Metabolic and bioprocess engineering for production of selenized yeast with increased content of seleno-methylselenocysteine.

Specific Se-metabolites have been recognized to be the main elements responsible for beneficial effects of Se-enriched diet, and Se-methylselenocysteine (SeMCys) is thought to be among the most effective ones. Here we show that an engineered Saccharomyces cerevisiae strain, expressing a codon optimized heterologous selenocysteine methyltransferase and endowed with high intracellular levels of S-adenosyl-methionine, was able to accumulate SeMCys at levels higher than commercial selenized yeasts. A fine tuned carbon- and sulfate-limited fed-batch bioprocess was crucial to achieve good yields of biomass and SeMCys. Through the coupling of metabolic and bioprocess engineering we achieved a ∼24-fold increase in SeMCys, compared to certified reference material of selenized yeast. In addition, we investigated the interplay between sulfur and selenium metabolism and the possibility that redox imbalance occurred along with intracellular accumulation of Se. Collectively, our data show how the combination of metabolic and bioprocess engineering can be used for the production of selenized yeast enriched with beneficial Se-metabolites.

Arsenosugars and other arsenic compounds in littoral zone algae from the Adriatic Sea.

In 10 different marine algae from the littoral zone (found between the highest and lowest tide marks on the seashore) arsenic compounds were determined by means of a high-performance liquid chromatography (anion and cation exchange)-UV photochemical digestion-hydride generation-atomic fluorescence spectrometry (HPLC-UV-HGAFS) system. Samples (Ceramium sp., Cystoseira barbata, Enteromorpha sp., Fucus virsoides, two different species of Gelidium, Padina pavonica, Polisyphonia sp. and Ulva rigida) were collected along the Adriatic Sea coast of Slovenia. The total arsenic content of the algal samples, as determined by ICP-MS, ranged from 1.35 to 28.1 microg g(-1) (fresh weight). In all algae but two, the most abundant arsenic species found were arsenosugars with minor amounts of other arsenic compounds. Cystoseira barbata and Ceramium sp. contained high amounts of mainly inorganic arsenic. A small quantity of arsenobetaine was detected in most of the investigated Adriatic algae, which probably originates from mesofauna attached to the algae in their natural habitat.

Selenium species determination in selenium-enriched pumpkin (Cucurbita pepo L.) seeds by HPLC-UV-HG-AFS.

Pumpkins were treated by spraying the leaves in the flowering period with a water solution containing 1.5 mg Se per liter in the form of Na2SeO4. The average total selenium content of seeds was found to be 0.19 microg g(-1) in nontreated pumpkins and 1.1 microg g(-1) in exposed ones. For speciation analysis, enzymatic hydrolysis with different amounts of Protease XIV was carried out. Under optimal conditions of enzymatic hydrolysis, 90% of the total selenium was found in soluble forms. Separation of species was performed using HPLC on anion and cation exchange columns and for detection UVHG-AFS was applied. In enzymatic hydrolysis extracts, the main fraction of selenium was bound as selenomethionine (SeMet), representing on average of 81 +/- 8% of the total Se content in the sample.

Consumption of organic diets does not affect intake and absorption of zinc and copper in men - evidence from two cross-over trials.

