Den tilltagande höga nivån av urbanisering och vägbyggen är en bidragande faktor till spridning av invasiva arter. Transporter och frekventa vegetationsstörningar resulterar i homogena, öppna och soliga landskap med minskad konkurrens där snabbt växande ruderala arter med hög ihållande fröproduktion etablerar sig, varav många är invasiva. Syftet med projektet är att undersöka spridningsfaktorer av invasiva växtarter inom c-verksamheter som återvunnit jordmassor mellan 2020–2021 inom Göteborg stad, Kungälv- och Härryda kommun. Dyker invasiva arter upp i samband med flytt av jordmassor? Utgör verksamhetsstrukturer med sina öppna ytor en spridningskorridor för redan befintliga arter i området? Vilken av verksamheterna bedöms utgöra störst risk för spridning? Invasiva arter (SE) inventerades inom sju kartlagda C-verksamheter (bullervallar, markhöjning på jordbruksmark, banarbete på motocrossbana och golfbana) med en buffert på 250 meter. Artförekomst noterades med värdet 1 för närvaro och värdet 0 för frånvaro. Små plantor av invasiva arter kunde även indikera på etablering efter masshantering. I QGIS 3.24 beräknades avståndet mellan förklaringsvariablerna vägar, stigar, järnvägar, vattenförekomst (NMD) och övriga marktäckedata (NMD). Alla bearbetade data exporterades till Rstudio för att utveckla en logistisk regressionsmodell (GLM) i syfte att förutspå invasiv artförekomst utifrån förklaringsvariablerna. Detta var nödvändigt då en logistisk GLMM inte visade någon skillnad mellan verksamheterna. Resultatet från regressionsanalysen (GLM) visade att det fanns ett samband mellan artförekomst och variablerna Vägar, Stigar och hantering av jordmassor. Detta beror delvis på att vägar och stigar öppnar upp för mänskliga aktiviteter som gynnar fröspridning. Verksamheter som bullervallar och motocrossbanor är starkt anknutna till vägstrukturer och mänskliga aktiviteter. Framtida studier bör inkludera förklaringsvariabler som befolkningstäthet inom verksamhetsområdet, nettoprimärproduktion, slänter, jordtyp och dess fröinnehåll, då även dessa variabler har visat statistisk signifikant förutsägelse om artförekomst i tidigare studier. För ökad statistisk styrka kan ett högre antal relevanta verksamheter av flera sorter undersökas, innefattande deponier, sand och grustag. / The project aimed to investigate dispersal factors of invasive plant species within C-operations, which recycled soil masses between 2020–2021 in the municipalities of Gothenburg, Kungälv and Härryda. Do invasive species appear in connection with the movement of soil masses? Do operationstructures constitute a dispersal corridor for already existing species in the area? Which of the activities is considered to pose the greatest risk of spread? A field inventory of invasive plant species (SE) were conducted within seven mapped C-operations (noise barriers, land elevation on agricultural land, course work (motocross course and golf course) with a buffer of 250 meters. Species occurrence was noted along a transect with the value 1 for presence and the value 0 for absence. Newly sprouted plants could also indicate establishment after mass handling of soil. In QGIS the proximity between the explanatory variables roads, paths, railways, water body (NMD) and other ground cover data (NMD) was calculated in order to develop a logistic generalized linear model (GLM) to predict invasive species occurrence based on the explanatory variables. The results from the regression analysis (GLM) showed that there was a connection between species occurrence and roads, paths, noice barriers and course work that handled soil masses. This is partly because roads and paths open up for human activities that promote seed dispersal. Activities such as noise barriers and motocross tracks are strongly linked to road structures and human activities. A GLMM- analysis did not show any difference between the operations. Therefore, in future studies a higher number of relevant operations of several types can be implemented, including landfills, sand and gravel pits. Also, explanatory variables such as population density in the area, net primary production, slopes, soil type and its seed content, as these variables have shown statistically significant predictions of species occurrence.

