Neuroendocrine tumors (NET) are an unusual form of cancer that can occur in many different organs, commonly in the small intestinal. NET can be detected with the help of immunohistochemistry (IHC). IHC uses antibodies that bind to an antigen in the tissue. NET can be studied through this bond, which forms a colored product during an enzyme reaction, using a chromogenic or fluorescence-based method. Usually, NETs are analyzed with synaptophysin, chromogranin A, and CD56 markers. Novel studies have been conducted on insulin-associated protein 1(INSM1), a new emerging NET marker suggested to have higher specificity and sensitivity than the markers mentioned above.   The work aimed to investigate the optimal protocol for INSM1, test an appropriate control, and evaluate if the INSM1 antibody works according to the description in the scientific literature and is suitable to be introduced into the routine practice and used as a diagnostic marker at the department for pathology and cytology in Växjö.  The evaluation of INSM1 was performed on sectioned tissue samples with NET. NET staining was evaluated by dilution of the INSM1 antibody, different pretreatments and visualizing kits were used. The staining’s of INSM1 were assessed microscopically together with a pathologist.   The analysis showed that the optimal result was gained by using 1 mg/ml of the INSM1 antibody. The staining intensity was preferable using the OptiView compared to UltraView. The INSM1-antibodies showed better binding to the antigen when using the “standard” pretreatment and the color intensity was more intense compared to the “mild” pretreatment. The evaluation showed that OptiView and standard pretreatment, gave the optimal result for NET diagnostics using the INSM1 antibody. Based on the work herein, the antibody INSM1 could be introduced into the routine practice at the department of pathology and cytology in Växjö.

Stem cell niches serve as vital reservoirs, maintaining stem cells in a dormant state through specific niche components. Various biomarkers associated with stem cell niches have been identified in different tissues, including cardiac tissue, particularly at the atrioventricular junction. This study collected mouse, donor, and explanted cardiac tissue from the right side of the heart, focusing on the atrioventricular junction. Additionally, left ventricle samples from donor hearts were harvested. All samples were examined via immunohistochemistry to detect niche-related biomarkers. The study aimed to validate the atrioventricular junction as a potential niche area and to explore Cathepsin K asa possible cardiac stem cell marker, noting its slight nuclear expression. Clear expression of the stemcell marker SSEA4, the hypoxia marker Hif-1α, and the extracellular matrix marker ACAN, along with weak expression of the extracellular matrix marker CILP2, was observed, alongside Cathepsin K. Notably, explanted cardiac samples exhibited lower marker expression, except for Hif-1α, suggesting reduced oxygen levels in failing hearts. Mouse samples displayed the highest expression for each marker, mirroring the distribution seen in human hearts. Although the study does not conclusively validate the existence of a cardiac stem cell niche or the role of Cathepsin K, its findings significantly advance niche research. Further investigation is warranted to elucidate the role of the atrioventricular junction and associated biomarkers in cardiac stem cell biology.

Liver Cancer, especially hepatocellular carcinoma is a major global health challenge with increasing occurrence and mortality rates. This study aimed to explore and understand the molecular signature of the disease using bioinformatic analysis with The Cancer Genome Atlas Liver Hepatocellular Carcinoma dataset to identify differentially expressed genes and somatic mutations associated with patient survival. Transcriptome analysis revealed a significant difference in gene expression between tumor and normal tissues. Survival analysis linked specific genes to patient outcomes which suggests potential prognostic markers further proved by clinical enrichment analysis using Metascape, an online platform. Single nucleotide variant analysis uncovered frequently mutated genes, including TP53, CTNNB1, and TTN, and characterized the mutation landscape, revealing prevalent C>T transitions. The integration of transcriptome and mutation data complements potential therapeutic targets and driver mutations which are important for the liver cancer development and progression. Overall this study provides valuable insights and is a framework into the molecular mechanisms of HCC and identifies potential DEGs, biomarkers (FCN2, FCN3, and COLEC10), and clinically actionable targets for improved diagnosis and therapeutic strategies.

