Identification of cadmium-induced mutations in a mammalian cell line

Epidemiological studies have shown cadmium to induce cancer in humans, while experimental studies have proven this metal to be a potent tumor inducer in animals. However, cadmium appears nonmutagenic in most prokaryotic and eukaryotic mutagenesis assays. In this study, we present the identification of mutations in normal rat kidney cells infected with the mutant MuSVts110 retrovirus (6m2 cells) as a result of treatment with cadmium chloride. The detection of these mutations was facilitated by the use of a novel mutagenesis assay established in this laboratory. The 6m2 reversion assay is a positive selection system based on the conditional expression of the MuSVts110 v-mos gene. In MuSVts110 the gag and mos genes are fused out of frame, thus the translation of the v-mos sequence requires a frameshift in the genomic RNA. In 6m2 cells this frameshift is accomplished by the temperature-dependent splicing of the primary MuSVts110 transcript. Splicing of MuSVts110, which is mediated by cis-acting sequences, occurs when 6m2 cells are grown at 33$\sp\circ$C and below, but not at 39$\sp\circ$C. Therefore, 6m2 cells appear transformed at low growth temperatures, but take on a morphologically normal appearance when grown at high temperatures. The treatment of 6m2 cells with cadmium chloride resulted in the outgrowth of a number of cells that reverted to the transformed state at high growth temperatures. Analysis of the viral proteins expressed in these cadmium-induced 6m2 revertants suggested that they contained mutations in their MuSVts110 DNA. Sequencing of the viral DNA from three revertants that constitutively expressed the P85$\sp{gag{-}mos}$ transforming protein revealed five different mutations. The Cd-B2 revertant contained three of those mutations: an A-to-G transition 48 bases downstream of the MuSVts110 3$\sp\prime$ splice site, plus a G-to-T and an A-to-T transversion 84 and 100 bases downstream of the 5$\sp\prime$ splice site, respectively. The Cd-15-5 revertant also contained a point mutation, a T-to-C transition 46 bases downstream of the 5$\sp\prime$ splice site, while Cd-10-5 contained a three base deletion of MuSVts110 11 bases upstream of the 3$\sp\prime$ splice site. A fourth revertant, Cd-10, expressed a P100$\sp{gag{-}mos}$ transforming protein, and was found to have a two base deletion. This deletion accomplished the frameshift necessary for v-mos expression, but did not alter MuSVts110 RNA splicing and the expression of p85$\sp{gag{-}mos}.$ Lastly, sequencing of the MuSVts110 DNA from three spontaneous revertants revealed the same G to T transversion in each one. This was the same mutation that was found in the Cd-B2 revertant. These findings provide the first example of mutations resulting from exposure to cadmium and suggest, by the difference in each mutation, the complexity of the mechanism utilized by cadmium to induce DNA damage. ^

The Glucose-Dependent Insulinotropic Polypeptide Receptor: A Novel Target for Neuroendocrine Tumor Imaging-First Preclinical Studies

