[Effect of aqueous extract of Corni Fructus on Aß_(25-35)-induced brain injury and neuroinflammation in mice with Alzheimer's disease].

The purpose of this study was to investigate the effect of aqueous extract of Corni Fructus on ß-amyloid protein 25-35(Aß_(25-35))-induced brain injury and neuroinflammation in Alzheimer's disease(AD) mice to provide an experimental basis for the treatment of AD by aqueous extract of Corni Fructus. Sixty C57BL/6J male mice were randomly divided into a sham group, a model group, a positive control group(huperizine A, 0.2 mg·kg~(-1)), a low-dose aqueous extract of Corni Fructus group(1.3 g·kg~(-1)), a medium-dose aqueous extract of Corni Fructus group(2.6 g·kg~(-1)), and a high-dose aqueous extract of Corni Fructus group(5.2 g·kg~(-1)). The AD model was induced by lateral ventricular injection of Aß_(25-35) in mice except for those in the sham group, and AD model mice were treated with corresponding drugs by gavage for 24 days. The behavioral test was performed one week before animal dissection. Hematoxylin-eosin(HE) staining was performed to observe the morphology of neurons in the hippocampal region. Flow cytometry was used to detect the apoptosis level of primary hippocampal cells in mice. ELISA kits were used to detect the levels of ß-amyloid protein 1-42(Aß_(1-42)) and phosphorylated microtubule-associated protein Tau(p-Tau) in mouse brain tissues. Immunofluorescence and Western blot were used to detect the expression of related proteins in mouse brain tissues. MTT assay was used to detect the effect of compounds in aqueous extract of Corni Fructus on Aß_(25-35)-induced N9 cell injury. Molecular docking was employed to analyze the interactions of caffeic acid, trans-p-hydroxy cinnamic acid, isolariciresinol-9'-O-ß-D-glucopyranoside, esculetin, and(+)-lyoniresinol with ß-amyloid precursor protein(APP), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α). Aqueous extract of Corni Fructus could improve the learning and memory abilities of Aß_(25-35)-induced mice by increasing the duration of the autonomous activity, the rate of autonomous alternation, the preference coefficient, and the discrimination coefficient, and reduce Aß_(25-35)-induced brain injury and neuroinflammation in mice by increasing the expression levels of interleukin-10(IL-10) and B-cell lymphoma-2(Bcl-2) in brain tissues, decreasing the expression levels of Aß_(1-42), p-Tau, IL-6, TNF-α, cysteine aspartate-specific protease 3(caspase-3), cysteine aspartate-specific protease 9(caspase-9), and Bcl-2-associated X protein(Bax), and decreasing the number of activated glial cells in brain tissues. The results of cell experiments showed that esculetin and(+)-lyoniresinol could improve Aß_(25-35)-induced N9 cell injury. Molecular docking results showed that caffeic acid, trans-p-hydroxy cinnamic acid, isolariciresinol-9'-O-ß-D-glucopyranoside, esculetin, and(+)-lyoniresinol had good binding affinity with APP and weak binding affinity with IL-6 and TNF-α. Aqueous extract of Corni Fructus could ameliorate cognitive dysfunction and brain damage in Aß_(25-35)-induced mice by reducing the number of apoptotic cells and activated glial cells in the brain and decreasing the expression level of inflammatory factors. Caffeic acid, trans-p-hydroxy cinnamic acid, isolariciresinol-9'-O-ß-D-glucopyranoside, esculetin, and(+)-lyoniresinol may be the material basis for the anti-AD effect of aqueous extract of Corni Fructus.

Role of BDNF-TrkB signaling in the antidepressant-like actions of loganin, the main active compound of Corni Fructus.

