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 51^oC 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.

In peripheral blood the composition of leukocytes vary due to diseases and therefore analysis of the cells is critical in diagnosing and monitoring various leukemias. Manual microscopy and evaluation of the leukocytes is time consuming and not always clear, and it requires educated staff with long experience to assess the cells. A reliable automatic pre-classification of the cells with the system CellaVision DM12000 is therefor of great interest to facilitate quantification and classification of leukocytes. To evaluate if the instrument’s pre-classification were affected by staining, changes of the current staining solution used in clinical assessment where made by adjustment of the phosphate buffer pH. Examined blood smears that were descendent from 10 different samples were stained in buffers with pH 6.0, 6.8 and 7.5. This study showed a high positive correlation in all of the pH-adjusted buffers with the best results in pH 6.0 and 6.8. CellaVision DM1200’s pre-classification of eosinophilic granulocytes were most affected by changes in pH, which can be explained by their strongly acidophilic granules. 

The antibiotic class β-lactams include drugs such as penicillins, cephalosporines, carbapenems and monobactams which mechanism of action is to inhibit cell-wall synthesis. Bacteria have developed several mechanisms to counter β-lactams. Bacteria can defend themselves from antibiotics by releasing enzymes that attack the antibiotic compound itself by hydrolysis, target alteration or redox reactions. Presence of antibiotics can also trigger a downregulation of genes coding for antibiotic binding proteins, as well as upregulation of proteins that serves as channel and pump proteins that ensure no accumulation of antibiotics occurs in the cytosol. The aim with the study was to investigate the presence of three plasmid-mediated genes (blaFOX, blaCIT(CMY-2) and blaMOX) coding for ampicillin resistance (pAmpC) in water isolates sampled from Helge River, Kristianstad. The detection of genes was done according to a previous optimized protocol for Real-Time PCR with SYBR™Green chemistry (duplex blaCIT(CMY-2)/blaMOX and singleplex blaFOX). The method proved not to be robust for multiplex PCR, only the singelplex for the gene blaFOX could produce valid results. 30 of 96 isolates were deemed as positive for the gene, whereas 27 of 79 were considered clinical relevant. Among the 27 isolates, 16 also harbored other genes for resistance (13 blaCTX-M, 2 blaOXA, 1 blaTEM and 1 blaSHV). One isolate carried on three resistancegenes (blaFOX, blaCTX-M och blaTEM). A majority of the positive isolates, 20 out of 27, were sampled near the pumpstation. The findings indicate that Helge river might be a reservoir for dissemination of antibiotic resistance genes.

Size and function of the heart are crucial for diagnose and prognoses of cardiac disease. There are multiple methods to choose between when determining these parameters. I have tried to show correlation and agreement between 3-dimensional echocardiography (RT3DE) and myocardial scintigraphy (SPECT). 25 patients were examined with RT3DE when they were referred for stress-SPECT. Of these, seven patients were excluded because the image quality was poor or the software didn’t manage to make accurate calculations although the image quality was good. The remaining 18 patients (age 62 ± 11 year, 9 women) were evaluated. The ejection fraction and end diastolic volume of the left ventricular was calculated automatically without manual corrections of the boarders with RT3DE and compered with the results from SPECT. The results for EDV between SPECT and RT3DE were 83 ± 59 ml (37-291 ml) and 118 ± 32 ml (75-186 ml) respectively. Pearson’s correlations coefficient between SPECT and RT3DE were r = 0.78 (P < 0,002). The biases between the methods were 35 ± 40 ml (P < 0,001). The results for EF calculated with SPECT and RT3DE were 62 ± 13 % (36-83 %) and 54 ± 8 % (36-71 %) respectively. Pearson’s correlations coefficient between the methods were r = 0.59 (P = 0,009). The bias were calculated to -7 ± 11 % (P = 0,004) for the EF. The variation between the methods were wide and no clear relationship cloud be interpreted. To visualize that the difference between the methods grow as the size became larger a trend line was put in to the difference plot. The variation between the methods emphasizes the importance of educated and experienced personnel in the evaluation of separate results no matter which analysis method chosen.

Vibrio parahaemolyticus is a marine bacterium that humans can ingest through fish and shellfish or when exposed to seawater with high concentration of the bacteria, and it can cause gastrointestinal illness. It can also cause infection in ears and wounds. V. parahaemolyticus may be pathogenic to marine organisms. Since increased carbon dioxide concentration in the atmosphere may lead to ocean acidification, which will weaken the immune systems of marine organisms, it is important to study gene expression in V. parahaemolyticus at lower pH in order to know if it is as virulent in lower pH as in the pH of today. The aim of this study is firstly to compare bacterial survival in natural seawater and artificial seawater (ASW). 4

ASW is often used for studying of marine bacteria in laboratory work. Secondly, the study aims to compare four commercial kits (TRIzol Max bacterial RNA Isolation Kit, RNeasy Mini Kit (Qiagen), NucleoSpin RNA Plus and SV Total RNA Isolation Kit (Promega)) for RNA extraction from both log-phase at the recommended bacterial counts and marine water at low bacterial counts. Results show that the bacteria survived equally well in both kind of water. As for RNA extraction, TRIzol gave the highest RNA yield but lower quality, especially when extracted from bacteria in natural marine waters. NucleoSpin gave little higher RNA yield than the Qiagen, but the quality of the RNA extracted with Qiagen was estimated as the highest compared to other kits. This together with very short extraction time, as well as better security and easier steps can be a reason to prefer Qiagen and NucleoSpin than TRIzol.

