Investigation of combined effects of propyl paraben and methyl paraben on the hypothalamic-pituitary-adrenal axis in male rats.

The aim of this study was to investigate the endocrine-disrupting effects of methyl paraben (MeP) and propyl paraben (PrP) mixture on the hypothalamic-pituitary-adrenal axis (HPA). In this study, six experimental groups were designated. These groups included three control groups (control, corn oil control, and positive control (50 mg/kg/day BPA)) and three dose groups (10, 100, and 500 mg/kg/day MeP+PrP). MeP with PrP were mixed in a 1:1 ratio and administered to the 42-day-old male rats by oral gavage for 30 days. At the end of the experiment, adrenocorticotropic hormone (ACTH), corticosterone and aldosterone hormones were analyzed in serum. Effects of MeP+PrP on the adrenal glands were investigated by immunohistochemical staining of 11ß hydroxylase (CYP11B1) and aldosterone synthase (CYP11B2) enzymes involved in the synthesis steps of corticosterone and aldosterone. Also, pituitary and adrenal glands were examined histopathologically. In the histopathological findings, cortical nodule, congestion, and edema were found in the tissues. In the pituitary gland, cytokeratin rings were detected in all MeP+PrP dose groups, supporting the increase of corticosterone and ACTH. Serum corticosterone, aldosterone, and ACTH hormone levels were increased in the 100 mg/kg/day MeP+PrP and BPA groups. Results obtained from immunohistochemical staining showed that increased staining parallelled increased corticosterone and aldosterone hormone levels. In summary, the results showed that exposure to the MeP+PrP mixture caused a significant increase in ACTH and corticosterone. Also, the MeP+PrP mixture caused a significant increase of CYP11B1 and CYP11B2. MeP+PrP exposure disrupts the normal HPA axis.

A Novel In Vitro Assay Using Human iPSC-Derived Sensory Neurons to Evaluate the Effects of External Chemicals on Neuronal Morphology: Possible Implications in the Prediction of Abnormal Skin Sensation.

Neuronal morphological changes in the epidermis are considered to be one of causes of abnormal skin sensations in dry skin-based skin diseases. The present study aimed to develop an in vitro model optimised for human skin to test the external factors that lead to its exacerbation. Human-induced pluripotent stem cell-derived sensory neurons (hiPSC-SNs) were used as a model of human sensory neurons. The effects of chemical substances on these neurons were evaluated by observing the elongation of nerve fibers, incidence of blebs (bead-like swellings), and the expression of nicotinamide mononucleotide adenylyl transferase 2 (NMNAT2). The nerve fiber length increased upon exposure to two common cosmetic preservatives-methylparaben and phenoxyethanol-but not to benzo[a]pyrene, an air pollutant at the estimated concentrations in the epidermis. Furthermore, the incidence of blebs increased upon exposure to benzo[a]pyrene. However, there was a decrease in the expression of NMNAT2 in nerve fibers, suggesting degenerative changes. No such degeneration was found after methylparaben or phenoxyethanol at the estimated concentrations in the epidermis. These findings suggest that methylparaben and phenoxyethanol promote nerve elongation in hiPSC-SNs, whereas benzo[a]pyrene induces nerve degeneration. Such alterations may be at least partly involved in the onset and progression of sensitive skin.

Effects of Butylparaben Supplementation on In Vitro Development of Mouse Preantral Follicle.

