Prostata-adenocarcinom är den vanligaste cancerformen hos män. Anti-PSA och anti-P501S är frekvent använda antikroppar för diagnostisering av prostata-adenocarcinom i vävnadsbiopsier, men kan ha svag eller negativ infärgning av antigenen. Antikroppen anti-NKX3.1 har visat sig ha en högre sensitivitet och starkare infärgning än anti-PSA och anti-P501S vid prostata-adenocarcinom. Det rekommenderas att använda anti-NKX3.1 tillsammans med anti-PSA och/eller anti-P501S för att diagnostisera metastaserande prostata-adenocarcinom. Studiens syfte var att utvärdera och implementera antikroppen anti-NKX3.1 för diagnostisering av metastaserande prostata-adenocarcinom. Positiva vävnadskontroller (prostatavävnad och testisvävnad) och negativ vävnadskontroll (appendix) färgades med anti-NKX3.1 i Roche Ventana Benchmark ULTRA med immunohistokemisk färgning. Förbehandlingarna CC1 mild, CC1 standard och Proteas 1 samt visualiseringskiten OptiView samt UltraView jämfördes. Spädningen optimerades från 1:50-1:200. Vävnadssnitten graderades sedan från 1 (ingen infärgning)-6 (starkast infärgning). Visualiseringskitet OptiView samt förbehandlingen CC1 standard gav den starkaste infärgningen med anti-NKX3.1 och valdes som det optimala visualiseringskitet respektive förbehandlingen. Med tanke på materialkostnad och smidighet valdes spädningen 1:100 som den optimala spädningen. / Prostate adenocarcinoma is the most common form of cancer in men. Anti-PSA and anti-P501S are frequently used antibodies for diagnosing of prostate adenocarcinoma in tissue biopsies but may show weak or negative staining of the antigens. Antibody anti-NKX3.1 has been shown to have a higher sensitivity and stronger staining then anti-PSA and anti-P501S in prostate adenocarcinoma. It is recommended to use anti-NKX3.1 along with anti-PSA and/or anti-P501S to diagnose metastases of prostate adenocarcinoma. The purpose of the study was to evaluate and implement the antibody anti-NKX3.1 for diagnosis of metastatic prostate adenocarcinoma. Positive tissue controls (prostate and testis) and negative tissue control (appendix) were stained with anti-NKX3.1 immunohistochemical staining in Roche Ventana Benchmark ULTRA machine. The pre-treatments CC1 mild, CC1 standard and Protease 1 and the visualization kits OptiView and UltraView were compared. The dilution was optimized from 1:50-1:200. Immunohistochemical staining of the tissue was graded from 1 (no staining) - 6 (strongest staining). Visualization kit OptiView and pre-treatment CC1 standard gave the strongest staining with anti-NKX3.1 and were chosen as the optimal visualization kit and pre-treatment. Considering the material cost and flexibility, the 1:100 dilution was chosen as the optimal dilution.

Overweight (BMI 25–29) or obese (BMI over 30) pregnant women have an increased risk of complications during pregnancy and childbirth that can harm both women and their children.Due to increased risk for several complications such as of giving birth to large children, malformations or still births, the children have higher risk of obesity, blood pressure diseases and diabetes in childhood and later in life compared with children of normal weight. Various factors affect nutrient supply to the fetus such as activity of transport proteins, maternal and fetal blood flow to and from the placenta and placental metabolism. X-box-binding protein1 (XBP-1) is an important transcription factor that is part of the endoplasmic reticulum (ER)and facilitates the breakdown of misfolded proteins and gives ER good quality control over proteins. The purpose of this work was to study the XBP-1 protein in the placenta and see if the cells are stressed in women with obesity and compare levels of this protein with women who are normal weight. Placental expression of XBP-1 was analyzed by use of two different methods western blot including 36 women in three different groups, 12 women who were normal weight, 12 women who were obese and 12 women who were overweight. The second method was real- time polymerase chain reaction (RT- PCR) including 20 women in two different groups, 10 women who were normal weight and 10 women who were obese. The result showed that there was no protein detected in the placenta but there were bands in positive control. XBP-1 mRNA expression did not differ between study groups but there was a tendency towards a significant correlation mRNA and birth weight of the children. This indicate that it may a correlation between them, but our results should be confirmed with larger studies.

