The zinc refinery for its non-conforming products in quality presents losses in the daily production that must be delivered to its main clients, with monetary losses that are quantified in $ 3,000,000 per year and the prestige before its client portfolio. This occurs due to a lack of controls, operational standards and discipline in the execution of the tasks of the operators in the production lines that guarantee the quality of zinc with a purity of 99.995%. The objective of the work carried out was to evaluate the economic losses and the reasons that these losses cause for the companies and an improvement action plan was designed to solve these problems under the implementation of Lean Manufacturing tools such as 5 S, TPM and standardization of procedures. The development of the tools included analyzing the current state, making a diagnosis and an action plan for each system with those responsible for each area, and finally measuring the effectiveness of the tools with a reduction in losses of 15% in the first year and so on. / Las refinerías de zinc presentan perdidas en la producción diaria debido a productos no conformes por su calidad, con pérdidas monetarias que se cuantifican en $3.000.000 por año y el prestigio ante su cartera de clientes. Esto ocurre por falta de controles, estándares operacionales y disciplina en la ejecución de las tareas de los operadores en las líneas de producción que garanticen la calidad del zinc con una pureza del 99,995 %. El objetivo del trabajo realizado fue evaluar las pérdidas económicas y los motivos que ocasionan las mismas. Se diseñó un plan de acción de mejora para solucionar estos problemas bajo la implementación de las herramientas de Lean Manufacturing como 5S, TPM y estandarización de procedimientos. El desarrollo de las herramientas abarco analizar el estado actual, realizar un diagnóstico y un plan de acción por cada sistema con los responsables de cada área y finalmente medir la efectividad de las herramientas con una previsión de reducción de las pérdidas del 15% en el primer año y así consecutivamente durante los cinco primeros años. / Trabajo de Suficiencia Profesional

SME companies are of greatest importance to our society. They contribute to job opportunities for a large portion of the population and account for 60 percent of the total revenue in the Swedish Enterprise. Given that there are a lot of SME:s, it is crucial to obtain efficient production.  In today's manufacturing industry, a major issue is the fact that value-added time is low compared to the total time it takes to manufacture a product. Lean production is a well-designed and proven method to apply to streamline production. With the help of Lean tools, continuous improvements can be made by eliminating unnecessary and inefficient processes in production. The aim of this study has been to investigate how SME:s can streamline their production using value stream analysis, and the questions that have been answered are:  - What methods can be used to evaluate production efficiency? - What parameters affect production efficiency? - How can a value stream analysis be used to streamline production and identify improvement measures?  The empirical case study was conducted using data collection methods that included interviews with employees at the case company, time measurements, and observations. These methods formed the basis for the creation of the present value analysis as well as the calculations of the OEE-values.  The result shows that SMEs can use value stream analysis as a way to identify inefficiencies and thereby implement improvement measures that increase production efficiency.  Keywords: SME, Lean, Value-stream mapping, Muda, 5S, RBV / Små- och medelstora företag är av yttersta vikt för vårt samhälle. Det bidrar till jobb åt en stor del av befolkningen och det bidrar till 60 procent av den totala omsättningen i det svenska näringslivet. Med hänsyn till att det finns så många SMF är det av yttersta vikt att erhålla en effektiv produktion. I dagens tillverkningsindustri är ett stort problem det faktum att den värdeskapande tiden är låg i jämförelse med den totala tiden det tar för att tillverka en produkt. Lean production är en välutformad och beprövad metod att tillämpa för att effektivisera en produktion. Med hjälp av Lean-verktyg kan ständiga förbättringar ske genom att eliminera onödiga och ineffektiva moment i produktionen. I studien har syftet varit att studera hur SMF kan effektivisera sin produktion med hjälp av en värdeflödesanalys och frågeställningarna som besvarats har varit: - Vilka metoder kan användas för att utvärdera produktionseffektiviteten? - Vilka parametrar påverkar produktionseffektiviteten? - Hur kan en värdeflödesanalys användas för att effektivisera produktionen och identifiera förbättringsåtgärder? Den empiriska fallstudien har genomförts med hjälp av datainsamling i form av intervjuer med anställda på fallföretaget, tidmätningar och observationer. De här metoderna har legat till grund för skapandet av nuvärdesanalysen samt beräkningar av TAK-värdena. Resultatet visar att SMF kan använda värdeflödesanalys som ett sätt för att identifiera ineffektivitet och därigenom genomföra förbättringsåtgärder som ökar produktionseffektiviteten.

