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FLOW-CYTOMETRIC SORTING OF RAM SPERMATOZOA: PRODUCTION OF LAMBS OF A PRE-DETERMINED SEX USING IN VIVO AND IN VITRO FERTILISATIONHollinshead, Fiona Kate January 2003 (has links)
Abstract Birth of offspring of a pre-determined sex using flow cytometrically sorted fresh spermatozoa was first achieved in rabbits by Johnson et al. (1989). Since then offspring have been produced using sex-sorted spermatozoa from several different species (reviewed by Johnson, 2000). Initially, efficiency of the sex-sorting technology was poor with only low numbers of spermatozoa sorted per hour. Thus, the offspring derived from flow cytometrically sorted spermatozoa were produced with the use of artificial reproductive technologies (ART) such as in vitro fertilisation (IVF) and culture (IVC), intracytoplasmic sperm injection (ICSI) and deep artificial insemination (AI) which facilitated low dose insemination of potentially compromised spermatozoa. More recently, the development of high-speed sorters (Johnson and Welch, 1999) has facilitated the production of offspring using conventional AI techniques with low dose inseminates (Seidel et al., 1999) and successful cryopreservation of sorted spermatozoa (Schenk et al., 1999; Johnson et al., 2000; Lindsey et al., 2002; Schenk and DeGrofft, 2003). Increased efficiency of sorting bull spermatozoa has evolved through significant instrumentation and biological developments which have enabled the commercialization of the sperm sexing technology in the dairy industry, although conception rates in cows after low dose AI with sexed frozen-thawed spermatozoa are still lower than after standard frozen semen AI (Seidel et al., 1999). Subsequently, over 20 000 calves of pre-determined sex have been produced from commercially available sex-sorted frozen-thawed bull spermatozoa (Seidel, 2003). However, similar developments have not been made in the sheep industry and were examined in this thesis. In this study, successful cryopreservation of sex-sorted ram spermatozoa and production of offspring of the pre-determined sex (X: 94.4 %; Y: 100 %) was achieved after low dose (2-4 x 106 total) insemination using conventional laparoscopic intrauterine (IU) AI. However, the overall pregnancy rate for ewes inseminated with sex-sorted frozen-thawed spermatozoa was low (25 %) compared to ewes inseminated with a commercial dose (140 x 106 total) of non-sorted frozen-thawed spermatozoa (54 %). Cryopreservation has been found to not only reduce the proportion of motile spermatozoa, but cause the remaining spermatozoa to undergo changes that advance membrane maturation thereby shortening their lifespan, especially after in vivo fertilisation (Gillan and Maxwell, 1999). It was found that sorting prior to cryopreservation accelerated the maturation of sperm membranes and after co-incubation with oviducal cells in vitro, sorted frozen-thawed spermatozoa were released more rapidly than non-sorted (control) frozen-thawed spermatozoa. The potentially reduced lifespan of sorted frozen-thawed spermatozoa, and practical constraints on the number of spermatozoa that can be sorted for an insemination dose, makes insemination close to the site of fertilisation and time of ovulation critical for successful fertilisation. After treatment of ewes with GnRH to increase the precision of insemination in respect to the time of ovulation, there was no difference in pregnancy rate between ewes inseminated before, during or after the assumed time of ovulation. Furthermore, there was no difference in pregnancy rate after IU AI with similar doses of sorted frozen-thawed and non-sorted frozen-thawed spermatozoa in GnRH-treated ewes. The minimum dose of sorted frozen-thawed spermatozoa required for commercially acceptable pregnancy rates determined after IU AI was high (20 x 106 motile). Consequently, alternative methods for efficiently producing large numbers of offspring of a pre-determined sex using flow cytometrically sorted ram spermatozoa