Some diagnostic methods have been used for a very long time. Congo red stain saw the light of day in 1883, and quickly became important in many fields of use. Nowadays we recognize the importance of Congo red in diagnose of amyloid diseases. However, the technique and experience needed throughout the process from a suspected case to the diagnose is of greate importance. When diagnostic difficulties appeared in a few patient cases at the local hospital in Gävle, Sweden, a solution was needed. A delayed diagnose could have a potential devastating outcome seen in the perspective of the patient. Therefore it is crucial to have both sensitive and specific diagnostic methods that are optimized against the sought pathogenesis. This study aspired to find the solution to the difficulties in diagnostic work, brought to light by a pathology doctor at the hospital. Several different methodical procedures are used throughout the process, and were evaluated with focus lying on the thickness of the tissue, the staining method and the microscopes used in diagnostics. Different thickness of the tissue was cut and stained. The results demonstrated the importance of proper techniques and methods in preparing the tissue, and the tools to analyse it with. The thickness of tissue and the lightsource in the microscope played a cruicial role in diagnostics. Additionally it showed the importance to continue to raise the quality of work and make progress in the diagnostic and scientific field, possibly by finding new applications for old methods.

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The complement system is a part of the innate immunity. Its function is to eliminate pathogens, by proteins interacting directly with pathogen surfaces and promoting a pro-inflammatory and anti-microbial environment. Related to the lectin pathway of the complement system are ten known proteins, with component properties and disease association still unclear. The aim of this study was to evaluate the instrument WES for the detection of seven proteins associated to the lectin pathway. The novel instrument introduces an automated technique based on capillary electrophoresis and immunoblotting. Trials were performed on donor plasma using instrument associated kits. For the evaluation, these kits were combined with assorted primary and secondary antibodies from several species, as well as antibodies in biotinylated form. The high protein content of plasma caused many artefacts, affecting separation and displaying unspecific binding of both primary and secondary antibodies. Biotinylated antibodies coupled with the kit streptavidin-horseradish peroxidase showed the best results for further trials. Several issues remain to be solved in the optimization, including determining the unspecificity of biotinylated primary antibodies, best antibody concentrations and optimal sample preparation and dilution.

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In this thesis, a very flexible framework for simulating RIS is designed to beused for Infobroker testing. Infobroker is an application developed by MawellSvenska AB that connects RIS and PACS to achieve interoperability by enablingimage and journal data transmission between radiology sites. To put the project in context, the field of medical informatics, RIS and PACS systems and common protocols and standards are explored. A proof-of-concept implementation of the proposed design shows its potential and verifies that it works. The thesis concludes that a more specialized approach is preferred.

Kolistin är ett gammalt polypeptidantibiotikum och används som sista utväg för behandling av allvarliga infektioner orsakad av multiresistenta gramnegativa bakterier. Nya studier har påvisat kolistinresistensgener, mcr (mobil colistin resistance), hos extended spectrum beta-lactamase (ESBL)-producerande Enterobacteriaceae. Mcr-genen ligger i plasmider som kan överföras mellan bakterier, vilket innebär att det är mycket svårare att behandla människor och djur vid infektion orsakad av patogen som erhållit denna resistens. Syfte med detta projekt var att utveckla en PCR-baserad metod för detektion av mcr-1 och mcr-3 gener. I denna studie optimerades en PCR-metod och sedan screenades 60 ESBL-isolat från Kristianstads vattenrike. Därefter undersöktes PCR-produkt med hjälp av agarosgelselektrofores. Resultatet visade att 51oC är den optimala annealingtemperaturen vid multiplex-PCR för detektion av mcr-1 och mcr-3. Ingen av mcr-generna kunde detekteras i de 60 ESBL-isolaten. Positiva kontrollstammar med specifika primers kunde detekteras i PCR-analyser som genomfördes i denna studie, vilket tyder på att den optimerade PCR metoden kan vara pålitlig för detektion av mcr-1 och mcr-3 generna.