A new family of peptide receptors, the incretin receptor family, overexpressed on many neuroendocrine tumors (NETs) is of great importance because it may enable the in vivo peptide-based receptor targeting of a category of NETs that does not express the somatostatin receptor. Impressive in vivo diagnostic data were published for glucagonlike peptide 1 receptor-targeting radiopeptides. Recently, promising in vitro data have appeared for the second member of the incretin family, the glucose-dependent insulinotropic polypeptide (GIP) receptor. This prompted us to develop and evaluate a new class of radioligands with the potential to be used for the in vivo targeting of GIP receptor-positive tumors. METHODS GIP(1-42) was modified C-terminally, and the truncated peptides [Lys(30)(aminohexanoic acid [Ahx]-DOTA)]GIP(1-30)NH2 (EG1), [Lys(16)(Ahx-DOTA)]GIP(1-30)NH2 (EG2), and [Nle(14), Lys(30)(Ahx-DOTA)]GIP(1-30)NH2 (EG4) were conjugated with Ahx-DOTA via the Lys(16) and Lys(30) side chains. Their inhibitory concentration of 50% (IC50) was determined using [(125)I-Tyr(10)]GIP(1-30) as radioligand and GIP(1-30) as control peptide. The DOTA conjugates were labeled with (111)In and (68)Ga. In vitro evaluation included saturation and internalization studies using the pancreatic endocrine cell line INR1G9 transfected with the human GIP receptor (INR1G9-hGIPr). The in vivo evaluation consisted of biodistribution and PET imaging studies on nude mice bearing INR1G9-hGIPr tumors. RESULTS Binding studies (IC50 and saturation studies) showed high affinity toward GIP receptor for the GIP conjugates. Specific in vitro internalization was found, and almost the entire cell-associated activity was internalized (>90% of the cell-bound activity), supporting the agonist potency of the (111)In-vectors. (111)In-EG4 and (68)Ga-EG4 were shown to specifically target INR1G9-hGIPr xenografts, with tumor uptake of 10.4% ± 2.2% and 17.0% ± 4.4% injected activity/g, 1 h after injection, respectively. Kidneys showed the highest uptake, which could be reduced by approximately 40%-50% with a modified-fluid-gelatin plasma substitute or an inhibitor of the serine protease dipeptidyl peptidase 4. The PET images clearly visualized the tumor. CONCLUSION The evaluation of EG4 as a proof-of-principle radioligand indicated the feasibility of imaging GIP receptor-positive tumors. These results prompt us to continue the development of this family of radioligands for imaging of a broad spectrum of NETs.

The first major incision of the Swiss Deckenschotter landscape

The Swiss Deckenschotter (“cover gravels”) is the oldest Quaternary units in the northern Swiss Alpine Foreland. They are a succession of glaciofluvial gravel layers intercalated with glacial and/or overbank deposits. This lithostratigraphic sequence is called Deckenschotter because it “covers” Molasse or Mesozoic bedrock and forms mesa-type hill-tops. Deckenschotter occurs both within and beyond the extent of the Last Glacial Maximum glaciers. The Swiss Deckenschotter consist of two sub-units: Höhere (Higher) and Tiefere (Lower) Deckenschotter. Although the Höhere Deckenschotter sub-unit (HDS) is topographically higher than the Tiefere Deckenschotter, it is older. The only available age for the Swiss Deckenschotter is 2.5–1.8 Ma based on mammal remains found in HDS at the Irchel site. In this study, we present an exposure age for the topographically lowest HDS, calculated from a cosmogenic 10Be depth-profile. Our results show that the first phase of the Deckenschotter glaciations in the Swiss Alps terminated at least 1,020+80−120 ka ago, which is indicated by a significant fluvial incision. This line of evidence seems to be close to synchronous with the beginning of the Mid-Pleistocene Revolution, when the frequency of the glacial-interglacial cyclicity changed from 41 to 100 ka and the amplitude from low to high, between marine isotope stages 23 and 22.

Genetic stability of Schmallenberg virus in vivo during an epidemic, and in vitro, when passaged in the highly susceptible porcine SK-6 cell line

Schmallenberg virus (SBV), an arthropod-borne orthobunyavirus was first detected in 2011 in cattle suffering from diarrhea and fever. The most severe impact of an SBV infection is the induction of malformations in newborns and abortions. Between 2011 and 2013 SBV spread throughout Europe in an unprecedented epidemic wave. SBV contains a tripartite genome consisting of the three negative-sense RNA segments L, M, and S. The virus is usually isolated from clinical samples by inoculation of KC (insect) or BHK-21 (mammalian) cells. Several virus passages are required to allow adaptation of SBV to cells in vitro. In the present study, the porcine SK-6 cell line was used for isolation and passaging of SBV. SK-6 cells proved to be more sensitive to SBV infection and allowed to produce higher titers more rapidly as in BHK-21 cells after just one passage. No adaptation was required. In order to determine the in vivo genetic stability of SBV during an epidemic spread of the virus the nucleotide sequence of the genome from seven SBV field isolates collected in summer 2012 in Switzerland was determined and compared to other SBV sequences available in GenBank. A total of 101 mutations, mostly transitions randomly dispersed along the L and M segment were found when the Swiss isolates were compared to the first SBV isolated late 2011 in Germany. However, when these mutations were studied in detail, a previously described hypervariable region in the M segment was identified. The S segment was completely conserved among all sequenced SBV isolates. To assess the in vitro genetic stability of SBV, three isolates were passage 10 times in SK-6 cells and sequenced before and after passaging. Between two and five nt exchanges per genome were found. This low in vitro mutation rate further demonstrates the suitability of SK-6 cells for SBV propagation.

