Antibacterial and antibiofilm activity of Lagotis brachystachya extract against extended-spectrum ß-lactamases-producing Escherichia coli from broiler chickens.

Lagotis brachystachya Maxim (L. brachystachya) is an herb widely used in traditional Tibetan medicine. In the present study, the antibacterial activity of L. brachystachya extract to extended-spectrum-lactamases (ESBLs)-producing E. coli was determined by Kirby-Bauer disc diffusion, minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) methods as well as time-kill curve assay. Meanwhile, the biofilm inhibition and eradication effects of L. brachystachya extract on the ESBLs-producing E. coli were evaluated by crystal violet staining, and further confirmed by confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The results indicated that L. brachystachya extract exhibited moderate antibacterial activity, with diameter of inhibition zones varying from 15.4 to 20.3 mm, and the MIC and MBC values were 6.25 to 25 mg/mL and 12.5 to 100 mg/mL, respectively. Time-kill curve showed that 4 × MIC level of L. brachystachya extract concentration of was able to kill 99.9% of ESBLs-producing E. coli after 16 h treatment. The biofilm inhibition rate and eradication rate for the ESBLs-producing E. coli were 35.66 to 79.91% and 22.18 to 56.21% at MIC level of extract concentration, respectively. CLSM images showed that the biofilm became thinner as the ESBLs-producing E. coli isolate exposed to L. brachystachya extract with a concentration-dependent manner from 1/4 × MIC to MIC compared with the control isolate. SEM images indicated that L. brachystachya extract at 1/2 × MIC and MIC levels could evidently inhibit the biofilm formation or eradicate the mature biofilms. The effect of L. brachystachya highlights its potential of antibacterial and antibiofilm activities against the ESBLs-producing E. coli.

In vitro phytochemical and anticancer activity of Misopates orontium L. and Dicliptera bupleuroides Nees.

In the present study phytochemical analysis and anticancer activity of Misopates orontium L. and Dicliptera bupleuroides Nees was carried out. Methanolic extracts of M. orontium and D. bupleuroides were selected for phytochemical analysis. The present analysis showed the presence of phytochemical such as carbohydrates, proteins, tannins, glycosides, alkaloids, saponins, phenols and flavonoids in M. orontium and D. bupleuroides. Anticancer assays including MTT, Alamar Blue (AB), Neutral Red (NR) and lactate dehydrogenase (LDH) were employed on whole herb methanolic extract and all other fractions of both plants to calculate the % age of cell viability and cell cytotoxicity. The percentage of cell viability was highly significant in all anticancer assays for all fractions. Therefore, ethyl acetate and aqueous fractions showed the excellent profile in evaluation of cytotoxicity in each assay. All above findings indicated that the whole herb of both selected plants have strong anticancer activity.

A review of the aromatic genus Adenosma: Geographical distribution, traditional uses, phytochemistry and biological activities.

ETHNOPHARMACOLOGICAL RELEVANCE: The strongly scented genus Adenosma R. Brown (Plantaginaceae) comprises between 26 and 29 species with mainly southeast Asian distributions. Several species are used traditionally, mostly in Asian countries, for medicinal purposes including the treatment of colds and tumors, as well as stomach, liver, and skin disorders. Some species are also used as insecticides and/or insect repellents against mosquitoes or fleas. AIM OF THE REVIEW: Although the potential health benefits of Adenosma spp. are not yet well-known or well-studied in modern medicine, the aim of the present review is to provide a critical appraisal of the current state of knowledge regarding the geographical distribution, traditional uses, phytochemistry, phytochemicals and biological properties of Adenosma spp. MATERIALS AND METHODS: Electronic databases (Web of Science, Science Direct, Google Scholar, Scifinder, Microsoft Academic, eFloras), Biodiversity Heritage Library (BHL), and the China National Knowledge Infrastructure (CNKI), were searched using the key words "Adenosma", "", "", "", "nhân tran", as well as the scientific names of the species, and a library search was also conducted for articles and books related to the subject published in English, Chinese or Vietnamese, as well as Ph.D. theses and M.Sc. dissertations published before April 2020. RESULTS: Adenosma spp. is traditionally used to treat gastrointestinal disorders, hepatitis, colds, and skin problems. Phenolic acids, flavonoids, and terpenoids constitute the main phytochemicals in these plants. Several evaluations based on bioassays have demonstrated biological activity for Adenosma spp., including antidiabetic, anticancer, and insecticidal activities; extracts and isolated compounds have also shown effective biological activity. However, current research has focused only on a few species, and on limited geographical regions, mainly in China and Vietnam. More and broader ethnopharmacological studies are therefore needed to provide further evidence of the health benefits of these plants. CONCLUSIONS: Adenosma spp. are plants rich in essential oils, particularly terpenoids, and the crude extracts have valuable bioactive properties. Certain lines of research based on cell lines and animal models show the potential value in different areas of health management. Further investigation into the traditional knowledge in southeast Asian and Pacific island regions, as well as the into the toxicity and identity of the bioactive compounds and their mechanisms of action is necessary.

