Method validation, residue behaviour and dietary risk assessment of insecticides (cyantraniliprole, acetamiprid, flubendiamide and its metabolite, des-iodo flubendiamide) in or on broccoli using LC-MS/MS.

Residue behaviour and dietary risk assessment of cyantraniliprole, flubendiamide and acetamiprid in broccoli were carried out using the QuEChERS (quick, easy, cheap, effective, rugged and safe) technique coupled with LC-MS/MS. The QuEChERS technique was validated on parameters such as linearity, accuracy, precision, robustness, matrix effects, limit of quantification (LOQ), specificity, retention time and ion ratio as per SANTE (Directorate General for Health and Food Safety) guidelines to attest to the specificity, accuracy and precision of the analytical method in estimating insecticide residues in and on broccoli heads and cropped soil. The LOQ of the method for all three insecticides was 0.01 mg/kg. The initial deposits of cyantraniliprole, flubendiamide and acetamiprid reduced to half of its concentration in 1.873-2.354, 1.975-2.484 and 1.371-1.620 days, respectively. No residues were detected in broccoli-cropped soil at harvest time (30 days after last spray). The proposed maximum residue limits (MRLs) of 1.5, 0.5-0.9 and 2.0-3 mg/kg for cyantraniliprole, flubendiamide and acetamiprid were calculated using the Organisation for Economic Co-operation and Development MRL calculator. The acute and chronic dietary risk assessment of the tested insecticides identified no appreciable dietary risk to the Indian population from the consumption of broccoli heads. The findings of no dietary risk highlight the importance of informed pesticide usage in broccoli and the proposed MRL derived from this study offers crucial guidelines for the regulatory authorities, ensuring the safety of broccoli consumption.

Potential therapeutic properties of broccoli extract and soy isoflavones on improvement endometriosis and involved oxidative parameters.

OBJECTIVES: In Endometriosis is a gynecological disorder characterized by the growth of endometrial tissue outside the uterine cavity that is associated with chronic pelvic pain and subfertility. The purpose of the study was to investigate the effect of broccoli extract (BE) alone and in combination with soy isoflavones (SI) on endometrial implants in female rat. METHODS: In this study, endometriosis was induced surgically in 40 mature female rats. The rats were divided into 5 groups that were treated by oral gavage for 6 weeks with 0.5 mL of saline 0.9 %/day (control group), BE (3,000 mg/kg/day), SI (50 mg/kg/day), BE/soy isoflavones (BE 3000 mg/kg/day + soy isoflavones 50 mg/kg/day) and diphereline as a standard medication (3 mg/kg) intramuscularly. At the end of treatments, the volume and histopathology of the endometrial implants were compared among the 5 groups. The serum levels of oxidative parameters including superoxide dismutase (SOD), malondialdehyde (MDA) and tumor necrosis factor alpha (TNF-α) were also compared between the groups. The volume of the implants significantly decreased in diphereline group (p=0.002). RESULTS: The histopathological grade of endometrial implants in BE/SI and diphereline group were significantly decreased compared to the control group (p=0.001). The serum levels of SOD in BE group were enhanced significantly in comparison to the control group (p=0.034). CONCLUSIONS: BE in combination with SI decreased the growth and histopathologic grades of transplanted endometrial implants. These herbal compounds may have the potential therapeutic effect to be used as an alternative medication for the treatment of endometriosis.

Endometriosis happens when tissue similar to the uterus tissue grows outside of the uterus. It can be associated with severe pelvic pain and make several difficulties to get pregnant. Since the use of hormonal drugs is prohibited when planning to get pregnant and due to the common recurrence of lesions after surgery, scientists have been encouraged to study regarding the efficacies of natural products on improvement of endometriosis. In this study, it is represented that the dietary consumption of broccoli extract combined with soy isoflavones may have potential beneficial properties on clinical symptoms of endometriosis.

