Avsaknad av en ”gyllene standard” och en låg sensitivitet hos förstahandsmarkören total B12 har varit de stora begränsningarna i diagnostiken av vitamin B12-brist anemier. Med syftet att förbättra diagnostiken av bristanemier vid Länssjukhuset Ryhov, Jönköping utvärderades användningen av aktivt B12 vid bristanemiutredningar och en metodverifiering för analysen genomfördes på ARCHITECT i2000SR. Sambandet mellan aktivt B12 och totalt B12 undersöktes med en korrelationsanalys för totalt B12-nivåer: låg (n=10, <148 pmol/L), gråzonen (n=10, 149–200 pmol/L) och normal (n=10, 201-250 pmol/L). För bestämning av träffsäkerheten hos aktivt B12 studerades 14 bristanemifall. Ett receiver operating characteristic (ROC) diagram skapades och arean under kurvan (AUC) beräknades. Variationskoefficient (CV) för mellanliggande- och inomserieprecision för aktivt B12 var 4,4 % respektive 2,4 %. Svag korrelation observerades i alla B12-nivåer, men sammantaget resulterade nivåerna i en medelstark korrelation (R=0,600). Aktivt B12 med ett gränsvärde på 40 pmol/L resulterade i en hög specificitet (75 %) och sensitivitet (80 %) med en AUC på 90 %. Analys av aktivt B12 på ARCHITECT i2000SR möjliggör en precis mättning av aktivt B12 i humant serum och därför är lämplig för rutinmässig användning. Träffsäkerheten hos aktivt B12 var hög och markören kan därmed rekommenderas för användning vid bristanemiutredningar vid Länssjukhuset Ryhov, Jönköping. / Diagnosis of vitamin B12-defiency anemia has been challenging due to lack of ”gold standard” and poor sensitivity of total B12 as a first line marker. With the aim of improving diagnostic of anemia investigations at county hospital Ryhov, Jönköping, we performed a method verification for active B12 on ARCHITECT i2000SR and evaluated the usefulness of active B12 in diagnosing anemia. Correlation analysis was performed using samples with different total B12 ranges: low (n=10, <148 pmol/L), grey zone (n=10, 149-200 pmol/L) and normal (n=10, 201-250 pmol/L). Receiver operating characteristic (ROC) curve was created and area under the curve (AUC) was calculated for determining accuracy of active B12 using 14 deficiency cases. Coefficient of variation (CV) for within-run and total imprecision, for active B12 was 2,4 % and 4,4 %, respectively. Weak correlation was found between B12-ranges, however a moderate correlation (R=0,600) was found including all groups. Active B12 with the cut-off value of 40 pmol/L resulted in high specificity (75%) and sensitivity (80%) and had an AUC of 90 %. Active B12 assay on ARCHITECT i2000SR allows a precise measurement of active B12 in human serum and therefore is adequate for routine use. Active B12 had high accuracy and can be recommended in anemia investigations in county hospital Ryhov, Jönköping.

Duktal bröstcancer är den vanligaste formen av invasiva brösttumörer. Graderingssystemet för bröstcancer har definierats av Elston och Ellis och är baserat på tre parametrar. En av dessa är räkning av antal mitoser på preparat färgade med Hematoxylin och Eosin (HE). Som ett komplement vid bedömning av bröstcancer analyseras Ki67, en proliferationsmarkör, med hjälp av immunhistokemi (IHC).  Fosfohiston H3 (pHH3) är ett histonprotein som finns i cellkärnan. Proteinet tros vara en specifik markör för mitos eftersom den är fosforylerad enbart under M-fasen och i slutet av G2-fasen. Syftet med studien var att utvärdera pHH3 som potentiell markör för mitos vid diagnostiseringen av duktal bröstcancer. Syftet var även att jämföra metoden med räkning av antal mitoser och Ki67-positiva celler, samt att studera den interindividuella skillnaden vid bedömningen av preparaten. Materialet bestod av 20 sektorresektat med invasiv duktal bröstcancer. Preparaten färgades med IHC och bedömdes mikroskopiskt. Celler som var positiva för pHH3 och Ki67 samt antal mitoser räknades, av tre läkare. Ett medelvärde för varje patientfall och metod beräknades från läkarnas bedömningar. Metodernas variationskoefficienter och dess medelvärden beräknades. Variationskoefficienterna uppvisade medelvärden på 0,21 för Ki67 +/- 0,10 SD, 0,33 för pHH3 +/- 0,14 SD och 0,46 för mitosräkning +/- 0,34 SD. Korrelationskoefficienterna för metoderna och respektive läkare uppvisade en spridning. Korrelationerna uppvisade medelvärden på r = 0,78 för Ki67 och pHH3, r = 0,74 för Ki67 och mitos samt r = 0,83 för pHH3 och mitos. Enligt studien verkar antipHH3 vara ett bra komplement vid bedömning av duktal bröstcancer. Dock krävs tydliga kriterier för vad som ska räknas som en pHH3-positiv cell. Intervariabiliteten verkar bli mindre med anti-pHH3 än vid räkning av mitoser, som är mer tidskrävande. Minst intervariabilitet ses vid bedömning av anti-Ki67 som en proliferationsmarkör. / Ductal carcinoma of the breast is the most common form of invasive breast tumours. The grading system for breast cancer is defined