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The tetracycline repressor (tetR)-regulated system is a widely used tool to specifically control gene expression in mammalian cells. Based on this system, we generated a human osteosarcoma cell line, which allows for the inducible expression of an EGFP fusion of the TAR DNA-binding protein 43 (TDP-43), which has been linked to neurodegenerative diseases. Consistent with previous findings, TDP-43 overexpression led to the accumulation of aggregates and limited the viability of U2OS. Using this inducible system, we conducted a chemical screen with a library that included FDA-approved drugs. While the primary screen identified several compounds that prevented TDP-43 toxicity, further experiments revealed that these chemicals abrogated the doxycycline-dependent TDP-43 expression. This antagonistic effect was observed with both doxycycline and tetracycline, and in several Tet-On cell lines expressing different genes, confirming the general effect of these compounds as inhibitors of the tetR system. Using the same cell line, a genome-wide CRISPR/Cas9 screen identified epigenetic regulators such as the G9a methyltransferase and TRIM28 as potential modifiers of TDP-43 toxicity. Yet again, further experiments revealed that G9a inhibition or TRIM28 loss prevented doxycycline-dependent expression of TDP-43. In summary, we have identified new chemical and genetic regulators of the tetR system, thereby raising awareness of the limitations of this approach to conduct chemical or genetic screening in mammalian cells.

Cancer cell migration is a driving mechanism of invasion in solid malignant tumors. Anti-migratory treatments provide an alternative approach for managing disease progression. However, we currently lack scalable screening methods for identifying novel anti-migratory drugs. To this end, we develop a method that can estimate cell motility from single end-point images in vitro by estimating differences in the spatial distribution of cells and inferring proliferation and diffusion parameters using agent-based modeling and approximate Bayesian computation. To test the power of our method, we use it to investigate drug responses in a collection of 41 patient-derived glioblastoma cell cultures, identifying migration-associated pathways and drugs with potent anti-migratory effects. We validate our method and result in both in silico and in vitro using time-lapse imaging. Our proposed method applies to standard drug screen experiments, with no change needed, and emerges as a scalable approach to screen for anti-migratory drugs. The spatial positioning of cultured glioblastoma cells is used to estimate cell motility and drug effects from single end-point images in vitro.

En ökad utbredning av vankomycinresistenta enterokocker (VRE) har setts i Sverige sedan 2007. Bakteriemi orsakad av VRE är mycket svårbehandlad, varför snabbare tillförlitlig resistensdiagnostik är betydelsefullt för att minska dödlighet, vårdtider, vårdkostnader och belastning på sjukvårdssystemet. På mikrobiologilaboratoriet, Region Jönköpings län (RJL), tar idag identifiering av fenotypisk vankomycinresistens vid optimala förhållanden 6 timmar, räknat från att enterokocker konstaterats växa i blodet. Resistensgenerna vanA och vanB, som bland andra orsakar vankomycinresistens hos enterokocker, kan genetiskt verifieras med loop-mediated isothermal amplification men tar idag upp till ett dygn då bakteriekolonier används som analysmaterial i arbetsrutinen på molekylärbiologilaboratoriet, RJL. Syftet med studien var att utvärdera bakteriepellet som analysmaterial för genetisk identifiering av vanA och vanB, på Genie® II Mk2 med eazyplex® VRE basic, hos enterokocker från positiva blododlingar. För att utvärdera bakteriepellet som analysmaterial analyserades isolat av Enterococcus faecium (n=17) och Enterococcus faecalis (n=5) från bakteriepellets tillverkade från simulerade positiva blododlingar med eazyplex® VRE basic på Genie® II Mk2, varpå resultaten jämfördes mot isolatens faktiska närvaro/frånvaro av vanA/vanB. Samstämmigheten av de uppmätta- och de förväntade resultaten var fullständig, vilket indikerar att bakteriepellet med hög tillförlitlighet kan användas som analysmaterial till eazyplex® VRE basic för att påvisa vanA och vanB hos enterokocker i blododlingar. An increased prevalence of vancomycin-resistant enterococci (VRE) has been observed in Sweden since 2007. Treating bacteremia caused by VRE is difficult, which is why faster, and reliable resistance diagnostics are important. At the Microbiology laboratory, Region Jönköping County, the identification of phenotypic vancomycin resistance under optimal conditions takes 6 hours from when growth of enterococci in blood is determined. The genes vanA and vanB, which among others cause vancomycin resistance, can be genetically verified by loop-mediated isothermal amplification, but takes up to one day since bacterial colonies are used as analysis material. The aim of the study was to evaluate bacterial pellet as an analytical material for genetic identification of vanA and vanB, on Genie® II Mk2 with eazyplex® VRE basic, in enterococci from positive blood cultures. To evaluate the bacterial pellet, isolates of Enterococcus faecium (n=17) and Enterococcus faecalis (n=5) from bacterial pellets made from simulated positive blood cultures were analyzed with eazyplex® VRE basic on the Genie® II Mk2, and the results were compared to the actual presence/absence of vanA/vanB in the isolates. The complete coherence between the expected and measured results indicates that the bacterial pellet can be used as an analytical material for eazyplex® VRE basic.

Background: Vancomycin-resistant enterococci (VRE) is a common nosocomial infection. It classifies as Enterococcus faecalis or faecium carrying vanA or vanB gene that alters the bacterial cell wall hence lowering affinity for Vancomycin. Screening for VRE in Swedish hospitals are performed with stool sample pre-grown in selective broth followed by PCR and culture on selective media. Viasures Vancomycin resistance, Real Time PCR Detection Kit indicates that pre-growth in broth is not needed for the analyze. Aim: Comparison between the PCR kit and the subsequent culture on chromogenic agar with or without pre-growth in selective broth. Method: E. faecium with vanA gen (CCUG 36804) and E. faecalis with vanB gen (ATCC 51299) were suspended in different concentrations and added to ESwab transport medium. Thereafter small samples from the ESwab tube were enriched in selective broth. Samples from both selective broth and ESwab medium were analyzed with Viasures PCR kit on BD-MAX system and cultivated on chromogenic agar. Results: With pre-growth in selective broth the genes were found in every sample regardless of pre-concentration in the ESwab medium. Without enrichment the PCR kit always amplified the genes when the concentration was 40 000 cfu/ml for E. faecium (vanA) and ≥ 10 000 cfu/ml for E. faecalis (vanB). Colonies grew on chromogenic agar in every concentration from both ESwab and selective broth. Conclusion: Culture on chromogenic agar is comparable with or without pre-growth in selective broth but Viasure’s PCR kit is not equal for both methods in lower concentrations of the bacteria.

