Förhöjda halter av endotoxin i blodet förekommer ofta vid sjukdomen pyometra hos tik och kan orsaka många av de allmänna symtom som ses vid sjukdomen. Endotoxin Activity Assay (EAA) är en ny, snabb metod för mätning av endotoxinaktivitet i helblod. EAA-metoden är anpassad för humanpatienter, men har testats i forskningsstudier och visats fungera även för hundar och efter viss modifiering också för hästar. Två olika apparaturer från finns att tillgå för EAA (från Spectral Diagnostics Incorporated). Den apparatur som utvärderas i den här studien har aldrig tidigare testats på djurslaget hund.

The problem of finding targeted medicine is a central problem in chemotherapy. From this point of view the ubiquitin-proteasome system is a highly promising object in the pharmaceutical approach. Proteasome plays a critical role in cellular protein degradation, cell cycle and apoptosis regulation.Proteasome inhibitors are substances blocking the actions of proteasome. Cancer cells are more sensitive to inhibition of the ubiquitin-proteasome system than normal cells. Therefore proteasome inhibitors have the potential to be successfully used in the cancer treatment.The study aimed to test various substances to identify possible proteasome inhibitors with the IncuCyteTM FLR image system and fluorometric microculture cytotoxicity assay.

Serum thymidine kinase 1 (TK1) activity is used as a tumor marker in disease monitoring in veterinary and human medicine. TK1, an intracellular enzyme, is involved in a salvage pathway of DNA precursor synthesis. TK1 is used in DNA precursor production by catalyzing the transfer of the gamma-phosphate-group from a phosphate-donor to the 5?- hydroxyl-group of thymidine forming thymidine-monophosphate. Nucleoside monophosphosphates are finally converted into thymidine-triphosphates. TK1 activity significantly rises in the G1 and the S phase of the cell cycle.

Glutathione (GSH) is a molecule that consists of three amino acids: glutamic acid, cysteine and glycine. GSH has several important functions: to protect cells from free radicals, reactive oxygen species and oxidative stress. GSH exists in a reduced form, GSH, and in an oxidized dimeric form, glutationdisulfid, GSSG. The enzymes glutathionereductase (GR) catalyses the reduction of GSSG back to GSH. Nicotinamide adenine dinucleotide phosphate (NADPH) is required as a coenzyme in the reaction.

Oxochlorates are anions with a partially naturally occurrence in nature but are also spread by human activities, including the paper industry. These compounds are harmful to both nature and humans, which makes it necessary to find a good way for their degradation. There are two different kinds of bacteria that can use oxochlorates as electron acceptors in their metabolism, bacteria that break down perchlorate and bacteria that break down both perchlorate and chlorate. A bacterium that can break down chlorate under anaerobic conditions is Ideonella dechloratans which holds the genes for chlorite dismutase and chlorate reductase which are enzymes for the degradation of chlorate. Gene expression and enzyme activity of chlorite dismutase are induced under anaerobic conditions, which makes it interesting to find out how this regulation functions in order to better exploit these bacteria in biological wastewater treatment.

Background: As a part of a national health control program, Statens VeterinärmedicinskaAnstalt performs diagnostics to screen flocks for certain pathogens causing high mortality,morbidity and/or serious economical losses. There are several viruses in the programincluding IBDV (infectious bursal disease virus), IBV (infectious bronchitis virus) and NDV(Newcastle disease virus). Method: 96 serum samples were collected from different poultryflocks in Sweden and analyzed by ELISA, which are currently used in the health controlprogram as well as by a commercial prototype of a multiplex immunoassay manufactured byLuminex Corp., which is currently under evaluation at the United States Department ofAgriculture USDA. This 4-plex assay detects antibodies for the three above-mentionedviruses as well as antibodies of avian reovirus. In the context of this study the ELISAs run inroutine diagnostics as well as a REO ELISA were used as the standard for comparison.Result: The antibody concentration in serum from vaccinated chickens was high while theantibody concentration level in serum from not vaccinated chickens was low.

Influenza is a viral infection that affects global health and economy with its endemic and sometimes pandemic spread. Rapid detection of Influenza viruses enables antiviral use and can bring financial savings. It is also essential for the global surveillance of prevalent Influenza strains. RT-PCR is considered the most specific and sensitive method for detection of Influenza, but Influenza mutates at a high rate and it is therefore crucial that RT-PCR methods are updated regularly.In 2014, Cepheid released their Xpert Flu/RSV XC assay, which can detect Influenza A and B and RSV by multiplex RT-PCR in approximately one hour. The aim of this study was to evaluate this assay at Laboratoriemedicin Västernorrland by using the laboratory?s previous PCR assay for detection of Influenza viruses as reference method.Real-time RT-PCR was used to compare Xpert Flu/RSV XC to the reference method.

The ability to detect protein-based biomarkers, which are linked to different diseases like colorectal cancer, is very important as a diagnostic tool. Usually complex biological samples like blood are studied which will contribute to different technical issues when performing an assay. The aim with the project is to optimize and develop the high throughput protein detection technique Proximity Extension Assay, PEA, into a 96-plex panel, in hopes of discovering an expression profile for colorectal cancer. PEA was developed by Olink Bioscience and allows specific proteins in a sample to be quantitatively transformed into nucleic acid sequences that are subsequently detected and quantified with real-time PCR. Two proximity probes containing oligonucleotide sequences bind pairwise to target protein and when brought in proximity, a DNA polymerase will extend a hybridization arm from one probe over to the second forming a double-stranded DNA sequence that can serve as a template in real-time PCR.

