VAMP726 from maize and Arabidopsis confers pollen resistance to heat and UV radiation by influencing lignin content of sporopollenin.

Sporopollenin in the pollen cell wall protects male gametophytes from stresses. Phenylpropanoid derivatives, including guaiacyl (G) lignin units, are known to be structural components of sporopollenin, but the exact composition of sporopollenin remains to be fully resolved. We analyzed the phenylpropanoid derivatives in sporopollenin from maize and Arabidopsis by thioacidolysis coupled with nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry (GC-MS). The NMR and GC-MS results confirmed the presence of p-hydroxyphenyl (H), G, and syringyl (S) lignin units in sporopollenin from maize and Arabidopsis. Strikingly, H units account for the majority of lignin monomers in sporopollenin from these species. We next performed a genome-wide association study to explore the genetic basis of maize sporopollenin composition and identified a vesicle-associated membrane protein (ZmVAMP726) that is strongly associated with lignin monomer composition of maize sporopollenin. Genetic manipulation of VAMP726 affected not only lignin monomer composition in sporopollenin but also pollen resistance to heat and UV radiation in maize and Arabidopsis, indicating that VAMP726 is functionally conserved in monocot and dicot plants. Our work provides new insight into the lignin monomers that serve as structural components of sporopollenin and characterizes VAMP726, which affects sporopollenin composition and stress resistance in pollen.

The Immunologic Profiles of Kawasaki Disease Triggered by Mycoplasma pneumoniae Infection.

OBJECTIVE: We compared the immunologic characteristics of mycoplasma pneumoniae-triggered Kawasaki disease (MP-KD) with Kawasaki disease (KD) not associated with mycoplasma pneumoniae (MP), with mycoplasma pneumoniae-triggered Henoch-Schönlein purpura (MP-HSP), and with healthy controls. METHODS: Complement levels, cellular and humoral immunity were assessed in KD, in MP-KD, in MP-HSP, and in healthy children. RESULTS: Of 622 children with KD, 74 had MP-KD. Complement C3 and CD4/CD8 ratio were significantly increased in MP-KD compared to KD. C3, C4, and the ratio of CD4/CD8 in the MP-KD group were higher than those in the MP-HSP group. IgA and CD56 were lower in the MP-KD group than the MP-HSP group. CONCLUSIONS: Both C3 and polyclonal CD4+ T lymphocytes may be activated in the patients with MP-KD.

Development of a reliable UHPLC-MS/MS method for simultaneous determination of zearalenone and zearalenone-14-glucoside in various feed products.

A reliable ultra-high-performance liquid chromatography-tandem mass spectrometry method (UHPLC-MS/MS) was developed for the simultaneous determination of two mycotoxins, that is, zearalenone (ZEN) and zearalenone-14-glucoside (ZEN-14G) in formula feed, concentrated feed, and premixed feed products. An improved sample pretreatment was achieved with the hydrophilic-lipophilic balance (HLB) cartridges efficiently removing the impurities and enriching the target analytes in different feeds. The critical parameters affecting the performance of the solid-phase extraction (SPE) procedure were carefully optimized, and 20% acetonitrile in water as the loading solution, 50% methanol in water as the washing solvent, and 5 ml of methanol as the elution solvent yielded the optimal purification efficiencies. The established method was thoroughly validated in terms of linearity (R 2 ≥ 0.999), sensitivity (limit of quantification in the range of 0.50-5.00 µg kg-1), recovery (89.35 ± 2.67% to 110.93 ± 1.56%), and precision (RSD, 3.00-14.20%), and it was then successfully applied to investigate a total of 60 feed samples. Among them, 50 samples were found to be contaminated with ZEN (an incidence of 83.3%) at levels ranging from 0.63 to 615.24 µg kg-1, whereas 22 samples were contaminated with ZEN-14G (an incidence of 36.7%) in the range of 0.89-15.31 µg kg-1. The developed method proved to be a specific and reliable tool for intensive monitoring of ZEN and ZEN-14G in complex feed matrices.

Genotyping-by-Sequencing of Gossypium hirsutum Races and Cultivars Uncovers Novel Patterns of Genetic Relationships and Domestication Footprints.