Long-term evaluation of Class II subdivision treatment with unilateral maxillary first molar extraction

OBJECTIVE To evaluate the long-term effects of asymmetrical maxillary first molar (M1) extraction in Class II subdivision treatment. MATERIALS AND METHODS Records of 20 Class II subdivision whites (7 boys, 13 girls; mean age, 13.0 years; SD, 1.7 years) consecutively treated with the Begg technique and M1 extraction, and 15 untreated asymmetrical Class II adolescents (4 boys, 11 girls; mean age, 12.2 years; SD, 1.3 years) were examined in this study. Cephalometric analysis and PAR assessment were carried out before treatment (T1), after treatment (T2), and on average 2.5 years posttreatment (T3) for the treatment group, and at similar time points and average follow-up of 1.8 years for the controls. RESULTS The adjusted analysis indicated that the maxillary incisors were 2.3 mm more retracted in relation to A-Pog between T1 and T3 (β  =  2.31; 95% CI; 0.76, 3.87), whereas the mandibular incisors were 1.3 mm more protracted (β  =  1.34; 95% CI; 0.09, 2.59), and 5.9° more proclined to the mandibular plane (β  =  5.92; 95% CI; 1.43, 10.41) compared with controls. The lower lip appeared 1.4 mm more protrusive relative to the subnasale-soft tissue-Pog line throughout the observation period in the treated adolescents (β  =  1.43; 95% CI; 0.18, 2.67). There was a significant PAR score reduction over the entire follow-up period in the molar extraction group (β  =  -6.73; 95% CI; -10.7, -2.7). At T2, 65% of the subjects had maxillary midlines perfectly aligned with the face. CONCLUSIONS Unilateral M1 extraction in asymmetrical Class II cases may lead to favorable occlusal outcomes in the long term without harming the midline esthetics and soft tissue profile.

Evaluation of the LBJ Hospital Ask Your Nurse Advice Line

This retrospective, case-control study investigated the effectiveness of the Houston, Texas Ask Your Nurse Advice Line (AYN) in diverting callers with non-emergent medical conditions away from the emergency department (ED). After asking callers a series of questions, AYN nurses evaluate the medical condition and make recommendations for appropriate care, e.g., home care, see a clinic physician, or visit the ED. To evaluate the AYN, the rate of caller ED visits before accessing the AYN for the first time was compared to the caller ED visit rate afterwards. The pre-post rate change was compared to that of a control group of similar caller age, race, gender, and insurance status drawn from a Harris County Hospital District HCHD database. ^ The treatment group (AYN caller) had a 66% reduction in ED visits after the first AYN call compared to an 18% drop in ED visits among control group subjects during the same time period. Study results were presented to HCHD staff on August 30th, 2007 and recommendations were made for future studies that would provide a basis for policy development. ^

Aberrant regulation of the Src -FAK signaling pathway in the human colon adenocarcinoma cell line HT-29

