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The dissertation analyses the reliefs and paintings on thirty-one different tombs in Western Anatolia erected between 600 and 450 BC, in order to illuminate the ways in which non-Greek elites were identified on their memorials. The tombs from three areas are treated: Phrygia, Hellespontine Phrygia and Lydia, where the primary language groups were Phrygian, Mysian and Lydian. There is little literary evidence for these regions, and what there is tends to focus on political developments. Descriptions of people and society are few, and tend to represent them from an outside perspective, grouping them according to cultural characteristics which differentiate them from Greeks. It is clear, however, that the regions were important, prosperous places, controlled by illustrious grandees and land marked with a relatively high proportion of monumental tombs. Of these monumental tombs, there is a relatively high number decorated with striking and articulate images. There is much to be gained from examining the images on these tombs, as ‘indigenous’ sources for how elite Western Anatolians described themselves. Previous approaches to the tombs and their images have tended to look at them individually or in smaller groups, and to concentrate on the transmission and reception of Persian and Greek culture in the Achaemenid provinces. This dissertation contributes a broader comparative study of the decorated tombs, focussing on the kinds of statuses the images represent and the cultural forms these took. By comparing the various methods of self-representation, it clarifies patterns of identities in Western Anatolia and their relationship to historical circumstances. The dissertation is divided into five chapters. An introduction outlines the scope and sample, the historical background, previous studies of the monuments, the definition of ‘identity’ and the methods of analysis adopted here. Three case study chapters present the regions and the decorated monuments within them. A concluding chapter synthesises three aspects: social identities (roles and spheres of life represented); geographic and chronological patterns; and cultural affiliations and orientations. The dissertation concludes that a tension between Persian identities and local traditions is evident in some of the tomb images, which relates to the political upheavals in Western Anatolia and the Aegean at the time of the Persian Wars.

Em procariotos, RNAs antisense (asRNAs) constituem a classe de RNAs não codificantes (ncRNAs) mais numerosa detectada por métodos de avaliação de transcritoma em larga escala. Apesar da grande abundância, pouco se sabe sobre mecanismos regulatórios e aspectos da conservação evolutiva dessas moléculas, principalmente em arquéias, onde o mecanismo de degradação de RNAs dupla fita (dsRNAs) é um fenômeno pouco conhecido. No presente estudo, utilizando dados de dRNA-seq, identificamos 1626 inícios de transcrição primários antisense (aTSSs) no genoma de Halobacterium salinarum NRC-1, importante organismo modelo para estudos de regulação gênica no domínio Archaea. Integrando dados de expressão gênica obtidos a partir de 18 bibliotecas de RNA-seq paired-end, anotamos 846 asRNAs a partir dos aTSSs mapeados. Encontramos asRNAs em ~21% dos genes anotados, alguns desses relacionados a importantes características desse organismo como: codificadores de proteínas que constituem vesículas de gás e da proteína bacteriorodopsina, além de vários genes relacionados a maquinaria de tradução e transposases. Além desses, encontramos asRNAs em genes pertencentes a sistemas de toxinas-antitoxinas do tipo II e utilizando dados públicos de dRNA-seq, evidenciamos que esse é um fenômeno que ocorre em bactérias e arquéias. A interação de um ncRNA com seu RNA alvo pode ser dependente de proteínas, em arquéias, a proteína LSm é uma chaperona de RNA homóloga a Hfq de bactérias, implicada no controle pós-transcricional. Utilizamos dados de RIP-seq de RNAs imunoprecipitados com LSm e identificamos 91 asRNAs interagindo com essa proteína, para 81 desses, o mRNA do gene sense também foi encontrado interagindo. Buscando por aTSSs presentes nas mesmas regiões de genes ortólogos, identificamos 160 aTSSs que dão origem a asRNAs em H. salinarum possivelmente conservados em Haloferax