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Methyl jasmonate (MeJA) is a plant growth regulator, which plays an important role in signaling defense response and regulating plant secondary metabolism. Here, broccoli hairy roots growing for 18 d were treated with MeJA for 0, 3, 6, 9, and 12 h, respectively. Compared with the 0 h control group, the content of glucoraphanin (GRA) and sulforaphane (SF) was increased under MeJA treatment for 3, 6, 9, and 12 h. The activity of myrosinase (MYR) was reduced in broccoli hairy roots. With the broccoli (Brassica oleracea L. var. botrytis L) genome as the reference genome, 4733 DEGs (differentially expressed genes) were detected in the hairy roots that were treated by MeJA for 0, 3, 6, 9, and 12 h; among them, 1024 genes were upregulated and 3709 genes were downregulated. The pathway of “SNARE interaction in vesicular transport” (ko04130) was ranked 1st by KEGG pathway enrichment analysis of commonly upregulated differential genes at 0, 3, 6, 9, and 12 h. The expression of four key genes ABCB19, ABCG6, ABCG36, and ABCB9, which regulated the release of GRA and SF into a liquid medium, was upregulated at 0, 3, 6, and 9 h, respectively. It revealed that the release of GRA and SF from hairy roots was related to the ABC transporter gene family and SNARE protein-mediated exocytosis by transcriptome analysis.

Insects affect crop harvest yield and quality, making plant response mechanisms to insect herbivores a heavily studied topic. However, analysis of plant responses to honeybees is rare. In this study, comprehensive metabolic profiling of Arabidopsis thaliana exposed to honeybees was performed to investigate which metabolites were changed by the insect. A total of 85 metabolites―including chlorophylls, carotenoids, glucosinolates, policosanols, tocopherols, phytosterols, β-amyrin, amino acids, organic acids, sugars, and starch―were identified using high performance liquid chromatography, gas chromatography-mass spectrometry, and gas chromatography-time-of-flight mass spectrometry. The metabolite profiling analysis of Arabidopsis exposed to honeybees showed higher levels of stress-related metabolites. The levels of glucosinolates (glucoraphanin, 4-methoxyglucobrassicin), policosanols (eicosanol, docosanol, tricosanol, tetracosanol), tocopherols (β-tocopherol, γ-tocopherol), putrescine, lysine, and sugars (arabinose, fructose, glucose, mannitol, mannose, raffinose) in Arabidopsis exposed to honeybees were higher than those in unexposed Arabidopsis. Glucosinolates act as defensive compounds against herbivores; policosanols are components of plant waxes; tocopherols act as an antioxidant; and putrescine, lysine, and sugars contribute to stress regulation. Our results suggest that Arabidopsis perceives honeybees as a stress and changes its metabolites to overcome the stress. This is the first step to determining how Arabidopsis reacts to exposure to honeybees.

Sulforaphene (SRP) is the main isothiocyanate produced by myrosinase hydrolysis of glucoraphanin from Cruciferae plants. This review discusses the latest results on the formation, content, and stab...

Discoveries in model plants grown under optimal conditions can provide important directions for crop improvement. However, it is important to verify whether results can be translated to crop plants grown in the field. In this study, we sought to study the role of MYB28 in the regulation of aliphatic glucosinolate (A-GSL) biosynthesis and associated sulfur metabolism in field-grown Brassica oleracea with the use of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 gene-editing technology. We describe the first myb28 knockout mutant in B. oleracea, and the first CRISPR field trial in the United Kingdom approved and regulated by the UK Department for Environment, Food & Rural Affairs after the reclassification of gene-edited crops as genetically modified organisms by the European Court of Justice on July 25, 2018. We report that knocking out myb28 results in downregulation of A-GSL biosynthesis genes and reduction in accumulation of the methionine-derived glucosinolate, glucoraphanin, in leaves and florets of field-grown myb28 mutant broccoli plants, whereas accumulation of sulfate, S-methyl cysteine sulfoxide, and indole glucosinolate in leaf and floret tissues remained unchanged. These results demonstrate the potential of gene-editing approaches to translate discoveries in fundamental biological processes for improved crop performance.

Brassica sprouts are known as a good source of antimicrobial bioactive compounds such as phenolics and glucosinolates (GLs). We aim at understanding how He-Ne laser light treatment (632 nm, 5 mW) improves sprout growth and physiology and stimulates the accumulation of bioactive metabolites in three Brassica spp., i.e., mustard, cauliflower, and turnip. Moreover, how these changes consequently promote their biological activities. Laser light improved growth, photosynthesis, and respiration, which induced the accumulation of primary and secondary metabolites. Laser light boosted the levels of pigments, phenolics, and indole and aromatic precursors of GLs, which resulted in increased total GLs and glucoraphanin contents. Moreover, laser light induced the myrosinase activity to provoke GLs hydrolysis to bioactive sulforaphane. Interestingly, laser light also reduced the anti-nutrient content and enhanced the overall biological activities of treated sprouts including antioxidant, antibacterial, anti-inflammatory, and anticancer activities. Accordingly, laser light is a promising approach for boosting the accumulation of beneficial metabolites in Brassica sprouts and, subsequently, their biological activities.

Isothiocyanates (ITCs) are well-known chemopreventive agents that have received significant interest across the nutrition and pharmaceutical industries owing to their anticancer properties, thus it is essential to increase the conversion of glucosinolate to ITCs by myrosinase to maximize their health benefits. In this paper, using broccoli seed meals as a raw material, we comparatively analyzed the outcomes of two extraction methods: (i) hydrolysis followed by extraction (HFE) and (ii) simultaneous hydrolysis and extraction (SHE) in terms of the ITC yield. The results revealed that the SHE method showed a relatively greater erucin production from broccoli seeds and greater antitumor and antioxidant activities. A similar phenomenon was found for the hydrolysates of crude myrosinase and crude glucosinolate separated from broccoli seeds. However, when the crude glucosinolates were hydrolyzed by purified broccoli myrosinase, or when pure glucoraphanin was hydrolyzed by crude myrosinase, no significant effects were noted on the types and yields of ITCs between the SHE and HFE methods.

Exogenous melatonin (10 μM) enhances ZnSO4 (4 mM) stress tolerance and regulates the isothiocyanate content of broccoli sprouts. Nevertheless, the molecular mechanism underlying the role of melatonin in isothiocyanate metabolism under ZnSO4 stress is unclear. The effects of exogenous melatonin on growth and isothiocyanate metabolism in broccoli sprouts under ZnSO4 stress during germination were investigated by physio-biochemical methods, quantification of relative gene expression levels, and the isobaric tags for the relative and absolute quantitation (iTRAQ) labelling technique. Compared with sprouts under ZnSO4 stress alone, sprout length, fresh weight and free calcium content increased significantly in sprouts under ZnSO4 stress plus melatonin treatment while electrolyte leakage and malonaldehyde content decreased. The glucosinolate content and myrosinase activity also significantly increased in sprouts under ZnSO4 stress plus melatonin treatment compared with the control, and thus the isothiocyanate and sulforaphane content increased markedly. Meanwhile, the expression of glucoraphanin biosynthesis genes, such as MYB28, CYP83A1, AOP2, BoSAT1, and BoHMT1 was significantly induced by melatonin in sprouts under ZnSO4 stress. Furthermore, compared with sprouts under ZnSO4 stress alone, a total of 145 proteins in broccoli sprouts under ZnSO4 stress plus melatonin treatment showed differential relative abundances. These proteins were divided into 13 functional classes and revealed that pathways for sulfur metabolism, glucosinolate biosynthesis, selenocompound metabolism, biosynthesis of secondary metabolites and peroxisome were significantly enriched. The present study indicates that exogenous melatonin alleviates the adverse effects of ZnSO4 stress on sprout growth and promotes glucoraphanin biosynthesis and the hydrolysis of glucoraphanin to form isothiocyanates in broccoli sprouts.