Determining the genetic rearrangement and domestication footprints in Gossypium hirsutum cultivars and primitive race genotypes are essential for effective gene conservation efforts and the development of advanced breeding molecular markers for marker-assisted breeding. In this study, 94 accessions representing the 7 primitive races of G hirsutum, along with 9 G hirsutum and 12 Gossypium barbadense cultivated accessions were evaluated. The genotyping-by-sequencing (GBS) approach was employed and 146 558 single nucleotide polymorphisms (SNP) were generated. Distinct SNP signatures were identified through the combination of selection scans and association analyses. Phylogenetic analyses were also conducted, and we concluded that the Latifolium, Richmondi, and Marie-Galante race accessions were more genetically related to the G hirsutum cultivars and tend to cluster together. Fifty-four outlier SNP loci were identified by selection-scan analysis, and 3 SNPs were located in genes related to the processes of plant responding to stress conditions and confirmed through further genome-wide signals of marker-phenotype association analysis, which indicate a clear selection signature for such trait. These results identified useful candidate gene locus for cotton breeding programs.

An effective identification and quantification method for Ginkgo biloba flavonol glycosides with targeted evaluation of adulterated products.

BACKGROUND: Ginkgo biloba L. (Ginkgoaceae) leaf extract is one of the most popular herbal products on the market, as it contains flavone glycosides (≥ 24%) and terpene lactones (≥ 6%), which are proposed to have significant physiological effects. Unfortunately, the challenging financial climate has resulted in a natural health product market containing adulterated ginkgo products. PURPOSE: 42 ginkgo samples were analyzed to establish an HPLC profile for authentic ginkgo and common ginkgo adulterants, and to develop a method capable of easily detecting adulteration in ginkgo commercial products. METHOD: In this study an efficient and targeted HPLC analysis method was established that is capable of distinguishing flavonol glycosides and aglycones simultaneously for the evaluation of ginkgo powdered extracts (PEs) and finished products in a single, 13 min run. Thirteen ginkgo leaf samples, fifteen standardized powdered extracts, and fourteen commercially available ginkgo products have been analyzed using this new HPLC method. Chromatograms were compared to six standard reference materials: one flavonol glycoside (rutin), three aglycones (quercetin, kaempferol and isorhamnetin), and two isoflavones (genestin and genistein). The quantitative chromatographic data was interpreted by principal component analysis (PCA), which assisted in the detection of unexpected chromatographic features in various adulterated botanical products. RESULTS: Only three of the commercially available ginkgo finished products tested in this study were determined to be authentic, with flavonol glycoside rutin, and aglycones quercetin, kaempferol, and isorhamnetin found to be common adulterants in the ginkgo powdered extract and finished product samples. CONCLUSION: Despite evidence of adulteration in most of the samples, each of the samples discussed herein met most of the current pharmacopeial standards. It is therefore critical that a preliminary evaluation be utilized to detect adulteration in commercial ginkgo products, prior to the acid hydrolysis procedure utilized in the current testing methods.

Anti-tumor and anti-angiogenic effects of Macrothelypteris viridifrons and its constituents by HPLC-DAD/MS analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Macrothelypteris viridifrons is widely distributed in south of China and has been used as folk medicine to treat cancer, hydropsy, and traumatic bleeding. AIM OF THE STUDY: To investigate the chemical constituents and the anti-tumor and anti-angiogenic effects of Macrothelypteris viridifrons. MATERIALS AND METHODS: An HPLC-DAD/MS technique was used to determine the flavonoid profile of Macrothelypteris viridifrons. The anti-tumor effect of Macrothelypteris viridifrons was evaluated by in vivo mice bearing H22 hepatoma cells transplantation tumor model. And the anti-angiogenic activity was investigated by measuring the effects on the in vitro proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs). Furthermore, the in vivo zebrafish model was applied to evaluate the anti-angiogenic effect of Macrothelypteris viridifrons. RESULTS: 18 flavonoids were identified from Macrothelypteris viridifrons. Administration of Macrothelypteris viridifrons significantly inhibited the tumor growth and the expression of vascular endothelial growth factor (VEGF) and CD34. Meanwhile, Macrothelypteris viridifrons showed significant inhibition on proliferation, migration and tube formation of HUVECs in vitro and the intersegmental vessels formation in zebrafish model. CONCLUSIONS: Macrothelypteris viridifrons showed significant anti-tumor and anti-angiogenic effects and might be developed as a novel anti-tumor drug.