The Effect of Broccoli Glucosinolates Hydrolysis Products on Botrytis cinerea: A Potential New Antifungal Agent.

The present study investigates the interactions between eight glucosinolate hydrolysis products (GHPs) sourced from broccoli by-products and the detoxifying enzymes of Botrytis cinerea, namely eburicol 14-alpha-demethylase (CYP51) and glutathione-S-transferase (GST), through in silico analysis. Additionally, in vitro assays were conducted to explore the impact of these compounds on fungal growth. Our findings reveal that GHPs exhibit greater efficacy in inhibiting conidia germination compared to mycelium growth. Furthermore, the results demonstrate the antifungal activity of glucosinolate hydrolysis products derived from various parts of the broccoli plant, including inflorescences, leaves, and stems, against B. cinerea. Importantly, the results suggest that these hydrolysis products interact with the detoxifying enzymes of the fungus, potentially contributing to their antifungal properties. Extracts rich in GHPs, particularly iberin and indole-GHPs, derived from broccoli by-products emerge as promising candidates for biofungicidal applications, offering a sustainable and novel approach to plant protection by harnessing bioactive compounds from agricultural residues.

Involvement of glucosinolates and phenolics in the promotion of broccoli seedling growth through the modulation of primary and secondary metabolism.

Secondary metabolites play an essential role in plant defense. However, the role of glucosinolates and phenols in brassica crop yield in the context of environmentally friendly agricultural practices has not been established. Our study investigated the effects of a Brassica extract, rich in these metabolites, on the physiology and metabolism of broccoli (Brassica oleracea L. var. italica) seedlings and the subsequent development of the plants in adult stages. The results showed an increase in growth in the extract-treated seedlings, which was associated with an alteration of primary and secondary metabolism. In particular, there was an increase in the levels of amino acids, phenolic compounds and hormones, while the levels of glucosinolates decreased. Lipid peroxidation diminished in treated plants, indicating improved membrane integrity. Treated plants subsequently grown in hydroponically showed increased water use efficiency, transpiration, and internal carbon, which contributed to the improved growth of these plants. Overall, our findings underscore the potential of the glucosinolates and phenols ratio as essential to improve crop growth and stress tolerance, as well as revealed the interest of studying the mechanisms involved in the possible uptake and integration of GSLs by broccoli seedlings after external application.

One-Step Electrochemically Prepared Bionic Hierarchical Nickel Black@Graphene Composite Membrane for Desalination by Solar-Thermal Energy Conversion.

Ingenious microstructure construction and appropriate composition selection are effective strategies for achieving enhanced performance of photothermal materials. Herein, a broccoli-like hierarchical nickel black@graphene (Ni@Gr) membrane for solar-driven desalination was prepared by a one-step electrochemical method, which was carried out simultaneously with the electrochemical exfoliation of graphene and the co-deposition of Ni@Gr material. The bionic hierarchical structure and the chemical composition of the Ni@Gr membrane increased the sunlight absorption (90.36%) by the light-trapping effect and the introduction of graphene. The Ni@Gr membrane achieved high evaporation rates of 2.05 and 1.16 kg m-2 h-1 under simulated (1 sun) and outdoor sunlight conditions, respectively. The superhydrophilicity and the hierarchical structure of the Ni@Gr membrane jointly reduced the evaporation enthalpy (1343.6 kJ/kg), which was beneficial to break the theoretical limit of the evaporation rate (1.47 kg m-2 h-1). This work encourages the application of bionic metal-carbon composite photothermal materials in solar water evaporation.

Transcriptome profiling reveals key regulatory factors and metabolic pathways associated with curd formation and development in broccoli.