Dehydration process is an important histotechnical method of tissue samples undergoing histopathological analysis. Using different alcohols and intermedium, the samples are processed for diffusing the water out of the tissue. Impregnation with paraffin of the tissue in the later stagesis crucial for tissue stability in the remaining histotechnical methods. Sakura Tissue-Tek VIP 6 is atraditional dehydration instrument that realizes dehydrations during longer times with xylene asthe intermedium. However, LOGOS is another dehydration instrument operating with shorter times using Histolab Clear, a more environmental and cheaper intermedium.In the current study tissue samples were used from three different patient cases of each tissue:cutis, breast, intestine and prostate. Two different, new LOGOS- dehydration programs with Histolab Clear as intermedium were tested and validated in accordance with specific qualitycriteria. Parallel were tissue samples dehydrated from the same tissues in the traditional routineprogram, in order to evaluate by comparison, sections and staining quality of the new programs.Immunohistochemistry and routine staining were used for evaluation. The pathologist approved allroutine stains and disapproved some immunohistochemical analyses, depending on poorer staining and the quality of the sections.Because the amounts of the samples were insufficient in some cases, the number of the samples were determined to be expanded in order to investigate if the error occurs systematically or onlyto individual cases. Insufficient dehydration seemed to be fundamental as a cause of inferior results. Therefore, even the time of dehydration in Histolab Clear will increase by one hour, in the further validation. Optimization of dehydration time is required for approval of immunohistochemical analyzes. These analyzes are very important and definitive for the treatment of the patient, and therefore the new dehydration programs must give at least as good results as the conventional.

Copy number variation (CNV) is known to cause all von Willebrand disease (VWD) types, although the associated pathogenic mechanisms involved have not been extensively studied. Notably, in-frame CNV provides a unique opportunity to investigate how specific von Willebrand factor (VWF) domains influence the processing and packaging of the protein. Using multiplex ligation-dependent probe amplification, this study determined the extent to which CNV contributed to VWD in the Molecular and Clinical Markers for the Diagnosis and Management of Type 1 von Willebrand Disease cohort, highlighting in-frame deletions of exons 3, 4-5, 32-34, and 33-34. Heterozygous in vitro recombinant VWF expression demonstrated that, although deletion of exons 3, 32-34, and 33-34 all resulted in significant reductions in total VWF (P < .0001, P < .001, and P < .01, respectively), only deletion of exons 3 and 32-34 had a significant impact on VWF secretion (P < .0001). High-resolution microscopy of heterozygous and homozygous deletions confirmed these observations, indicating that deletion of exons 3 and 32-34 severely impaired pseudo-Weibel-Palade body (WPB) formation, whereas deletion of exons 33-34 did not, with this variant still exhibiting pseudo-WPB formation similar to wild-type VWF. In-frame deletions in VWD, therefore, contribute to pathogenesis via moderate or severe defects in VWF biosynthesis and secretion.

Norovirus causes winter vomiting disease and is one of the commonest cause of winter illness in Sweden. The disease period generally lasts one to three days with symptoms like vomiting and/or diarrhea. To the disease burden of acute gastroenteritis worldwide, norovirus contributes with 18 %. Even though the illness is very common, the knowledge about norovirus is poor and largely unknown.The purpose of the study was to do a time series analysis, a so-called One-Step Growth analysis, of the minus-RNA concentration in cells infected with different concentrations of murine norovirus (MNV). For the detection of minus-RNA RT-qPCR was used with SYBR Green. The goal was to correlate start concentration of virus at any time with the amount of minus-RNA in the cells. At 4 and 8 hours there was an exponential connection by the initial virus concentration and minus-RNA development in the cells. The concentration of minus-RNA in the infected cells increased between 4, 8 and 24 hours. Further, the results can be interpreted as requiring 4 hours for the higher concentrations to become quantifiable, while requiring 24 hours for the lower concentrations to become quantifiable.

Transplantation is based on a satisfactory matching of the patient and donor genes for Human Leukocyte Antigen (HLA), which increases the chance of a successful transplantation. HLA gives individual cell surface markers. The Major Histocompatibility Complex (MHC) region, encoding HLA in humans, is the most polymorphic in the human genome. The genes are located on chromosome six and consists of 200 genes. Those genes encode protein products essential for the acquired immune system. MHC molecule’s role is to represent foreign substance for B- and T-lymphocytes. MHC is an important system as it contributes to the activation of the immune system to combat viruses, bacteria, parasites and cancer cells. HLA-typing is determined through certain antigens in the HLA system. The classical transplantation antigens are HLA-A, -B, -C, -DR, -DQ and -DP. By amplifying the DNA with sequence specific primers in the Polymerase Chain Reaction (PCR), the amplicons can be detected and alleles present in the patient genome can be determined. The purpose of this study was to compare occurrence of non-specific DNA binding using master mix where Ampli Taq DNA polymerase is added and master mix with polymerase included in the PCR. Samples from 16 patients were tested with both master mix- solutions. The analyses were performed with primer plates for HLA-A, HLA-B and HLA-DRP1. The results showed that the master mix with Taq polymerase included should be applied, because it gave clearer specific band, better image quality and gave weaker and approximately 30% fewer non- specific DNA binding compared to the master mix with added Taq polymerase.

Type 1 diabetes (T1D) is regarded as an autoimmune disease. Beta cells, which produces insulin in pancreas are attacked by islet cell antibodies (ICA). This leads to gradual destruction of the beta cell, which in turn cause high level of glucose in the blood because the regulator "insulin" has disappeared. In that case the patient needs to be treated lifelong with insulin. It has been shown that the ICA reactivity consisting of reactivities against different autoantigens such as: insulin autoantigen (IAA), glutamic acid autoantigen (GAD), insulinoma antigen-2 autoantigen (IA-2) and most likely also zinc transporter autoantigen (ZnT8). Determination of ICA in serum samples is important for the classification of diabetes, prediction of T1D and the development of autoimmune therapies.

Nowadays screening of ICA is performed with ”Medipan ICA screen” which is a commercial enzyme- linked immunosorbent assay (ELISA). Positives samples are further analysed by ELISA with the indirect immunofluorescence method (IF) to ensure a final positive answer.

The purpose of this study was to evaluate and compare a new commercial ELISA kit ”RSR 2screen” with the Medipan ICA screen for use it in routine analysis to evalute if it has the same / higher specificity and sensitivity, and lower price compared with Medipan ICA screen.

Serum samples from a control group (n = 199) and a patient’s group diagnosed with T1D (n = 100 were analyzed with both ELISA methods. The results were statistically evaluated to set a threshold value for positivity and to evaluate the method's sensitivity and specificity. The result showed that both ELISA- methods gave the same sensitivity (93%) and specificity (97.5%) and a high concordance (98.7%) was achieved. Analytical price per sample for the RSR 2screen was 4.2% lower than for the Medipan ICA screen. RSR 2screen can be used instead of Medipan ICA screen. 