Nephroprotective activity of Macrothelypteris oligophlebia rhizomes ethanol extract.

CONTEXT: Macrothelypteris oligophlebia (Bak.) Ching (Thelypteridaceae) is a Chinese herbal medicine used traditionally for the treatment of diseases such as edema, boils, burns, and roundworms. However, research about the nephroprotective potential of this plant is not available. OBJECTIVE: Present study was designed to evaluate the protective effect of ethanol extract of M. oligophlebia rhizomes (EMO) on gentamicin (GM)-induced nephrotoxicity. MATERIALS AND METHODS: Rats were intraperitoneal (i.p.) injected with GM (100 mg/kg) to induce nephrotoxicity and simultaneously EMO (250 and 500 mg/kg) was orally given to GM-treated rats for 8 days. Blood urea nitrogen (BUN), serum creatinine (Cr), malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were evaluated in renal tissues. Histopathological analysis was used for evaluation of the renal damage. RESULTS: Administration with GM-induced renal dysfunction in rats. Pre-treatment with EMO (500 mg/kg) significantly decreased the levels of BUN, Cr, MDA and NO (decreased BUN from 12.71 ± 1.28 to 7.19 ± 0.23 mmol/l, Cr from 39.77 ± 5.34 to 19.17 ± 0.90 µmol/l, MDA from 5.60 ± 0.37 to 2.63 ± 0.24 nmol/ml, and NO from 868.17 ± 22.67 to 589.51 ± 8.83 µmol/ml), and also restored the activities of renal antioxidant enzymes (SOD, CAT, and GSH-Px) (restored SOD from 1.59 ± 0.17 to 2.94 ± 0.13 U/mg protein, CAT from 3.22 ± 0.34 to 10.57 ± 0.27 U/mg protein, and GSH-Px from 9.11 ± 1.29 to 20.72 ± 1.83 U/mg protein). DISCUSSION AND CONCLUSION: Our results suggest that the rhizomes of M. oligophlebia potentially have a protective role in renal tissue against oxidative stress in acute renal failure.

Characterisation of phenols and antioxidant and hypolipidaemic activities of Lethariella cladonioides.

BACKGROUND: This study aimed to investigate the antioxidant and hypolipidaemic activities of an ethanol extract of Lethariella cladonioides (Nyl.) Krog (EE) and to characterise its chemical constituents. RESULTS: Nine phenols were identified as canarione, thamnolic acid, squamatic acid, vermicularin, norstictic acid, baeomycesis acid, lecanoric acid, barbatinic acid and usnic acid from analysis of EE by using high-performance liquid chromatography with a diode array detector-mass spectrometry. In antioxidant analysis in vitro, the highest scavenging rate of EEs on the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, superoxide anion, hydroxyl radicals and hydrogen peroxide was 81.55 ± 1.95%, 81.84 ± 4.00%, 74.28 ± 3.71% and 74.28 ± 3.71%, respectively. Meanwhile, after administration of EE for 6 weeks in high fat/cholesterol diet mice, the most significant reduction in levels of serum triglyceride, serum total cholesterol, serum low density lipoprotein cholesterol and liver malondialdehyde were 24%, 20%, 15% and 35%, respectively. The most significant increase in levels of serum high density lipoprotein cholesterol and liver superoxide dismutase was 35% and 88%, respectively. CONCLUSION: L. cladonioides possesses strong antioxidant and hypolipidaemic activities.

A novel non-aromatic B-ring flavonoid: isolation, structure elucidation and its induction of apoptosis in human colon HT-29 tumor cell via the reactive oxygen species-mitochondrial dysfunction and MAPK activation.