Phytochemical Investigation and Antioxidant Activity of Globularia alypum L.

The Moroccan flora is rich in medicinal plants that are commonly used in folk medicine for the treatment of various diseases. The present study was designed to investigate the total phenolic and flavonoid contents, as well as the antioxidant properties of leaves extracts from Globularia alypum L. colected from the Taza region in northeast Morocco. Additionally, the individual phenolics and volatiles of the extracts were also evaluated. The organic extracts of this plant were obtained by Soxhlet extraction using two different solvents, namely ethyl acetate and chloroform. The antioxidant capacity of leaves extracts was measured using DPPH, ABTS and FRAP assays; the phenolic profile was determined by HPLC-DAD/ESI-MS analysis, whereas the volatile composition was elucidated by GC-MS. The ethyl acetate extract analysis showed a total of 20 phenolic compounds and the determination of phenolic contents showed a significant value of 56.5 ± 0.61 µg GAE/mg of extract in comparison with the chloroform extract (18.9 ± 0.48 µg GAE/mg of extract). Also, the determination of the flavonoid contents revealed that the ethyl acetate extract contained the highest value (30.2 ± 0.55 µg CE/mg of extract) in comparison with the chloroform extract (18.0 ± 0.36 µg CE/mg of extract). Concerning the antioxidant properties, interesting values were attained for the ethyl acetate extract which exhibited higher antioxidant activity, namely IC50 = 12.3 ± 3.83 µg/mL and IC50 = 37.0 ± 2.45 µg/mL for the DPPH and ABTS assays, respectively, whereas a value of 531.1 ± 17.08 (mg AAE/g DW) was obtained for the FRAP assay. Concerning the volatile profile, a total of 73 compounds were positively detected and among them n-hexadecanoic acid (13.5%) was the most abundant one. The results achieved confirm the important role of this plant as a source of natural antioxidants.

Plantaginis Semen polysaccharides ameliorate renal damage through regulating NLRP3 inflammasome in gouty nephropathy rats.

Gouty nephropathy (GN) is considered to be a prevalent renal disease and is an inflammatory event mainly induced by MSU crystals. Plantaginis Semen is a traditional Chinese herb that has been used in the treatment of gout, gouty arthritis and GN, but the mechanism and ingredients have been unclear. In this study, we explored and evaluated the preliminary structural characterizations of Plantaginis Semen polysaccharides (PSPs) and the activity of protecting against renal damage in GN rats. Three polysaccharide fractions, PSP-D, PSP-H and PSP-S, were sequentially extracted by different processes from the seed of Plantago asiatica L. The Fourier transform infrared spectral (FTIR) results showed that there were significant differences between PSP-S and the other two polysaccharides (PSP-D and PSP-H). PSP-D and PSP-H have pyrene monomers and linkages of ß-glycosides in their structures, and PSP-S has furanoside in the molecular structure. The scanning electron microscope (SEM) images of three polysaccharides showed that PSP-D has a smooth surface and a small curve, PSP-H is block-like and uneven in magnitude, whereas PSP-S is sea-tent-like and its surface is very distinct from the others. Main components and molar ratios are also different. Rats were randomly divided into six groups (n1/6 8 per group): the control group, model group, positive group, and three treatment groups (PSP-D, PSP-H and PSP-S). For all groups except the control group, rats were intragastrically administered the adenine suspension (50 mg kg-1 d-1) and fed with a high-yeast diet (15 g kg-1 d-1) for 28 days. On the 9th day, the control group and the model group were administered normal saline at the same time. Treatment groups were individually given corresponding drugs for 20 days. We found that PSPs could prevent renal damage, including decreasing the inflammatory response and regulating the (NOD)-like receptor protein 3 (NLRP3) protein in renal tissue. The underlying mechanism was related to NLRP3 inflammasome signal pathways, and it could take effect through the down-regulation of the protein expression levels of NLRP3, ASC and caspase-1 and inhibit the release of downstream inflammatory factors. PSPs are promising polysaccharides that could protect against renal injury through ameliorating renal inflammation in GN rats. Plantaginis Semen polysaccharides are potential functional food ingredients or pharmacological agents for treating GN in clinical practice.