Giardia is a parasite that causes disease in both humans and animals all around the world. Approximately 200 million people are infected each year. The parasite is spread mainly through contact with contaminated feces but can also be spread through food and water contaminated with infectious cysts. Symptoms that may occur during infection could be diarrhea and abdominal pain. Not all host organisms get symptoms and there are lots of asymptomatic carriers. The aim of this report was to compare and evaluate the performance of three different methods used to detect Giardia in dogs and cats. Today the gold standard is detecting cysts with immunofluorescence, which was one of the methods evaluated. One cyst visible was enough to say the test was positive. The other two methods were an ELISA and a rapid test, and both detected soluble Giardia antigen. For both methods, color development occurred in case of positive results. The ELISA wells were read spectrophotometrically at 450 nm. During this project, 294 tests from randomly chosen dogs and cats were analyzed. Of all these tests, 103 were analyzed with all three methods, whereas 191 tests were only analyzed with immunofluorescence and ELISA. When analyzing the results, the number of positive test results was very similar between the three methods and there was no statistically significant difference between them. Therefore, when price, time and results were taken into consideration, the method recommended was ELISA.

Multiple myeloma, or myeloma, is a bone marrow cancer which characterizes by uncontrolled proliferation of mutant plasma cells. It is a disease that claims many lives every year, mostly due to the absence of curative treatment. Finding a suitable treatment is therefor of great importance. One way to study different diseases is to use a gene editing method for knockdown or knockout of specific genes. The main aim of this project was to design guide RNAs, to be able to use CRISPR/Cas9 for knockout of the two genes BMPR1A and BMPR2 in different myeloma cell lines (KJON, INA-6 and IH-1). This, to be able to study the expression and function of these genes. Further aim of the project was to investigate potential SMAD activation by treatment with different bone morphogenetic proteins (BMPs). However, due to limited time this could not be carried through. Six guide RNAs were designed and ligated into pLentiCRISPRv2. Plasmid amplification was done by transformation of Escherichia coli. To check the quality of the plasmids, PCR, gel electrophoresis and Sanger sequencing was performed. The results from the gel electrophoresis showed that nine of the twelve samples for BMPR1A and seven of the thirteen samples for BMPR2, that were tested, were positive. The results from the Sanger sequencing confirmed that all guides that were tested (BMPR1A 3.2.3, BMPR1A 4.2.2, BMPR2 1.1.4 and BMPR2 2.1.2), were properly ligated into the plasmids. The main aim of the project was successfully accomplished, but additional work is needed for any further conclusions.

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.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) är en viktig metod för artidentifiering av mikroorganismer på kliniska mikrobiologiska laboratorier. Enligt rekommendationer från Bruker Daltonics skall matrixapplicering ske relativt omedelbart efter att mikroorganismer har applicerats. Syftet med studien var att utvärdera effekter på artidentifiering vid förlängda tidsintervall mellan applicering av mikroorganismer och matrix med MALDI-TOF MS. Målet var att optimera arbetsflödet på mikrobiologilaboratoriet, laboratoriemedicin, Region Jönköping län (RJL). I studien undersöktes hur identifieringen av mikroorganismer påverkades av applicering avbakterie- och jästsvampstammar (n = 267) och α-Cyano-4-hydroxycinnaminsyra (HCCA)-matrix på en MALDI-provplatta vid varierande tidsintervall upp till tre timmar. Analys med MALDI-TOF MS genomfördes med masspektrometrarna MALDI Biotyper Sirius IVD System och Bruker MicroFlex LT. Resultaten visade att förlängda tidsintervall mellan applicering av mikroorganismer och matrix upp till tre timmar inte hade någon signifikant effekt på scorevärden vid artidentifiering, vilket innebar att tidsintervallen är acceptabla för artidentifiering med MALDI-TOF MS. Detta ger värdefull information för att optimera arbetsflödet och öka effektiviteten på mikrobiologilaboratoriet. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an important method for species level identification in clinical microbiology laboratories. According to recommendations from Bruker Daltonics matrix application should occur relatively immediately after microorganisms have been applied. The aim of the study was to evaluate the effects on microbial identification at extended time intervals between microbial and matrix application using MALDI-TOF MS. The objective was to optimize the workflow at the microbiology laboratory, laboratory medicine, Region Jönköping län (RJL). The study investigated the impact on identification of microorganisms between the application of bacterial and yeast strains (n = 267) and α-Cyano-4-hydroxycinnamic acid (HCCA) matrix on a MALDI target plate of various time intervals up to three hours. MALDI-TOF MS was performed using MALDI Biotyper Sirius IVD System and Bruker MicroFlex LT mass spectrometers.The results showed that a delay of matrix application up to three hours had no significant effect on score values, indicating that time did not affect the accuracy of microbial identification using MALDI-TOF MS. Consequently, the study provided valuable information for optimizing the workflow and increasing the efficiency of the microbiology laboratory.

Neisseria meningitidis är en bakterie som kan leda till invasiv sjukdom eller endast orsaka bärarskap i nasopharynx. Bakterien delas in i olika serogrupper och klonala komplex. Vissa av dessa grupper och klonala komplex förekommer endast hos invasiva isolat och andra bland bärare, vilket tyder på att det finns genetiska skillnader mellan invasiva och bärarisolat. I denna studie undersöktes genen pilE som kodar för PilE proteinet och ingår i bakteriens pili. Proteinet finns i två klasser, klass 1 och klass 2. Metoden som användes för att studera eventuella skillnader i förekomst och uttryck av pilE genen var digital droplet PCR (ddPCR). Både DNA och RNA kvantifierades med ddPCR för att undersöka antalet kopior av pilE genen (DNA) samt dess uttryck (RNA) mellan invasiva isolat och bärarisolat, mellan klass 1 pilE isolat och klass 2 samt fördelning av klasserna i isolattyperna. Principen för ddPCR är att dela ett prov till tiotusentals nanodroppar där individuella droppar genomgår PCR för en senare avläsning av flourescens från prober. Resultatet visade skillnad mellan bärarisolat och invasiva isolat där de invasiva isolaten visade mindre uttryck av pilE än bärarisolat. Klass 2 isolat hade signifikant färre genuttryck än klass 1 isolat och invasiva isolat visade klass 2 i högre utsträckning än bärarisolat., Neisseria meningitidis is a bacterium that can cause invasive disease or carriage in the nasopharynx. The bacteria are divided into different serogroups and clonal complexes. Specific serogroups and clonal complexes are more frequent or are only found among invasive isolates or in carriage isolates. This study has investigated pilE, a gene that encodes the PilE protein located in the bacteria´s pilin and the protein is found in either class 1 or class 2. digital droplet PCR was used to investigate differences in presence and expression of the gene pilE. Both DNA and RNA was quantified to study the difference in copy number of the pilE gene (DNA) and its expression (RNA) between invasive isolates and carriage isolates but also between class 1 isolates and class 2 isolates. The distribution of classes between the isolate types was also investigated. The ddPCR method divides a sample into thousands of nanodroplets and a PCR reaction occurs in each droplet followed by droplet reading where fluoroscens from probes is measured. The difference that could be seen was that the invasive isolates expressed pilE in lower copies. Class 2 isolates had a significantly lower gene expression than class 1 isolates and invasive isolates expressed class 2 in a higher frequency.

