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Bakgrund: Vitamin D är ett steroidvitamin och tillförs oss från vissa livsmedel eller syntetiseras när vår hud utsätts för solen. Inuti cellerna fungerar det som en transkriptionsfaktor för en grupp gener. 1,25-dihydroxivitamin D3 (1,25D3) är den aktiva vitamin D-metaboliten, men den cirkulerar mest i kroppen som 25-dihydroxivitamin D3 (25D3). ER-stress uppstår när proteinsyntesen inte fungerar för att försöka rädda cellen. Tidigare studier har föreslagit en skyddande effekt av vitamin D på ER-stress.  Syfte: Syftet med denna studie var att undersöka effekterna av metaboliter av vitamin D på ER-stress i PC3 och NSC34-celler. Metoder: PC3-celler och NSC34-celler behandlades med 1,25D3 eller 25 D3 i kombination med ER-stressorn tunicamycin. Stressnivåerna mättes med realtids-PCR med BiP som biomarkör för ER-stress och CYP24A1 som biomarkör för D-vitamininduktion. Resultat: Resultaten från qPCR-analysen var varierande och motsägelsefulla i de behandlade PC3-cellerna. Den genomsnittliga dubbla skillnaden i uttrycket av BiP för tunikamycin-behandlade celler var endast något större än för celler behandlade med en kombination av tunicamycin och 1,25D3 eller 25D3. När det gäller NSC34-celler verkar kombinationen av tunicamycin och en vitamin D-metabolit öka ER-stressen. När celler behandlades med tunicamycin minskade metabolismen av D-vitaminmetaboliterna. Slutsats: Denna studie ger information om inte bara effekten av den tidigare studerade 1,25D3 på ER-stress, utan även effekten av 25D3. Studien antyder också en tidigare okänd biologisk funktion av 25D3, vilket utmanar tidigare antaganden om en inaktiv metabolit.

Background: The last decades, advances in drug development have resulted in many biological drugs being launched on the market. There is limited knowledge of how they are being used. The aim of this study was compare volumes and expenditure of biologics in between 2017 and 2021 to get an overview of the most used biologicals in Stockholm, Sweden.   Methods: Descriptive cross-sectional analyses were made om data for region Stockholm on utilization and expenditure of biologics 2017 and 2021. Data were collected using databases on dispensed drugs and hospital sales from Socialstyrelsen and Region Stockholm, respectively. Measurement units used in the study were Defined Daily Doses, amount of sold packages and total expenditure in Swedish crowns.  Results: Volumes of biologics have increased over time.  Also cost of most biologic drugs increased except for the cancer drug trastuzumab. TNF-a inhibitors were the biologics accoutning for the highest cost. Some of the drugs were sold either by prescription or hospital sales only.   Conclusion: Volumes and expenditure of biologics increase in use and belong to the top-drugs by expenditure. Further research and analyses are needed to assess the appropriateness.

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Peas are small spherical seedpod of the pod fruit Pisum sativum. It is of great nutritional value containing vitamins, carbohydrates, proteins, and several minerals. A variety of diseases affects peas through a number of pathogens including insects, viruses, bacteria and fungi. Fungi and fungi like organism (oomycetes) are eukaryotic organisms known to be detrimental to plants. They are inconspicuous due to their small sizes and cryptic lifestyle in soil and are often difficult to detect. DNA barcoding uses sections of a gene to identify unknown species or parts of an organism. The official DNA barcoding marker for specie level identification of fungi, the nuclear ribosomal internal transcribed spacer ITS, has shown to contain less intraspecific variations than interspecific variation making it suitable for most fungal studies. In this study, DNA extracted from six naturally infected pea fields’ samples were amplified using two different primer pairs. ITS1Catta & ITS4ngsUni targeted fungi phylum; Ascomycetes, Zygomycetes and Basidiomycota while ITS1OO & ITS4ngs targeted phylum Oomycota. MinION sequencing was used to read the amplicons. Further taxonomic identification was done using the Kraken2 bioinformatics tool. Several reference databases were explored like UNITE, which contains fungal ITS, NCBI which covers plants, bacteria and fungi. The primer pair ITS1OO & ITS4ngs recovered 4,195 oomycetes reads while the primer pair ITS1Catta & ITS4ngsUni recovered 1983 ascomycetes and 370 basidiomycetes. MinION provides a fast and efficient alternative to current methods of identifying pea root rot causing fungi and oomycetes.

