854 resultados para HYDROBENZOFURAN SKELETON


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P2-2 NAC-MYB-BASED TRANSCRIPCIONAL NETWORK INVOLVED IN THE REGULATION OF PHENYLALANINE BIOSYNTHESIS IN P. PINASTER Mª Belén Pascual, Rafael A. Cañas, Blanca Craven-Bartle, Francisco M. Cánovas and Concepción Ávila Departamento de Biología Molecular y Bioquímica. Facultad de Ciencias. Universidad de Málaga. Campus de teatinos s/n, Málaga, Spain Email: cavila@uma.es Conifer trees divert large quantities of carbon into the biosynthesis of phenylpropanoids, particularly to generate lignin, an important constituent of wood. Since phenylalanine is the precursor for phenylpropanoid biosynthesis, the precise regulation of phenylalanine synthesis and use should occur simultaneously. This crucial pathway is finely regulated primarily at the transcriptional level. Transcriptome analyses indicate that the transcription factors (TFs) preferentially expressed during wood formation in plants belong to the MYB and NAC families. Craven-Bartle et al. (2013) have shown that Myb8 is a candidate regulator of key genes in phenylalanine biosynthesis involved in the supply of the phenylpropane carbon skeleton necessary for lignin biosynthesis. This TF is able to bind AC elements present in the promoter regions of these genes to activate transcription. In Arabidopsis, the transcriptional network controlling secondary cell wall involves NAC-domain regulators operating upstream Myb transcription factors. We have identified in the P. pinaster genome three NAC proteins as potential candidates to be involved in vascular development. One of them, PpNAC1 is expressed both in xylem and compression wood from adult trees and has been thoroughly characterized. Its role upstream the transcriptional network involving Myb8 will be discussed. The understanding of the transcriptional regulatory network associated to phenylpropanoids and lignin biosynthesis in conifers is crucial for future applications in tree improvement and sustainable forest management.

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This paper investigates the robust and accurate capture of human joint poses and bio-kinematic movements for exercise monitoring in real-time tele-rehabilitation applications. Recently developed model-based estimation ideas are used to improve the accuracy, robustness, and real-time characteristics considered vital for applications, where affordability and domestic use are the primary focus. We use the spatial diversity of the arbitrarily positioned Microsoft Kinect receivers to improve the reliability and promote the uptake of the concept. The skeleton-based information is fused to enhance accuracy and robustness, critical for biomedical applications. A specific version of a robust Kalman filter (KF) in a linear framework is employed to ensure superior estimator convergence and real-time use, compared to other commonly used filters. The algorithmic development was conducted in a generic form and computer simulations were conducted to verify our assertions. Hardware implementations were carried out to test the viability of the proposed state estimator in terms of the core requirements of reliability, accuracy, and real-time use. Performance of the overall system implemented in an information fusion context was evaluated against the commercially available and industry standard Vicon system for different exercise routines, producing comparable results with much less infrastructure and financial investment.

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The use of instability devices and exercises to train the core musculature is an essential feature of many training centres and programs. It was the intent of this position stand to provide recommendations regarding the role of instability in resistance training programs designed to train the core musculature. The core is defined as the axial skeleton and all soft tissues with a proximal attachment originating on the axial skeleton, regardless of whether the soft tissue terminates on the axial or appendicular skeleton. Core stability can be achieved with a combination of muscle activation and intra-abdominal pressure. Abdominal bracing has been shown to be more effective than abdominal hollowing in optimizing spinal stability. When similar exercises are performed, core and limb muscle activation are reported to be higher under unstable conditions than under stable conditions. However, core muscle activation that is similar to or higher than that achieved in unstable conditions can also be achieved with ground-based free-weight exercises, such as Olympic lifts, squats, and dead lifts. Since the addition of unstable bases to resistance exercises can decrease force, power, velocity, and range of motion, they are not recommended as the primary training mode for athletic conditioning. However, the high muscle activation with the use of lower loads associated with instability resistance training suggests they can play an important role within a periodized training schedule, in rehabilitation programs, and for nonathletic individuals who prefer not to use ground-based free weights to achieve musculoskeletal health benefits.