The following study is a bachelor thesis at Jönköping Institute of Technology in collaboration with the lighting company Westal AB. The company had recently invested in a laser cutter and at the same time acquired the business Ledolight Sweden AB, which manufactures emergency exit luminaires. The problem that formed the basis of the study was that the responsible parties were unsure of how the machines should be positioned regarding proximity and a good working environment. The aimwas therefore to develop two concrete layout proposals for the workshop. Where one layout proposal should provide suggestions for the location of the laser cutter in relation to existing machines. While the layout proposal will provide a proposal on how the machines from Ledolight Sweden AB can be placed on a separate area in the workshop.The study's questions were answered using both quantitative and qualitative methods to obtain information. Value stream analyses, measurements, observations, and interviews are some of the methods used. Based on the chosen methods, it has been possible to describe a current situation at Westal AB and Ledolight Sweden AB. When designing the layout proposals, the collected information was compared to the selected theoretical framework. Subsequently, the "Simplified Systematic Layout Planning" (FSLP) method was used to identify and determine relationships, causes of proximity, space requirements and other needs.The study results in different layout proposals, which show how the machines and functions can be placed in the workshop. A total of three layout proposals have been produced for the laser cutter and two for the machines from Ledolight Sweden AB. Followed by the advantages and disadvantages of the chosen locations. In comparison with the current layout, the new layout proposals have achieved the purpose of creating proximity, creating good working conditions, and placing the machines from Ledolight Sweden AB in a separate area in the workshop. The study also provides a concrete proposal on how two of the layout proposals could be integrated into the workshop. The layout proposal contributes to less material handling as the times for moving materials have been reduced. Material handling has been reduced by 20% for the Stall product group. Regarding the product groups Gustav and LedoNomic material handling has been reduced by 13% and 47% respectively. In connection with the results of the study, the following recommendations have been identified,- Design spaghetti diagrams and eliminate unnecessary movements for all product groups processed in the workshop. - Apply the FSLP method to future layout changes.- Continued work with the Lean mindset and the tool 5S to eliminate waste. / Följande studie är ett examensarbete på Jönköpings tekniska högskola som utförts i samarbete med belysningsföretaget Westal AB. Företaget hade nyligen investerat i en laserskärare och samtidigt köpt upp verksamheten Ledolight Sweden AB som tillverkar nödutgångsarmaturer. Problemet som låg till grund för studien var att det inte fanns någon tydlig bild över hur maskinerna ska placeras med avseende på att skapa närhet och en god arbetsmiljö. Syftet var därmed att ta fram två konkreta layoutförslag över verkstaden. Där ena layoutförslaget ska ge förslag på placeringen av laserskäraren i förhållande till befintliga maskiner. Det andra layoutförslaget skall ge ett förslag på hur maskinerna från Ledolight Sweden AB kan placeras på en separat yta i verkstaden.Studiens frågeställningar har besvarats genom användandet av både kvantitativa och kvalitativa metoder för att inhämta information. Värdeflödesanalyser, mätningar och intervjuer är några av de metoderna som använts. Vid utformandet av layoutförslagen ställdes den inhämtade informationen i relation till det utvalda teoretiska ramverket.Därefter användes metoden ”Förenklad systematisk lokalplanläggning” (FSLP) för att ta fram olika de olika layoutförslagen.Studien resulterar i olika layoutförslag, vilket visar hur maskinerna respektive funktionerna kan placeras i verkstaden. Totalt har tre layoutförslag för laserskäraren tagits fram och två för maskinerna från Ledolight Sweden AB. Följt av fördelar respektive nackdelar med de valda placeringarna. I jämförelse med den nuvarande layouten, har de nya layoutförslagen uppnått syftet med att skapa närhet, skapa goda arbetsförhållanden och placerat maskinerna från Ledolight Sweden AB på en separat yta i verkstaden. Studien ger även ett konkret förslag på hur två av layoutförslagen skulle kunna integreras i verkstaden. Layoutförslaget bidrar till mindre materialhantering då tiderna för förflyttning av material har reducerats. Materialhanteringen har reducerats med 20% för produktgruppen Stall. Gällande produktgrupperna Gustav och LedoNomic materialhantering har det reducerats med 13% respektive 47%. I samband med studiens resultat har följande rekommendationer identifierats.- Utforma spaghettidiagram och eliminera onödiga rörelser för alla produktgrupper som förädlas i verkstaden. - Tillämpa metoden FSLP vid framtida layoutförändringar.- Fortsatt arbete med tankesättet Lean och verktyget 5S för att eliminera slöseriet.