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.

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.

Duplex ultraljud på carotiskärlen utförs på patienter exempelvis vid misstänkt stroke, Transitorisk Ischemisk Attack (TIA), Amaurosis fugax, preoperativt eller som uppföljning vid en sedan tidigare känd carotisstenos. Vid duplex carotisundersökning mäts flödeshastigheter i Arteria Carotis Communis (CCA), Arteria Carotis Interna (ICA) och Arteria Carotis Externa (ECA) och över eventuella plack samt i vissa fall mäts även diametern av kärllumen. Av störst kliniskt intresse är stenos i ICA, och när det refereras till stenos är det alltså stenos i carotis interna som avses. I denna studie ingick 160 patienter som under en 8 veckors period utfört undersökningen på Fysiologiska avdelningen vid Helsingborgs lasarett. Vid datainsamlingens start undersöktes patientjournalerna 4 veckor retrospektivt samt 4 veckor framåt i tiden vilket gav en prospektiv patientgrupp. Resultaten visade på att av totalt 320 undersökta kärl innehöll 23 kärl 40-69% stenos, 27 kärl innehöll 70-99% stenos och 4 kärl hade en total ocklusion, 38 patienter var även remitterade för ett Holter-EKG. Bland de retrospektiva patienterna hade 10 personer 70-99% stenos varav tre åtgärdades kirurgiskt och en medicinskt. 7 av dessa patienter var asymtomatiska, 2 patienter visade vänstersidiga stroke symtom och 1 patient visade andra symtom. 4 av dessa patienter hade en unilateral stenos varav 3 av dessa hade på höger ICA och 1 hade på vänster ICA.

I studien ingick 81 kvinnor och 79 män, 8 av kvinnorna och 15 av männen hade en signifikant stenos, 4 av dessa män hade även 70-99% bilateral stenos. Det fanns en signifikant stenos i ett kärl tillhörande patientgrupp 50-59 år (=17), 7 st i grupp 60-69 år (n= 42), 12 st i grupp 70-79 år (n= 59) och 7 st i grupp 80-89 år (n=33). Grupp 0-49 år innehöll 6 patienter samt gruppen >90 år innehöll 3 patienter men i dessa två grupper hade inga patienter signifikant stenos.

Diabetes, hypertoni och rökning är 3 välkända riskfaktorer för stroke, och i detta arbete utlästes dessa riskfaktorer från patientjournalerna. 3 patienter beskrevs som rökare, 41 hade hypertoni och 25 var diabetiker. 

Prostatacancer är idag den vanligaste cancerformen i Sverige, och drygt 10 000 män årligen blir diagnosticerade med sjukdomen. En obotlig variant med dödlig utgång är kastrationsresistent prostatacancer som behandlas primärt med Docetaxel. Tumörcellerna utvecklar dock med tiden resistens mot cytostatikan, och det föreligger därför ett stort behov av att utveckla kombinationsbehandlingar med Docetaxel som grund. Mangan har i flera studier visat sig kunna inducera apoptos hos olika celltyper, och är därför intressant som ett möjligt komplement till Docetaxelbehandling. Syftet med studien var att analysera apoptos hos prostatacancerceller från cellinjen PC3 som exponerats för Docetaxel i kombination med mangan. Cellerna odlades och behandlades med Docetaxel i kombination med mangan under 24, 48 och 72 timmar. Analys av apoptos utfördes med flödescytometer efter infärgning med Annexin V och propidiumjodid. Beroende på cellernas tillstånd färgas de in i olika kombinationer och kan därmed detekteras som tidigt respektive sent apoptotiska, eller som nekrotiska. Resultaten visade på en ökning av apoptos hos kombinationsbehandlade celler, men ingen signifikant skillnad förelåg i jämförelse med obehandlade celler. Ytterligare försök bör därför upprepas med ökad koncentration av Docetaxel i kombination med olika koncentrationer av mangan, och även med andra prostatacancer cellinjer såsom DU145 och LnCaP. Dessutom bör analyser av apoptotiska markörer genomföras, i syfte att bekräfta apoptos.

Flödescytometri är en metod som analyserar cellsuspensioner genom att låta cellerna passera genom en laserstråle en och en. Cellernas påverkan på ljuset orsakar scatter vilket ger information om granularitet och cellstorlek. Lasern kan även excitera fluorescerande färger som kan vara inbundna direkt till eller till cellens inre strukturer. Celler som plottas ut på samma plats i ett flödescytometriskt scatterdiagram, som till exempel blodets B- och Tlymfocyter, kan åtskiljas med fluorescenskonjugerade antikroppar som fäster på cellernas unika CD markörer En flödescytometrisk laboration utarbetades med syfte att kunna användas för undervisning av studenter vid Högskolan Kristianstads biomedicinska analytikerprogram.

Aspekterna användarvänlighet och ekonomi beaktades därför också. Av de protokoll som

jämfördes erhöll lyseringsbuffert enligt Ballesteros et al. (2014) tillsammans med PBS med 2

% BSA som infärgningsbuffert tillräckliga cellpopulationer och antikroppsinbindning för att

kunna avläsas av en bma-student. Detta protokoll hade alla aspekter bra för en

studentlaboration och även var mest ekonomiska alternativet.

In the adult retina, ramifying microglia interact with the outer plexiform layer (OPL) monitoring the synaptic integrity between photoreceptors and post-synaptic target cells. Microglia are reactive during photoreceptor diseases, but their disease-related function(s) are not fully understood. Retinal explant cultures are model systems used to study degenerative events including photoreceptor degeneration and gliosis. Our culture paradigm, with adult porcine retinas subjected to coculture with human A-retinal pigment epithelia-19 (ARPE) cells, is an experimental approach resulting in improved photoreceptor survival and reduced gliosis. Under the in vitro pathological conditions with photoreceptor degeneration, reactive Iba1-and CD11b-immunoreactive microglia and their processes positioned in proximity with the OPL and among photoreceptor outer segments. Coculture for 3 days with ARPE-cells resulted in a significantly increased density of microglia at the OPL. After 5 days of culture, the density of microglia at the OPL was similar between coculture and control specimens. Electron microscopy revealed the presence of two subtypes of microglia: one exhibiting a dark nucleus and cytosol with dilated endoplasmic reticulum, vacuoles, endosomes and mitochondrial variations. This subtype localized close to synaptic structures in the OPL. The other subtype appeared as pale phagocytic microglia localized among degenerating outer segments. The Iba1-and CD11b-immunoreactive microglia in degenerating retina may be of two separate subtypes, which differ in localization, subcellular morphology and perhaps function.

