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Background: Vitamin D deficiency remains a widespread public health issue in Europe, despite the availability of sunlight, dietary sources, supplements, and food fortification. National fortification strategies differ substantially in their regulatory approaches, food vehicles, and fortification levels, influencing the population’s vitamin D intake and status. Objective: The primary objective of this study was to map vitamin D food fortification policies across European Union (EU) Member States, European Free Trade Association (EFTA) countries, and the United Kingdom (UK), focusing on regulatory frameworks, eligible food categories, and implementation models. Methods: A structured review of national legislation and official guidance on vitamin D food fortification was conducted between December 2025 and March 2026 across EU Member States (n = 27), EFTA countries (n = 4), and the UK. For EU Member States, the framework established by Regulation (EC) No 1925/2006 was examined alongside national implementation measures. For EFTA countries and the UK, corresponding national legislation and official regulatory guidance were reviewed. Data were extracted on fortification policy status, eligible food categories, legal basis, and fortification levels. Targeted searches of PubMed and Scopus were performed to identify modeling studies and policy analyses supporting the interpretation of the findings. Results: Fortification policies show marked heterogeneity. Mandatory fortification is limited to a few countries and specific foods: Finland (homogenized skim milk), Sweden (low-fat milk, fermented dairy, plant-based alternatives, and fat spreads), Belgium (margarine and selected fats), and Poland (margarine and fat spreads). In most other European countries, vitamin D fortification is voluntary under EU legislation or equivalent national legislation, depending on market uptake. Food vehicles vary regionally, with Northern Europe extending fortification beyond fats to include fluid milk and plant-based drinks, whereas other regions mainly fortify margarines, cereals, dairy products, and plant-based beverages. Fortification levels also differ, with some countries specifying maximal or exact levels, while others lack national standards. Data on fortified foods are limited in several Central and Southern European countries. Modeling indicates that multi-vehicle fortification is more effective than single-vehicle approaches, safely increasing population intakes while reducing deficiency prevalence. Conclusions: Vitamin D fortification policies across Europe are highly heterogeneous. Most countries rely on voluntary approaches, which provide limited coverage. Strengthening policy through mandatory and well-coordinated multi-vehicle strategies, informed by modeling and population-based studies, can improve vitamin D intake, reduce deficiency prevalence, and enhance health equity. Full article

Rapid expansion of the plant-based milk market has increased the need to understand how the aroma profiles of these alternatives differ from that of dairy milk and how raw material selection and processing influence volatile formation. This study compared the volatile profiles of dairy milk, commercial plant-based milks, and laboratory-prepared cereal and pseudocereal milk prototypes to identify promising materials for plant-based milk development. Comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOFMS) combined with chemometric analysis was used to characterize volatile compounds in bovine milk, four commercial plant milks, and five laboratory-prepared plant milks. Dairy milk was characterized by fatty acids, esters, and other lipid-derived volatiles, whereas plant-based samples were associated with hydrocarbons, pyrazines, ketones, and phenols. Within the plant-based group, volatile differences were influenced by raw material type and processing history. Commercial products showed more evident processing-related features, whereas laboratory-prepared cereal samples exhibited a simpler volatile background. Among them, barley milk displayed a distinctive toasted and cereal-like signature. Overall, the selected cereal and pseudocereal matrices showed distinct volatile characteristics, as well as relatively uniform raw material backgrounds, implying greater flexibility in aroma expression. These features make them promising candidates for dairy alternatives and may help guide future plant-based milk formulation. Full article

Cultivated lettuce (Lactuca sativa L.) is a major leafy crop and an emerging model for functional genomics within the Asteraceae family, supported by high-quality reference genomes and efficient transformation systems. Although CRISPR/Cas technology offers powerful opportunities for crop improvement, editing efficiency depends on optimized construct architecture and reliable guide RNA (gRNA) validation. However, a rapid platform for evaluating CRISPR reagents in lettuce is still lacking. Here, we developed an efficient hairyroot-based system to accelerate CRISPR/Cas genome editing optimization in L. sativa. Four Agrobacterium rhizogenes strains were compared for hairy root induction in two cultivars, ‘Saladin’ and ‘Osiride’, identifying strain ATCC15834 as the most effective based on transformation frequency and root production. Using this platform, we evaluated multiple CRISPR construct configurations, including alternative promoters for nuclease and gRNA expression. A plant-derived promoter combined with At-pU6-26 variant significantly improved editing efficiency. As a proof of concept, we targeted LsHB2, the putative ortholog of Arabidopsis thaliana ATHB2, a key regulator of the shade avoidance response using SpCas9, SaCas9, and LbCas12a nucleases. The system enabled rapid genotyping and quantitative indel profiling. Overall, this workflow provides a robust framework for efficient guide selection and construct optimization in lettuce genome editing. Full article