We investigated whether butylparaben supplementation to the culture media negatively effects on in vitro development of mouse preantral follicle. The preantral follicles were isolated from the ovaries of 7-8-week-old mice and cultured in growth medium for 10 days and then in maturation medium for 2 days. During in vitro culture, butylparaben (0, 0.01, 0.1, 1.0, or 10 µM) was supplemented to the culture media. In the final spent media, the levels of 17ß-estradiol and anti-Müllerian hormone (AMH) were measured via enzyme-linked immunosorbent assay. In the final luteinized follicular cells, the mRNA levels of steroidogenic acute regulatory protein (StAR), superoxide dismutase 1 (Sod1), caspase 3 (Casp3), and extracellular signal-regulated kinase 1 (Erk1) were quantified via real-time reverse transcription-polymerase chain reaction. The metaphase II oocyte acquisition (per total oocyte) tended to decrease in the four butylparaben-supplemented groups, but not significant (26.8%, 23.2%, 21.4%, 15.1%, and 16.8%, respectively). The level of 17ß-estradiol and AMH tended to decrease in all butylparaben-supplemented groups, but statistically not significant. The expression level of StAR and Erk1 mRNA was significantly higher in all four butylparaben-supplemented groups, and a dose-dependent increment tendency was observed. Our findings suggest that butylparaben supplementation has largely no impact on in vitro development of mouse preantral follicle as well as 17ß-estradiol and AMH production. However, StAR, Sod1, Casp3, and Erk1 genes could be overexpressed in a certain concentration of butylparaben.

Combination of natural antimicrobials for contamination control in ethanol production.

Presence of bacterial contaminants at levels > 107 colony forming units per milliliter (CFU/mL) during ethanol production processes reduces the alcoholic fermentation yield by 30%. Antibiotics are currently used to control contamination, but their residues may be detected in yeast extract, restricting this by-product trade to several countries. Thus, the objective of this study was to assess antimicrobial activity of the natural compounds hops extract, 4-hydroxybenzoic acid, nisin Z, and lysozyme against Lactobacillus fermentum, Leuconostoc mesenteroides, and Saccharomyces cerevisiae, aiming development of a formula. Minimum Inhibitory Concentration of each antimicrobial was determined for bacteria and subsequently, nisin (30 mg/L) and hops extract (5 mg/L) were tested together, showing inhibitory effects combining doses of each antimicrobial that were equivalent to an eightfold reduction of their original Minimum Inhibitory Concentrations (3.75 and 0.625 mg/L, respectively), resulting in a FICIndex of 0.25. Thereon, a formula containing both compounds was developed and tested in fermentation assays, promoting reductions on bacterial population and no severe interferences in yeast viability or population even at extreme doses. Therefore, these compounds have great potential to successfully substitute conventional antibiotics in the ethanol industry.

Protein disulfide isomerases as potential therapeutic targets for influenza A and B viruses.

Seasonal flu as well as potential pandemic flu outbreaks continuously underscores the importance of the preventive and therapeutic measures against influenza viruses. During screening of natural and synthetic small molecules against influenza A and B virus, we identified juniferdin as a highly effective inhibitor against both viruses in cells. Since juniferdin is known to inhibit protein disulfide isomerases (PDIs), multiple PDI inhibitors were tested against these viruses. Among PDI inhibitors, 16F16, PACMA31, isoquercetin, epigallocatechin-3-gallate or nitazoxanide significantly reduced the replication of influenza A and B viruses in MDCK and A549 cells. Furthermore, siRNAs specific to three PDI family members (PDI1, PDIA3 or PDIA4) also significantly reduced the replication of influenza A and B viruses in cells. These results suggest that PDIs may serve as excellent targets for the development of new anti-influenza drugs.

Transformed Root Extract of Leonurus sibiricus Induces Apoptosis through Intrinsic and Extrinsic Pathways in Various Grades of Human Glioma Cells.

This study determines the influence of transformed root (TR) extract of Leonurus sibiricus L. on various grades (I-III) of human glioma cells derived from patients. This plant occurs in southern Asia and Siberia and is widely used as a medicinal plant with various biological activities. Chromatographic profile of TR extract have revealed the presence of various polyphenolic compounds (4-hydroxybenzoic acid, gentisic acid, vanilic acid, 1,3-dicaffeoylquinic acid, α-resorcylic acid). We found TR root extract to have antiproliferative activity on glioma cells after 24 h of treatment. TR root extract induces apoptosis on various grades (I-III) of human glioma cells by the generation of reactive oxygen species (ROS) along with concurrent loss of mitochondrial membrane potential, enhanced S and G2/M phases of the cell cycle, and altered mRNA levels of Bax, Bcl-2, p53, Cas-3, Cas-8 and Cas-9 factors involved in apoptosis. This work for the first time demonstrate that TR extract from L. sibiricus root has the potential to activate apoptosis in grade I-III human glioma cells through the intrinsic and extrinsic pathways.

