Comprehensive Analysis of NAC Transcription Factors Reveals Their Evolution in Malvales and Functional Characterization of AsNAC019 and AsNAC098 in Aquilaria sinensis.

NAC is a class of plant-specific transcription factors that are widely involved in the growth, development and (a)biotic stress response of plants. However, their molecular evolution has not been extensively studied in Malvales, especially in Aquilaria sinensis, a commercial and horticultural crop that produces an aromatic resin named agarwood. In this study, 1502 members of the NAC gene family were identified from the genomes of nine species from Malvales and three model plants. The macroevolutionary analysis revealed that whole genome duplication (WGD) and dispersed duplication (DSD) have shaped the current architectural structure of NAC gene families in Malvales plants. Then, 111 NAC genes were systemically characterized in A. sinensis. The phylogenetic analysis suggests that NAC genes in A. sinensis can be classified into 16 known clusters and four new subfamilies, with each subfamily presenting similar gene structures and conserved motifs. RNA-seq analysis showed that AsNACs presents a broad transcriptional response to the agarwood inducer. The expression patterns of 15 AsNACs in A. sinensis after injury treatment indicated that AsNAC019 and AsNAC098 were positively correlated with the expression patterns of four polyketide synthase (PKS) genes. Additionally, AsNAC019 and AsNAC098 were also found to bind with the AsPKS07 promoter and activate its transcription. This comprehensive analysis provides valuable insights into the molecular evolution of the NAC gene family in Malvales plants and highlights the potential mechanisms of AsNACs for regulating secondary metabolite biosynthesis in A. sinensis, especially for the biosynthesis of 2-(2-phenyl) chromones in agarwood.

Selectivity Tuning by Natural Deep Eutectic Solvents (NADESs) for Extraction of Bioactive Compounds from Cytinus hypocistis-Studies of Antioxidative, Enzyme-Inhibitive Properties and LC-MS Profiles.

In the present study, the extracts of Cytinus hypocistis (L.) L using both traditional solvents (hexane, ethyl acetate, dichloromethane, ethanol, ethanol/water, and water) and natural deep eutectic solvents (NADESs) were investigated in terms of their total polyphenolic contents and antioxidant and enzyme-inhibitive properties. The extracts were found to possess total phenolic and total flavonoid contents in the ranges of 26.47-186.13 mg GAE/g and 0.68-12.55 mg RE/g, respectively. Higher total phenolic contents were obtained for NADES extracts. Compositional differences were reported in relation to antioxidant potential studied by several assays (DPPH: 70.19-939.35 mg TE/g, ABTS: 172.56-4026.50 mg TE/g; CUPRAC: 97.41-1730.38 mg TE/g, FRAP: 84.11-1534.85 mg TE/g). Application of NADESs (choline chloride-urea 1:2, a so-called Reline) allowed one to obtain the highest number of extracts having antioxidant potential in the radical scavenging and reducing assays. NADES-B (protonated by HCl L-proline-xylitol 5:1) was the only extractant from the studied solvents that isolated a specific fraction without chelating activity. Reline extract exhibited the highest acetylcholinesterase inhibition compared to NADES-B and NADES-C (protonated by H2SO4 L-proline-xylitol 5:1) extracts, which showed no inhibition. The NADES extracts were observed to have higher tyrosinase inhibitory properties compared to extracts obtained by traditional organic solvents. Furthermore, the NADES extracts were relatively better inhibitors of the diabetic enzymes. These findings provided an interesting comparison in terms of total polyphenolic content yields, antioxidant and enzyme inhibitory properties (cholinesterase, amylase, glucosidase, and tyrosinase) between traditional solvent extracts and NADES extracts, used as an alternative. While the organic solvents showed better antioxidant activity, the NADES extracts were found to have some other improved properties, such as higher total phenolic content and enzyme-inhibiting properties, suggesting functional prospects for their use in phytonutrient extraction and fractionation. The obtained results could also be used to give a broad overview of the different biological potentials of C. hypocistis.

In Vitro Anti-Diabetic Activities and UHPLC-ESI-MS/MS Profile of Muntingia calabura Leaves Extract.

