Introdução: As Doenças lisossômicas (DLs) são condições genéticas, herdadas na sua maioria de forma autossômica recessiva, caracterizadas usualmente pela deficiência de enzimas lisossômicas específicas, envolvidas na síntese, degradação, armazenamento ou transporte de macromoléculas necessárias para o funcionamento normal do organismo. Nas situações mais típicas, o substrato não degradado acumula-se progressivamente nos lisossomos, com repercussões estruturais e funcionais, levando a sinais e sintomas característicos. Os pacientes apresentam um amplo espectro de manifestações clínicas, que podem incluir disfunção de órgãos, anormalidades esqueléticas, envolvimento neuronal, entre outras. O diagnóstico é usualmente obtido pela identificação da deficiência enzimática específica em leucócitos obtidos do sangue periférico, usualmente realizado em laboratórios de referência. O transporte da amostra pode ser um obstáculo quando o serviço requisitante está situado longe do centro de referência ou em outro país, situação em que a amostra de sangue pode chegar ao laboratório já sem condições de ser analisada. Objetivo: Este estudo teve como objetivo principal, tornar disponível um método mais simples, seguro e acessível que utiliza amostras de leucócitos impregnados em papel filtro (LIPF) como uma nova ferramenta para o diagnóstico bioquímico de pacientes com DLs. Métodos: O estudo envolveu amostras de pacientes com diagnóstico previamente confirmado de DLs (amostra de conveniência, por se tratarem de doenças raras, com incidências individuais ao redor de 1:100.000 recém-nascidos vivos). Foram incluídos no estudo os pacientes com diagnóstico já estabelecido de DLs selecionadas (MPS IVA, Doença de Krabbe, Doença de Gaucher e Doença de Pompe), independente do sexo e/ou idade, atendidos no Serviço de Genética Médica do HCPA, que concordaram em participar do estudo. O grupo de referência negativo foi constituído pelas amostras de 50 indivíduos hígidos, adultos, de ambos os sexos. Resultados: Os resultados obtidos nos ensaios enzimáticos de pacientes com MPS IVA, Doença de Krabbe, Doença de Gaucher e Doença de Pompe, indicaram que as enzimas analisadas em amostras de LIPF permitiram a identificação de todos os pacientes, com sensibilidade de 100%. Os testes de estabilidade realizados nas amostras de LIPF indicaram que as amostras, quando mantidas a 4ºC, se mostram estáveis por pelo menos 30 dias. Conclusões: Nas condições utilizadas, amostras de LIPF se mostraram adequadas para a identificação segura de pacientes com MPS tipo IVA, Doença de Krabbe, Doença de Gaucher e Doença de Pompe. As amostras de leucócitos secos em papel filtro são mais estáveis e seguras para o transporte, indicando que possa ser esta uma importante ferramenta para facilitar a identificação de pacientes com DLDs, especialmente daqueles que vivem em áreas que tem dificuldades para a remessa de amostras líquidas para serviços de referência. / Background: Lysosomal Disorders (LDs) are genetic conditions, mostly inherited in autosomal recessive fashion, usually characterized by a deficiency of specific lysosomal enzymes involved in the synthesis, degradation, storage or transportation of macromolecules necessary for normal functioning of the organism. Typically, the non-degraded substrate is progressively accumulated in lysosomes, with structural and functional repercussions, leading to characteristic signs and symptoms. Affected patients present a wide range of clinical manifestations, which may include organ dysfunction, skeletal anomalies, neuronal involvement, etc. The diagnosis is normally made through identification of the specific enzyme deficiency in white blood cells from a sample of peripheral blood, usually performed in reference laboratories. The transporting of a liquid sample can be a problem when the test orderer is located far from the reference center or in a foreign country, as often the blood sample arrives at the laboratory in poor condition and cannot be properly analyzed. Aim: The main aim of this study was to make available a new technique that is simpler, safer and more accessible, using leukocytes impregnated on filter paper (LIFP) as a new tool for the biochemical diagnosis of patients with LSDs. Methods: This study involved samples of patients with previously confirmed diagnosis of selected LSDs (a convenience sample, as these are rare diseases, with individual incidences around 1:100.000 live newborns). Patients with an established diagnosis of MPS IVA, Krabbe Disease, Gaucher’s disease and Pompe disease regardless of sex and/or age, cared for at the Genetics Service of HCPA and who agreed to participate were included in the study. The negative reference group comprised blood samples from 50 healthy adults of both genders. Results: The results obtained in the enzymatic assays of patients with MPS IVA, Krabbe Disease, Gaucher’s disease, and Pompe disease indicated that the analyzed enzymes in LIFP samples allowed the identification of all patients, with sensitivity of 100%. The stability tests performed in LIFP samples indicated that samples, when maintained at 4ºC, were stable for at least 30 days. Conclusions: In the conditions used, LIFP samples were shown to be adequate for a reliable identification of patients with MPS IVA, Krabbe Disease, Gaucher’s disease, and Pompe disease. Blood samples on filter paper are more stable and reliable for transportation, indicating that this may be an important tool to facilitate the identification of patients with LSDs, particularly those living in areas with difficulties for the shipment of liquid samples to reference cervices.

