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Humana trombocytantigen (Human Platelet Antigens, HPA) är genetiska polymorfismer som uttrycks på trombocyters membranglykoproteiner. Om icke-kompatibla trombocytantigen införs i blodcirkulationen kan alloimmunisering uppstå där antikroppar produceras mot främmande antigen, det kan därför vara av betydelse att registrera blodgivares antigen. Studiens syfte var att utvärdera prevalensen av HPA1a-negativa blodgivare i Jönköpings- och Östergötlands län med hjälp av flödescytometrisk fenotypning. Studien inkluderade totalt 300 blodgivare, varav 150 från Jönköpings län och 150 från Östergötlands län. Fluorokrommärkta antikroppar riktade mot CD42a och CD61 användes för att detektera HPA1a-negativitet med flödescytometrisk metod. I Jönköpings län detekterades fyra (2,7 %) HPA1a-negativa blodgivare och i Östergötlands län detekterades sex (4,0 %) HPA1a-negativa blodgivare. Statistisk analys visade ingen signifikant skillnad mellan antal HPA1a-negativa blodgivare i de undersökta länen. Länens prevalens av HPA1a-negativitet motsvarade den genomsnittliga prevalensen i Sverige och nya HPA1a-negativa blodgivare har registrerats vilket är en viktig tillgång då patienter är i behov av trombocytkoncentrat. Då studien begränsades av dess ringa storlek samt att kvinnor exkluderats bör vidare studier med större populationer och fler län i Sverige utföras.

Transferrin är ett protein som har uppgiften att transportera järn i kroppen. Transferrin är ett glykoprotein med två kolhydratkedjor med två, tre förgreningar med terminala sialinsyror. Antalet sialinsyror ger proteinet olika isoformer och benämns därefter, så som tetrasialotransferrin som har fyra terminala sialinsyror och är den vanligaste förekommande isoformen. Disialotransferrin används som en biokemisk markör för att identifiera individer som har en hög alkoholkonsumtion under en längre period. Disialotransferrin ökar i koncentrationen vid hög alkoholkonsumtion under minst 14 dagar. Den benämns som kolhydratfattigt transferrin (eng. Carbohydrate-Deficient Transferrin, CDT) och analyseras med jonbyteskromatografi. Referensvärdet för CDT är > 2,0 % då en individ har ett alkoholmissburk. Syftet med studien var att undersöka om svarstiden går att förkorta genom att optimera upparbetningen av prover och sedan utföra en metodverifiering. Optimeringen av upparbetningen omfattade en kortare inkuberingstid från nästan ett dygn till 60 minuter, förenklad tillsättning av fällningsreagens och förändrad centrifugering av proverna. I ett inledande försök analyserades 25 serumprover med olika varianter av optimerad upparbetning och resultatet jämfördes mot standardmetoden. Metodverifieringen bestod av analys av 35 serumprover med den nya upparbetningsmetoden vilket jämfördes mot standardmetoden. I analysen jämfördes även precisionen för låg (1,4 % CDT) och hög (3,2 % CDT) kontroll samt två patientprover med ett låg och hög halt av disialotransferrin med den nya metoden. Precisionen för den låga kontrollen och patientprov 1 (CV=18,05 %) visade sig var sämre än för den höga kontrollen och det patientprov 2 (CV=5,79 %). Analys av de 35 proverna visade att det var en bra överensstämmelse mellan metoderna; korrelationskoefficienten var 0,997 och ett parat t-test kunde inte detektera någon statistik signifikant skillnad mellan proverna (95 % konfidensnivå). På grund av den sämre precisionen för låga koncentrationer av disialotransferrin behöver dock den nya upparbetningsmetoden utvärderas ytterligare. Däremot är det bestämningen av CDT-halten runt 2 % som är viktig och den nya metoden har inte sämre variationskoefficient (CV %) än standardmetoden.

Zebrafish are becoming increasingly popular to use in different kinds of research projects as research animals, replacing rodents in many fields. When using animals for research, it is important to keep track of the animal health in order to get reliable results. The purpose of the study was to investigate whether these pathogens could be found analyzing environmental materials with real-time PCR instead of euthanizing fish and submitting them for histopathology. Also, to see if any material differentiated from the rest regarding accessibility to work with in a routine diagnostic laboratory.     This study was performed on environmental samples, such as filters, swabs, detritus and water, from a recirculating water system holding zebrafish. The pathogens analyzed were Mycobacterium chelonae, M. haemophilum, M. abscessus, M. marinum, M. fortuitum and Pseudoloma neurophilia, all common pathogens that can affect zebrafish.     All materials tested gave at least one positive result for most of the pathogens tested. Two pathogens were not detected, M. marinum and M. abscessus. Due to poorly working PCR-system for M. fortuitum, the results for that bacteria were deemed inconclusive. The filter materials and the swabs of the filter materials gave the best results in this small study, although all materials gave satisfactory results. In conclusion this study shows that environmental samples can be used to detect pathogens in zebrafish, but larger studies should be performed to better evaluate which material is the best one to use.

Respiratory diseases in cattle can cause economic losses due to the decreased dairy and meat production. Virus is the main reason for these diseases. Symptoms can be fever, cough and nasal discharge.     Influenza are a group of viruses belonged in the Ortomyxoviridae family. The big influenza groups are influenza A, B and C. The viruses can cause respiratory signs, and mammals can be affected. Recently a new influenza virus was found in the United States. The influenza virus was found in swine, but the natural host was later considered to be cattle. The virus was named influenza D. Different studies worldwide have confirmed the virus in a variety of regions. Antibodies have also been reported.     In this study, virologic and serologic methods were used to detect if influenza D circulates among cattle in Sweden. The serologic method performed was indirect ELISA. Serum and milk samples were investigated in the ELISA method. For the virologic detection a real-time RT-PCR was made, with a variety of study material.     Antibodies against influenza D were found in both serum and milk samples. No virus was found in the real-time RT-PCR. In Sweden the animal keeping is different compared to several other nations. For instance, the conditions of health and hygiene are better in Sweden, this may be an important cause of a system more resistant against spreading of infections. Influenza D could be more common in Sweden, but in that case further researches are needed to determine the prevalence.