Phenolic constituents from Wissadula periplocifolia (L.) C. Presl. and anti-inflammatory activity of 7,4'-di-O-methylisoscutellarein.

This study reports the first phenolics from Wissadula genus (Malvaceae) and the anti-inflammatory activity of 7,4'-di-O-methylisoscutellarein. Using chromatographic methods, five phenolic compounds were isolated from aerial parts of Wissadula periplocifolia (L.) C. Presl. The compounds were identified as 4-hydroxybenzoic acid, 3-hydroxybenzoic acid, trans-cinnamic acid, tamgermanetin and 7,4'-di-O-methylisoscutellarein using spectroscopic methods. The flavone 7,4'-di-O-methylisoscutellarein showed anti-inflammatory activity by inhibiting neutrophils recruitment in a mice model of pleurisy and by decreasing significantly the production of cytokines IL-1ß and TNF-α.

Protective effect of Tremella fuciformis Berk extract on LPS-induced acute inflammation via inhibition of the NF-κB and MAPK pathways.

Tremella fuciformis Berk (TFB) has long been used as a traditional medicine in Asia. Although TFB exhibits antioxidant and anti-inflammatory effects, the mechanisms of action responsible have remained unknown. We confirmed the anti-inflammatory effects of Tremella fuciformis Berk extract (TFE) in RAW 264.7 cells and observed significantly suppressed LPS-induced iNOS/NO and COX-2/PGE2 production. TFE also suppressed LPS-induced IKK, IkB, and p65 phosphorylation, as well as LPS-induced translocation of p65 from the cytosol. Additionally, TFE inhibited LPS-induced phosphorylation of MAPKs. In an acute inflammation study, oral administration of TFE significantly inhibited LPS-induced IL-1ß, IL-6 and TNF-α production and iNOS and COX-2 expression. The major bioactive compounds from TFB extract were identified as gentisic acid, protocatechuic acid, 4-hydroxybenzoic acid, and coumaric acid. Among these compounds, protocatechuic acid showed the strongest inhibitory effects on LPS-induced NO production in RAW 264.7 cells. Overall, these results suggest that TFE is a promising anti-inflammatory agent that suppresses iNOS/NO and COX-2/PGE2 expression, as well as the NF-κB and MAPK signaling pathways.

Antibacterial Constituents of Hainan Morinda citrifolia (Noni) Leaves.

Noni (Morinda citrifolia L.) is an edible and medicinal plant distributed in Hainan, China. The antibacterial activities of the extracts of water (WE), petroleum ether (PEE), ethyl acetate (EAE), chloroform (CE), and n-butanol (BE) were assayed by the disk diffusion method. The results showed that the extracts from Noni leaves possessed antibacterial effects against Bacillus subtilis, Escherichia coli, Proteus vulgaris, and Staphylococcus aureus. Among 5 different extracts, the BE produced the best antibacterial activity. The samples were first extracted by ethanol, and the primary compounds in the BE fraction of ethanol extract was further isolated and identified. Six phenolic compounds, including 5, 15-dimethylmorindol, ferulic acid, p-hydroxycinamic acid, methyl 4-hydroxybenzoate, methyl ferulate, and methyl 4-hydroxycinnamate, were identifiedby NMR. The results indicated that the phenolic compounds might significantly contribute to antibacterial activities of Noni leaves.