Purpose Studies on broccoli (Brassica oleracea var. italica) indicate beneficial effects against a range of chronic diseases, commonly attributed to their bioactive phytochemicals. Sulforaphane, the bioactive form of glucoraphanin, is formed by the action of the indigenous enzyme myrosinase. This study explored the role that digestion and cooking practices play in bioactivity and bioavailability, especially the rarely considered dose delivered to the colon. Methods The antimicrobial activity of sulforaphane extracts from raw, cooked broccoli and cooked broccoli plus mustard seeds (as a source myrosinase) was assessed. The persistence of broccoli phytochemicals in the upper gastrointestinal tract was analysed in the ileal fluid of 11 ileostomates fed, in a cross-over design, broccoli soup prepared with and without mustard seeds. Results The raw broccoli had no antimicrobial activity, except against Bacillus cereus, but cooked broccoli (with and without mustard seeds) showed considerable antimicrobial activity against various tested pathogens. The recovery of sulforaphane in ileal fluids post soup consumption was Escherichia coli K12, no inhibitory effects were observed. Analysis of glucosinolates composition in ileal fluids revealed noticeable inter-individual differences, with six “responding” participants showing increases in glucosinolates after broccoli soup consumption. Conclusions Sulforaphane-rich broccoli extracts caused potent antimicrobial effects in vitro, and the consumption of sulforaphane-enriched broccoli soup may inhibit bacterial growth in the stomach and upper small intestine, but not in the terminal ileum or the colon.

Aging is caused by an imbalance between antioxidants and ROS. Nuclear Factor Erythroid 2-related factor 2 (Nrf2) is a transcription factor that regulates antioxidant genes. Under normal conditions, Nrf2 will bind keap1 and cause degradation of Nrf2. Nrf2 activation can be stimulated by secondary metabolites, such as glucosinolate (glucoraphanin and sulforaphane) and phenolic (kaempferol and quercetin) groups found in broccoli ( Brassica oleracea ). The purposes of this study were to analyze the interaction of the four compounds with Keap1 through molecular docking, to identify interactions that inhibit Keap1, and also to know the bioactivity scores, drug-likeness, and bioactivity prediction of each compound. The Nrf2-Keap1 protein (ID: 2FLU) structure was retrieved from the protein database, whereas the quercetin (CID: 5280343); kaempferol (CID: 5280863), sulforaphane (CID: 5350), and glucoraphanin (CID: 656556) were obtained from the PubChem Database. Molecular docking was done with HEX 8.0. The docking results were visualized with Discovery Studio 2020. Drug-likeness and bioactivity scores of the compounds were identified using mollinspiration. Prediction of bioactivity was carried out with PASS Online. The results showed that the binding energy of quercetin with Keap1 was -268.72 kcal.mol -1 , and glucoraphanin with Keap1 was -318.01 kcal.mol -1 . We found that quercetin from the phenolic group and glucoraphanin from the glucosinolate group had a strong interaction with Keap1, indicated by the number of interactions occurred and the smaller energy needed. Hence both compounds could inhibit the interaction of Keap1-Nrf2. Consequently, Nrf2 could transcribe antioxidant genes. The interaction between Keap1 and quercetin may play a role related to ROS reduction activities, such as enhancing HMOXI expression. This study indicates that quercetin has more potential in drug development as peroxidase inhibitors. Keyword: Aging, bioinformatic, glucoraphanin, keap1, quercetin

Aim Epidemiological data suggest that maternal immune activation (MIA) plays a role in the etiology of neuropsychiatric disorders including autism spectrum disorder (ASD) and schizophrenia. However, there is no prophylactic nutrition that can prevent the onset of neurodevelopmental disorders in offspring after MIA. The aim of this study was undertaken to examine whether dietary intake of glucoraphanin (GF: the precursor of a natural anti‐inflammatory compound sulforaphane) can prevent the onset of behavioral abnormalities in offspring after MIA. Methods One percent of GF food pellet or normal food pellet was given into female mice during pregnancy and lactation (from E5 to P21). Saline (5 mL/kg/d) or poly(I:C) (5 mg/kg/d) was injected into pregnant mice from E12 to E17. Behavioral tests and immunohistochemistry of parvalbumin (PV) were performed in male offspring. Results Dietary intake of GF during pregnancy and lactation prevented cognitive deficits and social interaction deficits in the juvenile offspring after MIA. Furthermore, dietary intake of GF during pregnancy and lactation prevented cognitive deficits in the adult offspring after MIA. Moreover, dietary intake of GF prevented the reduction of PV immunoreactivity in the medial prefrontal cortex of adult offspring after MIA. Conclusion These data suggest that dietary intake of GF during pregnancy and lactation could prevent behavioral abnormalities in offspring after MIA.

In this study, we found that the glucoraphanin (GR) content in broccoli seedlings decreased before first foliage leaf emergence. Methyl jasmonate (MeJA) treatment significantly increased the GR content in broccoli, with effects being consistent over the period of application. Unlike MeJA, abscisic acid (ABA) and salicylic acid (SA) led to slight GR accumulation within 24 h of the application. The feeding of a sulfur-containing compound, MgSO4, via the root of broccoli caused accumulation of GR within 24 h. Thus, both the germination time and the duration of sulfur fertilization and MeJA application are important to maximize GR content in broccoli.

Evolution of important glucosinolates (GLSs), namely, sinigrin, glucoraphanin, glucoerucin and glucobrassicin, in three commonly consumed Brassica vegetables viz. white cabbage, Chinese cabbage and bok choy during their processing into vegetable powder was investigated. Drying was noted to be a major processing step causing significant losses of GLSs. Interestingly, different GLSs and even the same GLSs in different vegetables showed different thermal stabilities during drying. The stability of GLSs in vegetable powder during in vitro gastric digestion was also studied. Glucoraphanin exhibited the highest stability while glucobrassicin was the most vulnerable GLS under in vitro gastric conditions. White cabbage is found to be a promising material for the production of vegetable powder as it contains high contents of GLSs, especially glucoraphanin and glucoerucin, which are important precursors of anticarcinogenic compounds, namely sulforaphane and erucin. These two GLSs were also noted to be stable during in vitro gastric digestion.