by Elston and Ellis and is based on three criterions. One of these criterions is the mitotic count in pathological sections of breast carcinomas stained with Hematoxylin Eosin. A common method often applied as a complement in diagnosis of breast carcinoma is immunohistochemical staining with use of antibodies directed against Ki67, a proliferation marker. Phosphohistone H3 is a histone protein that is located in the cell nucleus. The protein is believed to be a specific marker for mitosis since it only is phosphorylated during mitosis, and to some extent at the end of the G2-phase. The purpose of this study was to evaluate pHH3 as a potential marker for mitosis when diagnosing ductal breast cancer. The purpose was also to compare the method to mitotic figuring and the count of Ki67-positive cells, and to study the inter-individual variability when assessing the histological sections. The material consisted of 20 biopsies containing invasive ductal breast cancer. The sections were stained using IHC and all sections were evaluated microscopically. Cells positive for pHH3, Ki67 and mitotic cells were quantified, by three doctors. From the doctors results an average value was determined for each case and method. To be able to compare the methods the coefficient of variation was calculated. The average value of the coefficient of variation was determined for each method and also the standard deviation (SD). The coefficient of variation showed average values of 0,21 for Ki67 +/- 0,10 SD, 0,33 for pHH3 +/- 0,14 SD and 0,46 for mitotic figuring +/- 0,34 SD. The correlation coefficients for the methods and each doctor showed dispersion. The correlations showed average values of r = 0,78 for Ki67 and pHH3, r = 0,74 for Ki67 and mitosis and r = 0,83 for pHH3 and mitosis. According to this study it seems as though anti-pHH3 could complement the other methods. However explicit criteria which defines a threshold value of which cells should be considered pHH3-positive needs to be established. The inter-individual differences seem to decrease using antipHH3 compared with mitotic counting, which is more time consuming. Although the minimum difference can be seen when assessing anti-Ki67 as a proliferation marker.

Procalcitonin (PCT) is a precursor protein of the hormone calcitonin and is encoded by the gene Calcitonin-1. In the Blekinge Regional, P-PCT is only analyzed in Karlskrona. The analysis is performed in the department of clinical chemistry on the Alinity i-series instrument. PCT indicates bacterial infections and therefore, it is important to have a backup method for the analysis when the instrument in the city of Karlskrona is out of order. The aim of this work was to verify the analysis method of P-PCT on the instrument Alinity i-series in the city of Karlshamn. Analysis method verification means to confirm and prove that the method meets the specified requirements. Verification was performed by analyzing 35 samples with different concentration of PCT on the master instrument in Karlskrona and on “Alinity 1” and “Alinity 2” in Karlshamn. The method was compared by studying correlation coefficient and bias. The precision was measured only on “Alinity 1” which would be the master instrument in Karlshamn. Precision was measured by analyzing 25 replicates at two control levels and then was 5 replicates of each control level analyzed over 5 days. The correlation was good and no significant bias between results from Karlskrona and “Alinity 1” and between results from “Alinity 1” and “Alinity 2”. Precision on “Alinity 1” meets the requirements. The conclusion was that verification of PCT on master instrument “Alinity 1” and slave instrument “Alinity 2” was approved and the backup method for the PCT analysis in Karlshamn was verified. / Prokalcitonin (PCT) är en peptidprekursor till hormonet kalcitonin och kodas av genen kalcitonin-1 (CALC-1). PCT används som biomarkör för bakteriella infektioner och sepsis. I Region Blekinge analyseras P-PCT bara i Karlskrona. Analysen utförs på avdelningen för Klinisk kemi på instrumentet Alinity i-serie Abbot. PCT indikerar bakteriella infektioner och av denna anledning efterfrågas av flera avdelningar. Därför är det viktigt att verifiera en analysmetod för P-PCT i Karlshamn, ifall instrumenten i Karlskrona är ur funktion. Syftet med arbetet var att verifiera analysmetoden för P-PCT på instrumentet Alinity i-serie Abbot i Karlshamn. Metodverifiering innebär att bekräfta och bevisa att metoden uppfyller specificerade krav. Metodverifiering utfördes genom att analysera 35 prover med olika PCT-koncentrationer på masterinstrument ”Shrek” i Karlskrona och på ”Alinity 1” och ”Alinity 2” i Karlshamn. Metoderna jämfördes genom att studera korrelationskoefficient och bias. Inom och total serie-precision mättes bara på ”Alinity 1” som ska vara masterinstrument i Karlshamn. Inom serie-precision mättes genom att analysera 25 replikat av två kontrollnivåer och total serie-precision genom att analysera 5 replikat/dag under 5 dagar. Resultaten blev att metoden på ”Alinity 1” hade en god korrelation med metoden på masterinstrument ”Shrek” och ingen signifikant bias observerades mellan metoderna. Inom och total serie-precision på ”Alinity 1” uppfyller kraven. Metoden på ”Alinity 2” hade en god korrelation med metoden på masterinstrument ”Alinity 1” och ingen signifikant bias observerades mellan metoderna. Slutsatsen var att verifiering av P-PCT på masterinstrument ”Alinity 1” och slavinstrument ”Alinity 2” var godkänd och därmed verifierades en backupmetod för analysen P-PCT i Karlshamn.