This study is a research around an issue encountered by MyEasyLab, a laboratory involved in asbestos testing for the French market. They are facing a problem of sample losses, which is hardly acceptable, especially when growing on a highly competitive and price driven market. They are therefore looking for ways to have a better traceability of their samples. The technicians were observed to identify their needs and the problems the company is facing. This study is presenting as a result two complementary solutions: the reorganization of the working space, and the usage of the RFID (Radio Frequency Identification) technology to track the samples. The reorganization of the room using the 5S method should enable a more optimized way of working. This method initially used in the industry early on by Toyota, enables to optimize the flows (goods, person, waste …) in a sustainable way. The use of RFID presents the opportunity of a deep change in the whole process of the company, from the client to the labs. Bags already tagged RFID could be sent to the clients for their sample collection, enabling traceability from the beginning of the chain. The samples could, using RFID, be detected as they enter the site, leave the site and if they are in an undesired location (for example in the trash). The RFID technology is also an opportunity to facilitate the work of the technician, with a possibility to save around 4h15 of working time per day. The kind of tags that seems the most appropriate for this application is passive UHF tags. They present the advantages of being cheaper, easier to buy in bulk and readable from longer distances.  Even though the RFID technology is often seen as expensive, a return on investment seems possible for a tag price under 0.08€.  Further experimental tests need to be realized to refine the results of this study.

I en global ekonomi med ökade krav på konkurrenskraft och effektivitet är implementeringen av Lean-verktyg, såsom 5S, avgörande för att förbättra verksamhetens operationella prestanda. Trots detta uppstår ofta utmaningar vid implementering som kan påverka metodens framgång och effektivitet, särskilt när mänskliga aspekter försummas. Studien adresserar detta forskningsgap genom att undersöka engagemangets betydelse vid implementering av 5S inom processindustrin. Syftet med studien var att kartlägga hur mänskliga aspekter påverkar implementeringen av 5S inom processindustrin samt ge förslag på förbättringsåtgärder för att stärka engagemanget vid förändringsprocesser. Studien använde ett vetenskapligt angreppssätt med en tydlig problemformulering och ett klart definierat syfte. En fallstudie av ett företag, vilka är aktiva inom processindustrin, valdes på grund av dess pågående implementering av 5S. Data samlades in både kvalitativt och kvantitativt genom semistrukturerade intervjuer med ledningen för 5S-arbetet och enkätundersökningar med operatörerna. Reliabiliteten säkerställdes genom standardisering av datainsamlingsmetoderna, medan validiteten förstärktes genom datatriangulering och forskartriangulering. Etiska aspekter hanterades enligt Vetenskapsrådets riktlinjer, vilket inkluderade informationskrav, samtyckeskrav, konfidentialitetskrav och nyttjandekrav. Fallstudieprotokoll och systematisk datainsamling användes för att stärka studiens trovärdighet och generaliserbarhet. Studiens resultat visar att stöd från både ledning och organisation är avgörande för att öka engagemanget. Operatörernas upplevelser av rättvisa, återkoppling och belöningssystem identifieras som centrala drivkrafter för att främja ett hållbart engagemang. Många operatörer och tjänstemän kände sig inte involverade i beslutsprocesserna, vilket skapade motstånd mot 5S-initiativet. Informella ledare identifierades som avgörande för att påverka teamens inställning och engagemang. Studien betonar även vikten av tydlig kommunikation, ledarskap och delaktighet i förändringsprocesser för en framgångsrik implementering av 5S. Slutsatsen av studien betonar att en framgångsrik implementering av 5S kräver ett holistiskt synsätt där både tekniska och mänskliga aspekter beaktas. Identifierade nyckelfaktorer för att stärka engagemanget inkluderar organisationens- och ledningens stöd, konstruktiv återkoppling, motivering och delaktighet samt säkerställande av tydlig och konsekvent kommunikation. Genom att adressera dessa aspekter kan företag i processindustrin, inte bara förbättra sin operationella prestanda, utan även öka medarbetarnöjdheten och nå framgång vid förändringsarbeten. / In a global economy with increasing demands for competitiveness and efficiency, the implementation of Lean tools, such as 5S, is crucial for enhancing operational performance. Despite this, challenges often arise during implementation, which can affect the method's success and efficiency, particularly when human aspects are neglected. This study addresses this research gap by examining the significance of engagement during the implementation of 5S in the process industry. The purpose of the study was to map how human aspects influence the implementation of 5S in the process industry and to propose improvement measures to strengthen engagement during change processes. The study employed a scientific approach with a clear problem formulation and