TURNOVER RATE OF THE NEURONAL CONNEXIN CX36 IN HELA CELLS

Electrical synapses formed of the gap junction protein Cx36 show a great deal of functional plasticity, much dependent on changes in phosphorylation state of the connexin. However, gap junction turnover may also be important for regulating cell-cell communication, and turnover rates of Cx36 have not been studied. Connexins have relatively fast turnover rates, with short half-lives measured to be 1.5 to 3.5 hours in pulse-chase analyses of connexins (Cx26 and Cx43) in tissue culture cells and whole organs. We utilized HaloTag technology to study the turnover rate of Cx36 in transiently transfected HeLa cells. The HaloTag protein forms irreversible covalent bonds with chloroalkane ligands, allowing pulse-chase experiments to be performed very specifically. The HaloTag open reading frame was inserted into an internal site in the C-terminus of Cx36 designed not to disrupt the regulatory phosphorylation sites and not to block the C-terminal PDZ interaction motif. Functional properties of Cx36-Halo were assessed by Neurobiotin tracer coupling, live cell imaging, and immunostaining. For the pulse-chase study, transiently transfected HeLa cells were pulse labeled with Oregon Green (OG) HaloTag ligand and chase labeled at various times with tetramethylrhodamine (TMR) HaloTag ligand. Cx36-Halo formed large junctional plaques at sites of contact between transfected HeLa cells and was also contained in a large number of intracellular vesicles. The Cx36-Halo transfected HeLa cells supported Neurobiotin tracer coupling that was regulated by activation and inhibition of PKA in the same manner as wild-type Cx36 transfected cells. In the pulse-chase study, junctional protein labeled with the pulse ligand (OG) was gradually replaced by newly synthesized Cx36 labeled with the chase ligand (TMR). The half-life for turnover of protein in junctional plaques was 2.8 hours. Treatment of the pulse-labeled cells with Brefeldin A (BFA) prevented the addition of new connexins to junctional plaques, suggesting that the assembly of Cx36 into gap junctions involves the traditional ER-Golgi-TGN-plasma membrane pathway. In conclusion, Cx36-Halo is functional and has a turnover rate in HeLa cells similar to that of other connexins that have been studied. This turnover rate is likely too slow to contribute substantially to short-term changes in coupling of neurons driven by transmitters such as dopamine, which take minutes to achieve. However, turnover may contribute to longer-term changes in coupling.

Investigation of the mechanism of increased melanoma incidence in UV -irradiated mouse skin

The availability of transplantable, syngeneic murine melanomas made it possible to study the potential effects of UV radiation on the growth and progression of melanomas in an animal model. The purpose of my study was to determine how UV-irradiation increases the incidence of melanoma out-growth, when syngeneic melanoma cells are transplanted into a UV-irradiated site. Short term intermittent UVB exposure produces a transitory change in the mice which allows the increased outgrowth of melanoma cells injected into the UV-irradiated site. One possible mechanism is an immunomodulatory effect of UVR on the host. An alternative mechanism to account for the increased tumor incidence in the UV-irradiated site, is the release of inflammatory mediators from UV-irradiated epidermal cells. A third possibility is that UVR could induce the production and/or release of melanoma-specific growth factors resulting in increased melanoma outgrowth.^ My first step in distinguishing among these different possible mechanisms was to characterize further the conditions leading to increased development of melanoma cells in UV-irradiated mouse skin. Next, I attempted to determine which of the 3 proposed mechanisms was most likely. To do this, I defined the specificity of the effect by examining the growth of additional C3H tumorigenic cell lines in UV-irradiated skin. Second, I determined the immunogenicity of these tumor cell lines. The tumor cell lines exhibiting increased tumor incidence are restricted to those tumor cell lines which are immunogenic in normal C3H mice. Third, I determined the effect of UVR on melanoma development did not occur in immunosuppressed mice.^ Because of results from these three lines of investigation suggested that the effect was immunologically mediated, I then investigated whether specific immune reactions were affected by local UV irradiation. To accomplish this, I investigated the effect of UVR on cutaneous immune cells and on induction of contact hypersensitivity (CHS), and I also determined the effect of UVR on the development and the expression of systemic immunity against the melanoma cells. There is no clear cut relationship between the number of Langerhans or Thy1+ cells and the UV effect on tumor incidence. Furthermore, there was no suppression of CHS in the UV-irradiated mice. While the development of systemic immunity is significantly reduced, it appears to be sufficient to provide in vivo immunity to tumor challenge. However the elicitation of tumor immunity in immunized mice can be abrogated if tumor challenge occurs in the site of UV irradiation. This investigation provides new information on an effect of UVR on the elicitation of tumor immunity. Furthermore, it indicates that UV radiation can play a role in the development of melanoma other than just in the transformation of melanocytes. ^