Anti-diabetic compounds from natural sources are now being preferred to prevent or treat diabetes due to adverse effects of synthetic drugs. The decoction of Muntingia calabura leaves was traditionally consumed for diabetes treatment. However, there has not been any published data currently available on the processing effects on this plant's biological activity and phytochemical profile. Therefore, this study aims to evaluate the effect of three drying methods (freeze-drying (FD), air-drying (AD), and oven-drying (OD)) and ethanol:water ratios (0, 50, and 100%) on in vitro anti-diabetic activities of M. calabura leaves. In addition, an ultrahigh-performance-liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) method was used to characterize the metabolites in the active extract. The FD M. calabura leaves, extracted with 50% ethanol, is the most active extract that exhibits a high α-glucosidase and α-amylase inhibitory activities with IC50 values of 0.46 ± 0.05 and 26.39 ± 3.93 µg/mL, respectively. Sixty-one compounds were tentatively identified by using UHPLC-ESI-MS/MS from the most active extract. Quantitative analysis, by using UHPLC, revealed that geniposide, daidzein, quercitrin, 6-hydroxyflavanone, kaempferol, and formononetin were predominant compounds identified from the active extract. The results have laid down preliminary steps toward developing M. calabura leaves extract as a potential source of bioactive compounds for diabetic treatment.

Fractionated Extracts From Gnidia kraussiana (Malvales: Thymeleaceae) as Bioactive Phytochemicals for Effective Management of Callosobruchus maculatus (Coleoptera: Chrysomelidae) in Stored Vigna unguiculata (Fabales: Fabaceae) Seeds.

One of the most important global problems is protecting food from insect pests. The negative effects of synthetic insecticides on human health led to a resurgence of interest in botanical insecticides due to their minimal ecological side effects. Therefore, the insecticidal potential of hexane, acetone, and methanol extracts of Gnidia kraussiana Meisn roots at 1 and 5g/kg, and neem seed oil (NSO), used as standard insecticide, were evaluated. Ovicidal and larvicidal toxicity was tested by treating freshly laid eggs and larvae at different immature stages of Callosobruchus maculatus (F.). Cowpea (Vigna unguiculata) (L.) Walp seed damage and weight loss were assessed after a storage period of 4 mo. Repellency effects were detected in choice test using a linear olfactometer. All the fractions were toxic to C. maculatus; however, their bioactivities were inversely correlated with products polarity. Extracts proved to be more toxic than the commercial NSO. The acetone extract was more effective against immature stages of C. maculatus than the methanol extract; eggs, first-, and second-instar larvae being the more susceptible. No cowpea seed damage and weight loss were recorded from the seeds treated with hexane and acetone extracts at the dosage of 5 g/kg, after 4 mo of storage. Extracts evoked stronger repellency effects compared with the tested standard insecticide. According to the above, hexane and acetone extracts are good candidates for incorporation in integrated pest management programs for the control of C. maculatus in stored cowpea seeds.

Fermentation of Jamaican Cherries Juice Using Lactobacillus plantarum Elevates Antioxidant Potential and Inhibitory Activity against Type II Diabetes-Related Enzymes.

Jamaican cherry (Muntinga calabura Linn.) is tropical tree that is known to produce edible fruit with high nutritional and antioxidant properties. However, its use as functional food is still limited. Previous studies suggest that fermentation with probiotic bacteria could enhance the functional properties of non-dairy products, such as juices. In this study, we analyze the metabolite composition and activity of Jamaican cherry juice following fermentation with Lactobacillus plantarum FNCC 0027 in various substrate compositions. The metabolite profile after fermentation was analyzed using UPLC-HRMS-MS and several bioactive compounds were detected in the substrate following fermentation, including gallic acid, dihydrokaempferol, and 5,7-dihydroxyflavone. We also found that total phenolic content, antioxidant activities, and inhibition of diabetic-related enzymes were enhanced after fermentation using L. plantarum. The significance of its elevation depends on the substrate composition. Overall, our findings suggest that fermentation with L. plantarum FNCC 0027 can improve the functional activities of Jamaican cherry juice.

Study on Gas Chromatographic Fingerprint of Essential Oil from Stellera chamaejasme Flowers and Its Repellent Activities against Three Stored Product Insects.

The objective of this study was to establish the chromatographic fingerprints of the essential oil (EO) from Stellera chamaejasme flowers collected from various natural sites by gas chromatography (GC) combined with chemometric methods. The EO was obtained by hydrodistillation, and its chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Most components were identified as ketones and the relatively high-content components were fitone (38.973%), n-hentriacontane (5.807%), myristic acid (4.944%) and phytol (3.988%). In addition, the repellent activities of the EO from S. chamaejasme flowers and its four main chemical compounds were evaluated against three stored product pests (Tribolium castaneum, Lasioderma serricorne, Liposcelis bostrychophila) for the first time. In this work, the EO and the four chemical compounds showed a repellent effect against three storage pests after 2 and 4 h exposure. The experimental method and repellent activity of S. chamaejasme flower EO could provide a basis for the development of botanical pesticide and the utilization of the rich plant resources of S. chamaejasme in the future.