Sepsis is an irregular systemic response to an infection, in which a pathogen or some of its component(s)reaches the bloodstream of the host or sterile tissue, triggering a disproportionate immune reaction. The first three hours are critical in the diagnosis of sepsis, in order to ensure an effective treatment with less impact on the patient. Culture-dependent diagnosis is the present standard procedure which can take up to several days. Metagenomics Next Generation Sequencing (mNGS) is a culture independent diagnostics method which could be used to identify the presence of pathogens from DNA extracted from human whole blood enabling a more effective treatment procedure of infected patients. The aim of this research was to utilize the sequencing data obtained with the MinION Nanopore sequencing device, in order to systematize its use as a tool for the early detection of sepsis; furthermore, determine if this technology is effective to use on DNA extracted from whole blood. The main research question of this thesis focused on whether the MinION Nanopore sequencing is a reliable tool for the early detection of sepsis. Whole blood samples from healthy donors was spiked with bacteria and DNA was extracted and sequenced with MinION device. The sequencing results were interpreted with the MinKNOW v2.0 software, through the application What’s In My Pot (WIMP). Also, the web tool PATRIC 3.6.12. and KRAKEN2 algorithm. The reads from the taxonomic family where the bacteria belong to was analyzed, presuming the bacterial DNA was present in the DNA extracted but the genus was not detected. According to the KRAKEN2 and WIMP analysis, the bacteria used to spike the whole blood samples was detected up to the taxonomic family level. Thus, confirming the presence of the spiked bacteria in the purified DNA samples.

Neuroblastomas are the most common extra-cranial abnormal growths in immature nerve tissue within the sympathetic nervous system, in children ages 0-5. It is a heterogeneous malignancy, often emerging in and around adrenal glands, caused by rapid growth and division of nerve cells. From a clinical perspective it is difficult to treat as it has spontaneous regression, rapid progression and therapy resistance. MicroRNAs (miRNA) are small, single stranded RNA, 18-25 nucleotides long and act as human gene expression regulators which are deregulated in Neuroblastoma. The expression of miRNAs can be analysed to determine the progressive stage at which the neuroblastoma is, and the best course of treatment can be decided. The aim of this study was to analyse the expression of miRNA hsa-miR-34c-5p, in three different cell lines each having different genetic characteristics, NB69 without MYCN amplification and 11 q deletion, SN-BE with MYCN amplification and Kelly with 11q deletion. As hsa-miR-34c-5p has been found to have tumour suppressive capabilities in Neuroblastoma cell lines. The cell lines were cultured, the total RNA was isolated, Reverse Transcription was done to convert small RNA to cDNA and qPCR was run and Cq values were obtained. The miRNA expression was determined using qPCR and Livak´s method, however, no statistical difference was found in the expression of hsa-miR-34c-5p between the Neuroblastoma cell lines. These results are only a steppingstone in future research of miRNA and Neuroblastoma.

Inflammation is a part of the innate immune system. It protects the body against foreign invaders such as bacteria and viruses. Inflammation helps to restore the body by removing harmful stimuli and starting the healing process. Inflammation is produced in response to damage-associated molecular patterns (DAMPS) or pathogen-associated molecular patterns (PAMPS). Glycogen synthase kinase 3 (GSK-3) is a key regulator of a variety of pathways, making it a promising therapeutic target. Therefore, this experiment aims to see how inhibiting GSK-3 affects the generation of IL-8 in THP-1 ASC GFP macrophage cells. For this study qPCR was used to measure IL-8 expression, while ELISA was used for protein secretion. An ANOVA test was utilized for the statistical analysis. Obtained results from this study showed that there is a significant difference between stimulated cells with LPS and nigericin against unstimulated samples both in protein and mRNA levels. When it comes to the stimulated cells against inhibited cells, the ANOVA test showed there is no significant difference between the samples both in protein and mRNA levels. This might suggest that GSK-3 does not influence the development of inflammasomes in THP-1 macrophage cells. Another possible reason is that other pathways such as the MAPK and JAK-STAT may mask potential inhibitory effects on the NLRP3 inflammasome pathway by producing even more IL-8, which interfered with qPCR and ELISA results. In conclusion, additional research is needed to confirm the involvement of GSK-3 in NLRP3 inflammation.