volcanii. A expressão dos asRNAs anotados foi avaliada ao longo de uma curva de crescimento e em uma linhagem knockout de um gene que codifica uma RNase R, possível degradadora de dsRNAs em arquéias. Encontramos um total de 144 asRNAs diferencialmente expressos ao longo da curva de crescimento, para 56 desses o gene sense também está diferencialmente expresso, caracterizando possíveis mecanismos de regulação em cis por esses RNAs. Na linhagem knockout, encontramos cinco asRNAs diferencialmente expressos e apenas para um desses o gene sense também está diferencialmente expresso, resultado que não nos permitiu inferir um possível papel de degradação de dsRNAs da RNAse R em H. salinarum NRC-1. Nesse trabalho apresentamos um mapeamento completo do transcritoma antisense primário de H. salinarum NRC-1 com resultados que consistem em um importante passo na direção da compreensão do envolvimento da transcrição antisense na regulação gênica pós-transcricional desse organismo modelo do terceiro domínio da vida. / Antisense RNAs (asRNAs) constitute the most numerous class of non-coding RNAs (ncRNAs) detected by transcriptome highthroughput methods in prokaryotes. Despite this abundance, little is known about regulatory mechanisms and evolutionary aspects of these molecules, mainly in archaea, where the mechanism of double-strand RNA (dsRNA) degradation remains poorly understood. In this study, using dRNA-seq data, we identified 1626 antisense transcription start sites (aTSSs) in the genome of Halobacterium salinarum NRC-1, an important model organism for gene expression regulation studies in Archaea. By integrating gene expression data from 18 RNA-seq paired-end libraries, we were able to annotate 846 asRNAs from mapped aTSSs. We found asRNAs in ~21% of annotated genes including genes related to important characteristics of this organism, such as: gas vesicle proteins, bacteriorhodopsin, translation machinery and transposases. We also found asRNAs in type II toxin-antitoxin systems and using public dRNA-seq data, we show evidences that this phenomenon might be conserved in archaea and bacteria. The interaction of a ncRNA with its target may depend on intermediary proteins action. In archaea, the LSm protein is a RNA chaperone homologous to bacterial Hfq, involved in post-transcriptional regulation. We used RIP-seq data from RNAs immunoprecipitated with LSm and identified 91 asRNAs interacting with this protein, for 81 of these the mRNA of the sense gene is also interacting. We searched for aTSSs present in the same region of orthologous genes in the Haloferax volcanii. We found 160 aTSSs that originated asRNAs in H. salinarum NRC-1 that might be conserved in this two archaea. The expression of annotated asRNAs was analyzed over a growth curve and in a knockout strain for RNase R gene. We found 144 asRNA differentially expressed over the growth curve, for 56 of these the sense gene was also differentially expressed, characterizing possible cis regulators asRNAs. In the knockout strain we found five differentially expressed asRNAs and only one asRNA/gene pair, this result does not allow us to infer a dsRNA degradation in vivo activity for this RNase in H. salinarum NRC- 1. This work contributes to the discovery of the antisense transcriptome in H. salinarum NRC- 1 a relevant step to uncover the post-transcriptional gene regulatory network in this archaeon.

Em procariotos, RNAs antisense (asRNAs) constituem a classe de RNAs não codificantes (ncRNAs) mais numerosa detectada por métodos de avaliação de transcritoma em larga escala. Apesar da grande abundância, pouco se sabe sobre mecanismos regulatórios e aspectos da conservação evolutiva dessas moléculas, principalmente em arquéias, onde o mecanismo de degradação de RNAs dupla fita (dsRNAs) é um fenômeno pouco conhecido. No presente estudo, utilizando dados de dRNA-seq, identificamos 1626 inícios de transcrição primários antisense (aTSSs) no genoma de Halobacterium salinarum NRC-1, importante organismo modelo para estudos de regulação gênica no domínio Archaea. Integrando dados de expressão gênica obtidos a partir de 18 bibliotecas de RNA-seq paired-end, anotamos 846 asRNAs a partir dos aTSSs mapeados. Encontramos asRNAs em ~21% dos genes anotados, alguns desses relacionados a importantes características desse organismo como: