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.

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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.

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