The aim of the present study was to elucidate the chemical structure of a novel non-aromatic B-ring flavonoid (DHEC) isolated from Macrothelypteris viridifrons and to evaluate its putative molecular mechanism of action on induction of apoptosis in human colon HT-29 cancer cell. On the basis of MS, UV, IR, 1D and 2D NMR data, DHEC was identified as 2-(cis-1, 2-dihydroxy-4-oxo-cyclohex-5-enyl)-5-hydroxy-7-ethoxy-chromone. In addition, the cytotoxicity of DHEC and its effect on induction of apoptosis were confirmed by several assays. After treatment of HT-29 cell with DHEC, we observed the accumulation of intracellular reactive oxygen species, the loss of mitochondrial membrane potential, the alteration of expression of the Bcl-2 family members, the releasing of cytochrome c, the cleavage of poly (ADP-ribose) polymerase (PARP), and the activation of caspase-3, -8, and -9. Further analysis showed that the mitogen-activated protein kinase (MAPK) related proteins were stimulated by treatment with DHEC. These results suggest that DHEC exhibits potential anti-cancer activity in HT-29 cell through induction of apoptosis, which may highly be associated with reactive oxygen species-mitochondrial dysfunction as well as activation of MAPK signaling pathway.

Topical anti-inflammatory and analgesic activity of kirenol isolated from Siegesbeckia orientalis.

ETHNOPHARMACOLOGICAL RELEVANCE: Siegesbeckia orientalis has been traditionally used as a topical anti-inflammatory and analgesic agent. AIMS OF THE STUDY: Current study was designed to explore the topical anti-inflammatory and analgesic effects of a constituent isolated from Siegesbeckia orientalis (Compositae), in order to validate its folk use. MATERIALS AND METHODS: Kirenol was isolated from ethanolic extract of Siegesbeckia orientalis. Several topical formulations containing kirenol were investigated for anti-inflammatory and analgesic activities in rat. The effects were studied using carrageenan-induced rat acute inflammation model, complete Freund's adjuvant (CFA)-induced chronic inflammation and formalin test in rats. Piroxicam gel and methyl salicylate ointment were studied as positive control for anti-inflammatory and analgesic activity, respectively. RESULTS: The anti-inflammatory effect of kirenol 0.4-0.5% (w/w) was similar to the effect of piroxicam gel 4h after carrageenan injection. The analgesic activity of topical preparation with more than 0.4% (w/w) was observed in the late phase. These effects may be due, at least in part, to the pro-inflammatory cytokine production of IL-1ß and TNF-α. The administration of kirenol cream at the dose of 0.3, 0.4 and 0.5% (w/w) significantly inhibited the development of joint swelling induced by CFA, which was auxiliary supported by histopathological studies. CONCLUSION: Kirenol has demonstrated its significant potential to be further investigated for its discovery as a new lead compound for management of topical pain and inflammation, although further pharmacological research is necessary to fully understand its mechanism of action. It also supports the potential beneficial effect of topically administered Siegesbeckia orientalis in inflammatory diseases.

[Flavonoids with special B-ring from Macrothelypteris viridifrons and their anti-proliferative effects on tumor cell].

OBJECTIVE: To study the chemical constituents of Macrothelypteris viridifrons and their anti-proliferative effects on tumor cell. METHOD: The compounds were isolated by column chromatography with silica gel, C18 reverse-phase silica gel, sephadex LH-20, and their structures were elucidated on the basis of physiochemical propertities and spectral analysis. The antitumor activities of all compounds were tested with MOLT4, Hep G2, A-549, MCF-7, HT-29, PC-3 tumor cell lines. RESULT: Five compounds were isolated and identified as protoapigenone (1), protoapigenin (2), protoapigenin-4'-O-beta-D-glucopyanoside (3), 5,7-dihydroxy-2-(1,2-isopropyldioxy-4-oxo-cyclohex-5-enyl) -chromen-4-one (4), 5,7-dihydroxy-2-(1-hydroxy-2,6-dimethoxy-cyclohex-4-oxo) -chromen-4-one (5), respectively. CONCLUSION: All compounds were obtained from this plant for the first time. Compounds 1, 4 and 5 showed strong anti-proliferative effects on six tumor cells, which were in concentration-dependent manner.

In vitro and in vivo evaluation of the wound healing properties of Siegesbeckia pubescens.