Background: Acute spinal trauma is followed by a secondary injury that causes additional damage to the tissue. The mouse neuronal hybrid cell line NSC-34 is planned for studies regarding this process, wherefore the cell line needed to be established in the laboratory and a proof-of-concept study needed to be performed. A suitable target gene for this study was Neuronal Nucleus (NeuN), a neuronal marker expressed in nearly all neuronal cells although not yet studied in NSC-34. Aim: The aim of this project was to characterize the expression of NeuN in differentiated and undifferentiated NSC-34 cells and silence gene expression by using siRNA. Methods: RT-qPCR was used to measure NeuN expression during passages 5 to 15 and a comparison was performed between one early and one late passage. Lipofectamine® RNAiMAX was used for siRNA-treatment in different concentrations and several different medium compositions were tested as differentiation media. Results: NeuN was expressed in passages 5 to 15, with decreased expression levels in passage 13 (ΔCt 15.36 ± 0.16) compared to passage 5 (ΔCt 15.09 ± 0.16), p < 0.05. The expression levels did not change after differentiation. siRNA-treatment yielded knockdown when using  high concentrations of the reagent (p < 0.05). Conclusion: NeuN was expressed in a stable, low level throughout passages 5 to 15 with a slightly decreased expression during later passages and no change after differentiation. The siRNA-treatment suppressed gene expression, although further optimization is needed to increase the suppression.

Abstract Introduction This study evaluated the effect of the polymer polyvinylalcohol-carbazate (PVAC) on the bacteria Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa. PVAC is a polymer with a carbazate moiety that neutralizes free aldehydes and has shown great promise in stabilizing erythrocytes during long term storage. It has also been shown to reduce intraperitoneal adhesions after trauma. For this study, two Gram positive and two Gram negative bacteria strains were used with PVAC to evaluate its effect. Materials and methods PVAC was obtained from the research team at Ångström Laboratory, Uppsala. The bacteria were obtained from Clinical Microbiology and Hospital Hygiene, Academic Hospital, Uppsala. The methods used were spectrophotometric assessment of bacteria growth, use of FITC-conjugated PVAC to study adherence to bacteria, use of FITC-antibodies to study PVAC’s effect on bacterial adherence to erythrocytes and a qPCR for quantification of E. coli. Results and discussion PVAC displayed a clear effect of inhibition of bacteria growth in the study as shown by use of spectrophotometric assessment. Trials with FITC-PVAC showed that the polymer adheres directly to the bacteria, displaying a possible function of its inhibitory properties. The qPCR assay was able to detect the bacteria in all the dilutions used. Introduction This study evaluated the effect of the polymer polyvinylalcohol-carbazate (PVAC) on the bacteria Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa. PVAC is a polymer with a carbazate moiety that neutralizes free aldehydes and has shown great promise in stabilizing erythrocytes during long term storage. It has also been shown to reduce intraperitoneal adhesions after trauma. For this study, two Gram positive and two Gram negative bacteria strains were used with PVAC to evaluate its effect. Materials and methods PVAC was obtained from the research team at Ångström Laboratory, Uppsala. The bacteria were obtained from Clinical Microbiology and Hospital Hygiene, Academic Hospital, Uppsala. The methods used were spectrophotometric assessment of bacteria growth, use of FITC-conjugated PVAC to study adherence to bacteria, use of FITC-antibodies to study PVAC’s effect on bacterial adherence to erythrocytes and a qPCR for quantification of E. coli. Results and discussion PVAC displayed a clear effect of inhibition of bacteria growth in the study as shown by use of spectrophotometric assessment. Trials with FITC-PVAC showed that the polymer adheres directly to the bacteria, displaying a possible function of its inhibitory properties. The qPCR assay was able to detect the bacteria in all the dilutions used.

Enterohemorrhagic E. coli, EHEC, is a verotoxin producing, zoonotic pathogen, which causes diseases in humans such as bloody or watery diarrhea. Microorganisms compete for limited living space, nutrients and other resources and therefore other microorganisms are EHECs biggest competitors. To avoid outbreaks and infections with EHEC, one possible approach is to use harmless but competitive bacteria as probiotics. Therefore, the aim of this study was to evaluate three probiotic E. coli strains and their ability to outcompete EHEC in bovine feces.     Ten different cattle fecal samples from three different farms were used to mix with the three probiotic and EHEC strains. The mixture was diluted and cultivated at 0 h as a control and then incubated for 48 h at 20°C and 37°C before dilution and cultivation on CT-SMaC. Colonies was counted and ratios between EHEC and probiotic E. coli before and after incubation were calculated. Kruskal-Wallis test with Dunn’s test as post hoc test were used to see if observed reductions of EHEC were significant or not.     In 37°C, strain 10 was the only strain producing a significant reduction of EHEC. In contrast, no significant reduction was observed at 20°C in any of the strains.     Future research studying other factors and performed on live cattle models are necessary to confirm the usefulness of the studied probiotic candidates. However, these results indicate probiotics can be a useful tool to avoid infections and big outbreaks of EHEC in the future.