Neuroblastoma constitutes roughly 8% of all childhood cancers where 95% of all neuroblastoma cases occur before the age of 10. The survival rate of infants and young children is very poor, which alone contributes to research novel biomarkers for classification methods, improved diagnosis and better anti-tumor therapies. The aim of this meta-analysis was to identify dysregulated miRNAs in neuroblastoma that has the potential to be used as antioncogenic biomarkers for diagnostic interventions. Additionally, explore miRNA interconnectedness on a systemic level and conversely extend the support of using miRNAs as biomarkers. A comprehensive literature search was performed within NIH-PubMed, NCBI-PMC and in the reference list of already reviewed publications, which yielded 9 eligible publications. Quality of evidence was assessed according to the guidelines adapted from MIAME, MINSEQE and MIQE. miRNet 2.0 was used to find the most significantly enriched annotations linked to neuroblastoma. A total of 251 samples (Cancer: 141; Control: 110) was reported by the 9 studies. These involved 66 dysregulated miRNAs (Up-regulated: 43; Down-regulated: 23) which was used for enrichment analysis. Four miRNAs (miR-17-5p, -92a-3p -421, -125b) were significantly linked to neuroblastoma, and associated secondary diseases; medulloblastoma (-92a-3p, -125b), bladder cancer (-17-5p, -125b), acute myeloid leukemia (-92a-3p, -125b) and cardiac hypertrophy (- 125b). miR-125b showed exceptional interconnectivity with these diseases and a multidimensional potential in neural tumorigenesis. This study showed that dysregulation and biological processes of these miRNAs were concurrent with the original studies, endorsing that these miRNAs have potential as diagnostic indicators or classifiers of such diseases. / Popular scientific summary Neuroblastoma (NB) is one of the most common types of pediatric neurological cancers in children and constitutes roughly 8% of all childhood cancer types, in which 95% of all NB cases occur before the age of 10. Even with frequent advancements in medical diagnosis and anti-tumor therapies, the current treatment options for patients with NB offers a survival rate that is very poor. This alone is a reason to pursue developing novel classification methods, improve diagnosis and research better anti-tumor therapies. Micro Ribonucleic Acids (miRNAs) are small non-coding single stranded biomolecules that have gotten a lot of attention in recent years due to their ability to regulate genes involved in various biological cancer processes, such as; tumor growth and development. miRNAs regulate these processes by altering the function of messenger RNAs (mRNAs), which are single-stranded biomolecules that resembles a piece of genetic code from the DNA of an organism cell. When these mRNAs become dysregulated, their cancer-promoting genes are disrupted which prevent them from working properly, leading to tumor regression or termination. The effect of this biological event is then objectively measured by using the miRNA as an indicator, also known as a biomarker. miRNA biomarkers have massive potential to improve various medical applications, such as; faster and more accurate diagnosis, detailed disease-classification and more precise drug trial predictions. However, a lot of individual studies have been published about the same miRNAs, which report a variation of conclusions. This makes it more difficult to determine the true nature of miRNAs. This issue can be addressed with systematic reviews and meta-analyses, which could yield additional support and give a broader picture of how miRNAs regulate different biological processes in NB. A meta-analysis is a scientific statistical process that combines the results of many research publications associated with the same scientific question and presents the best collective estimate of truth with increased precision than what could be achieved from individual studies alone. Thus, meta-analysis is an important tool in research which makes sure that the most trustworthy effect estimate can be achieved among many similar answers. The aim of this meta-analysis was to identify