The concept of functional nutrition has garnered mounting attention, primarily due to growing evidence that specific dietary components have the capacity to provide health benefits that extend beyond the mere supply of basic nutrients. In this context, glucosinolate-rich species of the Brassicales order are of importance as a source of bioactive compounds, which exhibit antioxidant, anti-inflammatory, and chemoprotective properties. The review identifies which Brassicales species may be considered as functional foods or functional ingredients. It does so by starting from their glucosinolate profile, summarizing their potential applications in disease prevention, and highlighting current strategies aimed at enhancing glucosinolate levels through agronomic practices and processing approaches. The potential applications of the main species of the Brassicales order in the prevention of cardiovascular, obesity-related and degenerative diseases, as well as in the development of functional foods, are highlighted. These species are considered both as ready-to-use functional foods and as functional ingredients that can be obtained through extraction or fermentation processes, including the valorization of agricultural waste.

Colony losses in Apis mellifera are caused by multiple stressors, among which the parasitic microsporidian Nosema (= Vairimorpha) ceranae. As an alternative to the controversial fumagillin, sustainable control strategies based on natural bioactive compounds are increasingly being explored. This study evaluated the efficacy of glucosinolate (GSL)-based formulations derived from flowers of Eruca sativa and Reseda lutea in controlling N. ceranae infection in worker honey bees under laboratory conditions. Freeze-dried flowers were characterized for their GSL profile and myrosinase activity, and formulation studies were conducted to choose the best material for bioassays. Then, water-soluble GSL-enriched extracts were produced. Cage feeding assays were performed on uninfected newly emerged bees using sugar syrup supplemented with 2 and 4 µmol g⁻¹ of total GSL of either formulation. Extract administration reduced food consumption and bee survival in a concentration-dependent manner compared to the control group. The lower concentration was further tested on 6-day-old bees artificially infected with N. ceranae spores, showing consumption comparable to control, but still a slight decrease in survival. At this concentration, both E. sativa- and R. lutea-based extracts significantly inhibited N. ceranae abundance, by 23.8 and 44.6

Glucosinolates are secondary metabolites of the Brassicales, playing a role in plant protection and as health-promoting compounds. Here, Na2SO4 was used to modulate the aliphatic glucosinolate content in different organs of Eruca sativa Mill. In flowers, which accumulate the highest amount of glucosinolates, Na2SO4 increased the concentration of glucoraphanin, in roots of glucoerucin and in apical leaves it doubled the amount of dimeric 4-mercaptobutyl glucosinolate. The biosynthetic gene Branched-Chain Aminotransferase 4 was also induced in roots at the highest salt concentration, while in leaves all tested genes biosynthetic genes were downregulated or unaffected. Cytochromes P450 83A1 monooxygenase was downregulated at the highest salt concentration in all organs. Overall, E. sativa is a reliable source of glucosinolates, which can be modulated with Na2SO4.

Background/Objectives: Crude extracts from the Brassica genus have recently emerged as promising phytochemicals for preventing bone loss. While the most documented evidence suggests that their general biological activity is due to glucosinolates’ (GLSs’) hydrolysis products, the direct activity of GLSs is beginning to be uncovered. However, the contribution of GLSs to the bone-sparing activity of crude Brassicaceae extracts has seldom been addressed. Here, we aimed to gain insights into this gap by studying in the same in vitro model of human osteogenesis the effect of two Brassica seed extracts (Eruca sativa and Lepidium sativum) obtained from defatted seed meals, comparing them to the isolated GLSs most represented in their composition, glucoerucin (GER) and glucotropaeolin (GTL), for Eruca sativa and Lepidium sativum, respectively. Methods: Osteogenic differentiation of human mesenchymal stromal cells (hMSCs) was assessed by alizarin red staining assay and real-time PCR, respectively, evaluating mineral apposition and mRNA expression of specific osteogenic genes. Results: Both Brassica extracts and GLSs increased the osteogenic differentiation, indicating that the stimulating effect of Brassica extracts can be at least partially attributed to GLSs. Moreover, these data extend previous evidence of the effect of unhydrolyzed glucoraphanin (GRA) on osteogenesis to other types of GLSs: GER and GTL. Notably, E. sativa extract and GTL induced higher osteogenic stimulation than Lepidium sativum extract and GER, respectively. Conclusions: Overall, this study expands the knowledge on the possible application of Brassica-derived bioactive molecules as natural alternatives for the prevention and treatment of bone-loss pathologies.