Immunocytokemi på utstryk är en metod för sortering av celltyper och identifiering av maligna celler. Metoden anses som ett aktuellt alternativ till den redan etablerade immunocytokemi på cellblock-metoden vid diagnostik av maligniteter. Syftet med examensarbetet är metodutvärdering och etablering av immunocytokemi på utstryk för sortering och identifiering av olika celltyper och maligna celler, samt metodjämförelse mellan utstryk och cellblock. Fem prover från pleuravätska framställdes till utstryk och cellblock. Immunocytokemi med antikropparna anti-Epithelial Specific Antigen, anti-Cytokeratin 7 och anti-Carcinoembryonic Antigen tillämpades på dessa. Resultatbedömning omfattade bevarandet av morfologin, färgupptag och ospecifik färgning hos celler och bakgrund, samt möjligheten att ställa säker diagnos för utstryk. Resultat visar färgupptag och viss bevarad morfologi hos utstryk. Samtliga utstryk har ospecifik färgning och kontamination som medför svårighet att urskilja celltyper och identifiera maligna celler. Möjlighet att ställa diagnos med utstryk varierar med preparaten. Färgningen hos samtliga cellblock är robust och mer specifik för celler och bakgrund. Examensarbetet når slutsatsen att etablering av immunocytokemi- metoden på utstryk är möjligt och bör tillämpas i vissa situationer. Etablering som rutinmetod är olämpligt. Framställning av preparat med hög kvalitet och specifika markörer är utvecklingsområden för laboratoriet och vidare forskning., The method of immunocytochemistry on smears is utilized for categorizing cell types and identifying malignant cells. Immunocytochemistry on smears is a prospective alternative to the already implemented cell block method when diagnosing malignancies. The thesis aims to evaluate and implement immunocytochemistry on smears for categorizing and identifying cell types and malignant cells, along with comparing smears and cell blocks. Five pleural samples were processed into smears and cell blocks. Staining of these occurred with antibodies anti- Epithelial Specific Antigen, anti-Cytokeratin 7 and anti-Carcinoembryonic Antigen. Evaluated factors included preservation of morphology, stain uptake and non-specific staining for cells and background and finally the ability to generate a definitive diagnosis for smears. Results demonstrate stain uptake and preservation of morphology in some smears. All smears demonstrate non-specific staining and contamination, causing difficult categorization and identification. Generating a definitive diagnosis varies among samples. The thesis concludes that immunocytochemistry is a suitable platform for smears and a prospective implementation at the laboratory in certain situations. Immunocytochemistry on smears is not eligible as a routine method. Preparation of high-quality smears and specific antibody markers are deemed points of interest for the laboratory and future studies.

Background: Platelets are the smallest cells in the blood and are involved in the wound healing process. Platelet transfusion is an important part of treating illnesses such as thrombocytopenia and to prevent and stop major bleedings. Platelets for transfusion are received after processing of whole blood through automated or semi-automated systems and pooling of interim platelet units received after the processing. A limit is set for the minimum total estimated platelet yield in a pool to ensure that the final product consists of enough platelets. Aim: The aim of this study was to evaluate the platelet yield index received from Reveos automatic blood processing system by comparing it to the fluorescent flow cytometry method on Sysmex cell counter. Material and method: Whole blood from 147 donors were processed in the Reveos automated blood processing system into the separate components. All the interim platelet units received a platelet yield index from Reveos. A sample from each interim platelet unit was collected and analyzed on Sysmex in the channel for fluorescent flow cytometry. Result: The difference between the platelet count from the two methods was 45 ±26 x109/unit (p

Introduktion: Hyalint brosk består av kondrocyter inbäddade i ett extracellulärt matrix som täcker alla kroppens synoviala leder. I dag är skador på hyalint brosk ett folkhälsoproblem med begränsade behandlingsmöjligheter. Mesenkymala stamceller har i flertalet studier visat sig vara en lovande kandidat för framtida behandling av broskskador, eftersom dessa kan differentiera till broskproducerande kondrocyter. Modern behandling av broskskador innefattar odling av patientens egna kondrocyter eller stamceller för att sedan återföra dem till broskskadan. För att göra detta effektivt behöver optimering av odlingsförhållanden för aktuella celler utföras. Syfte: Att undersöka tillväxt hos kondrocyter samt mesenkymala stromaceller från benmärg i varierande odlingsförhållanden, detta med avseende att utvärdera vilka tillsatser som mest gynnar cellernas viabilitet och proliferation. Material & Metod: Framtagning av kondrocyter och mesenkymala stromaceller från benmärg i denna studie utfördes på överblivet ortopediskt material från en individ. Cellerna kollagenasbehandlades innan de odlades ut i ett standardmedium under tre veckor. Därefter odlades de i närvaro av olika tillsatser under sex dagar. Cellernas viabilitet mättes genom relativ fluorescens med CellTiter-Blue® reagens medan proliferationen mättes genom cellräkning med flödescytometri. Resultat: Det fanns en signifikant skillnad (ANOVA, p, Introduction: Articular cartilage consists of chondrocytes and covers all the synovial joints in the body. Articular cartilage injuries are today a public health issue with limited orthopedic treatments. Mesenchymal stem cells have shown to be a promising candidate for future treatment of articular cartilage injuries due to their many abilities. Modern treatment of cartilage damage involves cultivation of the patient´s own chondrocytes and stem cells. To make this treatment as effective as possible the culture conditions for chondrocytes and stem cells must be optimized. Aim: To study cell growth in chondrocytes and mesenchymal stromal cells from bone marrow in varying culture conditions, this regarding to evaluate the most optimal culture conditions for cell viability and proliferation. Method: Cell extraction in this study was performed on leftover orthopedic material. The cells were treated with collagenase and then cultured in a standard medium for three weeks before they were cultured in presence of various factors for six days. Cell viability was measured by relative fluorescence while proliferation was measured by cell counting. Result: There was a statistically significant difference (ANOVA, p

Introduktion: Blodtryck är en livsviktig funktion som bidrar till syretillförseln till kroppens olika organ. Blodtryck går att mäta och är en viktig indikation på det kardiovaskulära systemets funktion. Det finns olika metoder att mäta blodtryck, bland annat auskultatoriskt, automatiskt och med hjälp av Dopplerteknik. Syftet var att jämföra de tre olika metoderna för att se om det förelåg en signifikant skillnad mellan metoderna. Material och metod: Studien bestod av 30 deltagare. Studiedeltagaren vilade i tio minuter före första blodtrycksmätningen, därefter mättes deras blodtryck i en slumpad ordning med auskultatorisk, automatisk och med Dopplermetod. För att kontrollera om det förelåg en statistisk signifikant skillnad användes en variansanalys (ANOVA) och ett parat t-test. Resultat och slutats: Det förelåg ingen signifikant skillnad mellan de systoliska blodtrycken. Däremot mellan de diastoliska trycken registrerat med den auskultatoriska och automatiska metoden förelåg det en signifikant skillnad, men den anses inte ha en klinisk betydelse., Introduction: Blood pressure is a vital function that contributes to the supply of oxygen to the body’s various organs. Blood pressure can be measured and is an important indicator of the cardiovascular system function. There are different methods to measure blood pressure, these are, for example auscultatory, automatic and with the help of a Doppler technique. The aim was to compare the three different methods and see if there was a significant difference between them. Material and method: The study consisted of 30 participants. The participant rested for ten minutes before the first examination and then their blood pressure was measured in a random order by the three methods: auscultatory, automatic and the Doppler method. To process the collected data an analysis of variance test (ANOVA) and a paired t-test were used. Results and conclusion: There were no significant difference between the systolic blood pressures. In contrast, there was a difference between the diastolic blood pressures registered with the auscultatory and automatic methods, but this was not considered to be of clinical significance.