To date studies revealed that MYB28 intensively involved biosynthesis of GSLs, there is no direct evidence of MYB28 in glucoraphanin (GR) biosynthesis of broccoli. Furthermore, Agrobacterium (Agro)-infiltration also induced GSLs in Arabidopsis. However, there is no information regarding Agro-dependent GR accumulation in broccoli. We revealed that over-expression of BroMYB28 could accumulate GR, and Agro-infiltration is also enough to induce the accumulation of GR in broccoli. From our results, it is suggested that they used different regulatory mechanisms; BroMYB28 over-expression induced both BroMYB28 and BroMYB29 (Bol008849-like) transcripts; Agro-infiltration induced only BroMYB29 (Bol008849-like) transcripts.

The effects of S and Se treatment on cabbage, especially the interactions of S and Se metabolism with the biosynthesis of glucosinolate (GSL), including glucoraphanin, which is a major aliphatic GSL in cruciferous vegetables and the precursor of the anticancer compound sulforaphane, were examined. Cabbage plants were treated with sulfate and selenite (SeO32−), and the total S, Se, and GSL contents of cabbage head and outer foliage leaves were measured. Results showed that selenite treatment was beneficial to GSL biosynthesis and Se accumulation in cabbage head and outer foliage leaves. GSL synthesis was induced by exogenous selenite-elevated sulfate treatment at certain concentration ratios, i.e., 50-μΜ selenite + 1-mΜ sulfate or 100-μΜ selenite + 4-mΜ sulfate. A high exogenous sulfate concentration was more favorable to GSL accumulation than a low sulfate concentration. According to the relative expression of genes on GSL synthesis, an increase in the GSL content was attributed to the upregulation of gene expression and possible transportation from the outer foliage leaf to the head of cabbage. These results might be helpful for increasing the health benefits of cabbage by supplying exogenous S and Se. Further research should explore the effects of sulfate and selenite on GSL precursor substances to reveal the reason why total GSL contents increased.

Chinese kale (Brassica oleracea var. alboglabra) has high nutritional value. This study investigated the contents of glucosinolates, antioxidants (chlorophylls, carotenoids, vitamin C, and total phenolics), and antioxidant capacity in five organs from six varieties of Chinese kale. The highest concentrations of individual and total glucosinolates were in the roots and inflorescences, respectively. The highest levels of antioxidants and antioxidant capacity were in inflorescences and leaves. Plant organs played a predominant role in glucosinolate and antioxidant accumulation. Glucoiberin, glucoraphanin, and glucobrassicin, the main anticarcinogenic glucosinolates, could be enhanced simultaneously because of their high positive correlations. The relationship between glucosinolates and antioxidant capacity indicated that glucobrassicin might contribute to the total antioxidant capacity. These results provide useful information related to consumption, breeding of functional varieties, and use of the non-edible organs of Chinese kale.

Glucosinolates are secondary plant metabolites present in Brassica vegetables. The endogenous enzyme myrosinase is responsible for the hydrolysis of glucosinolates, yielding a variety of compounds, including health-promoting isothiocyanates. The influence of cabbage accession and growing conditions on myrosinase activity, glucosinolates (GSL) and their hydrolysis products (GHPs) of 18 gene-bank cabbage accessions was studied. Growing conditions, cabbage morphotype and accession all significantly affected myrosinase activity and concentration of glucosinolates and their hydrolysis products. In general, cabbages grown in the field with lower growth temperatures had significantly higher myrosinase activity than glasshouse samples. Profile and concentration of glucosinolates and their hydrolysis products differed across the accessions studied. Aliphatic glucosinolates accounted for more than 60 % of total glucosinolates in most of the samples assessed. Nitriles and epithionitriles were the most abundant hydrolysis products formed. The results obtained showed that consumption of raw cabbages might reduce the amount of beneficial hydrolysis products available to the consumer, as more nitriles were produced from hydrolysis compared to beneficial isothiocyanates. However, red and white cabbages contained high concentrations of glucoraphanin and its isothiocyanate, sulforaphane. This implies that careful selection of accessions with ample concentrations of certain glucosinolates can improve the health benefits derived from raw cabbage consumption.

Broccoli sprouts are rich in health-promoting bioactive compounds. Their content depends on both cultivation light quality and temperature. However, these effects have been previously addressed in isolation. Here, the dual inputs of cultivation light quality [blue (B), red (R), mixture of R and B (R+B), mixture of R and UVA (R+UVA)] and air temperature (15, 19, and 23 °C) on determining growth, external quality, and the cotyledon and hypocotyl content of five major bioactive compounds were investigated. The carbohydrate status at harvest and postharvest ratio of variable to maximum fluorescence (Fv/Fm) were also assessed. Hypocotyl length was generally enhanced under monochromatic light (R or B) and elevated temperature. Total phenolic, total flavonoid, and glucoraphanin contents were generally higher in cotyledon as compared to hypocotyl. Hypocotyl anthocyanin, total phenolic, total flavonoid, and ascorbic acid contents were generally enhanced by R+B, and were decreased by R. Cotyledon content in these metabolites was generally stimulated by B, and reduced under R or R+UVA. Temperature affected metabolite content depending on the metabolite, organ, and light quality. Lower temperatures, R (23 °C) or R+UVA (15, 19, and 23 °C) were associated with decreased postharvest Fv/Fm. In conclusion, low cultivation temperature (&lt, 23 °C), as well as R or R+UVA ought to be avoided. Instead, B and R+B are suitable, with B being preferable, owing to better external quality and enhanced metabolite content in cotyledon which generally holds higher content than hypocotyl.

Hydrolysis of glucobrassicin by plant or bacterial myrosinase produces multiple indoles predominantly indole-3-carbinol (I3C). I3C and its majorin vivoproduct, 3,3'-diindolylmethane (DIM), are effective cancer chemopreventive agents in pre-clinical models and show promise in clinical trials. The pharmacokinetics/pharmacodynamics of DIM have been studied in both rodents and humans and urinary DIM is a proposed biomarker of dietary intake of cruciferous vegetables. Recent clinical studies at Oregon State University show surprisingly robust metabolism of DIMin vivowith mono- and di-hydroxylation followed by conjugation with sulfate or glucuronic acid. DIM has multiple mechanisms of action, the most well-characterized is modulation of aryl hydrocarbon receptor (AHR) signaling. In rainbow trout dose-dependent cancer chemoprevention by dietary I3C is achieved when given prior to or concurrent with aflatoxin B1, polycyclic aromatic hydrocarbons, nitrosamines or direct acting carcinogens such as N-methyl-N'-nitro-nitrosoguanidine. Feeding pregnant mice I3C inhibits transplacental carcinogenesis. In humans much of the focus has been on chemoprevention of breast and prostate cancer. Alteration of cytochrome P450-dependent estrogen metabolism is hypothesized to be an important driver of DIM-dependent breast cancer prevention. The few studies done to date comparing glucobrassicin-rich crucifers such as Brussels sprouts with I3C/DIM supplements have shown the greater impact of the latter is due to dose. Daily ingestion of kg quantities of Brussels sprouts is required to producein vivolevels of DIM achievable by supplementation. In clinical trials these supplement doses have elicited few if any adverse effects. Sulforaphane from glucoraphanin can act synergistically with glucobrassicin-derived DIM and this may lead to opportunities for combinatorial approaches (supplement and food-based) in the clinic.

