Characterization, biological activity, and anticancer effect of green-synthesized gold nanoparticles using Nasturtium officinale L.

BACKGROUND: Nanostructured materials used have unique properties and many uses in nanotechnology. The most striking of these is using herbal compounds for the green synthesis of nanoparticles. Among the nanoparticle types used for green synthesis, gold nanoparticles (AuNPs) are used for cancer therapy due to their stable structure and non-cytotoxic. Lung cancer is the most common and most dangerous cancer worldwide in terms of survival and prognosis. In this study, Nasturtium officinale (L.) extract (NO), which contains biomolecules with antioxidant and anticancer effects, was used to biosynthesize AuNPs, and after their characterization, the effect of the green-synthesized AuNPs against lung cancer was evaluated in vitro. METHODS: Ultraviolet‒visible (UV‒Vis) spectrophotometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDS), multiple analysis platform (MAP), and Fourier transform infrared (FT-IR) spectroscopy analyses were performed to characterize the AuNPs prepared from the N. officinale plant extract. Moreover, the antioxidant activity, total phenolic and flavonoid contents and DNA interactions were examined. Additionally, A549 lung cancer cells were treated with 2-48 µg/mL Nasturtium officinale gold nanoparticles (NOAuNPs) for 24 and 48 h to determine the effects on cell viability. The toxicity of the synthesized NOAuNPs to lung cancer cells was determined by the 3-(4,5-dimethylthiazol-2-il)-2,5-diphenyltetrazolium bromide (MTT) assay, and the anticancer effect of the NOAuNPs was evaluated by apoptosis and cell cycle analyses using flow cytometry. RESULTS: The average size of the NPs was 56.4 nm. The intensities of the Au peaks from EDS analysis indicated that the AuNPs were synthesized successfully. Moreover, the in vitro antioxidant activities of the NO and NOAuNPs were evaluated; these materials gave values of 31.78 ± 1.71% and 31.62 ± 0.46%, respectively, in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay at 200 g/mL and values of 25.89 ± 1.90% and 33.81 ± 0.62%, respectively, in the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay. The NO and NOAuNPs gave values of 0.389 ± 0.027 and 0.308 ± 0.005, respectively, in the ferrous ion reducing antioxidant capacity (FRAP) assay and values of 0.078 ± 0.009 and 0.172 ± 0.027, respectively, in the copper ion reducing antioxidant capacity (CUPRAC) assay. When the DNA cleavage activities of NO and the NOAuNPs were evaluated via hydrolysis, both samples cleaved DNA starting at a concentration of 25 g/mL in the cell culture analysis, while the nanoformulation of the NO components gave greater therapeutic and anticancer effects. We determined that the Au nanoparticles were not toxic to A549 cells. Moreover, after treatment with the half-maximal inhibitory concentration (IC50), determined by the MTT assay with A549 cells, we found that at 24 and 48 h, while the necrosis rates were high in cells treated with NO, the rates of apoptosis were greater in cells treated with NOAuNPs. Notably, for anticancer treatment, activating apoptotic pathways that do not cause inflammation is preferred. We believe that these results will pave the way for the use of NOAuNPs in in vitro studies of other types of cancer. CONCLUSION: In this study, AuNPs were successfully synthesized from N. officinale extract. The biosynthesized AuNPs exhibited toxicity to and apoptotic effects on A549 lung cancer cells. Based on these findings, we suggest that green-synthesized AuNPs are promising new therapeutic agents for lung cancer treatment. However, since this was an in vitro study, further research should be performed in in vivo lung cancer models to support our findings and to explain the mechanism of action at the molecular level.

Effects of Drying Methods on the Phytochemical Contents, Antioxidant Properties, and Anti-Diabetic Activity of Nasturtium officinale R.Br. (Betong Watercress) from Southern Thailand.

Nasturtium officinale R.Br. (Betong watercress) contains high levels of secondary metabolites that offer health benefits. However, fresh-cut watercress has a short shelf life. This study aimed to assess the effect of drying methods on the phytochemical contents, antioxidant activity, and anti-diabetic activity of Betong watercress. The watercress was dried using three methods: roasting (R, 50 °C, 40 min); roasting and drying (RD, 40 min roasting at 50 °C and 1 h drying at 80 °C); and blanching, roasting, and drying (BRD, 30 s blanching at 80 °C, 20 min roasting at 50 °C, and 1 h drying at 80 °C). Aqueous extracts from each drying method were analyzed for total phenolic content, total flavonoid content, total glucosinolate content, antioxidant activities (FRAP, DPPH, and ABTS assays), and α-amylase enzyme inhibition. From the results, the R method provided the highest level of total phenolic, total flavonoid, and total glucosionolate content compared to the RD and BRD methods. Similarly, antioxidant activities and α-amylase enzyme inhibition were highest in the R method, followed by the RD and BRD methods. Our results demonstrate that roasting of Betong watercress without the addition of blanching or drying effectively preserves the phytochemical contents, antioxidant activities, and anti-diabetic activity.