were investigated. Ram spermatozoa can be stored for short periods of time in a chilled state (liquid storage) or for an indefinite period of time in a frozen state (frozen storage; Salamon and Maxwell, 2000). The fixed location of the sperm sorter requires the need for transport of semen from the point of collection to the site of sorting and processing, but also from the sperm sorter site to the recipient females under artificial conditions. In this study, ram spermatozoa liquid stored for 24 h prior to sorting were efficiently sorted, frozen, thawed and after in vitro fertilisation and culture produced a high proportion of grade 1 blastocysts. Similarly, spermatozoa stored at reduced temperatures after sorting maintained high sperm quality for up to 6 days. Furthermore, frozen-thawed spermatozoa from rams and some non-human primates were successfully prepared for sorting and efficiently sorted producing spermatozoa with high quality in vitro parameters. The quality of frozen-thawed ram spermatozoa after sorting was such that successful re-cryopreservation after sorting was possible. Low numbers of frozen-thawed sorted and re-frozen and thawed spermatozoa were optimal for IVF and a high proportion of grade 1 in vitro embryos of a pre-determined sex were produced. These embryos were either transferred immediately or vitrified prior to transfer, extending the application of the sperm sexing technology further. The birth of lambs of pre-determined sex after transfer of both fresh and vitrified embryos derived from frozen-thawed sorted spermatozoa was achieved. The findings in this thesis suggest that sorted frozen-thawed ram spermatozoa may have more advanced membrane maturation state than non-sorted frozen-thawed spermatozoa, resulting in a decreased fertilizing lifespan in the female reproductive tract. Despite this, the use of sexed ram spermatozoa in a number of physiological states (fresh, liquid, frozen) with several different ARTs is possible in producing significant numbers of offspring of a pre-determined sex. Improved efficiency in both sperm sexing and associated reproductive technologies is required for commercialization to be achieved in the sheep industry.
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Psykodynamisk grupphandledning på tre utbildningsnivåer : - erfarenheter av att handleda inom psykoterapi- och handledarutbildningarBernhard, Lisa January 2006 (has links)
Studien är utförd i anslutning till ett pågående forskningsprojekt om grupphandledning i psykoterapi, GUT-projektet. Studiens syfte var att nå ökad kunskap om handledares upplevelse och erfarenhet av att handleda grupper på olika utbildningsnivåer med psykodynamisk inriktning. En kvalitativ ansats valdes för att besvara studiens syfte. Frågeställningarna omfattade handledares tankar om mål och bedömningskriterier för grupphandledning, erfarenheter av tillvägagångssätt i grupphandledning, gruppens inverkan på handledningen samt den organisatoriska ramens betydelse för grupphandledning på olika utbildningsnivåer. Nio handledare ingick i studien. Data har samlats in via öppna semistrukturerade intervjuer och har bearbetats genom tematisk analys. Målen uppgavs skilja sig mellan olika utbildningsnivåer, främst genom olika grad av fördjupning inom till stora delar gemensamma områden. Nivåspecifika skillnader i tillvägagångssätt i handledning redovisades medan respondenternas erfarenheter av gruppens inverkan på handledningen var likartad oavsett utbildningsnivå. Svårigheter förenade med utbildningshandledning i grupp redovisades och den organisatoriska ramens avgörande betydelse för grupphandledningen betonades. Studiens resultat diskuterades i förhållande till befintlig teori.
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Contribution à l'étude du test aléatoire de mémoires RAMFuentes, Antoine David, R.. January 2008 (has links)
Reproduction de : Thèse de docteur-ingénieur : informatique : Grenoble, INPG : 1986. / Titre provenant de l'écran-titre. Bibliogr. p. 139-141.