Functional analysis of p53 phosphorylation and a p53 hot spot mutation

The tumor suppressor p53 is a phosphoprotein which functions as a transcriptional activator. By monitoring the transcriptional activity, we studied how p53 functions is regulated in relation to cell growth and contact inhibition. When cells were arrested at G1 phase of the cell cycle by contact inhibition, we found that p53 transactivation function was suppressed. When contact inhibition was overridden by cyclin E overexpression which stimulates cell cycle progression, p53 function was restored. This observation led to the development of a cell density assay to study the regulation of p53 function during cell cycle for the functional significance of p53 phosphorylation. The murine p53 is phosphorylated at serines 7, 9, 12, 18, 37, 312 and 389. To understand the role of p53 phosphorylation, we generated p53 constructs encoding serine-to-alanine or serine-to-glutamate mutations at these codons. The transcriptional activity were measured in cells capable of contact inhibition. In low-density cycling cells, no difference in transcriptional activity was found between wild type p53 and any of the mutants. In contact-inhibited cells, however, only mutations of p53 at serine 389 resulted in altered responses to cell cycle arrest and to cyclin E overexpression. The mutant with serine-to-glutamate substitution at codon 389 retained its function in contact inhibited cells. Cyclin E overexpression in these cells induced p53 phosphorylation at serine 389. Furthermore, we showed that phosphorylation at serine 389 regulates p53 DNA binding activity. Our findings implicate that phosphorylation is an important mechanism for p53 activation.^ p53 is the most frequently mutated gene in human tumors. To study the mechanism of p53 inactivation by mutations, we carried out detailed analysis of a murine p53 mutation with an arginine-to-tryptophane substitution at codon 245. The corresponding human p53 mutation at amino acid 248 is the most frequently mutated codon in tumors. We showed that this mutant is inactive in suppressing focus formation, binding to DNA and transactivation. Structural analysis revealed that this mutant assumes the wild type protein conformation. These findings define a novel class of p53 mutations and help to understand structure-function relationship of p53. ^

NMR structural analysis of cardiac troponin C: Monitoring conformational changes induced by binding calcium, troponin I, and calcium -sensitizing drugs

Contraction of cardiac muscle is regulated through the Ca2+ dependent protein-protein interactions of the troponin complex (Tn). The critical role cardiac troponin C (cTnC) plays as the Ca2+ receptor in this complex makes it an attractive target for positive inotropic compounds. In this study, the ten Met methyl groups in cTnC, [98% 13C ϵ]-Met cTnC, are used as structural markers to monitor conformational changes in cTnC and identify sites of interaction between cTnC and cardiac troponin I (cTnI) responsible for the Ca2+ dependent interactions. In addition the structural consequences that a number of Ca2+-sensitizing compounds have on free cTnC and the cTnC·cTnI complex were characterized. Using heteronuclear NMR experiments and monitoring chemical shift changes in the ten Met methyl 1H-13C correlations in 3Ca2+ cTnC when bound to cTnI revealed an anti-parallel arrangement for the two proteins such that the N-domain of cTnI interacts with the C-domain of cTnC. The large chemical shifts in Mets-81, -120, and -157 identified points of contact between the proteins that include the C-domain hydrophobic surface in cTnC and the A, B, and D helical interface located in the regulatory N-domain of cTnC. TnI association [cTnI(33–80), cTnI(86–211), or cTnI(33–211)] was found also to dramatically reduce flexibility in the D/E central linker of cTnC as monitored by line broadening in the Met 1H- 13C correlations of cTnC induced by a nitroxide spin label, MTSSL, covalently attached to cTnC at Cys 84. TnI association resulted in an extended cTnC that is unlike the compact structure observed for free cTnC. The Met 1H-13C correlations also allowed the binding characteristics of bepridil, TFP, levosimendan, and EMD 57033 to the apo, 2Ca2+, and Ca2+ saturated forms of cTnC to be determined. In addition, the location of drug binding on the 3Ca2+cTnC·cTnI complex was identified for bepridil and TFP. Use of a novel spin-labeled phenothiazine, and detection of isotope filtered NOEs, allowed identification of drug binding sites in the shallow hydrophobic cup in the C-terminal domain, and on two hydrophobic surfaces on N-regulatory domain in free 3Ca2+ cTnC. In contrast, only one N-domain drug binding site exists in 3Ca2+ cTnC·cTnI complex. The methyl groups of Met 45, 60 and 80, which are grouped in a hydrophobic patch near site II in cTnC, showed the greatest change upon titration with bepridil or TFP, suggesting that this is a critical site of drug binding in both free cTnC and when associated with cTnI. The strongest NOEs were seen for Met-60 and -80, which are located on helices C and D, respectively, of Ca2+ binding site II. These results support the conclusion that the small hydrophobic patch which includes Met-45, -60, and -80 constitutes a drug binding site, and that binding drugs to this site will lead to an increase in Ca2+ binding affinity of site II while preserving maximal cTnC activity. Thus, the subregion in cTnC makes a likely target against which to design new and selective Ca2+-sensitizing compounds. ^