Crambe abyssinica Hochst defatted seed meals were used to produce protein hydrolysates through a mild enzymatic two-step hydrolysis process. The resulting hydrolysates were rich in free amino acids, low-molecular-weight peptides, and potential bioactive compounds such as phenols, glucosinolates, or their derivatives. These hydrolysates were tested in bioassays, performed under controlled conditions, on mung bean (Vigna radiata) cuttings, to investigate a possible auxin effect, and on maize (Zea mays L.) in an aeroponic/hydroponic system during the first two weeks of growth. In both assays, crambe hydrolysates revealed a stimulating effect on root development at a dose corresponding to nitrogen concentration of 4.8 mM, promoting lateral root formation and altering root architecture. Furthermore, they exhibited a positive impact on nitrogen content in both maize roots and shoots, along with an increase in the chlorophyll SPAD index. Notably, the observed effects were similar to those induced by a commercial biostimulant based on an animal-derived hydrolysate, tested under the same conditions on maize. The present work underscores the potential of crambe seed by-products for new sustainable and environmentally safe agro-inputs aimed at enhancing crop performance within the framework of a circular economy.

Safflower (Carthamus tinctorius L.) is an underestimated and multipurpose crop resistant to environmental stresses. Its oil presents useful chemical–physical properties, potentially exploitable for industrial purposes as a bio-based lubricant. In this work safflower oil was applied as a less toxic alternative to mineral-based hydraulic fluids. The extracted oil was partially refined and the antioxidant tert-buthylhydroquinone (THBQ) was added at two concentrations (0.25 and 3.00 mg kg−1). Efficiency tests of the obtained oil were carried out using an experimental test rig capable of simulating a real hydraulic system and performing severe short-duration work cycles with the aim of strongly accelerating the ageing of the tested oil. Oil performance was verified by monitoring hydraulic and chemical–physical parameters, which were correlated to the main lubricant properties through sensor detection and laboratory analysis in parallel. The results indicated that the safflower oil behaved well at both THBQ concentrations and showed good technical performance (operating pressure and temperature; flowrate and transmitted hydraulic power), though a higher THBQ concentration was necessary to protect the oil’s chemical–physical properties from worsening. In fact, the higher THBQ concentration allowed the test to be extended to 270 h, an improvement compared to the 150 h that was achieved with the lower THBQ concentration. Finally, the use of safflower oil for industrial and agricultural purposes seems feasible and would contribute toward the sustainability of the whole crop rotation in a prospective valuable circular economy.

Modulation of mitochondrial K channels represents a pharmacological strategy to promote cardioprotective effects. Isothiocyanates emerge as molecules capable of releasing hydrogen sulfide (H2S), an endogenous pleiotropic gasotransmitter responsible for anti-ischemic cardioprotective effects also through the involvement of mitoK channels. Erucin (ERU) is a natural isothiocyanate resulting from the enzymatic hydrolysis of glucosinolates (GSLs) present in Eruca sativa Mill. seeds, an edible plant of the Brassicaceae family. In this experimental work, the specific involvement of mitoKATP channels in the cardioprotective effect induced by ERU was evaluated in detail. An in vivo preclinical model of acute myocardial infarction was reproduced in rats to evaluate the cardioprotective effect of ERU. Diazoxide was used as a reference compound for the modulation of potassium fluxes and 5-hydroxydecanoic acid (5HD) as a selective blocker of KATP channels. Specific investigations on isolated cardiac mitochondria were carried out to evaluate the involvement of mitoKATP channels. The results obtained showed ERU cardioprotective effects against ischemia/reperfusion (I/R) damage through the involvement of mitoKATP channels and the consequent depolarizing effect, which in turn reduced calcium entry and preserved mitochondrial integrity.

BACKGROUND:Hydrogen sulfide (H2S) is a gasotransmitter deeply involved in cardiovascular homeostasis and implicated in the myocardial protection against ischemia/reperfusion. The post-translational persulfidation of cysteine residues has been identified as the mechanism through which H2S regulates a plethora of biological targets. Erucin (ERU) is an isothiocyanate produced upon hydrolysis of the glucosinolate glucoerucin, presents in edible plants of Brassicaceae family, such as Eruca sativa Mill., and it has emerged as a slow and long-lasting H2S-donor.AIM:In this study the cardioprotective profile of ERU has been investigated and the action mechanism explored, focusing on the possible role of the recently identified mitochondrial Kv7.4 (mitoKv7.4) potassium channels.RESULTS:Interestingly, ERU showed to release H2S and concentration-dependently protected H9c2 cells against H2O2-induced oxidative damage. Moreover, in in vivo model of myocardial infarct ERU showed protective effects, reducing the extension of ischemic area, the levels of troponin I and increasing the amount of total AnxA1, as well as co-related inflammatory outcomes. Conversely, the pre-treatment with XE991, a blocker of Kv7.4 channels, abolished them. In isolated cardiac mitochondria ERU exhibited the typical profile of a mitochondrial potassium channels opener, in particular, this isothiocyanate produced a mild depolarization of mitochondrial membrane potential, a reduction of calcium accumulation into the matrix and finally a flow of potassium ions. Finally, mitoKv7.4 channels were persulfidated in ERU-treated mitochondria.CONCLUSIONS:ERU modulates the cardiac mitoKv7.4 channels and this mechanism may be relevant for cardioprotective effects.