Pulmonary oedema and pulmonary hypertension are often associated with heart failure. The Balb/CJ and C57BL/6J mouse strains have been shown to exhibit distinct genetic susceptibilities to heart failure. Balb/CJ develops reduced heart function associated with pulmonary edema and pulmonary hypertension when treated with angiotensin II and a high-salt diet (ANG II + Salt) while C57BL/6J does not. The aim of this study was to investigate the effects of ANG II + Salt on lung damage by studying the potential immune cell infiltration in Balb/CJ mice and to determine whether differences in expression levels of angiotensin II type 1 and 2 (AT1 and AT2) receptors contribute to these effects. Protein expression of macrophages, neutrophils, AT1 receptors, and AT2 receptors in lung samples from six Balb/CJ mice and seven C57BL/6J mice treated with ANG II + Salt were analysed using Western blot. Five untreated mice from each strain were used as controls. The results indicated a significant difference (p=0.009) in the expression of macrophages between Balb/CJ mice treated with ANG II + Salt with a mean value of 17.30 and C57BL/6J mice treated with ANG II + Salt with a mean value of 9.34. No significant differences were observed in the expression of neutrophils, AT1 receptors, or AT2 receptors between the groups. Contrary to our expectations, the results did not support immune cell infiltration in Balb/CJ mice lungs or show higher AT1 receptor levels in Balb/CJ mice compared to C57BL/6J mice. Further research is required to understand the underlying mechanisms involved.

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.

Introduktion Genom dynamisk spirometri går det att beräkna kvoten mellan forcerad exspiratorisk volym under en sekund och forcerad vitalkapacitet (FEV1/FVC) där ett värde på ≤0,7 tyder på obstruktiv lungsjukdom. Mobil spirometriutrustning skulle kunna möjliggöra tidigare diagnostisering av obstruktiva lungsjukdomar men även fungera som ett kostnadseffektivt alternativ vid enklare undersökningar. Syftet med studien var att jämföra resultat erhållna vid dynamisk spirometri utförd på en stationär respektive en mobil spirometriutrustning. Material och metod I studien deltog totalt 36 försökspersoner. Data samlades in mellan februari-mars 2023 och bestod av FVC och FEV1 erhållna från dynamisk spirometri på Vyntus™ One (stationär spirometriutrustning) och Aiocare™ Vyaire (mobil spirometriutrustning). Ett parat t-test (signifikansnivå 0,05) utfördes för att avgöra om det förelåg någon signifikant systematisk skillnad för data mellan de båda utrustningarna och förekomsten av slumpfel illustrerades med Bland-Altman-diagram. Intraklasskorrelationskoefficient (ICC) beräknades för samtliga variabler. Resultat och slutsats En mycket god korrelation mellan utrustningarna förelåg med ett ICC>0.9 för samtliga variabler. Ingen systematisk skillnad kunde påvisas för uppmätt FVC, FEV1 eller kvoten FEV1/FVC mellan Vyntus™ One och Aiocare™ Vyaire (p >0,05) och slumpfelet bedömdes acceptabelt., Introduction Dynamic spirometry enables calculating the ratio of forced expiratory volume under one second and forced vital capacity (FEV1/FVC) where ≤0,7 suggests obstructive lung disease. Mobile spirometer equipment could assist in earlier detection of obstructive lung diseases, but also function as a cost-efficient alternative. The purpose of the study is to compare results obtained with dynamic spirometry performed on a stationary and a mobile spirometry equipment. Material and method A total of 36 subjects participated in the study. Data collected between February-March 2023 consisted of FVC and FEV1 obtained from dynamic spirometry on Vyntus™ One (stationary spirometry equipment) and Aiocare™ Vyaire (mobile spirometry equipment). A paired t-test (0,05 level of significance) determined if there was any significant systematic difference between data from the two types of equipment and the magnitude of sampling error was visualized by Bland-Altman plots. Intraclass correlation coefficient (ICC) was calculated for all variables. Result and conclusion Excellent correlation with ICC>0,9 for all variables. There was no significant difference for measured FVC, FEV1 or FEV1/FVC between Vyntus™ One and Aiocare™ Vyaire (p >0,05) and the sampling error was considered acceptable.

Tillväxthormon och insulinliknande tillväxtfaktor-1 är blodprover som tas vid misstanke om akromegali. Immulite 2000 XPi är ett instrument som använder chemiluminescent microparticle immunoassay metodik för att kvantitativt mäta koncentrationen av GH och IGF-1. Analys av biokemiska markörer påverkas av flera olika faktorer. En viktig sådan faktor är hemolys. Hemolys innebär att de röda blodkropparna går sönder, vilket leder till frisättning av dess innehåll såsom hemoglobin i serum/plasma. Syftet med detta projekt var att undersöka hur hemolys påverkar resultatet av GH- och IGF-1-prover på immulite 2000 XPi. Utifrån erhållna resultat kommer klinisk kemi vid Universitetssjukhuset Örebros laboratorie rutiner vid hemolytiska prover på GH eller IGF-1 att uppdateras . Hemolys tillverkades och tillsattes till olika serumprover med låg och hög nivå av GH respektive IGF-1 för att erhålla prover med varierande grad av hemolys. Resultatet visade att ökande hemolysindex korrelerar med GH- respektive IGF-1-koncentrationer, med undantag för den låga GH koncentrationen som inte uppvisade någon korrelation till hemolysindex. En 10%:ig skillnad av GH och IGF-1- koncentrationer uppnåddes vid en ökning av hemolysindex med 555,40 mg/dL för GH respektive 333,3 mg/dL för IGF-1.Utifrån resultaten var hemolysgränsen likvärdig med tillverkarens gränser. Därför kan klinisk kemi på Universitetssjukhuset Örebro fortsätta med nuvarande hemolysgränser., Growth hormone and insulin-like growth factor-1 are blood samples taken upon suspicion of acromegaly and for follow-up of acromegaly treatment. Immulite 2000 XPi is an instrument that uses chemiluminescent microparticle immunoassay method to quantitatively measure the concentration of GH and IGF-1. Analysis of biochemical markers is affected by several different factors. An important such factor is hemolysis. Hemolysis means that the red blood cells break, whose contents leak into the serum/plasma. The study aimed to investigate how hemolysis affects the results of GH and IGF-1 samples on the Immulite 2000 XPi. The results will determine the routines for hemolytic samples for GH or IGF-1 at Örebro University Hospital. Hemolysis was produced and added to various serum samples with low and high levels of GH and IGF-1 to obtain samples with varying degrees of hemolysis. The results showed that increasing hemolysis index correlates with GH and IGF-1 concentrations, with the exception of low GH concentration, which did not show any correlation to hemolysis index. A 10% difference in GH and IGF-1 concentrations was achieved with an increase in hemolysis index of 555.40 mg/dL for GH and 333.3 mg/dL for IGF-1. Based on the results, the hemolysis limit was equivalent to the manufacturer's limits. Therefore, clinical chemistry at Örebro University Hospital can continue with current hemolysis limits.

