Plant expression, lyophilisation and storage of HBV medium and large surface antigens for a prototype oral vaccine formulation

Current immunisation programmes against hepatitis B virus (HBV) increasingly often involve novel tri-component vaccines containing—together with the small (S-HBsAg)—also medium and large surface antigens of HBV (M- and L-HBsAg). Plants producing all HBsAg proteins can be a source of components for a potential oral ‘triple’ anti-HBV vaccine. The objective of the presented research was to study the potential of M/L-HBsAg expression in leaf tissue and conditions of its processing for a prototype oral vaccine. Tobacco and lettuce carrying M- or L-HBsAg genes and resistant to the herbicide glufosinate were engineered and integration of the transgenes was verified by PCR and Southern hybridizations. M- and L-HBsAg expression was confirmed by Western blot and assayed by ELISA at the level of micrograms per g of fresh weight. The antigens displayed a common S domain and characteristic domains preS2 and preS1 and were assembled into virus-like particles (VLPs). Leaf tissues containing M- and L-HBsAg were lyophilised to produce a starting material of an orally administered vaccine formula. The antigens were distinctly sensitive to freeze-drying conditions and storage temperature, in the aspect of stability of S and preS domains and formation of multimeric particles. Efficiency of lyophilisation and storage depended also on the initial antigen content in plant tissue, yet M-HBsAg appeared to be approximately 1.5–2 times more stable than L-HBsAg. The results of the study provide indications concerning the preparation of two other constituents, next to S-HBsAg, for a plant-derived prototype oral tri-component vaccine against hepatitis B

K-dron, its mathematical modeling and applications

In this paper we present the definition of K-dron, new geometrical form discovered by Janusz Kapusta in 1985 in New York, its history and connection between geometry and symmetry of a cube. It is worth to emphasize that the authors have derived new formulae for the surface of K-dron using the Laplaceatransform in order to obtain the solution of the boundary-value problem for the partial differential equation describing the vibration of the string. The formula proved by us in this paper is clearer and understandable in view of this structure. The solution obtained in this paper describes in general manner the surface of K-dron and more general surfaces named by us n-K-drons. The formula for the surface of K-dron was derived by the method of Laplacea transform having clear geometrical and physical interpretation because it is presented in linear combination of the equation of planes with the coefficients of directions described by suitable combinations of Heavisides functions. Also wide range and different applications of K-dron are presented.[b]Słowa kluczowe[/b]: K-dron, Laplace’a transform, partial differential of string equation of vibration, initial-boundary value problem

Quality and Pro-Healthy Properties of Belgian Witbier-Style Beers Relative to the Cultivar of Winter Wheat and Raw Materials Used

Unmalted wheat grain and barley malt are the basic materials used in the production of Belgian wheat beers known as Witbier. A change in the ingredients defined in the recipe, by which part of the unmalted wheat is replaced with wheat malt, can positively affect the quality of the beverage produced. The purpose of the study was to brew Witbier-style beers made from four cultivars of winter wheat, with a 50% share of unmalted wheat and barley malt as well as Witbier-style beers made from four wheat cultivars, where 25% of unmalted wheat was replaced with wheat malt. Physicochemical and sensory analyzes showed mild differences in the quality of the beer products, more specifically higher alcohol content (by 11.33%) were found in beers made without the addition of wheat malt, while higher sensory attractiveness and 17.13% higher total polyphenol content were identified in beers enhanced with wheat malt. Phenolic compounds were identified using UPLC-PDA-MS/MS. The highest flavanol content, including kaempferol 3-O-rhamnoside-7-O-pentoside, was found in beers produced using wheat grains of the ‘Elixer’ cultivar, whether or not wheat malt was added; the values were 1.31 mg/L in E50 beer, and 1.39 mg/L in E25 beer. The same beer samples with the highest antioxidant and antiradical activity were found (in E25 beer, 2.35 mmol TE/L, and in E50 beer, 2.12 mmol Fe2+/L). The present findings show that the investigated wheat cultivars may be used in beer production, whereas replacing part of unmalted wheat with wheat malt can improve the sensory profile of the beer produced

Quality and Pro-Healthy Properties of Belgian Witbier-Style Beers Relative to the Cultivar of Winter Wheat and Raw Materials Used