Broccoli, a cruciferous vegetable, has a unique indeterminate inflorescence structure known as curds. It is the main edible organ of broccoli and has a rich nutritional value and health benefits. However, the formation and development mechanism of the curd is still not well understood. In the present study, the shoot apical meristem (SAM) stage and three different development stages of curd (formation stage (FS), expansion stage (ES), and maturation stage (MS)) were identified and subjected to transcriptome sequencing to uncover the potential genes and regulatory networks involved in curd formation and development. The results indicated that the genes associated with the development of SAM such as BolAP1A, BolAP1C, BolCAL, and BolAGL6 play an important role in the abnormal differentiation of the curd apical buds. The genes, BolFRI, BolbHLH89, BolKAN4, BolAGL12, and BolAGL24, displayed significantly differential expression patterns in curd development may function in the regulation of the transition from inflorescence meristem (IM) to floral meristem (FM). Moreover, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the differentially expressed genes (DEGs) indicate that phytohormones, such as auxin (AUX), gibberellins (GA), and abscisic acid (ABA) also play an important role in SAM proliferation and the transition from SAM to IM. In addition, the genes regulating photosynthetic reaction (BolLHCA1, BolLHCB1, BolPsbO, etc.) have a key involvement in the differentiation of secondary IMs during curd expansion. The genes associated with the metabolism of starch and sucrose (e.g., BolSPS4, BolBAM4) were significantly upregulated at the MS should contribute to the maturation of the curd. These findings provide new insights into the potential key regulatory factors and metabolic pathways involved in the formation and development of broccoli curds.

Advances in and Perspectives on Transgenic Technology and CRISPR-Cas9 Gene Editing in Broccoli.

Broccoli, a popular international Brassica oleracea crop, is an important export vegetable in China. Broccoli is not only rich in protein, vitamins, and minerals but also has anticancer and antiviral activities. Recently, an Agrobacterium-mediated transformation system has been established and optimized in broccoli, and transgenic transformation and CRISPR-Cas9 gene editing techniques have been applied to improve broccoli quality, postharvest shelf life, glucoraphanin accumulation, and disease and stress resistance, among other factors. The construction and application of genetic transformation technology systems have led to rapid development in broccoli worldwide, which is also good for functional gene identification of some potential traits in broccoli. This review comprehensively summarizes the progress in transgenic technology and CRISPR-Cas9 gene editing for broccoli over the past four decades. Moreover, it explores the potential for future integration of digital and smart technologies into genetic transformation processes, thus demonstrating the promise of even more sophisticated and targeted crop improvements. As the field continues to evolve, these innovations are expected to play a pivotal role in the sustainable production of broccoli and the enhancement of its nutritional and health benefits.

Characterization of selenium-containing broccoli (Brassica oleracea L. var. italica planch) proteins and evaluation of antioxidant activity by electron spin resonance.

Selenoproteins found in selenium (Se)-enriched vegetables play a vital role in maintaining human health. In this study, four Se-containing broccoli proteins (Se-BP: albumin, globulin, prolamin, and glutelin) were continuous extracted by Osborne method. Three ultrafiltered fractions were subsequently obtained from the glutelin hydrolysate, composed of Se-contained broccoli peptides (Se-Bp) with different molecular weights (MW), namely, < 1 kDa, 1-3 kDa, and 3-10 kDa. Glutelin exhibited the highest protein yield (65.60 ± 1.07%), purity (78.39 ± 0.95%), nutritional value, organic Se content (88.05 ± 0.32% of total Se content), and Se speciation distribution (selenocystine, selenomethionine, methylselenocysteine, and selenoethionine). Additionally, the antioxidant activity of different MW of Se-Bp was assessed using electron spin resonance spectroscopy. The results revealed that antioxidant activity of the candidate peptide is dependent upon its Se content, amino acid composition, and MW, especially Se-Bp with MW of 1-3 kDa displayed the strongest free radical scavenging ability.

Exploring fermentation with lactic acid bacteria as a pretreatment for enhancing antioxidant potential in broccoli stem powders.