c-Src, a protein tyrosine kinase (PTK) the specific activity of which is increased $>$20-fold in $\sim$80% of colon tumors and colon tumor cell lines, plays a role in both growth regulation and tumorigenicity of colon tumor cells. To examine the effect of increased c-Src specific activity on colon tumor cells, coumarin-derived tyrosine analog PTK inhibitors were assessed in a standard colon tumor cell line, HT-29. Of the nine compounds tested for inhibiting c-Src activity in a standard immune complex kinase assay from c-Src precipitated from HT-29 cells, the 7,8-dihydroxy-containing compounds daphnetin and fraxetin were most effective, with IC$\sb{50}$s of 0.6 $\pm$ 0.2 mM and 0.6 $\pm$ 0.3 mM, respectively. Treatment of HT-29 cells with daphnetin resulted in inhibition of cell growth in a dose-dependent manner. In contrast, scopoletin, a relatively poor Src inhibitor in vitro, did not inhibit HT-29 cell growth in the concentration range tested. In daphnetin treated cells, a dose-dependent decrease of c-Src activity paralleling cell growth inhibition was also observed; the IC$\sb{50}$ was 0.3 $\pm$ 0.1 mM for c-Src autophosphorylation. In contrast, the IC$\sb{50}$ for c-Src protein level was $>$ 0.6 mM, indicating that the effects of daphnetin were primarily an enzymatic activity of c-Src, rather than protein level in HT-29 cells. These results are the first to demonstrate that c-Src specific activity regulates colon tumor cell growth.^ To elucidate the signaling pathways activated by c-Src in colon tumor cells, the Src family substrate FAK, which has been shown to play a role in both extracellular matrix-dependent cell growth and survival, was examined. Coprecipitation assays showed Src-FAK association in detergent insoluble fractions of both attached and detached HT-29 cells, indicating that Src-FAK association in HT-29 cells is stable and, unlike untransformed cells, not dependent on cell-substratum contact. FAK also coprecipitated with Grb2, an adaptor protein also playing a role in cell proliferation and survival, in both attached and detached HT-29 cells, suggesting that a Src-FAK-Grb2-mediated signaling pathway(s) in HT-29 cells is/are constitutively activated.^ FAK was also analyzed in c-src antisense HT-29 clones AS15 and AS33 in which c-Src is specifically reduced by transfection of an antisense expression vector. FAK protein level is unexpectedly decreased in both AS15 and AS33 cells by 5-fold and 1.5-fold compared to HT-29, respectively, corresponding with the decreased expression of c-Src observed in these cells. FAK protein level was not decreased compared to parental in the c-src "sense" clone S8. Northern blot analyses showed decreased FAK mRNA levels compared to parental in AS15 and AS33, correlating with decreased FAK protein level, indicating that FAK activity in the antisense cells is regulated, at least in part, by altering FAK expression, and that this regulation is Src dependent. Because FAK has been implicated in anoikis, the ability of c-src antisense cells to survive in the absence of cell-substratum contact was examined. Decreased cell survival is seen in both AS15 and AS33, correlating with the decreases in c-Src and FAK levels and tumorigenicity in these cells. These results suggest that at least one mechanism by which activation of c-Src contributes to tumorigenic phenotype of colon tumor cells is by aberrantly promoting a survival signal through unregulated Src-FAK-Grb2 complexes. (Abstract shortened by UMI.) ^

Coordinates of multichannel seismic line GeoB05-004

Major plastered drift sequences were imaged using high-resolution multichannel seismics during R/V Meteor cruises M63/1 and M75/3 south of the Mozambique Channel along the continental margin of Mozambique off the Limpopo River. Detailed seismic-stratigraphic analyses enabled the reconstruction of the onset and development of the modern, discontinuous, eddy-dominated Mozambique Current. Major drift sequences can first be identified during the Early Miocene. Consistent with earlier findings, a progressive northward shift of the depocenter indicates that, on a geological timescale, a steady but variable Mozambique Current existed from this time onward. It can furthermore be shown that, during the Early/Middle Miocene, a coast-parallel current was established off the Limpopo River as part of a lee eddy system driven by the Mozambique Current. Modern sedimentation is controlled by the interplay between slope morphology and the lee eddy system, resulting in upwelling of Antarctic Intermediate Water. Drift accumulations at larger depths are related to the reworking of sediment by deep-reaching eddies that migrate southward, forming the Mozambique Current and eventually merging with the Agulhas Current.

Conflict and cognitive control during sentence comprehension: recruitment of a frontal network during the processing of Spanish object-first sentences.