a well-defined purpose. A case study of a company, which is active in the process industry, was selected due to its ongoing implementation of 5S. Data were collected both qualitatively and quantitatively through semi-structured interviews with the management of the 5S initiative and surveys with operators. Reliability was ensured through the standardization of data collection methods, while validity was enhanced through data triangulation and researcher triangulation. Ethical aspects were handled according to the guidelines of the Swedish Research Council, including requirements for information, consent, confidentiality, and usage. Case study protocols and systematic data collection were used to strengthen the study's credibility and generalizability. The study's results show that support from both management and the organization is crucial for increasing engagement. Operators' perceptions of fairness, feedback, and reward systems are identified as central drivers for promoting sustainable engagement. Many operators and officials felt uninvolved in decision-making processes, which created resistance to the 5S initiative. Informal leaders were identified as crucial for influencing teams' attitudes and engagement. The study also emphasizes the importance of clear communication, leadership, and participation in change processes for a successful implementation of 5S. The conclusion of the study emphasizes that a successful implementation of 5S requires a holistic approach that considers both technical and human aspects. Identified key factors for strengthening engagement include support from the organization and management, constructive feedback, motivation and participation, and ensuring clear and consistent communication. By addressing these aspects, companies in the process industry, not only improve their operational performance but also increase employee satisfaction and achieve success in change initiatives.

This is a study in how rooms with several activities within health and well-being can be designed using the 5S-methodology to facilitate the structure and order. The purpose of the study is to create structured and organized environment that highlights activities and functions. The study is a collaboration with Munktell Exercise & Health, a health- and fitness company in Eskilstuna with plans to develop the business with a new mindfulnessarea. Information has been gathered by methods like qualitative interviews, site analysis and analysis of referenceobjects. The premises along with previous examples of 5S-methodology and the requirements and wishes of mindfulness practice showed that there is a need for structure and organization in environments with multiple tasks and users. Theories about cognitive theory, visual balance, immerssion, affordance, signifiers and 5S-methodology shown examples of how people are affected by their environment. For example, the experience of a room and interpretation of the functions can be controlled in terms of organization and spatial attributes. In the current situation there are empty rooms available for the new part to take place and the realization of it. The result of the study is a designproposal from a human-centered design approach, combined with the 5S-methodology, where the 5S-methodology may be of interest to other training activities or organizations in the health sector in need of structure and organization. The designproposal is designed on the basis of the collected empirical data with the aim of preventing confusion and facilitate the users on the site with user-customized solutions and clarity of the rooms features. / Det här arbetet är en studie i hur rum med flera aktiviter inom hälsa och välmående kan utformas med hjälp av 5S-metoden för att underlätta för struktur och ordning. Syftet med studien är att skapa strukturerad och organiserad miljö som tydliggör aktiviteter och funktioner. Arbetet sker i samarbete med Munktell Träning & Hälsa, ett friskvårdsföretag i Eskilstuna som planerar att utveckla sin verksamhet med en ny mindfulnessdel. Med metoder som kvalitativa intervjuer, platsanalys och analys av referensobjekt har informations samlats kring de aktuella lokalerna, tidigare exempel av 5S-metoden samt krav och önskemål inom mindfulnessutövning som visat att det finns ett behov av struktur och organisering i miljöer med flera aktiviteter och användare. Utifrån gestalt- och schemateorier, visuell balans, immerssion, affordance och signifiers samt 5S-metoden visas exempel på hur människan påverkas av sin omgivning. Exempelvis hur upplevelsen av ett rum och tolkningen av funktioner kan styras i form av organisering och rumsliga attribut. I dagsläget finns tomma lokaler till förfogande för den nya delen och det planeras en ombyggnation i samband med förverkligandet av det. Arbetet mynnar ut i ett gestaltningsförslag från ett human centered design-perspektiv i kombination med 5S-metoden, där främst 5S-metoden kan vara av intresse för andra träningsverksamheter eller organisationer inom hälsosektorn som är i behov av strukturering och organisering. Gestaltningsförslaget är formgivet utifrån den insamlade empirin med syftet att förebygga oreda och underlätta för användarna på platsen med användaranpassade lösningar och tydlighet av rummens funktioner.