codificadores de proteínas que constituem vesículas de gás e da proteína bacteriorodopsina, além de vários genes relacionados a maquinaria de tradução e transposases. Além desses, encontramos asRNAs em genes pertencentes a sistemas de toxinas-antitoxinas do tipo II e utilizando dados públicos de dRNA-seq, evidenciamos que esse é um fenômeno que ocorre em bactérias e arquéias. A interação de um ncRNA com seu RNA alvo pode ser dependente de proteínas, em arquéias, a proteína LSm é uma chaperona de RNA homóloga a Hfq de bactérias, implicada no controle pós-transcricional. Utilizamos dados de RIP-seq de RNAs imunoprecipitados com LSm e identificamos 91 asRNAs interagindo com essa proteína, para 81 desses, o mRNA do gene sense também foi encontrado interagindo. Buscando por aTSSs presentes nas mesmas regiões de genes ortólogos, identificamos 160 aTSSs que dão origem a asRNAs em H. salinarum possivelmente conservados em Haloferax volcanii. A expressão dos asRNAs anotados foi avaliada ao longo de uma curva de crescimento e em uma linhagem knockout de um gene que codifica uma RNase R, possível degradadora de dsRNAs em arquéias. Encontramos um total de 144 asRNAs diferencialmente expressos ao longo da curva de crescimento, para 56 desses o gene sense também está diferencialmente expresso, caracterizando possíveis mecanismos de regulação em cis por esses RNAs. Na linhagem knockout, encontramos cinco asRNAs diferencialmente expressos e apenas para um desses o gene sense também está diferencialmente expresso, resultado que não nos permitiu inferir um possível papel de degradação de dsRNAs da RNAse R em H. salinarum NRC-1. Nesse trabalho apresentamos um mapeamento completo do transcritoma antisense primário de H. salinarum NRC-1 com resultados que consistem em um importante passo na direção da compreensão do envolvimento da transcrição antisense na regulação gênica pós-transcricional desse organismo modelo do terceiro domínio da vida. / Antisense RNAs (asRNAs) constitute the most numerous class of non-coding RNAs (ncRNAs) detected by transcriptome highthroughput methods in prokaryotes. Despite this abundance, little is known about regulatory mechanisms and evolutionary aspects of these molecules, mainly in archaea, where the mechanism of double-strand RNA (dsRNA) degradation remains poorly understood. In this study, using dRNA-seq data, we identified 1626 antisense transcription start sites (aTSSs) in the genome of Halobacterium salinarum NRC-1, an important model organism for gene expression regulation studies in Archaea. By integrating gene expression data from 18 RNA-seq paired-end libraries, we were able to annotate 846 asRNAs from mapped aTSSs. We found asRNAs in ~21% of annotated genes including genes related to important characteristics of this organism, such as: gas vesicle proteins, bacteriorhodopsin, translation machinery and transposases. We also found asRNAs in type II toxin-antitoxin systems and using public dRNA-seq data, we show evidences that this phenomenon might be conserved in archaea and bacteria. The interaction of a ncRNA with its target may depend on intermediary proteins action. In archaea, the LSm protein is a RNA chaperone homologous to bacterial Hfq, involved in post-transcriptional regulation. We used RIP-seq data from RNAs immunoprecipitated with LSm and identified 91 asRNAs interacting with this protein, for 81 of these the mRNA of the sense gene is also interacting. We searched for aTSSs present in the same region of orthologous genes in the Haloferax volcanii. We found 160 aTSSs that originated asRNAs in H. salinarum NRC-1 that might be conserved in this two archaea. The expression of annotated asRNAs was analyzed over a growth curve and in a knockout strain for RNase R gene. We found 144 asRNA differentially expressed over the growth curve, for 56 of these the sense gene was also differentially expressed, characterizing possible cis regulators asRNAs. In the knockout strain we found five differentially expressed asRNAs and only one asRNA/gene pair, this result does not allow us to infer a dsRNA degradation in vivo activity for this RNase in H. salinarum NRC- 1. This work contributes to the discovery of the antisense transcriptome in H. salinarum NRC- 1 a relevant step to uncover the post-transcriptional gene regulatory network in this archaeon.