This master's thesis concerns itself with the motive of "despair" in contemporary Czech drama. It includes an analysis of selected plays written after 2000 by leading Czech authors belonging to the middle generation (Petr Zelenka; David Drábek; Jiří Pokorný; Petr Kolečko; Tomáš Svoboda; Lenka Lagronová; Iva Klestilová, born Volánková). The aim of the thesis is to analyze these dramas, to define characteristic elements and to determine whether we can identify a clear "desperate people" motive within their characters, deeming it a characteristic feature of contemporary Czech drama written by the middle generation. Using a comparative method we explore typical repetitive motives of the characters - anti- heroes, which suffer from having an unfulfilled personal life, a feeling of loneliness or a midlife crisis. The master's thesis describes the causes and manifestations of despair of the characters, as well as the solutions the dramas offer to the bleak situation (insanity, suicide, miracle). KEYWORDS contemporary Czech drama, motive of "losers", in-yer-face theatre, Petr Zelenka, David Drábek, Jiří Pokorný, Petr Kolečko, Tomáš Svoboda, Lenka Lagronová, Iva Klestilová (Volánková)

Introdução: As Doenças lisossômicas (DLs) são condições genéticas, herdadas na sua maioria de forma autossômica recessiva, caracterizadas usualmente pela deficiência de enzimas lisossômicas específicas, envolvidas na síntese, degradação, armazenamento ou transporte de macromoléculas necessárias para o funcionamento normal do organismo. Nas situações mais típicas, o substrato não degradado acumula-se progressivamente nos lisossomos, com repercussões estruturais e funcionais, levando a sinais e sintomas característicos. Os pacientes apresentam um amplo espectro de manifestações clínicas, que podem incluir disfunção de órgãos, anormalidades esqueléticas, envolvimento neuronal, entre outras. O diagnóstico é usualmente obtido pela identificação da deficiência enzimática específica em leucócitos obtidos do sangue periférico, usualmente realizado em laboratórios de referência. O transporte da amostra pode ser um obstáculo quando o serviço requisitante está situado longe do centro de referência ou em outro país, situação em que a amostra de sangue pode chegar ao laboratório já sem condições de ser analisada. Objetivo: Este estudo teve como objetivo principal, tornar disponível um método mais simples, seguro e acessível que utiliza amostras de leucócitos impregnados em papel filtro (LIPF) como uma nova ferramenta para o diagnóstico bioquímico de pacientes com DLs. Métodos: O estudo envolveu amostras de pacientes com diagnóstico previamente confirmado de DLs (amostra de conveniência, por se tratarem de doenças raras, com incidências individuais ao redor de 1:100.000 recém-nascidos vivos). Foram incluídos no estudo os pacientes com diagnóstico já estabelecido de DLs selecionadas (MPS IVA, Doença de Krabbe, Doença de Gaucher e Doença de Pompe), independente do sexo e/ou idade, atendidos no Serviço de Genética Médica do HCPA, que concordaram em participar do estudo. O grupo de referência negativo foi constituído pelas amostras de 50 indivíduos hígidos, adultos, de ambos os sexos. Resultados: Os resultados obtidos nos ensaios enzimáticos de pacientes com MPS IVA, Doença de Krabbe, Doença de Gaucher e Doença de Pompe, indicaram que as enzimas analisadas em amostras de LIPF permitiram a identificação de