Eruca sativa Miller (Brassicaceae) is an insect-pollinated diploid annual species which grows spontaneously in the entire Mediterranean basin from semi-arid to arid-hot conditions and is cultivated in Northern America, Europe, and Asia as either salad or oilseed crop. Here, some essential background was provided on this versatile crop, summarizing the present status of Eruca sativa research focusing on the wealth of bioactive ingredients in its seeds, which may find exploitation in agriculture, in the food industries and as nutraceuticals for their antioxidant and anti-inflammatory properties. Fatty acids of Eruca sativa seed oil, gums, glucosinolates and soluble and insoluble phenol and flavonoid fractions in the defatted press cake are the main bioactive compounds considered to date by the scientific literature and that deserve attention for their physical and biological activities.

Introduction:Most patients with metabolic syndrome present non-alcoholic fatty liver disease (NAFD).High consumption of fruit and vegetable has been related to a lower incidence of chronic diseases.Of note, among vegetables, brassicaceae family is at the first place.Therapeutic strategies to treat inflammatory disease as well as cancer are evolving and includes biologically active phytocompounds.A sulfur-containing glucosinolate (GLS) secondary plant compounds exclusively present in Brassicaceae-and their breakdown products including isothiocyanates (NCS) have shown to possess important anti-inflammatory and anti-carcinogenic property.The aim of this work was the evaluation of the effects of daily consumption of E. sativa defatted seed meal enriched bakery products on glucose and lipid metabolism and on systemic markers of inflammation. Material and Methods:After an 8-week run-in period, patients were asked for 4 weeks to maintain their usual diet replacing 150 gr. of bakery products with the same number of enriched bakery product.Fasting glucose and insulin, HOMA-r, total cholesterol, HDL, LDL, triglyceride, ALT, Gamma-GT, high sensitivity C reactive protein, Interleukin-6, Interleukin-8 and TNF-α were compared before and after the study period.Results: HOMA-r showed a statistically significant reduction after 4 weeks period as well as cholesterol ratio 3.8 ± 1.0 vs. 3.6 ± 1.0 (P = 0.03).Among markers of inflammation, high sensitivity CRP showed a statistically significant reduction (-36.8%)from 1.4 (0.4 -10.4) to 0.9 (0.4 -8.7) mg/dl (P = 0.02) as well as TNF-α 8.0 ± 3.0 vs. 7.1 ± 2.7 pg/ml (-11.3%;P < 0.001).Despite a mild ALT reduction (-7.0%),Gamma-GT showed a -21.0%decrease from 20.0 ± 6.6 to 15.8 ± 6.4 U/L (P < 0.001).IL-6 and IL-8 were substantially unmodified. Conclusion:as cracker can improve glucose and lipids metabolism in parallel with a significant improvement of serum Bakery products enriched with Eruca sativa Mill defatted seed meal markers of inflammation and therefore being of interest for patients affected by inflammatory diseases.

Nosema ceranae is a widespread parasite responsible for nosemosis Type C in Apis mellifera honey bees, reducing colony survival. The antibiotic fumagillin is the only commercial treatment available, but concerns are emerging about its persistence, safety, and pathogen resistance. The use of natural substances from Brassicaceae defatted seed meals (DSMs) with known antimicrobial and antioxidant properties was explored. Artificially infected bees were fed for 8 days with candies enriched with two concentrations, 2% and 4%, of two DSMs from Brassica nigra and Eruca sativa, containing a known amount of different glucosinolates (GSLs). The food palatability, GSL intake, bee survival, and treatment effects on N. ceranae spore counts were evaluated. Food consumption was higher for the two 2% DSM patties, for both B. nigra and E. sativa, but the GSL intake did not increase by increasing DSM to 4%, due to the resulting lower palatability. The 2% B. nigra patty decreased the bee mortality, while the higher concentration had a toxic effect. The N. ceranae control was significant for all formulates with respect to the untreated control (312,192.6 +/− 14,443.4 s.e.), and was higher for 4% B. nigra (120,366.3 +/− 13,307.1 s.e.). GSL hydrolysis products, the isothiocyanates, were detected and quantified in bee gut tissues. Brassicaceae DSMs showed promising results for their nutraceutical and protective effects on bees artificially infected with N. ceranae spores at the laboratory level. Trials in the field should confirm these results.