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In order to investigate the relationship between the accumulation of phenolic acids in peanut continuous cropping soil and the continuous cropping obstacle of peanut, the effects of p-hydroxy benzoic acid and cinnamic acid on peanut root zone soil nutrients, soil enzyme activities and yield of peanut were studied by pot experiment at three stages of peanut, i.e. the pegging stage of peanut (45 days after seedling), the early podding (75 days after seedling) and the end of podding (105 days after seedling) stages. The results showed that the peanut root zone soil nutrients and enzyme activities changed obviously under the two phenolic acids treatment, especially at the pegging stage of peanut. The soil alkali-hydrolyzable nitrogen, available phosphorus, available potassium, and soil enzyme activities (urease, sucrose, neutral phosphatase) were decreased significantly. At the early and end of podding stages of peanut, the effects of the two phenolic acids on peanut root zone soil nutrients and soil enzyme activities were under a weakening trend. The allelopathy of cinnamic acid was stronger than that of p-hydroxy benzoic acid at the same initial content. The pod yield per pot was reduced by 45.9% and 52.8%, while the pod number of per plant was reduced by 46.2% and 48.9% at higher concentration (80 mg·kg-1 dry soil) of p-hydroxy benzoic acid and cinnamic acid treatments, respectively.

Asplenioideae Species as a Reservoir of Volatile Organic Compounds with Potential Therapeutic Properties.

Twelve French Asplenioideae ferns (genera Asplenium and subgenera Ceterach and Phyllitis) were investigated for the first time for volatile organic compounds (VOC) using GC-MS. Sixty-two VOC biosynthesized from the lipidic, shikimic, terpenic and carotenoid pathways were identified. Several VOC profiles can be highlighted from Asplenium jahandiezii and A. xalternifolium with exclusively lipidic derivatives to A. onopteris with an equal ratio of lipidic/shikimic compounds. Very few terpenes as caryophyllene derivatives were identified, but only in A. obovatum subsp. bilotii. The main odorous lipidic derivatives were (E)-2-decenal (waxy and fatty odor), nonanal (aldehydic and waxy odor with a fresh green nuance), (E)-2-heptenal (green odor with a fatty note) and 1-octen-3-ol (mushroom-like odor), reported for all species. A few VOC are present in several species in high content, i.e., 9-oxononanoic acid used as a precursor for biopolymers (19% in A. jahandiezii), 4-hydroxyacetophenone with a sweet and heavy floral odor (17.1% in A. onopteris), and 4-hydroxybenzoic acid used as a precursor in the synthesis of parabens (11.3% in A. foreziense). Most of the identified compounds have pharmacological activities, i.e., octanoic acid as antimicrobial, in particular against Salmonellas, with fatty and waxy odor (41.1% in A. petrarchae), tetradecanoic acid with trypanocidal activity (13.3% in A. obovatum subsp. bilotii), 4-hydroxybenzoic acid (8.7% in A. onopteris) with antimicrobial and anti-aging effects, 3,4-dihydroxybenzaldehyde as an inhibitor of growth of human cancer cells (6.7% in Ceterach officinarum), and phenylacetic acid with antifungal and antibacterial activities (5.8% in A. onopteris). Propionylfilicinic acid was identified in the twelve species. The broad spectrum of odorous and bioactive VOC identified from the Asplenium, Ceterach and Phyllitis species are indeed of great interest to the cosmetic and food industries.

8-p-Hdroxybenzoyl Tovarol Induces Paraptosis Like Cell Death and Protective Autophagy in Human Cervical Cancer HeLa Cells.

