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      • KCI등재

        Cytotoxicity of Listeriolysin O Produced by Membrane-Encapsulated Bacillus subtilis on Leukemia Cells

        ( Stachowiak,R. ),( L. H. Granicka ),( J. Wisniewski ),( M. Lyzniak ),( J. Kawiak ),( J. Bielecki ) 한국미생물 · 생명공학회 2011 Journal of microbiology and biotechnology Vol.21 No.11

        Encapsulation of biological material in the permiselective membrane allows to construct a system separating cells from their products, which may find biotechnological as well as biomedical applications in biological processes regulation. Application of a permiselective membrane allows avoiding an attack of the implanted microorganisms on the host. Our aim was to evaluate the performance of Bacillus subtilis encapsulated in an elaborate membrane system producing listeriolysin O, a cytolysin from Listeria monocytogenes, with chosen eukaryotic cells for future application in anticancer treatment. The system of encapsulating in membrane live Bacillus subtilis BR1-S secreting listeriolysin O was proven to exert the effective cytotoxic activity on eukaryotic cells. Interestingly, listeriolysin O showed selective cytotoxic activity on eukaryotic cells: more human leukemia Jurkat T cells were killed than human chronic lymphocytic B cells leukemia at similar conditions in vitro. This system of encapsulated B. subtilis, continuously releasing bacterial products, may affect selectively different types of cells and may have future application in local anticancer treatment.

      • THREE-BODY ABRASIVE WEAR IN A BALL-CRATE RING TEST WITH LARGE ABRASIVE PARTICLES

        G.B. STACHOWIAK,G.W. STACHOWIAK 한국트라이볼로지학회 2002 한국트라이볼로지학회 학술대회 Vol.2002 No.10

        Three-body abrasive wear resistance of mild steel, low alloy steel (Bisalloy) and 27%Cr white cast iron was investigated using a ball-cratering test. Glass beads, silica sand, quartz and alumina abrasive particles with sizes larger than 100㎛ were used to make slurries. It was found that the wear rates of all three materials tested increased with time when angular abrasive particles were used and were rather constant when round particles were used. This increase in wear rates was mainly due to the gradual increase in ball surface roughness with testing time. Abrasive particles with higher angularity caused higher ball surface roughness. Mild steel and Bisalloy were more affected by this ball surface roughness changes than the hard white cast iron. Generally, three-body rolling wear dominated. The contribution of two-body grooving wear increased when the ball roughness was significant. More grooves were found when round particles were used or the size of the particles was decreased.

      • KCI등재

        Effect of Illumination Intensities on Astaxanthin Synthesis by Xanthophyllomyces dendrorhous and Its Mutants

        Barbara Stachowiak 한국식품과학회 2013 Food Science and Biotechnology Vol.22 No.4

        The aim of the study was to select optimal illumination for Xanthophyllomyces dendrorhous DSM 5626 and its mutants: 26UV, 10BE, 13B, and 34B for astaxanthin production. The strains were cultivated on YM medium at an illumination of 0-5,000 lx. The highest yields of astaxanthin were found in cultures at 600 lx. The maximum pigment production for the parental strain was 0.95 mg/L and 0.19 g/kg dry cell weight (DCW), while for mutants 1.23-1.51 mg/L and 0.34-0.39 g/kg DCW. It was calculated that the application of adequate values of illumination (within the range of 670-718 lx), determined on the basis of regression models, may improve astaxanthin production by approximately 5-15%, depending on the strain. This was confirmed in the verification experiments for the parental strain and mutants: 10BE, 13B, 26UV, but for 34B at 718 lx the volumetric astaxanthin production was found to be higher than predicted by the model.

      • KCI등재

        Delivery of Chicken Egg Ovalbumin to Dendritic Cells by Listeriolysin O-Secreting Vegetative Bacillus subtilis

        ( Katarzyna Roeske ),( Radoslaw Stachowiak ),( Tomasz Jagielski ),( Michal Kaminski ),( Jacek Bielecki ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.1

        Listeriolysin O (LLO), one of the most immunogenic proteins of Listeria monocytogenes and its main virulence factor, mediates bacterial escape from the phagosome of the infected cell. Thus, its expression in a nonpathogenic bacterial host may enable effective delivery of heterologous antigens to the host cell cytosol and lead to their processing predominantly through the cytosolic MHC class I presentation pathway. The aim of this project was to characterize the delivery of a model antigen, chicken egg ovalbumin (OVA), to the cytosol of dendritic cells by recombinant Bacillus subtilis vegetative cells expressing LLO. Our work indicated that LLO produced by non-sporulating vegetative bacteria was able to support OVA epitope presentation by MHC I molecules on the surface of antigen presenting cells and consequently influence OVA-specific cytotoxic T cell activation. Additionally, it was proven that the genetic context of the epitope sequence is of great importance, as only the native full-sequence OVA fused to the N-terminal fragment of LLO was sufficient for effective epitope delivery and activation of CD8<sup>+</sup> lymphocytes. These results demonstrate the necessity for further verification of the fusion antigen potency of enhancing the MHC I presentation, and they prove that LLO-producing B. subtilis may represent a novel and attractive candidate for a vaccine vector.

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