Typ 1 diabetes (T1D) är en autoimmunsjukdom som ökar för varje år och är den vanligaste typen avdiabetes bland barn och unga (98 %). Vid T1D bildar kroppen autoantikroppar mot beta celler som finns i bukspottskörteln. Betacellerna förstörs i en immunologisk process och då kan inte cellerna producera hormonet insulin. Följderna blir att individen inte kan tillgodogöra glukos i blodet. De autoantikroppar som har identifierats är mot de Langerhanska ö-cellerna är följande: ö-cellantikroppar (Islet Cell Antibodies, ICA), insulinantikroppar (Insulin Auto Antibody, IAA), glutaminsyradekarboxylas-antikroppar (glutamic acid decarboxylase antibodies, GADA), tyrosinfosfatas-liknande proteinet IA-2 antikroppar (tyrosine phosphatase-related islet antigen 2 antibodies, IA-2A) samt zinktransportör 8 antikroppar (zinc transporter 8 autoantibody, ZnT8A).

I den här studien jämfördes två kommersiella ELISA kit för detektion av autoantikroppar i serum. RSR-2screen islet cell autoantibody (ELISA-kit, RSR, Cardiff, UK) identifierar IA-2 och GAD antikroppar och RSR-3screen islet cell autoantibody identifierar (ELISA-kit, RSR,) IA-2, GAD och ZnT8 antikroppar. I studien deltog en kontrollgrupp (n=100) och en grupp av nyligen diagnostiserade T1D-patienter (n=50). Resultaten för studien behandlades statistiskt för att bestämma ett positivt tröskelvärde för båda kiten (RSR-2screen: 2,7 Units/mL och RSR-3screen:15 Units/mL). Vid specificiteten 97,5% gav RSR-2screen sensitivitet 92 % och RSR-3screen sensitiviteten 94 %. Eftersom RSR-3screen gav högre sensitivitet drogs slutsatsen att den rekommenderas i första hand för screening i en frisk population samt bland individer som löper ökad risk att utveckla T1D.

Prostate cancer is the sixth most common cancer type in the world, and the third most common cancer type among men. The different types of treatments that are available today do not usually cure the disease. It is therefore important to develop better treatment methods. It has previously been stated that manganese can cause apoptosis in different cell types. It provides an opportunity to use manganese to inhibit cancer and it is therefore important to know which apoptotic markers that are involved. The purpose of the project was to investigate whether an increase in the apoptotic markers cleaved Kaspas-3 and cleaved PARP after manganese treatment of prostate cancer cells. Thereafter, it can determine whether manganese treatment has caused cell death by inducing apoptosis. During the project prostate cancer cells (PC3-cells) were cultivated, then treated with 200 µM manganese for 6, 24 and 48 hours. The amount of the proteins cleaved Kaspas-3 and cleaved PARP was measured by using a sandwich ELISA kit. A clear gradual increase of apoptotic markers with incubation time was expected. The expected result was not obtained, for cleaved Kaspase-3 manganese treatment even decreased the concentrations. There may have been problems with the performance of the analysis such as poor lysis of the cells or uneven growth of the cells. More studies are required to investigate this and other potential apoptotic markers.

För att diagnostisera malignt melanom och dess undergrupper används immunhistokemi för att färga in celler som uttrycker specifika protein. Särskilt desmoplastiska melanom kan initialt felbedömas baserat på utseendet som ärr, och det är därför viktigt att ha sensitiva antikroppar för att diagnostisera dem. Elva arkiverade patientprover (varav tre epiteloida melanom, fyra spolcelliga, tre desmoplastiska och ett akralt lentiginöst) färgades in med antikroppar mot CK18, HMB-45, Melan-A, S100, SOX10 och synaptophysin. Alla prover var negativa för CK18, nio var positiva för HMB-45 och Melan-A (de negativa var båda desmoplastiska melanom) och alla var positiva för S100 och SOX10. Synaptophysin var positiv i alla epiteloida melanom och det akralt lentiginösa melanomet, två av de fyra spolcelliga melanomen och negativ i alla desmoplastiska melanom. Fördelarna med SOX10, som tidigare studier i ämnet har påvisat, observerades inte i denna studie, troligen på grund av begränsningen i patientmaterial. Trots det verkar SOX10 vara ett användbart tillägg i melanompanelen, eller kanske till och med kan ersätta S100, baserat på tidigare studier.

Individuals that suffer from homozygote Familiar Hyperkolesterolemia (FH), has increased amounts of Low Density Lipoproteins (LDL) which leads to a higher risk of cardiovascular diseases. Treatment of these individuals can be achieved by extracorporeal elimination of LDL using specific columns. The aim of this study was to evaluate different agarose-modified adsorbents ability to adsorb LDL from human plasma. The adsorbents (DALI, Zetarose, sulphonated Zetarose and taurine immobilized onto Zetarose) were incubated for 60 minutes with human plasma diluted with PBS, in a ratio of 1:5 between the matrix and the plasma during rotation with a rotator. After incubation the samples were centrifuged and the LDL content was determined in both the supernatant and the pellet. The amount proteins adsorbed were assayed by eluting the pellets. LDL was determined indirectly using Friedwalds equation; LDL= Total cholesterol (TC) - High density lipoprotein (HDL)-(0,45x Triglycerides (TG). The values of TC and TG in the sample were determined enzymatically, whilst HDL was quantified as TC after LDL-precipitation by dextran sulfate. The results clearly show that DALI has good adsorption capacity, but none of the modified Zetaroses shows any capacity to absorb LDL from human plasma. Desorption of the adsorbents using SDS gave higher amounts of eluated protein compared to NaCl elution, indicating hydrofobic proteins. However, the methods used in this study could be used to evaluate new adsorbents for LDL-elimination applications in patients with chronic hyperlipemia.

