Extended Spectrum β-Lactamases (ESBLs) are produced by the Enterobacteriaceae bacterial family, mainly by E. coli and K. pneumoniae. As these species are some of the main causes of urinary tract infections and sepsis, ESBL-production is of major concern. Occurrence of ESBLs also gives rise to concern as it is increasing epidemically. This because the genes coding for ESBLs (i.e. bla-genes) are located on plasmids replicating and spreading the replicated copies independently. Plasmids replicate by replicons. Plasmids with the same replicon variant are grouped into the same plasmid family. The aim of this study was to detect plasmid families carrying bla-genes in E. coli and K. pneumoniae from clinical (n = 6) and environmental water (n = 22) isolates. Plasmid family prevalence was examined. Association between plasmid families and bla-genes was also examined. Plasmid families were detected by a PBRT kit (PCR Based Replicon Typing), a multiplex PCR kit that detected 30 replicons, whereof 27 replicons representing the 27 plasmid families in Enterobacteriaceae, and three novel replicons. The IncF plasmid family was the most prevalent for both species in both clinical and environmental isolates. IncF seemed to be prevalent for all examined ESBLs, but it was difficult to associate one bla-gene with one plasmid family as most isolates carried several bla-genes and several plasmid families. / Extended Spectrum β-Lactamases (ESBLs) produceras av bakteriefamiljen Enterobacteriaceae, främst av E. coli och K. pneumoniae. Eftersom dessa arter är bland de vanligaste orsakerna till urinvägsinfektioner och sepsis är ESBL-produktion ett allvarligt problem. ESBL är också oroande eftersom det sprids epidemiskt. Detta möjliggörs av att generna som kodar för ESBLs (s.k. bla-gener) ligger på plasmider, som replikerar och sprider de replikerade plasmidkopiorna självständigt. Plasmider replikeras som s.k. replikon. Plasmider med samma replikonvariant tillhör samma plasmidfamilj. Syftet med detta arbete var att detektera plasmidfamiljer som bär bla-gener i E. coli och K. pneumoniae isolerade från kliniska prov (n = 6) och miljöprov (n = 22) från Helge Å. Plasmidfamiljernas prevalens undersöktes, liksom sambandet mellan plasmidfamiljer och bla-gener. Plasmidfamiljerna detekterades med ett PBRT-kit (PCR Based Replicon Typing), ett multiplext PCR-kit som detekterade 30 replikon varav 27 replikon som representerar de 27 plasmidfamiljer som finns i Enterobacteriaceae och tre nya replikon. Plasmidfamiljen IncF var vanligast förekommande i båda arter i både kliniska isolat och miljöisolat. IncF verkade förekomma för alla undersökta typer av ESBL, men det var generellt svårt att förknippa en bla-gen med en plasmidfamilj, eftersom de flesta isolaten bar flera bla-gener och flera plasmidfamiljer.

<p>Immunoglobulin E (IgE)-mediated allergy occurs when our immune system causes a reaction to otherwise harmless substances (allergens). Allergens are predominantly proteins present in biological materials such as pollens, mites, animal epithelia, moulds and foods. </p><p><i>In vitro</i> tests for specific IgE antibodies usually employ an allergen source extract as an antibody capturing reagent. The proportion of allergenic molecules in these biochemically complex extracts may vary.</p><p>Recombinant allergens may be obtained in large quantities with biotechnological techniques. These proteins can be characterized biochemically and immunologically, resulting in tests with minimal batch-to-batch variation. This thesis describes different uses of recombinant allergens in component-resolved diagnostics (CRD).</p><p>In CRD, single allergenic proteins are used to establish a sensitization profile of the patient. Two timothy grass (<i>Phleum pratense</i>) pollen allergens, Phl p 11 and Phl p 4, were cloned and expressed as recombinant proteins. They were subsequently characterized and can, for example, be used in a panel for grass pollen CRD.</p><p>Single allergens may be useful as diagnostic markers for allergic sensitization. This phenomenon was studied using tropomyosin, a major allergen from the shrimp <i>Penaeus aztecus</i> (Pen a 1). The characteristics of the recombinant and natural proteins were compared. The recombinant tropomyosin was then extensively tested using specific competition for IgE binding against extracts of other crustacean species, house dust mite and cockroach.</p><p>In cases when an important allergen is missing or underrepresented in a natural extract, the corresponding recombinant allergen may be added to the extract as a spiking reagent. Previous studies have shown that latex extracts for diagnostic testing may lack the allergen Hev b 5. Recombinant Hev b 5 was expressed from a synthetic gene construct, incorporating several adaptations to enable efficient large scale production of the recombinant protein, to be used as a spiking reagent.</p>