Making watercress (Nasturtium officinale) cropping sustainable: genomic insights into enhanced phosphorus use efficiency in an aquatic crop.

Watercress (Nasturtium officinale) is a nutrient-dense salad crop with high antioxidant capacity and glucosinolate concentration and with the potential to contribute to nutrient security as a locally grown outdoor aquatic crop in northern temperate climates. However, phosphate-based fertilizers used to support plant growth contribute to the eutrophication of aquatic habitats, often pristine chalk streams, downstream of farms, increasing pressure to minimize fertilizer use and develop a more phosphorus-use efficient (PUE) crop. Here, we grew genetically distinct watercress lines selected from a bi-parental mapping population on a commercial watercress farm either without additional phosphorus (P-) or under a commercial phosphate-based fertilizer regime (P+), to decipher effects on morphology, nutritional profile, and the transcriptome. Watercress plants sustained shoot yield in P- conditions, through enhanced root biomass, but with shorter stems and smaller leaves. Glucosinolate concentration was not affected by P- conditions, but both antioxidant capacity and the concentration of sugars and starch in shoot tissue were enhanced. We identified two watercress breeding lines, with contrasting strategies for enhanced PUE: line 60, with highly plastic root systems and increased root growth in P-, and line 102, maintaining high yield irrespective of P supply, but less plastic. RNA-seq analysis revealed a suite of genes involved in cell membrane remodeling, root development, suberization, and phosphate transport as potential future breeding targets for enhanced PUE. We identified watercress gene targets for enhanced PUE for future biotechnological and breeding approaches enabling less fertilizer inputs and reduced environmental damage from watercress cultivation.

Environmental and anthropogenic drivers of watercress (Nasturtium officinale) communities in char-lands and water channels across the Swat River Basin: implication for conservation planning.

Recent anthropogenic sources and excess usage have immensely threatened the communities and habitat ecology of this region's medicinally and economically significant crops. Therefore, our study aims to evaluate the community structure and related environmental characteristics sustaining Nasturtium officinale communities along the river basin (RB) in Northwest Pakistan, using the clustering procedure (Ward's method) and Redundancy analysis (RDA). From 340 phytosociological plots (34 × 10 = 340), we identified four ecologically distinct assemblages of N. officinale governed by different environmental and anthropogenic factors for the first time. The floristic structure shows the dominance of herbaceous (100%), native (77%), and annual (58.09%) species indicating relatively stable communities; however, the existence of the invasive plants (14%) is perturbing and may cause instability in the future, resulting in the replacement of herbaceous plant species. Likewise, we noticed apparent variations in the environmental factors, i.e., clay percentage (p = 3.1 × 10-5), silt and sand percentage (p< 0.05), organic matter (p< 0.001), phosphorus and potassium (p< 0.05), and heavy metals, i.e., Pb, Zn, and Cd (p< 0.05), indicating their dynamic role in maintaining the structure and composition of these ecologically distinct communities. RDA has also demonstrated the fundamental role of these factors in species-environment correlations and explained the geospatial variability and plants' ecological amplitudes in the Swat River wetland ecosystem. We concluded from this study that N. officinale communities are relatively stable due to their rapid colonization; however, most recent high anthropogenic interventions especially overharvesting and sand mining activities, apart from natural enemies, water deficit, mega-droughts, and recent flood intensification due to climate change scenario, are robust future threats to these communities. Our research highlights the dire need for the sustainable uses and conservation of these critical communities for aesthetics, as food for aquatic macrobiota and humans, enhancing water quality, breeding habitat, fodder crop, and its most promising medicinal properties in the region.

First identification of Xanthomonas nasturtii as the cause of black rot of watercress in Hawaii.