todos os pacientes, com sensibilidade de 100%. Os testes de estabilidade realizados nas amostras de LIPF indicaram que as amostras, quando mantidas a 4ºC, se mostram estáveis por pelo menos 30 dias. Conclusões: Nas condições utilizadas, amostras de LIPF se mostraram adequadas para a identificação segura de pacientes com MPS tipo IVA, Doença de Krabbe, Doença de Gaucher e Doença de Pompe. As amostras de leucócitos secos em papel filtro são mais estáveis e seguras para o transporte, indicando que possa ser esta uma importante ferramenta para facilitar a identificação de pacientes com DLDs, especialmente daqueles que vivem em áreas que tem dificuldades para a remessa de amostras líquidas para serviços de referência. / Background: Lysosomal Disorders (LDs) are genetic conditions, mostly inherited in autosomal recessive fashion, usually characterized by a deficiency of specific lysosomal enzymes involved in the synthesis, degradation, storage or transportation of macromolecules necessary for normal functioning of the organism. Typically, the non-degraded substrate is progressively accumulated in lysosomes, with structural and functional repercussions, leading to characteristic signs and symptoms. Affected patients present a wide range of clinical manifestations, which may include organ dysfunction, skeletal anomalies, neuronal involvement, etc. The diagnosis is normally made through identification of the specific enzyme deficiency in white blood cells from a sample of peripheral blood, usually performed in reference laboratories. The transporting of a liquid sample can be a problem when the test orderer is located far from the reference center or in a foreign country, as often the blood sample arrives at the laboratory in poor condition and cannot be properly analyzed. Aim: The main aim of this study was to make available a new technique that is simpler, safer and more accessible, using leukocytes impregnated on filter paper (LIFP) as a new tool for the biochemical diagnosis of patients with LSDs. Methods: This study involved samples of patients with previously confirmed diagnosis of selected LSDs (a convenience sample, as these are rare diseases, with individual incidences around 1:100.000 live newborns). Patients with an established diagnosis of MPS IVA, Krabbe Disease, Gaucher’s disease and Pompe disease regardless of sex and/or age, cared for at the Genetics Service of HCPA and who agreed to participate were included in the study. The negative reference group comprised blood samples from 50 healthy adults of both genders. Results: The results obtained in the enzymatic assays of patients with MPS IVA, Krabbe Disease, Gaucher’s disease, and Pompe disease indicated that the analyzed enzymes in LIFP samples allowed the identification of all patients, with sensitivity of 100%. The stability tests performed in LIFP samples indicated that samples, when maintained at 4ºC, were stable for at least 30 days. Conclusions: In the conditions used, LIFP samples were shown to be adequate for a reliable identification of patients with MPS IVA, Krabbe Disease, Gaucher’s disease, and Pompe disease. Blood samples on filter paper are more stable and reliable for transportation, indicating that this may be an important tool to facilitate the identification of patients with LSDs, particularly those living in areas with difficulties for the shipment of liquid samples to reference cervices.