El sector textil y confecciones es uno de los sectores con mayores expectativas de crecimiento sostenible de la industria y uno de los que mayor competencia acumula con otros países del sector internacional. Este sector abarca diferentes tipos de procesos, que van desde tratamiento de fibras, hilatura, tejido y confección. El presente análisis recae sobre la baja disponibilidad que presenta los equipos en el proceso de teñido. Este proceso se mide a partir de los reprocesos que existen debido a que se ejecuta mantenimientos correctivos, el cual afecta la producción en el teñido de telas. Asimismo, se logra evaluar el tiempo promedio entre fallas (MTBF) y el tiempo promedio de reparación (MTTR), lo que indica la variabilidad por debajo del objetivo esperado por la organización. Por otro lado, en el tercer capítulo se desarrolla le propuesta de mejora a partir del desarrollo del estado del arte del primer capítulo. Para ello se propone la metodología da seguir de acuerdo con una combinación de RCM y TPM. Bajo esta metodología se desarrolla diferentes actividades y estrategias que permitirán detectar equipos críticos e implementar planes de mantenimiento preventivo mecánico y eléctrico. Además, se incluye como parte de este sistema la implementación de un mantenimiento basado en condición (predictivo) y autónomo. Finalmente, el tercer capítulo finaliza con el soporte de la metodología 5 ‘’S’’ y la evaluación económica a partir de la estructura de costos que tiene prevista el proyecto. Finalmente, en el capítulo cuatro se presenta la validación de la propuesta de mejora a partir de la validación de los resultados en el software Arena. Ello permite concluir que los indicadores MTTR y MTBF ha logrado alcanzar los objetivos presentados en la propuesta. / The textile and garment sector is one of the sectors with the highest expectations of sustainable growth in the industry and one of the sectors with the highest competition with other countries in the international sector. This sector covers different types of processes, ranging from fiber treatment, spinning, weaving and confection. The present analysis falls on the low availability that presents the equipment in the process of dyeing. This process is measured from the reprocesses that exist due to the execution of corrective maintenance, which affects the production of fabric dyeing. Likewise, it is possible to evaluate the average time between failures (MTBF) and the average repair time (MTTR), which indicates the variability below the target expected by the organization. On the other hand, in the third chapter the improvement proposal is developed from the development of the state of art of the first chapter. For this, the methodology to be followed is proposed according to a combination of RCM and TPM. Under this methodology, different activities and strategies are developed to detect critical equipment and implement mechanical and electrical preventive maintenance plans. In addition, the implementation of a maintenance based on (predictive) and autonomous condition is included as part of this system. Finally, the third chapter ends with the support of the 5 '' S '' methodology and the economic evaluation based on the cost structure planned by the project. Finally, chapter four presents the validation of the improvement proposal based on the validation of the results in the Arena software. This allows concluding that the MTTR and MTBF indicators have achieved the objectives presented in the proposal. / Tesis

Submitted by Angela Maria de Oliveira (amolivei@uepg.br) on 2017-10-31T12:38:31Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇÃO michele.pdf: 3067898 bytes, checksum: 4e684091426aed5eb28e585d5db27abb (MD5) / Made available in DSpace on 2017-10-31T12:38:31Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇÃO michele.pdf: 3067898 bytes, checksum: 4e684091426aed5eb28e585d5db27abb (MD5) Previous issue date: 2014-02-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Alticinae apresenta características cariotípicas muito interessantes quanto a variação do número diplóide, do sistema de determinação sexual e irregularidades meióticas. O número cromossômico mais frequente é