spreading of phospholipids membrane composed the matrix of this product (DPPC) when physiologic interstitial liquid are presented.

Utilization of fringe projection technique for evaluation of wound dimensions and of healing progress.

Background: The methods used for evaluating wound dimensions, especially the chronic ones, are invasive and inaccurate. The fringe projection technique with phase shift is a non-invasive, accurate and low-cost optical method. Objective: The aim is to validate the technique through the determination of dimensions of objects of known topography and with different geometries and colors to simulate the wounds and tones of skin color. Taking into account the influence of skin wound optical factors, the technique will be used to evaluate actual patients’ wound dimensions and to study its limitations in this application. Methods: Four sinusoidal fringe patterns, displaced ¼ of period each, were projected onto the objects surface. The object dimensions were obtained from the unwrapped phase map through the observation of the fringe deformations caused by the object topography and using phase shift analysis. An object with simple geometry was used for dimensional calibration and the topographic dimensions of the others were determined from it. After observing the compatibility with the data and validating the method, it was used for measuring the dimensions of real patients’ wounds. Results and Conclusions: The discrepancies between actual topography and dimensions determined with Fringe Projection Technique and for the known object were lower than 0.50 cm. The method was successful in obtaining the topography of real patient’s wounds. Objects and wounds with sharp topographies or causing shadow or reflection are difficult to be evaluated with this technique.

Oligonucleotide Adsorption Affects Phase Transition but Not Interdigitation of diC14-Amidine Bilayers

Oligonucleotides have been extensively used in basic research of gene expression and function, vaccine design, and allergy and cancer therapy. Several oligonucleotide-based formulations have reached the clinical trial phase and one is already on the market. All these applications, however, are dependent on suitable carriers that protect oligonucleotides against degradation and improve their capture by target cells. The cationic lipid diC14-amidine efficiently delivers nucleic acids to mammalian cells. It was recently shown that diC14-amidine bilayers present an interdigitated phase which strongly correlates with a potent fusogenic activity at low temperatures. Interdigitated phases correspond to very ordered gel phases where the two bilayer leaflets are merged; they usually result from perturbations at the interfacial region such as modifications of the polar headgroup area or dehydration of the bilayer. Interdigitation has been described for asymmetric lipids or mixed-chain lipids of different chain lengths and for lipids with large effective headgroup sizes. It has also been described for symmetric lipids under pressure modifications or in the presence of alcohol, glycerol, acetonitrile, polymyxin B, or ions like thiocyanate. Surprisingly, the role of polyelectrolytes on membrane interdigitation has been only poorly investigated. In the present work, we use dynamic light scattering (DLS), differential scanning calorimetry (DSC), and electron spin resonance (ESR) to explore the effect of a small single-stranded oligonucleotide (ODN) polyelectrolyte on the structure and colloid stability of interdigitated diC14-amidine membranes.

Conocimiento de la mecánica corporal

El personal de enfermería, es quien se ocupa de atender a los pacientes brindándoles ayuda, comodidad y apoyo. Muchas de las actividades cotidianas que realiza enfermería incluyen movimientos repetidos, traslado de pacientes, cambios de posturas y levantamiento de objetos pesados, lo que exige un considerable gasto de energía. El conocimiento y la adecuada aplicación de la mecánica corporal son un buen mantenimiento preventivo de la salud corporal, así mismo, contribuye a disminuir el riesgo de trastornos músculo-esqueléticas. Estos son un conjunto de lesiones inflamatorias y degenerativas de músculos, nervios, tendones y articulaciones. En el personal de enfermería son muy frecuentes, lo que repercute en la calidad de vida del individuo y provoca un descenso en la calidad asistencial. El objetivo principal de esta investigación es determinar la relación entre el conocimiento de la mecánica corporal y los trastornos músculo-esqueléticos, del personal de enfermería del Área de internación del Hospital Privado Quirúrgico de Mendoza, en septiembre de 2013.

Definición de un Modelo Edafogeomorfológico para la Identificación de Necesidades de Manejo y Recuperación de Suelos a Nivel Local. Caso Cuenca Cañada La Gorda, Machiques Colón, Estado Zulia, Venezuela.