Bakgrund   The Gugging Swallowing Screen - Intensive Care Unit (GUSS-ICU) är ett screeningtest med syfte att fånga upp indikationer på sväljsvårigheter efter långvarig intubering hos inneliggande patienter på intensivvårdsavdelningar (IVA). I dagsläget finns det inget svenskt översatt och validerat screeningtest för omvårdnadspersonalen att använda vid bedömning av sväljförmågan efter extubering på IVA. Syfte Syftet med studien var att översätta det internationella screeningtestet GUSS-ICU till svenska för användning i en svensk intensivvårdspopulation. Vidare syftade studien till att utföra en pilotstudie av den svenska versionen på extuberade patienter på IVA. Metod En framåt-bakåtöversättningsmetod användes vid översättningen av screeningtestet GUSS-ICU. Översättningsprocessen omfattade tre steg: framåtöversättning, granskning och kommentarer från en expertpanel och en bakåtöversättning. I översättningsprocessen deltog två logopedstudenter, två handledare, en expertgrupp och en översättare. Expertgruppen bestod av 10 deltagare med olika professioner inom hälso- och sjukvården. Fem inneliggande patienter från IVA planerades delta i en pilottestning av den svenska översättningsversionen. En innehållsanalys genomfördes för att kunna jämföra samtliga översättningar. Syftet med jämförelsen var att identifiera skillnader i ord och satser samt kulturella och kontextuella skillnader. Resultat       Studien resulterade i en svensk översättning (GUSS-IVA) av screeningtestet GUSS-ICU. Analysen visade på skillnader i val av ord, koncept och satser mellan de olika översättningarna. Flertalet skillnader var ej betydelseskiljande utan analyserades istället som resultat av skillnader i erfarenhet, kunskap och språkbruk. Översättningsmetoden bidrog till språklig, kontextuell och kulturell anpassning av översättningen. Slutsats Den svenska versionen av GUSS-ICU stämmer bra överens med originalversionen och enbart ett fåtal skillnader observerades mellan översättningarna. Skillnaderna mellan bakåtöversättningen och originalversionen var inte betydelseskiljande, vilket tyder på att den svenska versionen mäter det den avser att mäta. Framtida studier behöver pilottesta och validera den svenska versionen av GUSS-ICU, innan testet kan implementeras i en svensk intensivvårdspopulation. / Validering av screeningtest för sväljsvårigheter för användning inom svensk vårdkontext

Mucopolysaccharidoses are a group of diseases that belong to lysosomal storage disorders. A common sign of these monogenic multisystem diseases is a gene mutation leading to a deficiency of the lysosomal enzyme participating in glycosaminoglycan degradation. It results to their accumulation in the tissues and organs, where they cause a progressive damage. There is no efficient treatment available for most mucopolysaccharidoses. Moreover, the research is complicated because of the low prevalence and type of affected tissues. Animal models of these human diseases are used for an evaluation of newly developed therapeutic approaches. However, they also have many limitations due to the different pathogenesis and catabolic pathways of the accumulated substrates between humans and animals. Therefore, animal models are replaced by human cell models. In this thesis, the development of four mucopolysaccharidoses human cell models is reported (MPS IIID, MPS IVA, MPS IVB, MPS VI). Corresponding genes (GNS, GALNS, GLB1, ARSB) were inactivated using CRISPR/Cas9 technology, where plasmids containing specific inserts are delivered to the target human induced pluripotent stem cells (iPSC), using electroporation. Isolated clones, which represent iPSC disease models, were characterized by Sanger sequencing, enzyme...

Bakgrund: Specialistsjuksköterskor med inriktning mot intensivvård har ansvar för omvårdnaden av de svårast sjuka patienterna samt kommunicerar med deras närstående. Coronaviruset (SARS-CoV-2) även kallad för Covid-19, leder till en atypisk form av akut andningsbesvär som i vissa fall kräver ventilatorbehandling. Dessa patienter kommer alla att behöva urträning ur respiratorn inför extubation. Urträning initieras oftast på intensivvårdsavdelningen och tillhör intensivvårdssjuksköterskans arbete. En förkortad ventilatorbehandling kan minska patientens lidande, mortaliteten och sänka samhällskostnader. Syfte: Syftet med studien var att beskriva intensivvårdssjuksköterskors framgångsfaktorer för ventilatorurträning av patienter med Covid-19. Metod: Vi har använt oss av kvalitativ metod och intervjuat åtta intensivvårdssjuksköterskor i Västra Sverige. Intervjuerna analyserades kvalitativ innehållsanalys med manifest ansats som resulterade i tre kategorier. Resultat: Denna kvalitativa intervjustudie visade att en längre och svårare urträningsprocess kräver tålamod och försiktighet. Samtliga intensivvårdssjuksköterskor beskrev urträningsprocessen för patienter med Covid-19 som både längre och svårare i jämförelse med patienter som ventilatorbehandlats av andra orsaker. Tidiga insatser kan lindra sjukdomsförlopp och underlätta urträningen. Intensivvårdssjuksköterskorna beskrev att de under pandemin lärt sig använda både medicinska och omvårdnadsmässiga insatser tidigt för att bromsa, lindra eller förhindra ett progrederande sjukdomsförlopp. Mobilisering, teamsamverkan och kontinuitet var viktigt vid urträning och framgångsfaktorer som påverkades av personalens kompetens, stressnivå och andraorganisatoriska faktorer. Slutsats: Denna kvalitativa intervjustudie visade att intensivvårdssjuksköterskor upplever att erfarenhet under pandemins gång har förbättrat förutsättningarna för patienter med Covid-19 i behovav urträning från ventilatorbehandling. Och dessa lärdomar ligger till grund för de faktorer som i dagsläget ofta ligger till grund för en framgångsrik ventilatorurträning. Deras erfarenhet och kunskaper skiljer sig åt och resultatet tyder på att mer utbildning och kontinuitet krävs för att kunna ge en säkrare vård.