Phenolic Constituents of the Roots of Rhamnoneuron balansae with Senolytic Activity.

With the advent of senolytic agents capable of selectively removing senescent cells in old tissues, the perception of age-associated diseases has been changing from being an inevitable to a preventable phenomenon of human life. In the search for materials with senolytic activity from natural products, six new flavonostilbenes (1-6), three new phenylethylchromanones (7-9), three new phenylethylchromones (10-12), and four known compounds (13-16) were isolated from the roots of Rhamnoneuron balansae. The chemical structures of these isolated compounds were determined based on the interpretation of spectroscopic data, including 1D and 2D NMR, ECD, and HRMS. The absolute configuration of compound 1 was also determined by a Mosher ester analysis and ECD calculations. Compounds 6-8 were shown to selectively destroy senescent cells, and the promoter activity of p16INK4A, a representative senescence marker, was reduced significantly by compound 6. The present results suggest the potential activity of flavonostilbene and phenylethylchromanone skeletons from R. balansae as new senolytics.

Safety evaluation of orally-administered methanol extract of Muntingia calabura Linn. leaves: A sub-chronic toxicity study in Sprague Dawley rats.

Muntingia calabura (M. calabura), locally known as "kerukup siam" or "buah ceri" belongs to the family Muntingiaceae and has been scientifically demonstrated to exert various pharmacological activities. The objectives of the current study are to evaluate the antioxidant activities and to determine the subchronic toxicity of 90 days orally-administered methanol extract of M. calabura (MEMC) in male Sprague Dawley rats. The rats were randomly divided into four groups (n=6). Vehicle control received 8% tween 80 and treatment group received 50, 250 and 500 mg/kg of MEMC orally administered daily for 90 days. Blood collection was carried out to obtain the hematological and biochemical profile of the rats. The organs harvested were subjected to histopathological analysis. For the antioxidant test, the extract was subjected to antioxidant study using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH)- and superoxide anion-radical scavenging activity, total phenolic content (TPC) and phytochemical screening. Results obtained show that no adverse effects were observed during the experimental period. Hematological and biochemical analysis also showed no significant changes in this toxicity study. Besides, antioxidant analyses revealed that MEMC has higher DPPH- and SOD-radical scavenges activity as well as higher TPC value. In conclusion, M. calabura is safe for consumption and possesses beneficial antioxidant effect.

Stellera chamaejasme and Its Main Compound Luteolin 7-O-Glucoside Alleviates Skin Lesions in Oxazolone- and 2,4-Dinitrochlorobenzene-Stimulated Murine Models of Atopic Dermatitis.

Stellera chamaejasme, also known as "Langdu", has been traditionally used for the management of skin-related diseases such as psoriasis and skin ulcers. The aim of this study was to determine whether S. chamaejasme and its major component, luteolin 7-O-glucoside, have a preventive effect on the development of atopic dermatitis in oxazolone-treated BALB/c mice and 2,4-dinitrochlorobenzene-treated hairless mice. The epicutaneous applications of oxazolone and 2,4-dinitrochlorobenzene evoke an experimental murine atopic dermatitis-like reaction in BALB/c mouse ears and SKH-1 hairless mice. Atopic skin symptoms, including erythema (redness), pruritus (itching), exudation (weeping), excoriation (peeling), and lichenification (skin thickening), responded to treatment with S. chamaejasme aerial parts EtOH extract for 2 or 3 weeks. Histopathological examination revealed S. chamaejasme aerial parts EtOH extract significantly reduced inflammatory cell infiltration when applied to atopic dermatitis mice. In addition, luteolin 7-O-glucoside, the major active compound of the S. chamaejasme aerial parts EtOH extract, decreased serum IgE and IL-4 levels and transepidermal water loss and increased skin hydration, therefore exhibiting strong anti-atopic dermatitis activity in 2,4-dinitrochlorobenzene-induced atopic dermatitis mice. In this study, we confirmed antipruritic and antidermatitic effects of S. chamaejasme extract and its main component luteolin 7-O-glucoside in atopic dermatitis murine models. The study shows S. chamaejasme aerial parts EtOH extract and luteolin 7-O-glucoside are most likely to be potential drug candidates for atopic dermatitis treatment.

