Faculty of Arts

Předmětem bakalářské práce je zpracování částečné dokumentace pro provádění stavby Fakulty umění Ostravské univerzity. Jako podklad pro práci slouží studie z předmětu Ateliérová tvorba III (urbanistická studie) a Ateliérová tvorba IV (objekt) vypracovaná pod vedením doc. Ing. arch. Josefa Kiszky a dokumentace pro stavební povolení z předmětu Ateliérová tvorba Va pod vedením Ing. Pavla Vlčka, Ph.D. Dokumentace pro provádění stavby byla zpracována dle vyhlášky č. 499/2006 Sb. Úvod textové části se věnuje seznámení s řešeným územím, jeho návazností na související práce a z toho vyplývající řešení projektu, zbytek tvoří průvodní zpráva a souhrnná technická zpráva k objektu. Výkresovou část tvoří výkresová dokumentace, situační výkresy a doklady v požadovaném rozsahu, včetně architektonického detailu.The subject of this thesis is processing of the partial documentation of building Faculty of Arts University of Ostrava. The patterns for this work is the subject Atelier´s work III (urban study) and Atelier´s work IV (object) under the leadership of doc. Ing. Arch. Josef Kiskza and documentation for building permit of the Atelier´s work Va under the leadership of Ing. Pavel Vlček, Ph.D. The documentation of executing the building was processed according to the regulation no. 499/2006 Coll. The first part of the theoretical part is focused on introduction to the solution of the problem, it’s continuity to related works and the resulting solution of this project, the rest is formed by accompanying report and technical summary report of object. Technical drawings consist of technical documentation, situational plans and documentation in requested extent including architectural detail.226 - Katedra architekturyvelmi dobř

Town cemetery with mourning hall, Malacky

Předmětem diplomové práce je rozšíření současného městského hřbitova a návrh nové smuteční síně. Hřbitov je situován na jižním okraji slovenského města Malacky. V současné době se zde nachází pouze plochy pro urnové hroby a tradiční pohřbívání do země. V rámci návrhu jsou tyto plochy rozčleněny a rozšířeny o další typy pohřbívání. Záměrem diplomové práce je urbanistický návrh uspořádání hřbitova a architektonický návrh smuteční síně. Návrh vychází z původní jasně stanovené struktury současného hřbitova a respektování jeho geometrických principů. Účelem je vytvořit klidné místo pro důstojné rozloučení a poslední odpočinek.The subject of this thesis is extension of the present town cemetery and proposal a new mourning hall. The cemetery is situated on the southern outskirts of the slovak city Malacky. It is currently located here only areas for urn graves and traditional burial into the ground. Within the proposal these areas are divided and extended by other types of burials. The main focus of the thesis is urbanism conception of organization of cemetery and architectural proposal of mourning hall. The proposal is based on the original clearly defined structure of the present cemetery and respecting its geometrical principles. The purpose is to create the quiet place for dignified farewell and last rest.226 - Katedra architekturyvelmi dobř

The use of anchored agonists of phagocytic receptors for cancer immunotherapy: B16-F10 murine melanoma model.

The application of the phagocytic receptor agonists in cancer immunotherapy was studied. Agonists (laminarin, molecules with terminal mannose, N-Formyl-methioninyl-leucyl-phenylalanine) were firmly anchored to the tumor cell surface. When particular agonists of phagocytic receptors were used together with LPS (Toll-like receptor agonist), high synergy causing tumour shrinkage and a temporary or permanent disappearance was observed. Methods of anchoring phagocytic receptor agonists (charge interactions, anchoring based on hydrophobic chains, covalent bonds) and various regimes of phagocytic agonist/LPS mixture applications were tested to achieve maximum therapeutic effect. Combinations of mannan/LPS and f-MLF/LPS (hydrophobic anchors) in appropriate (pulse) regimes resulted in an 80% and 60% recovery for mice, respectively. We propose that substantial synergy between agonists of phagocytic and Toll-like receptors (TLR) is based on two events. The TLR ligand induces early and massive inflammatory infiltration of tumors. The effect of this cell infiltrate is directed towards tumor cells, bearing agonists of phagocytic receptors on their surface. The result of these processes was effective killing of tumor cells. This novel approach represents exploitation of innate immunity mechanisms for treating cancer

Interaction of macrophages with melanoma cells labelled with phagocytic ligands. Formation of clusters.

<p>To the mixture of melanoma B16-F10 cells and macrophage cell line PMJ2R laminarin-BAM (A) or free laminarin (B) were added (0.05 mM final concentrations). Photos were taken after 1 hour incubation at 37°C. The experiment was repeated four times with similar results.</p

The effect of anchored ligands of phagocytic receptors on tumor growth and their synergy with LPS.