Fruit and vegetable industries face a major environmental challenge with food loss and waste. Broccoli stems, comprising 38% of the plant's total weight, are usually discarded by the industry producing fourth-range and ready-to-use products, despite being rich in antioxidants, vitamins, fiber, carotenoids, phenolic compounds, and glucosinolates. Addressing the challenge of reducing waste in this sector includes the production of stable and nutrient-concentrated powders, which can be consumed directly or used as ingredients in functional food formulation. This study investigated fermentation with lactic acid bacteria (Limosilactobacillus reuteri, Lactiplantibacillus plantarum, and Lactobacillus salivarius) as a pretreatment for enhancing antioxidant and probiotic potential in broccoli stem powders. Results showed maximum counts 24 h after inoculation, and no effect of the previous disruption intensity on microbial growth was observed. Fermenting broccoli stems for 24 h with the three microbial strains led to a significant increase in total phenols and flavonoids but to a general reduction in the samples' capacity to scavenge DPPH and ABTS free radicals. Overall, ground broccoli stems exhibited the most favorable antioxidant properties following the 24 h fermentation step. The subsequent freeze-drying and final grinding had minimal impact on the microbial population but significantly enhanced the extractability of the antioxidant compounds. This study offers a valuable reference for researchers and stakeholders exploring the development of new products and innovations from vegetable waste.

Quantitative Determination of Biogenic Element Contents and Phytochemicals of Broccoli (Brassica oleracea var. italica) Cooked Using Different Techniques.

In this study, the effect of different cooking techniques on broccoli moisture, total phenolic, total flavonoid, and radical scavenging capacity results, polyphenol contents, and their quantitative values was investigated. The total phenolic quantities of fresh and cooked broccoli samples were assessed to be between 36.32 (conventional boiling) and 423.39 mg GAE/100 g (microwave heating). The radical scavenging activities of the broccoli samples were reported between 2.55 (conventional boiling) and 4.99 mmol/kg (microwave heating). In addition, catechin and rutin quantities of the fresh and cooked broccoli samples were measured to be between 2.24 (conventional boiling) and 54.48 mg/100 g (microwave heating), and between 0.55 (conventional boiling) and 16.33 mg/100 g (microwave heating), respectively. The most abundant elements in fresh and cooked broccoli samples were K, Ca, P, S, and Mg. The results showed some changes depending on cooking techniques compared to the control. The bioactive properties of broccoli samples cooked by means of conventional boiling, boiling in vacuum bag, and high-pressure boiling were established to be lower compared to the fresh sample. Catechin, 3,4-dihydroxybenzoic acid, rutin, and gallic acid were the key phenolic compounds of fresh and cooked broccoli samples. The phenolic components of broccoli were significantly affected by the applied cooking techniques. The highest protein in broccoli samples was determined in the broccoli sample cooked by boiling in a vacuum bag. There were statistically significant changes among the mineral results of broccoli cooked with different cooking methods.

Development and Application of a Selective Analytical Method for Indole Metabolites in Urine: Dietary Exposure Biomarkers for Broccoli Consumption.

The identification of dietary exposure biomarkers is crucial for advancing our understanding of the health benefits of specific foods. Broccoli, a vegetable with well-known anticancer properties, contains active ingredients, such as isothiocyanates with indole side chains. Hence, indole metabolites related to broccoli consumption have the potential to serve as biomarkers of dietary exposure. In this work, we developed a new analytical method for indole metabolites in urine using a poly(deep eutectic solvents)-molecularly imprinted polymer/vinyl-functionalized graphene oxide (PDESs-MIP/VGO) in miniaturized centrifugal pipet-tip solid-phase extraction (CPT-SPE) coupled with liquid chromatography. This method integrates the strengths of PDESs-MIP/VGO, including rich adsorption interactions, high adsorption capacity, and excellent selectivity, with the simplicity and cost-effectiveness of CPT-SPE. The proposed method demonstrated low limits of quantification (1.2-2.5 ng mL-1), high accuracy (91.7-104.8%), and good precision (relative standard deviation ≤4.4%). By applying this method to analyze indole metabolites in urine, our results suggested that indole-3-carbinol and indole-3-acetonitrile have the potential to emerge as reliable dietary exposure biomarkers for broccoli intake. Furthermore, highly selective analytical methods based on molecular imprinting technology are advantageous for precise screening and analysis of dietary exposure biomarkers associated with food consumption.