Glucoraphanin (GRA) is found in the seeds and vegetative organs of broccoli (Brassica oleracea L. var. italica Planch) as the precursor of anti-carcinogen sulforaphane (SF). The yield of GRA obtained from these materials is weak and the cost is high. In recent years, the production of plant secondary metabolites by large-scale hairy roots culture in vitro has succeeded in some species. Melatonin (MT) is a natural hormone which existed in numerous organisms. Studies have demonstrated that MT can improve the synthesis of secondary metabolites in plants. At present, it has not been reported that MT regulates the biosynthesis of glucoraphanin in broccoli hairy roots. In this study, the broccoli hairy roots that grew for 20 d were respectively treated by 500 µM MT for 0, 6, 12, 20 and 32. To explore the reason of changes in secondary metabolites and reveal the biosynthetic pathway of glucoraphanin at transcriptional level. Compared with 0 h, the yield of GRA under other treatments was increased, and the overall trend was firstly increased and then decreased. The total yield of GRA reached the highest at 12 h, which was 1.22-fold of 0 h. Then, the genome of broccoli as the reference, a total of 13234 differentially expressed genes (DEGs) were identified in broccoli hairy roots under treatment with 500 µM MT for 0, 6, 12, 20 and 32 h, respectively. Among these DEGs, 6266 (47.35%) were upregulated and 6968 (52.65%) were downregulated. It was found that the pathway of ‘Glucosinolates biosynthesis (ko00966)’ was enriched in the 16th place by Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis of the upregulated DEGs. The expression of key genes in the GRA biosynthesis pathway was upregulated at all time points, and a deduced GRA biosynthesis pathway map was constructed for reference.

Red cabbage (Brassica oleracea L. var. capitata) continues to receive increasing attention on its health-promoting properties because of its high glucosinolate content. Glucosinolates are an unstable active substance, however, there are few studies on their changes in different cooking processes. In this study, we investigated the effects of processing methods (boiling, steaming, microwave heating, frying, stir-frying) and boiling time on glucosinolates in red cabbage. Ten glucosinolates, including 4-methoxyglucobrassicin, neoglucobrassicin, glucoalyssin, glucobrassicin, glucoraphanin, glucoiberin, progoitrin, gluconapin and sinigrin, in red cabbage were detected. Decreases of 32.36%, 24.83%, 25.27%, 81.11% and 84.29% for total glucosinolates were observed after boiling, microwaving, steaming, frying and stir-frying. Indole glucosinolates were more efficiently lost compared to aliphatic glucosinolates after boiling, while microwaving, steaming, frying and stir-frying also resulted in a greater reduction in indole glucosinolates than aliphatic glucosinolates. Glucoalyssin, glucoerucin and sinigrin were more thermal sensitive than other glucosinolates. It was confirmed that microwaving and steaming retained higher levels of glucosinolates than other methods and may be better for cooking red cabbage.

Addition of selenium or application of ultraviolet A (UVA) radiation for crop production could be an effective way of producing phytochemical-rich food. This study was conducted to investigate the effects of selenium and UVA radiation, as well as their combination on growth and phytochemical contents in broccoli microgreens. There were three treatments: Se (100 μmol/L Na2SeO3), UVA (40 μmol/m2/s) and Se + UVA (with application of Se and UVA). The control (CK) was Se spraying-free and UVA radiation-free. Although treatment with Se or/and UVA inhibited plant growth of broccoli microgreens, results showed that phytochemical contents increased. Broccoli microgreens under the Se treatment had higher contents of total soluble sugars, total phenolic compounds, total flavonoids, ascorbic acid, Fe, and organic Se and had lower Zn content. The UVA treatment increased the contents of total chlorophylls, total soluble proteins, total phenolic compounds, and FRAP. However, the Se + UVA treatment displayed the most remarkable effect on the contents of total anthocyanins, glucoraphanin, total aliphatic glucosinolates, and total glucosinolates, here, significant interactions between Se and UVA were observed. This study provides valuable insights into the combinational selenium and UVA for improving the phytochemicals of microgreens grown in an artificial lighting plant factory.

Broccoli microgreens have shown potential health benefits due to their high glucosinolate (GL) levels. Previously, we observed that postharvest UVB treatment did not have much effect on increasing GLs in broccoli microgreens. In this study, we investigated the influence of preharvest UVB irradiation on GL levels in broccoli microgreens. UHPLC-ESI/ITMS analysis showed that preharvest UVB treatments with UVB 0.09 and 0.27 Wh/m2 significantly increased the glucoraphanin (GLR), glucoerucin (GLE), and total aliphatic GL levels by 13.7 and 16.9%, respectively, in broccoli microgreens when measured on harvest day. The nutritional qualities of UVB-treated microgreens were stable during 21-day storage, with only small changes in their GL levels. Broccoli microgreens treated before harvest with UVB 0.27 Wh/m2 and 10 mM CaCl2 spray maintained their overall quality, and had the lowest tissue electrolyte leakage and off-odor values during the storage. Furthermore, preharvest UVB 0.27 Wh/m2 treatment significantly increased GL biosynthesis genes when evaluated before harvest, and reduced the expression level of myrosinase, a gene responsible for GL breakdown during postharvest storage. Overall, preharvest UVB treatment, together with calcium chloride spray, can increase and maintain health-beneficial compound levels such as GLs and prolong the postharvest quality of broccoli microgreens.

Seeds of 32 pure lines and 6 commercial broccoli cultivars were used to investigate variation in glucosinolates and their breakdown products. The aliphatic glucosinolate content was 54.5-218.7 μmol/g fresh weight, accounting for >90% of the total glucosinolates. The major glucosinolates found were glucoraphanin and glucoerucin in 27 samples and progoitrin in 7 samples. A gas chromatography-flame ionization detector (GC-FID) method was used to identify glucosinolate breakdown products; nine products were directly determined using standards. Using Arabidopsis thaliana lines myb28myb29 and Landsberg erecta to hydrolyze each reference glucosinolate, seven products were tentatively identified. 4-(Methylsulfinyl)butyl isothiocyanate and 5-(methylsulfinyl)pentanenitrile contents were 2.6-91.1 μmol/g fresh weight and 0-35.4 μmol/g fresh weight, respectively, with epithionitriles being more common than nitriles in accessions rich in alkenyl glucosinolate. Additionally, (S)-5-vinyl-1,3-oxazolidine-2-thione was detected in accessions rich in progoitrin. Specific lines with altered glucosinolate profiles and breakdown products were obtained and discussed according to the putative glucosinolate metabolism pathway.