8-p-Hdroxybenzoyl tovarol (TAW) is a germacrane-type sesquiterpenoid that can be isolated from the roots of Ferula dissecta (Ledeb.) Ledeb. In this study, the growth inhibitory effects induced by TAW were screened on some types of tumor cells, and the mechanism was investigated on TAW-induced growth inhibition, including paraptosis and autophagy in human cervical cancer HeLa cells. TAW-induced paraptosis involved extensive cytoplasmic vacuolization in the absence of caspase activation. Additionally, TAW evoked cell paraptotic death mediated by endoplasmic reticulum (ER) stress and unfolded protein response (UPR). Autophagy induced by TAW was found to antagonize paraptosis in HeLa cells. This effect was enhanced by rapamycin and suppressed by the autophagy inhibitor, 3-methyladenine (3MA). Loss of beclin 1 (an autophagic regulator) function led to promote ER stress. Taken together, these results suggest that TAW induces paraptosis like cell death and protective autophagy in HeLa cells, which would provide a new clue for exploiting TAW as a promising agent for the treatment of cervical cancer.

Characterization of in vitro antifungal activities of small and American cranberry (Vaccinium oxycoccos L. and V. macrocarpon Aiton) and lingonberry (Vaccinium vitis-idaea L.) concentrates in sugar reduced fruit spreads.

In this study, cranberry and lingonberry concentrates were added to commercial sugar-reduced fruit spreads (raspberry-Aloe vera, strawberry-guava, and strawberry-lime), and tested for their antifungal activities. Selected strains of the species Absidia glauca, Penicillium brevicompactum, Saccharomyces cerevisiae and Zygosaccharomyces bailii, as well as xerophilic environmental isolates of the genera Penicillium and Eurotium were used for challenge testing. Initially, varying concentrations of synthetic antifungal agents, such as sodium benzoate, potassium sorbate and butyl 4-hydroxybenzoate were tested against these fungi on wort agar containing 31% fructose at different pH values. Subsequently, the experiments were conducted in fruit spreads containing different concentrations of cranberry and lingonberry concentrates. The results of this study demonstrate that these concentrates were able to inhibit growth of visible colonies of xerophilic and non-xerophilic fungi. Cranberry and lingonberry concentrates are interesting candidates for natural preservation against fungal growth in sugar reduced fruit spreads.

Screening of bisphenol A, triclosan and paraben analogues as modulators of the glucocorticoid and androgen receptor activities.

A homeostasis of the glucocorticoid and androgen endocrine system is essential to human health. Their disturbance can lead to various diseases, for example cardiovascular, inflammatory and autoimmune diseases, infertility, cancer. Fifteen widely used industrial chemicals that disrupt endocrine activity were selected for evaluation of potential (anti)glucocorticoid and (anti)androgenic activities. The human breast carcinoma MDA-kb2 cell line was utilized for reporter gene assays, since it expresses both the androgen and the glucocorticoid-responsive reporter. Two new antiandrogens, 4,4'-sulfonylbis(2-methylphenol) (dBPS) and 4,4'-thiodiphenol (THIO), and two new antiglucocorticoids, bisphenol Z and its analog bis[4-(2-hydroxyethoxy)phenyl] sulfone (BHEPS) were identified. Moreover, four new glucocorticoid agonists (methyl paraben, ethyl paraben, propyl paraben and bisphenol F) were found. To elucidate the structure-activity relationship of bisphenols, we performed molecular docking experiments with androgen and glucocorticoid receptor. These docking experiments had shown that bulky structures such as BHEPS and bisphenol Z act as antiglucocorticoid, because they are positioned toward helix H12 in the antagonist conformation and could therefore be responsible for H12 conformational change and the switch between agonistic and antagonistic conformation of receptor. On the other hand smaller structures cannot interact with H12. The results of in vitro screening of fifteen industrial chemicals as modulators of the glucocorticoid and androgen receptor activities demand additional in vivo testing of these chemicals for formulating any relevant hazard identification to human health.

Synthesis and characterization of silver nanoparticles using crystal compound of sodium para-hydroxybenzoate tetrahydrate isolated from Vitex negundo. L leaves and its apoptotic effect on human colon cancer cell lines.