Evaluation of the influence of N-acetylcysteine and broccoli extract on systemic paraquat poisoning: Implications for biochemical, physiological, and histopathological parameters in rats.

Objectives: Paraquat (PQ), a potent environmental herbicide, is recognized for inducing irreparable toxic damage to biological systems. This study aimed to evaluate the effectiveness of N-acetylcysteine (NAC) and broccoli extract, individually and in combination, in alleviating PQ poisoning in rats, leveraging the exceptional anti-oxidant, anti-inflammatory, and anti-apoptotic properties of broccoli. Materials and Methods: Seventy Wistar rats were categorized into seven groups: C (control, vehicle), PQ (paraquat at 40 mg/kg), BC (broccoli extract at 300 mg/kg), NC (N-acetylcysteine at the same dose of 300 mg/kg), and combined groups PQ+BC, PQ+NC, and NC+PQ+BC, all administered equivalent doses. After 42 days, blood samples were collected to evaluate liver and kidney parameters, proinflammatory biomarkers, caspase-3, and caspase-9. Lung tissues were excised, with one part preserved for hydroxyproline and oxidative stress parameter measurement and another sectioned and stained for histopathological analysis. Results: The PQ group exhibited the highest lung-to-body weight (LW/BW) ratio, while the PQ+BC+NC group demonstrated the lowest ratio. Results indicated an elevated lung hydroxyproline concentration and a significant reduction in anti-oxidant enzymes (catalase, glutathione peroxidase, superoxide dismutase, and total anti-oxidant capacity) (P<0.001). The PQ+BC group showed modified malondialdehyde levels, reaching a peak in the PQ group. Additionally, a significant decrease in tumor necrosis factor, interleukin-1, caspase-3, and caspase-9 was observed in the PQ+BC+NC group (P<0.01). Pulmonary edema, hyperemia, and severe hemorrhage observed in the PQ group were notably reduced in the PQ+BC+NC group. Conclusion: The combination of active compounds from broccoli and NAC demonstrated significant systemic and pulmonary effects in mitigating PQ-induced toxicity.

How long can you store vitamins? Stability of tocopherols and tocotrienol during different storage conditions in broccoli and blueberries.

Differences between the stability of α-, ß-, γ-, and δ-tocopherol as well α-tocotrienol stored at -20 °C and -80 °C were studied in broccoli and blueberry samples. Before storage up to 28 days, they underwent different initializing processes such as freezing quickly with liquid nitrogen and freeze-drying, followed by homogenization. While α-tocopherol levels in blueberries did not significantly differ, levels in broccoli were substantially higher after homogenization of freeze-dried samples compared to fresh broccoli samples. This might be caused by higher extractability of α-tocopherol from the changed cell structure. Storage of fresh broccoli samples at -20 °C led to decreasing α-tocopherol levels. Nevertheless, the deviation between freeze-dried samples to the initial fresh samples and fresh samples frozen with liquid nitrogen stored at -20 °C for 7 days were in the same order of magnitude. In conclusion, storage up to 7 days for vitamin relevant samples before analysis seemed to be justifiable.

The Effect of Broccoli Glucoraphanin Supplementation on Ameliorating High-Fat-Diet-Induced Obesity through the Gut Microbiome and Metabolome Interface.

