Fästingburna infektioner är ett ökande problem, och därmed även infektioner orsakade av Rickettsia spp. Syftet med studien var att undersöka risken att drabbas av en infektion efter bett av rickettsiainfekterad fästing. Specificitet och sensitivitet av Fluidigm-PCR jämfördes mot real time polymerase chain reaction (realtids-PCR) vid detektion av Rickettsia spp. i fästingar som bitit människor. Vidare undersöktes om det finns någon korrelation mellan fästingens blodsugningstid och serokonversion mot Rickettsia spp. 753 fästingar lämnades in av 104 deltagare i Sverige och på Åland. Fästingarna analyserades med realtids-PCR för att detektera gltA-genen som är specifik för Rickettsia spp. 3,5 % av fästingarna var positiva för Rickettsia spp. med realtids-PCR. Vid analysering med Fluidigm-PCR av samma material blev 1,3 % av proverna positiva. Beräkningar som gjordes med realtids-PCR som referens visade att Fluidigm-PCR har sämre specificitet och sensitivitet jämfört med realtids-PCR. Deltagare som serokonverterade (n=5) lämnade endast in rickettsianegativa fästingar som därför inte kunde kopplas till infektionen. Därmed kunde inga slutsatser dras om risken att drabbas av en infektion efter bett av rickettsiainfekterad fästing eller om det föreligger någon korrelation mellan fästingens blodsugningstid och serokonversion. / Tick-borne infections are increasing, including infections caused by Rickettsia spp. The aim of this study was to examine the risk of developing an infection after being bitten by a tick infected with Rickettsia spp. Specificity and sensitivity of a Fluidigm-PCR assay were compared to real time polymerase chain reaction (real-time PCR) assay when detecting Rickettsia spp. in ticks that had bitten humans. Possible correlation between the tick's feeding time and seroconversion against Rickettsia spp. was also investigated. A total of 753 ticks from 104 participants in Sweden and the Åland Islands (Finland) were analyzed with real-time PCR to detect the gltA gene specific for Rickettsia spp. 3.5 % of the samples were positive for Rickettsia spp. with real-time PCR, while only 1.3 % of the samples were positive with Fluidigm-PCR. Calculations showed that Fluidigm-PCR assay has lower specificity and sensitivity than the real-time PCR assay. Unfortunately, no conclusions could be drawn considering correlation between the tick's feeding time and seroconversion of the bitten humans since no participants who had seroconverted had also submitted ticks containing Rickettsia spp. Therefore, no conclusions could be drawn considering the risk of developing an infection after being bitten by a tick infected with Rickettsia spp. / STING-studien