AIMS: Corni Fructus (CF) and some CF-contained prescriptions are commonly used in clinical treatment of depression. This investigation aims to evaluate the main active compound of CF in antidepressant properties and its key target. METHODS: Firstly, this study established a behavioral despair model and used high-performance liquid chromatography method to evaluate the antidepressant-like effects of water extract, 20%, 50%, and 80% ethanol extracts of CF, and its main active compound. Then, this study created chronic unpredictable mild stress (CUMS) model to assess loganin's antidepressant-like properties, and its target was evaluated by quantitative real-time polymerase chain reaction, Western blot, Immunofluorescence, enzyme-linked immunosorbent assay, and tyrosine receptor kinase B (TrkB) inhibitor. RESULTS: Results showed that the different extracts of CF significantly shortened the immobility time in forced swimming and tail suspension tests. Moreover, loganin alleviated CUMS-induced depression-like behavior, promoted neurotrophy and neurogenesis, and inhibited neuroinflammation. Furthermore, K252a blocked the improvement of loganin on depression-like behavior, and eliminated the enhancement of neurotrophy and neurogenesis and the inhibition of neuroinflammation. CONCLUSION: Overall, these results indicated that loganin could be used as a major active compound of CF for the antidepressant-like properties and exerted antidepressant-like actions by regulating brain derived neurotrophic factor (BDNF)-TrkB signaling, and TrkB could be used as key target for itsantidepressant-like actions.

Effect of aqueous extract of Corni Fructus on Aβ_(25-35)-induced brain injury and neuroinflammation in mice with Alzheimer's disease / 中国中药杂志

The purpose of this study was to investigate the effect of aqueous extract of Corni Fructus on β-amyloid protein 25-35(Aβ_(25-35))-induced brain injury and neuroinflammation in Alzheimer's disease(AD) mice to provide an experimental basis for the treatment of AD by aqueous extract of Corni Fructus. Sixty C57BL/6J male mice were randomly divided into a sham group, a model group, a positive control group(huperizine A, 0.2 mg·kg~(-1)), a low-dose aqueous extract of Corni Fructus group(1.3 g·kg~(-1)), a medium-dose aqueous extract of Corni Fructus group(2.6 g·kg~(-1)), and a high-dose aqueous extract of Corni Fructus group(5.2 g·kg~(-1)). The AD model was induced by lateral ventricular injection of Aβ_(25-35) in mice except for those in the sham group, and AD model mice were treated with corresponding drugs by gavage for 24 days. The behavioral test was performed one week before animal dissection. Hematoxylin-eosin(HE) staining was performed to observe the morphology of neurons in the hippocampal region. Flow cytometry was used to detect the apoptosis level of primary hippocampal cells in mice. ELISA kits were used to detect the levels of β-amyloid protein 1-42(Aβ_(1-42)) and phosphorylated microtubule-associated protein Tau(p-Tau) in mouse brain tissues. Immunofluorescence and Western blot were used to detect the expression of related proteins in mouse brain tissues. MTT assay was used to detect the effect of compounds in aqueous extract of Corni Fructus on Aβ_(25-35)-induced N9 cell injury. Molecular docking was employed to analyze the interactions of caffeic acid, trans-p-hydroxy cinnamic acid, isolariciresinol-9'-O-β-D-glucopyranoside, esculetin, and(+)-lyoniresinol with β-amyloid precursor protein(APP), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α). Aqueous extract of Corni Fructus could improve the learning and memory abilities of Aβ_(25-35)-induced mice by increasing the duration of the autonomous activity, the rate of autonomous alternation, the preference coefficient, and the discrimination coefficient, and reduce Aβ_(25-35)-induced brain injury and neuroinflammation in mice by increasing the expression levels of interleukin-10(IL-10) and B-cell lymphoma-2(Bcl-2) in brain tissues, decreasing the expression levels of Aβ_(1-42), p-Tau, IL-6, TNF-α, cysteine aspartate-specific protease 3(caspase-3), cysteine aspartate-specific protease 9(caspase-9), and Bcl-2-associated X protein(Bax), and decreasing the number of activated glial cells in brain tissues. The results of cell experiments showed that esculetin and(+)-lyoniresinol could improve Aβ_(25-35)-induced N9 cell injury. Molecular docking results showed that caffeic acid, trans-p-hydroxy cinnamic acid, isolariciresinol-9'-O-β-D-glucopyranoside, esculetin, and(+)-lyoniresinol had good binding affinity with APP and weak binding affinity with IL-6 and TNF-α. Aqueous extract of Corni Fructus could ameliorate cognitive dysfunction and brain damage in Aβ_(25-35)-induced mice by reducing the number of apoptotic cells and activated glial cells in the brain and decreasing the expression level of inflammatory factors. Caffeic acid, trans-p-hydroxy cinnamic acid, isolariciresinol-9'-O-β-D-glucopyranoside, esculetin, and(+)-lyoniresinol may be the material basis for the anti-AD effect of aqueous extract of Corni Fructus.