Venous thromboembolism that cause blood clotting in blood vessels, prevent blood circulation, depending on changes in one or more of the coagulation factors II, VII, IX and X. Patients who have had a blood clot or cardiovascular diseases are treated with oral anti-vitamin K (Warfarin^®) to reducing and prevent relapse. Warfarin is also used as a preventive treatment before the disease. An overdose of Warfarin^® may cause bleeding-complications and low dose cause blood clotting. The dosage of the drug is controlled by measuring prothrombin in plasma. The aim of this study was to investigate if prothrombin-complex value changes due to re-spinning and re-analysis after six hours. Fitty whole blood samples from warfarin-treated patients were divided into three subgroups, those with protrombinkomplex-values of 2-4 (n=20), >4 (n=15) and <2 (n=15). The samples were centrifugated and measured (Method A), re-centrifugated and measured (Method B) or re-analysed after six hours (Method C). All results were compared in a Bland-Altman plot as follows: Method B vs. Method A and Method C vs. Method A. The scatter graph yielded a strong correlation between Method A and Method B (R²=0.9984) and Method A and Methods C (R²=0.9977). The results from t-test showed a significance level (p<0.001) for both analyses (statistical significance=p<0.05). In this study we showed that prothrombin complex value ware stable after re-centrifugation and re-measurement after six hours. Statistical calculations yielded a strong correlation between the methods (A, B, C), and there was no significance difference between the methods.

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.

von Willebrand factor (VWF) levels in healthy individuals and in patients with type 1 von Willebrand disease (VWD) are influenced by genetic variation in several genes, for example, VWF, ABO and STXBP5. Here, we comprehensively screen for STXBP5 variants and investigate their association with type 1 VWD in Swedish patients and controls. The coding region of the STXBP5 gene was re-sequenced in 107 type 1 VWD patients and the detected variants were genotyped in the type 1 VWD population and a Swedish control population (464 individuals). The functional effects of missense alleles were predicted in silico and the pattern of genetic variation in STXBP5 was analysed. Re-sequencing of 107 type 1 VWD patients identified three missense and three synonymous variants in the coding sequence of STXBP5. The low-frequency missense variants rs144099092 (0.005) and rs148830578 (0.029) were predicted to be damaging, but were not accumulated in patients. No other rare candidate mutations were detected. STXBP5 showed a high level of linkage disequilibrium and a low overall nucleotide diversity of π = 3.2 × 10^-4 indicating intolerance to variants affecting protein function. Three previously type 1 VWD-associated single nucleotide polymorphisms were located on one haplotype that showed an increased frequency in patients versus controls. No differences in messenger ribonucleic acid abundance among haplotypes could be found using Genotype-Tissue Expression project data. In conclusion, a haplotype containing the STXBP5 Asn436Ser (rs1039084) mutation is associated with type 1 VWD and no rare STXBP5 mutations contribute to type 1 VWD in the Swedish population.

Electroneurography is used to examine a suspected entrapmentneuropathy in peroneus communis (PC), where an electric stimulus enables the evaluation of nerve conduction velocity, muscle response amplitude and latency. If registration from the extensor digitorum brevis (EDB) provides unreliable results, the registration can be made from tibialis anterior (TA). Currently there are no normal values available in our laboratory and no standard method regarding the registration on TA. The purpose of this study was therefore to retrieve normal values for this registration and to develop and establish a method, and also compare the different registration sites, to examine the side differences from the registrations on TA, and how the height affected the latency. It was also examined if so called F-waves could be recorded from TA, and if so, determine the response rate and latency. 22 participants between 23-59 years an 154-190 cm were examined. TA was examined with the active registration electrode on the site where the muscle was the largest and the reference electrode on the ankle. Stimulations were made on lateral poplitea fossa and 110 mm lower on distal caput fibula. EDB were examined according to established methods. Normal values for the registration on TA were 2,2-5,4 mV regarding amplitude, 55-73 m/s regarding nerve conduction velocity and 3,8-5,9 ms regarding latency. Side differences were 0-1,4 mV regarding amplitude, 0-8 m/s regarding nerve conduction velocity and 0-0,8 ms regarding latency. The calculated limits show that it only takes small side differences to have a clinical significance. The method gave equivalent results to previous studies. 23 % of the latency could be explained by height. The comparing of the nerve conduction velocity from the different registrations showed a significant statistical, but not necessarily clinical, difference, with the bias 5 m/s. F-waves were retrieved from all participants with a response rate of 94-100 %. 41 % of the F-wave latency could be explained by height.

BACKGROUND: Inversions involving intron 22 (Inv22) of F8 are detected in approximately 45% of all severe hemophilia A patients. Diagnosis is complicated by the large size of the ~9.5 kb int22h repeated sequence which generates the inversions. Methods such as long-range PCR and inverse-shifting PCR are currently used diagnostically, but suffer from low PCR efficiencies and are difficult to standardize.

OBJECTIVES: To design and validate a sensitive and robust assay for the detection of F8 int22h inversions.

METHODS: Digital droplet PCR using mile-post assays was used to investigate archival DNA samples.

RESULTS: The detection of linkage as a function of physical distance between loci was investigated using an anchor locus and mile-post loci located at 1, 6, 12 and 15 kb distances from the anchor locus. The proportion of linked molecules decreased with increasing distance between loci and showed 30-40% linked molecules for loci 12-15 kb apart. Mile-post assays specific for wild type and Inv22 type 1 and 2 chromosomes were then designed and optimized. All three assays showed high specificities and sensitivities, with coefficients of variation < 5% for all assays. Analysis of 106 patients and 20 carrier mothers showed complete concordance with previously known mutation status. The analysis demonstrated the robustness of the assays versus input DNA concentration (6 ng and higher) and level of fragmentation.

CONCLUSIONS: Digital droplet PCR and mile-post assays can be used to detect F8 int22h inversions. The assay systems are technically simple to perform, highly efficient and robust.