Watercress (Nasturtium officinale) has been in continuous production in Hawaii for over a century and is part of the local diet. Black rot of watercress was first identified as caused by Xanthomonas nasturtii in Florida (Vicente et al., 2017), but symptoms of this disease have also been regularly observed in Hawaii production in all islands, mostly during the rainy season from December to April in areas with poor air circulation (McHugh & Constantinides, 2004). Initially, this disease was attributed to X. campestris due to similar symptoms to black rot of brassicas. Samples of watercress with symptoms that could be attributed to a bacterial disease including yellow spots and lesions on leaves and stunting and deformation of plants in more advanced stages, were collected from a farm in Aiea in the island of Oahu, Hawaii, in October 2017. Isolations were performed at the University of Warwick. Fluid from macerated leaves was streaked into plates of King's B (KB) medium and Yeast Dextrose Calcium Carbonate Agar (YDC). After 48-72 hrs incubation at 28°C, the plates showed a range of mixed colonies. Single cream-yellow mucoid colonies were sub-cultured several times and pure isolates including WHRI 8984 were stored at -76°C as previously described (Vicente et al., 2017). Colony morphology was observed in KB plates and, in contrast to the type strain from Florida (WHRI 8853 = NCPPB 4600), isolate WHRI 8984 did not cause browning of the medium. Pathogenicity was tested on four-week old watercress and Savoy cabbage cv. Wirosa F1 plants by inoculations on leaves as previously described (Vicente et al., 2017). WHRI 8984 did not produce symptoms when inoculated on cabbage but produced typical symptoms on watercress. A re-isolation from a leaf showing a V-shaped lesion, produced isolates with the same morphology, including isolate WHRI 10007A, that was also shown to be pathogenic to watercress therefore completing the Koch's postulates. Fatty acid profiling was performed on WHRI 8984 and 10007A and controls grown on trypticase soy broth agar (TSBA) plates at 28°C for 48 hrs as described by Weller et al. (2000). Profiles were compared with the RTSBA6 v6.21 library; as the database does not include X. nasturtii, the results were only interpreted at the genus level, and both isolates were shown to be Xanthomonas sp. For molecular analysis, DNA was extracted and the gyrB partial gene was amplified and sequenced as described by Parkinson et al. (2007). Comparisons with sequences available in the National Centre for Biotechnology Information (NCBI) databases using the Basic Local Alignment Search Tool (BLAST) showed that partial gyrB of WHRI 8984 and 10007A were identical to the type strain from Florida therefore confirming that they belong to X. nasturtii. For whole genome sequencing, genomic libraries for WHRI 8984 were prepared using Illumina's Nextera XT v2 kit and sequenced on a HiSeq Rapid Run flowcell. The sequences were processed as previously described (Vicente et al., 2017) and the whole genome assembly has been deposited in GenBank (accession QUZM00000000.1); the phylogenetic tree shows that WHRI 8984 is close, but not identical to the type strain. This is the first identification of X. nasturtii in watercress crops in Hawaii. Control of this disease generally involves the use of copper bactericides and minimizing moisture on leaves by reducing overhead irrigation and increasing air circulation (McHugh & Constantinides, 2004); seed testing might help to select batches that are disease free and, in longer term, breeding for disease resistance might produce cultivars that can be part of management strategies.

GC-MS and HPLC supported phytochemical analysis of watercress and the protective role against paraben toxicity.

In this study, the phytochemical content of Nasturtium officinale R. Br. (watercress) leaf extract (Noex) and its protective effects against paraben toxicity were investigated. GC-MS and HPLC analyses were performed to determine the phytochemical content. Paraben toxicity and protective properties of Noex were investigated with the Allium test, and 6 different groups were formed for this purpose. Toxicity in each group was investigated by using physiological, cytogenetic, biochemical, and anatomical parameters. DNA-paraben interaction was investigated with spectroscopic analysis for the genotoxicity mechanism. As a result of the study, paraben (500 mM) caused a regression in the physiological parameters related to germination in Allium cepa L. bulbs. Paraben caused a 43.3% reduction in mitotic index (MI) rates compared to control, which is likely the reason for the decrease in germination-related parameters. With the application of paraben in root tip cells, the frequency of micronucleus (MN) and chromosomal aberrations (CAs) increased and a high genotoxic effect was observed. Paraben promoted CAs such as fragment, sticky chromosome, bridge, unequal distribution of chromatin, and irregular mitosis. It also caused anatomical damage in the form of epidermis cell damage, flattened cell nucleus, cortex cell damage, cortex cell walls thickening, and unclear vascular tissue in root tip meristem cells. Paraben-DNA interaction was caused by bathochromic and hypochromic shifts in the UV spectrum of DNA, indicating the intercalation mode of interaction. Paraben also caused an increase in malondialdehyde (MDA) levels, a decrease in glutathione (GSH) levels, and abnormalities in antioxidant enzyme levels (superoxide dismutase = SOD and catalase = CAT), thereby disrupting the antioxidant/oxidant dynamics in the cell. The basis of physiological, cytological, and genetic abnormalities was attributed to the oxidative stress in the cell. Administration of Noex produced a dose-dependent incremental improvement in paraben-induced abnormalities. The increase in GSH levels and the decrease in MDA levels observed as a result of the Noex application contributed to the restoration of antioxidant/oxidant balance, and this improvement was also reflected in other parameters. Application of 200 mg/L Noex provided a 24.2% improvement in the MI rate reduced by paraben, and accordingly, an increase in germination parameters was observed. Similarly, the frequencies of MN and CAs, which are signs of genotoxicity, decreased with the Noex application. As a result of the phytochemical analysis of Noex with HPLC and GC-MS, the presence of strong antioxidant and antimutagenic substances such as rutin, coumaric acid, ferrulic acid, L-serine, L-proline, and phytol were determined in Noex structure. The curative effects of Noex against paraben toxicity can be attributed to these active ingredients.

Salicylic acid improves Nasturtium officinale phytoremediation capability for cadmium-contaminated paddy soils.