Characterization of diagenetic fluids at ODP Site 103-639

Site 639, drilled during Leg 103 of the Ocean Drilling Program, penetrated an Upper Jurassic to Lower Cretaceous carbonate platform on a tilted fault block along the Galicia margin off the northwest Iberian Peninsula. The carbonate platform is composed primarily of a sequence of dolomite overlying limestone. Samples were analyzed for mineral chemistry, stable isotope geochemistry, fluid inclusion microthermometry, and volatile contents and by dolomite pyrolysis mass spectrometry for trace sulfate minerals. The dolomite recovered from the Galicia margin at Site 639 formed during shallow burial from sulfate-bearing, hypersaline brines at slightly elevated temperatures. The light oxygen isotopic signatures of the dolomite are interpreted as the result of the evaporative loop and slightly elevated temperatures during dolomite formation or from reequilibration at higher temperatures during deeper burial. The hypersalinity is interpreted to be associated with a nearby, shallow restricted basin that formed during rifting of the Iberian margin from Newfoundland. The dolomitization of the platform is therefore a by-product of the rifting.

Macroturbidites of ODP Hole 103-641C

Barremian through uppermost Aptian strata from ODP Hole 641C, located upslope of a tilted fault block on the Galicia margin (northwest Spain), are syn-rift sediments deposited in the bathyal realm and are characterized by rapid sedimentation from turbidity currents and debris flows. Calcarenite and calcirudite turbidites contain shallow-water carbonate, terrigenous, and pelagic debris, in complete or partial Bouma sequences. These deposits contain abraded micritized bioclasts of reefal debris, including rudist fragments. The youngest turbidite containing shallow-water carbonate debris at Site 641 defines the boundary between syn-rift and post-rift sediments; this is also the boundary between Aptian and Albian sediments. Some Aptian turbidites are partially silicified, with pore-filling chalcedony and megaquartz. Adjacent layers of length-fast and -slow chalcedony are succeeded by megaquartz as the final pore-filling stage within carbonate reef debris. Temperatures of formation, calculated from the oxygen isotopic composition of the authigenic quartz, are relatively low for formation of quartz but are relatively warm for shallow burial depths. This quartz cement may be interpreted as a rift-associated precipitate from seawater-derived epithermal fluids that migrated along a fault associated with the tilted block and were injected into the porous turbidite beds. These warm fluids may have cooled rapidly and precipitated silica at the boundaries of the turbidite beds as a result of contact with cooler pore waters. The color pattern in the quartz cement, observed by cathodoluminescence and fluorescence techniques, and changes in the trace lement geochemistry mimic the textural change of the different quartz layers and indicates growth synchronism of the different quartz phases. Fluorescence petrography of neomorphosed low-Mg-calcite bioclasts in the silicified turbidites shows extensive zonation and details of replacive crystal growth in the bioclasts that are not observed by cathodoluminescence. Fluorescence microscopy also reveals a competitive growth history during neomorphism of the adjacent crystals in an altered carbonate bioclast. Barremian-Aptian background pelagic sediments from Hole 641C have characteristics similar to pelagic sediments from the Blake-Bahama Formation described by Jansa et al. (1979) from the western North Atlantic. Sediments at this site differ from the Blake-Bahama Formation type locality in that the Barremian-Aptian pelagic sediments have a higher percentage of dark calcareous claystone and some turbidites are silicified at Site 641. The stable isotopic composition of the pelagic marlstones from Site 641 is similar to those of other Berriasian-Aptian pelagic sediments from the Atlantic.

