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Increasing the concentration of an element in edible produce (i.e., biofortification) can mitigate the element deficiency in humans. Sprouts are small but popular part of healthy diets providing vitamins and essential elements throughout the year. Element composition of sprouts can easily be amended, e.g., by soaking the grains in element-rich solution before germination (grain-priming). In addition, pre-treatment of grains to improve element translocation from the solution into the grain may further enhance the element concentration in the sprout. Cold plasma technique could provide such solution, as it increases wettability and water uptake of grains. Grains of common buckwheat (Fogopyrum esculentum Moench) were pre-treated/ untreated with cold plasma and soaked in ZnCl2 solution/pure water. Germination tests, α-amylase activity, grain hydrophilic properties and water uptake were assessed. Element composition of grain tissues and of sprouts was assessed by micro-particle-induced-X-ray emission and X-ray fluorescence spectroscopy, respectively. Grain-priming increased Zn concentration in shoots of common buckwheat sprouts more than five-times, namely from 79 to 423 mg Zn kg−1 dry weight. Cold plasma treatment increased grain wettability and water uptake into the grain. However, cold plasma pre-treatment followed by grain-priming with ZnCl2 did not increase Zn concentration in different grain tissues or in the sprouts more than the priming alone, but rather decreased the Zn concentration in sprout shoots (average ± standard error: 216 ± 6.13 and 174 ± 7.57 mg Zn kg−1 dry weight, respectively). When the fresh weight portion of whole sprouts (i.e., of roots and shoots) was considered, comparable average requirements of Zn, namely 24.5 % and 35 % for adult men and women would be satisfied by consuming cold plasma pre-treated and not pre-treated grains. Potential advantages of cold plasma pre-treatment need to be tested further, mainly to optimize the duration of soaking required to produce Zn-enriched sprouts.

Type 1 diabetes is a challenging disease, characterized by dynamic changes in the insulin need during life periods, seasons of the year, but also by everyday situations. In particular, changes in insulin need are evident before, during and after exercise and having meals. In the midst of different life demands, it can be very burdensome to achieve tight glycemic control to prevent late diabetes complications, and at the same time, to avoid hypoglycemia. Consequently, many individuals with type 1 diabetes are faced with diabetes distress, decreasing profoundly their quality of life. Today, the nationwide Finnish Diabetic Nephropathy (FinnDiane) Study, launched in 1997, has gathered data from more than 8,000 well-characterized individuals with type 1 diabetes, recruited from 93 centers all over Finland and has established its position as the world’s leading project on studying complications in individuals with type 1 diabetes. Studying risk factors and mechanisms of diabetes complications is inconceivable without trying to understand the effects of exercise and nutrition on glycemic control and the development of diabetes complications. Therefore, in this paper we provide findings regarding food and exercise, accumulated during the 25 years of studying lives of Finnish people with type 1 diabetes.

The despeckling of synthetic aperture radar images using two different convolutional neural network architectures is presented in this paper. The first method presents a novel Siamese convolutional neural network with a dilated convolutional network in each branch. Recently, attention mechanisms have been introduced to convolutional networks to better model and recognize features. Therefore, we propose a novel design for a convolutional neural network using an attention mechanism for an encoder–decoder-type network. The framework consists of a multiscale spatial attention network to improve the modeling of semantic information at different spatial levels and an additional attention mechanism to optimize feature propagation. Both proposed methods are different in design but they provide comparable despeckling results in subjective and objective measurements in terms of correlated speckle noise. The experimental results are evaluated on both synthetically generated speckled images and real SAR images. The methods proposed in this paper are able to despeckle SAR images and preserve SAR features.

This paper presents the design of a high-voltage pulse-based radar and a supervised data processing method for soil moisture estimation. The goal of this research was to design a pulse-based radar to detect changes in soil moisture using a cross-hole approach. The pulse-based radar with three transmitting antennas was placed into a 12 m deep hole, and a receiver with three receive antennas was placed into a different hole separated by 100 m from the transmitter. The pulse generator was based on a Marx generator with an LC filter, and for the receiver, the high-frequency data acquisition card was used, which can acquire signals using 3 Gigabytes per second. Used borehole antennas were designed to operate in the wide frequency band to ensure signal propagation through the soil. A deep regression convolutional network is proposed in this paper to estimate volumetric soil moisture using time-sampled signals. A regression convolutional network is extended to three dimensions to model changes in wave propagation between the transmitted and received signals. The training dataset was acquired during the period of 73 days of acquisition between two boreholes separated by 100 m. The soil moisture measurements were acquired at three points 25 m apart to provide ground truth data. Additionally, water was poured into several specially prepared boreholes between transmitter and receiver antennas to acquire additional dataset for training, validation, and testing of convolutional neural networks. Experimental results showed that the proposed system is able to detect changes in the volumetric soil moisture using Tx and Rx antennas.

With shorter wavelengths than microwaves and greater penetration depth than infrared light, waves in the terahertz spectrum offer unique material testing opportunities. Terahertz technology offers non-invasive and non-destructive testing in the form of spectroscopy and imaging. The most used systems for terahertz imaging are time-domain spectroscopy systems. However, frequency domain spectroscopy systems could offer excellent frequency resolution and be more suitable for biomedical applications. Terahertz imaging based on frequency domain spectroscopy systems is slow, and suffers from frequency tuning errors. A novel one-dimensional imaging principle is presented in this paper. In addition, frequency range optimization based on convolutional neural networks and occlusion sensitivity is utilized for frequency range optimization. Frequency range optimization is used to determine the optimal frequency range for data acquisition. The optimal frequency range or bandwidth should be wide enough for effective phase detection, and should be at the intersection of several spectral footprints in the observed medium. The intersection of spectral footprints is estimated using the proposed frequency range optimization algorithm based on a convolutional neural network and occlusion sensitivity algorithm. The proposed algorithm selects the most sensitive frequency band of THz spectrum automatically, and enables very fast acquisitions for object inspection and classification.