Le trouble d’anxiété de séparation (TAS) est le trouble anxieux le plus prévalent chez les enfants. Il apparaît tôt et entraîne plusieurs conséquences négatives. La thérapie cognitivo-comportementale (TCC) a été reconnue efficace pour traiter les troubles anxieux. Toutefois, peu d’études ont vérifié son efficacité pour le traitement spécifique du TAS et très peu en ont examiné l’effet auprès d’enfants de moins de 7 ans. Les quelques interventions étudiées visant les moins de 7 ans ont en commun d’inclure le parent dans le traitement ou de l’offrir directement à celui-ci. L’objectif principal de cette thèse est de vérifier l’efficacité d’un programme d’entraînement parental de type TCC, adapté pour les parents d’enfants de 4 à 7 ans souffrant de TAS. Cette étude vise également deux objectifs spécifiques : observer la fluctuation des symptômes de TAS de l’enfant pendant le traitement et examiner l’impact du programme sur les variables parentales. Un devis à cas unique à niveaux de base multiples a été utilisé. Six familles ont pris part à l’étude. Des entrevues semi-structurées, des questionnaires auto-administrés et des calepins d’auto-observations quotidiennes ont été utilisés auprès des parents pour mesurer les symptômes anxieux des enfants, leurs pratiques parentales, le stress parental et leurs symptômes anxieux et dépressifs. Des questionnaires sur les difficultés de l’enfant incluant l’anxiété ont aussi été envoyés à l’éducatrice ou à l’enseignante. Tous les questionnaires ont été administrés aux trois temps de mesure (prétraitement, post-traitement et relance 3 mois). Les calepins d’auto-observations ont été remplis quotidiennement durant le niveau de base, pour toute la durée de l’intervention et pendant une à deux semaines à la relance. Les résultats de l’étude indiquent que cinq enfants sur six ne répondent plus au diagnostic de TAS suite au traitement ainsi que trois mois plus tard. Les résultats des calepins d’auto-observations montrent une amélioration claire des manifestations principales de TAS pour la moitié des enfants et plus mitigée pour l’autre moitié, de même qu’une amélioration systématique de la fréquence hebdomadaire totale de manifestations de TAS suite à l’intervention pour quatre enfants. Les résultats aux questionnaires remplis par les parents montrent une amélioration des symptômes d’anxiété et de TAS chez quatre enfants au post-test et/ou à la relance, tandis que les questionnaires de l’éducatrice (ou enseignante) suggèrent que les symptômes anxieux des enfants se manifestaient peu dans leur milieu scolaire ou de garde. L’impact du programme sur le stress parental et les pratiques parentales est également mitigé. Ces résultats suggèrent que le Programme d’entraînement parental pour les enfants souffrant d’anxiété de séparation (PEP-AS) est efficace pour réduire les symptômes de TAS chez les enfants d’âge préscolaire ou en début de parcours scolaire et appuient la pertinence d’offrir le traitement aux parents et d’inclure un volet relationnel. D’autres études seront cependant nécessaires pour répliquer ces résultats auprès d’un plus vaste échantillon. Il serait également intéressant de vérifier les effets indépendants des différentes composantes du traitement et d’évaluer les effets du programme sur davantage de pratiques parentales associées spécifiquement à l’anxiété. / Separation anxiety disorder (SAD) is the most prevalent anxiety disorder among children. It appears early in development and has multiple negative consequences. Cognitive-behavioral therapy (CBT) has been shown to be an effective treatment for anxiety disorders. However, few studies have examined the efficacy of CBT to treat SAD in particular, and even fewer have examined the impact of this form of therapy on children younger than 7 years old. The main objective of the present doctoral thesis was to evaluate the efficacy of a CBT parent-training program, that was adapted specifically for parents of children aged 4 to 7 years old suffering from SAD. This study had two specific objectives: to observe any fluctuations in the child’s SAD symptoms during the treatment and to examine the impact of the program on parental variables. A single-case multiple baseline across-subjects design was used. Six families with a child aged 4-7 years old and with a diagnosis of SAD participated. Semi-structured interviews, self-reported questionnaires and daily diaries were used with the parents to assess the child’s anxiety symptoms, parental practices, parenting stress, and the parents anxious and/or depressive symptoms. Questionnaires on child problems were also sent to the children’ teacher or educator. All questionnaires were administered at three times of measurement (pre-treatment, post-treatment and 3 months follow-up). Daily diaries were also completed by the parents at baseline, throughout the treatment, and during one to two weeks at follow-up. Results revealed that five of the six children no longer met the criteria of a SAD diagnosis after treatment and three months later. Findings from the daily diaries showed a clear reduction of the principal SAD symptoms for half of the children but mixed results for the other half of the children and that four of six children presented a systematic favourable change of the total weekly frequency of SAD symptoms after the intervention. The results of parent questionnaires showed an improvement of SAD symptoms for four children at post-treatment and/or follow-up. The teacher/educator questionnaires indicated low impact of anxiety symptoms. The impact of the program on parenting stress and parental practices is mixed with some parents showing improvement and others less so. The results support the efficacy of the Programme d’entraînement parental pour les enfants souffrant d’anxiété de séparation (PEP-AS) to reduce SAD symptoms in preschool age children and support the relevance of directing the treatment towards parents and including a relational component in the intervention. However, more research is needed to replicate these findings with larger samples and randomized control trials. It would also be interesting to dismantle the program and to examine the various components of the treatment in different combinations, and to explore more specifically the program effects on parental variables.

