A pot experiment, under greenhouse conditions, was carried out aiming at investigating the agronomic biofortification of alfalfa (Medicago sativa L.) with Se and monitoring the Se uptake and accumulation dynamics within four consecutive harvests within the same growing season. Two ionic Se forms, i.e., sodium selenate (Se (VI)) and sodium selenite (Se (IV)), were applied once at a rate of 1, 10, and 50 mg kg⁻¹ (added on Se basis), while 10 and 50 mg L⁻¹ of a red elemental Se (red Se⁰) were used; all Se treatments were added as soil application. Application of Se (VI) at the rate of 50 mg kg⁻¹ was toxic to alfalfa plants. The effect of Se forms on Se accumulation in alfalfa tissues, regardless of the applied Se concentration, follows: Se (VI) > Se (IV) > red Se⁰. The leaf, in general, possessed higher total Se content than the stem in all the treatments. The accumulation of Se in stem and leaf tissues showed a gradual decline between the harvests, especially for plants treated with either Se (VI) or Se (IV); however, the chemically synthesized red Se⁰ showed different results. The treatment of 10 mg kg⁻¹ Se (VI) resulted in the highest total Se content in stem (202.5 and 98.0 µg g⁻¹) and leaf (643.4 and 284.5 µg g⁻¹) in the 1st and 2nd harvests, respectively. Similar tendency is reported for the Se (IV)-treated plants. Otherwise, the application of red Se⁰ resulted in a lower Se uptake; however, less fluctuation in total Se content between the four harvests was noticed compared to the ionic Se forms. The Se forms in stem and leaf of alfalfa extracted by water and subsequently by protease XIV enzyme were measured by strong anion exchange (SAX) HPLC-ICP-MS. The major Se forms in our samples were selenomethionine (SeMet) and Se (VI), while neither selenocysteine (SeCys) nor Se (IV) was detected. In water extract, however, Se (VI) was the major Se form, while SeMet was the predominant form in the enzyme extract. Yet, Se (VI) and SeMet contents declined within the harvests, except in stem of plants treated with 50 mg L⁻¹ red Se⁰. The highest stem or leaf SeMet yield %, in all harvests, corresponded to the treatment of 50 mg L⁻¹ red Se⁰. For instance, 63.6% (in stem) and 38.0% (in leaf) were calculated for SeMet yield % in the 4th harvest of plants treated with 50 mg L⁻¹ red Se⁰. Our results provide information about uptake and accumulation dynamics of different ionic Se forms in case of multiple-harvested alfalfa, which, besides being a good model plant, is an important target plant species in green biorefining.

Background and Aims: Wet processed alfalfa-based products can provide high-quality concentrated protein, also contain nutrients and phytochemicals. Agronomic fortification can increase selenium (Se) incorporation into green biomass. For this reason, the aim was to investigate how different forms of Se are incorporated into the green biomass of multiple-harvest alfalfa and how they affect the chemical quality of the processed product candidates. Methods: In this research, alfalfa was enriched with three forms of selenium at different concentrations [selenate (Se(VI)); selenite (Se(IV)); and red elemental Se (Se0)]. The fortified green biomass was fractionated into leaf protein concentrate (LPC), fiber and phytoserum, and changes in selenium content and speciation, protein and phytochemical composition were determined. Results: Total Se content in alfalfa fractions drastically reduced within the four consecutive harvests, particularly for Se(VI) and Se(IV) forms, and significantly varied according to the Se treatments. Selenomethionine (SeMet) was the predominant organic Se species in LPC (35.7–246.0 µg g −¹ DW) and fiber (9.0- 47.7 µg g −¹ DW) fractions. Se-fortification induced considerable changes in the crude protein of LPC, which varied between 29—45 (m/m)%. Phytochemical composition markedly varied among Se-treatments. Besides several flavonoids, aglycones and glycosyl derivatives, apigenin glucuronide, and apigenin were the most abundant with a concentration up to 170.85 µg g −¹ DW. Medicagenic acid concentration was below 1.86 µg g −¹ DW in LPC. Conclusions: Se-enhanced alfalfa green biomass could provide functional products, such as LPC, for human and animal consumption with enhanced nutritional value, including Se, protein, and phytochemical contents.