Immunoglobulin E (IgE)-mediated allergy occurs when our immune system causes a reaction to otherwise harmless substances (allergens). Allergens are predominantly proteins present in biological materials such as pollens, mites, animal epithelia, moulds and foods. In vitro tests for specific IgE antibodies usually employ an allergen source extract as an antibody capturing reagent. The proportion of allergenic molecules in these biochemically complex extracts may vary. Recombinant allergens may be obtained in large quantities with biotechnological techniques. These proteins can be characterized biochemically and immunologically, resulting in tests with minimal batch-to-batch variation. This thesis describes different uses of recombinant allergens in component-resolved diagnostics (CRD). In CRD, single allergenic proteins are used to establish a sensitization profile of the patient. Two timothy grass (Phleum pratense) pollen allergens, Phl p 11 and Phl p 4, were cloned and expressed as recombinant proteins. They were subsequently characterized and can, for example, be used in a panel for grass pollen CRD. Single allergens may be useful as diagnostic markers for allergic sensitization. This phenomenon was studied using tropomyosin, a major allergen from the shrimp Penaeus aztecus (Pen a 1). The characteristics of the recombinant and natural proteins were compared. The recombinant tropomyosin was then extensively tested using specific competition for IgE binding against extracts of other crustacean species, house dust mite and cockroach. In cases when an important allergen is missing or underrepresented in a natural extract, the corresponding recombinant allergen may be added to the extract as a spiking reagent. Previous studies have shown that latex extracts for diagnostic testing may lack the allergen Hev b 5. Recombinant Hev b 5 was expressed from a synthetic gene construct, incorporating several adaptations to enable efficient large scale production of the recombinant protein, to be used as a spiking reagent.

O objetivo deste trabalho foi avaliar a potencialidade de implantação da Produção Mais Limpa (P+L), através do estudo de caso em Laboratório Biomédico de Referência em uma instituição pública de ensino e pesquisa localizada no Rio de Janeiro. Esta investigação é exploratória e analítica, utilizando-se como instrumentos a revisão bibliográfica e documental, a observação direta e a entrevista com aplicação de questionários voltados aos responsáveis pela área ambiental do laboratório pesquisado. A análise foi realizada confrontando-se os dados levantados com as recomendações da metodologia de P+L, identificando-se as lacunas e oportunidades para a melhoria dos serviços e processos de trabalho. O laboratório possui instalações modernas, organização, sistemas de avaliação da matéria-prima e insumos usados, além do gerenciamento dos resíduos. Contudo, nem todos os procedimentos são validados ou estão adequados às normas. Em geral, problemas em laboratórios dizem respeito ao uso excessivo de substâncias perigosas e ao manejo inadequado de resíduos, o qual pode ser contornado com a P+L, tendo como enfoque a prevenção da poluição e a minimização na fonte geradora. A redução do consumo de materiais e insumos, além da implantação de mudanças nos processos de trabalho, podem diminuir os custos financeiros e os impactos ambientais, como foi demonstrado no estudo. Para a melhoria da gestão dos laboratórios, recomenda-se a continuidade na aquisição, manutenção de equipamentos e infraestrutura. É importante a divulgação de informações ambientais e treinamento permanente para funcionários e alunos. A Sustentabilidade Ambiental só pode ser alcançada quando for bem entendida e absorvida por todos, sendo a alta administração das instituições a maior responsável para liderar esse processo. Para estudos futuros, propõe-se melhor definição e ampliação dos indicadores para o monitoramento e aprimoramento da gestão ambiental. Complementarmente, indicam-se estudos sobre a aquisição de conceitos pelos atores sobre a P+L e como eles podem contribuir com a Sustentabilidade Ambiental e a melhoria no ambiente de trabalho. Espera-se que esta pesquisa auxilie com o aperfeiçoamento da gestão no laboratório estudado e em instituições similares que a venham implantar a P+L. / The aim of this study was to evaluate the potential implementation of Cleaner Production (CP), through a case study in Biomedical Reference Laboratory in an education and research public institution located in Rio de Janeiro. This research is exploratory and analytical, using bibliographic and documentary tools, observation and interviews with questionnaires directed to those responsible for the laboratory researched. The analysis was performed by comparing