https://www.um.edu.mt/library/oar/handle/123456789/126924 Tue, 28 Apr 2026 13:47:36 GMT 2026-04-28T13:47:36Z https://www.um.edu.mt/library/oar/handle/123456789/132386 Title: Investigating the viability of cold-stored platelet concentrates and the effects of prolonged incubation Abstract: Thrombocytes, more commonly referred to as platelets, are cellular fragments produced within the bone marrow that play a pivotal role in hemostasis. Cancer patients, trauma patients and individuals who suffer from platelet disorders can benefit from platelet transfusions to ameliorate their condition and state of health. Upon donation, platelet concentrates (PC) are stored at a temperature of 22oC ± 2 oC in gas permeant bags under constant agitation for five days. Currently there are no guidelines available for cold storage and clinical trials are already underway. This study addressed issues related to logistics and blood product wastage by determining whether the quality and viability of platelets are affected if a lower temperature and a longer incubation period is put into practice. In this study, PC were divided into two cohorts, each of which were stored at different temperatures, one in refrigerated conditions (2oC to 6oC) and the other one at room temperature (22oC ± 2 oC). Each aliquot was analysed at different time intervals for parameters which literature has identified as markers of safety and quality. Results were statistically compared to determine whether there was any significant difference between the two cohorts. The investigation yielded predominantly positive results across three main aspects of platelet analysis. Parameters such as the Platelet count (PLT) remained stable, whereas the Mean Platelet Volume (MPV) and the Plateletcrit (PCT) showed significant variations. Platelet metabolism was evident in both room temperature and cold storage, with a notable decreased rate in the latter. pH levels remained within acceptable transfusion thresholds under both storage conditions. No statistically significant differences in cytokine levels were observed between room temperature and cold storage or across different incubation days, although mean cytokine levels were higher at room temperature. In conclusion, an overall positive result was collected for cold-stored platelets however, further studies are still required to fully establish their efficacy in real-life in-vivo scenarios. Description: M.Sc.(Melit.) Mon, 01 Jan 2024 00:00:00 GMT https://www.um.edu.mt/library/oar/handle/123456789/132386 2024-01-01T00:00:00Z https://www.um.edu.mt/library/oar/handle/123456789/131933 Title: Investigation of the stem cell compartment in patients with chronic cytopenias and low-risk MDS Abstract: Myelodysplastic syndromes (MDS) are defined by the WHO as a group of clonal haematopoietic stem cell (HSC) disorders which are characterised by cytopenias, ineffective haematopoiesis, dysplasia, the presence of blasts and an increased risk of developing Acute Myeloid Leukaemia (AML) (WHO,2022) Patients with persistent cytopenias are relatively frequent encounters in routine Haematology screening and the clinical management of these patients can be challenging due to unpredictable clinical course. This study aims to investigate the stem cell compartment of patients with persistent, unexplained cytopenias, and provide insights into the clinical behaviour of these conditions, aiding in better clinical management. In this study, the stem cell compartment of 53 patients was investigated in two separate cohorts: Cohort A (n=30) included patients with persistent cytopenias (potential pre-MDS conditions, or low-risk MDS) and Cohort B (n=23) patients with high-risk MDS and AML, as a control group. A one-tube flow cytometric assay was used for the detection of leukaemic stem cells (LSCs) using a combination of 13 different monoclonal antibodies, to identify immunophenotypic aberrancies. Molecular studies by next-generation sequencing (NGS) were carried out using a targeted myeloid NGS panel to detect any molecular aberrations. Immunophenotypic findings were then correlated with the molecular findings to confirm or otherwise the clonal nature of cytopenias. LSCs were found in 60% of patients from Cohort A and 91% of patients from Cohort B with the most common LSC markers being CD45RA and Combi markers. LSCs were detected at higher percentages in Cohort B. Various molecular aberrations which are commonly associated with MDS and AML were also detected in both Cohorts. There was high agreement between Immunophenotyping and Molecular results in Cohort A (56.6%) and Cohort B (91.3%). Cohort A was further sub-classified into low-risk MDS (50%), ICUS (33%), CCUS (7%) and ‘Other’ (10%) based on cytopenias, dysplasia and the presence of molecular aberrations. The presence of LSCs in 80% of LR-MDS patients, is an important adjunct finding that may prompt clinicians to monitor these patients more closely, with early therapeutic interventions in certain cases. In conclusion, patients with persistent cytopenias together with the presence of LSCs and molecular aberrations might have an increased risk of leukaemic progression and should be monitored more closely. The LSC assay provides valuable information on the stem cell compartment, better guiding clinicians on the course of action for patients with persistent cytopenias. Detection of LSCs is also important in view of the development of therapeutic targets such as immunotherapy targeted towards aberrant markers including CD33, CD123, TIM-3 and CLL-1 leading to more specific and personalised treatments (Hansen et al., 2022). Molecular analysis is also very important for patient stratification, prognosis and targeted therapy. The strong concordance between immunophenotyping and molecular results shows the importance of using a holistic approach when investigating patients with persistent cytopenia and suspected MDS. Description: M.Sc.(Melit.) Mon, 01 Jan 2024 00:00:00 GMT https://www.um.edu.mt/library/oar/handle/123456789/131933 2024-01-01T00:00:00Z https://www.um.edu.mt/library/oar/handle/123456789/127045 Title: TGF-β-induced mesenchymal markers in lung cancer lines Abstract: Lung Cancer is a world-wide issue that has been object of study for years in the research field of molecular pathology. In recent times, it has become more and more important to understand the mechanisms involved in gene-expression and metastasis formation. The Epithelial-to-Mesenchymal transition (EMT) is one of the principal processes involved in metastasis, through a series of molecular changes and signalling pathways. Transforming Growth Factor β (TGF-β) is one of the main inducers of the EMT and has numerous effects on cancer cells behaviour. In this thesis, the gene-expression of mesenchymal and epithelial markers wa investigated in TGF-β-induced lung cancer cell lines. TGF-β was found to have a greater effect on the gene-expression of the A549 and H1975, in contrast to the HCC827 cell line that was found less receptive to the time-dose selected in this study. The gene-expression analysis was carried out using a multiplex approach, the QuantiGene 2.0 Plex Assay that targets several genes involved in the EMT process. An increase in the expression of FN1 that codes for Fibronectin-1 was observed for all the three lung cancer cell lines. A differential expression of other mesenchymal markers was found across the three lung cancer cell lines. These findings confirmed the role of TGF-β on the induction of the EMT process in lung cancer cell lines providing further opportunities to research on the molecular pathways involved in EMT and lung cancer. Description: M.Sc.(Melit.) Mon, 01 Jan 2024 00:00:00 GMT https://www.um.edu.mt/library/oar/handle/123456789/127045 2024-01-01T00:00:00Z