Phage-Derived Fully Human Monoclonal Antibody Fragments to Human Vascular Endothelial Growth Factor-C Block Its Interaction with VEGF Receptor-2 and 3

Vascular endothelial growth factor C (VEGF-C) is a key mediator of lymphangiogenesis, acting via its receptors VEGF-R2 and VEGF-R3. High expression of VEGF-C in tumors correlates with increased lymphatic vessel density, lymphatic vessel invasion, sentinel lymph node metastasis and poor prognosis. Recently, we found that in a chemically induced skin carcinoma model, increased VEGF-C drainage from the tumor enhanced lymphangiogenesis in the sentinel lymph node and facilitated metastatic spread of cancer cells via the lymphatics. Hence, interference with the VEGF-C/VEGF-R3 axis holds promise to block metastatic spread, as recently shown by use of a neutralizing anti-VEGF-R3 antibody and a soluble VEGF-R3 (VEGF-C/D trap). By antibody phage-display, we have developed a human monoclonal antibody fragment (single-chain Fragment variable, scFv) that binds with high specificity and affinity to the fully processed mature form of human VEGF-C. The scFv binds to an epitope on VEGF-C that is important for receptor binding, since binding of the scFv to VEGF-C dose-dependently inhibits the binding of VEGF-C to VEGF-R2 and VEGF-R3 as shown by BIAcore and ELISA analyses. Interestingly, the variable heavy domain (VH) of the anti-VEGF-C scFv, which contains a mutation typical for camelid heavy chain-only antibodies, is sufficient for binding VEGF-C. This reduced the size of the potentially VEGF-C-blocking antibody fragment to only 14.6 kDa. Anti-VEGF-C VH-based immunoproteins hold promise to block the lymphangiogenic activity of VEGF-C, which would present a significant advance in inhibiting lymphatic-based metastatic spread of certain cancer types

Self-supporting oven switch contoler product development project

Predmet diplomske naloge je razvoj novega izdelka z metodami in tehnikami menedžmenta vrednosti. Metodologijo smo praktično uporabili v podjetju Eurel, pri izdelku »Samonosilni stikalni gumb pečice«, ki je krmilni gumb pečic in kuhalnih plošč. Izdelek je razvit na pobudo podjetja Gorenje, na voljo pa bo tudi drugim branžnim proizvajalcem. V praktičnem delu diplomske naloge sem prikazal rabo tehnik vrednostne analize v procesu razvoja novega izdelka. Najprej smo definirali funkcije izdelka, nato delne rešitve funkcij, s kombinacijo teh pa več predlogov izvedbe izdelka. Predloge smo nato ovrednotili in izbrali najboljši predlog izdelka. Nazadnje sem izdelal še stroškovno kalkulacijo, določil prodajno ceno izdelka, projektni plan in naredil pripravo za zagon izvajanja projekta razvoja izdelka v podjetju Eurel.In this diploma we present the process of new product development, with use of value management methods and techniques. Practical use of value management was applied in product development Selfsupporting owen switch controller, which was developed on an initiative by company Gorenje. In practical part of diploma, we applied value analysis techniques to the product development porces, that includes definition of product functions, then performing value analysis and selecting the best proposed product variant. Finnaly we performed cost calculation and product economics, we defined own and selling price, we prepared project plan and product development start.up project

Cognitive impairments in patients with the Parkinsons disease

As the motor disturbances are the most well known, easily observed and the only ones used in the diagnosis of the Parkinsons disease (PD), the non-motor features of the PD are often overlooked even though the cognitive impairments in the area of direction and sustenance of attention, working memory and other executive functions present a most disabling effect on the ability to cope with everyday life tasks. There is no reason to assume that the degeneration in PD and its effect are limited to motor loop only, but could affect the rest of the cortico-subcortico-cortical loops as well, resulting in specific cognitive dysfunctions related to individual "cognitive" loops. Specifically the impairment of dorsolateral loop should lead to dysfunction of working memory and other executive abilities, the impairment of orbitofrontal loop typically results in lack of social control, obsessive and compulsive behavior, emotional instability and impulsiveness, while anterior cingulate loop is related to dysfunctions of attention and consequent decreased motivation and lack of interest

MyoD forms micelles which can dissociate to form heterodimers with E47: implications of micellization on function.

MyoD is a member of a family of DNA-binding transcription factors that contain a helix-loop-helix (HLH) region involved in protein-protein interactions. In addition to self-association and DNA binding, MyoD associates with a number of other HLH-containing proteins, thereby modulating the strength and specificity of its DNA binding. Here, we examine the interactions of full-length MyoD with itself and with an HLH-containing peptide portion of an E2A gene product, E47-96. Analytical ultracentrifugation reveals that MyoD forms micelles that contain more than 100 monomers and are asymmetric and stable up to 36 degrees C. The critical micelle concentration increases slightly with temperature, but micelle size is unaffected. The micelles are in reversible equilibrium with monomer. Addition of E47-96 results in the stoichiometric formation of stable MyoD-E47-96 heterodimers and the depletion of micelles. Micelle formation effectively holds the concentration of free MyoD constant and equal to the critical micelle concentration. In the presence of micelles, the extent of all interactions involving free MyoD is independent of the total MyoD concentration and independent of one another. For DNA binding, the apparent relative specificity for different sites can be affected. In general, heterodimer-associated activities will depend on the self-association behavior of the partner protein

Long-term Cognitive Deficits in Patients with Good Outcomes after Aneurysmal Subarachnoid Hemorrhage from Anterior Communicating Artery

Aim: To evaluate long-term cognitive consequences of subarachnoid hemorrhage with good outcome and the opinion of patients and their relatives about these consequences. Methods: The study included 10 patients surgically treated for subarachnoid hemorrhage due to the rupture of aneurysm of the anterior communicating artery 2 or more years earlier, and 10 age- and sex-matched healthy controls. The preoperative and postoperative course in the patients was uneventful. Clinical and psychosocial factors and cognitive status of the patients were assessed by use of checklists and neuropsychological tests for executive functions, attention, and memory, and event-related potential recordings (waves P3a and P3b) with tree-stimulus auditory oddball paradigm, which was also performed in healthy controls. Results: The number of reported cognitive problems negatively correlated with the patients’ level of community integration (ρ range, -0.22 to -0.75). The average neuropsychological results ranged between the 12th and 46th percentile. Impaired results were found in 7 patients across different tests and were most frequent for visual memory, followed by verbal memory and executive functions. A clear decline in cognitive functioning was observed in 3 patients. Neither P3a nor P3b wave could be found in 3 patients. In comparison with controls, patients had significantly longer P3b wave latencies (364 vs 334 ms; Mann-Whitney U test, P = 0.025). We found statistically non-significant, but still prominent negative correlations between the sustained attention results and latencies of P3a (ρ = -0.58; P = 0.172) and P3b (ρ = -0.58; P = 0.172) waves. Conclusion: Despite good outcome after subarachnoid hemorrhage, persistent cognitive consequences were still manifest, limiting the patients\u27 psychosocial functioning. The correlation between neuropsychological and neurophysiological measures indicated frontal lobe damage, which in some patients persisted for years after the hemorrhage