Introduction: Cadmium (Cd) contamination is a severe problem in paddy soils that has affected crops' safety. The present study aimed at remediating Cd-contaminated paddy soil by improving the phytoremediation capability of aquatic accumulator plants. Methods: We conducted an experiment to investigate the effects of salicylic acid (SA) on the growth and Cd phytoremediation capability of the aquatic accumulator plant Nasturtium officinale. Results: SA with the concentrations of 100, 150, and 200 mg/L increased the root and shoot biomass of N. officinale, while only 150 mg/L increased the chlorophyll a and b contents. SA increased the activities of peroxidase and catalase of N. officinale to a great extent, but decreased the superoxide dismutase activity and soluble protein content. SA also increased the root Cd content, shoot Cd content, root Cd extraction, and shoot Cd extraction to a large extent. At concentrations of 100, 150, and 200 mg/L, SA increased the shoot Cd extraction by 17.59%, 47.16%, and 43.27%, respectively, compared with the control. Moreover, SA concentration had a quadratic polynomial regression relationship with the root Cd extraction and shoot Cd extraction. The correlation and grey relational analyses revealed that root Cd extraction, shoot biomass, and root biomass were closely associated with shoot Cd extraction of N. officinale. Conclusion: Thus, our results suggest that SA promoted the growth and improved the phytoremediation (extraction) capability of N. officinale, and 150 mg/L SA was the most suitable concentration.

Nasturtium officinale L. and metformin alleviate the estradiol- induced polycystic ovary syndrome with synergistic effects through modulation of Bax/Bcl-2/p53/caspase-3 signaling pathway and anti-inflammatory and anti-oxidative effects.

Polycystic ovary syndrome (PCOS) is one of the most common causes of infertility in women, which is associated with metabolic, hereditary and hormonal disorders. The aim of this study was to evaluate the therapeutic effects of Nasturtium officinale L. (N. officinale) on biochemical and molecular parameters in estradiol-induced PCOS in rats. Seventy Wistar rats in 7 groups (n = 10) were randomly assigned to normal (NC), PCOS, metformin (MET - 300 mg/kg), N. officinale (50 and 100 mg/kg) and co-treatment with MET and N. officinale groups. After 21 days of treatment, biochemical parameters levels of estrogen, LH and FSH along with serum levels of (IL-6 and IL-1ß cytokines) and serum antioxidant parameters (enzymatic activity of catalase and superoxide dismutase) were measured. Finally, by measuring the expression of apoptosis related genes (Bax/Bcl-2/p53/caspase-3) with the help of real-time PCR and the expression of p53 with the help of immunohistochemistry in ovarian cells. N. officinale modulates hormones through its hypothalamic-pituitary-gonadal pathway with its synergistic effects along with MET. Also, in co-treatment groups (MET and N. officinale), the activity of serum antioxidant enzymes increased and also the serum level of inflammatory cytokines decreased. N. officinale, along with MET, amplified the Bax/Bcl2/p53/caspase-3 pathways, which eventually increased the number of p53 positive cells. These findings indicate that N. officinale extract along with MET can improve the physiological function of the ovaries in PCOS-induced disorders. PRACTICAL APPLICATIONS: Polycystic ovary syndrome (PCOS) is one of the most common causes of infertility in women, which is associated with metabolic, hereditary and hormonal disorders. The extract of Nasturtium officinale L. was able to intensify mitochondrial apoptotic pathway in cystic follicles and prevent their formation. It seems that pro-drugs containing N. officinale along with effective commercial drugs in PCOS can help ovulation and fertility in woman with this disease.

Cloning and Functional Analysis of NoMYB60 Gene Involved in Flavonoid Biosynthesis in Watercress (Nasturtium officinale R. Br.).

The MYB60 gene belongs to the R2R3-MYB subfamily, which includes the MYB31/30/96/94 genes. Although these genes have been shown to respond to heat and drought stresses, their role in flavonoid synthesis remains unclear. In this study, NoMYB60 was cloned from watercress and its structure and function were analyzed. Sequence structure analysis showed that NoMYB60 had a highly conserved R2R3 DNA-binding region at the N-terminus. Under the treatment of ABA, SA or MeJA, the expression level of NoMYB60 first significantly increased and then decreased, indicating that ABA, SA and MeJA positively regulated NoMYB60. The subcellular localization of NoMYB60-GFP indicated that NoMYB60 was localized in the nuclear region, which is consistent with the molecular characterization of the transcription factor. Gene silencing experiments were also performed to further test the function of NoMYB60. The result showed that virus-induced silencing of NoMYB60 affected the expression of enzyme genes in flavonoid synthesis pathways and promoted the synthesis of flavonoids. Moreover, we discovered that NoMYB60 interacts with NoBEH1/2. In this study, provides a reference for research on the regulation mechanism of flavonoid synthesis in Cruciferae and other crops.

An amino acid fertilizer improves the emergent accumulator plant Nasturtium officinale R. Br. phytoremediation capability for cadmium-contaminated paddy soils.