The testis is a complex organ responsible for sperm production and hormone synthesis. Spermatogenesis, the process of sperm formation, occurs in the seminiferous tubules and involves the coordinated development of germ cells with the support of Sertoli cells. These provide structural and nutritional support for germ cells, regulate spermatogenesis, and maintain the blood-testis barrier. Despite extensive research on Sertoli cells, their protein expression patterns and spatial localization in relation to other testicular cell types remain poorly characterized. This project aimed to investigate the expression patterns and spatial localization of proteins in Sertoli cells. Single-cell RNA sequencing data and immunohistochemistry stainings from the Human Protein Atlas were used to choose candidate proteins to be visualized in a multiplex Immunofluorescence panel. After staining, the slides were scanned and analyzed manually as well as through automated image analysis via QuPath. 101 proteins were successfully stained and localized within testis tissue and subcellular locations in Sertoli cells were annotated. Additionally, a list of uncharacterized proteins in Sertoli cells were identified. The employed workflow demonstrated its potential for identifying Sertoli cell-specific proteins and can be applied to future research. Additional validation is required to verify the presence and identify function of these proteins in Sertoli cells. For this purpose, techniques such as mass spectrometry-based proteomics and mouse knock-out models could be used.

Hepatitis C viral infection is a globally widespread blood-borne disease affecting the liver, causing cirrhosis and hepatocellular carcinoma. This type of liver cancer is mainly caused by chronic Hepatitis C and Hepatitis B. Specific detection with the following correct treatment is crucial to reduce the overall burden of the disease. This work focused on investigating whether the padlock probes and rolling circle amplification can detect Hepatitis C, determining the limit of detection, and if any blood components would inhibit the reactions. All oligonucleotides were tested for ligation functionality in 10% TBE-Ureal gel electrophoresis and used with rolling circle amplification and phi29 polymerase to determine if eye read-out was possible. The lowest concentration of detection was found to be 10 pM. To avoid inhibition in blood plasma, samples were pre-treated at 95 ˚C for five minutes. Eye read-out was possible after amplification, with 30% plasma at the highest and 5% plasma at the lowest in samples. In conclusion, this novel assay using padlock probes, a detection oligonucleotide, and rolling circle amplification holds promise in developing a simplified new detection technique for the diagnostics of Hepatitis C.

Trimethylaminuria is an autosomal recessive disorder characterized by a decreased oxidation capacity of trimethylamine to trimethylamine-N-oxide in the liver. The condition is diagnosed by estimating concentrations of trimethylamine and trimethylamine-N-oxide in human urine and then evaluating their respective creatinine ratios and the oxidation efficiency percentage. Values previously retrieved with liquid chromatography-tandem mass spectrometry with electrospray ionization but not the novel ionization interface UniSpray. Thus, this project aims to initiate the development of a liquid chromatography-tandem mass spectrometry diagnostic method for trimethylaminuria with UniSpray ionization. The analytes were extracted from urine with a liquid-liquid extraction method and separated with hydrophilic interaction liquid chromatography using an isocratic profile with 5 mM of ammonium formate in water and methanol and multiple reaction monitoring. Overall, the mean coefficient of variation and recovery percentage from trimethylamine spiked urine samples were lower than expected, whereas the intra-precision for trimethylamine-N-oxide was acceptable. Three urine samples had estimated oxidation percentages, but only one had derived comparable creatinine ratios to an external laboratory. Due to the inadequacy of comparative data and the precision and recovery percentage deviations, the results presented in this report need cautious interpretation. Future development of the method could include manual tuning, reconsidering the calibration curve and reference values, and comparisons to the extraction method. Although there are apparent discrepancies in the precision and reliability of the derived trimethylamine and trimethylamineN-oxide concentrations, the initial steps in the pursuit of a liquid chromatography-tandem mass spectrometry diagnostic method for trimethylaminuria provide a practical foundation to continue the development.