Tannin profile, antioxidant properties, and antimicrobial activity of extracts from two Mediterranean species of parasitic plant Cytinus.

BACKGROUND: Cytinus is small genus of endophytic parasitic plants distributed in South Africa, Madagascar, and in the Mediterranean region. In the latter area, two species occur, Cytinus hypocistis and C. ruber, distinguished by both morphological characters and ecological traits. We characterized the ethanolic and aqueous extracts obtained from the inflorescences of C. hypocistis and C. ruber collected in Sardinia, Italy, and explored their tannin content, antioxidant properties and antimicrobial activities. METHODS: Total phenolic contents were determined by Folin-Ciocalteu spectrophotometric method. Tannin content was determined by HPLC. Antioxidant activity of the extracts was tested with both electron transfer-based (FRAP, TEAC, DPPH) and spectrophotometric HAT methods (ORAC-PYR). The antimicrobial activities of extracts/compounds were evaluated using the broth microdilution method. The bactericidal activity was evaluated using the time-kill method. Biofilm formation was evaluated by crystal violet (CV) staining assay. RESULTS: Characterization of the tannin profile of C. hypocistis and C. ruber revealed a significant amount of gallotannins, in particular 1-O-galloyl-ß-D-glucose. In addition, pentagalloyl-O-ß-D-glucose was present in all extracts, reaching the concentration of 0.117 g/kg in the ethanolic extract of C. hypocistis. C. hypocistis extracts displayed a strongest antioxidant activity than C. ruber extracts. Three Gram-positive bacterial species tested (Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecium) resulted sensitive to both Cytinus extracts, with MICs ranging from 125 to 500 µg/ml for aqueous extracts and from 31.25 to 250 µg/ml for ethanolic extracts; on the contrary, Gram-negative strains (Pseudomonas aeruginosa and Klebsiella pneumoniae) were not affected by Cytinus extracts. Intriguingly, we observed the suppressive activity of ethanolic extracts of C. hypocistis and C. ruber on biofilm formation of S. epidermidis. Experiments performed with synthetic compounds indicated that pentagalloyl-O-ß-D-glucose is likely to be one of the active antimicrobial components of Cytinus extracts. CONCLUSIONS: These findings show that Cytinus extracts have antimicrobial and antioxidant activities, suggesting a possible application of Cytinus as sources of natural antimicrobials and antioxidants.

Anticancer Potential of Fruit Extracts from Vatica diospyroides Symington Type SS and Their Effect on Program Cell Death of Cervical Cancer Cell Lines.

Vatica diospyroides Symington is locally known as Chan-Ka-Pho in Thailand. Ancient people have used it as therapeutic plant for cardiac and blood tonic cure. The purpose of this study was to investigate the potential cytotoxicity and selectivity of the extracts from V. diospyroides type SS fruit on cervical cancer HeLa and SiHa cell lines and to examine its underlying mechanism of action. MTT assay revealed that the extracts showed inhibition of cell survival in a dose-dependent manner and exhibited highly cytotoxic activity against both HeLa and SiHa cells with IC50 value less than 20 µg/mL along with less toxicity against L929 cells. Acetone cotyledon extract (ACE) showed the best selectivity index value of 4.47 (HeLa) and 3.51 (SiHa). Distinctive morphological changes were observed in ACE-treated cervical cancer cells contributing to apoptosis action. Flow cytometry analysis with Annexin V-FITC and PI staining precisely indicated that ACE induced apoptosis in HeLa and SiHa cell lines in a dose-dependent manner. Treatment of ACE with half IC50 caused DNA fragmentation and also activated increasing of bax and cleaved caspase-8 protein in HeLa cells after 48 h exposure. The results suggest that ACE has potent and selective cytotoxic effect against cervical cancer cells and the potential to induce bax and caspase-8-dependent apoptosis. Hence, the ACE could be further exploited as a potential lead in cancer treatment.

Methanol extract of Muntingia calabura leaves attenuates CCl4-induced liver injury: possible synergistic action of flavonoids and volatile bioactive compounds on endogenous defence system.