Reactive postprandial hypoglycemia (RPH) is an understudied condition that lacks clinical definition, knowledge of future health implications, and an understanding of precise underlying mechanisms. Therefore, our study aimed to assess the glycemic response after glucose ingestion in individuals several years after the initial evaluation of RPH and to compare glucose regulation in individuals with RPH vs. healthy volunteers. We assessed the inter- and intra-individual differences in glucose, insulin, and C-peptide concentrations during 5-h oral glucose tolerance tests (OGTTs); the surrogate markers of insulin resistance (HOMA-IR and Matsuda index); and beta-cell function (distribution index and insulinogenic index). The study included 29 subjects with RPH (all females, aged 39 (28, 46) years) and 11 sex-, age-, and body mass index (BMI)-matched controls. No biochemical deterioration of beta-cell secretory capacity and no progression to dysglycemia after 6.4 ± 4.2 years of follow-up were detected. RPH subjects were not insulin resistant, and their insulin sensitivity did not deteriorate. RPH subjects exhibited no differences in concentrations or in the shape of the glucose-insulin curves during the 5-h OGTTs compared to age- and BMI-matched controls. No increased incident type 2 diabetes risk indices were evident in individuals with RPH. This dictates the need for further research to investigate the magnitude of future diabetes risk in individuals experiencing RPH.

This article addresses Croatian scholars’ information behavior and how they use technology to acquire information in three areas of their work: teaching, research, and administrative activities. Our study aims to find which communication channels scholars utilize to find and share knowledge. Are they using communication channels targeting a broader audience, i.e., formal–explicit communication, or those targeting a narrower one, i.e., informal–implicit communication? The questionnaire used included four questions regarding scholar activities, with nine possible communication channels, scored on a seven-point Likert scale. Considering many channels for each area of activity, a reduction was made through Principal Component Analysis (PCA), to determine latent components in various channels. In finding information for teaching activities, the main communication channel is informal and implicit, while for research and administrative activities, it is formal and explicit. PCA shows a distinction between social and technical domains of science in terms of how scholars collect material for administrative tasks. A further communication channel is reduced to two factors for all questions, where the first factor has formal–explicit and the second has informal–implicit characteristics. This work is part of a larger study aimed at determining the mechanisms of information diffusion within academic institutions, utilizing the Information space model.

This study was designed to evaluate the nano–bio interactions between endogenous biothiols (cysteine and glutathione) with biomedically relevant, metallic nanoparticles (silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs)), in order to assess the biocompatibility and fate of nanoparticles in biological systems. A systematic and comprehensive analysis revealed that the preparation of AgNPs and AuNPs in the presence of biothiols leads to nanoparticles stabilized with oxidized forms of biothiols. Their safety was tested by evaluation of cell viability, reactive oxygen species (ROS) production, apoptosis induction and DNA damage in murine fibroblast cells (L929), while ecotoxicity was tested using the aquatic model organism Daphnia magna. The toxicity of these nanoparticles was considerably lower compared to their ionic metal forms (i.e., Ag+ and Au3+). The comparison with data published on polymer-coated nanoparticles evidenced that surface modification with biothiols made them safer for the biological environment. In vitro evaluation on human cells demonstrated that the toxicity of AgNPs and AuNPs prepared in the presence of cysteine was similar to the polymer-based nanoparticles with the same core material, while the use of glutathione for nanoparticle stabilization was considerably less toxic. These results represent a significant contribution to understanding the role of biothiols on the fate and behavior of metal-based nanomaterials.

Abstract Background Both obesity and gestational diabetes (GDM) are associated with adverse outcomes. Diet during pregnancy impacts weight gain and fetal growth. Therefore, we aimed to explore non-pharmacological treatment success depending on pre-pregnancy body weight and its association with large for gestational age (LGA) infants in women with GDM. Methods In our observational study we investigated 57 singleton pregnant women with GDM. All women received standard treatment, including healthy diet education and regular medical checkups. Data were collected through blood analysis, medical records and questionnaires assessing diet before conception and during pregnancy. Differences in dietary patterns were compared in normal weight and overweight/obese group using Mann–Whitney U, Wilcoxon Signed Rank Test or Kruskal–Wallis test, as appropriate. Logistic regression was used for prediction of LGA. p-value less than 0.05 was used for statistical significance. Results Preconceptionally, the Mann–Whitney U test showed that the normal-weight group (n = 41) more frequently consumed fruits (U = 116.5, p

The study focused on the relationship of emotional and cognitive components of empathy with emotional intelligence abilities (labelling, understanding, and regulating emotions). Since both emotional intelligence and empathy have important role in a person’s socio-emotional health and well-being, we need to better understand the relations between these two related constructs. Additionally, having samples from two different countries enabled us comparison on the national level. Two samples of adults participated in the study, namely from Croatia (N = 138) and Portugal (N = 138). We found significant moderate correlations between empathy and emotional intelligence measures in both samples. Four dimensions from the Interpersonal Responsivity Index (IRI; empathic concerns, fantasy, personal distress and perspective-taking) had significant moderate correlations with each other, with similar patterns of correlations in both samples. The strongest correlations were found for the TEQ (affective component of empathy) and perspective-taking (cognitive component of empathy) and different emotional intelligence components. Participants in the Portuguese sample had higher scores on the emotional component of empathy (empathic care and the TEQ), while cognitive component (perspective-taking) was more expressed in the Croatian sample. Participants in the Portuguese sample also had higher scores on all three components of emotional intelligence than participants in the Croatian sample. Regardless of the nationality, women had higher scores on the measures of empathic care, fantasy, perspective-taking and the TEQ, while men scored higher on the measure of personal distress. Regarding emotional intelligence scales, women had higher results on the Perceive and Understand emotions scale.

The article clarifies the importance of maintenance in the exploitation of a particular medical device from the aspect of maintaining its availability, reliability and overall functionality. Particularly emphasized is the role of the preventive method or procedures that have a preventive character before the problems arise in the work. Through an analysis of the most common malfunctions and causes that occur during the exploitation, a basis for objective defining and proposing principles has been created – preventive maintenance with special emphasis on preventive maintenance according to an established condition. The article also shows a numerical way of calculating the periodicity of conducting preventive examinations on a particular device.