Made available in DSpace on 2016-04-04T18:22:08Z (GMT). No. of bitstreams: 1 Renan Amauri Guaranha Rinaldi.pdf: 5019609 bytes, checksum: 6777801213f438ccb3f67ed4e7802152 (MD5) Previous issue date: 2014-02-14 / In 1549, after the approval of Jesus? Society in Rome, the P. Manuel da N?brega, left Lisbon with the armed Tom? de Souza to conduct the West Portuguese missions. In 1553, after forming the first Brazilian missions Ba?a de Todos os Santos where they landed, he continued to the Captainship of S?o Vicente where he covered the south coast of colonial Brazil. After that, going up through Serra do Mar, N?brega lead the foundation of Aldeia de Piratininga and its school, Col?gio de S?o Paulo, and posteriorly, he enabled the beginning of new entries to the interior of the continent. The implementation of these schools originated the villages or urban areas such as Vila de S?o Paulo, Rio de Janeiro and Vit?ria, afterwards becoming centers of irradiation of colonial expansion forming small network localities. The sites related to these schools and Jesuit? churches and settlements designed to reduce Indian population formed the architectonic sites of the settlements, while the organization of the lodges focused to give support to evangelization and the introduction of Indians to Portuguese life system. This work intends to analyze the acting of Jesus? Society according to the purpose of Portuguese colonizers during the XVI century until half of XVII century, before the pombal political expulsion, and the logic of expansion in the Captainships in the south of Brazilian territory in the colonial period, focusing on territorial expansion accomplished after the influence of Col?gio de S?o Paulo, and Vila de S?o Jos? dos Campos in Vale do Parna?ba and Fazenda de Botucatu in the west side of the Captainship as being the last two stronghold of Jesuit? presence in the territory of colonial period. To accomplish that there will be a support on historiography and analysis of Jesuits letters besides other studies about the rise and expansion of the settlements based on antique and contemporary cartographies demonstrating the presence of Jesuits in the mentioned territory. / Em 1549, logo ap?s a aprova??o da Companhia de Jesus em Roma, o P. Manuel da N?brega partiu de Lisboa na armada de Tom? de Souza juntamente de cinco companheiros para conduzir as Miss?es Portuguesas do Ocidente. Em 1553, ap?s formar as primeiras miss?es brasileiras na Ba?a de Todos os Santos, onde primeiramente aportou, seguiu em dire??o ? Capitania de S?o Vicente onde percorreu o litoral sul do Brasil colonial. Em seguida, subindo a Serra do Mar, N?brega liderou a funda??o da Aldeia de Piratininga e nela implantou o Col?gio de S?o Paulo possibilitando o in?cio das entradas para o interior do continente. As implanta??es destes col?gios por vezes deram origem a n?cleos urbanos, como nos casos das Vilas de S?o Paulo, Rio de Janeiro e Vit?ria, que se tornaram posteriormente centros de irradia??o da expans?o colonial formando redes de pequenas localidades. Os p?tios e terreiros associados a esses col?gios, bem como as igrejas jesu?ticas e os aldeamentos destinados ? redu??o ind?gena formavam os conjuntos arquitet?nicos dos aldeamentos enquanto a organiza??o do espa?o das resid?ncias visava dar suporte ? evangeliza??o e a introdu??o do ?ndio ao sistema de vida portugu?s. Este trabalho tem como prop?sito analisar a atua??o da Companhia de Jesus perante os prop?sitos dos colonizadores portugueses durante