dysregulated miRNAs in NB that has the potential to be used as anti-cancer promoting biomarkers for diagnostic interventions. Additionally, explore how different miRNAs are connected to NB and conversely extend the support of using miRNAs as biomarkers. The end goal of this meta-analysis is to provide more reliable evidence for further research that can improve the life expectancy of NB patients in the future. In this study, 4 miRNAs (miR-17-5p, -92a-3p -421 and -125b) were identified to be significantly linked to NB, and associated secondary diseases; medulloblastoma (-92a-3p & -125b), bladder cancer (-17-5p & -125b), acute myeloid leukemia (-92a-3p & -125b) and cardiac hypertrophy (- 125b). Specifically, miR-125b showed exceptional interconnectivity for these diseases and potential to indirectly down-regulate n-Myc in NB, a gene that promote cancer cell proliferation. miR-125b was also found to be a significant sole regulator and effector of the CDX2 gene responsible for cancer cell differentiation in acute myeloid leukemia, a relationship that has been supported by other publications. This meta-analysis showed that the reported dysregulation and biological processes of these miRNAs were concurrent with the original studies, endorsing that these miRNAs have potential as diagnostic indicators or classifiers of such diseases while warranting that the gene regulatory function of miRNAs are becoming more intricate than previously thought.

Crohns sjukdom är en kronisk inflammatorisk sjukdom som påverkar hela gastrointestinala kanalen, från mun ner till ändtarmen. Sjukdomen är progressiv och innefattar olika skov av symptom, som till exempel diarré, magont, rektalblödning, viktminskning, feber och trötthet. Den exakta patofysiologiska orsaken är ej klargjord, men tros bero på olika faktorer som till exempel genetiska faktorer, miljöfaktorer samt immunologiska faktorer. Det är känt att individer med inflammatorisk tarmsjukdom (IBD) uttrycker en större population av mastceller och eosinofiler i tarmsubmukosa, och kan orsaka diverse symptom. Histamin-4 receptorer är G-proteinkopplade receptorer som har en nyckelroll i att förmedla inflammatoriska svar, och spelar en roll i immuncellsmigration till inflammerade vävnader. Syftet med projektet är att studera uttrycket av histaminreceptorn H4 på eosinofiler och mastceller i inflammerad tarmvävnad från patienter med Crohns sjukdom. Det medverkade 16 patienter, varav 8 hade Crohns sjukdom, och 8 var friska kontrollpatienter som opererats för tarmcancer. Uttrycket av eosinofiler, mastceller och Histamin-4 receptorer studerades med hjälp av immunohistokemisk metod. Resultatet visar statistisk signifikanta skillnader mellan patient och kontrollgruppen när det kommer till eosinofiler och mastceller, med ett p-värde <0,05. Det uppvisades ingen signifikant skillnad mellan grupperna när det kom till icke inmärkta celler som uttrycker histamin-4 receptorer, eosinofiler som uttrycker receptorn samt mastceller som uttrycker receptorn. Sammanfattningsvis finns det ett behov att studera detta område igen, med större urval, och möjligtvis förbättrade metoder för att komma fram till säkerställda slutsatser. / Crohn's disease (CD) is a chronic inflammatory disease that affects the entire gastrointestinal tract, from the mouth down to the rectum. The disease is progressive and involves flare-ups of symptoms such as diarrhea, abdominal pain, rectal bleeding, weight loss, fever, and fatigue. The exact pathophysiological cause is unclear but is believed to be related to various factors, including genetic, environmental, and immunological factors. It is known that individuals with inflammatory bowel disease (IBD) express a higher population of mast cells and eosinophils in the intestinal submucosa, which can cause diverse symptoms. Histamine-4 receptors are G-protein-coupled receptors that play a key role in mediating inflammatory responses and are involved in immune cell migration to inflamed tissues. The aim of the project is to study the expression of the histamine receptor H4 on eosinophil granulocytes and mast cells in inflamed intestinal tissue from patients with CD. Sixteen patients participated, including 8 with CD and 8 control patients who underwent surgery for colon cancer. The expression of eosinophils, mast cells, and histamine-4 receptors was studied using immunohistochemical methods. The results show statistically significant differences between the patient and control groups in terms of the number of eosinophils and mast cells, with a p-value <0.05. There was no significant difference between the groups in terms of un-stained cells expressing histamine-4 receptors, eosinophils and/or mast cells expressing the receptor. In conclusion, there is a need to further study this area with larger sample sizes and possibly improved methods to arrive at conclusive findings.