ETHNOPHARMACOLOGICAL RELEVANCE: Siegesbeckia pubescens (SP) has been traditionally used as a wound healing agent. AIM OF THE STUDY: Investigate in vitro and in vivo healing properties of SP extract. MATERIALS AND METHODS: The methanolic extract of SP was tested for the ability to stimulate the growth of mouse fibroblast NIH3T3 in vitro. The viability and proliferation of fibroblasts were evaluated at 72 h after cell seeding by MTT assay at 570 nm. To study wound healing properties in vivo, excision and incision wound models were used on rats and SP (3, 4, 5%, w/w) was topically administered. After treatment, wound contraction, epithelialization period and hexosamine content were evaluated in the excision wound model. In the incision wound model, wound sites were removed for histopathological analysis and skin-breaking strength determination. RESULTS: The methanol extract showed significant stimulation of the growth of mouse fibroblast NIH3T3 at 0.5-100 µg/mL. In excision wound, animals treated with 4, 5% (w/w) SP exhibited significant increases in the rate of wound contraction, period of epithelialization and content of hydroxyproline. In incision wound, the animals treated with both the 4 and 5% (w/w) SP extracts showed an increase in breaking strength when compared with the control, which was additionally supported by histopathological studies. CONCLUSION: The experimental data revealed that the methanolic extract of SP displayed remarkable wound healing activity, corroborating its traditional use.

Flavonoids from the aerial parts of Macrothelypteris torresiana.

Two new flavone derivatives (1 and 2) were isolated from the aerial parts of Macrothelypteris torresiana, along with four known flavonoids: protoapigenin, apigenin, kaempferol and quercetin. The structures were determined on the basis of spectroscopic data. Compound 1 showed weak cytotoxic activity against human tumour cell lines HepG2 , MCF7 and K562.

Parathelypteriside attenuates cognition deficits in d-galactose treated mice by increasing antioxidant capacity and improving long-term potentiation.

Parathelypteriside (PG), a stilbenoid compound, was extracted from Parathelypteris glanduligera (kze.) ching that exhibits antioxidative and anti-inflammatory effects. The aim of this study was to investigate the protective effect of PG against the d-galactose (d-gal)-induced neurotoxicity in mice. It was found that long-term intraperitoneal (i.p.) injection of PG (5 or 10 mg/(kg day)) for two weeks significantly improved the behavioral performance of d-gal-treated mice in both Morris water maze test and step-down avoidance test. Biochemical examination revealed that PG reduced the increased levels of malondialdehyde (MDA), and attenuated the decreased activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase in the hippocampus of d-gal-treated mice. Furthermore, the electrophysiological assay showed that PG significantly rescued the long-term potentiation (LTP) impairment in mice hippocampus, and western blotting analysis indicated that the effects of PG on LTP might be attributed to the activation of cAMP-response element-binding protein (CREB). Together, these results suggested that the natural product PG represented a potential source of medicine for the treatment of the neurodegenerative diseases.

Antioxidant, free radical scavenging, anti-inflammatory and hepatoprotective potential of the extract from Parathelypteris nipponica (Franch. et Sav.) Ching.

AIM OF THE STUDY: The present study was conducted to evaluate the antioxidant, free radical scavenging, hepatoprotective and anti-inflammatory potential of Parathelypteris nipponica (Franch. et Sav.) Ching. METHODS AND RESULTS: Antioxidant activity of the methanolic extract of Parathelypteris nipponica (Franch. et Sav.) Ching (TMPN) was studied using in vitro and in vivo models, total flavonoids content in TMPN was found to be 262 +/- 5.6 mg/g (w/w). The TMPN exhibited strong antioxidant activity with EC50 values in reductive ability (0.18 +/- 0.02 mg/ml) and ferric thiocyanate (FTC) assay (0.10 +/- 0.01 mg/ml), strong free radical scavenging activity as evidenced by the low EC50 values in DPPH (1,1-diphenyl-2-picrylhydrazyl) (2.00 +/- 0.02 mg/ml), superoxide anion (0.60 +/- 0.05 mg/ml), OH radicals (0.26 +/- 0.03 mg/ml), and hydrogen peroxide (0.45 +/- 0.03 mg/ml) methods. Acute toxicity study revealed that the LD50 value of the extract was more than the dose 2000 mg/kg bodyweight of mice. Hepatoprotective activity of TMPN was determined by the carbon tetrachloride (CCl4)-induced oxidative tissue injury in rat liver, the extract showed significant hepatoprotective activity that was evident by enzymatic examination and histopathological study. In assessing anti-inflammatory activity the carrageenan-induced rat paw oedema test was used, the extract reduced carrageenan-induced rat paw oedema in dose-dependent manner, achieving high degree of anti-inflammatory activity. CONCLUSION: This study provides a scientific basis for the ethnomedical claims that Parathelypteris nipponica (Franch. et Sav.) Ching is effective against inflammation and liver injury.