CONTEXT: Muntingia calabura L. (Muntingiaceae) exerts antioxidant and anti-inflammatory activities, thus, it might be a good hepatoprotective agent. OBJECTIVE: This study investigates the effect of methanol extract of M. calabura leaves (MMCL) on hepatic antioxidant and anti-inflammatory activities in CCl4-induced hepatotoxic rat. MATERIALS AND METHODS: Sprague Dawley rats (n = 6) were treated (p.o.) with 10% DMSO (Groups 1 and 2), 50 mg/kg N-acetylcysteine (Group 3) or, 50, 250, or 500 mg/kg MMCL (Groups 4-6) for 7 consecutive days followed by pretreatment (i.p.) with vehicle (Group 1) or 50% CCl4 in olive oil (v/v) (Groups 2-6) on day 7th. Plasma liver enzymes and hepatic antioxidant enzymes and pro-inflammatory cytokines concentrations were measured while liver histopathology was examined. RESULTS: MMCL, at 500 mg/kg, significantly (p < 0.05) ameliorated CCl4-induced hepatotoxicity by decreasing the plasma level of alanine transaminase (429.1 versus 168.7 U/L) and aspartate transaminase (513.8 versus 438.1 U/L) as well as the tissue level of nitric oxide (62.7 versus 24.1 nmol/g tissue). At 50, 250, or 500 mg/kg, MMCL significantly (p < 0.05) reduced the tumour necrosis factor α (87.8 versus 32.7 pg/mg tissue), interleukin-1ß (1474.4 versus 618.3 pg/mg tissue), and interleukin-6 (136.7 versus 30.8 pg/mg tissue) while increased the liver catalase (92.1 versus 114.4 U/g tissue) and superoxide dismutase (3.4 versus 5.5 U/g tissue). Additionally, qualitative phytochemicals analysis showed that MMCL contained gallic acid, ferulic acid, quercetin, and genistein. DISCUSSION AND CONCLUSIONS: MMCL ability to attenuate CCl4-induced hepatotoxicity could be helpful in the development of hepatoprotective agents with fewer side effects.

Pterospermum acerifolium (L.) wild bark extract induces anticarcinogenic effect in human cancer cells through mitochondrial-mediated ROS generation.

Plants have many medicinal properties including anticancer activity due to the presence of several secondary metabolites. Current cancer treatment policies are not much effective because of side effects and resistance development. Therefore, the discovery of new phytotherapeutics with no or fewer side effects is highly needed. Pterospermum acerifolium (L.) wild, an angiosperm has a broad application in traditional Indian medicinal system including cancer treatment. Despite, there is no study available on the cytotoxic and apoptotic effect of P. acerifolium in human cancer cells. Exploring the medicinal properties of P. acerifolium plant by its traditional use will be helpful towards developing novel cancer therapeutics. Hence, we decided to demonstrate the anti-carcinogenic property of P. acerifolium ethanolic bark extract against lung (A549) and pancreatic (PANC-1) cancer cells. The cytotoxicity was demonstrated by MTT assay, morphological changes, and scratch invasion assay. Flow cytometry, fluorescence staining techniques, and cell cycle analysis were confirmed the apoptotic property of P. acerifolium plant. The cell viability assay revealed that P. acerifolium ethanolic bark extract significantly reduced the viability of both A549 and PANC-1 cells. Moreover, PANC-1 cells showed more sensitivity towards P. acerifolium ethanolic bark extract than A549 at higher concentrations. Clear visualization of changes such as cytoplasmic condensation, cellular morphology, cell shrinkage, and augmented number of dead cells in both the cancer cells was observed after treatment. Scratch and invasion assay showed that cell migration and invasion rate of both the cancer cells were significantly reduced. Fluorescence microscopic studies using acridine orange/ethidium bromide and DAPI (4', 6-diamidino-2-phenylindole) staining showed early and late apoptotic symptoms after treatment with bark extract. Rhodamine-123 and DCFH-DA staining analysis by fluorescence and flow cytometry showed that bark extract depolarized the mitochondria membrane potential and induced reactive oxygen species (ROS) generation. Cell cycle analysis through flow cytometry using propidium iodide stain showed that P. acerifolium bark extract arrested A549 and PANC-1 cells in sub-G1 phase stated early apoptosis. These findings collectively point to the fact that P. acerifolium bark extract induced cell cytotoxicity in lung and pancreatic cancer cells by modulating mitochondrial-mediated ROS generation, and cell cycle checkpoints.

Tricycloalternarene Analogs from a Symbiotic Fungus Aspergillus sp. D and Their Antimicrobial and Cytotoxic Effects.