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FLOW-CYTOMETRIC SORTING OF RAM SPERMATOZOA: PRODUCTION OF LAMBS OF A PRE-DETERMINED SEX USING IN VIVO AND IN VITRO FERTILISATIONHollinshead, Fiona Kate January 2003 (has links)
Abstract Birth of offspring of a pre-determined sex using flow cytometrically sorted fresh spermatozoa was first achieved in rabbits by Johnson et al. (1989). Since then offspring have been produced using sex-sorted spermatozoa from several different species (reviewed by Johnson, 2000). Initially, efficiency of the sex-sorting technology was poor with only low numbers of spermatozoa sorted per hour. Thus, the offspring derived from flow cytometrically sorted spermatozoa were produced with the use of artificial reproductive technologies (ART) such as in vitro fertilisation (IVF) and culture (IVC), intracytoplasmic sperm injection (ICSI) and deep artificial insemination (AI) which facilitated low dose insemination of potentially compromised spermatozoa. More recently, the development of high-speed sorters (Johnson and Welch, 1999) has facilitated the production of offspring using conventional AI techniques with low dose inseminates (Seidel et al., 1999) and successful cryopreservation of sorted spermatozoa (Schenk et al., 1999; Johnson et al., 2000; Lindsey et al., 2002; Schenk and DeGrofft, 2003). Increased efficiency of sorting bull spermatozoa has evolved through significant instrumentation and biological developments which have enabled the commercialization of the sperm sexing technology in the dairy industry, although conception rates in cows after low dose AI with sexed frozen-thawed spermatozoa are still lower than after standard frozen semen AI (Seidel et al., 1999). Subsequently, over 20 000 calves of pre-determined sex have been produced from commercially available sex-sorted frozen-thawed bull spermatozoa (Seidel, 2003). However, similar developments have not been made in the sheep industry and were examined in this thesis. In this study, successful cryopreservation of sex-sorted ram spermatozoa and production of offspring of the pre-determined sex (X: 94.4 %; Y: 100 %) was achieved after low dose (2-4 x 106 total) insemination using conventional laparoscopic intrauterine (IU) AI. However, the overall pregnancy rate for ewes inseminated with sex-sorted frozen-thawed spermatozoa was low (25 %) compared to ewes inseminated with a commercial dose (140 x 106 total) of non-sorted frozen-thawed spermatozoa (54 %). Cryopreservation has been found to not only reduce the proportion of motile spermatozoa, but cause the remaining spermatozoa to undergo changes that advance membrane maturation thereby shortening their lifespan, especially after in vivo fertilisation (Gillan and Maxwell, 1999). It was found that sorting prior to cryopreservation accelerated the maturation of sperm membranes and after co-incubation with oviducal cells in vitro, sorted frozen-thawed spermatozoa were released more rapidly than non-sorted (control) frozen-thawed spermatozoa. The potentially reduced lifespan of sorted frozen-thawed spermatozoa, and practical constraints on the number of spermatozoa that can be sorted for an insemination dose, makes insemination close to the site of fertilisation and time of ovulation critical for successful fertilisation. After treatment of ewes with GnRH to increase the precision of insemination in respect to the time of ovulation, there was no difference in pregnancy rate between ewes inseminated before, during or after the assumed time of ovulation. Furthermore, there was no difference in pregnancy rate after IU AI with similar doses of sorted frozen-thawed and non-sorted frozen-thawed spermatozoa in GnRH-treated ewes. The minimum dose of sorted frozen-thawed spermatozoa required for commercially acceptable pregnancy rates determined after IU AI was high (20 x 106 motile). Consequently, alternative methods for efficiently producing large numbers of offspring of a pre-determined sex using flow cytometrically sorted ram spermatozoa were investigated. Ram spermatozoa can be stored for short periods of time in a chilled state (liquid storage) or for an indefinite period of time in a frozen state (frozen storage; Salamon and Maxwell, 2000). The fixed location of the sperm sorter requires the need for transport of semen from the point of collection to the site of sorting and processing, but also from the sperm sorter site to the recipient females under artificial conditions. In this study, ram spermatozoa liquid stored for 24 h prior to sorting were efficiently sorted, frozen, thawed and after in vitro fertilisation and culture produced a high proportion of grade 1 blastocysts. Similarly, spermatozoa stored at reduced temperatures after sorting maintained high sperm quality for up to 6 days. Furthermore, frozen-thawed spermatozoa from rams and some non-human primates were successfully prepared for sorting and efficiently sorted producing spermatozoa with high quality in vitro parameters. The quality of frozen-thawed ram spermatozoa after sorting was such that successful re-cryopreservation after sorting was possible. Low numbers of frozen-thawed sorted and re-frozen and thawed spermatozoa were optimal for IVF and a high proportion of grade 1 in vitro embryos of a pre-determined sex were produced. These embryos were either transferred immediately or vitrified prior to transfer, extending the application of the sperm sexing technology further. The birth of lambs of pre-determined sex after transfer of both fresh and vitrified embryos derived from frozen-thawed sorted spermatozoa was achieved. The findings in this thesis suggest that sorted frozen-thawed ram spermatozoa may have more advanced membrane maturation state than non-sorted frozen-thawed spermatozoa, resulting in a decreased fertilizing lifespan in the female reproductive tract. Despite this, the use of sexed ram spermatozoa in a number of physiological states (fresh, liquid, frozen) with several different ARTs is possible in producing significant numbers of offspring of a pre-determined sex. Improved efficiency in both sperm sexing and associated reproductive technologies is required for commercialization to be achieved in the sheep industry.