Two new phenylpropanoid glycosides from Lagotis brachystachya Maxim and their Xanthione Oxidase inhibitions.

Two new phenylpropanoid glycosides lagotiside C and D, along with 11 known compounds were isolated from the whole plant of Lagotis brachystachya Maxim. The structures of lagotiside C and D was elucidated on the basis of spectroscopic data analysis. Moreover, all isolated components were evalued for the inhibition on Xanthione Oxidase (XOD) activity in vitro. Results indicated that all the compounds exhibited inhibitory effects on XOD with inhibition ratio in the range of 6.35%-83.69%, which suggested that Lagotis brachystachya could be served as an XOD inhibitor.

Composition and Larvicidal Activity of the Oil of Dizygostemon riparius (Plantaginaceae), a New Aromatic Species Occurring in Maranhão, Brazil.

Dizygostemon riparius (Plantaginaceae) is a new aromatic herbaceous species occurring in Maranhão State, Brazil. It is used as flavorings to remove domestic animal parasites and preventing mosquitoes. GC and GC/MS were used to analyze its essential oil, and a larvicidal bioassay was performed against Aedes albopictus larvae, a vector of arboviruses in Brazil. endo-Fenchyl acetate, endo-fenchol, (E)-caryophyllene, and caryophyllene oxide were the oil's primary constituents, totalizing 88.0 %. The two morphotypes of D. riparius, with purple and white flowers, did not present a significant difference in the oils' composition. From a chemotaxonomic point of view, D. riparius oil showed some similarities with other oils of Plantaginaceae, suggesting (E)-caryophyllene and caryophyllene oxide as possible chemical markers. The oil larvicidal action displayed the lowest and highest mortality percentage at 50 mg/L (2.0 %) and 600 mg/L (88.0 %). The fenchyl acetate and fenchol standards showed a lower and higher mortality percentage at a concentration of 300 mg/L (42.0 % and 26.0 %) and 900 mg/L (96.0 % and 98.0 %), respectively. The present study results with the D. riparius oil point to a new bioproduct with significant larvicidal activity.

Anticancer Activity of Novel Plant Extracts and Compounds from Adenosma bracteosum (Bonati) in Human Lung and Liver Cancer Cells.

Cancer is the second leading cause of death globally, and despite the advances in drug development, it is still necessary to develop new plant-derived medicines. Compared with using conventional chemical drugs to decrease the side effects induced by chemotherapy, natural herbal medicines have many advantages. The present study aimed to discover the potential cytotoxicity of ethanol extract and its derived fractions (chloroform, ethyl acetate, butanol, and aqueous) of Adenosma bracteosum Bonati. (A. bracteosum) on human large cell lung carcinoma (NCI-H460) and hepatocellular carcinoma (HepG2). Among these fractions, the chloroform showed significant activity in the inhibition of proliferation of both cancerous cells because of the presence of bioactive compounds including xanthomicrol, 5,4'-dihydroxy-6,7,8,3'-tetramethoxyflavone, and ursolic acid which were clearly revealed by nuclear magnetic resonance spectroscopy (1H-NMR, 13C-NMR, Heteronuclear Multiple Bond Coherence, and Heteronuclear Single Quantum Coherence Spectroscopy) analyses. According to the radical scavenging capacity, the 5,4'-dihydroxy-6,7,8,3'-tetramethoxyflavone compound (AB2) exhibited the highest anticancer activity on both NCI-H460 and HepG2 with IC50 values of 4.57 ± 0.32 and 5.67 ± 0.09 µg/mL respectively, followed by the ursolic acid with the lower percent inhibition at 13.05 ± 0.55 and 10.00 ± 0.16 µg/mL, respectively (p < 0.05). Remarkably, the AB2 compound induced to significant increase in the production of reactive oxygen species accompanied by attenuation of mitochondrial membrane potential, thus inducing the activation of caspase-3 activity in both human lung and liver cancer cells. These results suggest that A. bracteosum is a promising source of useful natural products and AB2 offers opportunities to develop the novel anticancer drugs.