One of the main goals of heat and electricity producers in Latvia is to reduce the use of fossil fuels and introduce alternative fuel types that could help in reducing carbon dioxide emissions. This work focuses on addressing the set issue for a medium-capacity automated gas boiler plant, which provides heat for a local residential district. The following solutions were selected for boiler plant optimization: an electric boiler, a heat storage system, and solar collectors. Operating mode simulations were conducted for the electric boiler and solar collectors using Excel and Polysun (Standard) software. Simulations were created based on energy resource demand data obtained from a residential district located in Latvia and local energy resource prices/heat energy tariffs for the year 2024. The results from the simulations were used for technical and economic calculations to determine the payback period of the project. The electric boiler, together with the thermal energy storage tank and solar collectors, can produce 5903.04 MWh/year (~70% of local district heat demand) of thermal energy. This reduces the CO₂ emissions of the boiler plant by at least 1186.51 tCO₂ per year, which, at an emission quota price of 63.80 EUR/tCO₂, allows for savings of 75,699.34 EUR per year (12.82 EUR/MWh heat energy). The project’s discounted payback period is 4.12 years, considering the reduction in the cost of the CO₂ emission quota. The results of this study show that the chosen technologies are straightforward solutions that can be used to optimize existing boiler plants with limited space and can provide financial benefits to heat energy producers. Full article

Microbial oil production has gained attention as a sustainable and cost-effective alternative to conventional vegetable and fish oils. Among oleaginous microorganisms, Yarrowia lipolytica is notable for its ability to accumulate lipids exceeding 20% of its dry weight. This study aimed to evaluate semi-continuous cultivation as a strategy for sustainable microbial oil production by Y. lipolytica, while also assessing the potential utility of the spent supernatant. Three different feeding frequencies were evaluated. In the 24 h feeding regime, the maximum oil concentration reached 11.22 g/L, decreasing to 8.43 g/L by the 88th hour. In the 6–6–12 h feeding strategy, crude protein content peaked at 43.75% of dry mass at 22 h. Fatty acid profiling revealed consistently low saturated fatty acid (SFA) levels (4.93–10.30%), while unsaturated fatty acids (UFA) dominated (89.69–95.05%). Monounsaturated fatty acids (MUFA) were predominant, reaching up to 81.24%, whereas polyunsaturated fatty acids (PUFA) ranged from 20.78% to 29.98%. Oleic acid was the most abundant fatty acid across all conditions. This composition supports the potential of microbial oil from Y. lipolytica as a sustainable alternative edible lipid ingredient for human food applications, complementing conventional plant-based oils. The favorable unsaturated fatty acid profile indicates its potential suitability for incorporation into food formulations requiring nutritionally desirable lipid sources. As part of the sustainability-oriented approach of the study, the freeze-dried post-culture supernatant was also evaluated for its potential further utilization. With a calorific value of 10.43 kJ/g and significant phosphorus and potassium levels, it shows potential as a biofuel feedstock and as a biofertilizer or biostimulant. Full article

Compost supplementation is widely used to improve yield and crop consistency in the cultivation of white button mushroom (Agaricus bisporus), yet practical alternatives to conventional protein-rich supplements and rapid candidate-screening approaches are still needed. In this study, plant- and byproduct-based supplements were first compared by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) to obtain qualitative fingerprints of extractable protein fractions, and were then evaluated in Phase III cultivation under both bag-based screening conditions and in a large-scale pull-mat system. Supplements differed notably in protein banding patterns and cultivation performance. In the bag trials, lupin grist and corn pellet produced the largest yield increases relative to the non-supplemented control, whereas in the commercial pull-mat trials lupin grist was the best-performing supplement, reaching 240.77 kg t⁻¹ compost. Under the present conditions, SDS-PAGE was useful as a qualitative screening aid for prioritizing candidates for cultivation trials, but not as a stand-alone predictor of yield. These results identify lupin grist as a practically relevant supplement candidate for commercial A. bisporus production. Full article