Metallic nanoparticles are major concern, particularly silver nanoparticles (AgNPs) are used in various applications. In the present investigation, we report a novel strategy with biological approach for synthesis of AgNPs using sodium para-hydroxybenzoate tetrahydrate (SPHT) isolated from Vitex negundo leaves. The synthesized SPHT-AgNPs were characterized by UV-vis spectroscopy, high resolution transmission electron microscopy (HRTEM) with selected area electron diffraction (SAED) pattern, field emission scanning electron microscopy (FESEM) with energy-dispersive X-ray spectroscopy (EDX), zeta potential and Fourier transform infrared spectroscopy (FT-IR) analysis. The various pH and temperature were evaluated to find their stability effects on SPHT-AgNPs synthesis peak at 430 nm. The size of SPHT-AgNPs were ranging from 26 to 39 nm and were spherical in shape. The hydroxyl and carboxylic functional groups from bio-reducing mediators of SPHT have a stronger ability towards synthesis of AgNPs, which was confirmed using FT-IR spectrum. In addition, anticancer activity were determined by MTT assay, Annexin V-FITC/PI and cell cycle analysis.

Hydrolytic enzymes production by Aspergillus section Nigri in presence of butylated hydroxyanisole and propyl paraben on peanut meal extract agar.

BACKGROUND: In the last years, food grade antioxidants are used safely as an alternative to traditional fungicides to control fungal growth in several food and agricultural products. AIMS: In this work, the effect of butylated hydroxyanisole (BHA) and propyl paraben (PP) on two hydrolytic enzyme activity (ß-d-glucosidase and α-d-galactosidase) by Aspergillus section Nigri species under different water activity conditions (aW; 0.98, 0.95 and 0.93) and incubation time intervals (24, 48, 72 and 96h) was evaluated on peanut-based medium. METHODS: The activity of two glycosidases, ß-d-glucosidase and α-d-galactosidase, was assayed using as substrates 4-nitrophenyl-ß-d-glucopyranosido and 4-nitrophenyl-α-d-galactopyranosido, respectively. The enzyme activity was determined by the increase in optical density at 405nm caused by the liberation of p-nitrophenol by enzymatic hydrolysis of the substrate. Enzyme activity was expressed as micromoles of p-nitrophenol released per minute. RESULTS: The major inhibition in ß-d-glucosidase activity of A. carbonarius and A. niger was found with 20mmoll(-1) of BHA or PP at 0.98 and 0.95 aW, respectively, whereas for α-d-galactosidase activity a significant decrease in enzyme activity with respect to control was observed in A. carbonarius among 5 to 20mmoll(-1) of BHA or PP in all conditions assayed. Regarding A. niger, the highest percentages of enzyme inhibition activity were found with 20mmoll(-1) of BHA or PP at 0.95 aW and 96h. CONCLUSIONS: The results of this work provide information about the capacity of BHA and PP to inhibit in vitro conditions two of the most important hydrolytic enzymes produced by A. carbonarius and A. niger species.

Paraben resistance in bacteria from sewage treatment plant effluents in India.

Parabens, the antimicrobial preservatives used in cosmetics, food and pharmaceuticals, are often detected in the aquatic environment. Generally, sewage treatment plants (STPs) receive community sewage containing parabens, which are ultimately released into streams/rivers. In this study, bacteria in STP effluents were evaluated for their resistance to parabens. The susceptibility was in the order of Staphylococcus aureus > Bacillus sp. >Escherichia coli > Pseudomonas aeruginosa. Gram-negative bacteria showed less susceptibility than their control and Gram-positive bacteria. Further, the bacteria were more sensitive towards butyl and ethyl parabens. Interestingly, the strains showed resistance to ≥5 mg of parabens, which is equivalent to or higher than reported environmental concentrations. The increase in paraben chain length did not enhance the susceptibility in all cases and it was understood that the activity may differ for each bacterium in the environment. This is the first profile on paraben resistance in common pathogens of Indian STPs. Paraben resistance may be developed due to continuous exposure even at sub-inhibitory and/or chronic levels in the environment and this resistance may be transferred to other pathogenic bacteria in receiving waters. Thus the study demonstrates the effectiveness of the disc diffusion method in environmental bacterial resistance assessment and addresses the risk involved in the use of parabens.