Essential oil from leaves of Conobea scoparioides (Cham. & Schltdl.) Benth. (Plantaginaceae) causes cell death in HepG2 cells and inhibits tumor development in a xenograft model.

Conobea scoparioides (Cham. & Schltdl.) Benth. (syn. Sphaerotheca scoparioides Cham. & Schldtl.) (Plantaginaceae), popularly known as "pataqueira", "vassourinha-do-brejo" and/or "hierba-de-sapo", is a popular medicinal plant used to treat leishmaniasis, pain and beriberi. In addition, inhibition of cell adhesion, antioxidant, cytotoxic and leishmanicidal activities of compounds or fractions of C. scoparioides have been reported. In the present work, chemical constituents and in vitro and in vivo anti-liver cancer potential of essential oil (EO) from leaves of C. scoparioides were investigated using human hepatocellular carcinoma HepG2 cells as a cell model. EO was obtained by hydrodistillation using a Clevenger-type apparatus and characterized by GC-MS and GC-FID. The in vitro cytotoxic effect was evaluated on three human cancer cell lines (MCF-7, HepG2 and HCT116) and one human non-cancerous cell line (MRC-5) using the Alamar blue assay. Phosphatidylserine externalization and cell cycle distribution were quantified in HepG2 cells by flow cytometry after 48 h incubation. The effectiveness of EO in anti-liver cancer model was studied with HepG2 cells grafted on C.B. 17 SCID mice. The main constituents of EO were thymol methyl ether (62 %), thymol (16 %) and α-phellandrene (14 %). EO displayed an in vitro cytotoxic effect against all human cancer cell lines and caused externalization of phosphatidylserine and DNA fragmentation in HepG2 cells, suggesting induction of apoptotic-like cell death. In vivo tumor mass inhibition of 36.7 and 55.8 % was observed for treatment with EO at doses of 40 and 80 mg/kg, respectively. These results indicate in vitro and in vivo anti-liver cancer potential of EO from leaves of C. scoparioides.

Guided isolation of new iridoid glucosides from Anarrhinum pubescens by high-performance thin-layer chromatography-acetylcholinesterase assay.

Plants are an important source of natural iridoids. This study demonstrates for the first time the acetylcholinesterase (AChE) inhibitory activity of iridoids belonging to the class of antirrhinosides. As iridoids distinguish the chemical composition of most species of the Plantaginaceae family, the active AChE inhibitors were investigated in the hydro-alcoholic extract of Anarrhinum pubescens Fresen. High-performance thin-layer chromatography (HPTLC) in combination with the AChE inhibition assay is a time and material saving methodology, and thus was employed to directly point to the individual enzyme inhibitors occurring in the plant. The effect-directed screening successfully discovered three active metabolites. These were characterized as antirrhinoside-derived iridoids. Two of these are here reported as newly isolated natural compounds. Identification of the two new metabolites was based on analysis of their collected spectroscopic data (HRMS, 1D and 2D NMR). Their structures were elucidated to be 6-O-, 6'-O-di-trans-cinnamoyl-antirrhinoside (1) and 5-O-, 6-O-difoliamenthoyl-antirrhinoside (3), while the previously known compound 6-O-foliamenthoyl-(6'-O-cinnamoyl)-antirrhinoside (2) was assigned by extensive analysis of its HRMS and HRMS/MS data. The activity of the isolated compounds was referred to the known AChE inhibitor rivastigmine, i.e. their activity were calculated and expressed as values equivalently to rivastigmine. This neuroprotective potential of iridoids mediated through AChE inhibition promote them to compete as natural curatives for neurodegenerative disorders like Alzheimer's disease.