Cancer remains a leading cause of morbidity and mortality globally, with current therapies often limited by toxicity, drug resistance, and reduced efficacy in advanced stages. Medicinal plants represent important sources of bioactive compounds (BACs) with anticancer and chemopreventive potential; however, their successful application is strongly influenced by extraction strategies that determine phytochemical recovery and downstream biological activity. This review evaluates solvent-based extraction techniques used to extract BACs from medicinal plants with reported anticancer properties, synthesizing peer-reviewed articles from PubMed and Google Scholar published between 2020 and 2025. Solvent-based methods, including Soxhlet and maceration, were most widely applied due to their operational simplicity and the preservation of structurally diverse metabolites while percolation, decoction, infusion, and hydro-distillation were sparsely utilized. Extraction strategy and solvent polarity emerged as primary factors shaping phytochemical profiles, with phenolics, flavonoids, alkaloids, and terpenoids identified as dominant classes. Reported half maximal inhibitory concentration (IC₅₀) ranged from highly potent (0.12 µg/mL) to weak (30,000 µg/mL), reflecting variability driven by extraction parameters and plant matrix complexity. Anticancer mechanisms commonly involved apoptosis induction, cell-cycle arrest, reactive oxygen species-mediated cytotoxicity, and inhibition of proliferative signaling pathways across breast, cervical, colon, lung, liver, and prostate cancer models. Although solvent-based extraction approaches remain widely used, their context-dependent nature and lack of standardization limit reproducibility. Overall, anticancer and chemotherapeutic efficacy is primarily governed by BAC composition, while extraction methods act as upstream modulators. Future progress requires phytochemical-informed, standardized workflows supported by hybrid extraction systems, AI-assisted optimization, and advanced bioavailability and delivery systems to enable reproducible and clinically relevant translation of plant-derived chemotherapeutics. Full article

Efficient water management is essential for sustainable agriculture, particularly for crops like carrots that are traditionally grown in soil systems with high water consumption, averaging 4500–6000 m³/ha. Hydroponics offers a potential alternative due to its higher water-use efficiency, yet root crops have been understudied because of system design and economic challenges. This study evaluated the effects of different hydroponic solution volumes on the growth of carrots (Daucus carota cv. Mokum) in a Deep-Water Culture (DWC) system to address knowledge gaps regarding their feasibility in soilless production. Experiments were conducted in a controlled greenhouse using three solution volume treatments (50% with 10.75 L, 75% with 16.13 L, and 100% with 21.50 L) applied to 12 plants per treatment across two repeated experiments. Biomass production, water use efficiency, and total carotenoid concentration were assessed after eight weeks. The 100% (21.50 L) volume treatment produced the greatest shoot and root biomass, whereas the 50% (10.75 L) volume treatment significantly increased total carotenoid concentration, particularly in the second trial. Despite lower water inputs, water use efficiency did not differ statistically among treatments. These results indicate that carrots can be successfully cultivated in DWC systems, though further optimization, such as using narrower containers, may be required to improve efficiency and competitiveness with soil-based production. Full article

Energy recovery from sewage sludge represents a sustainable and technically feasible alternative to promote integration between environmental sanitation and renewable energy generation. This study presents a case analysis of the municipality of Ribeirão Preto, São Paulo, focusing on comparisons between two wastewater treatment systems: an Upflow Anaerobic Sludge Blanket (UASB) reactor and a continuous-flow activated sludge system. Using the UASB configuration, we prepared a preliminary design of a treatment plant based on population and effluent generation projections over a 20-year horizon. The estimated sludge and biogas production allowed us to simulate electricity generation then. The comparative economic assessment, which employed Net Present Value (NPV) and Internal Rate of Return (IRR) indicators in accordance with ANEEL Resolution No. 482/2012, showed that the UASB system yields hard superior methane (up to 3235.6 m³/day) and higher electricity generation potential (1839.7 MWh/year) than the activated sludge system (1990 m³/day and 1654.3 MWh/year, respectively). Both systems were economically viable, with a positive NPV, an IRR of up to 16.83%, and payback periods starting in the first cycle. Furthermore, we estimated the cost per cubic meter of generated biomethane, conducted a sensitivity analysis, and assessed the impact on the most important economic indicators, all to identify the advantages and disadvantages of the proposed project and the best use of the generated biogas. This analysis showed that it is possible to recover energy from sewage treatment systems while also reusing sewage sludge for agricultural applications, thereby highlighting additional environmental and economic benefits, particularly in regions with a strong presence of agribusiness, e.g., Ribeirão Preto. Full article