In this study, the glucosinolates (GLs) content and isothiocyanates (ITCs) formation in broccoli sprouts and their dynamic variation during hydrolysis were investigated. The results showed that 7 kinds of GLs were detected and glucoraphanin (GRA) was the dominating component. After hydrolysis, three kinds of ITCs and 2 kinds of nitriles were detected in broccoli sprouts, however, the corresponding nitriles of 4-isothiocyanato-1-butene and 1-isothiocyanato-butane were not detectable. This inferred that the inter-transformation among ITCs existed during sprouts hydrolysis. According to the identification, 4-isothiocyanato-1-butene had another source in addition to gluconapin (GNA), 1-isothiocyanato-butane stemmed from glucoerucin (GER). Moreover, the affinity of MYR in broccoli sprouts was stronger to GER than to GRA and other GLs.

The aim of the study was to determine the effect of pretreatment with hot water or steaming on glucosinolates, polyphenols contents and antioxidant capacity in obtained raw juices. Moreover, in vitro cytotoxic activity of the raw juice to the cells derived from the gastrointestinal tract, including the small intestine (IEC-6 cell line), colon (Caco-2 cell line) and the liver (HepG2 cell line) were also investigated. The dominant glucosinolates in the wild rocket leaves were glucoraphanin (36%) and dimeric 4-mercaptobutyl (30%), followed by glucosativin and glucoerucin, 11% per each. Glucothiobeinin (6%), glucobrassicin (1%), 4-methoxyglucobrassicin (1%) and two unidentified compounds (4%) were also detected in rocket leaves. In terms of phenolic compounds, quercetin constituted the majority (55%) and the rest composed of hydroxycinnamic acids. In raw juices produced from steamed, pretreatment with hot water and untreated (control) leaves, glucosinolate contents were lower about 21%, 37% and 53%, respectively, than their levels in the raw material. The highest content of polyphenols among the juices tested (45.4 mg/100 g fresh weight) and antioxidant capacity (5.8 µmol Trolox/1 g f.w.) was recorded in the raw juice from pretreated leaves with hot water. The wild rocket raw juice concentrations responsible for a 50% reduction in Caco-2 and HepG2 cell viability were estimated at 1.87 ± 0.08 mg/mL and 3.54 ± 0.29 mg/mL. The viability of the IEC-6 cells was reduced by only 19.04%, at the maximum concentration (3.6 mg/mL) of the raw juice.

This study investigated the effect of lactic acid bacteria (LAB) fermentation on the chemical profile of autoclaved broccoli puree, using 7 broccoli-derived LAB isolates (named F1-F5, BF1 and BF2). The total concentrations of glucosinolates (glucoiberin, progoitrin and glucoraphanin) and 10 major phenolics significantly increased from trace level and 289 μg total phenolics/g dry weight (DW) respectively in autoclaved broccoli to 55 to ∼359 μg/g DW and 903 to ∼3105 μg/g DW respectively in LAB fermented broccoli puree. Differential impacts of LAB isolates on the chemical composition of autoclaved broccoli were observed, with the major differences being the significant increase in phloretic acid after fermentation by F1-F5 and an elevated glucoraphanin level in ferments by F1 and BF2. LAB fermentation is a promising way to increase the content of glucosinolates and polyphenolic compounds in broccoli, making the ferments attractive for use as functional ingredients or as a whole functional food.

In this study, we determined the contents of glucosinolate, polyphenol, and flavonoid, and the antioxidant activities of uncooked, steamed, and boiled cauliflower. Eight glucosinolate peaks were detected, representing glucoiberin, progoitrin, glucoraphanin, sinigrin, gluconapin, glucoiberverin, glucobrassicin, and gluconasturtiin. Boiled cauliflower contained significantly lowered concentrations of glucosinolate, total polyphenol, and total flavonoid compared to uncooked or steamed cauliflower. These results clearly indicate that health-promoting compounds in cauliflower are significantly impacted by different cooking methods: uncooked> steamed> boiled. The amounts of total polyphenols and total flavonoids in uncooked cauliflower extracted with 80% ethanol were higher than extracts of steamed and boiled cauliflower. The highest antioxidant activity was observed in uncooked cauliflower extracted using 80% ethanol compared to those extracted with water at the same concentration. Steamed and boiled cauliflower extracts also showed lower antioxidant activity than uncooked extracts. Based on these results, fresh uncooked cauliflower contains higher contents of health-promoting compounds and elevated antioxidant activity. Moreover, steaming may be more desirable than boiling in order to minimize loss of glucosinolates when storing, pretreating, processing, and cooking cruciferous vegetables.

Aims Parkinson's disease (PD) is a chronic and progressive neurodegenerative disorder. Although diet may influence the development of PD, the precise mechanisms underlying relationship between diet and PD pathology are unknown. Here, we examined whether dietary intake of glucoraphanin (GF), the precursor of a natural antioxidant sulforaphane in cruciferous vegetables, can affect the reduction of dopamine transporter (DAT) in the mouse striatum after repeated administration of MPTP (1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine). Methods Normal food pellet or 0.1% GF food pellet was given into male mice for 28 days from 8‐week‐old. Subsequently, saline (5 mL/kg × 3, 2‐hour interval) or MPTP (10 mg/kg × 3, 2‐hour interval) was injected into mice. Immunohistochemistry of DAT in the striatum was performed 7 days after MPTP injection. Results Repeated injections of MPTP significantly decreased the density of DAT‐immunoreactivity in the mouse striatum. In contrast, dietary intake of 0.1% GF food pellet significantly protected against MPTP‐induced reduction of DAT‐immunoreactivity in the striatum. Conclusion This study suggests that dietary intake of GF food pellet could prevent MPTP‐induced dopaminergic neurotoxicity in the striatum of adult mice. Therefore, dietary intake of GF‐rich cruciferous vegetables may have beneficial effects on prevention for development of PD., Oxidative stress plays a role in the neurotoxicity of MPTP in the striatum. Dietary intake of glucoraphanin could prevent the MPTP‐induced neurotoxicity in mouse striatum. Dietary intake of glucoraphanin‐rich vegetables may have beneficial effects on prevention of Parkinson's disease.

A new cyclodextrin-mediated capillary zone electrophoresis method for the separation and quantification of intact glucosinolates (GSL) has been developed. β-Cyclodextrin concentration and temperature were shown to be key parameters for selectivity. Controlled variation of both parameters provided a set of methods for the analysis of GSL-containing plants with customized selectivities. As a proof of concept, parameters were adapted for the analysis of broccoli-based dietary supplements (BBDS) obtained from different sources. All main GSL (glucoraphanin, -iberin, -erucin, and 4-hydroxyglucobrassicin) – which typically occur in broccoli plants at early development stages – were successfully analyzed using a 100 mM borate buffer with 1% β-cyclodextrin at 20 °C. Analysis of BBDS showed unexpected results. In most cases, supplements had been specified to contain seeds, sprouts or young broccoli plants. In many cases, results did not reflect the description on the label. Several samples apparently contained broccoli from older plant development stages, marked by higher amounts of indole-derived GSL and very low concentrations of aliphatic GSL. Considerable amounts of non-broccoli GSL were detected in two samples: sinalbin and a mixture of 3-butenyl- and 4-pentenylglucosinolate. No glucosinolates were detectable in two further samples. In total >50% of all investigated samples did not meet the specifications.