de 11 ou 12 pares. As espécies de Oedionychina estudadas citogeneticamente possuem 2n= 22,10II+X+Y com cromossomos sexuais gigantes. No que se refere à posição sistemática existem muitas divergências entre os estudos e problemas de identificação das espécies pertencentes aos vários gêneros. O estudo com técnicas mais refinadas e o mapeamento do DNA r 5S é recente em Coleoptera, e as poucas espécies de Alticinae estudadas mostram a presença de dois ou três pares autossômicos. Assim, este trabalho tem o objetivo de analisar citogeneticamente e propor as estratégias de diferenciação cariotípica para as espécies de Omophoita communis e Omophoita sexnotata. A análise citogenética dos indivíduos de duas populações de O. communis estudadas mostrou a ocorrência de grande variação no número diploide e morfologia cromossômica, sendo possível separá-las em dois citótipos. O citótipo I possui 2n= 22 e o citótipo II 2n= 12, sendo essa variação é descrita pela primeira vez no gênero. Adicionalmente, o estudo da morfologia do edeago mostrou diferenças, indicando um provável padrão de diferenciação das duas espécies. A análise da árvore consenso da reconstrução filogenética também evidencia que os citótipos mostram agrupamentos diferentes e reforçam a hipótese de duas espécies. O estudo do mapeamento do gene DNAr 5S em O. sexnotata evidenciou a presença desse cluster em todos os cromossomos autossômicos, sendo esse padrão de dispersão nunca descrito em Coleoptera. Na literatura, a dispersão dos genes ribossomais está sempre relacionada com a presença de elementos transponíveis. O resultado do sequenciamento dos fragmentos de 5S ribossomal obtidos de cada cromossomo de O. sexnotata resultaram em sequencias similares a RNAr 5S de Drosophila melanogaster, elemento transponível EnSpm, retropseudogene de 5S e microssatélite. Adicionalmente, a analise da estrutura secundaria do RNAr 5S, mostrou que as sequencias obtidas não são funcionais quando comparadas seus percentuais de energia livre em relação ao percentual da sequencia original do RNAr 5S. / Alticinae presents karyotypic characteristics very interesting as the variation of the diploid number, sex determination system and meiotic irregularities. The most frequent chromosome number is 11 or 12 pairs. The species cytogenetically studied Oedionychina have 2n = 22,10 II + X + Y with giant sex chromosomes. In relation to the systematic position there are many divergence between the studies and problems of identification of species belonging to several genera. The study with more refined techniques and rDNA 5s mapping is recent in Coleoptera , and the few Alticinae species studied show the presence of two or three autosomal pairs. Thus, this study aims to analyze cytogenetically and propose strategies for the species karyotype differentiation of Omophoita communis and Omophoita sexnotata. Cytogenetic analysis of individuals of both populations of O. communis showed the existence of a large variation in diploid number and chromosome morphology, being possible to separate them into two cytotypes. Cytotype I presented 2n = 22 and cytotype II 2n = 12, this variation is described for the first time in the genus. Additionally, the study of the morphology of the aedeagus showed differences, indicating a likely pattern of differentiation of the two species. The phylogenetic reconstruction consensus tree analysis also presented that cytotypes show different groupings and reinforce the hypothesis of two species. The 5S rDNA gene mapping of O. sexnotata showed the presence of this cluster in all autosomes, this dispersal pattern was never described in Coleoptera before. In the literature, the dispersion of ribosomal genes is always associated with the presence of transposable elements. The sequencing of 5S rRNA fragments obtained from each chromosome of O. sexnotata resulted in similar to 5S rRNA sequences of Drosophila melanogaster transposable element EnSpm, retropseudogene 5S and microsatellite. Additionally, analysis of the secondary structure of 5S rRNA showed that the sequences obtained are not functional compared their percentage of free energy with the percentage of the original sequence of the 5S rRNA .