Sepsis is a life-threatening condition in which a dysregulated immune response causes damage to the patient’s organs. The condition moves fast leaving little time for diagnosis and starting treatment. Early diagnosis would help save lives and reduce the amount of antibiotics used. The aim of this study was to compare manual and robotic extractions for miRNA extraction and to quantify miR-seps4, a possible biomarker for sepsis, from human plasma. During the first part of the study manual extractions done by hand and robotic extractions done using QIAcube (Qiagen) of miRNA using the miRNeasy Serum/Plasma Advanced Kit (Qiagen) were compared using the quantity and purity of RNA they produced. Then two-tailed RT-qPCR was optimised for human plasma spiked with synthetic miR-seps4. The final objective was to use these methods to test whether two-tailed RT-qPCR could be used to identify and quantify miR-seps4 from non-spiked human plasma using the absolute quantification method. The QIAcube (Qiagen) extractions gave a higher concentration of RNA while manual RNA extractions had a higher purity A successfully optimised standard curve was made using synthetic miR-seps4 though the melt curve showed contamination and possible non-specific products. Non-spiked RNA extractions showed successful amplification of miR-seps4 that could be quantified through the absolute quantification method. This makes miR-seps4 a good target for further research and a possible addition to a multi-marker panel for early diagnosis of sepsis.

Sepsis is a syndrome developed in response to infection when a body’s immune system cannot adequately respond to infection. To this day, sepsis holds a peak position in mortality rates worldwide. Currently, SOFA score and blood culturing as means to diagnose sepsis are standard. However, blood culturing is lengthy and sepsis can develop at a faster rate. Faster diagnostic of sepsis in form of biomarker identification directly from the blood is one possible solution. One of the possible biomarker candidate is miRNA, a small non-coding RNA that participates in the regulation of many biological mechanisms of the human body. The aim of this work was to improve extraction of small RNA including miRNA from patient blood plasma via manual extraction using miRNeasy Advanced Kit (Qiagen) and robotic extraction using the same kit and QIAcube machine while comparing two methodologies. All samples containing extracted small RNA including miRNA were analysed with a Qubit 4.0 (Thermo Fisher Scientific) and DS-11+spectrophotometer (DeNovix). Furthermore, two-tailed RT-qPCR was investigated for potential to quantify synthetic and naturally occurring miRSeps-5 extracted from blood plasma. PCR System Machine was used to obtain Cq-values and melting curves. In order to use absolute quantification to quantify microRNA standard curve was optimized achieving a 116% efficiency and correlation coefficient R2=0.97. This work shows encouraging results of using two-tailed RT-qPCR to detect miRSeps-5 extracted from human plasma for future biomarker research.

Denna studie granskar tre länder; Tyskland, Spanien och Danmark, som använt feed-in-tariffs (FIT) under en längre tid för att främja elproduktion från solceller. Granskningen omfattar hur användningen sett ut och förändrats, samt om det resulterat i en ökning av solceller. I granskningen ingår också en genomgång av det nuvarande stödsystemet för solceller i Sverige, huruvida det främjat industrin för solceller och om FIT:s skulle kunna användas i Sverige. Granskningens resultat visar att det är svårt att härleda användningen av en viss sorts stödsystem till en ökning av solceller, då det ofta är många olika incitament som används samtidigt. Dessutom kan utformningen av FIT:s skilja sig så mycket att bara användningen av FIT inte garanterar att mängden solceller kommer öka. För att FIT:s ska kunna användas för att främja solcellskapaciteten måste tariffen vara relativt hög, jämfört med exempelvis vindkraft, samt reformeras allteftersom för att hantera de problem som dyker upp på vägen. I Sverige, som sedan 2003 använt elcertifikat och ett investeringsstöd (sedan 2005), har ingen stor ökning av solcellskapaciteten skett jämfört med de andra länderna. Teoretiskt sett kan FIT:s användas i Sverige, men det kräver politisk beslutsamhet, och en justering av elcertifikatssystemet skulle vara mindre komplicerad. / This study analyses three cases where feed-in-tariffs (FIT) have been used, and whether it resulted in an increase of solar cells (PV). The cases were Germany, Spain and Denmark and the examination of these cases included how the use of FIT: s have appeared and changed. Then the current Swedish support system for PV was studied, to see whether the promotion has been successful, and lastly if an FIT-system could be brought into place in Sweden. The results showed that, firstly, it will be hard to trace the use of just one incentive to an increase of PV-installations since most often many incentives are used at once. Secondly, the design of a FIT-system can differ so much that the sole use of FIT: s doesn’t guarantee that PV will grow. For there to be a rise in PV, the tariff must be set relatively high, compared to wind power, and continuously reformed as new problems arise along the way. In Sweden, which since 2003 have used green certificates and an investment aid (since 2005), no significant increase of PV has occurred compared to the other countries. The discussion showed that even though a FIT-system theoretically can be used in Sweden it demands political determination, and that an adjustment or reformation of the certificate system would be less complicated.

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