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The green capsule: edible vaccine production in transgenic plants
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Abstracts / New Biotechnology 33S (2016) S1–S213
regulate host defenses against pathogens, lipid metabolism, and developmental events. Development of diverse steryl-a-glycosides may be highly useful for identifying potential cholesterol aglucosyltransferase inhibitor candidates and therapeutic target to prevent H. pyroli-induced peptic ulcer, gastric carcinoma, and mucosal associated lymphoid tissue lymphoma. In this study, we report the acceptor diversity of HP0421 from H. pyroli based on its specificity for structurally related sterol or steroid substrates for a-glycosylation. Enzymatic glucosylation of HP0421 from H. pyroli with 24 sterols or steroids show protein plasticity on substrate specificity, generating 10 steryl-a-glucosides, in addition to reported cholesteryl-a-glucoside. Investigation of the effect of trans-androsterone-a-glucoside on tamoxifen-treated MCF-7 breast cancer cells shows dose-dependent depression of cell viability and enhanced drug effectiveness, indicating the potential pharmaceutical applications of steryl-a-glucosides. http://dx.doi.org/10.1016/j.nbt.2016.06.979
P3-11 The green capsule: edible vaccine production in transgenic plants Helga Zelenyánszki ∗ , Zoltán Mezei, Éva Hamar, Gábor Tóth, László Tamás Eötvös Loránd University, Hungary Grains of transgenic cereals are suitable for the production, storage and delivery of pharmaceutical proteins like edible vaccines. Proteins expressed in the endosperm can retain biological activity for years at room temperature due to dehydration occurred during grain maturation and desiccation. Abundance of chaperons and disulphide-isomerases in the endosperm also promote stability of the proteins. To induce proper mucosal immune response adjuvants and/or carrier molecules are needed due to the tolerogenic nature of mucosal immunity. CTB, the non-toxic B subunit of the Vibrio cholerae toxin is strong mucosal adjuvant. The final goal of our work is to establish transgenic barley lines expressing a fusion protein comprising flagellin (FliC) of Salmonella Enteritidis and CTB in a strictly endosperm specific manner. The grains of this transgenic barley could be used for mass immunization of avian against Salmonella Enteritidis readily and at low cost. Characterisation of the immunological properties of the recombinant fusion protein is going to be carried out on transgenic Arabidopsis thaliana plants. Two transformation cassettes were assembled with different promoters. The ubiquitin-1 promoter of maize is responsible for constitutive expression of the gene of interest in barley and also in A. thaliana. The HMW-glutenin promoter of wheat drives strong tissue specific expression in the barley endosperm, and also in the seeds of Arabidopsis. Agrobacterium mediated plant transformation was carried out with both gene constructs using the floral dip method on Arabidopsis. Characterisation of the T1 generation on DNA, RNA and protein levels is in progress. http://dx.doi.org/10.1016/j.nbt.2016.06.980
P3-12 Pentoxifylline affects idarubicin binding to DNA Grzegorz Golunski ∗ , Agnieszka Borowik, Andrea Lipinska, ´ Monika ´ Romanik, Natalia Derewonko, Anna Woziwodzka, Jacek Piosik Intercollegiate Faculty of Biotechnology University of Gdansk & Medical University of Gdansk, Poland Anthracycline anticancer drugs are commonly used in treatment of various types of leukemia, carcinoma and sarcoma. The most prominent representative of this group is doxorubicin (DOX)–compound extensively studied for its biological properties, anticancer activity and as model anticancer drug in chemotherapy modifications studies. However, idarubicin (IDA) – DOX derivative – is not as broadly studied, with number of published reports 28 times lower than for DOX. IDA biophysical properties are not well established and its potential direct interactions with other biologically active aromatic compounds, such as pentoxifylline (PTX) – methylxanthines representative – were not studied at all. Numerous reports present potential sequestration of aromatic ligands by other biologically active aromatic compounds. One of the representatives of this