El objetivo de este trabajo es generar un modelo Edafogeomorfológico útil en la identificación de necesidades de manejo de suelos, se realizó un estudio en el área de la cuenca Cañada La Gorda Machiques-Colón, estado Zulia, Venezuela, caracterizada por un clima tropical de condición subhúmeda, con duración del periodo de crecimiento (DPC) de 230 días, régimen de humedad Ustic y de temperatura Isohipertémico. Se empleó el enfoque de la ecuación factorial de formación de suelos para el análisis y descripción biofísica de los factores a lo largo de una carena. El relieve caracterizado a partir de fotografías aéreas, imágenes de satélites y de chequeos sistemático mediante transectos en el sentido del flujo del escurrimiento; la vegetación a través del uso de la tierra, la cobertura vegetal, la identificación de las especies dominantes a partir de sus nombres vernáculos y la definición de indicadores de vegetales (Iv). Los suelos fueron descritos y clasificados según la Taxonomía de suelos y valorados mediante el modelo paramétricode Riquier et al. (1970) para determinar el índice de productividad (Ip). Se caracterizaron dos paisajes gemorfológicos: Colinar (C) y Valle (V), seis posiciones geomorfológicas entre ambos paisajes definidas por la sucesión de relieves en el sentido de la pendiente: Tope de colina-loma (TC), mesa conservada (MC), vertientes de mesa alta (VA), media (VM), baja (VB) y valle intracolinar (VI); e igual número de perfiles de suelos representativos, los cuales mostraron edafogénesis muy avanzada con Ip inferiores a 8% en todas las posiciones, exceptuando la VB, con una productividad de 13%. El uso de la tierra es a base de pastoreo semi-intensivo de plantas forrajeras introducidas. Las formaciones vegetales predominante fueron los matorrales y arbustales dispersos, acompañados con restos de una selva tropófila fuertemente afectada por la extracción forestal y la conversión en áreas de pastoreo. Se identificaron 8 Iv, asociados fuertemente con condiciones de físicas e hidrológicas del suelo. El alto impacto de las actividades humanas sobre el suelo y vegetación, expresado a través de los procesos de erosión activa, la ausencia de áreas boscosas y la baja productividad de los sistemas de ganadería reportada para la zona, señalan la necesidad de reorientación del uso actual de la tierra, para lo cual se plantean alternativas como la incorporación de bosques protectores y sistemas agrosilvopastoriles In order to generate an Edaphogeomorphological model to be used for the identification of management requirements of soils, a study was carried out in the area of the Cañada La Gorda watershed, Machiques Colon, Zulia State with a tropical climate, subsumid conditions with a growing period of 230 days, an Ustic soil moisture and Isohypertermic regimes. The soils factorial equation approach was used for the analisis and description of the factors of soil formation throughout a soil catena. Relief was characterized through aerial photographs, satellite images and systematic checks of transects drawn in the sense of surface runoff and also taking into account geomorphological features. Vegetation cover and land use were described and vegetation components were indified by its local names to defined vegetations indicators (VI) for the local conditions. Soils were described and classified according to soil taxonomy and valued by means of a parametric model proposed by Riquier et al, (1970) for determining the productivity index (PI). Two geomorphological landscapes were defined: Hilly and Valley with six positions within the landscapes: hilltop (round or elongated), preserved tableland summit, slopes of high, medium and low tableland and valleys between hills. Representative soils of each position were studied showing a highly advanced degree of edaphogenesis with PI values below 8% in all positions except the valleys with a PI of 13%. Land use type is based on semi intensive pasturing of introduced forage species, with a vegetation of brushwood and scattered shrubs, with some trees relicts of woods affected by timber extraction and turn to grassland Eight VI were identified, highly associated to local physical and hidrological soil conditions. The enormous impact of human activity on soils and vegetation as shown by active erosion processes and absence of wooded areas and the low productivity of livestock systems reported for the area, indicates the necessity of a reorientation of the present land use introducing alternatives like the incorporation of protective woods and agrosilvopastoral management systems.