Avian influenza, known as bird flu, is a highly contagious acute respiratory infection that infects many avian species, humans and other animals. Birds infected with Influenza A virus can shed large amounts of virus in feces, which can be detected in the lakes. Similarly, people infected with enteric viruses also shed high amounts of virus in their feces, which can end up in lake water. In this study, the presence of influenza A and seven enteric viruses (norovirus GII, astrovirus, enterovirus, adenovirus, sapovirus, hepatitis E virus, and pepper mild mottle virus) were investigated in recreational lakes to explore whether their identification could be used as an early warning of outbreaks. Lake water samples were collected from Axlemossen, Delsjön, Slottsskogen, and Torpadammen. The samples were taken monthly from November 2023 until April 2024 and analyzed for detection of viruses by concentration through ultrafiltration and quantitative polymerase chain reaction. Influenza A virus was selectively detected in the urban lakes of Slottsskogen and Torpadammen, where favorable pH, temperature, and high bird populations likely supported the virus's stability in the lakes. In contrast, enteric viruses showed a broader distribution across all lakes, likely driven by human fecal contamination from recreational activities or possibly from untreated wastewater discharges from unknown sources. The method used in this study aids in studying the presence and behavior of circulating strains in birds. Additionally, it can help understand the potential public health risks of waterborne viruses, especially those transmitted through avian or human fecal matter in the local environment.

The zone of Ranvier has been identified as a stem cell niche located directly next to the epiphyseal growth plate in the knee structure, which has drawn attention as a potential area of study in stem cell therapy treatments for degenerative cartilage diseases, such as osteoarthritis. Cathepsin K, a cysteine protease, is a stem cell marker used to identify stem cells and the niches they reside in. This research aims to analyse Cathepsin K expression in the zone of Ranvier in 10 day postnatal mice, as well as zebrafish of three ages (3 and 5 days post fertilisation, and adult). Staining through immunohistochemistry, and imaging with fluorescent microscopy were the methods chosen to visualise the expression of each marker as they allow a preservation of the structural integrity of the sample. The images showed a nuclear expression of Cathepsin K in the zone of Ranvier. Meanwhile in zebrafish, it was concentrated in both nuclei and the extracellular matrix of jaw bones. There were no compelling differences in the expression between the developmental stages. The counterstaining performed with known stem cell niche markers helped identify both the location of the stem cell niches where Cathepsin K would be expected and the differences of expression between murine and fish tissues. The findings of this research showed Cathepsin K as one of the markers expressed in the zone of Ranvier, which gives further insight into this stem cell niche and its possible use in the treatment of osteoarthritis and other cartilage degenerative diseases.

The gut microbiota, comprised of a plethora of bacterial species, is a crucial part of the digestive system. The intestinal flora regulates many physiological processes and contributes to a large proportion of an individual's daily energy production. Certain bacteria phyla that reside in the mucus layer of the large intestine play an important role in our physiology. Gut bacteria are capable of processing complex fibres that are present in our diet, influencing organ function, and modulating the immune system. Even slight changes in the microbiota’s composition result in several pathologies. Bacteroides species are known for their ability to metabolize carbohydrates and have acquired interesting mechanisms of processing O- and N-glycosylated proteins, with the latter being the focus of this study. This experiment aims to elucidate the degrading mechanisms of N-glycosylated proteins by Bacteroides species, differentiate between the main degrader species, and analyse the extent of this degradation. Out of all the bacteria strains tested, Bacteroides fragilis and Bacteroides massiliensis displayed proteolytic activity on fetuin, which was further verified through Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and thin-layer chromatography (TLC) analyses. Additionally, six B.fragilis proteins from the GH18 family were cloned and tested for enzymatic activity on various substrates, however, no activity was observed. This work established the phenotypes of bacterial N-glycan degradation.

No description available.