Humanpatogena stammar av bakterien Yersinia enterocolitica kan orsaka akut gastroenterit. För identifiering av bakterien odlas fecesprover ut på CIN-agar. På senare år har en kromogen agarplatta framtagits som differentierar mellan patogena och apatogena stammar av Y. enterocolitica. Syftet med studien är att jämföra och utvärdera två CIN-agar, med agarbaser och supplement från två olika företag (Liofilchem och Oxoid), och CHROMagar Y. enterocolitica (CHROMagar). Odling av fecesprover samt seriespädning av sex Y. enterocolitica stammar och en Y. pseudotuberculosis utfördes. Vid utodlade fecesprover jämfördes växt och hämning av övriga bakterier. Vid seriespädning räknades antal kolonier på plattorna för respektive spädning, samt utseende av kolonier på plattor bedömdes. Resultatet tyder på att skillnad av hämningseffekt av Y. enterocolitica och utseende på kolonierna finns mellan de två CIN-agarplattorna. Oxoid’s CIN-agar erhöll större kolonier, lägre hämningseffekt av Y. enterocolitica och detektionsgräns än Liofilchem’s CIN-agar. På CHROMagar-plattan växte de patogena stammarna med bleklila kolonier och de apatogena stammarna med blåa kolonier. Hämningseffekt av Y. enterocolitica hos CHROMagar-plattan är densamma som Oxoid’s CIN-agar. Slutsatsen är således att Oxoid’s CIN-agar och CHROMagar har samma hämningseffekt av Y. enterocolitica men CHROMagar differentierar mellan patogena och apatogena stammar. Liofilchem’s CIN-agar har högre hämningseffekt än CHROMagar och Oxoid’s CIN-agar. / Pathogenic strains of Yersinia enterocolitica can cause acute gastroenteritis in humans. To identify the bacterium, cultivation of stool samples on CIN-agar are performed. A chromogenic medium has been developed that differentiate between pathogenic and nonpathogenic strains of Y. enterocolitica. The purpose is to compare and evaluate two CIN-agar, with agar bases and supplements from two companies (Liofilchem and Oxoid), and CHROMagar Y. enterocolitica (CHROMagar). Growth of stool samples and serial dilutions of six Y. enterocolitica strains and one strain of Y. pseudotuberculosis were performed. Comparisons of the growth and inhibition of other bacteria were done for the stool samples. Colonies for each dilution were counted and appearance of the colonies was evaluated. The result indicates that a difference in inhibitory effect on Y. enterocolitica and appearance of colonies exist between the two CIN-agar. All strains grew with larger colonies on Oxoid CIN-agar than on Liofilchem’s. Oxoid CIN-agar and CHROMagar have a lower inhibitory effect on Y. enterocolitica than Liofilchem’s. On CHROMagar, the pathogenic strains grew with mauve colonies, whilst the nonpathogenic strains grew with blue colonies. Thus, the conclusion is that CHROMagar and Oxoid CIN-agar have less inhibitory effect on Y. enterocolitica than Liofilchem’s. CHROMagar can differentiate between pathogenic and nonpathogenic strains.

HPA1a är ett antigen på trombocytytan som kan orsaka alloimmunisering, såsom neonatal alloimmun trombocytopeni och post-transfusion purpura, vilket kan ge svåra blödningssymptom. I fall med antikroppar mot HPA1a måste kompatibla trombocyter finnas tillgängliga. Syftet med studien var att etablera en flödescytometrisk screeningmetod för fenotypning av HPA1a antigen på trombocyter samt att finna HPA1a negativa blodgivare. Före flödescytometrisk analys poolades två till fem blodprover samman till ett prov och vid förekomst av HPA1a negativa trombocyter i poolen analyserades proverna individuellt. Fluorokrommärkta anti-humana antikroppar mot CD42a och CD61 användes för att särskilja HPA1a negativa trombocyter från HPA1a positiva. Totalt fenotypades 177 blodprover varav sju (4%) typades som HPA1a negativa. Av de sju fynden genotypades fyra vid ett externt laboratorium vilket bekräftade att de var HPA1a negativa. Flödescytometrisk screening av HPA1a är snabb, pålitlig och lämplig för storskalig screening. För att fastställa prevalensen av HPA1a negativa individer behöver mer omfattande studier utföras där en större population ingår. Att ha flera HPA1a negativa blodgivare registrerade ger möjligheten att hjälpa patienter i andra regioner. / HPA1a is an antigen on the platelet surface that can cause alloimmunization, such as neonatal alloimmune thrombocytopenia and post transfusion purpura, which can cause severe bleeding symptoms. In case of antibodies against HPA1a, compatible platelets must be available. The purpose of the study was to establish a flow cytometric screening method for phenotyping HPA1a antigen on platelets and to find HPA1a negative donors. Before the flow cytometric analysis, two to five blood samples were pooled into one sample and in the presence of HPA1a negative platelets in the pool, the samples were analyzed individually. Fluorochrome –labeled anti-human antibodies to CD42a and CD61 were used to distinguish HPA1a negative platelets from HPA1a positive. A total of 177 blood samples were phenotyped, of which 7 (4%) were HPA1a negative. Of the seven findings, four samples were genotyped at an external laboratory confirming that they were HPA1a negative. Flow cytometric screening of HPA1a is fast, reliable and suitable for large scale screening. In order to determine the prevalence of HPA1a negative individuals, more extensive studies need to be performed involving a larger population. By having many registered HPA1a negative donors, it can provide opportunities to help patients in other regions.