The use of treated wastewater constitutes a strategic alternative for agriculture in water-scarce regions. This study developed and applied a distance-based and sodium-constrained deterministic allocation model integrating geoprocessing tools with environmental and logistical constraints to optimize the spatial distribution of treated effluent from 48 wastewater treatment plants (WWTPs) in the semi-arid region of Minas Gerais, Brazil. The deterministic allocation algorithm prioritizes geographic proximity and favorable topographic differences as a proxy for reducing potential pumping requirements. Two scenarios were evaluated: (1) full effluent availability and (2) sodium-regulated allocation limited to 300 kg ha⁻¹ year⁻¹ of Na, in accordance with Normative Deliberation CERH-MG 65/2020. Under Scenario 1, cotton demand exceeded (184%), while coffee and sugarcane reached 69% and 24% of annual demand, respectively. Under the sodium-constrained Scenario 2, demand fulfillment changed to 37% for coffee and 42% for sugarcane, while cotton remained above full demand (108%). The proposed model differs from previous deterministic spatial allocation applications by integrating regulatory sodium constraints and dual-scenario regional assessment, providing a spatially explicit and regulation-compliant decision-support tool for sustainable wastewater reuse in semi-arid agricultural systems. Full article

Amid growing environmental concerns regarding the use of non-biodegradable plastic packaging and its potential emerging contaminants, such as microplastics, currently among the most pressing global challenges, researchers in the food sector are increasingly pursuing sustainable alternatives. In this context, various organic sources have been explored for the development of innovative biocompatible films. These films exhibit properties such as low water vapor permeability, transparency, and biodegradability, and have recently gained active functionalities. These enable the extension of the shelf life of packaged foods by controlling microbial activity and oxidative degradation. Lipid-based compounds derived from animal and plant sources—including phospholipids, essential oils, free fatty acids, and saturated and polyunsaturated fatty acids—have proven highly effective when incorporated into films, leading to significant physicochemical, mechanical, and microbiological improvements in both the films and the packaged products. Owing to their high hydrophobic capacity, these lipids markedly reduce water vapor permeability, which is crucial for extending the shelf life of high-moisture foods. Studies have shown that the incorporation of lipid compounds can increase film tensile strength by up to 37% and enhance antioxidant activity by over 75%. Moreover, many of these compounds exhibit antibacterial and antimicrobial activities, becoming active on the surface of food in contact. However, many bioactive compounds have poor dispersion in aqueous solutions, limiting their effectiveness in the final product. When encapsulated with the aid of a lipid fraction, the bioavailability of these compounds is improved, and their release can be effectively controlled. This review aims to consolidate recent research on the production of biopolymer films incorporating various types of lipid compounds, highlighting their enhancements and potential applications in active food packaging systems. Full article

In gasification-based waste treatment plants, continuous generation of combustible gas is essential for stable and efficient operation. To achieve this, multiple gasification furnaces are operated alternately; however, the internal states of the furnaces cannot be directly observed, making it difficult to assess the progress of gasification. Consequently, operation planning relies heavily on the experience of skilled operators. In this study, nonlinear system identification models based on deep learning are developed to predict the valve opening that controls the injection of gasification agents, which implicitly reflects the gasification state. Several modeling approaches, including linear finite impulse response (FIR) models, block-oriented Hammerstein–Wiener (HW) models, deep Hammerstein–Wiener models, and Transformer-based models, are investigated and compared. The models are trained and validated using actual operational data obtained from an industrial waste treatment plant. The results demonstrate that nonlinear models significantly outperform linear models, particularly for long-term prediction horizons. Among the examined approaches, the Transformer-based model shows stable and competitive performance across different prediction intervals. These findings indicate that deep learning-based nonlinear modeling is effective for predicting plant operation and has the potential to support automated operation planning, thereby reducing reliance on operator expertise. Full article