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Anemone an adaptive network memory engine /Hines, Michael R. Gopalan, Kartik. January 2005 (has links)
Thesis (M.S.)--Florida State University, 2005. / Advisor: Dr. Kartik Gopalan, Florida State University, College of Arts and Sciences, Dept. of Computer Science. Title and description from dissertation home page (viewed June 8, 2005). Document formatted into pages; contains ix, 41pages. Includes bibliographical references.
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Análise de objetos a partir da extração da memória RAM de sistemas sobre Android Run-Time (ART) / Objects analisys based on ram memory extraction over android run-time (ART) systemsSoare, Alberto Magno Muniz 16 December 2016 (has links)
Dissertação (mestrado)—Universidade de Brasília, Faculdade de Tecnologia, Departamento de Engenharia Elétrica, 2016. / Submitted by Camila Duarte (camiladias@bce.unb.br) on 2017-01-26T13:34:31Z
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2016_AlbertoMagnoMunizSoares.pdf: 2637603 bytes, checksum: 014fcec35129d64f57837bd31cb0b8b2 (MD5) / Este trabalho tem o objetivo de apresentar uma técnica de análise de objetos em memória no ambiente de execução ART (Android Run-Time) a partir de uma extração de dados da memória volátil. Um estudo do código fonte AOSP (Android Open Source Project) foi feito para entendimento do ambiente de execução utilizado no sistema operacional Android moderno, e foram elaboradas ferramentas de software que permitem a localização, extração e interpretação de dados úteis para o contexto forense. Construídas como extensões para o framework Volatility, essas ferramentas possibilitam localizar, em uma extração de memória de um dispositivo com arquitetura ARM, instâncias de classes arbitrárias e suas propriedades de dados. / The work in this thesis aims at describe a technique for analyzing objects in memory within the execution environment ART (Android Run-Time) from a volatile memory data extraction. A study of the AOSP (Android Open Source Project) source code was necessary to understand the runtime environment used in the modern Android operating system, and software tools were developed allowing the location, extraction and interpretation of useful data for the forensic context. Built as extensions for the Volatility Framework, these tools enable to locate, in a memory extraction from a device with ARM architecture, arbitrary instances of classes and their data properties.
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Modelagem, otimização, construção e avaliação de um protótipo de carneiro hidráulico. / Modeling, optimization and evaluation of a hydraulic ram.Ricardo Nicolas Zarate Rojas 05 December 2002 (has links)
Este trabalho foi realizado no Laboratório de Hidráulica do Departamento de Engenharia Rural da ESALQ/USP, com o objetivo de modelar, otimizar e avaliar um carneiro hidráulico. Para isso foi desenvolvido um modelo teórico, validado no laboratório através da construção de uma eletroválvula comandada eletronicamente por computador. Através da filmagem do jato de água e utilizando método das coordenadas determinou-se a velocidade do jato de água da tubulação de alimentação do carneiro, para três alturas de alimentação e três comprimentos diferentes. Posteriormente foi construído o carneiro hidráulico em PVC, sendo utilizada a eletroválvula como válvula de escape e, por meio de um microprocessador, obtive-se diferentes tempos para o fechamento da válvula, variando entre 0,2 s e 1,6 s. Foram testados dois materiais como tubulação de alimentação, PVC 25mm de diâmetro nominal e aço galvanizado 19,05 mm de diâmetro nominal para três comprimentos e alturas de alimentação. A máxima vazão de recalque obtida foi de 224 L/h, sendo a pressão de recalque de 54 kPa, sendo a tubulação de alimentação PVC, e a altura de alimentação de 2,23 m, para um tempo de pulso da eletroválvula de 0,2 s. O maior rendimento obtido foi 62 % sendo a tubulação de alimentação de PVC, para um tempo de pulso de 0,2 s com uma pressão de 74 kPa e vazão de recalque de 112 L/h. / This work was developed in the Hydraulics Laboratory of the Department of Rural Engineering at ESALQ/USP., with the goal of modeling, optimize and to evaluate a hydraulic ram. For that was developed a theoretical model, validated in the laboratory through the construction of an electronic valve commanded by a computer. Through the filming of water jet and using method of the coordinates tot determined the speed of water jet of the alimentation piping of the ram for three different heights and lengths. It afterwards was built the hydraulic ram in PVC, being used an electronic valve as pulse valve, and by means of the microprocessor Basic Step, was obtained different times to the shutdown of the valve, varying between 0,2 s and 1,6 s. Were tried two material as alimentation piping, PVC 25mm nominal diameter and galvanized steel 19,05 mm nominal diameter for three alimentation lengths. The flow maxim obtained was 224 L/h, being the pressure of emphasizes of 54 kPa, having as alimentation piping PVC, and the height of alimentation of 2,23 m for, for a time of .shutdown of electronic valve of 0,2 s. The biggest obtained revenue was 62%, being the alimentation piping of PVC, for a time of shutdown of 0,2 s with a pressure 74 kPa and a flow of 112 L/h.