BACKGROUND: F8 int1h inversions (Inv1) are detected in 1-2% of severe hemophilia A (HA) patients. Long-range polymerase chain reaction (PCR) and inverse-shifting PCR have been used to diagnose these inversions.

OBJECTIVES: To design and validate a sensitive and robust assay for detection of F8 Inv1 inversions.

METHODS: Archival DNA samples were investigated using mile-post assays and droplet digital PCR.

RESULTS: Mile-post assays for Inv1 showing high specificities and sensitivities were designed and optimized. Analysis of four patients, two carrier mothers and 40 healthy controls showed concordance with known mutation status with one exception. One patient had a duplication involving exons 2-22 of the F8 gene instead of an Inv1 mutation. DNA mixtures with different proportions of wild type and Inv1 DNA correlated well with the observed relative linkage for both wild type and Inv1 assays and estimated the limit of detection of these assays to 2% of the rare chromosome.

CONCLUSIONS: The mile-post strategy has several inherent control systems. The absolute counting of target molecules by both assays enables determination of template quantity, detection of copy number variants and rare variants occurring in primer and probe annealing sites and estimation of DNA integrity through the observed linkage. The presented Inv1 mile-post analysis offers sensitive and robust detection and quantification of the F8 int1h inversions and other rearrangements involving intron 1 in patients and their mothers.

Background: The occurrence of mosaicism in hemophilia A (HA) has been investigated in several studies using different detection methods. Objectives: To characterize and compare the ability of AmpliSeq/Ion Torrent sequencing and droplet digital polymerase chain reaction (ddPCR) for mosaic detection in HA. Methods: Ion Torrent sequencing and ddPCR were used to analyze 20 healthy males and 16 mothers of sporadic HA patients. Results: An error-rate map over all coding positions and all positions reported as mutated in the F8-specific mutation database was produced. The sequencing produced a mean read depth of >1500X where >97% of positions were covered by >100 reads. Higher error frequencies were observed in positions with A or T as reference allele and in positions surrounded on both sides with C or G. Seventeen of 9319 positions had a mean substitution error frequency >1%. The ability to identify low-level mosaicism was determined primarily by read depth and error rate of each specific position. Limit of detection (LOD) was <1% for 97% of positions with substitutions and 90% of indel positions. The positions with LOD >1% require repeated testing and mononucleotide repeats with more than four repeat units need an alternative analysis strategy. Mosaicism was detected in 1 of 16 mothers and confirmed using ddPCR. Conclusions: Deep sequencing using an AmpliSeq/Ion Torrent strategy allows for simultaneous identification of disease-causing mutations in patients and mosaicism in mothers. ddPCR has high sensitivity but is hampered by the need for mutationspecific design.

Inom klinisk neuropatologisk (NP) diagnostik analyseras hjärnvävnad tillvaratagen vid obduktion, för att säkerställa vilken typ av demens en patient lidit av. Vid en NP undersökning undersöker man bland annat hur olika proteinuttryck ser ut i olika delar av hjärnan. Proteinuttryck i vävnad kan, med immunohistokemiska metoder (IHC), märkas in och göras synligt för mikroskopiundersökning. IHC resultat tillsammans med kliniskt ställd diagnos, makroskopiskt synlig degeneration och vävnadsförändringar påvisade med mikroskopi läggs samman vid NP-diagnostik. Syftet med denna studie var att undersöka hur tillförlitlig Tissue Microarray (TMA) metoden är för att analysera specifika proteinuttryck i hjärnvävnad från patienter med Frontotemporal degeneration (FTD) och Alzheimers sjukdom (AD) i förhållande till helsektionssnitt. Med TMA teknologin tas med en punktionsnål vävnadsstansar från ett flertal vävnadsklossar vilka komprimeras i en enda kloss. I studien extraherades fyra stansar(1 mm) per kloss ur vävnadsklossar från 33 patienter, en kloss per patient .Stansarna överfördes till en tom paraffinkloss med förgjorda cylindriska hål. Därefter snittades dessa TMA-klossar och snitten färgades in med fyra antikroppar, TAR DNA binding Protein (TDP-43), Fused in Sarcoma (FUS), ubiquitin och β-amyloid. Infärgningsresultaten från TMA-snitten jämfördes med resultaten från helsektionssnitten. Resultaten visade en hög (> 90 %) överensstämmelse efter analys av två, tre respektive fyra stansar/kloss för TDP-43, FUS och ubiquitin. β-amyloid visade en låg (< 90 %) överensstämmelse oberoende av antal stansar som analyserats. En procentuell ökning i överenstämmelse sågs däremot för alla antikroppar vid analys av ökat antal stansar. Studien visade att TMA kan användas inom demensforskning för att analysera specifika proteinuttryck i hjärnvävnad från patienter med FTD och AD. Antal stansar per kloss och patient är dock beroende på vilka antikroppar som ska användas. TMA är ett välbehövt verktyg vid demensforskning inom neuropatologi. Förutom att det förenklar genomförandet av stora studier och sparar pengar och tid, så sparas även värdefull vävnad som kan användas för fler analyser.

Glycated hemoglobin, HbA1c, is an indication of average long-term glucose. HbA1c is used as a diagnostic method for diabetes but also as a follow-up for diagnosed diabetics. Follow-ups shows how well a diabetic relates to diet but also medication. The information of the patient's HbA1c value plays an important factor in further treatments. The analysis method for HbA1c is not standardized, which has resulted in several analysis methods developed for HbA1c.

The purposes of this study were to investigate whether an enzymatic, Direct enzymatic HbA1c or immunological assay method, Hemoglobin A1c, can solve the problem of hemoglobin variations in the analysis of HbA1c, which is currently analyzed by HPLC as a routine at the clinical chemistry laboratory in Västerås

The implementation was performed on two instruments, TOSOH G7 and AU 680. The values from TOSOH (HPLC) were used for comparison of the two analysis methods applied on the AU680. Preparation and treatment, such as hemolysis, occurred before putting the samples into the AU680 instrument.