<p>C57BL/6 mice (females) were inoculated with 4×10⁵ murine melanoma B16-F10 cells per mouse in 0.1 ml RPMI subcutaneously in a shaved area on the right flank. Mice were randomized in groups of 5–6 twelve days after tumor transplantation. Therapies started immediately by intratumoral applications of 50 μl of corresponding solutions and continued every second day for 10 days (together 6 doses). After therapy had commenced, mice were kept individually. Tumors were measured every second day for 14 days and their volume was calculated. (A) Anchored laminarin (laminarin-BAM). Groups of 5 mice obtained 0.2 mM laminarin-BAM in PBS, LPS (0.5 mg/ml PBS), mixture of 0.2 mM laminarin-BAM and LPS (0.5 mg/ml) in PBS, and PBS alone. (B) Anchored mannose. Groups of 6 mice obtained 3 mM mannose-(G)₅₎-(K)₁₀-STE in PBS, LPS (0.5 mg/ml PBS), mixture of 3 mM mannose-(G)₅-(K)₁₀-STE and LPS (0.5 mg/ml) in PBS, and PBS alone. (C) Anchored mannan. Groups of 5 mice obtained 0.2 mM mannan-BAM in PBS, LPS (0.5 mg/ml PBS), mixture of 0.2 mM mannan-BAM and LPS (0.5 mg/ml) in PBS, and PBS alone. (D) Anchored formylpeptide receptor agonist by oligolysin. Groups of 6 mice were injected with 3 mM f-MLF-(G)₅-(K)₁₂ in PBS, LPS (0.5 mg/ml PBS), mixture of 3 mM f-MLF-(G)₅-(K)₁₂ and LPS (0.5 mg/ml) in PBS, and PBS alone. (E) Anchored formylpeptide receptor agonist by stearic acid. The same regime as in (D), 3 mM f-MLF-(G)₅-(K)₁₀-STE used instead of 3 mM f-MLF-(G)₅-(K)_12. *P≤0.05, **P≤0.01, ***P≤0.005, ****P≤0.001 compared to control ■P≤0.05, ■■P≤0.01, ■■■P≤0.005 compared to LPS <b>o</b>P≤0.05, <b>oo</b>P≤0.01, <b>ooo</b>P≤0.005 compared to the ligand.</p

Analysis of cell infiltrate in the tumor during therapy based on the use of f-MLFKK-BAM, LPS and their mixture. Granulocyte detection.

<p>Groups of 9 mice received a single dose of 0.5-MLFKK-BAM, LPS (0.5 mg/ml), mixture of 0.5 mM f-MLFKK-BAM and LPS (0.5 mg/ml), and PBS alone in 50 μl i.t. Preparation of cell suspension and granulocyte staining were performed as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0085222#pone-0085222-g003" target="_blank">Figure 3</a>.</p

Histology.

<p>Melanoma bearing mice were injected i.t. with 50 μl of BAM derivatives of agonists (0.2 mM laminarin-BAM, 0.2 mM mannan-BAM, 0.5 mM f-MLFKK-BAM), their mixtures with LPS (0.5 mg/ml), LPS and PBS alone. Two mice from each group were killed in 24 hours intervals (24, 48, 72 hrs). Excised tumors were fixed with 4% neutral solution of formaldehyde and paraffin blocks were prepared. Sections were stained with hematoxylin/eosin. A– PBS alone; B– effect of particular agonists of phagocytic receptors and LPS alone; C, D– synergistic effect of LPS combinations with particular agonists of phagocytic receptors. Aa – melanoblasts, Ab – necrotic focus with slight granulocyte infiltration, Ba – melanoblasts, Bb – necrotic focus with hemorrhage, Bc – granulation tissue, Bd – slacked edematous ligament, Ca – necrotic tissue with hemorrhage, Cb – negligible residue of tumor, Cc – edematous ligament with inflammatory infiltration, Da – slacked edematous ligament with inflammatory infiltration and hemorrhage foci, Db – bleeding necrosis.</p

Melanoma therapy based on the use of mannan covalently bound to tumor cell surface, synergy with LPS.

<p>Groups of 5 mice were treated starting the 12^th day after tumor transplantation.</p

Analysis of cell infiltrate in the tumor during therapy based on the use of mannan-BAM, LPS and their mixture.

<p>Groups of 9 mice received a single dose of 0.2-BAM in PBS, LPS (0.5 mg/ml PBS), mixture of 0.2 mM mannan-BAM and LPS (0.5 mg/ml) in PBS, and PBS alone in 50 μl i.t. 3 mice from each group were killed in 12, 24 and 48 hours intervals, cells from excised tumors were prepared by enzymatic treatment (Liberase DL and DNase I) and analysed by flow cytometry. The following labelled antibodies were used: (A) anti-Mouse Ly-6G (Gr-1) Alexa Fluor 700 for granulocyte detection, (B) anti-Mouse CD19 APC for detection of B lymphocytes and (C) anti-Mouse NK1.1 PE for NK cells.</p

Synergy of laminarin-BAM with LPS, various regimes of application.

<p>Groups of 4–5 mice were treated starting the 12^th day after tumor transplantation.</p