The microsporidian fungus Nosema ceranae represents one of the primary bee infection threats worldwide and the antibiotic fumagillin is the only registered product for nosemosis disease control, while few alternatives are, at present, available. Natural bioactive compounds deriving from the glucosinolate–myrosinase system (GSL–MYR) in Brassicaceae plants, mainly isothiocyanates (ITCs), are known for their antimicrobial activity against numerous pathogens and for their health-protective effects in humans. This work explored the use of Brassica nigra and Eruca sativa defatted seed meal (DSM) GSL-containing diets against natural Nosema infection in Apis mellifera colonies. DSM patties from each plant species were obtained by adding DSMs to sugar candy at the concentration of 4% (w/w). The feeding was administered in May to mildly N. ceranae-infected honey bee colonies for four weeks at the dose of 250 g/week. In the treated groups, no significant effects on colony development and bee mortality were observed compared to the negative controls. The N. ceranae abundance showed a slight but significant decrease. Furthermore, the GSL metabolism in bees was investigated, and MYR hydrolytic activity was qualitatively searched in isolated bee midgut and hindgut. Interestingly, MYR activity was detected both in the bees fed DSMs and in the control group where the bees did not receive DSMs. In parallel, ITCs were found in gut tissues from the bees treated with DSMs, corroborating the presence of a MYR-like enzyme capable of hydrolyzing ingested GSLs. On the other hand, GSLs and other GSL hydrolysis products other than ITCs, such as nitriles, were found in honey produced by the treated bees, potentially increasing the health value of the final product for human consumption. The results are indicative of a specific effect on the N. ceranae infection in managed honey bee colonies depending on the GSL activation within the target organ.

Animal manure application to soils is considered to be one of the main cause of antibiotic and bacterial pathogen spread in the environment. Pig livestock, which is the source of one of the most used fertilizer for cultivated land, is also a hotspot for antibiotics and antibiotic-resistant bacteria. Besides harsh chemical and physical sanitization treatments for the abatement of antibiotics and bacterial load in livestock waste, more sustainable and environmentally friendly strategies need to be considered. In this context, the use of natural substances which are proved useful for pest and disease control is currently under exploration for their role in the reduction of bacterial pathogen population. Among these, plants and derived products from the Brassicaceae family, characterized by the presence of a defensive glucosinolate-myrosinase enzymatic system, have been successfully exploited for years in agriculture using the so-called biofumigation technique against crop diseases. Although the application of biofumigation to suppress a range of soil borne pests has been well documented, no studies have been examined to reduce bacterial population in animal waste. In the present study, the release and the antibacterial activity of bioactive compounds deriving from different Brassicaceae defatted seed meals against pathogens and bacterial population in pig manure is addressed. Rapistrum rugosum and Brassica nigra defatted seed meals were found to be the most active products against tested pathogens and able to significantly reduce the bacterial load in the manure.

Vegetable oils are well known for their potential applications in green chemistry, including use as hydraulic fluids. Among oilseed crops, Crambe abyssinica Hochst. EX R.E.Fr . gained attention for its low-input requirements during cultivation and for the properties of its oil, characterised by a high erucic acid content. This long-chain mono-unsaturated fatty acid provides appreciable features that make the oil suitable for interesting green chemistry sectors, as biolubricants and cosmetics. In the present work, Crambe oil was tested as a hydraulic fluid for sustainable agricultural applications. Crambe oil was partially refined through phospholipid removal and added with a food-grade antioxidant (tert-butylhydroquinone) at two different concentrations. The fluid efficiency tests were carried out using an experimental test rig, able to simulate a real hydraulic device, performing heavy work cycles at 40-MPa pressure and at 100 °C temperature, with the aim of strongly accelerating the ageing of the tested fluid. At a lower antioxidant concentration, 0.25 g kg−1, the oil underwent a very quick degradation process. However, increasing the additive dose to 2.0 g kg−1, the fluid maintained stable performances. Indeed, all parameters, referred to oil chemical-physical stability and technical performance, were constant along the entire work cycle, up to 290 h. Finally, the present work showed how Crambe seed cultivation, oil extraction and exploitation in the hydraulic circuit of farm machinery could be developed applying green chemistry approaches aiming at small-scale biorefineries linked to the local supply.