Shoot zinc (Zn) concentration in Brassica oleracea is affected by soil Zn and phosphorus (P) supply. Most problematic is the negative impact of P fertilizers on Zn concentrations in crops, which makes balancing yield and mineral quality challenging. To evaluate early molecular mechanisms involved in the accumulation of large shoot Zn concentrations regardless of the P supply, two B. oleracea accessions differing in root architecture and root exudates were grown hydroponically for two weeks with different combinations of P and Zn supply. Ionome profiling and deep RNA sequencing of roots revealed interactions of P and Zn in planta, without apparent phenotypic effects. In addition, increasing P supply did not reduce tissue Zn concentration. Substantial changes in gene expression in response to different P and/or Zn supplies in roots of both accessions ensured nutritionally sufficient P and Zn uptake. Numerous genes were differentially expressed after changing Zn or P supply and most of them were unique to only one accession, highlighting their different strategies in achieving nutrient sufficiency. Thus, different gene networks responded to the changing P and Zn supply in the two accessions. Additionally, enrichment analysis of gene ontology classes revealed that genes involved in lipid metabolism, response to starvation, and anion transport mechanisms were most responsive to differences in P and Zn supply in both accessions. The results agreed with previously studies demonstrating alterations in P and Zn transport and phospholipid metabolism in response to reduced P and Zn supply. It is anticipated that improved knowledge of genes responsive to P or Zn supply will help illuminate the roles in uptake and accumulation of P and Zn and might identify candidate genes for breeding high-yield-high-Zn brassicas.

This study investigates the effect of combined sulfur (S) and nitrogen (N) foliar fertilization on leaf S and N concentration, as well as on the growth of olive fruit and on the quantity and quality of olive oil, obtained from two olive cultivars ‘Istarska bjelica’ and ‘Leccino’ in two consecutive years. S and N are some of the most important nutrients, and both play a crucial role in plant oil production. The here-reported fertilization program significantly increased S concentration in leaves without affecting N concentration, which led to an increase in fruit yield and improvement of all fruit morphological parameters. The best oil yield per tree was obtained under the treatment with the highest S/N dose. Oil quality was not affected by S and N supply, and this allowed us to classify all our oil samples as extra virgin (EVOO). Regarding the content of total phenols (TPC) and composition of fatty acid methyl esters (FAME), they remained unaltered under the applied treatments. All investigated fruit morphological parameters, as well as fruit and oil yield, were highly cultivar-dependent. ‘Istarska bjelica’ was characterized as a cultivar with higher fruit mass and pulp percentage, while its stone parameters were lower than those of ‘Leccino’. Consequently, the extraction oil yield obtained from ‘Istarska bjelica’ fruits was much higher. Moreover, environmental conditions had a great impact on fruit and oil quantity. The here-obtained results led us to the conclusion that supply of S and N can enhance oil production without affecting its nutritional quality, a finding that could generate large long-term effects on economic growth in the olive oil sector.

During the period 2004–2015, the upper soil layer (0–20 cm) was sampled in eight agricultural production areas in Slovenia. Overall, 53, 249, 169, 139, and 413 samples were respectively taken from vegetable fields, arable cropland, hop fields, orchards, and vineyards. Copper was measured in the aqua-regia extracts of these samples using flame atomic absorption spectrometry. The minimum, maximum, average, and median concentrations were calculated for five land uses and for all eight production areas in Slovenia. Sample portions below and/or above the limit, warning, and critical concentrations of 60, 100, and 300 mg/kg, respectively, were calculated as well. The results were compared for different land uses and production areas and showed that the copper concentrations in the vegetable, arable crop, and hop fields and orchards did not exceed the critical concentration. In addition, the copper concentrations in the vineyards exceeded the critical concentration but the maximum copper concentration (508 mg/kg) was still lower than the concentrations reported from other vine-growing Mediterranean countries. Also, the copper concentrations in hop fields exhibited an increasing trend in the Celje (and Maribor) area(s).

Magnetic resonance imaging (MRI) of superparamagnetic iron oxide-labeled cells can be used as a non-invasive technique to track stem cells after transplantation. The aim of this study was to (1) evaluate labeling efficiency of D-mannose-coated maghemite nanoparticles (D-mannose(γ-Fe 2 O 3 )) in neural stem cells (NSCs) in comparison to the uncoated nanoparticles, (2) assess nanoparticle utilization as MRI contrast agent to visualize NSCs transplanted into the mouse brain, and (3) test nanoparticle biocompatibility. D-mannose(γ-Fe 2 O 3 ) labeled the NSCs better than the uncoated nanoparticles. The labeled cells were visualized by ex vivo MRI and their localization subsequently confirmed on histological sections. Although the progenitor properties and differentiation of the NSCs were not affected by labeling, subtle effects on stem cells could be detected depending on dose increase, including changes in cell proliferation, viability, and neurosphere diameter. D-mannose coating of maghemite nanoparticles improved NSC labeling and allowed for NSC tracking by ex vivo MRI in the mouse brain, but further analysis of the eventual side effects might be necessary before translation to the clinic.

Background: Cell tracking is a powerful tool to understand cellular migration, dynamics, homing and function of stem cell transplants. Nanoparticles represent possible stem cell tracers, but they differ in cellular uptake and side effects. Their properties can be modified by coating with different biocompatible polymers. To test if a coating polymer, poly(L-lysine), can improve the biocompatibility of nanoparticles applied to neural stem cells, poly(L-lysine)-coated maghemite nanoparticles were prepared and characterized. We evaluated their cellular uptake, the mechanism of internalization, cytotoxicity, viability and proliferation of neural stem cells, and compared them to the commercially available dextran-coated nanomag®-D-spio nanoparticles.Results: Light microscopy of Prussian blue staining revealed a concentration-dependent intracellular uptake of iron oxide in neural stem cells. The methyl thiazolyl tetrazolium assay and the calcein acetoxymethyl ester/propidium iodide assay demonstrated that poly(L-lysine)-coated maghemite nanoparticles scored better than nanomag®-D-spio in cell labeling efficiency, viability and proliferation of neural stem cells. Cytochalasine D blocked the cellular uptake of nanoparticles indicating an actin-dependent process, such as macropinocytosis, to be the internalization mechanism for both nanoparticle types. Finally, immunocytochemistry analysis of neural stem cells after treatment with poly(L-lysine)-coated maghemite and nanomag®-D-spio nanoparticles showed that they preserve their identity as neural stem cells and their potential to differentiate into all three major neural cell types (neurons, astrocytes and oligodendrocytes).Conclusion: Improved biocompatibility and efficient cell labeling makes poly(L-lysine)-coated maghemite nanoparticles appropriate candidates for future neural stem cell in vivo tracking studies.