Type-2 diabetes has become a common disease. The complications associated with thedisease are often cardiovascular. Platelets are important components in hemostasis, one function is platelet adhesion. Platelet adhesion is the first step to form a platelet plug, together with subendothelial proteins. Blood lipids contribute to membrane structure of cells and has an important role in different signaling pathways. They have a key role in platelet activation and is associated with the changes in membrane lipids. C-reactive protein (CRP) is an acute phase protein and increases during inflammation. The physiological role is not fully known but increasing levels of CRP increases the risk for cardiovascular complications. The aim of this study was to analyze the correlation between platelet adhesion with blood lipids and high sensitivity CRP in type-2 diabetics. 69 patients with type-2 diabetes participated in this study. Lipids, CRP and platelet adhesion were analyzed, the values were used to perform correlation analysis. The results show statistically significant positive correlations between ApoA-1, HDL and platelet adhesion as well as significant negative correlations between LDL, ApoB/ApoA-1-ratio, LDL/HDL-ratio and platelet adhesion. ApoA-1 and platelet adhesion with collagen showed statistically significant correlations with r-values 0,299-0,436. ApoA-1/ApoB-quota showed statistically significant correlations with r-values from -0,492 to -0,268. Majority of correlations between totalcholesterol, ApoB, triglycerides, hsCRP and platelet adhesionshowed no statistically significant correlations. / Typ-2 diabetes har blivit en vanligare sjukdom. De komplikationer som kan uppstå vid diabetes, särskilt obehandlad diabetes, är ofta kardiovaskulära komplikationer. Trombocyter är viktiga komponenter i hemostasen, en av deras funktioner är trombocytadhesion. Trombocytadhesionen är första steget vid bildandet av en trombocytpropp, tillsammans med subendoteliala proteiner så kan trombocyterna fästa till kärlväggen och hemostasen påbörjas. Blodlipider bidrar till cellmembraners struktur, energiförvaring och har en viktig roll vid olika typer av signalering. De har en nyckelroll vid trombocytaktivering eftersom trombocytaktivering är associerad med förändringar som sker hos membranlipiderna. C-reaktivt protein (CRP) är ett akutfasprotein och ökar vid inflammation. Fysiologiska rollen är inte helt känd men stegring av CRP ökar risken för kardiovaskulära komplikationer. Syftet med denna studie är att analysera korrelationen mellan trombocytadhesion med blodlipider och högkänsligt CRP hos typ 2 diabetiker. Det medverkade 69 patienter med typ-2 diabetes i studien. Blodlipider, högkänsligt CRP och trombocytadhesion analyserades ochvärdena för respektive analyt användes för korrelationsanalyser.Resultaten visar statistiskt signifikanta positiva korrelationer mellan ApoA-1, HDL och trombocytadhesion samt signifikanta negativa korrelationer mellan LDL, ApoB/ApoA-1-kvot, LDL/HDL-kvot och trombocytadhesion. ApoA-1 och trombocytadhesion med kollagen som proteinyta uppvisade statistiskt signifikanta korrelationsintervall med r-värden på 0,299-0,436. ApoA-1/ApoB-kvoten visade statistiskt signifikanta korrelationsintervall med r-värdenmellan -0,492 till -0,268. Majoriteten av korrelationerna för variablerna: totalkolesterol, ApoB, triglycerider, högkänsligt CRP och trombocytadhesion visade inga statistiskt signifikanta korrelationer.