Ecoturismo Subacuático. Aplicación al Caso Particular de la Evaluación de un Sendero Submarino en las Costas de Maitencillo, V Región, Valparaíso, Chile.

La siguiente tesis nace a partir de la implementación de la actividad de buceo deportivo en el año 2002 en la Caleta de pescadores artesanales de Maitencillo, quinta región, Chile. Los pescadores administran, para su explotación, un tipo de área marina protegida llamada área de manejo y explotación de recursos bentónicos (AMERB), por lo que se hizo indispensable implementar un Plan de Gestión para el buceo deportivo con la finalidad de regular y controlar la actividad para proteger los recursos existentes en el área, sus ecosistemas y su biodiversidad. Para ello, se construyó un Sendero Submarino con 3 circuitos para el buceo. La tesis contextualiza el desarrollo y ejecución del proyecto y evalúa el Plan de Gestión y la utilización del Sendero Submarino y, para ello, se desarrolla una metodología para el cálculo de la capacidad de carga del Sendero y para la evaluación del Plan de Gestión, a través de indicadores biofísicos, socioeconómicos y de gobernabilidad. Esta tesis plantea un modelo de desarrollo a partir del principio del Ecodesarrollo, destacando, principalmente, que es la comunidad local de pescadores artesanales quien ejecuta y administra el proyecto, sin necesidad de recurrir a operadores privados de buceo y, por tanto, logrando para si los beneficios y el control de la actividad; por otro lado, destaca también que es una actividad a baja escala, con bajos impactos ambientales y por tanto sustentable y replicable para otras organizaciones de pescadores. Los resultados indican que con la información biofísica obtenida se pueden tomar medidas de gestión para la actividad de buceo de acuerdo al estado de los recursos bentónicos principales explotados por los pescadores, ya sea restringiendo la actividad, o suspendiéndola por algún período. Los resultados muestran que la cantidad de buceadores que utilizan el Sendero Submarino está muy por debajo de la capacidad de carga calculada, lo que hace que la presión de esta actividad sobre los recursos y el ecosistema marino sea muy baja. Los resultados socioeconómicos y de gobernabilidad indican que esta actividad ha generado ingresos adicionales para los pescadores que trabajan en esta actividad y también ha generado nuevos puestos de trabajo; además, la organización sindical se ha ido fortaleciendo al crear la comisión de ecoturismo y dar responsabilidades a otros pescadores en la gestión de la actividad. La tesis confirma la hipótesis de que “Los Senderos Submarinos pueden ser una forma de turismo sostenible en áreas marinas protegidas”. The present thesis is based on the implementation of scuba diving activities in the year 2002, in the local fishing cove of Maitencillo, Fifth region, Chile. The local fishermen manage the exploitation process of a type of protected marine area called benthic resource management and exploitation area (BRMEA); for this reason, the implementation of a Management programme for scuba diving to regulate and control this activity to protect the resources in the area, their ecosystems and biodiversity, became a priority. In 2002, an Underwater Path was built, with three tracks for scuba diving practice. This thesis contextualises the development and implementation of the Management programme and the use of the Underwater Path. It also develops a methodology to calculate the payload for the Underwater Path, and to assess the Management project through biophysical, economic and governance indicators. This thesis also proposes a development model by the principle of Sustainable development, with the local community of fishermen implementing and handling the project, without the intervention of private diving agencies, generating economic and social benefits, keeping environmental impact to a minimum level, and making it sustainable and worth emulating for other fishermen organisations. The results indicate, through the biophysical data obtained, that it is possible to take managerial measures regarding the scuba diving activity, according to the state of the most exploited benthic resources, by either restricting the activity or plainly suspending it for some time. The results show that the amount of divers using the Underwater path are way below the calculated payload, which keeps the strain that scuba diving causes on these resources and marine ecosystem to the minimum. The results of the socioeconomic and governance issues indicate that this activity has generated additional profits for the fishermen working in this activity, and also new jobs. Furthermore, the syndicate has strengthened thanks to the creation of an ecotourism commission and by assigning other local fishermen with responsibilities in the organisation of the activity. The thesis confirms the hypothesis that states, “Underwater Paths can become a form of sustainable tourism in protected marine areas”