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 % 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 % 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.

Diabetes mellitus is one of the most common diseases worldwide. The continued prevalence increase, together with the complexity of the disease with severe long-term complications makes efficient diagnostical and monitorial methods very important. The determination of glycated hemoglobin, HbA1c, plays an important role in both, underlying the importance of high-quality analytical instruments. Whenever a new instrument is introduced into routine clinical use an evaluation is needed. This study evaluated the agreement between the new Tosoh G11 HbA1c high performance liquid chromatography system as a replacement for the Bio-Rad D-100 at the Chromatographic Department of the Institute of Laboratory Medicine, Faculty of Medicine, University of Debrecen. In total, 66 samples were analyzed using both analyzers and a Bland-Altman comparison was performed together with a correlation study. Additionally, a linearity check, optimal error and routine error testing was carried out specifically for the Tosoh G11. The results showed high agreement between measurements, r=0.9975 (p

Karpaltunnelsyndrom är den vanligaste formen av infångningsneuropati hos den vuxna befolkningen. Kända riskfaktorer för karpaltunnelsyndrom inkluderar arbetsskador som upprepad och repetitiv användning av handen och handleden, men även andra orsaker som graviditet, kön, eventuella frakturer med mera. Elektroneurografi (neurografi) är en klinisk undersökningsmetod som används för att diagnostisera och gradera nervskadan av karpaltunnelsyndrom. Syftet med studien var att jämföra motorisk och sensorisk nervledningshastighet och amplitud mellan arbetsrelaterade skador och andra orsaker vid bekräftad karpaltunnelsyndrom. Patienter rekryterades från avledningen för klinisk neurofysiologi på Gävle sjukhus som var kallade för en neurografiundersökning med frågeställning karpaltunnelsyndrom. Inklusionskriterier i studien var patienter, kvinnor och män i åldern 18–65 år med bekräftad karpaltunnelsyndrom. Neurografi utfördes motorisk och sensoriskt på n. medianus och n. ulnaris bilateralt. Vidare undersöktes n. medianus bilateralt med 14–7 metoden. Ingen signifikant skillnad hittades mellan arbetsskador och andra orsaker för karpaltunnelsyndrom vid jämförelse mellan motorisk och sensorisk nervledningshastighet samt amplitud. Ingen skillnad hittades i 14–7 metoden vid registrering av n. medianus mellan grupperna. Inget samband hittades mellan kroppslängd och motorisk nervledningshastighet., Carpal tunnel syndrome is the most common form of entrapment neuropathy in the adult population. Known risk factors for carpal tunnel syndrome include work related injuries such as repeated and repetitive use of hand and wrist, but also other causes such as pregnancy, gender, possible fractures and more. Electroneurography (neurography) is a clinical examination method used to diagnose and grade the nerve damage of carpal tunnel syndrome. The purpose of this study is to compare motor and sensory nerve conduction velocity and amplitude between work related injuries and other causes when confirmed carpal tunnel syndrome. Patients were recruited from the neurophysiology department at Gävle hospital who were called for a neurography examination with a question of carpal tunnel syndrome. Inclusion criteria in the study were patients, women, and men between the age 18-65 years with confirmed carpal tunnel syndrome. Neurography were performed bilaterally for both motor and sensory function of the median and ulnar nerve. The median nerve was also examined bilaterally with the 14-7 method. No difference was found between work related injuries and other causes in carpal tunnel syndrome when comparing motor and sensory nerve conduction velocity and amplitude. No difference was found in the 14-7 method when registering the n. medianus between the groups. No correlation was found between body height and motor nerve conduction velocity.

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.

Introduktion: Sepsis är utan behandling ett livshotande tillstånd, en korrekt behandling behöver en snabb och tillförlitlig artidentifiering. Olika prepareringsmetoder av positiva blododlingar inför direkt identifiering med MALDI-TOF finns. MBT Sepsityper® IVD Kit är en CE-märkt metod. För att följa IVDR-direktivet behöver Sepsityper jämföras med den nuvarande in-house-metoden. Dessa två metoder jämfördes utifrån förmåga att ge ett tillförlitligt, snabbt resultat där även den praktiska metoden samt ekonomiska aspekten undersöktes Material och metod: Alla positiva blododlingar som undersöktes preparerades med in- house-metoden och Sepsityper innan analys med MALDI-TOF MS. Med in-house-metoden användes saponin för att lysera de humana cellerna medans Sepsityper använde lyseringsbuffert. Resultatets tillförlitlighet presenteras som scorevärde där >2 är tillförlitligt till speciesnivå, 1,7–1,99 till genusnivå och, Background: Sepsis without treatment is a life-threatening condition, a rapid as well as reliable identification of the bacteria is needed. Different preparation methods of positive blood cultures for rapid identification are available. MBT Sepsityper® IVD Kit is one of them and is a CE-marked method. To comply with the IVDR directive, Sepsityper® needs to be tested and compared with the current in-house method. These two methods were compared based on the ability to provide fast and reliable results. Material and method: All the positive blood samples that were examined were prepared by the two methods then analysed with MALDI-TOF MS. The in-house-method used saponin to lyse the blood cells, Sepsityper used a lysisbuffert. How reliable the results are presented as score value, >2 means that the result is reliable to species identification, between 1.7-1.99 to genus identification and

The mesenchymal stromal cell (MSC) is of great interest for its immunomodulatory and regenerative properties. However, to research and use these MSCs it is essential to identify and characterize them as such. They need to fulfill the MSCs' minimal criteria which assess the differentiation potential, cell surface markers, and adherence. In this study, cells donated from human bone marrow were identified as MSC according to the minimal criteria. Methods used were flow cytometry, immunofluorescent staining, and ELISA. Furthermore, the population was cultured in three different media (DMEM-LG with either 10% FBS, 2% FBS, or 10% FBS supplemented with 10% conditioned media from human urinary bladder carcinoma cells (T24)) for 21 days whereupon tested for the mesenchymal characteristics, cells were counted and size measured at every passage. All cultures maintained their mesenchymal character, however, cells grown in 2% FBS became a considerably more heterogenous population regarding cell size and granularity, perhaps because of senescence. Additionally, these cells somewhat decreased in proliferation and resulted in 1 x 106 cells after 21 days, however, this was not a significant decrease when compared to the 10% FBS culture which had 2.16 x 106 cells after 21 days (p=0.061). On the contrary, the culture supplemented with T24 conditioned media resulted in a significantly higher cell count with 4.75 x 106 cells (p=0.008). Further studies could investigate which components in the conditioned media contributed to the proliferation. Moreover, the cell population in this study could not be characterized as MSC with certainty as additional cell surface markers should be tested.