Standardbikarbonat innebär koncentrationen bikarbonat i plasmadelen av helblod under standardiserade förhållanden. Analysen baseras på uppmätta värden av partialtryck för koldioxid (CO2), pH, syrgasmättnad och total-hemoglobin samt en beräkning. Bikarbonat (HCO3-) har tillsammans med CO2 en central roll i syra-basregleringen där njurarna kan styra HCO3--koncentrationen och lungorna kan styra CO2-koncentrationen. Indikationer på att analysera standardbikarbonat är exempelvis misstanke om acidos eller alkalos samt hos personer med njur- och eller lungsjukdom. Syftet med examensarbetet var att studera den preanalytiska hållbarheten för vB-Standardbikarbonat för att om möjligt förlänga tiden till analys inom region Kalmar län. Analysen genomfördes på blodgasinstrumentet ABL 825 FLEX. Provmaterialet bestod av venöst helblod i mörkblå Natrium-heparin-rör från 68 blodgivare och 7 dialyspatienter. De provrör som förvarades i kylskåpstemperatur (+2 till +8 °C) analyserades efter 0, 2, 3, 4 och 5 timmars förvaring. De provrör som förvarades i rumstemperatur (+18 till +25 °C) analyserades efter 0, 1, 2, 3 och 5 timmars förvaring. Resultatet visade att hållbarheten var bättre vid förvaring i kylskåpstemperatur än vid förvaring i rumstemperatur. Vid förvaring i kylskåpstemperatur erhölls en genomsnittlig förändring på -0,14 mmol/L (-0,6 %), -0,44 mmol/L (-1,8 %), - 0,45 mmol/L (-1,8 %), -0,58 mmol/L (-2,4 %) vid respektive förvaringstid 2, 3, 4 och 5 timmar. Vid förvaring i rumstemperatur erhölls en genomsnittlig förändring på -0,36 mmol/L (-1,5 %), -0,65 mmol/L (-2,7 %), -0,98 mmol/L (-4,0 %), -1,57 mmol/L (-6,4 %) vid respektive förvaringstid 1, 2, 3 och 5 timmar. Efter genomgång av resultatet med medicinskt ansvariga läkare framkom det att två timmars förvaring i rumstemperatur och fem timmars förvaring i kylskåpstemperatur var godtagbara förvaringstider. Utifrån resultatet i denna studie föreslås därför en uppdatering av hållbarheten på analysen vB-Standardbikarbonat till två timmar i rumstemperatur och fem timmar i kylskåpstemperatur inom region Kalmar län. / Standard bicarbonate is the measured concentration of bicarbonate in the plasma-part of whole-blood under standardized conditions. The analysis is based on measurement of partial pressure of carbon dioxide, pH, oxygen saturation, and total hemoglobin as well as a calculation. Bicarbonate (HCO3-) and CO2 are particularly important components of the acid-base-regulation through the kidneys’ ability to manage the HCO3--concentration and the lungs’ ability to manage the CO2-conentration. Analysis of standard bicarbonate is indicated by, for example, a clinical suspicion of acidosis or alkalosis and in patients with renal- and or pulmonary-disease. The aim of this study was to evaluate the time from sampling to analysis regarding the analysis of vB-standard bicarbonate and if possible, upgrade the pre analytical handling within region Kalmar County. The analysis was performed with the blood gas-instrument ABL 825 FLEX. The sampling in this study consisted of venous whole-blood in dark blue sodium-heparin-test tubes from 68 blood donors and 7 dialysis patients. The test tubes were either placed at refrigerator-temperature (+2 to +8 °C) or at room-temperature (+18 to +25 °C). Test tubes stored at refrigerator-temperature were stored for 0, 2, 3, 4 and 5 hours before analysis and the test tubes stored at room-temperature were stored for 0, 1, 2, 3 and 5 hours before analysis. The results show that storage at refrigerator-temperature was better in comparison to storage at room-temperature. The mean change at refrigerator-temperature was -0,14 mmol/L (-0,6 %), -0,44 mmol/L (-1,8 %), - 0,45 mmol/L (-1,8 %), -0,58 mmol/L (-2,4 %) for each storage time 2, 3, 4 and 5 hours respectively. The mean change at room-temperature was -0,36 mmol/L (-1,5 %), -0,65 mmol/L (-2,7 %), -0,98 mmol/L (-4,0 %), -1,57 mmol/L (-6,4 %) for each storage time 1, 2, 3 and 5 hours respectively. After conversation with the medically responsible doctor about the acceptable storage time, the conclusion was that two hours at room-temperature and five hours at refrigerator-temperature were acceptable storage times. Therefore, based on the results of this study, a proposal was made to change the time of storage to two hours at room-temperature and five hours at refrigerator-temperature regarding the analysis vB-standard bicarbonate in region Kalmar County.