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Framtagning av modulbaserad bärande ram till kabinhissar : tillhörande Cibes Lift skruvhissarRahim, Nadir, Wall, Karl Erik January 2017 (has links)
Cibes Lift är en tillverkare av skruvdrivna plattforms-, -kabin och trapphissar. Då intresset har ökat för hissar i privata bostäder vill företaget sänka tillverkningskostnaderna och utöka utbudet av storlekar på deras kabinhissar för att vara konkurrenskraftiga. Syftet med arbetet är att ta fram en ramkonstruktion som ger högre flexibilitet och lättare kan anpassas till flera olika storlekar av kabinhissar. Målet är att konstruera en moduluppbyggd ram med ett maximalt inköpspris på 5500 kronor och uppfylla maskindirektivets säkerhetskrav. För att uppnå arbetets mål analyserades de befintliga hissramarna genom observationer, praktiska lastprov och Finita elementmetoden (FEM). Intervjuer och observationer gjordes hos tillverkarna av ramarna för att ge information om vilka tillverkningsresurser som fanns att tillgå för den nya ramens konstruktion. Med hjälp av idégenereringsmetod skapades därefter två konceptmodeller som utvärderades med utvärderingsmetoden VDI 2225. Utvärderingsmetoden genererade det mest funktionella konceptet som modulerades med CAD-programmet Autodesk Inventor. Optimering utfördes därefter med hjälp av beräkningar för att erhålla en låg tillverkningskostnad samt uppfylla maskindirektivens säkerhetskrav. Resultatet av slutprodukten består av en ram som är uppbyggd av en basmodul och fem stycken variantmoduler som uppfyller samtliga maskindirektivets säkerhetskrav. Ramen är nästintill uteslutande konstruerad av kallvalsade stålprofiler som tillåts att tillverkas i underleverantörens maskinpark. Priset på ramen uppskattas till 4800 kronor vilket avser en basmodul och den största variantmodulen. Priset är baserat på en inköpskvantitet på 500 stycken basmoduler och 100 stycken variantmoduler av vardera storlek per år. I jämförelse med den största befintliga ramen till kabinhissar har vi lyckats sänka priset med 280 %. Slutsatserna av arbetet är att vi har lyckats konstruera en ram som är uppbyggd av moduler och genom användande av standardiserade artiklar har antalet artiklar kunnat sänkas med 58 % i jämförelse med den största befintliga kabinhissen. / Cibes Lift manufacture screw driven stair, platform and cabin Lifts. Among the company´s ambition is to lower their manufacturing costs and expand their cabin model range following a high demand; particularly from the private sector. The purpose of this work is to design a frame easily adaptable to several cabin sizes. The objective is to design a modular frame in accordance with the EU Machinery Directive and with a maximum cost of 5500 SEK. The redesign began with an analysis of the cabin frames, currently in use by the company; in the form of observations, practical load tests and the application of the Finite Element Method (FEM). Interviews and observations were performed at the subcontractor´s facilities, in order to identify production resources availability. Two frame concepts were designed by using a creativity technique and evaluated through the evaluation method VDI2225. Thereafter, using CAD began the creation of the Best Functional Concept; where components were dimensioned using Finite Element Analysis and Strength calculation to meet the objectives; (namely a frame in accordance with the EU Machinery Directive with a maximum cost of 5500 SEK). The result is a modular frame built from one standardized back piece module and five different floor modules; featuring different floor areas. The frame complies with the associated EU Machinery Directives and is almost entirely constructed of cold-formed steel profiles, so manufacturing can be performed using the current subconttor resources. The frame expected cost is of SEK 4800; calculated based on one back piece module plus the largest size floor module. The cost is based upon a purchase quantity of 500 back piece modules and 100 floor modules per year. When comparing this calculated cost to the current frame cost, the estimated price reduction is of 280%. As a conclusion, we have managed to design a modularized frame using standardized parts reducing the number of single pieces by approximately 58% when compared to the company current cabin Lift construction.