During sentence processing there is a preference to treat the first noun phrase found as the subject and agent, unless marked the other way. This preference would lead to a conflict in thematic role assignment when the syntactic structure conforms to a non-canonical object-before-subject pattern. Left perisylvian and fronto-parietal brain networks have been found to be engaged by increased computational demands during sentence comprehension, while event-reated brain potentials have been used to study the on-line manifestation of these demands. However, evidence regarding the spatiotemporal organization of brain networks in this domain is scarce. In the current study we used Magnetoencephalography to track spatio-temporally brain activity while Spanish speakers were reading subject- and object-first cleft sentences. Both kinds of sentences remained ambiguous between a subject-first or an object-first interpretation up to the appearance of the second argument. Results show the time-modulation of a frontal network at the disambiguation point of object-first sentences. Moreover, the time windows where these effects took place have been previously related to thematic role integration (300–500 ms) and to sentence reanalysis and resolution of conflicts during processing (beyond 500 ms post-stimulus). These results point to frontal cognitive control as a putative key mechanism which may operate when a revision of the sentence structure and meaning is necessary

Technical developments for computed tomography on the CENBG nanobeam line

The use of ion microbeams as probes for computedtomography has proven to be a powerful tool for the three-dimensional characterization of specimens a few tens of micrometers in size. Compared to other types of probes, the main advantage is that quantitative information about mass density and composition can be obtained directly, using specific reconstruction codes. At the Centre d’Etudes Nucléaires de Bordeaux Gradignan (CENBG), this technique was initially developed for applications in cellular biology. However, the observation of the cell ultrastructure requires a sub-micron resolution. The construction of the nanobeamline at the Applications Interdisciplinaires des Faisceaux d’Ions en Region Aquitaine (AIFIRA) irradiation facility has opened new perspectives for such applications. The implementation of computedtomography on the nanobeamline of CENBG has required a careful design of the analysis chamber, especially microscopes for precise sample visualization, and detectors for scanning transmission ion microscopy (STIM) and for particle induced X-ray emission (PIXE). The sample can be precisely positioned in the three directions X, Y, Z and a stepper motor coupled to a goniometer ensures the rotational motion. First images of 3D tomography were obtained on a reference sample containing microspheres of certified diameter, showing the good stability of the beam and the sample stage, and the precision of the motion.

Las presentaciones de trabajo en lengua inglesa: una propuesta de curso on-line.