Aim: To study the possibility of detecting lymph node metastasis in locally advanced urinary bladder cancer (UBC) treated with radical cystectomy (RC) by using preoperative positron emission tomography/computed tomography (PET/CT) and peroperative sentinel node biopsy (SNB) technique. We also investigate the clinical significance of macrophage traits expression by cancer cells, M2-macrophage infiltration (MI) in tumor stroma and the immunohistochemical expression of biomarkers in cancer cells in relation to clinicopathologic data. Patients and Methods: We studied prospectively 122 patients with UBC, pathological stage pT1–pT4 treated with RC and pelvic lymph node dissection (PLND) during 2005–2011 at the Department of Urology, Linköping University Hospital. In the first study, we compared the results of preoperative PET/CT and conventional CT with the findings of postoperative histopathological evaluation of lymph nodes (LNs). In the second study we investigated the value of SNB technique for detecting pathological LNs during RC in patients with UBC. W also examined the significance of the primary tumor location in the bladder in predicting the site of LN metastases, and the prognostic significance of lympho-vascular invasion (LVI) and lymph node metastasis density (LNMD) on survival. In the third study, we investigate the clinical significance of macrophage infiltration (MI) in tumor stroma and macrophage-traits expression by tumor cells. In the fourth study, we investigate the cell cycle suppression proteins p53, p21, pRb, p16, p14 ARF as well as tumors proliferative protein Ki67 and DNA repair protein ERCC1 expression in cancer cells. The results were compared with clinical and pathological characteristics and outcome. Results: Prior to RC, PET/CT was used to detect LN metastasis in 54 patients. PET/CT had 41% sensitivity, 86% specificity, 58% PPV, and 76% NPV, whereas the corresponding figures for conventional CT were 41%, 89%, 64%, and 77%. SNB was performed during RC in 103 patients. A median number of 29 (range 7–68) nodes per patient were examined. SNs were detected in 83 out of 103 patients (81%). The sensitivity and specificity for detecting metastatic disease by SNB varied among LN stations, with average values of 67% -90%. LNMD or ≥8% and LVI were significantly related to shorter survival. In 103 patients, MI was high in 33% of cases, while moderate and low infiltration occurred in 42% and 25% of tumors respectively. Patients with tumors containing high and moderate compared to low MI had low rate of LN metastases (P=0.06) and improved survival (P=0.06), although not at significant level. The expression of different tumor suppression proteins was altered in 47-91% of the patients. There were no significant association between cancer specific survival (CSS) and any of the studied biomarkers. In case of altered p14ARF, ERCC1 or p21, CSS was low in case of low p53 immunostaining but increased in case of p53 accumulation, although not at a significant level, indicating a possible protective effect of p53 accumulation in these cases. Conclusion: PET/ CT provided no improvement over conventional CT in detection and localization of regional LN metastases in bladder cancer. It is possible to detect the SN but the technique is not a reliable for perioperative localization of LN metastases; however, LVI and LNMD at a cut-off level of 8% had significant prognostic values. MI in the tumor microenvironment but not CD163 expression in tumor cells seems to be synergistic with the immune response against urinary bladder cancer. Our results further indicate that altered p53 might have protective effect on survival in case of altered p14ARF, p21, or ERCC1 indicating an interaction between these biomarkers.

Rectal cancer is one of the three most common malignancies in Sweden with an annual incidence of about 2000 cases. Current treatment consists of surgical resection of the rectum including the loco-regional lymph nodes in the mesorectum. In advanced cases, neoadjuvant chemo-radiotherapy (CRT) prior to the operative treatment reduces local recurrences and enables surgery. The neoadjuvant treatment can also eradicate the tumour completely, i.e. complete response. This research project was designed to investigate the effects of preoperative radiotherapy/ CRT and analyze methods to predict response to CRT. Study I investigated the expression of the FXYD-3 protein with immunohistochemistry in rectal cancer, with or without preoperative radiotherapy. The results from the total cohort showed that, strong FXYD-3 expression was correlated to infiltrative tumour growth (p = 0.02). In the radiotherapy group, strong FXYD-3 expression was related to an unfavourable prognosis (p = 0.02). Tumours with strong FXYD-3 expression had less tumour necrosis (p = 0.02) after radiotherapy. FXYD-3 expression in the primary tumour was increased compared to normal mucosa (p=0.008). We concluded that FXYD-3 expression was a prognostic factor in patients receiving preoperative radiotherapy for rectal cancer. Study II investigated FXYD-3 expression in tumours that developed local recurrences following surgery and compared this with expression in tumours that did not develop local recurrences. There was no difference in the expression of FXYD-3 between the group that developed local recurrences and the group that did not develop local recurrences. There was no difference in survival between those with strong or weak FXYD-3 expression. We concluded that this study could not confirm the findings from study 1 i.e. that FXYD-3 expression has prognostic significance in rectal cancer. Study III was a register-based study on the incidence and effects of complete response to neoadjuvant treatment. Eight per cent of the patients with adequate CRT to achieve complete response also had a complete histological response of the luminal tumor in the resected bowel. Sixteen per cent of that group had remaining lymph node metastases in the operative specimen. Chemotherapy together with radiotherapy doubled the chance of complete response in the luminal tumour. Patients with remaining lymph node metastases had a lower survival rate compared to those without. We concluded that residual nodal involvement after neoadjuvant treatment was an important factor for reduced survival after complete response in the luminal tumour. Study IV followed up the results from the previous study by re-evaluating magnetic resonance imaging (MRI)- images in patients with complete tumour response. Two experienced MRI radiologists performed blinded re-staging of post CRT MR- images from patients with complete response in the luminal tumour. One group with lymph node metastases and another one without were studied and the results compared with the pathology reports. The sensitivity, specificity, and positive and negative predicted values for correct staging of positive lymph nodes was 37%, 84%, 70% and 57%. The size of the largest lymph node (4.5 mm, p=0.04) seemed to indicate presence of a tumour positive lymph node. We concluded that MRI couldn’t correctly stage patients for lymph node metastases in patients with complete response to CRT in the luminal tumour.