Background Epidemiological evidence suggests that consumption of cruciferous vegetables is associated with reduced risk of prostate cancer progression, largely attributed to the biological activity of glucosinolate degradation products, such as sulforaphane derived from glucoraphanin. Because there are few therapeutic interventions for men on active surveillance for prostate cancer to reduce the risk of cancer progression, dietary approaches are an appealing option for patients. Objective We evaluated whether consumption of a glucoraphanin-rich broccoli soup for 1 y leads to changes in gene expression in prostate tissue of men with localized prostate cancer. Methods Forty-nine men on active surveillance completed a 3-arm parallel randomized double-blinded intervention study for 12 mo and underwent transperineal template biopsy procedures and dietary assessment at the start and end of the study. Patients received a weekly 300 mL portion of soup made from a standard broccoli (control) or from 1 of 2 experimental broccoli genotypes with enhanced concentrations of glucoraphanin, delivering 3 and 7 times that of the control, respectively. Gene expression in tissues from each patient obtained before and after the dietary intervention was quantified by RNA sequencing followed by gene set enrichment analyses. Results In the control arm, there were several hundred changes in gene expression in nonneoplastic tissue during the 12 mo. These were associated with an increase in expression of potentially oncogenic pathways including inflammation processes and epithelial–mesenchymal transition. Changes in gene expression and associated oncogenic pathways were attenuated in men on the glucoraphanin-rich broccoli soup in a dose-dependent manner. Although the study was not powered to assess clinical progression, an inverse association between consumption of cruciferous vegetables and cancer progression was observed. Conclusion Consuming glucoraphanin-rich broccoli soup affected gene expression in the prostate of men on active surveillance, consistent with a reduction in the risk of cancer progression. This trial was registered at clinicaltrials.gov as NCT01950143.

This study aimed to investigate the glyoxal (GO) scavenging effect of broccoli and wine sulphur compounds in model systems under physiological conditions. The model systems composed of GO and different sulphur compounds both in pure forms and food extracts were subjected to simulated physiological conditions (37 °C & 7.4 pH, 2 h). The results revealed that some of these sulphur sources could scavenging GO under these conditions. From the pure sulphur compounds tested, potassium metabisulphite and cysteine was found as the most effective. Wine scavenged 18.01 ± 2.83% of GO (10 µmole) whereas potassium metabisulphite addition (7 µmole) to wine caused additional 20.81% scavenge. In addition, the results indicated that 11.70 ± 7.07% and 54.48 ± 6.38% of GO was scavenged by bioaccessible raw and steamed broccoli, respectively. High-resolution mass spectrometry scan analysis confirmed that it was attributed to the addition of GO to both amino acids and sulforaphane (SF)-amino acid adducts under physiological conditions.

Background: Sulforaphane (SF, 1-isothiocyanato-4-(methyl-sulfinyl)-butane) is found in broccoli, cabbage and cauliflower. Methods: we performed a critical review on the antioxidative, chemopreventive and antitumor effects of SF from cruciferous vegetables against prostate cancers and molecular pathways. For a complete and reliable review, primary and secondary resources were used, including original and review articles, books and government documents published until March 2018. Articles that are in duplicity and disconnected are not considered for review. SF is derived from glucoraphanin (4-methyl-sulfinyl-butyl-glucosinate), being one of the most commonly found isothiocyanates in vegetables from Brassica spp., especially in broccoli samples. In vitro studies indicate that SF induces apoptosis in a dependent or non-dependent method of androgens by transcription of tumor suppressor genes, oxidation response and higher expression of phase II enzymes in prostate cancer cells. Sulforaphane also decreases transcription of the nuclear factor kB and antiapoptotic proteins, expression of cyclin D2 and survivin and DNA synthesis, increases Nrf2 gene activity, interferes with genome compacting by inhibition of histone deacetylases and disrupts Hsp90 complexes, which cause cell cycle arrest, mitosis interruption, activation of caspases and mitochondria depolarization. Conclusion: SF and cruciferous vegetables play antioxidative and chemopreventive role, delaying or blocking in vivo carcinogenesis, causing biochemical and epigenetic changes, preventing, delaying, or reversing preneoplastic or advanced prostate lesions, and frequently activating tumor cell death by intrinsic methods of apoptosis. These outcomes encourage the consumption of Brassica specimens, which could be easily achieved by the incorporation of food and vegetables rich in cruciferous isothiocyanates in the diet.

Postharvest treatments such as wounding, ultrasound (US) and the exogenous application of ethylene (ET) and methyl jasmonate (MJ) have been studied as an effective tool to improve the content of secondary metabolites in fresh produce. The present study evaluated the immediate and late response (storage for 72 h at 15 °C) to US treatment (20 min, frequency 24 kHz, amplitude 100 μm) alone and combined with exogenous MJ (250 ppm) and/or ET (1000 ppm) on glucosinolates, isothiocyanates, phenolic compounds and ascorbic acid content in broccoli florets. US treatment increased the extractability of glucosinolates [glucoraphanin (795%), 4-hydroxy glucobrassicin (153%), glucobrassicin (78.6%)] and phenolics [1-sinapoyl-2-feruloylgentiobiose (57.23%)] as compared with the control (CT). The combined application of MJ and US in broccoli florets, induced a synergistic effect on the accumulation of 4-hydroxy glucobrassicin (187.1%), glucoerucin (111.92%), gluconasturtiin (755.9%), neoglucobrassicin (232.8%), 3-O-caffeoylquinic acid (73.4%), 1-sinapoyl-2-ferulolylgentiobiose (56.0%), and 1,2,2-trisinapoylgentiobiose (136.7%) at 72 h of storage. Interestingly, when the three stressors were applied together the synergistic effect of US + MJ observed on the accumulation of glucosinolates and phenolics was repressed. In general, the ascorbic acid content was not affected by US treatment and decreased in most samples during storage. However, when MJ + ET were applied, the content of total ascorbic acid was significantly reduced in CT + MJ + ET and US + MJ + ET samples after 72 h of storage by 53.4% and 86.6%, respectively, as compared with CT 0 h samples. Based on the results herein obtained, the application of US can be an effective tool to enhance the extractability of certain glucocosinolate and phenolic compounds in broccoli. Moreover, due to the synergistic effect observed on the accumulation of bioactive compounds, the combined application of US and MJ could be a practical approach to yield higher levels of glucosinolates and phenolic compounds in broccoli during storage.

Glucoraphanin is one of the best known glucosinolates because of its health benefits. The compound is known to eliminate carcinogens in tissue and hence is frequently studied for its cancer preventative properties. In this work, the total synthesis of α - and β -glucoraphanin epimers was attempted. β -Glu­coraphanin potassium salt was successfully synthesized in high overall yield, whereas the α -epimer was found to be unstable as it decomposed in the final step of the total synthesis. The anti-inflammatory activity of the synthetic glu­co­raphanin was det­ermined by inhibition of the release of tumor necrosis factor alpha (TNF- α ) sec­retion in lipopolysaccharide-stimulated THP-1 cells. It was shown that in the pre­sence of either the synthetic or natural glucoraphanin, TNF- α secretion was sig­nificantly reduced (≈52 % inhibition) at a concen­tration of 15 μM.