Cancer relapse in myelodysplastic syndrome (MDS) patients receiving bone marrow transplantation can be predicted with measurable residual disease (MRD), by droplet digital polymerase chain reaction (ddPCR). ddPCR quantifies genomic DNA molecules in an absolute manner using end-point amplification. This work aims to demonstrate that ddPCR assay evaluation can be conducted with fewer healthy donor controls compared to methods for relative quantification. The hypothesis is further studied by applying the total error computed in the ddPCR system as a threshold for background noise in personalized assays. Ten assays for detecting MRD markers were evaluated in an optimized PCR-plate setup for accuracy and reproducibility of background in negative controls. Additionally, data analysis of negative controls collected from patient tests complied to the empirical limit of blank based on false - positive counts, in each assay. The findings indicate that the optimized setup accurately determines background noise, and empirical cutoffs for individualized assays are reliable for performance evaluations. This study supports ddPCR integration into clinical settings for personalized mutation analyses in MDS, providing an optimized setup and alternative metrics of evaluating assay performance in respect to the absolute quantification methodology.

Attention deficit hyperactivity disorder (ADHD) is a common neurodevelopmental disorder in children, characterized by symptoms such as hyperactivity, inattention and impulsivity, which often impairs the quality of life and successful performance. The disorder is associated with neurological, genetic and environmental factors affecting brain structure and cognitive function. Treatments include first-line pharmacological treatment, psychological therapy and neurofeedback training. Neurofeedback training with electroencephalography aims to regulate brain activity by targeting the dysregulated theta and beta rhythms, and the theta/beta ratio through personalized or standardized neurofeedback protocols to reduce ADHD symptoms and to improve cognitive function. This study aims to compare differences in the change of theta, beta and the individual alpha peak frequency (iAPF) based on personalized and standardized neurofeedback protocols targeting theta, beta and the theta/beta ratio. Nine children (age 7 to 13) were randomly assigned 16 sessions of personalized or standardized neurofeedback training over a span of two weeks; the neurofeedback protocol was personalized with the help of the iAPF. The theta and beta rhythms and the iAPF were measured by a quantitative electroencephalography and compared before and after the intervention by non-parametric Mann-Whitney-U, Wilcoxon Signed-Rank and Kruskal Wallis tests. These tests showed no significant difference between theta, beta and iAPF before and after the neurofeedback training within and across the personalized and standardized test groups. Previous studies showed an improvement of brain activity and cognitive function through neurofeedback training, but the small sample size and participant artifacts limited this study; concluding that valuable insights into neurofeedback training were gained.

Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders affecting women of reproductive age. The major features of PCOS include polycystic cysts, excess androgens, and polycystic ovaries. Dihydrotestosterone (DHT) is an androgen, and when in excess causes associated with PCOS used to induce PCOS in animal models. In excess, DHT is associated with PCOS features. This study aimed to induce PCOS-like symptoms in Drosophila Melanogaster by supplementing DHT in the diet. The reproductive and metabolic features of PCOS were investigated. Fecundity rates, triglyceride levels, and gene expression levels of Toll and Med genes were assessed. The mean levels of triglycerides (P=0.023) had significantly increased in the high-dose DHT group (F2 generation). The mean expression levels (P=0.02) of the Toll gene were significantly increased in the low-dose DHT group. The treatment of DHT, however, did not affect the fecundity rates of the Drosophila. These findings suggest that DHT induces PCOS-associated metabolic features of lipid alterations and chronic low-grade inflammation. Assessment of the impact on fertility did not give conclusive results thus additional methodologies could be considered in studying the reproductive function. In conclusion, the results obtained in this study highlight the potential of using Drosophila as a model in PCOS research. / <p>Det finns övrigt digitalt material (t.ex. film-, bild- eller ljudfiler) eller modeller/artefakter tillhörande examensarbetet som ska skickas till arkivet.</p><p>There are other digital material (eg film, image or audio files) or models/artifacts that belongs to the thesis and need to be archived.</p>