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Training of the trunk or core muscles for enhanced health, rehabilitation, and athletic performance has received renewed emphasis. Instability resistance exercises have become a popular means of training the core and improving balance. Whether instability resistance training is as, more, or less effective than traditional ground-based resistance training is not fully resolved. The purpose of this review is to address the effectiveness of instability resistance training for athletic, nonathletic, and rehabilitation conditioning. The anatomical core is defined as the axial skeleton and all soft tissues with a proximal attachment on the axial skeleton. Spinal stability is an interaction of passive and active muscle and neural subsystems. Training programs must prepare athletes for a wide variety of postures and external forces, and should include exercises with a destabilizing component. While unstable devices have been shown to be effective in decreasing the incidence of low back pain and increasing the sensory efficiency of soft tissues, they are not recommended as the primary exercises for hypertrophy, absolute strength, or power, especially in trained athletes. For athletes, ground-based free-weight exercises with moderate levels of instability should form the foundation of exercises to train the core musculature. Instability resistance exercises can play an important role in periodization and rehabilitation, and as alternative exercises for the recreationally active individual with less interest or access to ground-based free-weight exercises. Based on the relatively high proportion of type I fibers, the core musculature might respond well to multiple sets with high repetitions (e.g., >15 per set); however, a particular sport may necessitate fewer repetitions.

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Sachant que plusieurs maladies entrainent des lésions qui ne sont pas toujours observables à l’oeil, cette étude préliminaire en paléopathologie humaine utilise une approche complémentaire issue de l’imagerie médicale, le ct-scan, afin de fournir des diagnostics plus précis. L’objectif est donc de tester ici l’efficacité et les limites de l’analyse scanographique durant l’analyse de spécimens archéologiques. Un échantillon de 55 individus a été sélectionné à partir de la collection ostéologique provenant du cimetière protestant St. Matthew (ville de Québec, 1771 – 1860). Une analyse macroscopique et scanographique complète a alors été effectuée sur chaque squelette. Les observations macroscopiques ont consisté à enregistrer une dizaine de critères standardisés par la littérature de référence en lien avec des manifestations anormales à la surface du squelette. Les ct-scans ont été réalisés à l’Institut National de la Recherche Scientifique de la Ville de Québec avec un tomodensitomètre Somatom de Siemens (définition AS+ 128). Les données scanographiques ont permis d’enregistrer une série de critères complémentaires sur la structure interne de l’os (amincissement/épaississement de la corticale, variation de densité, etc.) Selon la méthode du diagnostic différentiel, des hypothèses ou diagnostics ont été proposés. Ils sont principalement basés sur les critères diagnostiques mentionnés dans les manuels de référence en paléopathologie, mais aussi à l’aide de la littérature clinique et l’expertise de médecins. Les résultats présentés ici supportent que: 1) Dans 43% des cas, les données scanographiques ont apporté des informations essentielles dans la diagnose pathologique. Cette tendance se confirme en fonction de certaines maladies, mais pas d’autres, car certains diagnostics ne peuvent se faire sans la présence de tissus mous. 2) La distribution spatiale de la plupart des lésions varie selon les régions anatomiques, aussi bien en macroscopie qu’en scanographie. 3) Certains types de maladie semblent associés à l’âge et au sexe, ce qui est conforté par la littérature. 4) Cette recherche démontre aussi que le processus de diagnose nécessite, dans 38% des cas, une analyse complémentaire (ex. histologie, scintigraphie, radiographie) pour préciser le diagnostic final.