the data collected with the recommendations of the CP methodology identifying gaps and opportunities to improve services and work processes. The laboratory has modern installations, organization, systems evaluation of raw materials and supplies, as well as waste management. However, not all procedures are validated and are suitable standards. In general, problems in laboratories relate to the excessive use of hazardous substances and the improper management of waste, which can be solved by the CP approach, with the focus on waste minimization and pollution prevention. The decrease in materials and supplies, and changes in work processes, can reduce costs and environmental impacts, as demonstrated in this study. To improve the management of these laboratories, it is recommended investments in the acquisition, maintenance of equipment and infrastructure. It is important to keep the dissemination of environmental information and promote participation of staff and students involved in the process, as well as its permanent training by offering courses focused on biosecurity work environments. The Environmental Sustainability can only be achieved when properly understood and absorbed by all, and the senior management of the institutions is most responsible for leading this process. For future studies, we propose a better definition of the indicators and their expansion in monitoring and improving environmental management. In addition, its indicate studies on the acquisition of concepts by the actors on the CP and how they can contribute to environmental sustainability and improving the work environment. Its expected that this research will assist the improvement of the management in the laboratory studied and similar institutions that will implement the CP methodology.

Trombocyter är suspenderade i plasma som innehåller antikroppar mot de blodgruppsantigen som saknas på erytrocyterna. För att minimera risken för en hemolytisk reaktion bestäms titern av anti-A och anti-B. Gelkortsteknik används för att detektera antikropp-antigensreaktioner och baseras på agglutinationer i en gel. Syftet med studien var att undersöka titern av anti-A och anti-B i trombocytenheter, samt att utvärdera en rutinanalys och utveckla en screeningmetod. I studien analyserades enheter av blodgrupp O och A. De kontrollerades mot anti-A och/eller anti-B både för IgG och IgM antikroppar. En screeningmetod utvecklades för att kunna screena O-enheterna och en gräns på 1:100 respektive 1:250 undersöktes. Resultatet kunde påvisa en stor skillnad i titer mellan O-och A-enheter. Titern skiljer sig signifikant beroende på om titern bestäms i plasma eller från den färdiga (utspädda) enheten. En screeningmetod på 1:100 påvisade att 86 % av enheterna hade bedömts som hög titer och en screeningmetod på 1:250 visade att andelen sjönk till 31 %. Geltekniken är en känslig metod och är beroende av kompetent personal vid avläsning. En del studier visar liknande resultat men andelen enheter med hög titer varierar och likaså metoder och titergräns. Detta påvisar svårigheterna i att bestämma en kritisk titer och att förutse risker hos patienten. Andra faktorer tros också kunna påverka riskerna. Införande av en screeningmetod på 1:250 kan öka antalet enheter som kan transfunderas över ABO-barriären. / Platelets are suspended in plasma containing antibodies to the blood group antigen missing on the erythrocytes. To minimize the risk of hemolytic reaction, the titrers of anti-A and anti-B are determined. The gel test is used to detect antibody-and antigen responses and is based on agglutinations in gel. The purpose was to investigate the titers of anti-A and/or anti-B in platelets. A routine analysis was evaluated and a screening method was implemented. In the study, units of blood group O and A were analyzed. They were checked against anti-A and anti-B for both IgG and IgM antibodies. A screening method was developed to screen the O-units and a limit of 1:100 and 1:250 was used. The results showed great difference in titers between O and A units. The titers differ significantly depending on whether the titers are determined in plasma or from the finished (diluted) unit. A screening method at 1:100 showed that 86 % of the units was rated as high titer while a screening method of 1:250 showed that this was reduced to 31 %. Gel technology is a sensitive method and is dependent on competent staff when reading the agglutinations. Some studies show similar results, but the proportion of high titer units, methods and critical titers varies. It proves the difficulty in determining a critical titer and predicting risks for the patient. Other factors are also believed to influence the risks. Implementation of a 1:250 screening method is believed to increase the number of units that can be transfused over the ABO barrier.