Cadmium (Cd) contamination of paddy soil affects safe crop production. This study aimed to evaluate the effects of plant biostimulant amino acid fertilizer on the phytoremediation capability of an emergent accumulator plant Nasturtium officinale R. Br. for Cd-contaminated paddy soils. A pot study was carried out to study the effects of different concentrations of amino acid fertilizer on the Cd accumulation of N. officinale grown in Cd-contaminated paddy soil. The amino acid fertilizer increased the biomass of N. officinale. The amino acid fertilizer concentration exhibited a quadratic polynomial regression relationship with the root and shoot biomass. The fertilizer also increased the photosynthetic pigment (chlorophyll and carotenoid) contents, peroxidase (POD; EC 1.11.1.7) activity, and catalase (CAT; EC 1.11.1.6) activity of N. officinale, but decreased the soluble protein content and had no significant effect on the superoxide dismutase (SOD; EC 1.15.1.1) activity. Furthermore, the amino acid fertilizer increased the Cd content and Cd extraction of N. officinale. The shoot Cd extraction increased by 29.06%, 63.05%, 77.22%, and 17.40% at 1500-, 1200-, 900-, and 600-fold dilutions of the amino acid fertilizer, respectively, compared with the control. Moreover, the amino acid fertilizer promoted the Cd transport from the roots to shoots of N. officinale. The amino acid fertilizer concentration also exhibited a quadratic polynomial regression relationship with the root Cd content, shoot Cd content, root Cd extraction, and shoot Cd extraction, respectively. The correlation, grey relational, and path analyses revealed that the root biomass, shoot biomass, chlorophyll content, catalase activity, shoot Cd content, and root Cd extraction were closely associated with the shoot Cd extraction. Therefore, the amino acid fertilizer can promote Cd uptake and improve the phytoremediation capability of N. officinale to remediate Cd-contaminated paddy soils, and 900-fold dilution is the most suitable concentration.

The antibacterial activity of nasturtium officinale extract on common oral pathogenic bacteria.

Background: The oral cavity is colonized by a myriad of microorganisms, some of which are proven to be detrimental to human health. There have been numerous efforts to control the population of pathogenic agents in the oral cavity, including the usage of natural phytochemicals obtained from medicinal plants. Nasturtium officinale has long been used in traditional medicine for the management of hypertension, respiratory infections, and hyperglycemia, and its effectiveness against some microbes has been reported. Aims: To evaluate antimicrobial properties of a hydro-alcoholic extract of N. officinale against common oral pathogens namely Streptococcus mutans, Staphylococcus aureus, Lactobacillus acidophilus, Enterococcus faecalis, and Pseudomonas aeruginosa. Experimental laboratory study. Different dilutions of N. officinale hydro-alcoholic extract were the test solutions, the positive control was a bacterial suspension in sterile phosphate-buffered saline, whereas the negative control was the herbal extract only, without any bacterial inoculation. Hydro-alcoholic extract of N. officinale prepared in five different concentrations (105, 52.5, 26.25, 13.12, 6.56 mg.mL-1) was tested separately against Streptococcus mutans, Lactobacillus acidophilus, Pseudomonas aeruginosa, Enterococcus faecalis, and Staphylococcus aureus in a test of microdilution assay. Spectrophotometry was used to assess bacterial growth after 24 and 48 h. Materials and Methods: The data of optical absorbance reads from spectrophotometry were analyzed using repeated-measures analysis followed by Least Significant Differences (LSD) post hoc. Results: The highest growth inhibitory effect against S. mutans, E. faecalis, and S. aureus was observed at a concentration of 13.12 mg.mL-1; for L. acidophilus and P. aeruginosa, the most significant inhibition was observed at a concentration of 105 mg.mL-1. Conclusion: N. officinale extract effectively inhibited the growth of the tested oral bacteria at different concentrations but was more effective against S. mutans, E. faecalis, and S. aureus and so may be effective in managing some oral microbial infections.

Further evidence to support acute and chronic anti-inflammatory effects of Nasturtium officinale.

Background and purpose: Previously, we reported the anti-inflammatory properties of Nasturtium officinale (watercress) in several models of acute inflammation. This study was designed to explore the effects of topical and systemic administrations of N. officinale in the two chronic inflammatory models and to evaluate the role of TNF-α and IL-1ß in these effects. Experimental approach: Folin-Ciocalteu and aluminum chloride methods were used to estimate the extract's total phenol and flavonoid content, respectively. Carrageenan-induced paw edema was carried out and TNF-α and IL-1ß concentrations in the carrageenan-treated paw tissue were determined. Formalin injection into rat hind paws (7 days) and the application of 12-O-tetradecanoyl phorbol-13-acetate (TPA) on mouse ears (9 days) were used to simulate chronic inflammation. Furthermore, a histological assessment of the inflamed tissues was carried out. Findings/Results: The extract's flavonoid and phenolic contents were 90.26 ± 4.81 mg rutin equivalents/g and 68 ± 8.16 gallic acid equivalents/g gallic acid, respectively. N. officinale pretreatment in all doses administered considerably decreased carrageenan-induced edema. The extract also reduced IL-1ß levels in carrageenan- treated paws while did not affect TNF-α levels. Oral and topical administrations of N. officinale considerably reserved the paw and ear edema. The extract also ameliorated the tissue injuries due to formalin and TPA challenges. Conclusion and implications: The data confirmed the topical and systemic anti-inflammatory effects of watercress against two chronic models of inflammation. They suggested that these properties are not related to TNF-α but could be attributed to inhibition of IL-1ß and inhibition of leukocyte infiltration.

