Results support the bipolardiode model of the plasma focus device.

Antibody microarray technology has the potential for playing a large roll in identifying serological biomarker panels for personalized medicine. The aim of this thesis, based on four original papers, was to investigate if information in the serum proteome could be extracted and used for diagnostic, classificational, prognostic or treatment predictive purposes in a range of diseases. In two studies (paper I and paper IV), the diagnosis and prognosis of breast cancer was addressed, also being the main focus of this thesis. In paper I, we identified a biomarker panel capable of stratifying serum samples from metastatic breast cancer patients from those of healthy controls. In paper IV, another panel, pre-validated in the same study, was deciphered that could be used to identify patients destined for metastatic disease in a group of newly diagnosed breast cancer patients. Paper II and III targeted immunotherapy of glioblastoma multiforme and the diagnosis and sub- stratification of two autoimmune diseases (SLE and SSc), respectively. Also in these cases, multiple biomarker panels were identified, each capable of separating predefined cohorts of patients with relevance for applications within personalized medicine. In conclusion, this thesis introduces the concept of personalized medicine; details the antibody microarray technology in general and the platform used for the experiments in paper I to IV; and describes the subsequent microarray data analysis.

04.10.2007 13:24 3kJ Plasma Focus System 4 PhD Thesis.

This thesis presents the thrust modeling of the Dense Plasma Focus plasma motion phases.

X-Rays Emitted from the Plasma Focus,” Ph.D.

The aims of this study have been focused on the identification of cell surface structures involved in induction and regulation of apoptosis. Monoclonal antibodies against the major histocompatibility complex class I (MHC-I) were shown to induce apoptosis when cross-linked on the cell surface of different malignant cells having a pre-B or a myeloid precursor phenotype. The apoptotic response was not epitope dependent, since several different anti-MHC-I antibodies, reacting with different monomorphic determinants of the a chain or b2-microglobulin all induced apoptosis in these cells. However, external cross-linking of antibodies was strictly required for the apoptotic effect. Among cells originating from mature peripheral blood lymphocytes, anti-CD40 stimulated B-cells were susceptible to anti-MHC-I induced apoptosis, whereas resting as well as superantigen activated B- and T- cells were non-responsive to MHC-I ligation. The apoptotic process was not linked to protein kinase C activation or changes in cytoplasmic calcium concentration. In situ terminal doxynucleotidyl transferase staining of apoptotic cells at various stages during MHC-I induced cell death revealed that apoptosis occurred predominantly in the G2/M phase of the cell cycle, with the first apoptotic cells appearing after approximately 12 hours of incubation. The results suggest a role for MHC-I mediated apoptosis during differentiation and activation of certain hematopoietic cells.

THESES - Universal Plasma Focus Laboratory Facility

Molecular recognition is fundamental for the function of biological systems. The properties of the participating biomolecules dictate the type of forces, the strength and the dynamics involved in the interaction. Many interactions are very strong whereas others exhibit weak affinity. Typically, weak interactions work in concert to trigger a biological response. The advantage with this approach is the inherent dynamics. It has been shown that this approach can be successful for in vitro applications as well. By exposing analytes to a multitude of specific, weak affinity interactions, which are governed by fast association and dissociation rates, separation based on small differences in affinity is possible. The same principles can also be used to characterize biological weak affinity binders and for analytical purposes. This thesis has discussed biomolecular interactions in the weak affinity range (defined in this investigation as dissociation constants (KD) larger than 10 mM) in general and has focused on how they can be studied and exploited in vitro. Weak affinity monoclonal antibodies were used as model systems in three different applications: (i) to explore how weak affinity chromatography based on monoclonal IgM can be employed to separate structurally related steroids under non-denaturing conditions. (ii) to investigate the possibilities of studying weak interactions between antibodies and haptens with an optical biosensor based on surface plasmon resonance. A rationale for the design of such experiments to avoid inaccurate results was suggested. (iii) to introduce continuous real-time immunosensing for monitoring fluctuating concentrations of biomolecules in a flow.

As a radiation source the plasma focus PF is operated in time-matched regime TMR.