Background: Glomerular filtration rate (GFR) is a clinically important measurement of kidney function and estimating the GFR is of great importance in healthcare. Methods available today either lack in precision or are overly time consuming. Proenkephalin (PENK) has been shown to correlate well with the GFR and has therefore been proposed as a novel biomarker for kidney function. Aim: To evaluate a new point-of-care test for measuring PENK in blood and to assess its correlation to GFR measured by iohexol plasma clearance (mGFRiohexol). Materials and methods: Blood was collected from 21 patients with varying indications for the iohexol plasma clearance test. PENK was measured with IB10 Sphingotest penKid in whole blood, plasma and serum respectively. The concentration was correlated to the mGFRiohexol and results were compared to those from measurement of the routine markers for kidney function, i.e. creatinine and cystatin C. Results: Fourteen men and seven women were included. The median age was 57 years. PENK in plasma correlated weakly with mGFRiohexol (R2=0.22, p=0.042). No significant correlation was shown for PENK in whole blood or serum. Creatinine also showed a weak correlation with mGFRiohexol (R2=0.35, p=0.0046). In contrast, cystatin C was strongly correlated with mGFRiohexol (R2= 0.87, p<0.0001). Conclusion: Our results indicate that PENK is a biomarker of low clinical value for estimating the GFR. However, further studies are needed before this can be assured. Cystatin C, on the contrary, seems to be an accurate biomarker for estimating the GFR.

Respiratory tract infections are caused by different types of virus and bacteria and have become a global health issue. The detection and analysis of respiratory tract pathogens is nowadays performed commonly using PCR technology because of its high sensitivity. The aim of this study was to compare the performance of a new instrument, Starlet IVD with Maglead 12gC for extraction of nucleic acid from respiratory tract samples prior to real time PCR analysis. Frozen positive samples and fresh negative samples from patients were analyzed for respiratory viruses and respiratory bacteria. The samples were extracted on both Starlet IVD and Maglead 12gC. Real time PCR analysis was performed after the extraction to compare the performance between both instruments. A total of 44 virus samples were analyzed with Allplex respiratory panel 2 and 61 bacteria samples analyzed with Allplex respiratory panel 4. The result has shown an acceptable agreement of the PCR analysis (> 95%) for both panels between instruments. Starlet IVD has a higher Ct value compared to Maglead 12gC which led to a variation of 7,1% ± 3,6% in Ct value for Respiratory panel 2 and 10,1 % ± 5,3 % for Respiratory panel 4.  In conclusion, Starlet IVD is an automated instrument that will reduce hand-on-time, the risk of errors and improve the work routine for a better quality of results. Regardless the poor Ct-value, the new instrument has been able to detect almost all respiratory pathogens in the samples.

Bakgrund och syfte Procalcitonin (PCT) används i sjukvården vid uppföljning av patienter med bakterieinfektion. Under rådande corona-pandemi har studier visat att förhöjt PCT är associerat med utvecklingen av en svår form av Sars-Cov2-infektion. Genom att mäta PCT kan tecken till sekundär bakterieinfektion identifieras och utvecklingen av svår sjukdomsbild förutspås.På Universitetssjukhuset Örebro (USÖ) sker analysen av PCT på instrumentet Cobas e411. Under den pågående pandemin pågår det nu diskussion kring att överföra PCT-analysen till instrumenten ADVIA Centaur XPT på USÖ istället eftersom detta instrument även finns i Lindesberg och Karlskoga. Analysen skulle därmed kunna ske på alla tre laboratorier. Metod Venöst blod med olika koncentrationer av PCT samlades in från 21 patienter. Proverna användes till patientjämförelse. Till inomserieprecision användes patientprover som poolats till en hög och en låg nivå. Till totalprecisionen användes hög och låg kontroll. Datan från patientjämförelsen bearbetades i Excel och presenterades i linjära regressioner och Bland altman-plot. Som sensitivitetsanalys upprepades den statistiska bearbetningen med de sju högsta värdena exkluderade. Resultat och slutsats Patientjämförelsen mellan Cobas e411 och ADVIA Centaur XPT 1070 samt mellan ADVIA Centaur XPT 1070 och 1071 visade en bra korrelation. ADVIA Centaur XPT mäter dock lägre än Cobas e411, vilket ses i Bland altman-plotten som en metodbias på 30%. Detta ses även efter sensitivitetsanalys med högre värden exkluderade och datan bearbetats på nytt. Precisionsmätningarna visade bra precision med låga CV% för både ADVIA Centaur XPT 1070 och 1071.Med hänsyn till resultatet betraktas bytet till ADVIA Centaur XPT som en risk, varför beslutet fattas att fortsättningsvis analysera PCT på Cobas e411. / Background and purpose The PCT-value is used for observing the development of illness in patients with bacterial infection. During the Covid 19-pandemic, studies have shown that an increased PCT predicts a severe Covid-19.At the University hospital of Örebro, USÖ, the PCT assay is analyzed on the Cobas e411 instrument. The hospitals of Lindesberg and Karlskoga cannot perform the analysis due to a lack of this instrumentation.Since Centaur instruments are available at all three hospitals, performing the analysis on these instruments instead of the Cobas would enable all three hospital laboratories in the county to offer PCT analysis. Method Venous blood with different PCT values where collected from 21 patients and analysed with Siemens ADVIA Centaur XPT and Roche Cobas e411. Within-run precision was tested using pooled samples of low and high PCT concentrations. Between-run precision was tested using controls of low and high PCT concentrations. The data was examined by linear regression and Bland altman-plots using Excel. As a sensitivity analysis, the seven highest values were excluded, and the statistical analysis was repeated. Results and conclusion Despite the values correlating well in the linear regression and the assay had a high precision with a low CV% for both ADVIA Centaur XPT 1070 and 1071, the ADVIA Centaur XPT-instrument systematically measures 30% lower compared to Cobas e411. This method bias could lead to a misclassification of Covid 19-patients. With respect to our results, we consider analysis on ADVIA Centaur XPT to be risky and thus choose to continue analysing on Cobas e411.