Desde la explosión de crecimiento de internet que comenzó en los años 90, se han ido creando y poniendo a disposición de los usuarios diversas herramientas para compartir información y servicios de diversas formas, desde el nacimiento del primer navegador hasta nuestros días, donde hay infinidad de lenguajes aplicables al ámbito web. En esta fase de crecimiento, en primer lugar, de cara a usuarios individuales, saldrían herramientas que permitirían a cada cual hacer su web personal, con sus contenidos expuestos. Más adelante se fue generando el fenómeno “comunidad”, con, por ejemplo, foros, o webs en las que había múltiples usuarios que disfrutaban de contenidos o servicios que la web ofreciese. Este crecimiento del mundo web en lo comunitario ha avanzado en muchas ramas,entre ellas, por supuesto, la educacional, surgiendo plataformas como la que es base del proyecto que a continuación se presenta, y herramienta básica y prácticamente ya imprescindible en la enseñanza universitaria: Moodle. Moodle es una herramienta diseñada para compatir recursos y diseñar actividades para el usuario potencial, complementando su aprendizaje en aula, o incluso siendo una vía autónoma de aprendizaje en sí misma. Se ha realizado un estudio sobre el estado de saludo de los contenidos que se exponen en Moodle, y se ha encontrado que una gran mayoría de los cursos que se pueden visitar tienen un gran número de carencias. Por un lado, hay pocos con material original explotado exclusivamente para el curso, y, si tienen material original, no se ha observado una especial atención por la maquetación. Por otro lado, hay muchos otros sin material original, y, en ambos casos, no se ha encontrado ningún curso que ofrezca material audiovisual exclusivo para el curso, presentando algunos en su lugar material audiovisual encontrado en la red (Youtube, etc). A la vista de estos hechos, se ha realizado un proyecto que intenta aportar soluciones ante estas carencias, y se presenta un curso procedente de diversas referencias bibliográficas, para la parte textual, y material audiovisual original e inédito que también se ha explotado específicamente para este curso. Este material ha sido por un lado vídeo, que se ha visionado, editado y subtitulado con software de libre distribución, y por otro lado, audio, que complementa un completo glosario que se ha añadido como extra al curso y cuyo planteamiento no se ha encontrado en ningún curso online de los revisados. Todo esto se ha envuelto en una maquetación cuidada que ha sido fruto del estudio de los lenguajes web html y CSS, de forma que, por un lado, el curso sea un lugar agradable en el que aprender dentro de internet, y por otro, se pudiesen realizar ciertas operaciones que sin estos conocimientos habrían sido imposibles, como la realización del glosario o la incrustación de imágenes y vídeos. A su vez, se ha tratado de dar un enfoque didáctico a toda la memoria del proyecto, de forma que pueda ser de utilidad a un usuario futuro que quisiese profundizar en los usos de Moodle, introducirse en el lenguaje web, o introducirse en el mundo de la edición de vídeo. ABSTRACT: Since the explosion of Internet growth beginning in the 90s, many tools have been created and made available for users to share information and services in various ways, from the birth of the first browser until today, where there are plenty of web programming languages. This growth stage would give individual users tools that would allow everyone to make an own personal website, with their contents exposed. Later, the "community" phenomenon appeared with, for example, forums, or websites where multiple users enjoyed the content or web services that those websites offered. Also, this growth in the web community world has progressed in many fields, including education, with the emerge of platforms such as the one that this project uses as its basis, and which is the basic and imperative tool in college education: Moodle. Moodle is a tool designed to share resources and design activities for the potential user, completing class learning, or even letting this user learn in an autonomous way. In this project a study on the current situation of the content present in Moodle courses around the net has been carried out, and it has been found that most of them lack of original material exploited exclusively for the courses, and if they have original material, there has been not observed concern on the layout where that material lies. On the other hand, there are many other with non original material, and in both cases, there has not been found any course that offers audio- visual material made specifically for the course, instead of presenting some audiovisual material found on the net (Youtube, etc). In view of these facts, the project presented here seeks to provide solutions to these shortcomings, presenting a course with original material exploited from various references, and unpublished audioevisual material which also has been exploited specifically for this course. This material is, on one hand, video, which has been viewed, edited and subtitled with free software, and on the other, audio, which complements a comprehensive glossary that has been added as an extra feature to the course and whose approach was not found in any of the online courses reviewed. All of this has been packaged in a neat layout that has been the result of the study of web languages HTML and CSS, so that first, the course was a pleasant place to learn on the internet, and second, certain operations could be performed which without this knowledge would have been impossible, as the glossary design or embedding images and videos. Furthermore, a didactic approach has been adopted to the entire project memory, so it can be useful to a future user who wanted to go deeper on the uses of Moodle, containing an intro into the web language, or in the world video editing.

LED backlight designs with the flow-line method

An LED backlight has been designed using the flow-line design method. This method allows a very efficient control of the light extraction. The light is confined inside the guide by total internal reflection, being extracted only by specially calculated surfaces: the ejectors. Backlight designs presented here have a total optical efficiency of up to 80% (including Fresnel and absorption losses) with an FWHM below 30 degrees. The experimental results of the first prototype are shown.

Characterization of a Right Angle Transfer Point in a Fruit Packing Line

Characteristics of the impacts su!ered by the fruit on a transfer point of an experimental fruit packing line were analysed. The transfer is made up by two transporting belts at di!erent heights forming an angle of 903. These transfer points are very common in fruit packing lines, in which fruits receive two impacts: the "rst onto the belt base and the second into the lateral plate. Diferent tests were carried out to study the e!ect of transfer height, velocity, belt structure and padding on the acceleration values recorded by an instrumental sphere (IS 100). Results showed that transfer height and belt structure a!ect mainly impact values on the belt base, and padding a!ects mainly impact values registered for lateral contact. The elect of belt velocity in both impacts is less important when compared to the rest of the variables. Additionally, two powered transfer decelerators were tested at the same point with the aim of decreasing impacts su!ered by the fruit. Comparing impacts registered using these decelerators to those analysed in the first part of the study without decelerators, a high reduction of the impact values was observed.