o s?culo XVI at? primeira metade do s?culo XVIII, antes da pol?tica pombalina de expuls?o, e a l?gica de expans?o nas Capitanias ao sul do territ?rio brasileiro no per?odo colonial, concentrando o olhar na expans?o territorial realizada a partir da influ?ncia do Col?gio de S?o Paulo, sendo a Vila de S?o Jos? dos Campos no Vale do Para?ba e a Fazenda de Botucatu no oeste da Capitania, os dois ?ltimos redutos da presen?a jesu?tica no territ?rio do per?odo colonial. Para tal, nos apoiaremos na historiografia e na an?lise de cartas jesu?ticas, al?m de estudos sobre os fluxos e expans?o dos assentamentos tendo como base as cartografias antiga e contempor?nea, demonstrando a presen?a dos jesu?tas no territ?rio em quest?o.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-04-17T18:46:28Z No. of bitstreams: 1 467348 - Texto Completo.pdf: 1745226 bytes, checksum: 1e33ae6db8765aeb6f854daed1c08f43 (MD5) / Made available in DSpace on 2015-04-17T18:46:28Z (GMT). No. of bitstreams: 1 467348 - Texto Completo.pdf: 1745226 bytes, checksum: 1e33ae6db8765aeb6f854daed1c08f43 (MD5) Previous issue date: 2015-03-16 / This study aims to analyze the foundation of Vila Maria of Paraguay, current C?ceres - MT, with the presence of Chiquitano Indians from Chiquitano Jesuit missions on the eastern side of the Spanish colony in America, now Bolivia, probably, Santa Ana, Santa Rosa, St. John and the Holy Heart of Jesus de Chiquitos. Seeks to highlight the Portuguese settlement strategies with these Indians in the period they were created and consolidated the Portuguese-Spanish border in western Mato Grosso Captaincy with the lands of the Spanish colony in South America especially in Vila Maria of Paraguay. The time frame covers the foundation of Vila Maria from 1778 to 1827, during the view of Hercules Florence to that village. Vila Maria becomes important strategic point between the villages of Cuiab? and Vila Bela of the Trinity in the Guapor? valley and the lack of "white people" to populate it, the Chiquitano and Indians from other ethnic groups, served the Lusitanian project of settlement, occupation and consolidation of their land in this part of the colony. The royal instructions and the urbanization project of the Portuguese crown and the Marquis of Pombal will be the hallmarks of this settlement. / Este estudo tem como objetivo analisar a funda??o de Vila Maria do Paraguai, atual C?ceres ? MT, com a presen?a de ?ndios Chiquitano provenientes de miss?es jesu?ticas Chiquitanas do lado oriental da col?nia espanhola na Am?rica, atual Bol?via, provavelmente de, Santa Ana, Santa Rosa, S?o Jo?o e do Santo Cora??o de Jesus de Chiquitos. Procura evidenciar as estrat?gias de povoamento dos portugueses com esses ?ndios no per?odo em que se criavam e consolidavam a fronteira luso-espanhola no oeste da Capitania de Mato Grosso com as terras da col?nia espanhola na Am?rica do sul, especialmente em Vila Maria do Paraguai. O recorte temporal abrange a funda??o de Vila Maria 1778 a 1827, ocasi?o da vista de Hercules de Florence a essa Vila. Vila Maria torna-se importante ponto estrat?gico entre as Vilas de Cuiab? e Vila Bela da Sant?ssima Trindade no vale do Guapor? e na falta de ?gente branca? para povo?-la, os Chiquitano, bem como ?ndios de outras etnias, serviram ao projeto lusitano de povoamento, ocupa??o e consolida??o de suas terras nesta parte da col?nia. As instru??es r?gias e o projeto de urbaniza??o da coroa portuguesa e do Marqu?s de Pombal ser?o os balizadores deste povoamento.