Bioassay-guided fractionation of the crude extract of fermentation broth of one symbiotic strain Aspergillus sp. D from the coastal plant Edgeworthia chrysantha Lindl. led to isolation of one new meroterpenoid, tricycloalternarene 14b (1), together with four known analogs (2-5), tricycloalternarenes 2b (2), 3a (3), 3b (4), and ACTG-toxin F (5). Their chemical structures were unambiguously established on the basis of NMR, mass spectrometry, and optical rotation data analysis, as well as by comparison with literature data. Biological assays indicated that compound 2 exhibited potent in vitro cytotoxicity against human lung adenocarcinoma A549 cell line with an IC50 value of 2.91 µM, and compound 5 had a moderate inhibitory effect on Candida albicans, with an MIC value of 15.63 µM. The results indicated that this symbiotic strain D is an important producer of tricycloalternarene derivatives, with potential therapeutic application in treatment of cancer and pathogen infection.

Southeast Asian Dipterocarp origin and diversification driven by Africa-India floristic interchange.

The evolution and diversification of ancient megathermal angiosperm lineages with Africa-India origins in Asian tropical forests is poorly understood because of the lack of reliable fossils. Our palaeobiogeographical analysis of pollen fossils from Africa and India combined with molecular data and fossil amber records suggest a tropical-African origin of Dipterocarpaceae during the mid-Cretaceous and its dispersal to India during the Late Maastrichtian and Paleocene, leading to range expansion of aseasonal dipterocarps on the Indian Plate. The India-Asia collision further facilitated the dispersal of dipterocarps from India to similar climatic zones in Southeast Asia, which supports their out-of-India migration. The dispersal pathway suggested for Dipterocarpaceae may provide a framework for an alternative biogeographic hypothesis for several megathermal angiosperm families that are presently widely distributed in Southeast Asia.

Potential Impacts of Translocation of Neonicotinoid Insecticides to Cotton (Gossypium hirsutum (Malvales: Malvaceae)) Extrafloral Nectar on Parasitoids.

Neonicotinoid seed treatments are frequently used in cotton (Gossypium hirsutum L. [Malvales: Malvaceae]) production to provide protection against early-season herbivory. However, there is little known about how these applications affect extrafloral nectar (EFN), an important food resource for arthropod natural enemies. Using enzyme-linked immunosorbent assays, we found that neonicotinoids were translocated to the EFN of clothianidin- and imidacloprid-treated, greenhouse-grown cotton plants at concentrations of 77.3 ± 17.3 and 122.6 ± 11.5 ppb, respectively. We did not find differences in the quantity of EFN produced by neonicotinoid-treated cotton plants compared to untreated controls, either constitutively or after mechanical damage. Metabolomic analysis of sugars and amino acids from treated and untreated plants did not detect differences in overall composition of EFN. In bioassays, female Cotesia marginiventris (Cresson) (Hymenoptera: Braconidae) parasitoid wasps that fed on EFN from untreated, clothianidin-treated, or imidacloprid-treated plants demonstrated no difference in mortality or parasitization success. We also conducted acute toxicity assays for C. marginiventris fed on honey spiked with clothianidin and imidacloprid and established LC50 values for male and female wasps. Although LC50 values were substantially higher than neonicotinoid concentrations detected in EFN, caution should be used when translating these results to the field where other stressors could alter the effects of neonicotinoids. Moreover, there are a wide range of possible sublethal impacts of neonicotinoids, none of which were explored here. Our results suggest that EFN is a potential route of exposure of neonicotinoids to beneficial insects and that further field-based studies are warranted.

Exploring the phytochemical profile of Cytinus hypocistis (L.) L. as a source of health-promoting biomolecules behind its in vitro bioactive and enzyme inhibitory properties.

Cytinus hypocistis whole plant and its three different parts (petals, stalks, and nectar) were chemically characterised and their biological properties evaluated. A total of 17 phenolic compounds were identified, being galloyl-bis-HHDP-glucose the most abundant. All the tested extracts showed high antioxidant capacity, with the petals exhibiting the most promising results both in the OxHLIA (IC50 = 0.279 ng/mL) and TBARS (IC50 = 0.342 ng/mL) assays. For the antidiabetic and anti-tyrosinase enzyme inhibitory assays, the stalk extract presented the lowest IC50 values, 0.039 mg/mL and 0.09 mg/mL, respectively. Regarding antibacterial activity, all tested extracts displayed broad-spectrum microbial inhibition against both Gram-positive and Gram-negative bacteria. Similarly, all extracts displayed effective anti-proliferation activity against four tested tumour cell lines (NCI-H460, HeLa, HepG2, and MCF-7), with no toxicity observed for a non-tumour cell line. Considering the anti-inflammatory activity, the petals showed the highest nitric oxide inhibition (IC50 = 127 µg/mL). These results point C. hypocistis as a promising source of health-promoting biomolecules.