Neopicrorhiza scrophulariiflora (Pennell) Hong: A comprehensive review of its traditional uses, phytochemistry, pharmacology and safety.

ETHNOPHARMACOLOGICAL RELEVANCE: Neopicrorhiza scrophulariiflora (Pennell) Hong is important medicinal plant that is native to the eastern Himalayas and Hengduan mountains in China. It is also distributed in Nepal, north east India, Bhutan and northern Myanmar. Plant parts are traditionally used against different kinds of diseases and various compounds present in different plant parts are also effective against many diseases. Thus, N. scrophulariiflora has a high potential to maintain human health. AIM OF THE REVIEW: Although N. scrophulariiflora is very important and widely studied plant species but there is no comprehensive up-to-date review of published and unpublished literature. So, in the present article we have compiled and critically commented on the botanical characteristics, traditional uses, plant growth and cultivation, micropropagation, conservation status, secondary metabolites, pharmacology and toxicity of the plant. MATERIALS AND METHODS: Extensive literature searches both electronic online databases (Google Scholar, Scopus, Springer Link, Web of Science, ScienceDirect, ResearchGate, PubMed, ChemSpider, USPTO, Google patents and Espacenet) and library visits in Nepal were carried out to collect the literature on information published prior to April 2019. RESULTS: N. scrophulariiflora was traditionally used for 82 ailments/diseases. There are 124 major phytochemicals extracted from the plant. Several compounds are effective in bioactivity. Pharmacologically, the plant is proved to be anti-atherosclerotic, antidiabetic and anti-inflammatory in-vivo studies, and antimicrobial, antimalarial, antioxidative, hepatoprotective, immunomodulatory and nerve growth factor potentiating from in-vitro studies. Renal improvement activities were confirmed from both in-vivo and in-vitro studies. Toxicological tests and a single clinical trial in human beings have supported the notion that the plant is not poisonous but beneficial for curing wide ranges of diseases. CONCLUSION: N. scrophulariiflora is valuable medicinal plant that can serve as promising source of non-harmful and potential medicinal herbal remedies for human beings.

Actualities in the phytochemical research on selected terpenes.

A short review of our recent research on the essential oil phytochemical composition of Petasites albus (L.) Gaertn. and Petasites hybridus (L.) G. Gaertn., B. Mey. & Scherb. (Asteraceae) as well as on the oils of Globularia cordifolia L., Globularia meridionalis (Podp.) O. Schwarz and Globularia punctata Lapeyr. (Plantaginaceae) is presented. All essential oils contained a variety of oxygenated sesquiterpenes among their major constituents, including a bakkane type sesquiterpene fukinanolid (bakkenolide A). The paper is focused on: i) a short overview of the abundance of major terpenes in the essential oils of Petasites and Globularia species from Croatia; ii) possible biosynthetic pathways of major identified sesquiterpenes; and iii) biological activities (literature data) of major sesquiterpenes from Petasites and Globularia species.

Picroside II Isolated from Pseudolysimachion rotundum var. subintegrum Inhibits Glucocorticoid Refractory Serum Amyloid A (SAA) Expression and SAA-induced IL-33 Secretion.

Chronic obstructive pulmonary disease (COPD) is a major inflammatory lung disease characterized by irreversible and progressive airflow obstruction. Although corticosteroids are often used to reduce inflammation, steroid therapies are insufficient in patients with refractory COPD. Both serum amyloid A (SAA) and IL-33 have been implicated in the pathology of steroid-resistant lung inflammation. Picroside II isolated from Pseudolysimachion rotundum var. subintegrum (Plantaginaceae) is a major bioactive component of YPL-001, which has completed phase-2a clinical trials in chronic obstructive pulmonary disease patients. In this study, we investigated whether picroside II is effective in treating steroid refractory lung inflammation via the inhibition of the SAA-IL-33 axis. Picroside II inhibited LPS-induced SAA1 expression in human monocytes, which are resistant to steroids. SAA induced the secretion of IL-33 without involving cell necrosis. Picroside II, but not dexamethasone effectively inhibited SAA-induced IL-33 expression and secretion. The inhibitory effect by picroside II was mediated by suppressing the mitogen-activated protein kinase (MAPK) p38, ERK1/2, and nuclear factor-κB pathways. Our results suggest that picroside II negatively modulates the SAA-IL-33 axis that has been implicated in steroid-resistant lung inflammation. These findings provide valuable information for the development of picroside II as an alternative therapeutic agent against steroid refractory lung inflammation in COPD.