Antioxidant and hepatoprotective activity of ethanol extract of Arachniodes exilis (Hance) Ching.

AIM OF THE STUDY: The study was aimed to investigate the ethanol extract of Arachniodes exilis for the antioxidant and hepatoprotective activity. MATERIALS AND METHODS: Antioxidant activity was evaluated by different assays, including reducing power, lipid peroxidation, 2, 2'-diphenyl-1-picrylhydrazyl (DPPH), 2, 2'-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), superoxide anion, hydroxyl radicals and hydrogen peroxide. The hepatoprotective activity of ethanol extract was studied on mice liver damage induced by CCL(4) by monitoring biochemical parameters. RESULTS: The extract showed potent activities on reducing power, lipid peroxide, DPPH, ABTS, superoxide anion, hydroxyl radical and hydrogen peroxide. And oral administration of Arachniodes exilis at different doses resulted in significant improvement on the levels of glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, malondialchehyche and superoxidedismutase. CONCLUSION: The results indicate that this plant possesses potential antioxidant and hepatoprotective properties and has therapeutic potential for the treatment of liver diseases.

Diaqua-(2,6-dihy-droxy-benzoato-κO,O)bis-(2,6-dihy-droxy-benzoato-κO)bis-(1,10-phenanthroline-κN,N')lanthanum(III)-1,10-phenanthroline (1/1).

In the title compound, [La(C(7)H(5)O(4))(3)(C(12)H(8)N(2))(3)(H(2)O)(2)]·C(12)H(8)N(2), the La(III) atom is coordinated by four N atoms from two chelating 1,10-phenanthroline (phen) ligands, four O atoms from three 2,6-dihy-droxy-benzoate (DHB) anions (one monodentate, the other bidentate) and two water O atoms, completing a distorted LaN(4)O(6) bicapped square-anti-prismatic geometry. Within the mononuclear complex mol-ecule, intra-molecular π-π stacking inter-actions are observed, the first between a coordinated phen mol-ecule and a DHB ligand [centroid-centroid distance = 3.7291 (16) Å], and the second between a coordinated phen mol-ecule and an uncoordinated phen ligand [centroid-centroid distance = 3.933 (2) Å]. Inter-molecular π-π stacking is observed between adjacent complexes [inter-planar distance = 3.461 (3) Å]. Intra- and inter-molecular O-H⋯O hydrogen bonds are observed in the DHB ligands and between a water mol-ecule and DHB ligands, respectively. O-H⋯N hydrogen bonds are also observed in the DHB ligands and between uncoordinated phen mol-ecules and aqua ligands.

Bis(2,6-dihy-droxy-benzoato-κO,O)(nitrato-κO,O')bis-(1,10-phenanthroline-κN,N')dysprosium(III).

In the mononuclear title complex, [Dy(C(7)H(5)O(4))(2)(NO(3))(C(12)H(8)N(2))(2)], the Dy(III) atom is in a distorted bicapped square-anti-prismatic geometry formed by four N atoms from two chelating 1,10-phenanthroline (phen) ligands, four O atoms from two 2,6-dihy-droxy-benzoate (DHB) ligands and two O atoms from a nitrate anion. Inter-molecular π-π stacking inter-actions between the phen and DHB ligands [centroid-centroid distances = 3.542 (4) and 3.879 (4) Å] and between the pyridine and benzene rings of adjacent phen ligands [centroid-centroid distance = 3.751 (4) Å] stabilize the crystal structure. Intra-molecular O-H⋯O hydrogen bonds are observed in the DHB ligands.