Unmalted wheat grain and barley malt are the basic materials used in the production of Belgian wheat beers known as Witbier. A change in the ingredients defined in the recipe, by which part of the unmalted wheat is replaced with wheat malt, can positively affect the quality of the beverage produced. The purpose of the study was to brew Witbier-style beers made from four cultivars of winter wheat, with a 50% share of unmalted wheat and barley malt as well as Witbier-style beers made from four wheat cultivars, where 25% of unmalted wheat was replaced with wheat malt. Physicochemical and sensory analyzes showed mild differences in the quality of the beer products, more specifically higher alcohol content (by 11.33%) were found in beers made without the addition of wheat malt, while higher sensory attractiveness and 17.13% higher total polyphenol content were identified in beers enhanced with wheat malt. Phenolic compounds were identified using UPLC-PDA-MS/MS. The highest flavanol content, including kaempferol 3-O-rhamnoside-7-O-pentoside, was found in beers produced using wheat grains of the ‘Elixer’ cultivar, whether or not wheat malt was added; the values were 1.31 mg/L in E50 beer, and 1.39 mg/L in E25 beer. The same beer samples with the highest antioxidant and antiradical activity were found (in E25 beer, 2.35 mmol TE/L, and in E50 beer, 2.12 mmol Fe2+/L). The present findings show that the investigated wheat cultivars may be used in beer production, whereas replacing part of unmalted wheat with wheat malt can improve the sensory profile of the beer produced

Organogenesis and long-term micropropagatlon of Polish pea cultivars

The complete protocol for regeneration and long-term micropropagation of several Polish cultivars of pea (Pisum sativum L.) has been elaborated. The shoots were the most likely regenerated via de novo organogenesis. The adventitious buds formed in callus derived from cotyledons tissue adjacent to the axillary meristems of immature embryos. All cultivars' calli regenerated several shoots per explant on the MS medium supplemented with B5 vitamins and 4.5 mgl-1 of BAP, however some differences in regeneration capacity among cultivars were observed. The plantlets were subsequently micropropagated with slightly higher efficiency and preserving a good viability over the long-term culture on a medium containing 2.0 mgl-1 than one with 4.5 mgl-1 of BAP. The additional step of the pre-conditioning culture of multiplicated shoots on a medium with very low BAP concentration i.e. 0.02 mgl-1 was applied and appeared to be beneficial before rooting in vitro or grafting. The modified MS-derived medium with the half-strength of MS macroelements but with the full original dose of calcium and supplemented with B5 vitamins and 1.0 mgl-1 of NAA was developed for effective rooting. The shoots were also sufficiently transferred into ex vitro conditions using grafting. The majority of the regenerated plants had adapted to in vivo conditions in a greenhouse and subsequently has set seeds. The presented protocol provides relatively efficient rate of de novo pea regeneration and would be useful for Agrobacterium-mediated transformation purposes

The Effect of the Addition of Ozonated and Non-Ozonated Fruits of the Saskatoon Berry (<i>Amelanchier alnifolia</i> Nutt.) on the Quality and Pro-Healthy Profile of Craft Wheat Beers

Research into the suitability of domestic raw materials, including, for example, new wheat cultivars and fruit additives for the production of flavoured beers, is increasingly being undertaken by minibreweries and craft breweries. The fruits of the Saskatoon berry are an important source of bioactive compounds, mainly polyphenols, but also macro- and microelements. The fruits of two Canadian cultivars of this species, ‘Honeywood’ and ‘Thiessen’, were used in this study. Physicochemical analysis showed that wheat beers with the addition of non-ozonated fruit were characterised by a higher ethanol content by 7.73% on average. On the other hand, enrichment of the beer product with fruit pulp obtained from the cv. ‘Thiessen’ had a positive effect on the degree of real attenuation and the polyphenol profile. Sensory evaluation of the beer product showed that wheat beers with the addition of ‘Honeywood’ fruit were characterised by the most balanced taste and aroma. On the basis of the conducted research, it can be concluded that fruits of both cvs. ‘Honeywood’ and ‘Thiessen’ can be used in the production of wheat beers, but the fermentation process has to be modified in order to obtain a higher yield of the fruit beer product

Modifying influence of a pesticide dump on plants and animals of wood, meadow, pond and lake ecosystems

The authors realized a Polish national project studying environmental contamination and influence of contamiants on plants and animals (wild mice and fish) around pesticide dump storing about 70 tonns of OCl wastes for over 30 years. The chemical analysis was not very useful because the dump was situated inside the sandy hill (bad sorption) but chnges in plant societies, in contaminats concentrations in animal tissues and, most of all, changes in pathomorphological pattern of animal cells were detected