Efficacy of synergistic activity of seed oils from Carthamus tinctorius (Safflower) and Nasturtium officinale (Watercress) on the lethality of the cattle tick Hyalomma scupense (Acari: Ixodidae).

Background: Ticks and tick-borne diseases are a severe economic and public-health problem for cattle producers. The emergence of acaricide resistance to synthetic chemical treatments has prompted interest in developing alternative tick control methods. Aim: The main objective of the current research was to identify the chemical structure of Carthamus tinctorius and Nasturtium officinale seed oils and to assess their anti-tick properties against Hyalomma scupense ticks both alone and in combination (1:1). Methods: Analytical methods were used to analyze the chemical components. For in vitro assays, adults of H. scupense were immersed in C. tinctorius and N. officinale seed oils at 100, 125, 200, and 300 mg/ml concentrations; for 5 minutes. Larvae of H. scupense were dipped in 25, 50, 100, 125, 200, and 300 mg/ml doses of seed oils; the mortality percentage was determined after 24 hours. Results: The seed oil safflower was mainly composed of linoleic acid (84.48%), followed by palmitic acid (6.54%) and stearic acid (3.77%). Meanwhile, watercress seed oil was mainly composed of linolenic acid (50.78%), gondoic acid (13.57%), linoleic acid (10.58%), palmitic acid (8.02%), and erucidic acid (6.62%). The Adults Immersion Test showed the sensitivity of ticks to C. tinctorius and N. officinale seed oil: C. tinctorius seed oil caused (95%) mortality of H. scupense at 300 mg/ml, while N. officinale seed oil induced (88.68%) mortality at the same concentration. At a 200 mg/ml concentration, C. tinctorius and N. officinale oil combined caused 100% mortality. Tested oils showed larvicidal efficacy. LC50 values for C. tinctorius and N. officinale seed oils were 84.16 and 61.78 mg/ml, respectively, in 24 hours. LC50 value of oils association (50% C. tinctorius: 50% N. officinale) was 47.96 mg/ml.The mixture of seed oils from two plants tested against H. scupense larvae and adult females at a 1:1 ratio showed synergistic interaction. Conclusion: Seed oils tested alone, and the mixture could be used as an alternative solution in the fight against ticks.

Impacts of elicitors on metabolite production and on antioxidant potential and tyrosinase inhibition in watercress microshoot cultures.

The study has proved the stimulating effects of different strategies of treatments with elicitors on the production of glucosinolates (GSLs), flavonoids, polyphenols, saccharides, and photosynthetic pigments in watercress (Nasturtium officinale) microshoot cultures. The study also assessed antioxidant and anti-melanin activities. The following elicitors were tested: ethephon (ETH), methyl jasmonate (MeJA), sodium salicylate (NaSA), and yeast extract (YeE) and were added on day 10 of the growth period. Cultures not treated with the elicitor were used as control. The total GSL content estimations and UHPLC-DAD-MS/MS analyses showed that elicitation influenced the qualitative and quantitative profiles of GSLs. MeJA stimulated the production of gluconasturtiin (68.34 mg/100 g dried weight (DW)) and glucobrassicin (65.95 mg/100 g DW). The elicitation also increased flavonoid accumulation (max. 1131.33 mg/100 g DW, for 100 µM NaSA, collection after 24 h). The elicitors did not boost the total polyphenol content. NaSA at 100 µM increased the production of total chlorophyll a and b (5.7 times after 24 h of treatment), and 50 µM NaSA caused a 6.5 times higher production of carotenoids after 8 days of treatment. The antioxidant potential (assessed with the CUPRAC FRAP and DPPH assays) increased most after 24 h of treatment with 100 µM MeJA. The assessment of anti-melanin activities showed that the microshoot extracts were able to cause inhibition of tyrosinase (max. 27.84% for 1250 µg/mL). KEY POINTS: • Elicitation stimulated of the metabolite production in N. officinale microshoots. • High production of pro-health glucosinolates and polyphenols was obtained. • N. officinale microshoots have got tyrosinase inhibition potential. • The antioxidant potential of N. officinale microshoots was evaluated.

Manipulation of Continuous and End-of-Day Red/Far-Red Light Ratios Affects Glucobrassicin and Gluconasturtiin Accumulation in Cabbage (Brassica oleracea) and Watercress (Nasturtium officinale), Respectively.

Cabbage (Brassica oleracea) and watercress (Nasturtium officinale) produce glucobrassicin (GBS) and gluconasturtiin (GNST), precursors of chemopreventive compounds. Their accumulation is affected by environmental signals. We studied the impact of the red to far-red light (R/FR) ratio on GBS concentration in red ″Ruby Ball″ and green ″Tiara″ cabbage. Foliar shading, via weed surrogates that competed with cabbage plants for specific durations, induced R/FR variation among treatments. ″Ruby Ball″ GBS concentrations were the highest when R/FR within the canopy was the lowest. ″Tiara″ was unaffected by competition. The same trend was observed in a controlled environment using R and FR LEDs without weeds present. ″Ruby Ball″ subjected to an R/FR = 0.3 treatment had 2.5- and 1.4-fold greater GBS concentration compared to R/FR = 1.1 and 5.0 treatments combined. Watercress given end-of-day (EOD) R and/or FR pulses after the main photoperiod had the lowest GNST concentrations after an EOD FR pulse but the highest concentrations after an R followed by FR pulse.