SCOPE: Obesity and its metabolic comorbidities pose a major global challenge for public health. Glucoraphanin (GRN) is a natural bioactive compound enriched in broccoli that is known to have potential health benefits against various human chronic diseases. METHODS AND RESULTS: This study investigats the effects of broccoli GRN supplementation on body weight, metabolic parameters, gut microbiome and metabolome associated with obesity. The study is conducted on an obese-related C57BL/6J mouse model through the treatment of normal control diet, high-fat diet (HFD)and GRN-supplemented HFD (HFD-GRN) to determine the metabolic protection of GRN. The results shows that GRN treatment alleviates obesity-related traits leading to improved glucose metabolism in HFD-fed animals. Mechanically, the study noticed that GRN significantly shifts the gut microbial diversity and composition to an eubiosis status. GRN supplement also significantly alters plasma metabolite profiles. Further integrated analysis reveal a complex interaction between the gut microbes and host metabolism that may contribute to GRN-induced beneficial effects against HFD. CONCLUSION: These results indicate that beneficial effects of broccoli GRN on reversing HFD-induced adverse metabolic parameters may be attributed to its impacts on reprogramming microbial community and metabolites. Identification of the mechanistic functions of GRN further warrants it as a dietary candidate for obesity prevention.

Formation of DNA Adducts by 1-Methoxy-3-indolylmethylalcohol, a Breakdown Product of a Glucosinolate, in the Mouse: Impact of the SULT1A1 Status-Wild-Type, Knockout or Humanised.

We previously found that feeding rats with broccoli or cauliflower leads to the formation of characteristic DNA adducts in the liver, intestine and various other tissues. We identified the critical substances in the plants as 1-methoxy-3-indolylmethyl (1-MIM) glucosinolate and its degradation product 1-MIM-OH. DNA adduct formation and the mutagenicity of 1-MIM-OH in cell models were drastically enhanced when human sulfotransferase (SULT) 1A1 was expressed. The aim of this study was to clarify the role of SULT1A1 in DNA adduct formation by 1-MIM-OH in mouse tissues in vivo. Furthermore, we compared the endogenous mouse Sult1a1 and transgenic human SULT1A1 in the activation of 1-MIM-OH using genetically modified mouse strains. We orally treated male wild-type (wt) and Sult1a1-knockout (ko) mice, as well as corresponding lines carrying the human SULT1A1-SULT1A2 gene cluster (tg and ko-tg), with 1-MIM-OH. N2-(1-MIM)-dG and N6-(1-MIM)-dA adducts in DNA were analysed using isotope-dilution UPLC-MS/MS. In the liver, caecum and colon adducts were abundant in mice expressing mouse and/or human SULT1A1, but were drastically reduced in ko mice (1.2-10.6% of wt). In the kidney and small intestine, adduct levels were high in mice carrying human SULT1A1-SULT1A2 genes, but low in wt and ko mice (1.8-6.3% of tg-ko). In bone marrow, adduct levels were very low, independently of the SULT1A1 status. In the stomach, they were high in all four lines. Thus, adduct formation was primarily controlled by SULT1A1 in five out of seven tissues studied, with a strong impact of differences in the tissue distribution of mouse and human SULT1A1. The behaviour of 1-MIM-OH in these models (levels and tissue distribution of DNA adducts; impact of SULTs) was similar to that of methyleugenol, classified as "probably carcinogenic to humans". Thus, there is a need to test 1-MIM-OH for carcinogenicity in animal models and to study its adduct formation in humans consuming brassicaceous foodstuff.

Dietary plant microRNAs as potential regulators of cellular cholesterol efflux.