Interferenser i klinisk kemi uppstår när en substans stör mätningen av olika komponenter i urin, cerebrospinalvätska och blod, vilket påverkar mätresultatet. Exempel på interferenser inkluderar hemolys, lipemi och ikterus. Hemolys kan orsakas både in vivo och in vitro, men in vitro är betydligt vanligare. Faktorer som påverkar in vitro hemolys inkluderar rörpost, provtagningsteknik och olika sjukdomar. Rörposttransport används för att transportera olika material, inklusive prov, till olika avdelningar på sjukhuset. Hemolys kan påverka analysen av vissa analyter som normalt förekommer i hög koncentration intracellulärt, exempelvis aspartataminotransferas (ASAT), laktatdehydrogena (LD) och kalium.   Syftet med denna studie var att undersöka om rörtransportsystemet på Universitetssjukhuset Örebro (USÖ) påverkar analysresultatet av kaliumprover och om förekomsten av hemolys ökade i kaliumprover som transporterades med rörpost. Studien utfördes med hjälp av direkt och indirekt potentiometrisk jonselektiv elektrodmetod (ISE) för att analysera nivåerna av kalium i plasma, samt spektrofotometri för att kvantifiera graden av hemolys i 129 prover som insamlades på akutmottagningen vid Universitetssjukhuset Örebro (USÖ). Ingen kliniskt relevant skillnad i hemolys eller kaliumkoncentration kunde påvisas i prover som transporterats med rörpost i jämförelse med manuell transport till laboratoriet. Rörposttransport är en kostnadseffektiv och snabb transportmetod och för de skillnader som observerades mellan transportmetoderna i denna studie bör andra faktorer som högre gravitationskraft och olika sjukdomstillstånd också beaktas. / Interference in clinical chemistry refers to a substance that disrupts the measurement of various components in urine, cerebrospinal fluid, and blood, affecting the obtained results. Hemolysis, lipemia, and icterus are examples of interferences. Hemolysis is caused by both in vivo and in vitro factors, but in vitro hemolysis is more common. Hemolysis affects analytes such as potassium, aspartaminotransferas (ASAT) and lactate dehydrogenase (LD). This study aimed to investigate whether the tube transport system at USÖ affects the analysis results of potassium.   129 samples were collected and each sample was divided into two portions a and b, sample a was then transported by tube and b manually. Samples were analyzed for hemolysis, potassium, and sodium using ion-selective electrode (ISE) methods. No clinically relevant difference in hemolysis or potassium concentration could be demonstrated in samples transported by pneumatic tube compared to manual transport to the laboratory. The use of tube transport is cost-effective and rapid despite the small difference observed, it should still be used.   In conclusion, while hemolysis can affect analytes, the observed difference between tube and manual transport in this study had no clinical significance. Other factors such as different G- forces and disease states may have played a role in explaining divergent results.