Solid lipid nanoparticles as effective reservoir systems for long-term preservation of multidose formulations.

Cosmetic multidose preparations, as well as pharmaceutical ones, are at risk of contamination by microorganisms, due to their high water content. Besides the risk of contamination during manufacturing, multidose cosmetic preparations may be contaminated by consumers during their use. In this paper, the results of the utilization of nanoparticles as reservoir systems of parabens, the most used class of preservatives, were reported. Two different systems, solid lipid nanoparticles (SLN) made of pure precirol and nanostructured lipid carriers (NLC) made of precirol and almond oil, containing three parabens as single molecules or as a mixture, were prepared and tested. All the systems were characterized for size, polydispersion index, zeta potential and encapsulation efficiency. Release experiments, carried out in steady state and sink conditions, allowed to evidence that both SLN and NLC were able to act as reservoir systems. The antimicrobial activity of the systems was tested against Candida albicans ATCC 10231 with repeat insult tests. The results of the release experiments and the antimicrobial tests showed very low water concentration of parabens still maintaining their antimicrobial activity.

UPLC-PDA-ESI-MS/MS analysis of compounds extracted by cardiac h9c2 cell from Polygonum orientale.

INTRODUCTION: A flavonoid-enriched extract (FEE) of Polygonum orientale was reported to show cardioprotective effect but only very few compounds were reported to contribute to the effect. Identification of compounds interacting with the target cardiac cell is important for the understanding of active compounds. OBJECTIVE: To develop an efficient method for the screening of potential active compounds directly acting on the target cardiac cell in FEE and to structurally characterise these compounds. METHODOLOGY: Flavonoid-enriched extract was prepared by extraction of the plant with water, addition of ethanol to the solution to remove polysaccharides and proteins, and removal of tannins by a polyamide column chromatography. Cell extraction was conducted on a cardiac h9c2 cell and the solution containing compounds released from the cell were desalted by solid phase extraction. Compounds present in the cell extract were detected by ultra-performance liquid chromatography (UPLC) and targeted multi-reaction monitoring (MRM), while their structures were characterised by UPLC-photodiodide array (PDA)-electrospray ion source (ESI)-MS/MS investigations of the FEE. RESULTS: Twenty-three potentially active phenolics including ten flavonoid C-glycosides and six flavonoid O-glycosides have been identified from the 40 compounds screened in the cell extract. Among these compounds, three were new and nine were identified from this plant for the first time. Strategies for the structural characterisation of flavonoid glycosides were also discussed. CONCLUSION: The study has shown that FEE contains the flavonoid as its major principles and the coupling of UPLC-PDA-ESI-MS/MS and targeted UPLC-MRM with target cell extraction is an efficient method for the screening and structural characterisation of potential active compounds.

Nematotoxic phenolic compounds from Melia azedarach against Meloidogyne incognita.

In the present study, evaluated was the paralysis activity of whole Italian and Algerian Melia azedarach, commonly known as chinaberry, fruits and parts (seeds, wood, and kernels) against Meloidogyne incognita second stage juveniles (J(2)). The paralysis activity was evaluated in vitro after 1 h and 1 day immersion periods of nematodes in test solutions. Phenolic constituent components of the extracts were identified and quantified by high-performance liquid chromatography-mass spectrometry, while confirmation was performed by high-performance liquid chromatography-diode array. The water extract of the Italian M. azedarach fruit pulp (IPWE) showed significant nematicidal activity (EC(50/48h) = 955 µg/mL) and among its active ingredient components were p-coumaric acid and p-hydroxybenzoic acid (EC(50/48h) = 840 and 871 µg/mL, respectively). This is the first report of the nematicidal activity of M. azedarach pulp water extract and phenolic acids against the root knot nematode M. incognita.