Hydrogen peroxide is an unavoidable by-product of cell metabolism, but when it is not properly managed by the body it can lead to several pathologies (e.g., premature aging, cardiovascular and neurodegenerative diseases, cancer). Several methods have been proposed for the measurement of intracellular H2O2 but none of them has proven to be selective. We developed a rapid all-in-one chemiluminescent bioassay for the quantification of H2O2 in living cells with a low limit of detection (0.15 μM). The method relies on an adamantylidene-1,2-dioxetane lipophilic probe containing an arylboronate moiety; upon reaction with H2O2 the arylboronate moiety is converted to the correspondent phenol and the molecule decomposes leading to an excited-state fragment that emits light. The probe has been successfully employed for quantifying intracellular H2O2 in living human endothelial, colon and keratinocyte cells exposed to different pro-oxidant stimuli (i.e., menadione, phorbol myristate acetate and lipopolysaccharide). Imaging experiments clearly localize the chemiluminescence emission inside the cells. Treatment of cells with antioxidant molecules leads to a dose-dependent decrease of intracellular H2O2 levels. As a proof of concept, the bioassay has been used to measure the antioxidant activity of extracts from Brassica juncea wastes, which contain glucosinolates, isothiocyanates and other antioxidant molecules.

Lactic acid bacteria (LAB) "fermentates" confer a beneficial effect on intestinal function. However, the ability of new fermentations to improve LAB broth activity in preventing pathogen-induced intestinal inflammation and barrier dysfunction has not yet been studied. The objective of this study was to determine if broths of LAB fermented with Eruca sativa or Barbarea verna seed extracts prevent gut barrier dysfunction and interleukin-8 (CXCL8) release in vitro in human intestinal Caco-2 cells infected with enterohemorrhagic Escherichia coli (EHEC) O157:H7. LAB broths were assayed for their effects on EHEC growth and on Caco-2 viability; thereafter, their biological properties were analysed in a co-culture system consisting of EHEC and Caco-2 cells. Caco-2 cells infected with EHEC significantly increased CXCL8 release, and decreased Trans-Epithelial Electrical Resistance (TEER), a barrier-integrity marker. Notably, when Caco-2 cells were treated with LAB broth enriched with E. sativa seed extract and thereafter infected, both CXCL8 expression and epithelial dysfunction reduced compared to in untreated cells. These results underline the beneficial effect of broths from LAB fermented with E. sativa seed extracts in gut barrier and inflammation after EHEC infection and reveal that these LAB broths can be used as functional bioactive compounds to regulate intestinal function.

Background and PurposeHydrogen sulfide (H2S)‐releasing agents are viewed as potential antihypertensive drugs. Recently, natural isothiocyanates emerged as original H2S‐donor agents. Among them, erucin, present in some edible cruciferous plants, shows suitable H2S‐releasing properties and features of “druggability.” The aim of this work was to investigate the erucin‐mediated release of H2S inside vascular cells, its vasorelaxing effects, and activity on BP of normo and hypertensive animals.Experimental ApproachIntracellular H2S‐release and the hyperpolarizing effect of erucin were tested using fluorescent dye, in human aortic smooth muscle cells (HASMCs). Its direct vasorelaxing effect and ability to inhibit noradrenaline‐induced vasoconstriction were evaluated on endothelium‐intact or ‐denuded rat aortic rings. Its vasodilator properties were tested in coronary arteries using Langendorff‐perfused rat hearts. Finally, erucin's antihypertensive activity was evaluated in vivo in normotensive and spontaneously hypertensive rats (SHRs) by recording systolic BP using the tail‐cuff method.Key ResultsErucin induced the release of H2S inside HASMCs. Moreover, erucin hyperpolarized the membrane of HASMCs membrane in a concentration‐dependent manner. It induced vasodilatation of rat aortic rings, in endothelium‐denuded vessels. This effect was further improved by the presence of endothelial NO. When pre‐incubated with rat aortic rings, erucin induced concentration‐dependent inhibition of noradrenaline‐induced vasoconstriction. Erucin did not affect basal coronary flow but restored the flow to normal in pre‐contracted coronary vessels. Finally, in vivo, erucin decreased systolic BP in SHRs by about 25%, and restored the BP to values observed in normotensive rats.Conclusions and ImplicationsErucin is an H2S donor endowed with vasorelaxing and antihypertensive effects.Linked ArticlesThis article is part of a themed section on Hydrogen Sulfide in Biology & Medicine. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v177.4/issuetoc