Cervical cancer is almost exclusively caused by the HPV virus, whit HPV 16 and 18 involved in the majority of cases. The HPV virus can be divided into high risk and low risk types, where the high risk types are most associated with cancer. HPV is spread by sexual skin to skin contact, many people get infected without getting cervical cancer. HPV is also involved in the development of several other types of cancers such as oral and other genital cancers. The HPV virus infects epithelium stem cells and disrupts basic functions of the cells. A high expression of the late genes early in an infection may result in that an HPV 16 infection dies out. The late gene expression was analysed by using a CAT ELISA method, in the cell lines used one of the late genes had been replaced by a CAT reporter gene. Several small molecules where investigated, to study the regulation of the late gene expression. The results of the study was that a regulation of the late gene expression could be seen when pBELMCAT was treated with TPA, TA and RA where TPA gave the highest increase in the late gene expression. TA/RA combined with TPA increased the expression even more. As a conclusion it seems possible for small molecules to be used in treatments for cervical cancer that is caused by HPV 16, to upregulate the late gene expression and maybe be able to eliminate the infection before serious damage and disease can develop.

Background: Overweight and obesity constitutes a threat to public health all around the world, and more individual’s dies from obesity than from hunger. This condition is increasing worldwide and obesity also occurs in children. According to WHO, there are 42 million overweight children in the world. Obesity and overweight in childhood leads to insulin resistance, elevated blood lipids and cardiovascular effects. Obesity is caused by positive energy intake, poor exercise habits, environmental factors, food habits and stress etc. It is also known that hormones influence the incidence of obesity. It has been found that individuals with overweight and obesity has elevated glucagon levels in the bloodstream Purpose: This work aims to investigate whether there is a difference in glucagon concentration between the sexes at ages 10-18 years with obesity, and if there is an association between glucagon concentration and weight, FM and FFM. Materials and Methods: In this study, obesity is determined by BIA and the caliper in combination as three compartment model. Blood samples were taken by venipuncture and glucagon were analyzed in fasting plasma. Results: BIA and the caliper together had a significant correlation with body composition. Blood glucagon was found to be high in obese children. Conclusion: Glucagon concentration in obese children correlates with weight and FM. However, further investigation is needed regard to how glucagon interact with weight, FM and FFM

Cryopreservation is used to freeze and store donor’s sperm, from men who are going throughcertain medical treatments such as chemotherapy or radiotherapy and from men withazoopermia or severe oligozoospermia. Stored sperm samples are used in artificialreproductive technologies. Formation of ice crystals is the biggest problem when freezingcells because of the risk severe damage to the cells. To guarantee optimal survival rates infrozen sperm samples, cryoprotectants which protect sperms from ice crystals formations, isadded.The main objective of the study was to compare two different freeze solutions and theirimpact on sperm survival in frozen sperm samples. In addition, an additional aim was todetermine if the amount of motile sperm changes when the samples is left for two hours inroom temperature.In this study, 31 samples were used. Each sample was divided into two groups. The firstgroup was mixed with a SpermFreeze Solution™ and the second group was mixed withSperm CryoProtectTM II. All samples were frozen using cooling vapor and stored in tankswith liquid nitrogen. The concentration of motile sperm was measured after thawing andpreparation. SpermFreeze SolutionTM showed significant better results in the number ofsperm who survived after freezing and thawing process Also, the results showed that werewas no significant difference on concentration of motile sperm in samples after being left atroom temperature for two hours. In summary, an improvement in the process of preparationrecommends to be performed to reduce the mechanical stress to ensure a greater quantity ofmotile sperm after the whole process.

The climate in the north is changing over time, which affects the nature in many ways. For instance, some microorganisms that cause infections might become more common. This might have negative consequences for reindeer husbandry. In Sweden, this is an industry that is relatively large. However, even though the reindeer is common in the north the knowledge about its diseases is limited.In this study the prevalence of microorganisms that may cause infection in reindeer was investigated. Comparisons between different sami villages and previous studies were performed to detect differences that could occur due to climate changes. The diseases and microorganisms that were analyzed with PCR were malignant catarrhal fever, herpes infections, Chlamydia sp. and bovine viral diarrhea (BVD). The cause of eye problems in reindeer was also investigated. BVD and bovine leukemia virus where analyzed with ELISA. Next generation sequencing where used for broader screening of samples for microorganisms that might be of interest of future analysis in more detailed follow-up studies.Since not enough samples were available at the time of this study findings could not be linked to changing climate. In the reindeer with eye infection Chlamydia sp., Moraxella sp. and Neisseria sp. can probably be involved causing disease. This should be further investigated to be able to determine whether it is true or not by analyzing samples from individuals without changes in the eyes. The prevalence of reindeers with antibodies against BVD has increased in Sweden since 2012. There will be further studies in this field with reindeers from other northern countries.