This paper will perform a multi-criteria analysis of the insurance sector in the Republic of Croatia. The analysis is performed based on five indicators for the eight largest insurance companies in Croatia. The multicriteria business performance is calculated with use of a score, the weighted sum of relative values of indicators. The score is obtained by calculating weights as a solution of a goal programming problem. The goal of a specific insurance company is to increase the value of an indicator in comparison to it’s value a year before. Hence, some insurance companies have more goals that they wish to obtain. The results reveal that the Croatian insurance sector has given the highest priority to return on investment and return on equity for 2011.

This paper studies the problem of interdependence between central bank and commercial bank goals. The basic central bank task is to achieve and to maintain price stability. Croatian external debt has been increasing for years and so the activities of the Croatian National Bank are designed to correct this situation. In order to stop the further increase of the external debt, the Croatian National Bank uses several monetary policy instruments, among which is the marginal reserve requirement. On the other hand, the goal of commercial banks is to maximise profits. Banks take loans from abroad at a lower interest rate and invest this money in Croatia at a higher interest rate, thus fulfilling their goal. In order to obtain the desired effects of the marginal reserve requirement, its optimal percentage value should be determined. This problem is modelled as a bi-level mixed 0-1 programming problem. The objective of the leader (Croatian National Bank) is to minimize the increase in household loans by setting different percentages of the reserve requirements for loans extended to households and for those granted to enterprises. The objective of the followers (banks) is to maximize profits. In order to solve this NP-hard problem a heuristic is proposed. In order to verify the model, the paper ends with simulations and the presentation of computational results.

To add new evidence that complement activation by the mannan (lectin) pathway is involved in the pathogenesis of nephropathy in patients with type 2 diabetes, we collected blood and urine samples from 20 patients with type 2 diabetes and nephropathy and 20 apparently healthy individuals from the general population. We performed tests for complement activation analysis (CH50, APH50, C3, C4, C3d/dg, C4d, SC5b-9, MBL and genotyping of Mbl2 gene) in blood and (C3d/dg, C4d, MBL, and SC5b-9) in urine. We found significant signs of complement activation in patients with type 2 diabetes and nephropathy in blood and in urine. We also observed significant genetic differences between healthy individuals and patients with type 2 diabetes and nephropathy concerning the Mbl2 gene. Healthy individuals were apparently more heterogeneous (45 % A0 type) in presentation of the structural Mbl2 genotype, giving intermediate and low levels of MBL protein, compared to patients with type 2 diabetes and nephropathy, who had a very homogenous Mbl2 genome (75 % AA type) giving a high level of MBL production. From our data we can conclude that patients with type 2 diabetes and nephropathy presents more frequently with Mbl2 gene polymorphism giving high production of the MBL protein, what is very likely connected with acquired renal injury.

The impact of simultaneous Cd and Zn amendments in the substrate on the accumulation of Cd and Zn were studied in a recently discovered Cd/ Zn hyperaccumulating Thlaspi praecox (Brassicaceae) and compared to a model hyperaccumulating plant species T. caerulescens. The plants were grown in pots with added Cd or Zn or both for three months in a greenhouse. The addition of Zn in the substrate increased Cd extractability in the substrate significantly without a significant pH change and this increase resulted in increased concentration and content of Cd in the shoots of both species indicating that species have similar abilities to extract Cd from the substrate. In the combined treatment (Cd and Zn) an increase in shoot biomass accompanied with a decrease in Zn concentration in roots and shoots of both species was observed, while no changes in total accumulated Zn in shoots were seen. These results suggest different uptake and translocation systems for Cd and Zn in T. praecox, positioning this plant species in the superior Cd hyperaccumulating league of T. caerulescens Ganges ecotype.

Leta 1983 smo zasnovali večletni gnojilni poskus po načelu naključnih blokov v štirih ponovitvah. V letu 1994 smo zbrali vzorce krme na negnojeni različici (0), različici, gnojeni s fosforjem in kalijem (PK), ter različici, gnojeni s fosforjem, kalijem in dušikom (NPK). Vzorce prve, druge in tretje košnje smo ločili na trave, metuljnice in zeli ter ločeno določali vsebnosti Ca, P, Mg, K, Na, Zn, Mn, Fe, in Cu. Glede na delež posamezne botanične skupine smo izračunali tudi vsebnosti elementov v krmi. Gnojenje je značilno vplivalo na vsebnost večine elementov v travah, metuljnicah in zeleh vseh treh košenj. Gnojenje s PK je v krmi zmanjšalo vsebnost Mg in Mn ter povečalo vsebnost P in K. Pri gnojenju z NPK v primerjavi z negnojeno različico (0) se je v krmi zmanjšala vsebnost Fe, povečala pa vsebnost P, K in Zn. Gnojenje z dušikom (NPK vs. PK) je v krmi značilno povečalo vsebnost P, Mg, Zn in Mn ter zmanjšalo vsebnost Ca. Zaporedna košnja je značilno vplivala na vsebnost vseh analiziranih elementov v krmi razen na vsebnost Cu. Pri krmi druge in tretje košnje je bila vsebnost Ca, Mg in Mn večja kot pri krmi prve košnje. Največje vsebnosti P in Zn smo ugotovili v krmi tretje košnje. Metuljnice in zeli vsebujejo v primerjavi s travami dva- do trikrat več Ca, Mg, Zn in Cu. Zaradi tega se s spreminjanjem botanične sestave travne ruše precej spreminja tudi vsebnost teh elementov v krmi. Glede na potrebe krav v laktaciji smo ugotovili izrazito pomanjkanje Na, Zn in Cu, pomanjkanje P pa le v krmi z negnojenih parcel.