Clonal haematopoiesis of indeterminate potential, or CHIP are a family of mutations present in the general population. CHIP-mutations are prevalent in the haematopoietic stem cells and in the more mature cell populations, T-lymphocytes, B-lymphocytes and myeloid cells (CD3+, CD19+ and CD33+ cells) in blood. By separating these cell populations using magnetic isolation, extracting DNA from the cell populations, and detecting the same mutation in all cell populations, one can prove the presence of CHIP-mutations in a hematopoietic stem cell. At least 50 ng good quality DNA is needed for the gene analysis to detect CHIP-mutations. The magnet separated cell population may be very small, so the DNA extraction method must be optimized to achieve enough DNA yield. The main purpose of the method development was to compare two storage methods before DNA-extractions, and then three different DNA-quantification methods after the DNA-extractions. After the best storage and quantification methods were identified, five samples of cryo-preserved viable cells were used to isolate cell populations using magnetic beads covered in specific antibodies and a magnetic field, and then quantified. Results of the study showed that the best way was to store the cells in ATL-buffer and Proteinase K. To quantify DNA, qPCR was the most accurate method, since the other methods showed incorrect results because of the low DNA concentrations. Magnet cell separation was partly successful. All except one of the DNA yields from the cell separation protocols reached the critical amount of DNA, but some yields were not pure yields of the sought-after cell population. In general, the method must be worked on more with further research.

Adipocyte signaling, normally and in type 2 diabetes, is far from fully understood. We have earlier developed detailed dynamic mathematical models for several well-studied, partially overlapping, signaling pathways in adipocytes. Still, these models only cover a fraction of the total cellular response. For a broader coverage of the response, large-scale phosphoproteomic data and systems level knowledge on protein interactions are key. However, methods to combine detailed dynamic models with large-scale data, using information about the confidence of included interactions, are lacking. We have developed a method to first establish a core model by connecting existing models of adipocyte cellular signaling for: (1) lipolysis and fatty acid release, (2) glucose uptake, and (3) the release of adiponectin. Next, we use publicly available phosphoproteome data for the insulin response in adipocytes together with prior knowledge on protein interactions, to identify phosphosites downstream of the core model. In a parallel pairwise approach with low computation time, we test whether identified phosphosites can be added to the model. We iteratively collect accepted additions into layers and continue the search for phosphosites downstream of these added layers. For the first 30 layers with the highest confidence (311 added phosphosites), the model predicts independent data well (70–90% correct), and the predictive capability gradually decreases when we add layers of decreasing confidence. In total, 57 layers (3059 phosphosites) can be added to the model with predictive ability kept. Finally, our large-scale, layered model enables dynamic simulations of systems-wide alterations in adipocytes in type 2 diabetes. CC BY 4.0© 2023, The Author(s)Correspondence and requests for materials should be addressed to William Lövfors, Gunnar Cedersund or Elin Nyman.GC acknowledges support from the Swedish Research Council (2018-05418, 2018-03319), CENIIT (15.09), the Swedish Foundation for Strategic Research (ITM17-0245), SciLifeLab National COVID-19 Research Program financed by the Knut and Alice Wallenberg Foundation (2020.0182), the H2020 project PRECISE4Q (777107), the Swedish Fund for Research without Animal Experiments (F2019-0010), ELLIIT (2020-A12), and VINNOVA (VisualSweden, 2020-04711). EN acknowledges support from the Swedish Research Council (Dnr 2019-03767), the Heart and Lung Foundation, CENIIT (20.08), Åke Wibergs Stiftelse (M19-0449, M21-0030, M22-0027), and the Swedish Fund for Research without Animal Experiments (S2021-0008). GC and WL acknowledge scientific support from the Exploring Inflammation in Health and Disease (XHiDE) Consortium, which is a strategic research profile at Örebro University funded by the Knowledge Foundation (20200017). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Background: In recent years, neuroimaging with deep learning (DL) algorithms have made remarkable advances in the diagnosis of neurodegenerative disorders. However, applying DL in different medical domains is usually challenged by lack of labeled data. To address this challenge, transfer learning (TL) has been applied to use state-of-the-art convolution neural networks pre-trained on natural images. Yet, there are differences in characteristics between medical and natural images, also image classification and targeted medical diagnosis tasks. The purpose of this study is to investigate the performance of specialized and TL in the classification of neurodegenerative disorders using 3D volumes of 18F-FDG-PET brain scans. Results: Results show that TL models are suboptimal for classification of neurodegenerative disorders, especially when the objective is to separate more than two disorders. Additionally, specialized CNN model provides better interpretations of predicted diagnosis. Conclusions: TL can indeed lead to superior performance on binary classification in timely and data efficient manner, yet for detecting more than a single disorder, TL models do not perform well. Additionally, custom 3D model performs comparably to TL models for binary classification, and interestingly perform better for diagnosis of multiple disorders. The results confirm the superiority of the custom 3D-CNN in providing better explainable model compared to TL adopted ones. Funding Agencies|Center for Applied Intelligent System Research (CAISR) at Halmstad University, Sweden; Department of Clinical Physiology, Department of Radiology; Center for Medical Imaging Visualization (CMIV) at Linkoeping University Hospital, Sweden; European DLB consortium; Analytic Imaging Diagnostics Arena (AIDA) initiative; VINNOVA; Formas [2017-02447]; Swedish Energy Agency; Swiss National Science Foundation; Velux Foundation [320030_169876, 320030_185028]; Flanders Research Foundation [1123]; [FWO 12I2121N]

Objective: The genetic predisposition to ankylosing spondylitis (AS) has been most widely studied in cohorts with European ancestry. However, within Europe, disease prevalence is higher in Sweden. Given this, we aimed to characterize known AS susceptibility variants in a homogeneous Swedish data set, assessing reproducibility and direction of effect. Method: The power to detect association within an existing Swedish targeted sequencing study (381 controls; 310 AS cases) was examined, and a set of published associations (n = 151) was intersected with available genotypes. Association to disease was calculated using logistic regression accounting for population structure, and HLA-B27 status was determined with direct polymerase chain reaction genotyping. Results: The cases were found to be 92.3% HLA-B27 positive, with the data set showing >= 80% predictive power to replicate associations, with odds ratios >= 1.6 over a range of allele frequencies (0.1-0.7). Thirty-four markers, representing 23 gene loci, were available for investigation. The replicated variants tagged MICA and IL23R loci (p < 1.47 x 10(-3)), with variable direction of effect noted for gene loci IL1R1 and MST1. Conclusion: The Swedish data set successfully replicated both major histocompatibility complex (MHC) and non-MHC loci, and revealed a different replication pattern compared to discovery data sets. This was possibly due to population demographics, including HLA-B27 frequency and measured comorbidities. Funding Agencies|Swedish Research Council, FORMASSwedish Research CouncilSwedish Research Council Formas [Dnr 2012-1531]; Wallenberg Scholar award