AIM: Epidemiological evidence suggests adherence to vegetable-rich diets is associated to atheroprotective effects and bioactive components are most likely to play a relevant role. The notion of inter-kingdom regulation has opened a new research paradigm and perhaps microRNAs (miRNAs) from edible vegetables could influence consumer gene expression and lead to biological effects. We aimed to investigate the potential impact of broccoli-derived miRNAs on cellular cholesterol efflux in vitro. METHODS: Four miRNAs (miR159a, miR159b, miR166a and miR403) from Brassica oleracea var. italica (broccoli), a widely consumed cruciferous vegetable, were selected for further investigation, based on their high abundancy in this vegetable and their presence in other plants. Selected miRNAs were synthesized with a 3'-terminal 2'-O-methylation and their cellular toxicity, in vitro gastrointestinal resistance and cellular uptake were evaluated. Potential target genes within the mammalian transcriptome were assessed in silico following pathway analysis. In vitro cholesterol efflux was assessed in human THP-1-derived macrophages. RESULTS: miRNAs survival to in vitro GI digestion was around 1%, although some variation was seen between the four candidates. Cellular uptake by mammalian cells was confirmed, and an increase in cholesterol efflux was observed. Pathway analysis suggested these miRNAs are involved in biological processes related to phosphorylation, phosphatidylinositol and Wnt signaling, and to the insulin/IGF pathway. CONCLUSIONS: Health-promoting properties attributed to cruciferous vegetables, might be mediated (at least in part) through miRNA-related mechanisms.

Unravelling Glucoraphanin and Glucoerucin Metabolism across Broccoli Sprout Development: Insights from Metabolite and Transcriptome Analysis.

Variations in the concentration of glucoraphanin (GRA) and glucoerucin (GER), as well as the corresponding breakdown products, isothiocyanates (ITCs) and nitriles, were investigated during the growth of broccoli sprouts. The concentrations of GRA and GER decreased sharply from 33.66 µmol/g to 11.48 µmol/g and 12.98 µmol/g to 8.23 µmol/g, respectively, after seed germination. From the third to the seventh day, both GRA and GER were maintained as relatively stable. The highest concentrations of sulforaphane (17.16 µmol/g) and erucin (12.26 µmol/g) were observed on the first day. Hereafter, the concentrations of nitrile hydrolyzed from GRA or GER were higher than those of the corresponding ITCs. Moreover, the ratio of sulforaphane to sulforaphane nitrile decreased from 1.35 to 0.164 from 1 d to 5 d, with a similar trend exhibited for erucin/erucin nitrile after 2 d. RNA-seq analysis showed that BolMYB28 and BolCYP83A1, involved in aliphatic glucosinolate (GSL) biosynthesis, remained largely unexpressed until the third day. In contrast, the genes operating within the GSL-myrosinase hydrolysis pathway were highly expressed right from the beginning, with their expression levels increasing significantly after the third day. Additionally, we identified two BolESPs and six BolNSPs that might play important roles in promoting the production of nitriles during the development of broccoli sprouts.

Benign and malignant prolapsed uterine tumors : 4 case reports of an extremely rare entities.

Prolapsed uterine tumors within the cervix or vagina, are attached to the uterine cavity by a soft tissue stalk. Malignant tumors and leiomyoma are the first diagnostic considerations for a prolapsed uterine mass with a visible stalk at MRI. This article describes 4 cases of patients who presented with large, necrotic prolapsed uterine tumors that were surgically confirmed and were diagnosed prospectively on the basis of MRI findings. Imaging, particularly MRI, plays a crucial role in the management of patients with prolapsed pedunculated uterine tumors, especially for pre-operative localization and surgical treatment. The 4 patients underwent a total hysterectomy with a complete resection of the mass. The histopathological report confirmed in the first case the diagnosis of a uterine leiomyoma with aseptic necrobiosis, in the second and third cases a sarcoma, and in the fourth case a serous adenocarcinoma.

Aggressive Alternaria brassicicola with Reduced Fungicide Sensitivity Can Be Associated with Naturally Infested Broccoli Seeds.