Kranskärlssjukdom (CAD) är en av de vanligast förekommande kardiovaskulära sjukdomarna och en av de dominerande dödsorsakerna hos äldre människor världen över. För att bekräfta diagnos och bedöma sjukdomens svårighetsgrad används idag flera diagnostiska strategier. Ökade hälsokostnader och lång kö för undersökningar väcker oro hos både patienter, läkare och myndigheter. Behovet av en enkel, säker och kostnadseffektiv metod som kan hjälpa till i utredning av CAD är stor. Det danska företaget Acarix utarbetade en ny apparat, CADScor®-system, som använder en icke-invasiv och strålningsfri metod för att utesluta stabil CAD baserad på akustisk fonokardiografi. Apparaten spelar in koronarblåsljud, vilket uppstår vid stenos i kranskärl, och beräknar patientens risk för CAD. Syftet med den här studien var att utvärdera metoden, testa CADScor® och beräkna apparatens sensitivitet, specificiteten samt positivt och negativt prediktivt värde (PPV och NPV). Tjugo patienter med misstänkt stabil CAD undersöktes med CADScor®-system och deras CAD-resultat jämfördes med resultatet efter myokardscintigrafi. Beräkningar visade att apparatens sensitivitet är 80 %, specificitet 60 %, PPV 40 % och NPV 90 %. Resultatet innebär att sannolikheten är 90 % för att en patient som fick CAD-score ≤ 20 är frisk. Det är tillräckligt högt för att använda CADScor® i klinisk praxis för patienter med låg risk för CAD. / Coronary artery disease (CAD) is one of the most common cardiovascular diseases and one of the dominant causes of death in older people worldwide. In order to confirm diagnosis and assess the severity of the disease, several diagnostic strategies are being used today. Increased health costs and long queues for investigations raise concerns among patients, medical doctors and authorities. A simple, safe and cost-effective method that can assist in the investigation of CAD is of major importance. The Danish company Acarix developed a new device, CADScor® system, which uses a non-invasive and radiation-free method to exclude stable CAD based on acoustic phonocardiography. The device records intracoronary murmurs, resulting from coronary stenosis, and calculates the patient's risk of CAD. The purpose of this study was to evaluate the method, test CADScor® and calculate the device's sensitivity, specificity and positive and negative predictive value (PPV and NPV). Twenty patients with suspected stable CAD were examined with CADScor® systems, and their CAD results were compared to the result after myocardial perfusion scan. Calculations showed that the device's sensitivity is 80 %, specificity 60 %, PPV 40 % and NPV 90 %. The result means that the probability is 90 % that a patient who has a CAD score ≤ 20 is healthy. It is high enough to use CADScor® in clinical practice for patients with low risk for CAD.

O objetivo deste trabalho foi avaliar a potencialidade de implantação da Produção Mais Limpa (P+L), através do estudo de caso em Laboratório Biomédico de Referência em uma instituição pública de ensino e pesquisa localizada no Rio de Janeiro. Esta investigação é exploratória e analítica, utilizando-se como instrumentos a revisão bibliográfica e documental, a observação direta e a entrevista com aplicação de questionários voltados aos responsáveis pela área ambiental do laboratório pesquisado. A análise foi realizada confrontando-se os dados levantados com as recomendações da metodologia de P+L, identificando-se as lacunas e oportunidades para a melhoria dos serviços e processos de trabalho. O laboratório possui instalações modernas, organização, sistemas de avaliação da matéria-prima e insumos usados, além do gerenciamento dos resíduos. Contudo, nem todos os procedimentos são validados ou estão adequados às normas. Em geral, problemas em laboratórios dizem respeito ao uso excessivo de substâncias perigosas e ao manejo inadequado de resíduos, o qual pode ser contornado com a P+L, tendo como enfoque a prevenção da poluição e a minimização na fonte geradora. A redução do consumo de materiais e insumos, além da implantação de mudanças nos processos de trabalho, podem diminuir os custos financeiros e os impactos ambientais, como foi demonstrado no estudo. Para a melhoria da gestão dos laboratórios, recomenda-se a continuidade na aquisição, manutenção de equipamentos e infraestrutura. É importante a divulgação de informações ambientais e treinamento permanente para funcionários e alunos. A Sustentabilidade Ambiental só pode ser alcançada quando for bem entendida e absorvida por todos, sendo a alta administração das instituições a maior responsável para liderar esse processo. Para estudos futuros, propõe-se melhor definição e ampliação dos indicadores para o monitoramento e aprimoramento da gestão