The result showed that the Hemoglobin A1c assay method was well-matched with HPLC assay (R2 = 0.98) in comparison to that of the Direct enzymatic HbA1c assay method (R2 = 0.86). Similar results could be observed for Hemoglobin A1c (R2 = 0.98) and Direct Enzyme HbA1c (R2 = 0.95) when only samples from patients with hemoglobin variants were analyzed (n=10). Solely analysis of hemoglobin variants with Hemoglobin A1c showed a boundary case for a significant difference compared to HPLC analysis (P = 0.051; n=134).

Incorrect reagents were obtained from the reagent manufacturer in the case of Direct enzymatic. This can explain the results obtained. Hemoglobin A1c should also be investigated with more extensive test materials for possible standardization in the routine of KKTM in Västerås.

Plasma levels of von Willebrand factor (VWF) vary considerably in the general population and this variation has been linked to several genetic and environmental factors. Genetic factors include 2 common single nucleotide variants (SNVs) located in VWF, rs1063856 (c.2365A>G) and rs1063857 (c.2385T>C), although to date the mechanistic basis for their association with VWF level is unknown. Using genotypic/phenotypic information from a European healthy control population, in vitro analyses of recombinant VWF expressing both SNVs, and in vivo murine models, this study determined the precise nature of their association with VWF level and investigated the mechanism(s) involved. Possession of either SNV corresponded with a significant increase in plasma VWF in healthy controls (P < .0001). In vitro expression confirmed this observation and highlighted an independent effect for each SNV (P < .0001 and P < .01, respectively), despite close proximity and strong linkage disequilibrium between them both. The influence of c.2365A>G on VWF levels was also confirmed in vivo. This increase in VWF protein corresponded to an increase in VWF messenger RNA (mRNA) resulting, in part, from prolonged mRNA half-life. In addition, coinheritance of both SNVs was associated with a lower VWF propeptide-to-VWF antigen ratio in healthy controls (P < .05) and a longer VWF half-life in VWF knockout mice (P < .0001). Both SNVs therefore directly increase VWF plasma levels through a combined influence on VWF biosynthesis and clearance, and may have an impact on disease phenotype in both hemostatic and thrombotic disorders.

Malaria is a parasitic disease known worldwide. In 2015 the disease caused 438000 deaths of which 90% occurred in the African region.

The parasites destroy the erythrocytes in the human body and are transmitted by the Anopheles mosquitoes. There are five species of the genus Plasmodium that causes malaria. The species P. falciparum causes the most deaths and therefore most diagnostics are about detecting this species.

Malaria occurs approximately a week after the infecting bite. The symptoms are often varying and nonspecific and hence complicate the diagnosis of the disease.

Thick and thin blood film microscopy examination (Pheripheral blood smears) is the golden standard for malaria diagnosis. However, to be able to make quicker and more reliable diagnoses new methods need to be developed. Even though there are effective drugs against malaria, delayed diagnoses and treatment are major causes of death in many countries. Knowledge and resources are primarily lacking in the endemic areas where it is needed the most. There are many criteria for a method that should be fulfilled; such as sensitivity, rapidness, accuracy and cost-effectiveness. The method needs to be easy to use in the actual environment and it has to have potential for the future. This is a comparative study of microscopy of thick and thin blood smears (PBS), quantitative buffy coat (QBC), polymerase chain reaction (PCR), cell-microarray chip, loop-mediated isothermal amplification (LAMP) and rapid diagnostic tests (RDTs).

The conclusion that can be drawn from this study is that more work needs to be done before a new method can replace conventional microscopy. In the endemic areas the aspects of cost and simplicity are important to take into consideration. Among the methods in this study, The LAMP seems to be the one with the most potential if it continues to develop in the future.

Heart failure is a term for a group of complex symtoms characterized by reduced heart function. One of these syndromes, referred to as heart failure with preserved ejection fraction (HFpEF), has increased in prevalence compared to other types of heart failures during the recent years. A concern is the difficulty in diagnosing patients with HFpEF, since current tools are considered insufficient. The aim of this thesis was to examine Peak Oxygen Uptake (VO₂peak) in relation to Total Heart Volume (THV) among heart failure patients with preserved ejection fraction (HFpEF, EF >40 %) compared to healthy controls. THV was acquired by delineating images acquired using cardiovascular magnetic resonance imaging, while VO₂peak was measured in oxygen curves acquired from cardiopulmonary exercise tests. Ratios were calculated by dividing VO₂peak with THV. In order to determine if blood hemoglobin concentration (b-Hb) could affect the ratio, ratios were adjusted to b-Hb using an adjusting factor. Mean THV was nearly 250 ml larger in HFpEF patients compared to the controls. Patients’ mean VO₂peak was more than 1000 ml lower compared to the controls. Mean VO₂peak/THV ratio calculated for the patients were less than half of that calculated for the controls. Adjusting the ratio to b-Hb did not affect the ratios significantly. The study was limited by the size of the test group, but the findings suggest that a VO₂peak/THV ratio can be used to separate HFpEF patients from healthy controls.

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.

Deposition of amyloid beta (Aβ) in the extracellular environment are associated to some severe diseases, like Alzheimer’s disease and age-related macular degeneration (AMD).

Amyloids are characterized by insolubility, toxicity towards neuron and are there-for damaging to tissues. AMD is the primary cause of blindness and irreversible loss of central vision through manifestation of Aβ in the macula. In AMD, Aβ drives an inflammatory action that degenerates the retinal pigment epithelium and cause atrophy of photoreceptors.

Today ~150 million people live with AMD where many find difficulties performing everyday tasks due to loss of sharp vision.

Congo red is a gold standard for visualizing amyloids in vitro and the pathogenesis caused by amyloids can be analyzed by immunofluorescence and immunohistochemistry.

The purpose of this study was to show the presence of amyloids relating to cell death in pig retina, show the pathogenesis caused by amyloids by using immunofluorescence and immunohistochemistry, and investigate whether there is correlation between amyloids and cell death.

The result showed that amyloids were present in the retina and caused cell death and gathering of aggresomes.

Amyloids and the caused pathology could be visualized in the outer layer of the retina.