Antiangiogenic Activity of Compounds Isolated from Anarrhinum pedatum.

Ten new iridoid glycosides (1-10) and two new monoterpenoids (11 and 12), together with nine known compounds (13-21), were isolated from the n-butanol extract of the aerial parts of Anarrhinum pedatum. The structural characterization of all compounds was performed by spectroscopic analysis, including 1D and 2D NMR and HRESIMS experiments. The isolates were assayed for their antiangiogenic activity by two in vivo models, using zebrafish embryos and chicken chorioallantoic membranes (CAMs). The results showed that among the new compounds 6'- O-menthiafoloylmussaenosidic acid-11-(5- O-ß-d-fructopyranosyl) ester (9) exhibited the most potent antiangiogenic activity in both the zebrafish embryos and CAM assays, reducing the growth of blood vessels. Antiangiogenic effects were also observed for the known compounds 6- O-nerol-8-oyl-antirrinoside (13), antirrinoside (14), 6- O- trans- and cis- p-coumaroyl antirrinoside (15), and (6 S)-2 E-2,6-dimethyl-6-hydroxyocta-2,7-dienoic acid ß-glucopyranosyl ester (18).

Microwave irradiation followed by zinc oxide based dispersive solid-phase extraction coupled with HPLC for simultaneous extraction and determination of flavonoids in Veronicastrum latifolium (Hemsl.) Yamazaki.

In this study, a sample preparation method involving microwave irradiation followed by dispersive solid-phase extraction was used to extract and separate flavonoids from Veronicastrum latifolium (Hemsl.) Yamazaki. Ethyl lactate was selected as the extraction solvent during microwave irradiation. Zinc oxide and ethanol were used as the adsorbents and the eluent, respectively, in dispersive solid-phase extraction. Several experimental parameters affected the extraction performance, such as the extraction solvent and method, the microwave irradiation time, temperature, and solid-to-liquid ratio, the type of adsorbent, the amount of sample solution, the adsorption time and method, and the type and volume of the eluent, and were investigated by single-factor and Box-Behnken design experiments in the pretreatment process. The recommended method was applied to extract flavonoids in Veronicastrum latifolium (Hemsl.) Yamazaki. The enriched flavonoids were analyzed for the first time by high-performance liquid chromatography, and seven flavonoids-vitexin, luteolin, wogonoside, apigenin, chrysoeriol, acacetin, and pectolinarigenin-were identified. Additionally, the method exhibited good linearity (r2 ≥ 0.9992) for flavonoids under the optimum conditions. Moreover, the relative standard deviation of the intraday and interday precision was less than 3.97%. The mean recoveries for flavonoids were more than 98.10%. This work is the first to report a method to extract, separate, and analyze flavonoids from Veronicastrum latifolium (Hemsl.) Yamazaki. Graphical Abstract Microwave irradiation followed by ZnO-based dispersive solid-phase extraction for the pretreatment of Veronicastrum latifolium (Hemsl.) Yamazaki samples, and high-performance liquid chromatography (HPLC) for analysis of enriched flavonoids.

Plant-derived verbascoside and isoverbascoside regulate Toll-like receptor 2 and 4-driven neutrophils priming and activation.