Significance Spatial frequency domain imaging (SFDI) and spatial frequency domain spectroscopy (SFDS) are emerging tools to non-invasively assess tissues. However, the presence of aberrations can complicate processing and interpretation. Aim This study develops a method to characterize optical aberrations when performing SFDI/S measurements. Additionally, we propose a post-processing method to compensate for these aberrations and recover arbitrary subsurface optical properties. Approach Using a custom SFDS system, we extract absorption and scattering coefficients from a reference phantom at 0 to 15 mm distances from the ideal focus. In post-processing, we characterize aberrations in terms of errors in absorption and scattering relative to the expected in-focus values. We subsequently evaluate a compensation approach in multi-distance measurements of phantoms with different optical properties and in multi-layer phantom constructs to mimic subsurface targets. Results Characterizing depth-specific aberrations revealed a strong power law such as wavelength dependence from ∼40 to ∼10 % error in both scattering and absorption. When applying the compensation method, scattering remained within 1.3% (root-mean-square) of the ideal values, independent of depth or top layer thickness, and absorption remained within 3.8%. Conclusions We have developed a protocol that allows for instrument-specific characterization and compensation for the effects of defocus and chromatic aberrations on spatial frequency domain measurements. Funding: Knut and Alice Wallenberg Foundations Center for Molecular Medicine at Linkoeping University (WCMM)

Objective. Vagus nerve stimulation (VNS) is a promising approach for the treatment of a wide variety of debilitating conditions, including autoimmune diseases and intractable epilepsy. Much remains to be learned about the molecular mechanisms involved in vagus nerve regulation of organ function. Despite an abundance of well-characterized rodent models of common chronic diseases, currently available technologies are rarely suitable for the required long-term experiments in freely moving animals, particularly experimental mice. Due to challenging anatomical limitations, many relevant experiments require miniaturized, less invasive, and wireless devices for precise stimulation of the vagus nerve and other peripheral nerves of interest. Our objective is to outline possible solutions to this problem by using nongenetic light-based stimulation. Approach. We describe how to design and benchmark new microstimulation devices that are based on transcutaneous photovoltaic stimulation. The approach is to use wired multielectrode cuffs to test different stimulation patterns, and then build photovoltaic stimulators to generate the most optimal patterns. We validate stimulation through heart rate analysis. Main results. A range of different stimulation geometries are explored with large differences in performance. Two types of photovoltaic devices are fabricated to deliver stimulation: photocapacitors and photovoltaic flags. The former is simple and more compact, but has limited efficiency. The photovoltaic flag approach is more elaborate, but highly efficient. Both can be used for wireless actuation of the vagus nerve using light impulses. Significance. These approaches can enable studies in small animals that were previously challenging, such as long-term in vivo studies for mapping functional vagus nerve innervation. This new knowledge may have potential to support clinical translation of VNS for treatment of select inflammatory and neurologic diseases. Funding Agencies|European Research Council (ERC) under the European Unions Horizon 2020 research and innovation program (EDG Grant) [949191]; city council of Brno, Czech Republic; European Research Council [834677]; Knut and Alice Wallenberg Foundation; Swedish Research Council; MedTechLabs

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.

Background/aim: We aimed to explore the roles of glycoprotein glycosylation in the pathogenesis of Kashin-Beck disease KBD , and evaluated the effectiveness of sodium hyaluronate treatment. Materials and methods: Blood and saliva were collected from KBD patients before and after the injection of sodium hyaluronate. Normal healthy subjects were included as controls. Saliva and serum lectin microarrays and saliva and serum microarray verifications were used to screen and confirm the differences in lectin levels among the three groups. Results: In saliva lectin microarray, bindings to Sophora japonica agglutinin SJA , Griffonia Bandeiraea simplicifolia lectin I GSL-I , Euonymus europaeus lectin EEL , Maackia amurensis lectin II MAL-II , Sambucus nigra lectin SNA , Hippeastrum hybrid lectin HHL , and Aleuria aurantia lectin AAL were higher in the untreated KBD patients than in the control group. Increased levels of HHL, MAL-II, and GSL-I in the untreated KBD patients discriminated them in particular from the treated ones. Jacalin was lower in the untreated KBD patients compared to the treated KBD and control groups. In serum lectin microarray, HHL and peanut agglutinin PNA were increased in the untreated KBD group in comparison to the control one. AAL, Phaseolus vulgaris agglutinin E+L PHA- E+L , and Psophocarpus tetragonolobus lectin I PTL-I were lower in the untreated KBD patients compared to the treated KBD and control groups. Hyaluronate treatment appeared to normalize SNA, AAL, and MAL-II levels in saliva, and HHL, PNA, AAL, PTL-I, and PHA-E+L levels in serum. Saliva reversed microarray verification confirmed significant differences between the groups in SNA and Jacalin, in particular for GSL-I levels, while serum reversed microarray verification indicated that HHL, PNA, and AAL levels returned to normal levels after the hyaluronate treatment. Lectin blot confirmed significant differences in HHL, AAL, and Jacalin in saliva, and HHL, PNA, PHA-E+L, and AAL in serum. Conclusion: HHL in saliva and serum may be a valuable diagnostic biomarker of KBD, and it may be used as follow-up for the hyaluronate treatment.

The introduction of novel methodologies in the past decade has advanced research on mechanisms of change in observational studies. Time-lagged panel models allow us to track session-by-session changes and focus on within-patient associations between predictors and outcomes. This shift is crucial as change in mechanisms inherently takes place at a within-patient level. These models also enable preliminary casual inferences, which can guide the development of effective personalised interventions that target mechanisms of change, used at specific treatment phases for optimal effect. Given their complexity, panel models need to be implemented with caution, as different modelling choices can significantly affect results and reduce replicability. We outline three central methodological recommendations for use of time-lagged panel analysis to study mechanisms of change: (a) taking patient-specific effects into account, separating out stable between-person differences from within-person fluctuations over time; (b) properly controlling for autoregressive effects; and (c) considering long-term time trends. We demonstrate these recommendations in an applied example examining the session-by-session alliance-outcome association in a naturalistic psychotherapy study. We present limitations of time-lagged panel analysis and future directions. Funding Agencies|NIMHUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of Mental Health (NIMH) [T32 MH019132]

Salivary pellicles display exceptional hydration and lubrication performance. At present, there are still gaps in the understanding of how this is achieved. The aim of this thesis was therefore to increase our knowledge on the mechanisms underlying these properties and deepen the understanding of how they are related to the composition and structure of pellicles, with a focus on those formed under in vitro conditions. This has applications ranging from the development of artificial saliva and lubricating coatings for biomedical applications to methodological approaches for initial testing of oral healthcare products. For this, we also focused on developing suitable methodological approaches for these studies, centering on atomic force microscopy, quartz crystal microbalance with dissipation monitoring, ellipsometry and neutron reflectometry techniques, to investigate in vitro and model salivary pellicles. First, we confirmed a two-layer structure for in vitro salivary pellicles and showed that the outer layer is mainly composed by the oral mucin MUC5B, but that it also contains other salivary components that enhance swelling and hydration. In the presence of bulk saliva, the outer layer also contains a reversibly and loosely bound fraction. This fraction increases the adhesiveness of the pellicle but unexpectedly has no significant effect on its lubrication performance. We also investigated the effect of mechanical confinement on model salivary pellicles by means of Neutron Reflectometry, revealing that at a pressure of 1 bar they are already completely compressed and dehydrated. Finally, with the aim to advance towards better oral healthcare products, we investigated the effect of nonionic and amphoteric surfactants on salivary pellicles, showing that they have a gentler effect on pellicle structure than the commonly employed anionic surfactants.