Η ΒΥΠ διαθέτει αντίτυπο της διατριβής σε έντυπη μορφή στο βιβλιοστάσιο διδακτορικών διατριβών που βρίσκεται στο ισόγειο του κτιρίου της. / Στα βακτήρια, η μεγάλη ριβοσωματική υπομονάδα αποτελείται από δύο είδη RNA, το 23S και το 5S rRNA, καθώς και 33 πρωτεΐνες. Ο σχηματισμός του πεπτιδικού δεσμού και η απελευθέρωση της πεπτιδικής αλυσίδας επιτελούνται στη μεγάλη υπομονάδα, όπου εδράζεται το καταλυτικό κέντρο της πεπτιδυλοτρανσφεράσης (PTase). Εκτός αυτού, η μεγάλη υπομονάδα περιλαμβάνει το κέντρο προσδέσεως των μεταφραστικών παραγόντων, το οποίο πυροδοτεί τη GTPase δραστηριότητα των G-πρωτεϊνικών παραγόντων, που εμπλέκονται στη μετατόπιση των υποστρωμάτων και άλλες ριβοσωματικές λειτουργίες. Έχει υποτεθεί ότι το 5S rRNA παίζει ουσιώδη ρόλο στη συγκρότηση του κέντρου της PTase και στη μετάδοση σημάτων μεταξύ του καταλυτικού κέντρου και των ριβοσωματικών συστατικών που διεκπεραιώνουν τη μετατόπιση των υποστρωμάτων. Το ιοντικό περιβάλλον φαίνεται να επηρεάζει καθοριστικά τη διαμόρφωση του 5S rRNA. Για παράδειγμα, έχει βρεθεί ότι οι πολυαμίνες δεσμεύονται εκλεκτικά στο 5S rRNA και επηρεάζουν τη δραστικότητα του έναντι του διμεθυλο-θεϊκού, ενός αντιδραστηρίου-ιχνηθέτη της τριτοταγούς δομής του RNA. Αρχικά ελέγξαμε αν υπάρχουν εξειδικευμένες θέσεις πρόσδεσης των πολυαμινών στο 5S rRNA. Στη συνέχεια, με σκοπό να ελέγξουμε αν η πρόσδεση των πολυαμινών επηρεάζει τη λειτουργία του 5S rRNA, 70S ριβοσώματα προγραμματισμένα με πολύ-ουριδυλικό σχηματίσθηκαν από 50S υπομονάδες, ολικά ή εκλεκτικά φωτοσημασμένες με ένα φωτοδραστικό ανάλογο της σπερμίνης και από 30S ακατέργαστες υπομονάδες. Αυτά τα ριβοσώματα είχαν την ικανότητα να δεσμεύουν AcPhe-tRNA ελαφρώς ισχυρότερα, σε σύγκριση με ριβοσώματα συγκροτημένα από φυσικά συστατικά, μη περιέχοντα πολυαμίνες. Το γεγονός αυτό υποδηλώνει ότι η πρόσδεση πολυαμινών στο 5S rRNA επηρεάζει, σε μικρό βαθμό, τη λειτουργία του παράγοντα επιμήκυνσης EF-Tu. Συζευγμένα, όμως, τα εν λόγω ριβοσώματα με tRNAPhe στην Ε-θέση και AcPhe-tRNAστην Ρ-θέση, επέδειξαν ισχυρότερη καταλυτική δραστικότητα και αυξημένη ικανότητα για μετατόπιση των υποστρωμάτων. Τα αποτελέσματα αυτά εισηγούνται σημαντική εμπλοκή των πολυαμινών στο λειτουργικό ρόλο του 5S rRNA κατά την κατάλυση και μετατόπιση των υποστρωμάτων. / In bacteria, the large ribosomal subunit comprises two RNA species, 23S and 5S rRNA, and 33 proteins. Peptide bond formation and peptide release are catalyzed by the large subunit, where the peptidyltransferase (PTase) center is located. In addition to this center, which triggers the GTPase activities of G-protein factors involved in translocation and other ribosomal functions. It has been hypothesized that 5S rRNA plays essential role in assembling the PTase center and mediating signal transmissions between this center and the translocation machinery. Furthermore, the ionic environment seems to affect the conformation of 5S rRNA. For instance, polyamines have been found to bind specifically to 5S rRNA and influence the 5S rRNA reactivity towards dimethyl-sulfate (DMS), a chemical probe of RNA tertiary structure. Initially we examined whether there are specific sites for binding of polyamines. To test whether the binding of polyamines influence the function of 5S rRNA poly(U)-programmed 70S ribosomes were reconstituted from 50S subunits, totally or specifically photolabelled in their 5S rRNA with a photoreactive analogue of spermine, and native 30S subunits. These ribosomes were found to enzymatically bind AcPhe-tRNA better than ribosomes reconstituted from native components. This means, that furnishing 5S rRNA with spermine slightly influences the elongation factor EF-Tu function. However, equipped with tRNAPhe at the A-site and AcPhe-tRNA at the P-site, these ribosomes exhibited higher catalytic activity and enhanced tRNA translocation efficiency. These results suggest an essential impact of polyamines on the functional role of 5S rRNA in catalysis and translocation of translation substrates.