BACKGROUND: Neutrophils have a short live in circulation and accelerate greatly local immune responses via increased granulopoiesis and migration at high numbers to infected or inflamed tissue. HYPOTHESIS: Since neutrophils produce a variety of factors with destructive and pro-inflammatory potential the regulation of their homeostasis and functions might be eventually beneficial in inflammation-related pathological conditions. Herein we investigated the effect of natural-derived verbascoside (Verb) and its positional isomer isoverbascoside (IsoVerb) on neutrophil functions. METHODS: We used purified murine bone marrow (BM) neutrophils to study cell responsiveness to priming or activation via Toll-like receptors (TLRs) 2 and 4. The expression of CD11b, chemokine (CXC motif) receptor 2 (CXCR2), the intracellular level of phosphorylated p38 mitogen-activated protein kinase (MAPK) and tumor necrosis factor (TNF)-α in neutrophils were determined by flow cytometry while the release of macrophage inflammatory protein (MIP)-2 in culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: We found that Verb appeared less powerful inhibitor of TLR2 and TLR4-mediated apoptosis than IsoVerb. However at concentrations below 16 µM and in LPS priming conditions Verb was more selective inhibitor of CD11b and CXCR2 expression than IsoVerb. Both compounds showed similar activity on integrin/chemokine receptor expression when neutrophils were stimulated with ZY or were activated with LPS. Verb sustained CXCR2 expression and turnover via regulation of the cell responsiveness to its ligand KC (CXCL1) and via the release of MIP-2 (CXCL2). Both Verb and IsoVerb increased TNF-α production and inhibited p38 phosphorylation in TNF-α+ cells. We fail to discriminate sharply between Verb's and IsoVerb's efficacy when studying p38 phosphorylation in LPS stimulated neutrophils. The multi-parametric analysis provides critical insight on the range of on-target effects of Verb and IsoVerb. CONCLUSION: The strength and selectivity of Verb and IsoVerb depended on the degree of activation and functional state of neutrophils, and both compounds are with potential to affect neutrophil-related pathologies/conditions in heterogenic populations.

Bioassay-guided isolation of active compounds from Adenosma buchneroides essential oil as mosquito repellent against Aedes albopictus.

ETHNOPHARMACOLOGICAL RELEVANCE: A folk herb Adenosma buchneroides found in the previous ethnobotanical investigation plays an important role as an insect repellent among the Aini people in southwest of China, but the active compounds responsible for repellent activity of the plant have not yet been investigated. AIM OF THE STUDY: The main purpose of the study is to identify the active components of the essential oil which responsible for its repellent activity against Aedes albopictus to support the usage of the plant as mosquito repellent by Aini people. In addition, to supply a class of potential alternatives characterized carvacrol analogues to develop natural repellent products. MATERIAL AND METHODS: The essential oil from aerial part of Ad. buchneroides was extracted by hydrodistillation. A systematic bioassay-guided isolation of repellent compounds from the essential oil was conducted through chromatographic fractionation combined with in-cage mosquito repellent bioassay. The identification of the essential oil components was accomplished by GC-MS and GC-FID techniques. The structural elucidation of compounds was performed on the basis of IR, HR-ESI-MS and NMR. Larvicidal activity and cytotoxicity of all repellent compounds also tested by larval bioassays and MTS assays, respectively. Structure-activity relationship (SAR) of carvacrol analogues was investigated by in-cage mosquito repellent bioassay. RESULTS: The essential oil of the plant showed strong mosquito repellent activity with minimum effective dosage (MED) of 0.019 ±â€¯0.007 mg/cm2, compared to reference standard N,N-diethyl-3-methylbenzamide (DEET) (0.031 ±â€¯0.014 mg/cm2). 26 compounds representing 97.8% of the essential oil were identified. Carvacrol, carvacrol methyl ether and a new fragrant compound, adenosmin A (1) were found to be repellent compounds by systematic bioassay-guided isolation, with MEDs in the range of 0.011-0.125 mg/cm2. An investigation on SAR of carvacrol analogues led to the discover of three analogues with further lower MEDs (0.002-0.009 mg/cm2) than that of DEET, and other three compounds with similar MEDs (0.029-0.039 mg/cm2) to that of DEET. Carvacrol (LD50 of 24.8 ppm) was the best larvicide among tested repellent compounds. The essential oil and repellent compounds against seven mammalian cell lines revealed low or no cytotoxicity. CONCLUSIONS: Scientific evidences reported here validate the plant's traditional use as insect repellent and imply promising application of the essential oil and carvacrol analogues as natural mosquito repellents.

New iridoid glycosides from Anarrhinum pubescens.