A mobile and web-based prototype was developed to explore utility of heart sound data in the context of patient self-monitoring. There are not many applications available despite measurement equipment that can be purchased. This research aimed at developing an application that could help patients understand and use phonocardiography. The resulting prototype Intellicor enables easy-to-use web and mobile solutions such as registering heart sound, review of previous heart signal recordings, summaries of terms related to patient condition, and medication taken. These functions can be utilized by both patients and physicians to create understanding of heart signals and build communication as a part of treatment. Three development iterations included several expert evaluators who gave positive feedback on the concept of the application. It was appreciated that patients could monitor heart signals and better understand the results. The medical experts would welcome such a system into their work domain if developed correctly and in accordance with the formal expectations, both legal and clinical. The prototype has shown the advantage of gathering data otherwise impossible to obtain. The Intellicor prototype presents the foundation that ought to be further developed in close cooperation of clinical and biomedical experts. The self-monitoring of this kind could benefit patients and the healthcare sector as demonstrated by the Intellicor prototype.

Ultraljudsundersökning av arteria carotis interna (ICA) och arteria carotis externa (ECA) utförs på fysiologiska kliniker för att bland annat bedöma blodflödeshastigheten i kärlen. Det kan i sin tur användas vid diagnostik av plack och stenoser. Behandling av stenos är viktigt eftersom det kan leda till stroke och transitorisk ischemisk attack (TIA). Syftet med den här studien är att jämföra den maximala blodflödeshastigheten i ECA och ICA, samt om den skiljer sig mellan höger och vänster sida. I studien inkluderades 36 deltagare i åldern 20–27 år. Resultatet analyserades med ett parat t-test samt med en korrelationsanalys. En statistisk signifikant skillnad identifierades vid jämförelse av ICA och ECA på samma sida. Ingen signifikant skillnad observerades vid sidojämförelse av vänster och höger ECA respektive ICA. En korrelation påvisades mellan vänster och höger ECA samt mellan vänster och höger ICA. Ytterligare en signifikant korrelation identifierades mellan ECA bilateralt och ICA dx. Slutsatsen med studien är att det finns en statistisk skillnad mellan flödeshastigheten i ICA och ECA och därav går det inte att använda samma referensvärden till båda kärlen. Ultrasound examination of the internal carotid artery (ICA) and the external carotid artery (ECA) is performed in physiological clinics to assess the blood flow velocity in the vessels. Furthermore, it can be used in the diagnosis of plaque and stenoses. Treatment of stenosis is important as it can lead to stroke and transient ischemic attack (TIA). The purpose of this study is to compare the maximum blood flow velocity in ECA and ICA and allocate a possible difference between the right and left side. The study included 36 participants between the ages of 20–27. The result was analyzed using a paired t-test and a correlation analysis. A statistical significance was identified when comparing ICA and ECA on the same side. No significant difference was found when comparing left and right ECA and ICA. One correlation was detected between left and right ECA and between left and right ICA. Another correlation was identified between ECA bilaterally and ICA dexter. The conclusion of this study is that there is a significant difference between the flow velocity in ICA and ECA and therefore it is not possible to use the same reference values ​​for both vessels.

Chromogranin A (CgA) and its cleavage product Catestatin (CST) are two very interesting proteins, especially due to their possible involvement in the development of type 1 diabetes. There are currently only a few primary antibodies against CgA and CST, and the ones that do exist are often only recommended for use on one method. Recently though, two new multifunctional antibodies were released, which are said to function on multiple methods. The aim of this project was to validate these two antibodies, one being the Chromogranin A Polyclonal Antibody (PA5-35071) from Thermo Fischer Scientific and the other being the Catestatin Polyclonal Antibody (289-MM-0288) from MediMabs. The methods of validation for these antibodies were western blot, flow cytometry, and immunofluorescent staining of frozen tissue. All study material were from mice and the tissues were chosen due to their established expression of CgA and CST. The results show that the antibodies are highly appropriate for use on flow cytometry and immunofluorescent staining of frozen tissue. The most appropriate concentration for the CgA-antibody was deemed as 1:25 for both flow cytometry and immunofluorescent staining. The most appropriate concentrations for the CST-antibody were deemed as 1:800 for flow cytometry and 1:1000 for immunofluorescent staining. However, the antibodies could not be validated for use on western blot. Further investigations are needed to determine if the antibodies are functional in western blot analysis or not. The validation performed here is an important first step in studying the involvement of these proteins in type 1 diabetes onset.

Sleep is very important for health and academic performance. The amount of sleep is variable and depends among other things, on the age of the individual. Today there are many varieties of motion sensors that can be used in healthcare and clinical research for sleep registration, such as Actigraph GT9X Link and Actiwatch 2. The purpose of this study was to compare these two sensors, Actigraph and Actiwatch, for sleep registration by analyzing different sleep variables. The more precise sensor of the two will be used in the Energymetabolic laboratory in the case of survey or research. The method involved participants wearing both sensors for two nights and one day and trying to live as normal as possible. A sleep diary was used to complete the registrations, with related questions about sleep. There were 17 healthy participants between the ages 21 and 31. The results showed that the registrations from both sensors have varying spread and deviations. Overall the results were similar between the sensors and the sleep diary, and it was shown that there is no significant difference between the sensors. However, Actigraph deviated on most occasions from the diary. The Energymetabolic laboratory have access to many Actigraph GT9X Link sensors. This means that an evaluation has to be made to decide whether the cost of replacing the Actigraph GT9X Link sensors with Actiwatch 2 is worth it for the small improvement in precision.

Objectives Proposal of a risk analysis model to diminish negative impact on patient care by preanalytical errors in blood gas analysis (BGA). Methods Here we designed a Failure Mode and Effects Analysis (FMEA) risk assessment template for BGA, based on literature references and expertise of an international team of laboratory and clinical health care professionals. Results The FMEA identifies pre-analytical process steps, errors that may occur whilst performing BGA (potential failure mode), possible consequences (potential failure effect) and preventive/corrective actions (current controls). Probability of failure occurrence (OCC), severity of failure (SEV) and probability of failure detection (DET) are scored per potential failure mode. OCC and DET depend on test setting and patient population e.g., they differ in primary community health centres as compared to secondary community hospitals and third line university or specialized hospitals. OCC and DET also differ between stand-alone and networked instruments, manual and automated patient identification, and whether results are automatically transmitted to the patient’s electronic health record. The risk priority number (RPN = SEV × OCC × DET) can be applied to determine the sequence in which risks are addressed. RPN can be recalculated after implementing changes to decrease OCC and/or increase DET. Key performance indicators are also proposed to evaluate changes. Conclusions This FMEA model will help health care professionals manage and minimize the risk of preanalytical errors in BGA.