ambiental. Complementarmente, indicam-se estudos sobre a aquisição de conceitos pelos atores sobre a P+L e como eles podem contribuir com a Sustentabilidade Ambiental e a melhoria no ambiente de trabalho. Espera-se que esta pesquisa auxilie com o aperfeiçoamento da gestão no laboratório estudado e em instituições similares que a venham implantar a P+L. / The aim of this study was to evaluate the potential implementation of Cleaner Production (CP), through a case study in Biomedical Reference Laboratory in an education and research public institution located in Rio de Janeiro. This research is exploratory and analytical, using bibliographic and documentary tools, observation and interviews with questionnaires directed to those responsible for the laboratory researched. The analysis was performed by comparing the data collected with the recommendations of the CP methodology identifying gaps and opportunities to improve services and work processes. The laboratory has modern installations, organization, systems evaluation of raw materials and supplies, as well as waste management. However, not all procedures are validated and are suitable standards. In general, problems in laboratories relate to the excessive use of hazardous substances and the improper management of waste, which can be solved by the CP approach, with the focus on waste minimization and pollution prevention. The decrease in materials and supplies, and changes in work processes, can reduce costs and environmental impacts, as demonstrated in this study. To improve the management of these laboratories, it is recommended investments in the acquisition, maintenance of equipment and infrastructure. It is important to keep the dissemination of environmental information and promote participation of staff and students involved in the process, as well as its permanent training by offering courses focused on biosecurity work environments. The Environmental Sustainability can only be achieved when properly understood and absorbed by all, and the senior management of the institutions is most responsible for leading this process. For future studies, we propose a better definition of the indicators and their expansion in monitoring and improving environmental management. In addition, its indicate studies on the acquisition of concepts by the actors on the CP and how they can contribute to environmental sustainability and improving the work environment. Its expected that this research will assist the improvement of the management in the laboratory studied and similar institutions that will implement the CP methodology.

Typ- 1 diabetes (T1D) är en autoimmun sjukdom med insulinbrist orsakad av nedbrytning av insulinproducerande betaceller i pankreas. Fyra olika antikroppar har identifierats som är riktade mot betacellsspecifika antigen; insulinautoantikroppar (IAA), glutamic acid decarboxylase antibodies (GADA), islet antigen2-antikroppar (IA-2A) och antikroppar riktade mot zinktransportören 8 (ZnT8A). I denna studie gjordes en jämförelse av metoderna 2screen islet cell autoantibody ELISA-kit (RSR, Cardiff, UK) och 3screen islet cell autoantibody ELISA- kit (RSR, Cardiff, UK), vars brunnar är coatade med GAD65/IA-2 antigen respektive GAD65/IA-2/ZnT8 antigen, för att undersöka ifall dessa båda kit ger jämförbar sensitivitet och specificitet i en adult population av nydebuterade patienter med T1D och friska vuxna blodgivare. RSR 2screen erhöll 1 % högre specificitet (98 %) jämfört med RSR 3screen (97 %) vid samma sensitivitet (92 %) och rekommenderas i första hand för screening av autoantikroppar i en population av vuxna patienter med ökad risk för T1D och friska vuxna blodgivare. / Type- 1 diabetes (T1D) is an autoimmune disease with insulin deficiency caused by degradation of insulin- producing betacells in pancreas. Four different autoantibodies that target beta- cell specific antigenes have been identified: insulinautoantibodies (IAA), glutamic acid decarboxylase antibodies (GADA), islet antigen2-antibodies (IA-2A) and antibodies against zinktransporter 8 (ZnT8A). In this study, a comparison between 2screen islet cell autoantibody ELISA-kit (RSR, Cardiff, UK) coated with GAD65/IA-2 and 3screen islet cell autoantibody ELISA- kit (RSR, Cardiff, UK) coated with GAD65/IA-2/ZnT8, was performed to investigate whether results from these two kits provide comparable sensitivity and specificity in an adult population of new onset patients with T1D and healthy adults. RSR 2screen obtained 1 % higher specificity (98 %) in comparison to RSR 3screen (97 %) on the same sensitivity (92 %) and is recommended primarily for screening of autoantibodies in a population of adult patients at increased risk for T1D and healthy adults blood donors.