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Kainic acid has been used for nearly 50 years as a tool in neuroscience due to its pronounced neuroexcitatory properties. However, the significant price increase of kainic acid resulting from the disruption in the supply from its natural source, the alga Digenea Simplex, as well as inefficient synthesis of kainic acid, call for the exploration of functional mimics of kainic acid that can be synthesized in a simpler way. Aza kainoids analog could be one of them. The unsubstituted aza analog of kainoids has demonstrates its ability as an ionotropic glutamate receptor agonist and showed affinity in the chloride dependent glutamate (GluCl) binding site. This opened a question of the importance of the presence of one nitrogen or both nitrogens in the aza kainoid analogs for binding to glutamate receptors. Therefore, two different pyrrolidine analogs of kainic acid, trans-4-(carboxymethyl)pyrrolidine-3-carboxylic acid and trans-2-carboxy-3-pyrrolidineacetic acid, were synthesized through multi-step sequences. The lack of the affinity of both pyrrolidine analogs in GluCl binding site indicated that both nitrogens in aza kainoid analogs are involved in hydrogen bonding with receptors, significantly enhancing their affinity in GluCl binding site. Another potential functional mimic of kainic acid is isoxazolidine analogs of kainoids whose skeleton can be constituted directly via a 1, 3 dipolar cycloaddition as the key step. The difficulty in synthesizing N-unsubstituted isoxazolidines when applying such common protecting groups as alkyl, phenyl and benzyl groups, and the requirement of a desired enantioselectivity due to the three chiral ceneters in kainic acid, pose great challenges. Hence, several different protected nitrones were studied to establish that diphenylmethine nitrone may be a good candidate as the dipole in that the generated isoxazolidines can be deprotected in mild conditions with high yields. Our investigations also indicated that the exo/endo selectivity of the 1, 3 dipolar cycloaddition can be controlled by Lewis acids, and that the application of a directing group in dipolarophiles can accomplish a satisfied enantioselectivity. Those results demonstrated the synthesis of isoxazoldines analogs of kainic acid is very promising.

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In order to reduce serious health incidents, individuals with high risks need to be identified as early as possible so that effective intervention and preventive care can be provided. This requires regular and efficient assessments of risk within communities that are the first point of contacts for individuals. Clinical Decision Support Systems CDSSs have been developed to help with the task of risk assessment, however such systems and their underpinning classification models are tailored towards those with clinical expertise. Communities where regular risk assessments are required lack such expertise. This paper presents the continuation of GRiST research team efforts to disseminate clinical expertise to communities. Based on our earlier published findings, this paper introduces the framework and skeleton for a data collection and risk classification model that evaluates data redundancy in real-time, detects the risk-informative data and guides the risk assessors towards collecting those data. By doing so, it enables non-experts within the communities to conduct reliable Mental Health risk triage.

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A rapid and efficient method to identify the weak points of the complex chemical structure of low band gap (LBG) polymers, designed for efficient solar cells, when submitted to light exposure is reported. This tool combines Electron Paramagnetic Resonance (EPR) using the 'spin trapping method' coupled with density functional theory modelling (DFT). First, the nature of the short life-time radicals formed during the early-stages of photo-degradation processes are determined by a spin-trapping technique. Two kinds of short life-time radical (R and R′O) are formed after 'short-duration' illumination in an inert atmosphere and in ambient air, respectively. Second, simulation allows the identification of the chemical structures of these radicals revealing the most probable photochemical process, namely homolytical scission between the Si atom of the conjugated skeleton and its pendent side-chains. Finally, DFT calculations confirm the homolytical cleavage observed by EPR, as well as the presence of a group that is highly susceptible to photooxidative attack. Therefore, the synergetic coupling of a spin trapping method with DFT calculations is shown to be a rapid and efficient method for providing unprecedented information on photochemical mechanisms. This approach will allow the design of LBG polymers without the need to trial the material within actual solar cell devices, an often long and costly screening procedure.

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Sachant que plusieurs maladies entrainent des lésions qui ne sont pas toujours observables à l’oeil, cette étude préliminaire en paléopathologie humaine utilise une approche complémentaire issue de l’imagerie médicale, le ct-scan, afin de fournir des diagnostics plus précis. L’objectif est donc de tester ici l’efficacité et les limites de l’analyse scanographique durant l’analyse de spécimens archéologiques. Un échantillon de 55 individus a été sélectionné à partir de la collection ostéologique provenant du cimetière protestant St. Matthew (ville de Québec, 1771 – 1860). Une analyse macroscopique et scanographique complète a alors été effectuée sur chaque squelette. Les observations macroscopiques ont consisté à enregistrer une dizaine de critères standardisés par la littérature de référence en lien avec des manifestations anormales à la surface du squelette. Les ct-scans ont été réalisés à l’Institut National de la Recherche Scientifique de la Ville de Québec avec un tomodensitomètre Somatom de Siemens (définition AS+ 128). Les données scanographiques ont permis d’enregistrer une série de critères complémentaires sur la structure interne de l’os (amincissement/épaississement de la corticale, variation de densité, etc.) Selon la méthode du diagnostic différentiel, des hypothèses ou diagnostics ont été proposés. Ils sont principalement basés sur les critères diagnostiques mentionnés dans les manuels de référence en paléopathologie, mais aussi à l’aide de la littérature clinique et l’expertise de médecins. Les résultats présentés ici supportent que: 1) Dans 43% des cas, les données scanographiques ont apporté des informations essentielles dans la diagnose pathologique. Cette tendance se confirme en fonction de certaines maladies, mais pas d’autres, car certains diagnostics ne peuvent se faire sans la présence de tissus mous. 2) La distribution spatiale de la plupart des lésions varie selon les régions anatomiques, aussi bien en macroscopie qu’en scanographie. 3) Certains types de maladie semblent associés à l’âge et au sexe, ce qui est conforté par la littérature. 4) Cette recherche démontre aussi que le processus de diagnose nécessite, dans 38% des cas, une analyse complémentaire (ex. histologie, scintigraphie, radiographie) pour préciser le diagnostic final.