Bakgrund Assisterad befruktning är en vanlig behandling för infertilitet, vilket innebär oförmågan att bli gravid. Vid assisterad befruktning övervakas embryoutvecklingen med hjälp av Time Lapse Imaging-teknologi i en EmbryoScope och implantationsförmågan bedöms. Tidigare forskning har identifierat nukleära avvikelser (NE) i tidig embryoutveckling som kan påverka graviditetsfrekvensen (Pregnancy Rate, PR) och frekvensen av levande födda barn (live birth rate, LBR). Idag beräknas NE till fyra fenotyper (NEP): binukleation, mikronukleation, multinukleation samt splittrad nukleation.   Syfte Syftet med denna retrospektiva registerstudie var att undersöka sambandet mellan NE under tidig embryoutveckling och PR, LBR samt missfall för att förbättra embryoselektionen vid Reproduktionsmedicinskt centrum (RMC) på Örebro universitetssjukhus (USÖ).   Resultat Resultatet visade att 37,5% av de återförda blastocysterna i studien hade NE, och att PR och LBR påverkades av vilken NEP embryot hade. Embryon med binukleäritet hade högre PR och LBR än embryon med övriga kärnstatusar. KIDScore D5 visade sig vara ett bra selektionsverktyg för blastocyster återförda dag 5, men närvaron/frånvaron av NE påverkade kopplingen mellan KIDScore D5 och graviditet/barn på ett oväntat sätt, beroende på NEP.   Slutsats Embryoselektionen vid RMC på USÖ kan förbättras genom att ta hänsyn till kärnstatusen i tidig embryoutveckling, men en större studiepopulation behövs för att bekräfta korrelationen mellan alla variabler. / Background Assisted reproduction is a common treatment for infertility, which is the inability to become pregnant. In assisted reproduction, the development of embryos is monitored using Time Lapse Imaging technology in an EmbryoScope, and the implantation potential is assessed. Previous research has identified nuclear errors (NE) in early embryonic development that can affect pregnancy rates (PR) and live birth rates (LBR). NE is currently divided into four phenotypes (NEP): binucleation, micronucleation, multinucleation, and fragmented nucleation.   Objective The purpose of this retrospective registry study was to investigate the correlation between NE in early embryonic development and PR, LBR, and miscarriage to improve embryo selection at the Reproductive Medicine Center (RMC) at Örebro University Hospital (USÖ).   Results The results showed that 37.5% of the returned blastocysts in the study had NE, and that PR and LBR were affected by the NEP of the embryo. Embryos with binucleation had higher PR and LBR than embryos with other nuclear statuses. KIDScore D5 was found to be a good selection tool for blastocysts returned on day 5, but the presence/absence of NE affected the correlation between KIDScore D5 and pregnancy/childbirth in an unexpected way, depending on the NEP.   Conclusion Embryo selection at RMC at USÖ can be improved by considering the nuclear status in early embryonic development, but a larger study population is needed to confirm the correlation between all variables.

Syftet med projektet var att utvärdera hållbarheten av analyterna follikelstimulerande hormon (FSH), luteiniserande hormon (LH), progesteron (PRGE) och sexualhormonbindande globulin (SHBG) i primärrör förvarade i kyl, i serum såväl som plasma, på ADVIA Centaur XPT (Siemens Healthineers). Samtliga analyter analyserades i serum respektive plasma vid tidpunkterna 0, 24, 48, 72, 96 samt 168 h efter insamling. Analyspåverkan av förvaringstid i kyl och korrelationen mellan analyser i serum och plasma utvärderades. FSH, LH och SHBG uppvisade acceptabla medelbias vid samtliga tidpunkter i båda matriserna. PRGE uppvisade låg medelbias, <15 %, vid samtliga tidpunkter i serum men uppvisade medelbias >15 % i plasma. Analys i plasma skiljde sig minimalt från serum för FSH, LH och SHBG. För PRGE var skillnaderna lite större. Utifrån resultatet bedömdes FSH, LH och SHBG vara hållbara i en vecka i kyl (168 h) i både serum och plasma. PRGE bedömdes vara hållbart i kyl i en vecka i serum men inte godtagbar i plasma. Laboratoriets riktlinjer kan utvidgas för FSH, LH och SHBG att också genomföras i plasma. PRGE fortsatt bör genomföras i serum. / The purpose of the project was to evaluate the stability of the analytes follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (PRGE) and sex hormone-binding globulin (SHBG) in primary collection tubes stored refrigerated, in serum and plasma, on ADVIA Centaur XPT (Siemens Healthineers). All analytes were analyzed in serum and plasma at 0, 24, 48, 72, 96 and 168 h after collection. The impact of storage time on the analysis results and the correlation between analyzes in serum and plasma were evaluated. FSH, LH and SHBG obtained a mean bias <15 %, the threshold, at every moment in both matrices. PRGE showed a mean bias <15 % at every moment in serum but showed a mean bias >15 % in plasma. Analysis in plasma differed minimally from serum for FSH, LH and SHBG. For PRGE, the differences were slightly larger. This study considered FSH, LH and SHBG to be stable for one week (168 h) in both serum and plasma stored refrigerated. PRGE was considered to be stable for one week in serum stored refrigerated, and not acceptable in plasma. Analyzes of FSH, LH and SHBG can be carried out in plasma, while analyzes of PRGE should continue to be performed in serum.