Conflicts are part of our everyday life and the field of psychology describes how specific personality traits relate to specific conflict styles. However, the question remaining is why these relations exist? Recently, personality neuroscience has begun pinning down the neurobiology of personality traits, providing a deeper understanding of the human behavior. The present thesis utilizes the Five Factor Model (FFM; Costa &amp; McCrae, 1990) of personality to investigate the neurobiology underlying the inverse relation between the specific personality trait of Agreeableness and dominating conflict style (a conflict management style characterized by aggressiveness, authoritarianism and/or need for dominance). Agreeableness overlaps both empathy and aggression which can work as each other’s opposites in explaining conflict behaviors. The goal of the thesis was to investigate whether the inverse relation between Agreeableness and dominating conflict style can be explained by brain regions. Brain regions such as the medial prefrontal cortex and regions involving anterior cingulate appear to be the most prominent neurobiology describing the relation. Serotonin is the neural substance involved in most cortical and subcortical brain structures and it also regulates the suppression of aggression, making it an important substance both within Agreeableness and the preference for dominating conflict style. The thesis will sum up with a discussion including some limitations within the research and further aspects such the consequences of the findings will be discussed.

Syftet med studien var att jämföra hur väl transthorakal (TTE) och transesofageal ekokardiografi (TEE) kompletterar varandra vid diagnosticering av Staphylococcus Aureus (S. aureus) -endokardit (SAE). S. aureus är en av de ledande bakterierna inom vårdrelaterade infektioner. SAE blir vanligare, något forskare relaterar till att fler invasiva ingrepp görs. TTE är förstahandsalternativet för att besvara alternativt dementera frågeställningen och följs ofta av en TEE som bidrar till förbättrad bildkvalité och möjlighet att visualisera mindre strukturer. På Skaraborgs sjukhus i Skövde (SkaS) anses TEE-undersökningarna många gånger genomförs i onödan då få endokarditer hittas. Studien var jämförande med en kvantitativ ansats. Materialet har samlats in på kliniskt fysiologiskt laboratorium i Skövde. Det resulterade i elva deltagare. Båda könen inkluderades, patienter under 18 år exkluderades. Resultatet visade att fyra endokarditer diagnostiserades. Streptococcus Sanguinus var den bakterie som orsakade flest endokarditer medan S. aureus inte orsakade någon endokardit. Fem av elva patienter hade någon form av riskfaktor för endokardit. Fem av elva patienter blododlades positivt för S. aureus. Hygienen vid undersökningstillfällena, framförallt TEE är viktig då 80 % av vårdpersonalen kan vara bärare av S. aureus. TTE- och TEE-undersökningen kompletterar varandra. Kombinationen av undersökningarna gör resultatet tillförlitligt. TEE är därför av värde vid endokarditfrågeställning. / The purpose was to compare how transthoracic (TTE) and transesophageal echocardiography (TEE) complement each other in the diagnosis of Staphylococcus Aureus (S. aureus)-Endocarditis (SAE). S. aureus is one of the leading bacteria in health-related infections. SAE becomes more common, which researchers associate with more invasive interventions. TTE is a first step to confirm or rule out endocarditis and often followed by TEE-examination. At Skaraborg Hospital in Skövde (SkaS), the TEE-survey is considered unnecessary many times when not finding enough endocarditis. The study was a comparative study with quantitative approach, where the material was collected at clinical physiological laboratory at SkaS, Skövde, which resulted in eleven participants. Both sexes were included, patients below 18 years were excluded. Results showed that four endocarditis was diagnosed. Streptococcus Sanguinus was the bacterium that caused most endocarditis while S. aureus did not cause endocarditis. Five of eleven patients had some form of risk factor for endocarditis. Five of eleven patients were positive for S. aureus. Hygiene at the examinations, especially TEE is important because 80 % of healthcare professionals could be carriers of S. aureus. The TTE and TEE survey complement each other. Carrying out the surveys together is what makes the diagnosis safe.