Ruminal starch digestibility of dried maize grain and whole plant maize silage was determined by the in sacco method. It was found out that starch of dent type maize hybrid is more extensively digested in the rumen than starch of flint type hybrid. Digestibilities of dried maize grain from dent and flint type hybrids were 683 and 554 g kg-1 respectively. Ruminal digestibilities of corresponding silages were 915 and 743 g kg-1. Ruminal digestibility of starch in whole plant maize silage was on average about 200 g kg-1 higher than in dried grain samples. Ruminal starch digestibility of maize silages can not be estimated directly on the basis of measurements done on dried grain samples. Some practical implications of differences among hybrids and between dried and ensiled maize are discussed.

Hyperaccumulators within the Noccaea genus possess many promising genetic and metabolic adaptations that could be potentially exploited to support phytoremediation efforts and/or crop improvement and biofortification. Although hyperaccumulation is very common in this genus, individual species display specific traits as they can accumulate different elements (e.g., zinc, cadmium, and/or nickel). Moreover, there appears to be some populational variability with natural selection increasing the metal tolerance in metallicolous populations. Therefore, employing robust methods, such as integrated analysis of the transcriptome and metabolome, is crucial for uncovering pivotal candidate genes and pathways orchestrating the response to metal stress in Noccaea hyperaccumulators. Our study highlights several species-specific traits linked to the detoxification of metals and metal-induced oxidative stress in hyperaccumulating N. praecox when compared to a closely related model species, N. caerulescens , when grown in the field. Transcriptome analysis revealed distinct differences between the three studied natural Noccaea populations. Notably, we observed several pathways frequently connected to metal stress, i.e., glutathione metabolism, phenylpropanoid biosynthesis, and flavonoid biosynthesis, which were enriched. These differences were observed despite the relative evolutionary closeness of studied species, which emphasizes the importance of further expanding our knowledge on hyperaccumulators if we want to exploit their mechanisms for phytoremediation efforts or food quality improvements.

The quality of molecular imaging by means of MeV primary ion-induced secondary ion mass spectrometry by coating with gold was evaluated on different reference organic molecules and plant samples. The enhancement of the secondary ion yield was evident for the majority of the studied analytes, reaching the highest values at gold thicknesses between 0.5 and 2 nm, and increased the intensity up to 5-fold for reference samples and >2-fold for specific peaks within the plant sample. Improved propagation of the electric field due to the target potential on otherwise electrically insulating plant samples was also evident through improved image resolution and by reducing the background in mass spectra. However, detection of several molecules was significantly decreased at even at 1 nm thick gold layer. The results indicated that an optimized sequence of analysis is required to reliably interpret results.

Polygalacturonases (PGs) fine-tune pectins to modulate cell wall chemistry and mechanics, impacting plant development. The large number of PGs encoded in plant genomes leads to questions on the diversity and specificity of distinct isozymes. Herein, we report the crystal structures of 2 Arabidopsis thaliana PGs, POLYGALACTURONASE LATERAL ROOT (PGLR), and ARABIDOPSIS DEHISCENCE ZONE POLYGALACTURONASE2 (ADPG2), which are coexpressed during root development. We first determined the amino acid variations and steric clashes that explain the absence of inhibition of the plant PGs by endogenous PG-inhibiting proteins (PGIPs). Although their beta helix folds are highly similar, PGLR and ADPG2 subsites in the substrate binding groove are occupied by divergent amino acids. By combining molecular dynamic simulations, analysis of enzyme kinetics, and hydrolysis products, we showed that these structural differences translated into distinct enzyme-substrate dynamics and enzyme processivities: ADPG2 showed greater substrate fluctuations with hydrolysis products, oligogalacturonides (OGs), with a degree of polymerization (DP) of ≤4, while the DP of OGs generated by PGLR was between 5 and 9. Using the Arabidopsis root as a developmental model, exogenous application of purified enzymes showed that the highly processive ADPG2 had major effects on both root cell elongation and cell adhesion. This work highlights the importance of PG processivity on pectin degradation regulating plant development., Competing Interests: Conflict of interest statement. None declared., (© American Society of Plant Biologists 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Increasing the concentration of an element in edible produce (i.e., biofortification) can mitigate the element deficiency in humans. Sprouts are small but popular part of healthy diets providing vitamins and essential elements throughout the year. Element composition of sprouts can easily be amended, e.g., by soaking the grains in element-rich solution before germination (grain-priming). In addition, pre-treatment of grains to improve element translocation from the solution into the grain may further enhance the element concentration in the sprout. Cold plasma technique could provide such solution, as it increases wettability and water uptake of grains. Grains of common buckwheat ( Fogopyrum esculentum Moench) were pre-treated/ untreated with cold plasma and soaked in ZnCl 2 solution/pure water. Germination tests, α-amylase activity, grain hydrophilic properties and water uptake were assessed. Element composition of grain tissues and of sprouts was assessed by micro-particle-induced-X-ray emission and X-ray fluorescence spectroscopy, respectively. Grain-priming increased Zn concentration in shoots of common buckwheat sprouts more than five-times, namely from 79 to 423 mg Zn kg -1 dry weight. Cold plasma treatment increased grain wettability and water uptake into the grain. However, cold plasma pre-treatment followed by grain-priming with ZnCl 2 did not increase Zn concentration in different grain tissues or in the sprouts more than the priming alone, but rather decreased the Zn concentration in sprout shoots (average ± standard error: 216 ± 6.13 and 174 ± 7.57 mg Zn kg -1 dry weight, respectively). When the fresh weight portion of whole sprouts (i.e., of roots and shoots) was considered, comparable average requirements of Zn, namely 24.5 % and 35 % for adult men and women would be satisfied by consuming cold plasma pre-treated and not pre-treated grains. Potential advantages of cold plasma pre-treatment need to be tested further, mainly to optimize the duration of soaking required to produce Zn-enriched sprouts., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Starič, Remic, Vogel-Mikuš, Junkar, Vavpetič, Kelemen and Pongrac.)