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An Experiment on the Suitability of RAM for Test Case DesignWu, Hong January 2009 (has links)
To perform software testing at the early stages of software development process can save the cost and effort on finding and fixing defects. As the first stage of software development process, requirements engineering has been moved away from project-initiated requirements engineering towards requirements-initiated development in the last decade. This leads new challenges that it demands support for handling the requirements continually come in from multiple stakeholders on multiple abstraction levels instead of some specific customers. Requirements Abstraction Model was developed as a hierarchical abstraction method for requirements management, which is enable product management to leverage their resources and select requirements for implementation without overloading the organization. RAM was validated in industry on the usability for requirements management, but there is no evaluation for RAM on software testing. This thesis presents an empirical study with a goal of evaluating the suitability of RAM for test case design in respective of efficiency and effectiveness by the comparison with IEEE Std. 830 which is a standard of the traditional requirements specification. For achieving the goal of this study, a controlled experiment is conducted based on the refinement on an initial experiment planning, and is operated with twenty developers in industry in China. Analysis of the collected data from the experiment indicates that RAM has a similar effectiveness as using the requirements in IEEE Std. 830 format, while RAM is more efficient for test case design. Therefore, RAM is suitable for test case design, and has better performance than IEEE Std. 830 comprehensively in view of both efficiency and effectiveness.
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Textram, un lenguaje de especificación de dominio textual, para el desarrollo y manejo de modelos RAMOliva Rodríguez, Marel Josué January 2015 (has links)
Magíster en Tecnologías de la Información / La programación orientada a aspectos (AOP) sirve para separar las preocupaciones transversales desde la perspectiva de implementación de un sistema de software. El modelamiento orientado a aspectos (AOM) ofrece una alternativa para representar las preocupaciones transversales en niveles más altos de abstracción, en etapas anteriores a la implementación del software. "Reusable Aspect Models" (RAM) es un enfoque de AOM, que permite el diseño detallado de un sistema de software a través de módulos llamados "aspect models". La representación gráfica de RAM está inspirada por UML; los aspectos son definidos estáticamente a través de diagramas de clases y dinámicamente por medio de diagramas de estado y de secuencia. La consistencia entre modelos está asegurada por RAM, gracias a la
jerarquía de modelos y estructuras de dependencias. TouchRAM es una herramienta gráfica para el modelado de aspectos RAM. Actualmente TouchRAM continúa en desarrollo y es la única herramienta disponible para la creación de RAMs. TouchRAM no está disponible para el público general, dificultando su adopción en otros proyectos de software.
El objetivo de la presente tesis es presentar TextRAM, un modelador textual para la definición de modelos RAM. TextRAM se presenta como una alternativa a TouchRAM. El código fuente de TextRAM está disponible para su descarga y su implementación pretende estimular el desarrollo e investigación de RAM. Los modelos resultantes de TextRAM, podrán ser utilizados por TouchRAM y viceversa. Los usuarios de RAM, podrán elegir entre un modelamiento gráfico con TouhRAM o de un modelamiento textual con TextRAM.
Los objetivos de TextRAM son: (1) definición de una abstracción adecuada para la sintaxis concreta textual, (2) implementar una extensión de Eclipse que permita la edición de modelos con la sintaxis concreta de TextRAM, (3) tranformación de modelos desde TextRAM a TouchRAM y viceversa, (4) Aplicar las validaciones semánticas alineadas a las reglas de TouchRAM, (5) demostrar la validez de TextRAM, por medio de un caso de estudio llamado Slot Machines.
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