Orientador: Adriane Pinto Wasko / Resumo: O Brasil apresenta umas das maiores diversidades de avifauna do mundo, sendo estimada a ocorrência de 1.919 espécies distribuídas em todo seu território. Entretanto, a destruição de habitats, poluição e captura excessiva, muitas vezes associadas ao comércio ilegal, têm levado a um declínio no número de indivíduos desse grupo animal. Uma das formas de combater esse tipo de crime ambiental e de reverter ou minimizar seus efeitos concerne à implementação de um banco de dados forenses com registros de ocorrências e informações biológicas sobre os animais apreendidos e desenvolvimento de ações de conscientização da população sobre esta problemática. Desta forma, o presente trabalho teve como objetivo desenvolver estratégias multidisciplinares para a proteção jurídica à conservação biológica de aves oriundas do tráfico de animais. Amostras de DNA de aves comercializadas ilegalmente e encaminhadas ao Centro de Medicina e Pesquisa em Animais Silvestres (CEMPAS), mantido pela Faculdade de Medicina Veterinária e Zootecnia da Universidade Estadual Paulista, foram utilizadas para geração de perfis genéticos sexo-específicos, por meio da amplificação de segmentos dos genes CHD-Z e CHD-W (Chromo Helicase DNA Binding). Adicionalmente, perfis de duas subespéciesde papagaio-verdadeiro (Amazona aestiva aestiva e Amazona aestiva xanthopteryx foram gerados por meio da amplificação e sequenciamento nucleotídico de segmentos dos genes mitocondriais citocromo C oxidase subunidade I (COI) e citocrom... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Brazil has one of the largest avifauna diversity in the world, with estimated 1,919 species distributed throughout its territory. However, habitat destruction, pollution and over-harvesting, often associated with illegal trade, have led to a decline in the number of individuals of this animal group. One possible approach to combat this type of environmental crime and to reverse or minimize its effects concerns the implementation of a forensic database with records of occurrences and biological information about the captured animals and the development of actions to raise population awareness about this problem. Therefore, the present work aimed to develop multidisciplinary strategies for the legal protection to the biological conservation of birds originated from the animal traffic. DNA samples of illegally traded birds, that were sent to the Center for Medicine and Research in Wild Animals (CEMPAS) maintained by the Faculty of Veterinary Medicine and Animal Science of the São Paulo State University, were used to generate sex-specific genetic profiles, through the amplification of segments of the CHD-Z and CHD-W (Chromo Helicase DNA Binding) genes. In addition, profiles of two subspecies of the Blue-fronted Amazon (Amazona aestiva aestiva and Amazona aestiva xanthopteryx) were generated, through amplification and nucleotide sequencing of segments of the mitochondrial genes cytochrome C oxidase subunit I (COI) and cytochrome b (CYB) and a segment of the 5S ribosomal DNA gene (... (Complete abstract click electronic access below) / Doutor