Olive tree (Olea europaea L.) leaves have recently been recognised as a valuable source in cosmetic and pharmaceutical industry as well as in preparation of health-supporting beverages. Little is known about the element composition of olive leaves and almost nothing about tissue-specific allocation of elements. Element composition and tissue-specific distribution were determined in leaves of two olive cultivars, Leccino and Istarska bjelica using micro-particle induced X-ray emission (micro-PIXE). In leaves of the Istarska bjelica cultivar larger bulk concentrations of potassium, sodium, molybdenum and boron, but smaller concentrations of calcium and magnesium were found than in leaves of the Leccino cultivar. Tissue-specific investigation revealed that larger concentration of calcium in epidermis and in leaf blade tissues (secondary veins, palisade and spongy mesophyll) contributed to the larger leaf bulk calcium concentration in the Leccino cultivar. For magnesium, all leaf tissues, except the bundle sheath cells and consequently the main vascular bundle, contributed to the larger bulk concentration in the Leccino cultivar. Potassium was not predominant in any of the leaf tissues examined, while sodium and molybdenum were below the limit of detection, and boron not detectable by micro-PIXE. The results indicate that sinks for calcium and magnesium are stronger in specific leaf tissues of the Leccino than of the Istarska bjelica cultivar. The new understanding of tissue-specific allocation of elements in leaves of olive will serve as a basis for detailed studies into the effects of foliar and/or soil fertilisers in olive., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Arbuscular mycorrhizal fungi (AMF) exhibit great potential in heavy-metal immobilization in semi-aquatic habitats. Under high heavy-metal stress, however, the role of AMF is limited, and the detoxification mechanism of AMF in heavy metals' stabilization remains unclear. This study investigated the effects of AMF on a wetland plant (Iris pseudacorus) and chromium (Cr) immobilization at different water depths in semi-aquatic habitats with biochar addition. Results showed that AMF increased the physiological and photosynthetic functions in I. pseudacorus under Cr exposures. Besides, AMF alleviated the accumulation of reactive oxygen species and lipid peroxidation by enhancing the antioxidant enzyme activities. AMF and biochar significantly decreased Cr concentrations in outlet water and increased Cr accumulation in I. pseudacorus. Besides, biochar also vastly improved Cr accumulation in the substrate under the fluctuating water depth. AMF reduced Cr bioavailability in the substrate, with Cr (Ⅵ) concentrations and acid-soluble forms of Cr decreased by 0.3-64.5% and 19.0-40.8%, respectively. Micro-proton-induced X-ray emission was used to determine element localization and revealed that AMF improved the nutrients uptake by wetland plants and inhibited Cr translocation from roots to shoots. Overall, this study demonstrated that the interaction between AMF and biochar could significantly enhance the immobilization of high Cr concentrations in semi-aquatic habitats., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Silkworm rearing activities ceased in the 1970's in several European countries. Attempts on the re-establishment of ecological and sustainable sericulture in Slovenia and Hungary are ongoing. The aim of the study was to assess the usability of locally adapted mulberry genotypes for sericulture and to estimate connections between leaf compound and silkworm performance parameters. A controlled feeding experiment of silkworms was performed to test the influence of leaves from selected trees on the growth of larvae, the health and microbiological status of larvae (e.g., gut bacterial microbiome, Bombyx mori nucleopolyhedrovirus infection), weight of cocoons and raw silk parameters. The Slovenian and Hungarian mulberry genotypes had significantly higher total protein contents, and lower total phenolic contents and differed significantly in some individual phenolics compared to the reference sericultural and fruit varieties. Significant differences were found in the contents of the macro- and microelements, namely S, Mn, Fe, and Sr. Based on correlative statistics and multivariate analysis, a combined positive influence of proteins, specific phenolics, and microelements on larval growth and silk thread parameters was predicted. The results of the study indicate that selected local Slovenian and Hungarian mulberry varieties are suitable for high-quality silk cocoon and raw silk production.

Salinity is a growing global concern that affects the yield of crop species, including tomato ( Solanum lycopersicum ). Its wild relative Solanum chilense was reported to have halophyte properties. We compared salt resistance of both species during the reproductive phase, with a special focus on sodium localization in the flowers. Plants were exposed to NaCl from the seedling stage. Salinity decreased the number of inflorescences in both species but the number of flowers per inflorescence and sepal length only in S. lycopersicum . External salt supply decreased the stamen length in S. chilense , and it was associated with a decrease in pollen production and an increase in pollen viability. Although the fruit set was not affected by salinity, fruit weight and size decreased in S. lycopersicum . Concentrations and localization of Na, K, Mg, and Ca differed in reproductive structures of both species. Inflorescences and fruits of S. chilense accumulated more Na than S. lycopersicum . Sodium was mainly located in male floral organs of S. chilense but in non-reproductive floral organs in S. lycopersicum . The expression of Na transporter genes differed in flowers of both species. Overall, our results indicated that S. chilense was more salt-resistant than S. lycopersicum during the reproductive phase and that differences could be partly related to dissimilarities in element distribution and transport in flowers.

In plants, the response to stress, such as salinity, pathogen attack, drought, high concentration of metals, hyperthermia, and hypothermia, is usually accompanied by potassium ion (K + ) leakage from the cytosol to the cell wall, mediated by plasma membrane cation conductivity. Stress-induced electrolyte leakage co-occurs with accumulation of reactive oxygen species (ROS) and calcium ions (Ca 2+ ) and often results in programmed cell death (PCD). The development of X-ray and mass spectrometry (MS) based imaging techniques has enabled insight into the spatial tissue and cell-specific redistribution of major and trace elements during the stress response. In this chapter a workflow for sample preparation, imaging, and image analysis by X-ray and MS based techniques is presented., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Magnesium (Mg) and calcium (Ca) are essential mineral nutrients poorly supplied in many human food systems. In grazing livestock, Mg and Ca deficiencies are costly welfare issues. Here, we report a Brassica rapa loss-of-function schengen3 (sgn3) mutant, braA.sgn3.a-1, which accumulates twice as much Mg and a third more Ca in its leaves. We mapped braA.sgn3.a to a single recessive locus using a forward ionomic screen of chemically mutagenized lines with subsequent backcrossing and linked-read sequencing of second back-crossed, second filial generation (BC2F2) segregants. Confocal imaging revealed a disrupted root endodermal diffusion barrier, consistent with SGN3 encoding a receptor-like kinase required for normal formation of Casparian strips, as reported in thale cress (Arabidopsis thaliana). Analysis of the spatial distribution of elements showed elevated extracellular Mg concentrations in leaves of braA.sgn3.a-1, hypothesized to result from preferential export of excessive Mg from cells to ensure suitable cellular concentrations. This work confirms a conserved role of SGN3 in controlling nutrient homeostasis in B. rapa, and reveals mechanisms by which plants are able to deal with perturbed shoot element concentrations resulting from a "leaky" root endodermal barrier. Characterization of variation in leaf Mg and Ca accumulation across a mutagenized population of B. rapa shows promise for using such populations in breeding programs to increase edible concentrations of essential human and animal nutrients., (© The Author(s) 2021. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