Contents of P2O5 and REE in phosphorites from shelves and Pacific seamounts

Recent phosphorites from the Namibian shelf are characterized by low REE contents, depletion in REE compared to host sediments and sharp deficiency of lanthanum and europium. In Late Quaternary and Pre-Quaternary phosphorites from ocean shelves REE contents and patterns in general are the same as in host sediments. Phosphorites from seamounts are enriched in REE compared to shelf phosphorites and their patterns are close to one of seawater. Behavior of REE in shelf phosphorites is determined by the fact that in early stages of phosphorite formation REE are associated not primarily with phosphate, but with organic matter and terrigenous impurities. Only in the later stages of diagenesis phosphate begins to play a leading role in concentration of REE. In metasomatic phosphorites on seamounts concentration of REE depends on age and depth of these rocks, i.e. it is determined by duration and conditions of contact with sea water.

Thecosomata in late Quternary deposits of ODP Holes 134-830A and 134-831A

We undertook a quantitative study of Thecosomata shells (pelagic gastropods) and their remains in Quaternary foraminiferal oozes deposited on the tilted calcareous platform of the Bougainville Guyot (Hole 831 A), and in the late Quaternary volcanic siltstones, claystones and sandy interbeds on the upper forearc slope of the central New Hebrides Island Arc (Hole 830A). The distribution of the species is based on the identification of adult shells, juvenile stages, protoconchs, and characteristic shell fragments. By studying thecosomatous shells using a scanning electron microscope (SEM), we were able to specify the fine microstructure of the coiled Limacina inflata and compare it with the rod-type crossed-lamellar structure of some other Limacina species, as well as with the helical structure of the Cavoliniidae.

Dinoflagellate cysts of ODP Leg 101 holes

The Albian-Cenomanian sediments in Holes 627B and 635B contain diverse dinoflagellate-cyst assemblages, which show affinities with coeval assemblages from offshore Morocco and northwest Europe. A total of 34 samples were analyzed from the shallow-water platform sediments and neritic marly chalk of Hole 627B and from the argillaceous chalk and limestone of Hole 635B. Dinoflagellate cysts indicate that the top of the shallow-water platform drilled at Hole 627B must be attributed to the late Albian. Dinocysts also date the drowning of the carbonate platform of the Blake Plateau. This drowning started in the latest Albian (Vraconian) and continued into the Cenomanian. The site area changed from an inner to intermediate or outer(?) neritic environment. The area around Hole 635B from the late Albian appears to have been situated in a deeper environment than the area around Hole 627B during the same period. The new dinoflagellate-cyst species Compositosphaeridiuml bahamaensis n. sp., Maghrebinia breviornata n. sp., and Subtilisphaeral habibi n. sp. are described, and Pervosphaeridium truncatum is emended. Additional taxonomic remarks about other species are included.

(Table 1) Chemical and isotopic compositions of rudist aragonite and magnesian calcite cements from ODP Hole 144-877A

The classic paleotemperature record based on d18O data from pelagic foraminiferal calcite suggests that equatorial sea-surface temperatures during the Maastrichtian (~12-20°C) were much cooler than today (~27-29°C). Such cool equatorial temperatures contradict basic theories of tropical atmospheric and ocean dynamics. We report d18O data from remarkably well preserved rudist aragonite and magnesian calcite cements of Maastrichtian age (~69+/-1 Ma) from the carbonate platform of Wodejebato guyot in the western Pacific. These data suggest that equatorial sea-surface temperatures in the Maastrichtian (best estimate ~27-32°C) were at least as warm as today. This finding helps reconcile the geologic d18O record with ocean-atmospheric dynamic theory and implies a reduction in the poleward heat flux required by global climate simulations of greenhouse conditions.

Prediction of bendig load capacity of timber beams by finite element method simulation of knots and grain deviation

A finite element model was used to simulate timberbeams with defects and predict their maximum load in bending. Taking into account the elastoplastic constitutive law of timber, the prediction of fracture load gives information about the mechanisms of timber failure, particularly with regard to the influence of knots, and their local graindeviation, on the fracture. A finite element model was constructed using the ANSYS element Plane42 in a plane stress 2D-analysis, which equates thickness to the width of the section to create a mesh which is as uniform as possible. Three sub-models reproduced the bending test according to UNE EN 408: i) timber with holes caused by knots; ii) timber with adherent knots which have structural continuity with the rest of the beam material; iii) timber with knots but with only partial contact between knot and beam which was artificially simulated by means of contact springs between the two materials. The model was validated using ten 45 × 145 × 3000 mm beams of Pinus sylvestris L. which presented knots and graindeviation. The fracture stress data obtained was compared with the results of numerical simulations, resulting in an adjustment error less of than 9.7%