The excessive use of chemical fertilizers in rice cultivation has contributed to soil degradation, creating a need for sustainable biological alternatives. This study examined the functional diversity of three indigenous nostocalean cyanobacterial strains (UP1, UP2, and UP3) isolated from forest and paddy field ecosystems by comparing the effects of their cellular biomass and extracellular polymeric substances (EPS) on rice seedling growth and soil properties. Morphological observations and partial 16S rRNA sequence analysis indicated that strains UP1 and UP2 were affiliated with the genus Ahomia, whereas UP3 was placed within the genus Nostoc. Together, these results placed all three isolates within the heterocystous cyanobacterial order Nostocales. The strains were further characterized based on EPS production and its degree of polymerization. Seed germination and seedling vigor assays were conducted to select the most effective biomass and EPS treatments, which were subsequently evaluated in 21-day pot experiments. Fresh biomass from strain UP2 most effectively enhanced rice growth, whereas EPS from strain UP3 promoted root development. EPS application from strain UP3 significantly increased root elongation to 13.44 cm, while high biomass levels of UP2 increased total sugar and free amino acid contents, indicating distinct plant response patterns. Soil analyses revealed differential responses between biomass- and EPS-based applications, with biomass generally producing stronger effects. Biomass from all strains was associated with higher physical soil function index (PSFI) values (up to 1.35). In contrast, improvements in chemical soil function index (CSFI) were observed across treatments, with variable responses and relatively higher values recorded in biomass from strain UP3 (up to 1.24). These findings suggest strain- and form-dependent response patterns of nostocalean cyanobacteria with potential for enhancing rice growth and improving soil functionality under the controlled conditions. Full article

Leaf Area Index (LAI) is critical for monitoring cassava growth and yield prediction, yet ground measurements are time-consuming and labor-intensive for large-scale applications. While satellite-based vegetation indices (VIs) offer a scalable alternative, their performance for cassava LAI estimation remains poorly documented, and optimal index selection for different growth stages is unclear. This study evaluated the predictive performance of 13 Sentinel-2-derived VIs for estimating ground-measured LAI across cassava growth stages. Ground-LAI was measured monthly using a SunScan Canopy Analyzer from January to June 2022 (2–7 months after planting; MAP) in 47 cassava plots in Nakhon Ratchasima Province, Thailand. Linear mixed-effects models and stage-specific regressions assessed VI predictive performance using Coefficient of determination (R²) and Root Mean Squared Error (RMSE). The Green Normalized Difference Vegetation Index (GNDVI) and Normalized Difference Water Index (NDWI) demonstrated superior performance across all growth stages (R² = 0.524; RMSE = 0.350), followed by Sentinel-2 LAI Green Index (SeLI R² = 0.521, RMSE = 0.357). Stage-specific analysis revealed that Ratio Vegetation Index performed best during early growth (2 MAP, R² = 0.671; RMSE = 0.164) while GNDVI and NDWI excelled during mid-growth (3–5 MAP) and SeLI at late growth (7 MAP, R² = 0.393; RMSE = 0.422). While the presence of large trees altered the ranking of VI predictive performance, it did not substantially affect estimation errors, suggesting a relatively small impact of spatial heterogeneity on LAI estimation accuracy. These findings identify GNDVI and NDWI as the most operationally suitable Sentinel-2 indices for cassava LAI estimation and demonstrate that stage-specific index selection can improve monitoring accuracy, providing validated tools for regional-scale cassava crop monitoring using freely available satellite data. Full article

Epithelial ovarian carcinoma (EOC) is the most common type of ovarian cancer and represents the most lethal gynecologic neoplasm. EOC is usually diagnosed at late stages due to its nonspecific signs and symptoms. Although significant clinical advances have been made in other types of malignancies, EOC remains a disease that requires further biological research to identify new therapeutic targets or new treatment alternatives, as conventional approaches are often ineffective or lead to the development of resistance and unwanted side effects. There are a significant number of natural products from which commercially available drugs have been derived, largely for the treatment of cancer, but none of them focus on epigenetic changes in specific targets in EOC. Based on the above, this work focuses on describing the in vitro and in vivo findings from the last twelve years derived from the action of important phytochemicals on epigenetic targets in ovarian cancer, among other mechanisms of action, revealing that there is a significant gap to be bridged in terms of the transition from basic to applied research regarding the potential of plant-derived molecules as possible epidrugs in EOC. Full article