Extract of Stellera Chamaejasme L. Inhibits the Progression of Hepatocellular Carcinoma by Regulating miR-134-5p and JAK1/STAT3 Pathway.

Background: Hepatocellular carcinoma (HCC) poses a growing threat to humans due to poor prognosis. Extract of stellera chamaejasme L. (ESC) is reported to inhibit metastasis of HCC. However, the underlying mechanism of ESC in regulating the progression of HCC needs to be further investigated. Methods: 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure cell proliferation. Flow cytometry was employed to check cell apoptosis. Transwell assay was conducted to assess the abilities of cell migration and invasion. The protein levels of proliferating cell nuclear antigen, cleaved caspase 3 (c-caspase 3), E-cadherin, janus kinase 1 (JAK1), signal transducer and activator of transcription 3 (STAT3), and phosphorylated STAT3 were detected by Western blot. The interaction between miR-134-5p and JAK1 was predicted by starBase, which was verified by the dual-luciferase reporter assay and RNA pull-down assay. The messenger RNA levels of miR-134-5p and JAK1 were determined by quantitative real-time polymerase chain reaction. Results: The results showed that the higher concentration or the longer time treatment of ESC led to the lower survival rate of HCC cells. Besides, ESC induced apoptosis and impeded migration and invasion of HCC cells. Moreover, downregulation of miR-134-5p inverted the effects of ESC-mediated repression on HCC progression. Further studies indicated that miR-134-5p targeted the 3'-untranslated region (3'UTR) of JAK1 and reversed JAK1-mediated impacts on HCC progression. Simultaneously, ESC inactivated JAK1/STAT3 pathway by regulating the expression of miR-134-5p. Conclusion: ESC suppressed HCC progression by upregulating the expression of miR-134-5p and blocking JAK1/STAT3 pathway.

Effects of DCM Leaf Extract of Gnidia glauca (Fresen) on Locomotor Activity, Anxiety, and Exploration-Like Behaviors in High-Fat Diet-Induced Obese Rats.

Obesity is the main component of metabolic syndromes involving distinct etiologies that target different underlying behavioral and physiological functions within the brain structures and neuronal circuits. An alteration in the neuronal circuitry stemming from abdominal or central obesity stimulates a cascade of changes in neurochemical signaling that directly or indirectly mediate spontaneously emitted behaviors such as locomotor activity patterns, anxiety, and exploration. Pharmacological agents available for the treatment of neurologic disorders have been associated with limited potency and intolerable adverse effects. These have necessitated the upsurge in the utilization of herbal prescriptions due to their affordability and easy accessibility and are firmly embedded within wider belief systems of many people. Gnidia glauca has been used in the management of many ailments including obesity and associated symptomatic complications. However, its upsurge in use has not been accompanied by empirical determination of these folkloric claims. The present study, therefore, is aimed at determining the modulatory effects of dichloromethane leaf extract of Gnidia glauca on locomotor activity, exploration, and anxiety-like behaviors in high-fat diet-induced obese rats in an open-field arena. Obesity was experimentally induced by feeding the rats with prepared high-fat diet and water ad libitum for 6 weeks. The in vivo antiobesity effects were determined by oral administration of G. glauca at dosage levels of 200, 250, and 300 mg/kg body weight in high-fat diet-induced obese rats from the 6th to 12th week. Phytochemical analysis was done using gas chromatography linked to mass spectroscopy. Results indicated that Gnidia glauca showed anxiolytic effects and significantly increased spontaneous locomotor activity and exploration-like behaviors in HFD-induced obese rats. The plant extract also contained phytocompounds that have been associated with amelioration of the main neurodegenerative mediators, viz., inflammation and oxidative stress. These findings provide "qualified leads" for the synthesis of new alternative therapeutic agents for the management of neurologic disorders. However, there is a need to conduct toxicity studies of Gnidia glauca to establish its safety profiles.