Alternaria brassicicola is a part of the Alternaria complex that causes leaf blight and head rot (ABHR) in brassica crops. Infested broccoli seeds can play an important role in introducing A. brassicicola in transplant houses and production fields. However, characterization of natural seed infestation and seed-to-seedling transmission of A. brassicicola in broccoli is yet to be demonstrated. In this research, we characterized Alternaria spp. isolates from commercial broccoli seedlots for their species identity, pathogenicity, and aggressiveness on broccoli and their sensitivity to a quinone-outside inhibitor (QoI) fungicide (azoxystrobin). Two hundred commercial seedlots from two broccoli cultivars, Cultivar 1 (EC; n = 100 seedlots) and Cultivar 2 (ED; n = 100 seedlots) were, evaluated for the presence of A. brassicicola under in vitro conditions using a seedling grow-out assay. Alternaria spp. was detected in 31 and 28% of the commercial seedlots of Cultivar 1 and Cultivar 2, respectively. The seed-to-seedling transmission (%) varied considerably within each positive-infested seedlot, which ranged from 1.3 to 17.3%. Subsequent molecular identification of single-spore cultures (n = 138) was made by sequencing four housekeeping genes: actin, the major allergen (Alta1), plasma membrane ATPase, and glyceraldehyde-3-phosphate dehydrogenase (GPD), and the sequences were concatenated and compared for the phylogenetic distance with diverse Alternaria species. Ninety-six percent (n = 133) of the isolates formed a cluster with a known A. brassicicola based on a multigene phylogeny, which were later confirmed as A. brassicicola using a species-specific PCR assay. One hundred percent of the A. brassicicola seed isolates (n = 133) were either highly or moderately aggressive on broccoli (cultivar Emerald Crown) based on a detached leaf assay. Sensitivity of representative A. brassicicola isolates (n = 58) to azoxystrobin was evaluated using a spore germination assay, and the EC50 values (effective fungicide concentration [ppm] at which germination of conidia of isolates were reduced by 50% compared to control) for each isolate was determined. A. brassicicola isolates from naturally infested commercial broccoli seeds were sensitive to azoxystrobin with considerably low EC50 values in the range of <0.0001 to 0.33 ppm; however, there were a few isolates (14%) that showed 100-fold reduced sensitivity from the most sensitive isolate (EC50 = 0.0001 ppm). Our results confirm that commercial broccoli seedlots can be naturally contaminated with pathogenic and aggressive A. brassicicola. We also provide evidence for the potential presence of A. brassicicola isolates with reduced azoxystrobin-sensitivity in naturally infested commercial broccoli seedlots, which has never been reported before. Together, these findings may have implications in considerations for seed-health testing, seed treatments, and greenhouse scouting to limit introduction of infested seedlots in commercial broccoli fields.

The synthesis of broccoli sprout extract-loaded silk fibroin nanoparticles as efficient drug delivery vehicles: development and characterization.

Targeted drug delivery of biological molecules using the development of biocompatible, non-toxic and biodegradable nanocarriers can be a promising method for cancer therapy. In this study, silk fibroin protein nanoparticles (SFPNPs) were synthesized as a targeted delivery system for sulforaphane-rich broccoli sprout extract (BSE). The BSE-loaded SFPNPs were conjugated with polyethylene glycol and folic acid, and then their physicochemical properties were characterized via UV-Vis, XRD, FTIR, DLS, FE-SEM and EDX analyses. In vitro, the release profile, antioxidant and anticancer activities of NPs were also studied. The FE-SEM and DLS analyses indicated stable NPs with an average size of 88.5 nm and high zeta potential (-32 mV). The sulforaphane release profile from NPs was pH-dependent, with the maximum release value (70%) observed in simulated intestinal fluid (pH = 7.4). Encapsulation of BSE also decreased the release rate of sulforaphane from the capsules compared to free BSE. In vitro cytotoxicity of BSE and NPs on breast cancer cell lines (MCF-7) was concentration-dependent, and the IC50 for BSE and NPs were 54 and 210 µg ml-1, respectively. Moreover, the NPs demonstrated no appreciable cytotoxicity in normal mouse fibroblast (L929) cell lines. These results indicated that biocompatible NPs synthesized as controlled and long-term targeted drug delivery systems can be a potential candidate for breast cancer therapy.