Development and studies of a small plasma focus

Dendritic cells (DC) are a heterogeneous lineage of bone-marrow-derived leukocytes that serve as the link between innate and adaptive immunity. They are professional antigen-presenting cells that play an important protective and regulatory role in both health and disease. This thesis is based upon six original papers that deal with the function and transcription of human DCs, with focus on their role in the inflammatory immune responses, such as allergic rhinitis. Transcriptional profiling of DCs with microarrays has been an extensively utilized technique in the projects presented in this thesis. We have evaluated the gene expression profiles and functions of two in vitro models of human DCs, namely the monocyte-derived DCs and the differentiated cell line MUTZ-3, and the transcriptional regulation induced in these models by pro-inflammatory signals. We have also studied the effect of allergenic stimulation on the transcription and function of DCs derived from healthy and allergic individuals. Two allergens were used, the detergent enzyme lipase and grass pollen, to evaluate the direct effect on phenotype and gene expression of DCs in addition to the subsequent ability of allergen-challenged DCs to amplify and modify the autologous effector T cell response. We demonstrate that the transcriptional responses of DCs and effector T cells to allergenic stimulation are different between allergic and healthy individuals. These transcriptional profiles involved in the immune recognition of allergens will be further evaluated in order to understand the interplay between DCs and T cells in allergic rhinitis. Furthermore, in addition to the in vitro models studied, we have performed phenotypical and transcriptional characterizations of in vivo DCs isolated from peripheral blood and tonsillar tissue. We suggest that follicular DCs in tonsils may have previously unacknowledged costimulatory functions in the germinal center reaction, as they express CD137. An extensive transcriptional profiling of freshly sorted DC subsets from blood and tonsils identified DC-subset selective gene expression and pinpoint their relationships. We demonstrate innate specialization of these subsets and show that the environment in tonsils determines the transcriptional activity of myeloid DCs. In conclusion, these studies have provided insight in the transcription and phenotype of in vivo immature/mature DC populations as well as in the immune response induced by allergens or inflammatory signals.

Plasma Focus Rawat 12 | Plasma (Physics) | Capacitor

B cell lymphomas (BCLs) are a group of severe cancers, afflicting both men and women at different ages. Gene expression profiling (GEP), have during the latest decades fundamentally contributed to a better classification and biological understanding of the different BCLs. Most studies using GEP have analyzed mRNA from tumor tissue constituted of both tumor cells and non-malignant bystander cells. This thesis is based on four original papers where pure, cell-sorted BCL cells are analyzed by GEP. This procedure allows identification of differences in tumor cells rather than variations due to cellular composition. These studies focus on identifying new prognostic, diagnostic and/or functional markers for BCLs, using four different approaches. In the first study, GEP of pure mantle cell lymphoma (MCL) cells were used to identify MCL-associated targets genes. Subsequent immunohistochemistry (IHC) using antibodies raised against a unique protein epitope signature (PrEST) was performed. Using this high-throughput strategy we were able to identify proteins either uniquely or highly expressed in MCL compared to normal lymphoid tissue. This study confirmed the usage of transcriptional screening to identify molecular tumor-associated targets, as well as suggest the PrEST-approach as a novel efficient technique to identify molecular targets with both a known and unknown identity. The indolent growing follicular lymphoma (FL) cells are dependent on interplay with cells in the microenvironment for survival. We analyzed, in a second study, pure FL cells for highly expressed genes encoding surface bound proteins. An aberrant expression of CX3CR1 on the surface, not only on FL cells but also on tumor cells in several other BCLs was identified. CX3CR1 is suggested to be involved in site-specific dissemination, the possibility to use CX3CR1 as a target for antibody-mediated intervention needs however further investigations. The indolent FL often transforms to diffuse large B-cell lymphoma (DLBCL-tr). Previous studies analyzing tumor tissue for markers involved in this high-grade transformation have revealed results with low concordance. In a, we analyzed purified FL and DLBCL-tr tumor cells and found that large inter-tumor heterogeneity still was observed. Despite the heterogenous gene expression, this third study identified distinct differences between tumor entities rather than differences due to various composition of bystander cell and two proteins, galectin-3 and NEK2 pinpointing a subgroup of DLBCL-tr over FL were identified. In the last study, we used a unique material of eight different B cell lymphomas and identified for the first time unique transcription factors for each of these highly purified entities. The identified TFs are not only potential new molecular targets but are also partly responsible for the differences in global gene expression in the different BCLs as demonstrated by unsupervised clustering. In conclusion, analyzing pure lymphoma cells, dramatically enhance the resolution of analyses and allowed for identification of tumor-cell related genes using different approaches. Downstream analyzes of these genes allow for identification of proteins with a major biological importance for BCLs and potentially of new biomarkers.

This thesis will focus on nanosecond pulsed DBDs.

In this thesis we have addressed technical challenges such as optimising proteomic coverage by evaluating different protein and peptide separation methods and mapping proteins to organelles with density gradient organelle fractionation followed by proteomic quantification of the fractions. Further, we applied both a discovery mode of proteomics and a focussed pathway centric method to evaluate the response of breast cancer cell lines to chemotherapeutic agents. Proteomic pathway analysis was also utilised to investigate and explain the differential response to therapy in different breast tumour subpopulations.