group is PTX, which is a perfect candidate for modulator of the anticancer drugs, as PTX possesses anticancer activity itself. It is suggested that potential interactions of anticancer drugs with PTX may reduce side effects of the chemotherapy without affecting anticancer activity of the drugs. In this work we analyzed IDA biophysical properties and assessed influence of PTX on IDA interactions with DNA. To achieve this goal we employed spectrophotometric methods coupled with analysis with the appropriate mathematical models as well as flow cytometry and Ames test. Obtained results show PTX influence on IDA binding to DNA and are well in agreement with the data previously published for other aromatic ligands. Additionally it may be hypothesized that direct interactions between IDA and PTX may influence the anticancer drug biological activity. http://dx.doi.org/10.1016/j.nbt.2016.06.981
P3-13 Development of small molecule inhibitors of IL-1 processing Piotr Kowalczyk ∗ , Stefan Chmielewski, Aleksandra Poczkaj, ´ Magdalena Salwinska, Aniela Gołas, Karolina Gluza, Michał ˛ Gałezowski, Oleksandr Levenets, Jakub Woyciechowski, Marta Bugaj, Charles Fabritius, Krzysztof Brzózka Selvita S.A., Poland Release of the cytokines from the IL-1 family is crucial for the development of innate immune responses and subsequent stimulation and modulation of the adaptive immunity. Due to its strong pro-inflammatory activity, the production of this cytokines is very tightly controlled. Excessive production of the cytokines from the IL-1 family e.g. IL-1 is a hallmark of many autoinflammatory and autoimmune disorders including rheumatoid arthritis or Crohn’s disease. Under physiological condition, IL1  is produced at very low level but upon stimulation of wide range of endogenous and exogenous factors its expression is rapidly induced at both transcriptional and translational level. Such stimulation results also in the formation of multiprotein complex known as inflammasome. Active inflammasome forms multiprotein platform that specifically activates caspase-1 which in turn facilitates proteolytic activation and releases of the active form of IL-1. In this study, we report development of IL-1 processing inhibitors. Compounds,
Abstracts / New Biotechnology 33S (2016) S1–S213
regulate host defenses against pathogens, lipid metabolism, and developmental events. Development of diverse steryl-a-glycosides may be highly useful for identifying potential cholesterol aglucosyltransferase inhibitor candidates and therapeutic target to prevent H. pyroli-induced peptic ulcer, gastric carcinoma, and mucosal associated lymphoid tissue lymphoma. In this study, we report the acceptor diversity of HP0421 from H. pyroli based on its specificity for structurally related sterol or steroid substrates for a-glycosylation. Enzymatic glucosylation of HP0421 from H. pyroli with 24 sterols or steroids show protein plasticity on substrate specificity, generating 10 steryl-a-glucosides, in addition to reported cholesteryl-a-glucoside. Investigation of the effect of trans-androsterone-a-glucoside on tamoxifen-treated MCF-7 breast cancer cells shows dose-dependent depression of cell viability and enhanced drug effectiveness, indicating the potential pharmaceutical applications of steryl-a-glucosides. http://dx.doi.org/10.1016/j.nbt.2016.06.979
P3-11 The green capsule: edible vaccine production in transgenic plants Helga Zelenyánszki ∗ , Zoltán Mezei, Éva Hamar, Gábor Tóth, László Tamás Eötvös Loránd University, Hungary Grains of transgenic cereals are suitable for the production, storage and delivery of pharmaceutical proteins like edible vaccines. Proteins expressed in the endosperm can retain biological activity for years at room temperature due to dehydration occurred during grain maturation and desiccation. Abundance of chaperons and disulphide-isomerases in the endosperm also promote stability of the proteins. To induce proper mucosal immune response adjuvants and/or carrier molecules are needed due to the tolerogenic nature of mucosal immunity. CTB, the non-toxic B subunit of the Vibrio cholerae toxin is strong mucosal adjuvant. The final goal of our work is to establish transgenic barley lines expressing a fusion protein comprising flagellin (FliC) of Salmonella Enteritidis and