Agricultural methods may affect the nutritional composition of plants and cause complex changes in the food matrix. Whether this affects the dietary absorption of minerals that are important for maintaining health thorough life remains unclear. We compared the effects of organic and conventional diets on intake and absorption of zinc and copper in men. Two double-blinded, cross-over, intervention trials (3 dietary periods of 12 days with 2-week-long wash-out) were performed in 2008 (n = 17) and 2009 (n = 16) in young men. The diets were based on 9 crops grown in rigidly controlled organic and conventional systems in 2 replications over 2 years. The primary outcomes were intake and absorption of zinc and copper. The absorption was determined by faecal excretion of stable enriched isotopes extrinsically added to the entire menu. Within each year, the intake and absorption of zinc (overall mean ± SD; 12.35 ± 0.47 mg per 10 MJ and 44.6% ± 12.1, respectively) and copper (overall mean ± SD; 2.12 ± 0.28 mg per 10 MJ and 41.2% ± 13.2, respectively) were not different between the organic and conventional diets. The growing season had no effect on zinc intake and absorption, but the copper intake was higher (P = 0.01) and absorption lower (P < 0.005) in 2008 compared with 2009 (overall mean absorption ± SD; 35.3% ± 13.5 in 2008 and 54.0% ± 10.7 in 2009). In conclusion, organic agriculture does not affect the intake and absorption of copper and zinc in men. Consequently, it does not seem to promote the health beneficial role of these minerals in vivo.

In vitro and in vivo characterization of meloxicam nanoparticles designed for nasal administration.

The nasal pathway represents a non-invasive route for delivery of drugs to the systemic circulation. Nanonization of poorly soluble drugs offers a possibility to increase dissolution properties, epithelial permeability or even bioavailability. The aim of the present study was to use in vitro methods to screen formulations which were intended for nasal application, and to perform animal experiments for recognizing the differences in plasmakinetics of intranasal- and oral-administered meloxicam nanoparticles. Due to nanonization the solubility of meloxicam elevated up to 1.2mg/mL, additionally the extent of dissolution also increased, complete dissolution was observed in 15 min. Favorable in vitro diffusion profile of meloxicam nanoparticles was observed and their epithelial permeability through human RPMI2650 cells was elevated. The pharmacokinetic parameters were significantly increased when meloxicam was administered as nanoparticles to rats either nasally (increase of Cmax 2.7-fold, AUC 1.5-fold) or orally (increase of C(max) 2.4-fold, AUC 2-fold) as compared to physical mixture of the drug and the excipients.

Functional food. Product development, marketing and consumer acceptance--a review.

It was mainly the advances in understanding the relationship between nutrition and health that resulted in the development of the concept of functional foods, which means a practical and new approach to achieve optimal health status by promoting the state of well-being and possibly reducing the risk of disease. Functional foods are found virtually in all food categories, however products are not homogeneously scattered over all segments of the growing market. The development and commerce of these products is rather complex, expensive and risky, as special requirements should be answered. Besides potential technological obstacles, legislative aspects, as well as consumer demands need to be taken into consideration when developing functional food. In particular, consumer acceptance has been recognized as a key factor to successfully negotiate market opportunities. This paper offers a brief overview of the current functional food market situation in USA, Japan and some European countries completed with some comments on functional food future potential. It explores the main challenges of such product development focusing on the different factors determining the acceptance of functional food. Furthermore it discusses some prominent types of these food products currently on the market.

Bioavailability of selenium from selenium-enriched green onions (Allium fistulosum) and chives (Allium schoenoprasum) after 'in vitro' gastrointestinal digestion.

Three sample preparation methods--proteolysis to determine the initial species distribution, and an in vitro gastric and gastrointestinal digestion to assess the bioavailability of selenium--were applied to extract the selenium from selenized green onion and chive samples. Ion exchange chromatography was coupled to a high-performance liquid chromatography-ICP-MS system to analyze the selenium species of Allium samples. The difference in the selenium accumulation capability of green onions and chives was significant. Chive accumulated a one order of magnitude higher amount of selenium than did green onion. After proteolysis of both types of Allium plants, high amounts of organic selenium species such as MeSeCys, SeCys2 and SeMet became accessible. In the case of Se(VI)-enrichment, selenate was the main species in the proteolytic extract. After simulating the human digestion, the organic species were just slightly bioavailable compared with the results from proteolysis. The inorganic selenium content of the selenized samples increased significantly and SeOMet could be detected from the extracts. As an effect of the significant pH change between the gastric and the intestinal tracts, two oxidation processes took place: selenite oxidized to selenate, while SeMet oxidized to SeOMet.