<p>The analysis of low abundant biological molecules is often challenging due to their chemical properties, low concentration and limited sample volumes. Neuropeptides are one group of molecules that fits these criteria. Neuropeptides also play an important role in biological functions, which makes them extra interesting to analyze. A classic chemical analysis involves sampling, sample preparation, separation and detection. In this thesis, an enhanced solid supported microdialysis method was developed and used as a combined sampling- and preparation technique. In general, significantly increased extraction efficiency was obtained for all studied peptides. To be able to control the small sample volumes and to minimize the loss of neuropeptides because of unwanted adsorption onto surfaces, the subsequent analysis steps were miniaturized to a micro total analysis system (µ-TAS), which allowed sample pre-treatment, injection, separation, manipulation and detection. </p><p>In order to incorporate these analysis functions to a microchip, a novel microfabrication protocol was developed. This method facilitated three-dimensional structures to be fabricated without the need of clean room facilities. </p><p>The sample pre-treatment step was carried out by solid phase extraction from beads packed in the microchip. Femtomole levels of neuropeptides were detected from samples possessing the same properties as microdialysates. The developed injection system made it possible to conduct injections from a liquid chromatographic separation into a capillary electrophoresis channel, which facilitated for advanced multidimensional separations. An electrochemical sample manipulation system was also developed. In the last part, different electrospray emitter tip designs made directly from the edge of the microchip substrate were developed and evaluated. The emitters were proven to be comparable with conventional, capillary based emitters in stability, durability and dynamic flow range. Although additional developments remain, the analysis steps described in this thesis open a door to an integrated, on-line µ-TAS for neuropeptides analysis in complex biological samples.</p>

The analysis of low abundant biological molecules is often challenging due to their chemical properties, low concentration and limited sample volumes. Neuropeptides are one group of molecules that fits these criteria. Neuropeptides also play an important role in biological functions, which makes them extra interesting to analyze. A classic chemical analysis involves sampling, sample preparation, separation and detection. In this thesis, an enhanced solid supported microdialysis method was developed and used as a combined sampling- and preparation technique. In general, significantly increased extraction efficiency was obtained for all studied peptides. To be able to control the small sample volumes and to minimize the loss of neuropeptides because of unwanted adsorption onto surfaces, the subsequent analysis steps were miniaturized to a micro total analysis system (µ-TAS), which allowed sample pre-treatment, injection, separation, manipulation and detection. In order to incorporate these analysis functions to a microchip, a novel microfabrication protocol was developed. This method facilitated three-dimensional structures to be fabricated without the need of clean room facilities. The sample pre-treatment step was carried out by solid phase extraction from beads packed in the microchip. Femtomole levels of neuropeptides were detected from samples possessing the same properties as microdialysates. The developed injection system made it possible to conduct injections from a liquid chromatographic separation into a capillary electrophoresis channel, which facilitated for advanced multidimensional separations. An electrochemical sample manipulation system was also developed. In the last part, different electrospray emitter tip designs made directly from the edge of the microchip substrate were developed and evaluated. The emitters were proven to be comparable with conventional, capillary based emitters in stability, durability and dynamic flow range. Although additional developments remain, the analysis steps described in this thesis open a door to an integrated, on-line µ-TAS for neuropeptides analysis in complex biological samples.