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Detta projekt beställdes av lekredskapstillverkaren HAGS i Aneby. De behövde en docka i en 4-årings storlek till sin produktutveckling. Det företaget frågar efter är en så antropomorfiskt korrekt docka som möjligt med framför allt korrekta proportioner på huvud och lemmar. Denna docka skulle användas för att fastställa måtten på nya produkter och för säkrare säkerhetstester. För att dockan skulle vara könsneutral så fick både dockan och projektet namnet KIM. Projektet har gått igenom flera faser. Först specificerades vad som skulle göras och vilka delar som var viktigast att fokusera på. Sedan löstes hur dockans alla leder skall fungera, hur ytan skall behandlas och hur allting skall monteras. Den huvudsakliga metoden för framtagandet av koncept och slutresultat har varit en iterativ process där lösningar har tagits fram, testats, ändrats och testats igen. Det som har framställts i projektet är koncept för ett helt skelett som med lättare modifieringar kan användas till flera applikationer. Delarna i skelettet är konstruerade efter en ”One Size Fits All” princip där ett litet antal delar kan användas till flera funktionsområden. En del arbete återstår, huvudet har inte fått sin slutgiltiga konstruktion och både händer och fötter behöver gå igenom ett par designfaser för att färdigställas. Det som framför allt återstår av projektet är tillverkning av alla delar, vilket innebär mycket arbete med en 3d-skrivare för alla leder och skulptering för hand för alla mjukdelar.

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Oleuropein, the main phenolic compound of olive leaves, exhibits a unique blend of biological activities and has been shown to locate itself at the oil-water (O/W) interface. This behavior could influence the physico-chemical properties of dispersed systems such as emulsions. In this work, we study the effect of the microenvironment (vacuum, water, and triolein-water) on the conformational preferences of oleuropein using molecular dynamics (MD) simulations at 300K for at least 30ns. The seven torsions that describe the flexible skeleton of oleuropein were monitored together with the distance between the glucose (Glu) and hydroxytyrosol (Hyd) moieties (dglu-hyd) of the molecule. The obtained trajectories demonstrated that oleuropein adopts different conformations that depend on the environment. The preferential conformers in each system were analyzed for their molecular geometry and internal energy. In vacuum, the oleuropein preferential conformation is tight with the glucose moiety in close proximity with the hydroxytyrosol moiety. In water, oleuropein preferential conformers presented large differences in their structural properties, varying from a close like U form, and a semi-opened form, to an opened form characterized by high fluctuations in dglu-hyd values. In a triolein-water system, oleuropein tends to adopt a more open form where the glucose moiety could be approximately aligned with the hydroxytyrosol and elenolic acid moieties. Based on a calculation at the HF/6-31G* level, these flexibilities of oleuropein required energy of 19.14kcal/mol in order to adopt the conformation between water and triolein-water system. A radial distribution function (RDF) analysis showed that specific hydroxyl groups of Hyd and Glu interact with water molecules, enabling us to understand the amphiphilic character of oleuropein at the triolein-water interface. MD calculations together with interfacial tension measurements revealed that the oleuropein binding at O/W interface is an enthalpy driven mechanism.