Bioaccessibility of heavy metals and risk assessment in urban gardens of Madrid

Human health problems and solutions. Urban gardening has spread worldwide in recent years as it enhances food security and selfsupply and promotes community integration. However urban soils are significantly enriched in trace elements relative to background levels. Exposure to the soil in urban gardens may therefore result in adverse health effects depending on the degree of contact during gardening, infant recreational activities and ingestion of vegetables grown in them. In order to evaluate this potential risk, 36 composite samples were collected from the top 20 cm of the soil of 6 urban gardens in Madrid. The aqua regia (pseudototal) and glycine-extractable (bioaccessible) concentrations of Co, Cr, Cu, Ni, Pb and Zn were determined by atomic absorption spectrophotometry. Additionally, pH, texture, Fe, Ca, and Mn concentrations, and organic matter and calcium carbonate contents were determined in all urban gardens and their influence on trace element bioaccessibility was analyzed.

Diseño y configuración de una red de comunicaciones en un Centro de Contactos - Contact Center

Este proyecto muestra una solución de red para una empresa que presta servicios de Contact Center desde distintas sedes distribuidas geográficamente, utilizando la tecnología de telefonía sobre IP. El objetivo de este proyecto es el de convertirse en una guía de diseño para el despliegue de soluciones de red utilizando los actuales equipos de comunicaciones desarrollados por el fabricante Cisco Systems, Inc., los equipos de seguridad desarrollados por el fabricante Fortinet y los sistemas de telefonía desarrollados por Avaya Inc. y Oracle Corporation, debido a su gran penetración en el mercado y a las aportaciones que cada uno ha realizado en el sector de Contact Center. Para poder proveer interconexión entre las sedes de un Contact Center se procede a la contratación de un acceso a la red MPLS perteneciente a un operador de telecomunicaciones, quien provee conectividad entre las sedes utilizando la tecnología VPN MPLS con dos accesos diversificados entre sí desde cada una de las sedes del Contact Center. El resultado de esta contratación es el aprovechamiento de las ventajas que un operador de telecomunicaciones puede ofrecer a sus clientes, en relación a calidad de servicio, disponibilidad y expansión geográfica. De la misma manera, se definen una serie de criterios o niveles de servicio que aseguran a un Contact Center una comunicación de calidad entre sus sedes, entendiéndose por comunicación de calidad aquella que sea capaz de transmitirse con unos valores mínimos de pérdida de paquetes así como retraso en la transmisión, y una velocidad acorde a la demanda de los servicios de voz y datos. Como parte de la solución, se diseña una conexión redundante a Internet que proporciona acceso a todas las sedes del Contact Center. La solución de conectividad local en cada una de las sedes de un Contact Center se ha diseñado de manera general acorde al volumen de puestos de usuarios y escalabilidad que pueda tener cada una de las sedes. De esta manera se muestran varias opciones asociadas al equipamiento actual que ofrece el fabricante Cisco Systems, Inc.. Como parte de la solución se han definido los criterios de calidad para la elección de los Centros de Datos (Data Center). Un Contact Center tiene conexiones hacia o desde las empresas cliente a las que da servicio y provee de acceso a la red a sus tele-trabajadores. Este requerimiento junto con el acceso y servicios publicados en Internet necesita una infraestructura de seguridad. Este hecho da lugar al diseño de una solución que unifica todas las conexiones bajo una única infraestructura, dividiendo de manera lógica o virtual cada uno de los servicios. De la misma manera, se ha definido la utilización de protocolos como 802.1X para evitar accesos no autorizados a la red del Contact Center. La solución de voz elegida es heterogénea y capaz de soportar los protocolos de señalización más conocidos (SIP y H.323). De esta manera se busca tener la máxima flexibilidad para establecer enlaces de voz sobre IP (Trunk IP) con proveedores y clientes. Esto se logra gracias a la utilización de SBCs y a una infraestructura interna de voz basada en el fabricante Avaya Inc. Los sistemas de VoIP en un Contact Center son los elementos clave para poder realizar la prestación del servicio; por esta razón se elige una solución redundada bajo un entorno virtual. Esta solución permite desplegar el sistema de VoIP desde cualquiera de los Data Center del Contact Center. La solución llevada a cabo en este proyecto está principalmente basada en mi experiencia laboral adquirida durante los últimos siete años en el departamento de comunicaciones de una empresa de Contact Center. He tenido en cuenta los principales requerimientos que exigen hoy en día la mayor parte de empresas que desean contratar un servicio de Contact Center. Este proyecto está dividido en cuatro capítulos. El primer capítulo es una introducción donde se explican los principales escenarios de negocio y áreas técnicas necesarias para la prestación de servicios de Contact Center. El segundo capítulo describe de manera resumida, las principales tecnologías y protocolos que serán utilizados para llevar a cabo el diseño