Introduktion: Det perifera nervsystemet består av det autonoma nervsystemet och de perifera nervtrådarna som förmedlar afferenta och efferenta impulser mellan det centrala nervsystemet och perifera delen av kroppen. Från plexus brachialis förgrenar nervus ulnaris sig och fortsätter längst armen till handens ulnara del. Den vanligaste perifera nervskadan i övre extremiteter är ulnarisnervskada. Syftet: Syftet med studien var att undersöka om det förekommer någon signifikant skillnad i motorisk ledningshastighet (MCV) mellan underarm och över armbåge på nervus ulnaris bilateralt. Även att jämföra MCV och proximal latenstid mellan höger och vänster underarm och över armbåge. Metod: För att besvara syftet utfördes en tvärsnittsstudie på 31 friska deltagare i åldrarna 20– 40 år. Nervus ulnaris undersöktes med elektroneurografi bilateralt. Resultat: Resultatet visade att det förekommer en signifikant skillnad i MCV mellan underarm och över armbågen bilateralt på nervus ulnaris. Det påvisades en signifikant skillnad i MCV mellan höger och vänster sida över armbågen, men inte på underarm. Ingen signifikant skillnad förekommer i proximala latenstiden mellan höger och vänster underarm och över armbågen. Slutsats: De signifikanta skillnaderna som erhölls i MCV mellan underarm och över armbåge samt mellan höger och vänster sida över armbågen kan bero på stimuleringstekniken. Såsom armspositionen vid stimuleringen och att överarmar är generellt svårare att undersöka. Detta medför större risk för felkällor som kan påverka resultatet. / Introduction: The peripheral nervous system consists of the autonomic nervous system and the peripheral nerve fibers that mediate afferent and efferent impulses between the central nervous system and the peripheral part of the body. From the brachial plexus, the ulnar nerve branches and continues along the arm to the ulnar part of the hand. The most common and largest peripheral nerve injury in the upper extremities is ulnar nerve injury. Aim: The aim of the study was to investigate whether there is any significant difference in motor conduction velocity (MCV) between the forearm and above the elbow on the ulnar nerve bilaterally. Also, to compare MCV and proximal latency between right and left forearm and above elbow. Method: To answer the purpose, a cross-sectional study was performed on 30 healthy participants aged 20–40 years. The ulnar nerve was examined with electroneurography bilaterally. Results: The result showed that there is a significant difference in MCV between the forearm and above the elbow bilaterally on the ulnar nerve. A significant difference in MCV was demonstrated between the right and left sides over the elbow, but not on the forearm. There was no significant difference in the proximal latency between the right and left forearm and above the elbow. Conclusion: The significant differences obtained in MCV between forearm and above elbow and between right and left side above elbow may be due to the simulation technique. Such as the arm position during the stimulation and that upper arms are generally more difficult to examine. This entails a greater risk of error sources that can affect the result.

Background: Mesenchymal stromal cells (MSCs) are multipotent cells that can differentiate into various cell types and possess immunomodulatory and anti-inflammatory effects, making them interesting candidates for therapeutic applications. MSCs are present in small quantities in tissues like bone marrow and therefore need to be expanded while preserving their essential characteristics. They should adhere to plastic, differentiate into osteocytes, adipocytes and chondrocytes and express specific cell surface markers. Currently, the “golden standard” culture media supplement is fetal bovine serum (FBS). However, there is a potential contamination risk of MSCs by xenogeneic and zoonotic infectious agents, which can trigger an immune response. As an alternative, xeno-free serum supplements derived from human sources, e.g., human serum (HS) can be used.  Aim: This study aimed to identify and characterize human bone marrow derived MSCs and examine the effects different supplements have on the cells.  Methods: MSCs were cultured in 10% FBS, 2% FBS and 10% HS for 20 -21 days. Differentiation was induced and the potential was detected with immunocytochemistry. Cell surface markers CD73, CD90, CD105 and CD45 were identified with flow cytometry.  Results and Conclusion: There was no significant difference in morphology, differential potential or immunophenotype between the different serum conditions. However, HS-supplemented culture media resulted in a significantly higher number of cells with 1 x 107 cells after 20 days without affecting their differentiation potential and immunophenotype in comparison to 10% FBS with 2.2 x 106 cells (p=0.0004). MSCs cultured in 2% FBS resulted in the least number of cells (9.9 x 105) after 21 days of expansion.