Kommunalt avloppsvatten innehåller stora mängder föroreningar bland annat organiskt material som påverkar organismerna i naturen. Reningsverken runtom i Sverige arbetar ständigt för att minimera utsläppen av föroreningar. Det är viktigt att kontinuerligt analysera vattnets kvalité för att kontrollera reningsverkets funktion. Total Organic Carbon (TOC) och Chemical Oxygen Demand (COD) är två analysmetoder som används för att mäta organisk halt i avloppsvatten. Från COD-analysen uppkommer farligt avfall som innehåller kvicksilver varför analysmetoden enligt EU-direktiv ska fasas ut. TOC-analysen föreslås som ersättningsmetod. Syftet med rapporten är att presentera omräkningsfaktorer mellan de två metoderna. Ett stort antal mätdata, erhållna från tidigare utförda analyser vid Centrala reningsverket i Kristianstad, utgör grunden för beräkningarna. Resultaten visar en spridning av omräkningsfaktorerna för olika inkommande avloppsvatten medan variationen är mindre för utgående avloppsvatten. För inkommande vatten gjordes tre grupperingar av reningsverk och omräkningsfaktorn mellan COD/TOC bestämdes till 3,3, 3,8 respektive 4,2. För utgående vatten gjordes två grupperingar och omräkningsfaktorn bestämdes till 3,0 respektive 3,6. För att understryka reningsverkets resultat vid rapporteringen till olika myndigheter har ett nytt kontrollkort för TOC-analysen tagits fram.

Läkemedelsverket gav i september 2015 ut en rapport med titeln ”Miljöindikatorer inom ramen för nationella läkemedelsstrategin (NLS)”. Rapporten indikerar bland annat stora kvalitetsbrister i tidigare utförda analyser av läkemedel i miljön under åren 2002-2013, men betonar också vikten av att samordna framtida nationella läkemedelsanalyser för att bättre använda våra offentliga resurser. I rapporten skriver man följande: ”Allra högst prioriterad ansåg arbetsgruppen att indikatorn ”mäta halter av läkemedelssubstanser i miljö” vara. Detta beroende på att det utöver att det är av stor vikt att följa utvecklingen av läkemedelsrester i miljön över tid för att utvärdera effekten av genomförda insatser, så ansåg arbetsgruppen att det finns stora möjligheter att optimera användningen av de offentliga resurserna genom en bättre samordning av mätningar i miljön. Många mätningar har gjorts historiskt av olika offentliga aktörer utan någon samordning.” Mot bakgrund av detta, och som svar på ett nationellt behov, åtog sig Högskolan Kristianstad (HKR) genom Ola Svahn och Erland Björklund, båda verksamma vid MoLab, Krinova Incubator and Science Park i Kristianstad, att leda denna interkalibreringsstudie för läkemedelsanalys i miljö-vattenprover av olika typ. Arbetet har skett på uppdrag av Havs- och Vattenmyndigheten som finansierat arbetet via bidrag ur havs- och vattenmiljöanslaget och i samarbete med fyra andra analyslaboratorier vid Umeå Universitet (UU), Sveriges Lantbruksuniversitet Uppsala (SLU), Svenska miljöinstitutet Stockholm (IVL) samt Aarhus Universitet Danmark (AU). Total deltog därmed 5 laboratorier, alla med tidigare erfarenhet av läkemedelsanalys i miljöprover.

Abstract

Background: To prevent bacteria from forming biofilm on medical devices is an important challenge.Biofilm formation is associated with decreased susceptibility to antibioticsand the development of antibiotic resistance. The problem has to be solved by both preventing the formation of biofilm and development of new antibiotics directed to the formation of biofilm. Antibacterial coating on medical devices is a preventive action to decrease the risk of catheter related infections. The Bactiguard coating is one example, and has been clinically proven to reduce infections. But if the Bactiguard coating affects formation of microbal biofilm or production of extracellular polymeric substances (EPS) is not yet fully understood.

Methods: The amount of biofilm formed on coated and uncoated surface was compared by staining of EPS. Three different bacterial species were cultured separately and together.

Results: All three species of bacteria did form biofilm on Bactiguard coated surface. However, there was significantly less bacterial growth on the Bactiguard coated microtiter plates compared to the uncoated. Quantification of EPS was not successful. Colonization of S. epidermidis appeared differently on Bactiguard coated microscope slides after 4-8 h of culture.

Discussion: The antibacterial effects of Bactiguard coated catheters seem to be more connected to inhibited adherence of bacteria to the surface.  The coating has to be optimized for further study on microtiterplates.

Heart failure is a cardiovascular disease which affects more and more people in the World. Every year, approximately one million patients in the United States seek care for symptoms of heart failure. Chronic heart failure is categorized by ejection fraction, preserved or educed ejection fraction. In preserved ejection fraction it is common with a reduced filling of the left ventricle. Magnet resonance imaging (MR) is a test method which uses the hydrogen ions in the body to create an image using a magnetic field and is considered a “gold standard” to measure the heart volumes and mass. The aims of this study were to compare left ventricular volumes, mass and function in heart failure patients with preserved ejection fraction and healthy controls. The hearts of seven patients and eight healthy controls were examined in a magnetic camera at Skåne’s University Hospital in Lund. The images were analyzed by manual delineation of the left ventricle. No statistical significant difference could be demonstrated between the groups. In contrast with earlier studies this study did not show any difference in left ventricular volumes, mass and function. However, the current study is limited due to the low number of patients and controls. 

 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.

INTRODUCTION: Venous thromboembolism (VTE) is a complex disease that aggregates in families. Both acquired and genetic risk factors are important. Proper recognition and management of high-risk individuals are important.

AREAS COVERED: The genetic risk factors for VTE, the clinical consequences, and future perspectives are summarized. Classical thrombophilia i.e. factor V Leiden (rs6025), the prothrombin G20210A mutation (rs1799963), deficiencies of antithrombin, protein C, and protein S and the recent findings from genome wide association studies (GWAS), transcriptome-wide association studies (TWAS), genetic risk score (GRS), VTE candidate genes, expression studies, animal studies, studies using next generation sequencing, pathway analysis, and clinical implications are discussed.

EXPERT OPINION: Screening of inherited thrombophilia should be performed in special cases. Identification of strong risk variants might affect the management. The increasing number of genetic risk variants is likely to change management of VTE.