Patienter som vårdats på en intensivvårdsavdelning (IVA) och tillfrisknat efter kritisk sjukdom löper stor risk för komplikationer efter att de flyttats till annan vårdavdelning för fortsatt vård. Oförutsedda komplikationer kan ibland leda det till att patienten återinläggs på IVA. Tidig identifiering och nära samarbete mellan IVA och vårdavdelning kring vanliga komplikationer skulle kunna förbättra förutsättningar för effektiv rehabilitering för den enskilda patienten.   Syftet med förbättringsarbetet var att underlätta patientens rehabilitering genom att tillgodose dennes behov av information och specifika rehabiliteringsinsatser som är vanliga efter intensivvård. återhämtning efter intensivvård samt minska risken för återinläggning.    Under förbättringsarbetet infördes ett screeningsinstrument för att tidigt kunna fånga patientens behov av stöd. Förbättringsarbetet utgick från Nolans förbättringsmodell. Resultatet av förbättringsarbetet visar på att det finns behov av tidigare uppföljning av intensivvårdspatienten för att förbättra och stötta patienten i sin återhämtning. Screeningsinstrumentet bidrog till att de patienter som är i behov av stöd fångades upp.    Kunskap om utmaningar och framgångsfaktorer under förbättringsarbete kan bidra till en större förståelse för förbättringskunskapens betydelse vid utveckling av vården. Syftet med studien var att belysa intensivvårdspersonalens erfarenheter av förbättringsarbete inom post-intensivvård.   En kvalitativ studie genomfördes med två fokusgruppsintervjuer bestående av vårdpersonal från en IVA-avdelning. Under intervjuerna fångades vårdpersonalens upplevelse av förbättringsarbete. De transkriberade intervjuerna analyserades med en kvalitativ innehållsanalys. I resultatet av studien framkom både utmaningar och framgångsfaktorer i tre kategorier: ”Skapa mening och förståelse för nytt arbetssätt”, ”Proaktivt arbete med patientfokus” samt ”Förändringströtthet och brist på tid”.   Tidigare uppföljning är en viktig del i patientens återhämtning efter IVA-vård för att förebygga senkomplikationer. Studien visar att tydlig information kring interventioner bidrar till ökad förståelse för förändringsprocessen och skapar drivkraft i förbättringsarbetet. / Patients who recovers from severe illness entails an increased risk of complications when being transferred from intensive care unit (ICU) to ward. Unforeseen complications can sometimes lead to ICU re-admissions. Early identification of common complications and close collaboration between ICU and ward can improve the chances of a successful recovery for the patient.   The purpose of the improvement work was to facilitate the patient's recovery after intensive care and to reduce the risk of readmission. rehabilitation by meeting his or her need for information and specific rehabilitation interventions that are common after intensive care.   During the improvement work, a screening instrument predicting high risk-individuals was introduced to capture the patient's need for support at an early stage. The improvement work was based on Nolan's improvement model. The results of the improvement work showed that there is a need for earlier follow-up of the ICU-patient to improve and support patients in their recovery. The screening instrument helped to catch the patients in need of support.   Knowledge of challenges and success factors during improvement work can contribute to a greater understanding of the importance of improvement knowledge in the developement of care. The purpose of the study was to illuminate intensive care staff's experiences of improvement work in post-intensive care.   A qualitative study was conducted with two focus group interviews consisting of care staff from an ICU department. During the interviews, the care staff's experience of improvement work was captured. The transcribed interviews were analyzed with a qualitative content analysis. Focus group interviews with staff resulted in three categories: "Creat meaning and understanding for a new way of working", "Proactive work with a patient focus" and "Change fatigue and lack of time".  Previous follow-up is an important part of the patient's recovery to detect late complications. The study result concludes that clear information according to interventions during improvement work contributes to increased understanding for the improvement process and creates motivation and drive in the improvement work.