Background: Overweight and obesity is increasing worldwide and more and more peopleare dying due to their overweight rather than of starvation. Obesity in young children isbecoming more common and the physical activity is decreasing with age. To measurechildren’s daily physical activity sveral methods can be used, such as Actical activity monitorand activity diary.Purpose: The aim of this study was to investigate if there is a difference between the twomethods mentioned above, current physical activity, age and gender in obese children and toinvestigate how many of these children that reached the recommendations of daily activity.Materials and Methods: All included were subjected to the two activity methods,collection of data from anthropometric measurements and body composition measurements.In total 122 children between the ages 10 to 17 years, 38 girls and 67 boys where included inthis study.Results: None of the children reached the recommended daily activity. Correlations betweenobjective and subjective methods showed a stronger association with the activity monitor thanwith the activity diary. The study showed that girls had a higher fat mass (FM) than boys,while boys had higher fat free mass (FFM).Conclusion: This study showed that physical activity decline with age and boys had higherdaily physical activity. Boys had more tendencies to improve their fat free mass. Girls hadhigher fat mass and lower fat free mass. Which method that gives the most adequate resultsrequires a larger population and more studies of the topic.

Transfusion of platelet concentrates (PC) can be necessary for patients to maintain coagulability. It is vital that the platelets maintain viability and function during processing and storage to obtain enhanced coagulability in the transfused patient. Today, no test is used to verify platelet function in either donors or in PC’s. Observing swirling effect is the only test applied to control platelets before transfusion but the method is based on platelet morphology and does not directly evaluate platelet function.Impedance aggregometry (IA) (Multiplate analyzer, Roche Diagnostic) is a promising method for measuring platelet function, measuring changes in impedance over time when platelets adhere to electrodes. IA has been well evaluated for the purpose of analyzing whole blood but analyzing PC’s is a relatively new application of the method.Samples from platelet donors and PC’s were analyzed with IA to evaluate correlation in function between donors and PC’s, in the hope of being able to predict function in PC. Different platelet concentrations were also analyzed to evaluate the impact of varying concentration on impedance. Adenosine diphosphate (ADP), collagen and thrombin receptor activating peptide 6 (TRAP-6) were used to induce aggregation. Platelet function was measured in PC’s on day 1 and 4 after donation.A significant correlation was observed between platelet function in donors and in PCs on day 1, measured with ADP. An important finding was also that platelet concentration does affect impedance, in collagen-induced aggregation more than ADP-induced. It is therefore possible that a correlation would also have been found between donors and PC’s analyzed with collagen if the platelet concentration would have been standardized.

In today’s medical science studies, many tumor markers are being used to monitor cancer cell proliferation, but the number of assays for analysis of these markers are few. The aim of this study was to find an easier and more time-efficient way to measure the activity of a specific tumor marker called tymidine kinase 1 (TK1). This tumor marker is an important enzyme involved in cell proliferation and is a key enzyme in the salvage pathway. TK1 activity is related to the occurrence of hematological malignancies and cell activity and therefore have been used as a marker when monitoring this group of patients in treatment. Measurement of the enzyme activity in this study was performed by using an oligonucleotide assay. Detection of the enzyme activity in whole blood and in plasma has not previously been shown. The TK1 activity measured in whole blood and plasma correlated with TK1 activity measured in serum (R2=0,8651 and R2 =0,9845, respectively). It was found that it is possible to determine the TK1 activity in whole blood but only if the activity was measured on the same day as the blood samples were taken. The results shows that the activity measurement of TK1 in plasma and whole blood can be used as a marker to verify patients' therapy in cancer care. This study is only the beginning and further investigations should be made in the future to determine if the method that is subject to this study has the requested effects.

The routines for blood testing were changed during 2010 at the blood bank in UAS. At first, the blood test was taken before the donation and now it is taken after donation. Along with this, the blood bank increased the lowest level for allowance of blood donation with 10 g/L both for men and women. The level is now on 125 g/L and 135 g/L for women respectively men. After the increase, it was noticed that the amount of blood donors were deferred due to low Hb levels in creased. A study made in year 2013, investigated how much the Hb-levels actually was changed during a blood donation. It showed that it was lowered in means by 6 g/L and not 10 g/L as previously thought.The aim of this study was to see if the sitting position of the blood donor during waiting time and the supine position during the time of blood donation may had any effect on the difference of the Hb-level during the blood donation.Data from the 120 blood donors in the earlier study was collected. Hb values, before and after blood donation, were taken from the earlier study and registered times were taken from the database Prosang. The waiting time, time of blood donation and the difference of Hb-levels were calculated and correlated with Spearmanns correlation coefficient.The results did not show any correlation between the times and the difference in Hb-levels. One of the reasons may be that the blood donor physiology differ and thus the change in Hb-level can vary.