Aims  To assess the distribution of the insertion/deletion (I/D) polymorphism of the angiotensin‐converting enzyme (ACE) gene in children and adolescents with Type 1 diabetes and to evaluate the association between ACE genotype and blood pressure (BP).Methods  ACE genotypes were assessed in 124 normoalbuminuric, clinically normotensive Type 1 diabetic children and adolescents and 120 non‐diabetic controls using polymerase chain reaction. Twenty‐four‐hour ambulatory BP monitoring was undertaken in all patients.Results  ACE genotypes distributed in patients as follows: 34 (27%) DD, 57 (46%) ID, 33 (27%) II. The distribution was similar in the control group: DD in 28% (33), ID in 45% (54), and II in 27% (33). Patients with DD genotype had higher mean 24‐h diastolic BP (73.8 ± 6.2 vs. 70.2 ± 5.0 and 69.7 ± 6.3 mmHg; P = 0.005) and lower diurnal variation in BP (11.8 ± 4.6 vs. 14.2 ± 4.2 and 14.8 ± 4.3%; P = 0.011) compared with ID and II groups. Four patients in the DD group proved to be non‐dipper compared with one in the ID and none in the II group (P = 0.026). Twenty‐four‐hour diastolic blood pressure was independently predictive for AER as dependent variable in the DD genotype patient group (r² = 0.12, P = 0.03).Conclusions  Children and adolescents with Type 1 diabetes do not differ from the non‐diabetic population regarding the I/D polymorphism of the ACE gene. ACE gene polymorphism is associated with BP abnormalities in normotensive and normoalbuminuric children and adolescents with Type 1 diabetes.

One of the aims of this study was to improve the sample throughput of a microwave-assisted closed vessel digestion system by using small quartz vials in polytetrafluoroethylene (PTFE) vessels for the sample preparation of raw propolis samples in small amounts. The digested samples were measured by inductively coupled plasma optical emission spectrometry (ICP-OES) and inductively coupled plasma mass spectrometry (ICP-MS) analyzing 36 elements. Limit of detection was low enough to measure all the elements, with the exception of La, in all raw propolis samples. There were no cross-contamination between the quartz vials, therefore independent samples can be prepared in the same PTFE vessel. Accuracy of the method was checked by spike recoveries and by analyzing BCR 189 wholemeal flour and two plant samples from a collaborative study. The means of RSDs were 5.3%–14.4% in the case of measured elements. The sample throughput was increased by three times using quartz vials in PFTE vessels besides matching with the requirements of green chemistry. Another goal was the characterization of the element content and thereby geographical identification of Hungarian raw propolis. In total, 252 samples were analyzed and their statistical characteristics were described. We cannot find globally such results of propolis element content, which is representing one country and with such a number of elements and samples. All the elements have positive skew and positive kurtosis. Concentration range is above two orders of magnitude in the case of Ba, Zn, V, Cr, Ni, Cd and Eu elements. The decimal logarithm of element concentrations was used for geographical identification of raw propolis samples originating from seven regions of Hungary by linear discriminant analysis (LDA). Grouping of the samples of the Northern Great Plain was the most effective with 96.3% and 77.8% based on the original method and the cross validation, respectively. The same indicators for all the groups are 76.6% and 61.5%.

Background and aims: Selenium (Se)-biofortified foods are able to compensate for Se deficient diets, but the proportion of different Se species varies among plant species, and the bioavailability of Se species also varies. This study aims to examine the Se species composition of four vegetables. Methods: The effect of Se-enriched irrigation water on the concentrations of Se species in the edible parts of cabbage, carrots, tomatoes and green peas grown on different soil types was investigated and quantified using HPLC-ICP-MS with either standard addition or isotope dilution. Results: Cabbage leaf and carrot root contained a high proportion (88 and 92%, respectively) of selenate (SeVI), while in tomato fruit and green pea seed selenomethionine (SeMet) was found in the highest proportion (33% and 48%, respectively), but elemental Se was also detected. The elemental Se concentration of peas was significantly higher on sandy soil (2.29 mg kg⁻¹) than on sandy silt (1.38 mg kg⁻¹) or silt soil (1.34 mg kg⁻¹). In 100 g of fresh edible parts, the largest amount of Se species was found in cabbage leaves, but organic Se in the form of SeMet was the most abundant in green peas, being significantly higher than in carrots or tomatoes. Tomatoes contained a nearly three-fold ratio of organic Se to inorganic Se, the latter being mostly comprised of the elemental form, which is unusually high in vegetables. The highest rate of unknown Se species exceeded 66% in the case of peas, while the highest identification rate (> 95%) of Se could be achieved in tomato. Conclusion: Tomatoes converted the absorbed Se into organic Se in the highest ratio, while green peas had the highest amount of organic Se in fresh biomass, so biotransformation was the most effective for these vegetables without any significant biomass reduction. These vegetables may be adapted to soils with a higher Se content.