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Abstract : Although concrete is a relatively green material, the astronomical volume of concrete produced worldwide annually places the concrete construction sector among the noticeable contributors to the global warming. The most polluting constituent of concrete is cement due to its production process which releases, on average, 0.83 kg CO[subscript 2] per kg of cement. Self-consolidating concrete (SCC), a type of concrete that can fill in the formwork without external vibration, is a technology that can offer a solution to the sustainability issues of concrete industry. However, all of the workability requirements of SCC originate from a higher powder content (compared to conventional concrete) which can increase both the cost of construction and the environmental impact of SCC for some applications. Ecological SCC, Eco-SCC, is a recent development combing the advantages of SCC and a significantly lower powder content. The maximum powder content of this concrete, intended for building and commercial construction, is limited to 315 kg/m[superscript 3]. Nevertheless, designing Eco-SCC can be challenging since a delicate balance between different ingredients of this concrete is required to secure a satisfactory mixture. In this Ph.D. program, the principal objective is to develop a systematic design method to produce Eco-SCC. Since the particle lattice effect (PLE) is a key parameter to design stable Eco-SCC mixtures and is not well understood, in the first phase of this research, this phenomenon is studied. The focus in this phase is on the effect of particle-size distribution (PSD) on the PLE and stability of model mixtures as well as SCC. In the second phase, the design protocol is developed, and the properties of obtained Eco-SCC mixtures in both fresh and hardened states are evaluated. Since the assessment of robustness is crucial for successful production of concrete on large-scale, in the final phase of this work, the robustness of one the best-performing mixtures of Phase II is examined. It was found that increasing the volume fraction of a stable size-class results in an increase in the stability of that class, which in turn contributes to a higher PLE of the granular skeleton and better stability of the system. It was shown that a continuous PSD in which the volume fraction of each size class is larger than the consecutive coarser class can increase the PLE. Using such PSD was shown to allow for a substantial increase in the fluidity of SCC mixture without compromising the segregation resistance. An index to predict the segregation potential of a suspension of particles in a yield stress fluid was proposed. In the second phase of the dissertation, a five-step design method for Eco-SCC was established. The design protocol started with the determination of powder and water contents followed by the optimization of sand and coarse aggregate volume fractions according to an ideal PSD model (Funk and Dinger). The powder composition was optimized in the third step to minimize the water demand while securing adequate performance in the hardened state. The superplasticizer (SP) content of the mixtures was determined in next step. The last step dealt with the assessment of the global warming potential of the formulated Eco-SCC mixtures. The optimized Eco-SCC mixtures met all the requirements of self-consolidation in the fresh state. The 28-day compressive strength of such mixtures complied with the target range of 25 to 35 MPa. In addition, the mixtures showed sufficient performance in terms of drying shrinkage, electrical resistivity, and frost durability for the intended applications. The eco-performance of the developed mixtures was satisfactory as well. It was demonstrated in the last phase that the robustness of Eco-SCC is generally good with regards to water content variations and coarse aggregate characteristics alterations. Special attention must be paid to the dosage of SP during batching.

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Hydrogen is considered as an appealing alternative to fossil fuels in the pursuit of sustainable, secure and prosperous growth in the UK and abroad. However there exists a persisting bottleneck in the effective storage of hydrogen for mobile applications in order to facilitate a wide implementation of hydrogen fuel cells in the fossil fuel dependent transportation industry. To address this issue, new means of solid state chemical hydrogen storage are proposed in this thesis. This involves the coupling of LiH with three different organic amines: melamine, urea and dicyandiamide. In principle, thermodynamically favourable hydrogen release from these systems proceeds via the deprotonation of the protic N-H moieties by the hydridic metal hydride. Simultaneously hydrogen kinetics is expected to be enhanced over heavier hydrides by incorporating lithium ions in the proposed binary hydrogen storage systems. Whilst the concept has been successfully demonstrated by the results obtained in this work, it was observed that optimising the ball milling conditions is central in promoting hydrogen desorption in the proposed systems. The theoretical amount of 6.97 wt% by dry mass of hydrogen was released when heating a ball milled mixture of LiH and melamine (6:1 stoichiometry) to 320 °C. It was observed that ball milling introduces a disruption in the intermolecular hydrogen bonding network that exists in pristine melamine. This effect extends to a molecular level electron redistribution observed as a function of shifting IR bands. It was postulated that stable phases form during the first stages of dehydrogenation which contain the triazine skeleton. Dehydrogenation of this system yields a solid product Li2NCN, which has been rehydrogenated back to melamine via hydrolysis under weak acidic conditions. On the other hand, the LiH and urea system (4:1 stoichiometry) desorbed approximately 5.8 wt% of hydrogen, from the theoretical capacity of 8.78 wt% (dry mass), by 270 °C accompanied by undesirable ammonia and trace amount of water release. The thermal dehydrogenation proceeds via the formation of Li(HN(CO)NH2) at 104.5 °C; which then decomposes to LiOCN and unidentified phases containing C-N moieties by 230 °C. The final products are Li2NCN and Li2O (270 °C) with LiCN and Li2CO3 also detected under certain conditions. It was observed that ball milling can effectively supress ammonia formation. Furthermore results obtained from energetic ball milling experiments have indicated that the barrier to full dehydrogenation between LiH and urea is principally kinetic. Finally the dehydrogenation reaction between LiH and dicyandiamide system (4:1 stoichiometry) occurs through two distinct pathways dependent on the ball milling conditions. When ball milled at 450 RPM for 1 h, dehydrogenation proceeds alongside dicyandiamide condensation by 400 °C whilst at a slower milling speed of 400 RPM for 6h, decomposition occurs via a rapid gas desorption (H2 and NH3) at 85 °C accompanied by sample foaming. The reactant dicyandiamide can be generated by hydrolysis using the product Li2NCN.