The aim of the current study was divided into two separate goals, (i) optimization of a number of PCR-based protocols employed for genotyping transgenic mouse lines and (ii) evaluating a protocol for co-detection of mRNA and its correlated protein in the mouse midbrain. The optimization was performed on PCR protocols for genotyping the following transgenic mouse lines; Dat-Cre, Vglut2-Lox, Vglut2-Cre and Vmat2-Lox. Also, two different polymerases were evaluated parallel to each other – KAPA and Maxima Hot Start. One of the main findings from the PCR optimizations were that for the Vglut2-Lox protocol. By decreasing the annealing temp and increasing the MgCl2 the bands appeared brighter.  For the second part of the project, in-situ hybridization (ISH) was used to detect the mRNA expression with a `non-radioactive in situ hybridization´ protocol, using digoxigenin or fluorescein labelled riboprobes (mRNA probes). To detect the correlated protein a basic immunohistochemistry (IHC) protocol with the use of primary and secondary antibodies was implemented. The combined protocol was tested with Nd6 and Grp markers. Before testing to combined the protocols the ISH protocol was performed alone with riboprobes for Girk2, Lpl and Fst. The combined protocol detected mRNA and protein for both the control marker Th and the Nd6 marker. In conclusions, the optimized PCR protocols were optimal when used with the Maxima Hot Start polymerase and the new combined ISH and IHC protocol worked for markers Th and Nd6.

I denna studie jämfördes två olika metoder för att analysera cerbrospinalvätska (CSF); spinal-protein och spinal-albumin. Syftet med studien var att undersöka om spinal-albumin som baseras på turbidimetri skulle kunna ersätta spinal-protein som baseras på spektrofotometri. Denna används idag som rutinanalys på laboratoriet för klinisk kemi på S Södra Älvsborgs sjukhus (SÄS). 35 prover analyserades, tagna från det CSF -prover som ankom till SÄS. Varje prov kördes två gånger med respektive metod. Studien visade en god korrelation mellan metoderna, dock visade Bland-Altman diagram mindre spridning av värdena som erhölls med albumin-metoden. Resultaten för albumin jämfördes mot Sahlgrenska universitetssjukhus (SU) resultat, då albumin-metoden ingår i deras rutiner.   Studien visade också att de dagliga kontrollerna som kördes ej var lämpade för albumin metoden, då både innehåll och koncentration inte var anpassad till spinalprover. Kontrollerna kommer att bytas ut mot andra mer lämpade kontroller. Metoden för spinal-albumin kommer att kunna ersätta total-protein som rutinanalys av CFS.

Group A Streptococcus (GAS) is a globally disseminated pathogen that causes >500,000 deaths yearly and is ranked as ninth leading infectious cause of human mortality by the World Health Organisation. The spectrum of disease ranges from superficial infections of the skin and epithelium to invasive and systemic infections. Although the interaction of GAS with neutrophils has been extensively studied much remains to be discovered about the role of macrophages, which are the first line of defence encountered by invading pathogens. In this study, the aim was to establish a means of deriving macrophages from primary monocytes and to study both the efficiency of macrophage killing of GAS and the macrophage cell death response to GAS infection. Here, we report that monocyte-derived macrophages are able to take up and kill GAS during in vitro infection. Production of reactive oxygen species by macrophages was elicited during infection, but not nearly in as high amounts as produced by neutrophils. Investigating the type of cell death induced by GAS, markers for both apoptosis and necroptosis can be found after 8 hours of infection. These results highlight that macrophages indeed are participating in the clearance of GAS and more studies are needed to understand the roles of macrophages in early control of GAS infection.