de la solución técnica de creación de una red de comunicaciones para una empresa de Contact Center. En el tercer capítulo se expone la solución técnica necesaria para permitir que una empresa de Contact Center preste sus servicios desde distintas ubicaciones distribuidas geográficamente, utilizando dos Data Centers donde se centralizan las aplicaciones de voz y datos. Finalmente, en el cuarto capítulo se presentan las conclusiones obtenidas tras la elaboración de la presente memoria, así como una propuesta de trabajos futuros, que permitirían junto con el proyecto actual, realizar una solución técnica completa incluyendo otras áreas tecnológicas necesarias en una empresa de Contact Center. Todas las ilustraciones y tablas de este proyecto son de elaboración propia a partir de mi experiencia profesional y de la información obtenida en diversos formatos de la bibliografía consultada, excepto en los casos en los que la fuente es mencionada. ABSTRACT This project shows a network solution for a company that provides Contact Center services from different locations geographically distributed, using the Telephone over Internet Protocol (ToIP) technology. The goal of this project is to become a design guide for performing network solutions using current communications equipment developed by the manufacturer Cisco Systems, Inc., firewalls developed by the manufacturer Fortinet and telephone systems developed by Avaya Inc. and Oracle Corporation, due to their great market reputation and their contributions that each one has made in the field of Contact Center. In order to provide interconnection between its different sites, the Contact Center needs to hire the services of a telecommunications’ operator, who will use the VPN MPLS technology, with two diversified access from each Contact Center’s site. The result of this hiring is the advantage of the benefits that a telecommunications operator can offer to its customers, regarding quality of service, availability and geographical expansion. Likewise, Service Level Agreement (SLA) has to be defined to ensure the Contact Center quality communication between their sites. A quality communication is understood as a communication that is capable of being transmitted with minimum values of packet loss and transmission delays, and a speed according to the demand for its voice and data services. As part of the solution, a redundant Internet connection has to be designed to provide access to every Contact Center’s site. The local connectivity solution in each of the Contact Center’s sites has to be designed according to its volume of users and scalability that each one may have. Thereby, the manufacturer Cisco Systems, Inc. offers several options associated with the current equipment. As part of the solution, quality criteria are being defined for the choice of the Data Centers. A Contact Center has connections to/from the client companies that provide network access to teleworkers. This requires along the access and services published on the Internet, needs a security infrastructure. Therefore is been created a solution design that unifies all connections under a single infrastructure, dividing each services in a virtual way. Likewise, is been defined the use of protocols, such as 802.1X, to prevent unauthorized access to the Contact Center’s network. The voice solution chosen is heterogeneous and capable of supporting best-known signaling protocols (SIP and H.323) in order to have maximum flexibility to establish links of Voice over IP (IP Trunk) with suppliers and clients. This can be achieved through the use of SBC and an internal voice infrastructure based on Avaya Inc. The VoIP systems in a Contact Center are the key elements to be able to provide the service; for this reason a redundant solution under virtual environment is been chosen. This solution allows any of the Data Centers to deploy the VoIP system. The solution carried out in this project is mainly based on my own experience acquired during the past seven years in the communications department of a Contact Center company. I have taken into account the main requirements that most companies request nowadays when they hire a Contact Center service. This project is divided into four chapters. The first chapter is an introduction that explains the main business scenarios and technical areas required to provide Contact Center services. The second chapter describes briefly the key technologies and protocols that will be used to carry out the design of the technical solution for the creation of a communications network in a Contact Center company. The third chapter shows a technical solution required that allows a Contact Center company to provide services from across geographically distributed locations, using two Data Centers where data and voice applications are centralized. Lastly, the fourth chapter includes the conclusions gained after making this project, as well as a future projects proposal, which would allow along the current project, to perform a whole technical solution including other necessary technologic areas in a Contact Center company All illustrations and tables of this project have been made by myself from my professional experience and the information obtained in various formats of the bibliography, except in the cases where the source is indicated.