Food-poisoning is a major health problem and an estimated half a million Swedes are food-poisoned annually, with acute gastroenteritis as a consequence. One of the major causes of contaminated foods is related to food- and waterborne viruses. To be able to trace back the source of contaminant, the method of detecting viruses must be specific and sensitive. No standardized method for detecting foods for sapovirus exists today. The aim of the work described in this bachelor thesis is to implement and opti-mize a real-time RT-PCR method for the detection of all genogroups of human sapovirus in foods.

Background: As a part of a national health control program, Statens VeterinärmedicinskaAnstalt performs diagnostics to screen flocks for certain pathogens causing high mortality,morbidity and/or serious economical losses. There are several viruses in the programincluding IBDV (infectious bursal disease virus), IBV (infectious bronchitis virus) and NDV(Newcastle disease virus). Method: 96 serum samples were collected from different poultryflocks in Sweden and analyzed by ELISA, which are currently used in the health controlprogram as well as by a commercial prototype of a multiplex immunoassay manufactured byLuminex Corp., which is currently under evaluation at the United States Department ofAgriculture USDA. This 4-plex assay detects antibodies for the three above-mentionedviruses as well as antibodies of avian reovirus. In the context of this study the ELISAs run inroutine diagnostics as well as a REO ELISA were used as the standard for comparison.Result: The antibody concentration in serum from vaccinated chickens was high while theantibody concentration level in serum from not vaccinated chickens was low.

Background: As a part of a national health control program, Statens VeterinärmedicinskaAnstalt performs diagnostics to screen flocks for certain pathogens causing high mortality,morbidity and/or serious economical losses. There are several viruses in the programincluding IBDV (infectious bursal disease virus), IBV (infectious bronchitis virus) and NDV(Newcastle disease virus). Method: 96 serum samples were collected from different poultryflocks in Sweden and analyzed by ELISA, which are currently used in the health controlprogram as well as by a commercial prototype of a multiplex immunoassay manufactured byLuminex Corp., which is currently under evaluation at the United States Department ofAgriculture USDA. This 4-plex assay detects antibodies for the three above-mentionedviruses as well as antibodies of avian reovirus. In the context of this study the ELISAs run inroutine diagnostics as well as a REO ELISA were used as the standard for comparison.Result: The antibody concentration in serum from vaccinated chickens was high while theantibody concentration level in serum from not vaccinated chickens was low.

Abstract Tripeptidyl-peptidase II (TPPII), is present in most eukaryotic cells. It cuts tripeptides from the N-terminus of peptides and is especially important for degrading peptides longer than 15 amino acids. TPPII also tailors long peptides into suitable substrates for the enzymes which transport and produce the peptides that MHC I present. Increased levels of TPPII have also been found in certain cancer cells, thus it is of interest to determine if TPPII could be used as a tumour marker.The aim of this study was to optimise and evaluate 3 different methods for quantifying TPPII. Western blot, enzyme-linked immunosorbent assay (ELISA) and fluorophore-linked immunosorbent assay (FLISA) protocols were optimised regarding incubation times and antibody dilutions.

Background: Hepatitis E (HEV) is a small, non-enveloped virus that belongs to the viral genus Hepevirus. HEV is a positive sense single-stranded RNA virus and there is insufficient information regarding its replication. This is mainly because the virus has low capacity to grow in normally used cell cultures. Many specific strand assay detection studies have been done in order to understand more about HEV replication. Unfortunately, these assays have the disadvantage of giving false positive results.Aim: The aim of this project was to improve the positive strand assay to increase specificity and eliminate false positivity which is due to high sensitivity of the polymerase chain reaction (PCR).

A quart of the of the adult population worldwide havemetabolic syndrome. A sedentary lifestyle, low physicalactivity, combined with wrong eating habits, stress,smoking and psychosocial factors, are the main causes ofdevelopment of the metabolic syndrome. The purpose ofthe literature review was to illuminate the physicalactivity in persons who have the metabolic syndrome, andwhat sort of advice a nurse use in the promotion ofphysical activity. The study was conducted as a literaturereview, including 15 scientific quantitative articles. In tenarticles, the focus was on physical activity at themetabolic syndrome, and five articles focused on variousforms of counselling for physical activity.

Objective:This study aimed to investigate the correlation between physical activity level (PAL), measured with activity diary and accelerometer, among overweight and obese children and adolescents. The aim was also to study differences in PAL between girls and boys.Design:55 children and adolescents, whom been subjected to a physical activity registration with a physical activity diary and accelerometer during three or four days, were included in this study. From the data received from the children?s journal records PAL was calculated and compiled for statistic analysis.Results:The results of the study showed that the children underestimated their physical activity level when measured with activity diary, in comparison with the physical activity level measured with accelerometer. Differences between girl and boys PAL values were not statistically significant, although the girls PAL values from the accelerometer, but not from the activity diary, reached a moderate activity level according to Nordic nutrition recommendation.