Six metabolites (1-6) were isolated from the aerial parts of Anarrhinum pubescens Fresen. (Plantaginaceae) growing in Saint Catherine region in Egypt; two of them (1 and 4) are here reported to be newly identified naturally occurring iridoids. The isolated metabolites were identified as 6-O-foliamenthoyl-(6'-O-cinnamoyl)-antirrhinoside (1), 6'-O-cinnamoyl-antirrhinoside (2), the iridoid dimer, pubescensoside (4), antirrhinoside (5), 10-hydroxy-antirrhinoside (6), and the flavonoid, diosmin (3). Identification of the new metabolites was based on analysis of their collected spectroscopic data (NMR and HR-ESI-MS). Furthermore, compounds (1, 4, and 5) were subjected to cytotoxic testing against the human lung carcinoma cell line (A-549); compound 4 showed better cytotoxic activity as indicated by the obtained (IC50).

Sulfated polysaccharide isolated from Globularia alypum L.: Structural characterization, in vivo and in vitro anticoagulant activity, and toxicological profile.

A sulfated polysaccharide from Globularia alypum L. (GASP) was extracted with a yield of 14.2%. GASP is composed mostly of sulfate and total sugars (13.29% and 71.56%, respectively) with small amount of proteins and lipids. The chemical and structural characterization was studied by Infra-Red spectroscopic and gas chromatography-mass spectrometry (GC-MS). GASP composed of eight carbohydrates where galactose, glucose, and mannose are the major compounds (33.47%, 26.71% and 18.21%, respectively). The in vitro and in vivo anticoagulant activities in rats were tested using the standard coagulation assays activated partial thromboplastin time (aPTT), prothrombine time (TT) and thrombin time (PT) tests. Both doses of GASP (200 and 500 mg/kg b.w) displayed a significant in vitro (1.22 and 1.33-fold, 1.17 and 1.27-fold, and 1.21 and 1.26-fold, respectively) and in vivo (1.47 and 2.52-fold; 1.20 and 1.43-fold; 1.21 and 1.40-fold, respectively) compared with the control. Toxicity studies on liver performed by the catalytic activity of transaminases in plasma, oxidative stress markers and hepatic morphological changes indicated that GASP at both doses are not toxics. The important pharmacological and toxicological profile of GASP revealed that this compound may be used as a novel and effective drug.

Chemical fingerprint and bioactivity evaluation of Globularia orientalis L. and Globularia trichosantha Fisch. & C. A. Mey. using non-targeted HPLC-ESI-QTOF-MS approach.

INTRODUCTION: In the quest for new sources of biologically-active compounds, the chemical, and biological profiles of two Globularia species (G. trichosantha Fisch. & C. A. Mey and G. orientalis L.) were investigated. METHODOLOGY: Chemical profiles were evaluated by high-performance liquid chromatography coupled to electrospray ionisation and quadrupole time-of-flight mass spectrometry (HPLC-ESI-QTOF-MS), as well as by their total phenolic, flavonoids, and phenolic acids contents. The antioxidant abilities of the investigated extracts were done using different assays including free radical scavenging [1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS)], reducing power (cupric reducing antioxidant capacity and ferric reducing antioxidant power), phosphomolybdenum, and metal chelating. Inhibitory potential against key enzymes involved in neurodegenerative diseases (cholinesterases; AChE, and BChE), diabetes (α-glucosidase and α-amylase), hyperpigmentation (tyrosinase) and obesity (pancreatic lipase) were evaluated. RESULTS: Globularia trichosantha and G. orientalis extracts showed remarkable antioxidant properties, with the water extracts being a better source of antioxidant compounds. Both species showed remarkable inhibitory effects against the target enzymes. However, for both species, only the acetyl acetate and methanolic extracts were potent against cholinesterases and lipase. CONCLUSION: HPLC-ESI-QTOF-MS analysis revealed the presence of 107 compounds from G. trichosantha and G. orientalis, among which, 43 compounds have been preliminarily characterised for the first time from the Globulariaceae family. To date, this study can be considered as the most comprehensive research focused on the characterisation of G. trichosantha and G. orientalis. Results amassed from this study tend to show that these plants represent a rich source of biologically active compounds which can be further explored and validated for their therapeutic potential.