CTB in a strictly endosperm specific manner. The grains of this transgenic barley could be used for mass immunization of avian against Salmonella Enteritidis readily and at low cost. Characterisation of the immunological properties of the recombinant fusion protein is going to be carried out on transgenic Arabidopsis thaliana plants. Two transformation cassettes were assembled with different promoters. The ubiquitin-1 promoter of maize is responsible for constitutive expression of the gene of interest in barley and also in A. thaliana. The HMW-glutenin promoter of wheat drives strong tissue specific expression in the barley endosperm, and also in the seeds of Arabidopsis. Agrobacterium mediated plant transformation was carried out with both gene constructs using the floral dip method on Arabidopsis. Characterisation of the T1 generation on DNA, RNA and protein levels is in progress. http://dx.doi.org/10.1016/j.nbt.2016.06.980
P3-12 Pentoxifylline affects idarubicin binding to DNA Grzegorz Golunski ∗ , Agnieszka Borowik, Andrea Lipinska, ´ Monika ´ Romanik, Natalia Derewonko, Anna Woziwodzka, Jacek Piosik Intercollegiate Faculty of Biotechnology University of Gdansk & Medical University of Gdansk, Poland Anthracycline anticancer drugs are commonly used in treatment of various types of leukemia, carcinoma and sarcoma. The most prominent representative of this group is doxorubicin (DOX)–compound extensively studied for its biological properties, anticancer activity and as model anticancer drug in chemotherapy modifications studies. However, idarubicin (IDA) – DOX derivative – is not as broadly studied, with number of published reports 28 times lower than for DOX. IDA biophysical properties are not well established and its potential direct interactions with other biologically active aromatic compounds, such as pentoxifylline (PTX) – methylxanthines representative – were not studied at all. Numerous reports present potential sequestration of aromatic ligands by other biologically active aromatic compounds. One of the representatives of this group is PTX, which is a perfect candidate for modulator of the anticancer drugs, as PTX possesses anticancer activity itself. It is suggested that potential interactions of anticancer drugs with PTX may reduce side effects of the chemotherapy without affecting anticancer activity of the drugs. In this work we analyzed IDA biophysical properties and assessed influence of PTX on IDA interactions with DNA. To achieve this goal we employed spectrophotometric methods coupled with analysis with the appropriate mathematical models as well as flow cytometry and Ames test. Obtained results show PTX influence on IDA binding to DNA and are well in agreement with the data previously published for other aromatic ligands. Additionally it may be hypothesized that direct interactions between IDA and PTX may influence the anticancer drug biological activity. http://dx.doi.org/10.1016/j.nbt.2016.06.981
P3-13 Development of small molecule inhibitors of IL-1 processing Piotr Kowalczyk ∗ , Stefan Chmielewski, Aleksandra Poczkaj, ´ Magdalena Salwinska, Aniela Gołas, Karolina Gluza, Michał ˛ Gałezowski, Oleksandr Levenets, Jakub Woyciechowski, Marta Bugaj, Charles Fabritius, Krzysztof Brzózka Selvita S.A., Poland Release of the cytokines from the IL-1 family is crucial for the development of innate immune responses and subsequent stimulation and modulation of the adaptive immunity. Due to its strong pro-inflammatory activity, the production of this cytokines is very tightly controlled. Excessive production of the cytokines from the IL-1 family e.g. IL-1 is a hallmark of many autoinflammatory and autoimmune disorders including rheumatoid arthritis or Crohn’s disease. Under physiological condition, IL1  is produced at very low level but upon stimulation of wide range of endogenous and exogenous factors its expression is rapidly induced at both transcriptional and translational level. Such stimulation results also in the formation of multiprotein complex known as inflammasome. Active inflammasome forms multiprotein platform that specifically activates caspase-1 which in turn facilitates proteolytic activation and releases of the active form of IL-1. In this study, we report development of IL-1 processing inhibitors. Compounds,