Broccoli ( Brassica oleracea var. italica ) is one of exotic vegetables having a high value, highly nutritious, rich source of vitamins, and also it contains the glucoraphanin compound which have anticancerous properties. Integrated nutrient management between organic and inorganic fertilizers is an important demand of the present era to increase broccoli yield and to cultivate a land in such a way that the soil productivity should remain sustainable. This research was conducted to study the effectiveness of organic and inorganic fertilizers on the growth and yield of broccoli. There were eight treatments which consisted of organic and inorganic fertilizers with different applications, single or in combination, and control (without fertilizer). Plant of broccoli as an indicator plant was measured for its height and number of leaves for variables of plant growth parameter, and head weight and head diameter for variables of yield. Application of organic and/or inorganic fertilizers significantly increased the growth of broccoli plant compared with control, but they did not show a significant difference with standard NPK treatment. Combination of organic fertilizer at a rate of 4 tons ha -1 and 50% the standard NPK fertilizer showed the highest diameter and yield of broccoli head. The sole application of organic fertilizer at a rate of 4 tons/ha showed the similargrowth and yield of broccoli with standard NPK treatment.

The temperature effect during autohydrolysis of natural and blanched broccoli was studied. For this purpose, the extraction yield, total phenolic content, antioxidant capacity and glucosinolates type and content of the autohydrolysis liquors have been determined. The development of alginate based hydrogels using the autohydrolysis liquid phase as solvent and the corresponding textural characterisation was also evaluated. In all cases, extraction yields rose with the autohydrolysis temperature up to 150 °C and then decreased. For liquors from natural samples, total phenolic content and antioxidant capacity increased with increasing operation temperature. The opposite trend was identified for blanched samples. Comparable values were obtained at 150 °C for liquors from natural samples and those blanched treated at 100 °C. Glucosinolates content decreased with increasing temperature. Glucoiberin, glucoraphanin, gluconapin, glucobrassicin and gluconasturniin were detected. Hydrogels with suitable textural properties were formulated with the extracted liquors at 100 and 150 °C, independently of the blanching treatment.

Broccoli sprouts are considered functional food as they are naturally enriched in glucoraphanin (GR) that is the biological precursor of the anticancer compound sulforaphane (SFN). Due to its health promoting value, also broccoli sprout juice is becoming very popular. The present study aimed to quantitatively assess the conversion of GR to its hydrolysis products, SFN and SFN-nitrile, during the juice preparation process. We demonstrated that SFN plus SFN-nitrile yield from glucoraphanin is quite low (≈25%) and that some SFN is lost during the juice preparation partially due to the spontaneous conversion to sulforaphane-amine or conjugation to GSH and proteins naturally present in the juice. Our results demonstrate that the detection of the sole SFN free form does not provide reliable information about the real concentration of this functional compound in the juice.

Sulphoraphane originates from glucoraphanin in broccoli and is associated with anti-cancer effects. A preclinical study suggested that daily consumption of broccoli may increase the production of sulphoraphane and sulphoraphane metabolites available for absorption. The objective of this study was to determine whether daily broccoli consumption alters the absorption and metabolism of isothiocyanates derived from broccoli glucosinolates. We conducted a randomised cross-over human study (n18) balanced for BMI and glutathioneS-transferaseμ1 (GSTM1) genotype in which subjects consumed a control diet with no broccoli (NB) for 16 d or the same diet with 200 g of cooked broccoli and 20 g of raw daikon radish daily for 15 d (daily broccoli, DB) and 100 g of broccoli and 10 g of daikon radish on day 16. On day 17, all subjects consumed a meal of 200 g of broccoli and 20 g of daikon radish. Plasma and urine were collected for 24 h and analysed for sulphoraphane and metabolites of sulphoraphane and erucin by triple quadrupole tandem MS. For subjects with BMI >26 kg/m2(median), plasma AUC and urinary excretion rates of total metabolites were higher on the NB diet than on the DB diet, whereas for subjects with BMI 2, plasma AUC and urinary excretion rates were higher on the DB diet than on the NB diet. Daily consumption of broccoli interacted with BMI but notGSTM1genotype to affect plasma concentrations and urinary excretion of glucosinolate-derived compounds believed to confer protection against cancer. This trial was registered as NCT02346812.

The role of epigenetic alterations during cancer has gained increasing attention and has resulted in a paradigm shift in our understanding of mechanisms leading to cancer susceptibility. Sulforaphane (SFN) is a naturally occurring isothiocyanate derived from the precursor glucosinolate, glucoraphanin (GFN), which is found in cruciferous vegetables such as broccoli. Sulforaphane has been shown to suppress tumour growth by several mechanisms including inhibiting histone deacetylases. The objective of this study was to provide a detailed analysis of sulforaphane absorption following a single oral dose of a broccoli sprout supplement in normal dogs. A single dose of broccoli sprout supplement (with active myrosinase) was orally administered to 10 healthy adult dogs. Blood and urine samples were collected prior to dosing, and at various time points post‐dosing. Plasma total SFN metabolite levels peaked at 4 h post‐consumption and were cleared by 24 h post‐consumption. Urinary SFN metabolites peaked at 4 h post‐consumption, and remained detectable at 24 and 48 h post‐supplement consumption. A trend for decrease in histone deacetylase activity was observed at 1 h post‐consumption and a significant decrease was observed at 24 h post‐consumption. The data presented herein indicate that oral SFN is absorbed in dogs, SFN metabolites are detectable in plasma and urine post‐dosing, and SFN and its metabolites have some effect on histone deacetylase activity following a single dose.

Broccoli sprouts are a convenient and rich source of the glucosinolate glucoraphanin, which can generate the chemopreventive agent sulforaphane through the catalytic actions of plant myrosinase or β-thioglucosidases in the gut microflora. Sulforaphane, in turn, is an inducer of cytoprotective enzymes through activation of Nrf2 signaling, and a potent inhibitor of carcinogenesis in multiple murine models. Sulforaphane is also protective in models of diabetes, neurodegenerative disease, and other inflammatory processes, likely reflecting additional actions of Nrf2 and interactions with other signaling pathways. Translating this efficacy into the design and implementation of clinical chemoprevention trials, especially food-based trials, faces numerous challenges including the selection of the source, placebo, and dose as well as standardization of the formulation of the intervention material. Unlike in animals, purified sulforaphane has had very limited use in clinical studies. We have conducted a series of clinical studies and randomized clinical trials to evaluate the effects of composition (glucoraphanin-rich [± myrosinase] vs. sulforaphane-rich or mixture beverages), formulation (beverage vs. tablet) and dose, on the efficacy of these broccoli sprout-based preparations to evaluate safety, pharmacokinetics, pharmacodynamic action, and clinical benefit. While the challenges for the evaluation of broccoli sprouts in clinical trials are themselves formidable, further hurdles must be overcome to bring this science to public health action.