Questa tesi include tre capitoli che si concentrano sulle politiche pubbliche e la tutela della salute. Il primo capitolo esamina l’effetto dell’incremento delle aliquote dell’IVA su alcune bevande sul loro consumo. I risultati dell’analisi empirica suggeriscono che l'aumento dell'aliquota IVA ha ridotto il consumo di alcolici, birra e bevande gassate, mentre l’effetto sul consumo di vino non è statisticamente significativo. Tuttavia, questo risultato generale cambia se si considerano sotto-gruppi di individui. Il secondo capitolo studia l’impatto del salario minimo sulla salute della popolazione cinese. Le stime indicano che i salari minimi reali sono negativamente e significativamente correlati con lo stato di salute della popolazione sia nel breve che nel lungo periodo, un risultato che potrebbe essere spiegato dalle condizioni di lavoro più stressanti conseguenti all’introduzione di un salario minimo più elevato. Infine, il terzo capitolo analizza gli effetti dei meccanismi di incentivazione sui comportamenti degli operatori sanitari del settore delle cure primarie nell’ambito della gestione dell'ipertensione. La nostra indagine empirica suggerisce che gli incentivi finanziari negativi potrebbero motivare gli operatori sanitari a migliorare la qualità dei servizi legati alla gestione dell'ipertensione, mentre né gli incentivi finanziari né diverse modalità di valutazione delle prestazioni sembrano influenzare la qualità dei servizi. / This dissertation includes three chapters which focus on the relationship between public policies and healthcare. The first chapter examines whether tax policy is effective in changing the unhealthy drinking behaviours of individuals. My findings suggest that the VAT rate increase is effective in reducing the consumption of spirits, beer and carbonated beverages, while it is not effective for wine. However, this general result change when looking at sub-groups of individuals. The second chapter studies how real minimum wages affect population health in China. I conclude that real minimum wages are negatively and significantly related to population health, a result which might be explained by the role of more stressful working conditions as a consequence of a higher minimum wage. Finally, the third chapter analyses the effects of incentive mechanisms on the behaviours of primary health workers and the subsequent effects on the quality of hypertension management. The empirical investigation suggests that negative financial incentives could motivate primary health workers and improve the quality of hypertension management. However, neither positive financial incentives nor different modes of performance evaluation are significantly related to the quality of hypertension management.

The master thesis focuses on the literary analysis and comparison of four books for young adults written by different authors. Two of the compared works are novels Eulengesang and Carolina: Ein knapper Lebenslauf by Iva Procházková, originally a Czech author and representative of a so called migrant literature. The thesis deals also with the novel Die Wolke by Gudrun Pausewang and the short story collection Meine freie deutsche Jugend written by Claudia Rusch. The main aim of the thesis is to compare and interpret the portayal of adolescence in the chosen works. Special attention is confined to the types of characters and situations they encounter, motivation for their behaviour and last but not least, to the motives. The opening chapters present historical overview of the Czech and German young adult literature. The thesis also includes a chapter on adolescence from the scientific perspective of developmental psychology. In the conclusion, the master thesis contrasts the chosen literary works in pairs: Eulengesang with Die Wolke and Carolina: Ein knapper Lebenslauf with Meine freie deutsche Jugend.

The present thesis focuses on the representations of gays in Czech and Slovak contemporary theater plays. Its aim is to analyze the queer and campy aspects of the representations of gays in the mainstream plays and uncover the principles of creation of the gay character in the structure of dramatic Arts. The results of analysis explain the integrative mode of representation of gays in Czech and Slovak contemporary plays.