Inflammation is the body's natural defense reaction and is known since ancient times. The inflammation is divided into two main phases, acute and chronic inflammation dependent on the process and cellular mechanisms of the inflammation. Inflammation has become to be an important field in research by biomedical research where it is included in many cellular processes thus being phagocytosis, chemotaxis, mitosis, and cell differentiation. Inflammasomes are pro-inflammatory intracellular multimeric protein complexes that introduce the activation of pro-inflammatory cytokines, such as interleukin-1β and interleukin-18, upon trigger by PAMPs and DAMPs signals. The most studied inflammasome is the NLRP3 inflammasome that is activated by various trigger signals, like DAMPs, ATP, uric acid crystals and amyloid-β fibrils. GSK-3β is a kinase that controls various cellular processes, such as inflammation by regulating the activity of abundant transcription factors that are valuable for cytokine production. The aim of this thesis project was to investigate if GSK-3 Inhibitor IV, SB-216763, in a concentration-dependent manner had an effect on production of IL-1β in LPS- and Nigericin-stimulated THP-1 ASC-GFP-macrophages. In addition to the gene expression analysis of IL-1β, the amount of secreted IL-1β, and the possible correlation between treated THP-1 cells with and without GSK-3 inhibitor evaluated. The gene expression analysis was performed by using qPCR and the amount of secreted IL-1β was done using sandwich enzyme-linked immunosorbent assay. The results from this study showed no significant difference in gene expression and amount secreted of IL-1β in THP-1 cells when treated with the GSK-3 Inhibitor IV, SB-216763.

Corona virus disease caused by severe acute respiratory virus 2 causes blockage of the blood vessel which leads to thrombosis. Thrombotic events in covid-19 patients results to hospitalizations and death. And incidence of thrombosis in covid-19 patients have been increasing in most regions of the world. Due to the huge inequalities between developed and developing countries, incidence rates remain highest in more developed regions, but mortality is relatively much higher in less developed countries due to a lack of early detection and access to treatment facilities. The aim of this study was to investigate the incidence of thrombosis in covid-19 by performing a systematic review on research articles talking about thrombotic events in covid-19 patients, carrying out a meta-analysis on the generated data to make an inference on the statistical result in order to create an information concerning complications covid-19 patients suffer from. Literatures with cases of covid-19 that reported D-dimer elevation and that followed the WHO standard for covid-19 diagnosis were included. Literatures excluded were studies with pregnant women, cancer patients and patients undergoing chemotherapy and with no approved ethical considerations. Information sources included only original literatures with initial search yielding 55 results from 4 databases. After reviewing titles and abstracts of all 55 literatures, 35 studies were further screened and 10 were included in the analysis representing 3359 patients. A forest plot using the R programming language was done, and an overall pooled estimate using the random effect model was 20 % (95 % confidence interval 12.0 % - 29.0 %) with heterogeneity of 96 %, and p <0.01. The incidence of thrombosis among moderate cases of c ovid-19 patients was 12 % with (95 % confidence interval 8.0 % - 18.0 %) with heterogeneity 93 %, and the incidence of thrombosis among severe Covid-19 patients was 22 % (95% confidence 10.0 %-37.0 %) with 97 % heterogeneity, and p <0.01.

Sclerotinia sclerotiorum is a plant pathogenic fungus that causes Sclerotinia stem rot in oilseed rape. In Sweden, the disease causes severe crop loss that varies by year. Previous studies have shown a relationship between the proportion of infected petals and disease incidence in infected fields in places with high humidity levels before and during flowering. In this study, the aim was to develop a technique to detect S. sclerotiorum and other fungi pathogens in the petals of oilseed rape from naturally infected fields by using nanopore sequencing from Oxford Nanopore Technologies. DNA was extracted from the petals of oilseed rape and subsequently amplified by performing PCR after optimizing the optimal annealing temperature. Using the forward primer ITS1catta and the reverse primer ITS4ngsUni, these primers targeted the ITS region, which is used as a marker for the identification of fungi. The resulting Amplicon concentrations varied. Five amplicon PCR samples were selected for MinION sequencing. These samples were selected since they had the best purity levels. Finally, bioinformatic analysis was done with Kraken2 and the Pavian tool and compared with UNITE databases. The result showed hundreds of thousands of reads were recovered from the Ascomycota and Basidiomycota fungi divisions; S. sclerotiorum was observed in one field sample; other Sclerotiniaceae species like Dumontinia tuberosa, Botrytis cinerea, and Sclerotinia bulborum were detected in two fields; and many other fungal pathogen species affecting rapeseed crops in Sweden were successfully detected. MinION was successful in identifying S. sclerotiorum and other plant pathogens.