Glucoraphanin is a major secondary metabolite found in Brassicaceae vegetables, especially broccoli, and its degradation product sulforaphane plays an essential role in anticancer. The fine mapping of sulforaphane metabolism quantitative trait loci (QTLs) in broccoli florets is necessary for future marker-assisted selection strategies. In this study, we utilized a doubled haploid population consisting of 176 lines derived from two inbred lines (86,101 and 90,196) with significant differences in sulforaphane content, coupled with extensive genotypic and phenotypic data from two independent environments. A linkage map consisting of 438 simple sequence repeats markers was constructed, covering a length of 1168.26 cM. A total of 18 QTLs for sulforaphane metabolism in broccoli florets were detected, 10 were detected in 2017, and the other 8 were detected in 2018. The LOD values of all QTLs ranged from 3.06 to 14.47, explaining 1.74–7.03% of the biochemical variation between two years. Finally, 6 QTLs (qSF-C3-1, qSF-C3-2, qSF-C3-3, qSF-C3-5, qSF-C3-6 and qSF-C7) were stably detected in more than one environment, each accounting for 4.54–7.03% of the phenotypic variation explained (PVE) and a total of 30.88–34.86% of PVE. Our study provides new insights into sulforaphane metabolism in broccoli florets and marker-assisted selection breeding in Brassica oleracea crops.

COVID-19 caused by a positive-sense single stranded RNA virus named as severe acute respiratory syndrome-Coronavirus-2 (SARS-CoV-2) triggered the global pandemic. This virus has infected about 10.37 Crores and taken lives of 2.24 Crores people of 213 countries to date. To cope-up this emergency clinical trials are undergoing with some existing drugs like remdesivir, flavipiravir, lopinavir-ritonavir, nafamostat, doxycycline, hydroxy-chloroquine, dexamethasone, etc., despite their severe toxicity and health hazards among diabetics, hypertensive, cardiac patients or normal individuals. The lack of safe and approved treatment for COVID-19 has forced the scientific community to find novel and safe compounds with potential efficacy. This study evaluates a few selective herbal compounds like glucoraphanin, vitexin, niazinin, etc., as a potential inhibitor of the spike protein and 3-chymotrypsin-like protease (3CLpro) or main protease (Mpro) of SARS-COV-2 through in-silico virtual studies such as molecular docking, target analysis, toxicity prediction and ADME prediction and supported by a Molecular-Dynamic simulation. Selective phytocompounds were docked successfully in the binding site of spike glycoprotein and 3CLpro (Mpro) of SARS-CoV-2. In-silico approaches also predict this molecule to have good solubility, pharmacodynamic property and target accuracy through MD simulation and ADME studies. These hit molecules niazinin, vitexin, glucoraphanin also obey Lipinski's rule along with their stable binding towards target protein of the virus, which makes them suitable for further biochemical and cell-based assays followed by clinical investigations to highlight their potential use in COVID-19 treatment.Communicated by Ramaswamy H. Sarma.

A large proportion of broccoli biomass is lost during primary production, distribution, processing, and consumption. This biomass is rich in polyphenols and glucosinolates and can be used for the production of bioactive rich ingredients for food and nutraceutical applications. This study evaluated thermosonication (TS) (18 kHz, 0.6 W/g, 40–60 °C, 3–7 min) for the pre-treatment of broccoli florets to enhance enzymatic conversion of glucoraphanin into the bioactive sulforaphane. TS significantly increased sulforaphane yield, despite a decrease in myrosinase activity with increasing treatment intensity. The highest sulforaphane yield of ~2.9 times that of untreated broccoli was observed for broccoli thermosonicated for 7 min at 60 °C, which was 15.8% higher than the corresponding yield for thermal processing without sonication (TP) at the same condition. This was accompanied by increase in the residual level of glucoraphanin (~1.8 and 2.3 time respectively after TP and TS at 60 °C for 7 min compared to control samples) indicating that treatment-induced release of bound glucoraphanin from the cell wall matrix and improved accessibility could be at least partially responsible for the enhanced sulforaphane yield. The result indicates the potential of TS for the conversion of broccoli biomass into high sulforaphane broccoli-based ingredients.

Glucosinolates are biologically active secondary metabolites in Brassicaceae plants that play a critical role in positive and negative interactions between plants and root-associated microbial communities. The aim of this study was to develop a reversed-phase liquid chromatography method to quantify and identify intact glucosinolates in the root of Arabidopsis thaliana (Arabidopsis) grown in non-sterile natural soil by using liquid chromatography-hybrid triple quadruple-linear ion trap (LC-QqQ(LIT)) mass spectrometry. The Synergi Fusion C18-based column was found to be effective for sufficient retention and separation of nine intact glucosinolates without the need for time-consuming desulfation or ion-pairing steps. Method validation results showed satisfactory inter-day and intra-day precision for all glucosinolates except for 4-hydroxyglucobrassicin. Good inter-day and intra-day accuracy and recovery results were observed for glucoiberin, gluconapin, glucobrassicin, 4-methoxyglucobrassicin and neoglucobrassicin. However, for 4-hydroxyglucobrassicin, glucoraphanin and glucoerucin corrections to quantification results might be necessary since the recovery and accuracy results were not optimal. Matrix effects were shown to have a negligible effect on the ionization of all target analytes. The established liquid chromatography&ndash, tandem mass spectrometry (LC-MS/MS) method was applied to quantify target intact glucosinolates in Arabidopsis root crude extract of four different wild-type accessions where differences in terms of concentration and composition of intact glucosinolates were observed. Employment of sensitive and selective precursor ion survey scan of m/z 97 in combination with the information-dependent acquisition (IDA) of the enhanced product ion (EPI) dependent scan (Prec97-IDA-EPI) using LC-QqQ(LIT) provided high confidence in structural characterization of diverse intact glucosinolate profiles in complex Arabidopsis root crude extract.

International audience; COVID-19 is described in a clinical case involving a patient who proposed the hypothesis that Nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-interacting nutrients may help to prevent severe COVID-19 symptoms. Capsules of broccoli seeds containing glucoraphanin were being taken before the onset of SARS-CoV-2 infection and were continued daily for over a month after the first COVID-19 symptoms. They were found to reduce many of the symptoms rapidly and for a duration of 6-12 h by repeated dosing. When the patient was stable but still suffering from cough and nasal obstruction when not taking the broccoli capsules, a double-blind induced cough challenge confirmed the speed of onset of the capsules (less than 10 min). A second clinical case with lower broccoli doses carried out during the cytokine storm confirmed the clinical benefits already observed. A third clinical case showed similar effects at the onset of symptoms. In the first clinical trial, we used a dose of under 600 μmol per day of glucoraphanin. However, such a high dose may induce pharmacologic effects that require careful examination before the performance of any study. It is likely that the fast onset of action is mediated through the TRPA1 channel. These experimental clinical cases represent a proof-of-concept confirming the hypothesis that Nrf2-interacting nutrients are effective in COVID-19. However, this cannot be used in practice before the availability of further safety data, and confirmation is necessary through proper trials on efficacy and safety.