Kolesterol transporteras i blodet med hjälp av lipoproteinpartiklar. Höga nivåer av low-density lipoprotein (LDL)-kolesterol i blodet är en riskfaktor för kardiovaskulär sjukdom. Koncentrationen av LDL-kolesterol kan beräknas med hjälp av Friedewalds formel men det finns även metoder där LDL-kolesterol kan analyseras direkt. Syftet med arbetet var att verifiera metoden direkt LDL-kolesterol på analysinstrumentet Beckman Coulter AU680. Metodens inomserie- och totalimprecision analyserades. Två korrelationsstudier utfördes mellan direkt LDL-kolesterol och beräknat LDL-kolesterol, en med 43 patientprover med triglycerider < 4,5 mmol/L och en med 11 patientprover med triglycerider > 4,5 mmol/L. Friedewalds formel ska egentligen inte användas vid triglycerider > 4,5 mmol/L, men i detta fall användes formeln ändå för att utvärdera eventuella skillnader mellan metodernas resultat vid höga triglyceridkoncentrationer. Vid analys av metodens inomserieimprecision blev variationskoefficienten (CV) omkring 0,5 % vid analys av både den låga kontrollen (A1) och den höga kontrollen (A2). CV för totalimprecisionen blev 1,21 % vid analys av A1 och 1,11 % vid analys av A2. Korrelationsstudierna visade ett linjärt samband mellan metoderna men den direkta metoden gav något högre resultat vid lägre koncentrationer och något lägre resultat vid högre koncentrationer jämfört med beräknat LDL-kolesterol. Vid triglycerider > 4,5 mmol/L gav den direkta metoden betydligt högre resultat än beräknat LDL-kolesterol. Slutsatsen blev att metoden hade god precision. Överensstämmelsen mellan metodernas resultat var relativt bra för proverna med triglycerider < 4,5 mmol/L. Vid triglycerider > 4,5 mmol/L var differensen mellan metoderna stor, troligtvis på grund av falskt för låga resultat från beräknat LDL-kolesterol. / Cholesterol is transported in the blood by lipoproteins. High levels of low-density lipoprotein (LDL)-cholesterol in the blood is a risk factor for cardiovascular disease. The concentration of LDL-cholesterol can be calculated using the Friedewald formula but there are also methods that measure LDL-cholesterol directly. The aim of this study was to verify the method P-LDL-cholesterol on a Beckman Coulter AU680 analyzer. Within-run imprecision and total imprecision were analyzed. The correlation between direct LDL-cholesterol and calculated LDL-cholesterol was examined using 43 patient samples with triglyceride levels < 4,5 mmol/L and 11 patient samples with triglyceride levels > 4,5 mmol/L. The Friedewald formula is not supposed to be used on triglyceride levels > 4,5 mmol/L, but in this case the formula was used anyway to evaluate differences between the methods at high triglyceride concentrations. The coefficient of variation (CV) for the within-run imprecision was about 0,5 %, both for the low control (A1) and the high control (A2). Total imprecision had a CV of 1,21 % for A1 and 1,11 % for A2. There was a linear relationship between the methods, but the direct method gave slightly higher results at low concentrations and slightly lower results at high concentrations compared to calculated LDL-cholesterol. At triglyceride levels > 4,5 mmol/L the results from the direct method was considerably higher than calculated LDL-cholesterol. The conclusion is that the precision of the method was good. The correlation between the results from direct LDL-cholesterol and calculated LDL-cholesterol was relatively high for samples with triglyceride levels < 4,5 mmol/L. At triglyceride levels > 4,5 mmol/L there was a big difference between the methods, probably because of falsely low results from calculated LDL-cholesterol.