Abstract Somatization is a widespread phenomenon causing subjective suffering and disability. The aim of the study was to assess somatization disorder (SD) and somatization symptoms among young adult population and their associations with sociodemographic factors, alexithymia and temperament as well as psychiatric comorbidity. Various suggestions have been presented to operationalize somatization but none of them has been shown to be superior to others. In this study two definitions were used: SD by DSM-III-R classification diagnostic criteria and "somatization" meaning four or more symptoms of the 35 symptoms of DSM-III-R SD criteria. The study population was a subsample of the Northern Finland Birth Cohort 1966 (NFBC 1966), consisting of cohort members living in Oulu (N = 1,609) on January 1st 1997. The NFBC 1966 is a general population birth cohort of 12,058 live-born children covering 96.3% of all deliveries in the catchment area. The best-estimated procedure was used for assessment of psychiatric morbidity including SD and somatization. Data were collected from the Finnish Hospital Discharge Register and from all available outpatient and inpatient records. Data on education were gathered from Statistics Finland. Other sociodemographic variables, alexithymia and temperament scores were drawn from questionnaires of the field study conducted in 1997 and from earlier follow-up studies. The prevalence of SD was 1.1% (N = 18). Of the subjects 6.1% (N = 97) had somatization. The female-to-male ratio was 5:1 and 6:1, respectively. SD was not recognized by any of the treating physicians, at least not documented in case notes. The observed occurrences of SD and somatization were at a level comparable with earlier international population studies. Somatization did not associate with depression or alexithymia, and neither could a characteristic temperament profile be recognized. Somatization was associated with psychological distress. These results indicate a need for training physicians to recognize SD and somatization and its comorbidity. This will have implications both for psychiatry and other medical specialties regarding collaboration and underlines the importance of liaison-psychiatry at general hospitals. The results suggest a need for more studies about the etiology and development of SD and somatization. / Tiivistelmä Somatisaatio on yleinen ilmiö, josta aiheutuu subjektiivista kärsimystä ja toimintakyvyn laskua. Tämän tutkimuksen tarkoitus oli arvioida somatisaatiohäiriön ja somatisaatio-oireilun yleisyyttä nuorilla aikuisilla sekä näiden ilmiöiden yhteyttä sosiodemografisiin tekijöihin, aleksitymiaan, temperamenttiin ja psykiatriseen sairastavuuteen. Somatisaation käsitteellistämiseksi on esitetty useita vaihtoehtoja mutta mikään niistä ei ole osoittautunut muita paremmaksi. Tässä tutkimuksessa käytetiin kahta määritelmää: DSM-III-R -diagnoosiluokituksen mukaista somatisaatiohäiriön diagnoosia tai somatisaatio-oireilua, jossa esiintyy neljä tai useampia DSM-III-R:n 35 somatisaatiohäiriön oireesta. Tutkimusaineiston muodostivat Pohjois-Suomen vuoden 1966 syntymäkohortin ne jäsenet, jotka asuivat Oulussa 1. tammikuuta 1997 (N =  1,609). Alkuperäinen kohortti koostuu 12,058 elävänä syntyneestä tutkittavasta, mikä kattaa 96.3 % kaikista synnytyksistä Pohjois-Suomessa. Niin kutsutun best-estimated -menettelyn avulla arvioitiin tutkittavien psykiatrista sairastavuutta mukaan lukien somatisaatiohäiriö ja -oireilu. Tietoa kerättiin sairaaloiden poistoilmoitusrekisteristä. Avohoidon sairauskertomustieto koottiin kattavasti. Koulutusasteesta saatiin tieto Tilastokeskukselta. Muita sosiodemografisia tekijöitä, aleksitymiaa ja temperamenttia arvioitiin vuoden 1997 kenttätutkimuksen ja aiempien seurantatutkimusten tietojen avulla. Somatisaatiohäiriön esiintyvyys oli 1.1 % (N =  18). Somatisaatio-oireita todettiin 6.1 % (N =  97) tutkittavista. Naisten osuus oli somatisaatiohäiriössä 5:1 ja somatisaatio-oireilussa 6:1. Osoittautui, että lääkärit eivät tunnistaneet somatisaatiohäiriötä, ainakaan sitä ei oltu kirjattu sairauskertomuksiin. Havaitut somatisaatiohäiriön ja -oireilun esiintyvyydet ovat sopusoinnussa aiempien kansainvälisten tutkimusten kanssa. Somatisaatio-oireilu ei liittynyt masennukseen tai aleksitymiaan eikä somatisaatio-oireilusta kärsiville tutkittavilla todettu tyypillistä temperamenttiprofiilia. Somatisaatio liittyi psyykkiseen stressiin. Johtopäätöksenä voidaan todeta, että lääkäreille tulisi tarjota koulutusta somatisaatiohäiriön ja -oireilun tunnistamisessa. On tärkeää tunnistaa somatisaatio ja siihen liittyvä oheissairastavuus. Havainnot korostavat yleissairaaloiden yhteistyöpsykiatrian ja muiden erikoisalojen yhteistyön merkitystä somatisaatiosta kärsivien potilaiden tutkimuksessa ja hoidossa. Somatisaatiohäiriön ja -oireilun etiologian ja kehittymisen selvittämiseksi tarvitaan uusia tutkimuksia.