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O presente trabalho consiste na apresentação da diversidade de seláceos do Farol das Lagostas pertencente à Bacia do Cuanza. Os condrichthyes são componentes comuns da fauna aquática desde o Paleozoico. No entanto, devido à natureza cartilaginosa do esqueleto, o registo paleontológico dos mesmos restringe-se, basicamente, às partes mineralizadas como dentes, dentículos dérmicos e espinhas cefálicas O lugar Farol das Lagostas contém uma fauna de seláceos relativamente rica e variada, representada neste estudo por cerca de 1.000 dentes isolados, além de outras peças esqueléticas. Foram identificados e descritos quarenta e quatro taxa, pertencentes às seguintes ordens: Hexanchiformes, Squaliformes, Pristiophoriformes, Squatiniformes, Lamniformes, Carcharhiniformes, Rajiformes e Myliobatiformes. Trata-se de uma diversidade faunística cuja maioria dos géneros possui representantes atuais. Os Carcharhiniformes representam 53% dos taxa identificados. Estão representadas as famílias Scyliorhinidae, Triakidae, Hemigaleidae, Carcharhinidae e Sphyrnidae, com 18 espécies. Verifica-se a presença de formas bentónicas e nectónicas que indicam condições térmicas moderadas, quentes e tropicais a temperadas, que habitam frequentemente zonas costeiras. A relação faunística identificada corresponde a uma zona litoral. Ora, considerando o conjunto de dados em face da presença de fauna pelágica e dos grandes predadores, especialmente Isurus hastalis e Carcharocles megalodon, podemos admitir que existisse então um Golfo relativamente largo de uma faixa Atlântica aberta (Antunes & Balbino, 2004); ABSTRACT: Selachians from Farol the Lagostas (Cuanza Basin, Angola) The present work consists of the presentation of diversity of selachians from Farol das Lagostas, which belongs to the Cuanza Basin. The condrichthyes are a part of aquatic fauna since the Paleozoic. However, due to the nature of the cartilaginous skeleton, the paleontological registration of the same ones is restricted to mineralized parts like teeth, dermal denticles and cephalic spines. The of formation, Farol das Lagostas has a fauna of selachian relatively rich and diverse, represented in this study for around 1.000 isolated teeth, and other skeletal parts. Forty four taxa were described and identified, belonging to the following orders: Hexanchiformes, Squaliformes, Pristiophoriformes, Squatiniformes, Lamniformes, Carcharhiniformes, Rajiformes e Myliobatiformes. It’s a faunal diversity whose most genera have current representatives. The Carcharhiniformes represente 53% of the identified taxa. The represented families are Scyliorhinidae, Triakidae, Hemigaleidae, Carcharhinidae e Sphyrnidae, with 18 species. There is presence of benthic and nektonic forms that indicate moderate thermal conditions, the temperate and tropical hot, often inhabit coastal areas. The identified faunal relation corresponds to a coastal zone. So, considering the data set, due to the presence of pelagic fauna and large predators, especially Isurus hastalis e Carcharocles megalodon, we can admit that then there was a relatively large Gulf of open Atlantic range (Antunes & Balbino, 2004).