Analysis of Flavonoid Metabolites in Watercress (Nasturtium officinale R. Br.) and the Non-Heading Chinese Cabbage (Brassica rapa ssp. chinensis cv. Aijiaohuang) Using UHPLC-ESI-MS/MS.

Flavonoids from plants play an important role in our diet. Watercress is a special plant that is rich in flavonoids. In this study, four important watercress varieties were compared with non-heading Chinese cabbage by ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS). A total of 132 flavonoid metabolites (including 8 anthocyanins, 2 dihydroflavone, 3 dihydroflavonol, 1 flavanols, 22 flavones, 11 flavonoid carbonosides, 82 flavonols, and 3 isoflavones) were detected. Flavonoid metabolites varied widely in different samples. Both the non-heading Chinese cabbage and the variety of watercress from Guangdong, China, had their own unique metabolites. This work is helpful to better understand flavonoid metabolites between the non-heading Chinese cabbage and the other four watercress varieties, and to provide a reliable reference value for further research.

First report of Sclerotinia sclerotiorum on watercress (Nasturtium officinale) in aquaponic system in Hungary.

Watercress (Nasturtium officinale) is an aquatic dicotyledonous vegetable belonging to Brassicaceae (Aiton 1812). Watercress was grown in an aquaponic system on fired clay ball medium at the Aquaponic Research Station of the University of Debrecen, in the city of Debrecen (Hungary). During January 2020, 3-month-old plants showed symptoms in aquaponic cultivation. A visual survey showed 30% of plants with symptoms. Leaves and stems withered and showed white cotton-like mycelium. Mycelia from infected plants were placed on potato dextrose agar (PDA) and incubated at 25°C for seven days. Single hyphal tips were transferred to produce a pure culture. All ten fungal isolates showed similar morphological characteristics on PDA. Colonies consisted of white mycelia after three days and globoid to irregular and black 2.5 to 7 (average, 3) mm (n = 100 from ten plates) sclerotia formed ten days later, which are the typical morphological features of Sclerotinia sclerotiorum (Mordue et al. 1976). Molecular identification was performed with one of the ten isolates (Scl_B). Mycelia were grown in 250 ml of potato dextrose broth in a rotary shaker at 175 rpm at 24°C for six days. DNA was extracted from mycelium using a Nucleospin plant II (Macherey-Nagel, Germany) according to the manufacturer's protocol. PCR amplification (Kim et al. 2014) was performed with primers ITS1/ITS4 for the internal transcribed spacer region (White et al. 1990) on a Primus 96 thermal cycler (MWG Biotech, Germany). Specific polymerase chain reaction was performed with primers SSasprF/SSasprR (Abd-Elmagid et al. 2013). PCR products were sequenced by Microsynth Austria GmbH. NCBI BLAST analysis of the 440-bp ITS sequence (Genbank MW012403.1) showed 100% identity with the sequence of S. sclerotiorum (MT177267.1, etc.). The 170-bp specific gene sequence (Genbank MW959042.1) had a 100% similarity to hypothetical proteins (Genbank MK028159.1), with a 99.4% similarity to a portion of the S. sclerotiorum aspartyl protease gene (AF271387.1). Pathogenicity tests were carried out by inoculating surface-disinfested, 30-day-old watercress plants in plastic pots (15x15x12 cm). In three repeated experiments 90 watercress plants were measured. 15 plants (one plant per pot) were planted into the five-times autoclaved substrate (Biorgmix: pH 6.1±0.5%, N:1.5%, P2O5:0.7%, K2O:0.5%, organic matter content:50%) and inoculated by ten wheat kernels that were colonized by S. sclerotiorum (Scl_B) (Garibaldi et al. 2019). 15 plants were planted into the substrate with ten non-inoculated kernels as a control. Plants were kept in an MLR-352 climatic test chamber (PHCbi, Japan) at 21 ± 1°C for 12 hr light:dark cycle. On the first day of the experiment complex nutrient solution (Tek-Land: N:5%, P2O5:5%, K2O:5%, B:0.01%, Cu:0,01%, Mn:0.02%, Mo:0.002%, Zn:0.016%) was used, then autoclaved water daily. Eight days later white mycelium appeared on every inoculated plant and five days later dark sclerotia formed on the stems. Based on the morphological characteristics the re-isolated pathogen was S. sclerotiorum. Similar results were detected in three repeated experiments with white mold fungus being reisolated from all 45 infected watercress plants. The 45 non-inoculated plants did not show any symptoms and any diseases. This pathogen has already been reported on watercress in the field (Farr et al. 1989; Boland and Hall 1994; Garibaldi et al. 2019). This is the first reported case of white mold on watercress in aquaponic system in Hungary.