Characterization of a 90° Transfer Point in a Fruit Packing Line.

Characteristics of the impacts suffered by the fruit on a transfer point of an experimental fruit packing line were analysed. The transfer is made up by two transporting belts at different heights forming an angle of 90°. These transfer points are very common in fruit packing lines, in which fruits receive two impacts: the first onto the belt base and the second into the lateral plate. Different tests were earned out to study the effect of transfer height, velocity, belt structure and padding on tire acceleration values recorded by an instrumental sphere (IS 100). Results showed that transfer height and belt structure affect mainly impact values on the belt base, and padding affects mainly impact values registered in lateral impact. The effect of belt velocity in both impacts is less important when compared to the rest of the variables. Additionally, two powered transfer decelerators were tested at the same point with the aim of decreasing impacts suffered by the fruit. Comparing impacts registered using these decelerators to those analysed in the first part of the study without decelerators a high reduction of the impact values was observed.

Estudio acústico del estadio "La Peineta" utilizando line arrays

El proyecto fin de carrera consiste en un estudio acústico del Estadio de la Peineta (estadio de fútbol perteneciente al club Atlético de Madrid el cual se encuentra en construcción). Se realizará el diseño por completo de una maqueta del estadio utilizando el programa EASE. Esta maqueta se hará a escala real, exactamente como se está construyendo el estadio. A dicha maqueta se le incorporarán los diferentes materiales absorbentes específicos a cada una de las superficies que compongan el estadio. Se crearán tantas zonas de audiencia como superficies se obtenga en cada grada sobre donde se realizarán dos estudios acústicos diferentes. El primer estudio se realizará con un total de 24 clústeres de altavoces los cuales están compuestos por 10 altavoces Aero 50. La colocación estratégica de cada uno de estos altavoces se estudiará con la herramienta EASE Focus 2. Una vez obtenidas las posiciones se importarán cada uno de estos clústeres de altavoces su respectiva configuración. El otro estudio se realizará con la mitad de arrays de altavoces con la intención de tener una comparativa de recubrimiento entre un estudio y otro. Las pruebas de simulación serán analizando el nivel de presión sonora que provoca cada uno de estos estudios cuando tienen sus altavoces en funcionamiento. Se utilizará el módulo de “Area mapping” para estudiar el mapeo en cada una de las zonas de audiencia definidas, así como cada una de las distribuciones por área que predominan. Se irán anotando cada uno de los problemas e ideas que van surgiendo a lo largo de dicho proyecto para proponer una continuación y mejora del mismo. Se planteará una serie de pasos e pruebas al final de la memoria ya que se trata de un proyecto sin cerrar y puede ser continuado por otra persona. ABSTRACT. The final project is an acoustic studio Peineta Stadium (soccer stadium belongs to the club Atletico Madrid which is under construction). The design is made entirely of a model of the stadium using the EASE program. This model will be full scale, exactly as is building the stadium. A model that should be incorporated into the various specific absorbent material to each of the surfaces that make up the stadium. Hearing so many areas as surfaces is obtained in each tier on which two different acoustic studies will be conducted will be created. The first study was conducted with a total of 24 speaker clusters which are composed of 10 speakers Aero 50. The strategic placement of each of these speakers will be studied with the EASE Focus 2. Once obtained tool positions are imported each these clusters of the respective speaker configuration. The other study was conducted with half speaker arrays with the intention of having a comparative study between a coating and another. Simulation tests will be analyzing the sound pressure level which causes each of these studies have their speakers when in operation. Module "mapping area" will be used to study the mapping in each of the areas defined audience, and each of the area distributions predominate. They will be written down each of the issues and ideas that arise throughout the project to propose a continuation and improvement. a series of steps and tests at the end of the memory will be raised because it is a project without closing and may be continued for another person.