Common buckwheat ( Fagopyrum esculentum Moench) and Tartary buckwheat ( Fagopyrum tataricum (L.) Gaertn.) are sources of many bioactive compounds, such as rutin, quercetin, emodin, fagopyrin and other (poly)phenolics. In damaged or milled grain under wet conditions, most of the rutin in common and Tartary buckwheat is degraded to quercetin by rutin-degrading enzymes (e.g., rutinosidase). From Tartary buckwheat varieties with low rutinosidase activity it is possible to prepare foods with high levels of rutin, with the preserved initial levels in the grain. The quercetin from rutin degradation in Tartary buckwheat grain is responsible in part for inhibition of α-glucosidase in the intestine, which helps to maintain normal glucose levels in the blood. Rutin and emodin have the potential for antiviral effects. Grain embryos are rich in rutin, so breeding buckwheat with the aim of producing larger embryos may be a promising strategy to increase the levels of rutin in common and Tartary buckwheat grain, and hence to improve its nutritional value.

Seed phytic acid reduces mineral bioavailability by chelating minerals. Consumption of common bean seeds with the low phytic acid 1 (lpa1) mutation improved iron status in human trials but caused adverse gastrointestinal effects, presumably due to increased stability of lectin phytohemagglutinin L (PHA-L) compared to the wild type (wt). A hard-to-cook (HTC) defect observed in lpa1 seeds intensified this problem. We quantified the HTC phenotype of lpa1 common beans with three genetic backgrounds. The HTC phenotype in the lpa1 black bean line correlated with the redistribution of calcium particularly in the cell walls, providing support for the "phytase-phytate-pectin" theory of the HTC mechanism. Furthermore, the excess of free cations in the lpa1 mutation in combination with different PHA alleles affected the stability of PHA-L lectin., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Lead (Pb) ranks among the most problematic environmental pollutants. Background contamination of soils is nearly ubiquitous, yet plant Pb accumulation is barely understood. In a survey covering 165 European populations of the metallophyte Arabidopsis halleri, several field samples had indicated Pb hyperaccumulation, offering a chance to dissect plant Pb accumulation. Accumulation of Pb was analysed in A. halleri individuals from contrasting habitats under controlled conditions to rule out aerial deposition as a source of apparent Pb accumulation. Several elemental imaging techniques were employed to study the spatial distribution and ligand environment of Pb. Regardless of genetic background, A. halleri individuals showed higher shoot Pb accumulation than A. thaliana. However, dose-response curves revealed indicator rather than hyperaccumulator behaviour. Xylem sap data and elemental imaging unequivocally demonstrated the in planta mobility of Pb. Highest Pb concentrations were found in epidermal and vascular tissues. Distribution of Pb was distinct from that of the hyperaccumulated metal zinc. Most Pb was bound by oxygen ligands in bidentate coordination. A. halleri accumulates Pb whenever soil conditions render Pb phytoavailable. Considerable Pb accumulation under such circumstances, even in leaves of A. thaliana, strongly suggests that Pb can enter food webs and may pose a food safety risk., (© 2019 The Authors New Phytologist © 2019 New Phytologist Trust.)

Shoot zinc (Zn) concentration in Brassica oleracea is affected by soil Zn and phosphorus (P) supply. Most problematic is the negative impact of P fertilizers on Zn concentrations in crops, which makes balancing yield and mineral quality challenging. To evaluate early molecular mechanisms involved in the accumulation of large shoot Zn concentrations regardless of the P supply, two B. oleracea accessions differing in root architecture and root exudates were grown hydroponically for two weeks with different combinations of P and Zn supply. Ionome profiling and deep RNA sequencing of roots revealed interactions of P and Zn in planta , without apparent phenotypic effects. In addition, increasing P supply did not reduce tissue Zn concentration. Substantial changes in gene expression in response to different P and/or Zn supplies in roots of both accessions ensured nutritionally sufficient P and Zn uptake. Numerous genes were differentially expressed after changing Zn or P supply and most of them were unique to only one accession, highlighting their different strategies in achieving nutrient sufficiency. Thus, different gene networks responded to the changing P and Zn supply in the two accessions. Additionally, enrichment analysis of gene ontology classes revealed that genes involved in lipid metabolism, response to starvation, and anion transport mechanisms were most responsive to differences in P and Zn supply in both accessions. The results agreed with previously studies demonstrating alterations in P and Zn transport and phospholipid metabolism in response to reduced P and Zn supply. It is anticipated that improved knowledge of genes responsive to P or Zn supply will help illuminate the roles in uptake and accumulation of P and Zn and might identify candidate genes for breeding high-yield-high-Zn brassicas., (Copyright © 2020 Pongrac, Fischer, Thompson, Wright and White.)

In wheat ( Triticum aestivum L.), the awns-the bristle-like structures extending from lemmas-are photosynthetically active. Compared to awned cultivars, awnletted cultivars produce more grains per unit area and per spike, resulting in significant reduction in grain size, but their mineral element composition remains unstudied. Nine awned and 11 awnletted cultivars were grown simultaneously in the field. With no difference in 1000-grain weight, a larger calcium and manganese-but smaller iron (Fe) concentrations-were found in whole grain of awned than in awnletted cultivars. Micro X-ray absorption near edge structure analysis of different tissues of frozen-hydrated grain cross-sections revealed that differences in total Fe concentration were not accompanied by differences in Fe speciation (64% of Fe existed as ferric and 36% as ferrous species) or Fe ligands (53% were phytate and 47% were non-phytate ligands). In contrast, there was a distinct tissue-specificity with pericarp containing the largest proportion (86%) of ferric species and nucellar projection (49%) the smallest. Phytate ligand was predominant in aleurone, scutellum and embryo (72%, 70%, and 56%, respectively), while nucellar projection and pericarp contained only non-phytate ligands. Assuming Fe bioavailability depends on Fe ligands, we conclude that Fe bioavailability from wheat grain is tissue specific.