Protective and therapeutic effects of ethanolic extract of Nasturtium officinale (watercress) and vitamin E against bleomycin-induced pulmonary fibrosis in rats.

BACKGROUND AND PURPOSE: Pulmonary fibrosis is a chronic disease of the lungs caused by inflammation, species of reactive oxygen, and immune defects. Antioxidant properties of Nasturtium officinale has been reported in some studies. Therefore, the objective of the current study was to evaluate the effect of ethanolic extract of Nasturtium officinale (EENO) on bleomycin (BLM)-induced lung fibrosis in rats. EXPERIMENTAL APPROACH: Forty adult male Wistar rats (180-220 g) were randomly divided into 5 experimental groups. Normal control, BLM control received a single dose of BLM (6 IU/kg) intratracheally only on the first day, EENO + BLM group received EENO (500 mg/kg) one week before intratracheal BLM instillation and two weeks afterward, BLM + EENO group and BML + vitamin E group received EENO (500 mg/kg) and vitamin E (500 mg/kg) half-hour after BLM installation, respectively. The animals were sacrificed on day 22. Change in body weight, lung index, serum level of malondialdehyde (MDA) and nitric oxide (NO) metabolite, lung tissue hydroxyproline content and lung pathology were assessed. FINDINGS/RESULTS: Pre- or post-treatment with EENO attenuated pulmonary fibrosis as evidenced by normalized lung index, improved histological changes and inhibited collagen deposition (hydroxyproline) in the animal lung. EENO also decreased MDA and NO metabolite release in comparison to the BLM control. vitamin E (500 mg/ kg) also significantly inhibited the BLM-induced lung toxicity. CONCLUSIONS AND IMPLICATIONS: EENO can prevent BLM-induced lung fibrosis in rats via antioxidant activities. However, more studies are needed to elicit the exact mechanism of this effect.

Effect of watercress extract supplementation on lipid profile and oxidative stress markers in overweight people with physical disability: A randomized, double-blind, and placebo-controlled trial.

Studies have demonstrated that diet rich in cruciferous vegetables of the Brassicaceae family can reduce the risk of cardiovascular diseases and oxidative stress levels. Nasturtium officinale (Brassicaceae), commonly known as watercress is a perennial dicotyledonous plant usually found close to water. Although previous investigations have demonstrated the beneficial effects of watercress on hypercholesterolemia in animal studies, until now no such studies have been conducted with humans, up to this time. This study aimed to investigate whether overweight individuals were able to improve or maintain their serum lipid and oxidative stress markers when given standardized extract of Nasturtium officinale (SENO) as a supplement. This was a randomized, double-blind, and placebo-controlled trial conducted over 5 weeks. Thirty-four overweight people with physical disabilities were selected randomly to participate in this study and then they were assigned randomly to two groups, one treated with 750 mg//kg/d of SENO and the other treated with 750 mg/kg/d of placebo. The results indicated that SENO caused a significant improvement in the levels of low-density lipoprotein cholesterol, creatinine, and lipid peroxidation. However, SENO did not cause a significant statistical change in total serum cholesterol, triacylglycerol, and high-density lipoprotein levels; catalase, superoxide dismutase, creatinine, alanine aminotransferase, aspartate aminotransferase, and urea parameters. The present data might provide supportive evidence that SENO did not cause any harm and positively affected low-density lipoprotein cholesterol profile and creatinine as well as lipid peroxidation levels in the participants. Nevertheless, further studies are suggested to clarify the results presented in this clinical trial.

Biomarkers of oxidative stress and inflammation in people witha physical disability treated with a standardized extract of Nasturtium officinale: A randomized, double-blind, and placebo-controlled trial.

It is well established that plants from the Brassicaceae family, particularly watercress, have been associated to reduce oxidative DNA damage. Nasturtium officinale R. Br (watercress) contains glucosinolates, with anti-inflammatory action and protective effect on human health against oxidative stress. We aimed to evaluate whether the standardized extract of Nasturtium officinale (SENO) is capable of changing biomarkers of oxidative stress and inflammation in people with physical disabilities. 65 people enrolled this study: as a control group composed by; 15 people with no physical disability assessed once, 25 people with physical disabilities using 750 mg/kg/day of SENO, and 25 people with physical disabilities using 750 mg/kg/day of placebo-control for 5 weeks. Biomarkers of oxidative stress and inflammation were analyzed on day 0 and 36. The results indicated that SENO was associated with decreasing levels of lipid peroxidation, protein carbonyl, catalase, superoxide dismutase, and C-reactive protein. Furthermore, the cytokine kit demonstrated below and out of invertible range, which was impossible to detect the inflammatory process. Despite the cytokine kit was not able to detect the inflammation; these data might provide supportive evidence that SENO, have affected positively people with physical disabilities decreasing their biomarkers of oxidative stress and C-reactive protein. Further studies are required.