During tea preparation mineral elements are extracted from the dried leaves of tea (Camellia sinensis (L.) Kuntze) plants into the solution. Micro-particle induced X-ray emission was employed to investigate the spatial distribution of magnesium (Mg), calcium (Ca) and manganese (Mn) in the young and old leaves of tea plants grown in the absence and presence of aluminium (Al) in the substrate. Results revealed that in tea leaves the largest concentrations of Mg occurred in the epidermis, of Ca in oxalate crystals and of Mn in epidermis and oxalate crystals; there was a leaf-age effect on tissue-specific concentrations of Mg, Ca and Mn with all tissues of old leaves containing larger concentrations of Mg, Ca and Mn than young leaves; supplementation of substrate with Al reduced concentrations of Mg, Ca and Mn in the old leaves, and a link between the distribution of Mg, Ca and Mn in the tea leaves with the extraction efficiencies of these elements into the tea was possible. We conclude that old leaves of tea plants cultivated under conditions of low Al availability will have the largest concentrations of Mg, Ca and Mn and may represent most acceptable ingredient for the preparation of tea., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The fragmentary information on phosphorus (P) × zinc (Zn) interactions in plants warrants further study, particularly in plants known for their high P and Zn requirements, such as cotton (Gossypium hirsutum L.). The objective of this study was to investigate the effect of P × Zn interactions in a modern cultivar of cotton grown hydroponically. Biomass, mineral nutrition and photosynthetic parameters were monitored in plants receiving contrasting combinations of P and Zn supply. Root biomass, length and surface area were similar in plants with low P and/or low Zn supply to those in plants grown with high P and high Zn supply, reflecting an increased root/shoot biomass quotient when plants lack sufficient P or Zn for growth. Increasing P supply and reducing Zn supply increased shoot P concentrations, whilst shoot Zn concentrations were influenced largely by Zn supply. A balanced P × Zn supply (4 mM P × 4 μM Zn) enabled greatest biomass accumulation, while an imbalanced supply of these nutrients led to Zn deficiency, P toxicity or Zn toxicity. Net photosynthetic rate, stomatal conductance, transpiration rate and instantaneous carboxylation efficiency increased as P or Zn supply increased. Although increasing P supply reduced the P-use efficiency in photosynthesis (PUEP) and increasing Zn supply reduced the Zn-use efficiency in photosynthesis (ZnUEP), increasing Zn supply at a given P supply increased PUEP and increasing P supply at a given Zn supply increased ZnUEP. These results suggest that agricultural management strategies should seek for balanced mineral nutrition to optimize yields and resource-use efficiencies., (© 2018 Scandinavian Plant Physiology Society.)

Cadmium (Cd) was shown to co-localise with calcium (Ca) in oxalate crystals in the stems and leaves of Cd tolerant Gomphrena claussenii, but Cd binding remained unresolved. Using synchrotron radiation X-ray absorption near edge spectroscopy we demonstrate that in oxalate crystals of hydroponically grown G. claussenii the vast majority of Cd is bound to oxygen ligands in oxalate crystals (>88%; Cd-O-C coordination) and the remaining Cd is bound to sulphur ligands (Cd-S-C coordination). Cadmium binding to oxalate does not depend on the amount of Ca supplied or from which organs the crystals originate (stems and mature leaves). By contrast, roots contain no oxalate crystals and therein Cd is bound predominantly by S ligands. The potential to remove Cd by extraction of Cd-rich oxalate crystals from plant material should be tested in phytoextraction or phytomining strategies.

Genetic biofortification requires knowledge on natural variation and the underlying mechanisms of micronutrient accumulation. We therefore studied diversity in grain micronutrient concentrations and spatial distribution in barley (Hordeum vulgare), a genetically tractable model cereal and an important crop with widespread cultivation. We assembled a diverse collection of barley cultivars and landraces and analysed grain micronutrient profiles in genebank material and after three independent cultivations. Lines with contrasting grain zinc (Zn) accumulation were selected for in-depth analysis of micronutrient distribution within the grain by micro-proton-induced X-ray emission (μ-PIXE). Also, we addressed association with grain cadmium (Cd) accumulation. The analysis of > 120 lines revealed substantial variation, especially in grain Zn concentrations. A large fraction of this variation is due to genetic differences. Grain dissection and μ-PIXE analysis of contrasting lines showed that differences in grain Zn accumulation apply to all parts of the grain including the endosperm. Cd concentrations exceeded the Codex Alimentarius threshold in most of the representative barley lines after cultivation in a Cd-contaminated agricultural soil. Two important conclusions for biofortification are: first, high-Zn grains contain more Zn also in the consumed parts of the grain; and second, higher micronutrient concentrations are strongly associated with higher Cd accumulation., (© 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.)

In the last few decades, research has increasingly been aimed at clarifying how root system architecture, physiology and function are related to environmental drivers. 'Negative halotropism' has been defined as the alteration of root growth direction to avoid salinity. We suggested that 'positive halotropism' may be found in halophytes relying on salinity for optimal growth. Investigating root structure of the halophyte Bassia indica (Wight) A. J. Scott, we have shown that positive halotropism can explain the growth of horizontal roots towards optimal salt concentrations along a soil salinity gradient. Here we tested three hypotheses. First, that development of B. indica roots depends on a trade-off between optimal nutrient supply and saline concentrations: results of split-root-experiment showed a preference for sand enriched with nutrients and poor in salts. Second, that shallow horizontal roots enable B. indica to forage for nutrient-rich patches. Results demonstrated that bulk elemental analysis was not consistent with tissue-specific elemental analysis, and this can be explained by substantial variability of element composition of particular root segments. Third, we hypothesised that B. indica redistributes water horizontally through shallow horizontal roots. Results showed that back flow of water from the tap root towards tip root was possible in horizontal roots in saline microenvironment.