Cornus iridoid glycoside alleviates sepsis-induced acute lung injury by regulating NF-κB and Nrf2/HO-1 pathways.

BACKGROUND: Sepsis-induced acute lung injury (ALI) is a syndrome associated with inflammation. Cornus iridoid glycoside (CIG), a bioactive component isolated from Corni Fructus, exhibits anti-inflammatory activities. However, the function and underlying mechanisms of CIG in mice with sepsis-induced ALI remain elusive. METHODS: The sepsis-elicited ALI model of mice was established by the induction of cecal ligation and puncture (CLP). The wet/dry (W/D) ratio of lung tissues was examined, and the pathological alterations were determined by hematoxylin and eosin staining. The messenger RNA (mRNA) expressions and serum levels of Interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α (TNF-α) were measured by reverse transcription-quantitative polymerase chain reaction and enzyme-linked immunosorbent serologic assay, respectively. The concentrations of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were assessed by biochemical kits. In addition, the relative protein levels of p-p65, p65, phosphorylated- nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (p-IκBα), IκBα, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) gene were analyzed by Western blotting analysis. RESULTS: CLP enhanced W/D ratio and aggravated pathological changes and scores in mice, which were obviously alleviated by the two concentrations of CIG treatment. CIG treatment notably decreased the CLP-induced mRNA expressions and serum levels of IL-1ß, IL-6, TNF-α, and MDA, but enhanced the decreased concentrations (caused by CLP) of SOD and GSH-Px. Moreover, CIG treatment significantly decreased the ratios of p65/p-p65 and IκBα/p-IκBα caused by CLP, but aggravated the CLP-induced relative protein levels of Nrf2 and HO-1. CONCLUSIONS: CIG obviously ameliorated the sepsis-induced ALI in mice by suppressing inflammation and oxidative stress, which was closely associated with nuclear factor kappa B (NF-κB) and Nrf2-HO-1 signaling pathways.

Cornus iridoid glycoside alleviates sepsis-induced acute lung injury by regulating NF-kB and Nrf2/HO-1 pathways

Background: Sepsis-induced acute lung injury (ALI) is a syndrome associated with inflamma-tion. Cornus iridoid glycoside (CIG), a bioactive component isolated from Corni Fructus, exhibits anti-inflammatory activities. However, the function and underlying mechanisms of CIG in mice with sepsis-induced ALI remain elusive.Methods: The sepsis-elicited ALI model of mice was established by the induction of cecal ligation and puncture (CLP). The wet/dry (W/D) ratio of lung tissues was examined, and the pathological alterations were determined by hematoxylin and eosin staining. The messenger RNA (mRNA) expressions and serum levels of Interleukin (IL)-1Beta IL-6, and tumor necrosis factor-alfa(TNF-alpha) were measured by reverse transcription-quantitative polymerase chain reaction and enzyme-linked immunosorbent serologic assay, respectively. The concentrations of malondial-dehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were assessed by biochemical kits. In addition, the relative protein levels of p-p65, p65, phosphorylated-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (p-IκBalpha), IκBalpha, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) gene were analyzed by Western blotting analysis.Results: CLP enhanced W/D ratio and aggravated pathological changes and scores in mice, which were obviously alleviated by the two concentrations of CIG treatment. CIG treatment notably decreased the CLP-induced mRNA expressions and serum levels of IL-1Beta, IL-6, TNF-alpha, and MDA, but enhanced the decreased concentrations (caused by CLP) of SOD and GSH-Px. Moreover, CIG treatment significantly decreased the ratios of p65/p-p65 and IκBα/p-IκBalpha caused by CLP, but...(AU)

Aqueous extracts of Corni Fructus protect C2C12 myoblasts from DNA damage and apoptosis caused by oxidative stress.

BACKGROUND: Although the various pharmacological effects of Corni Fructus are highly correlated with its antioxidant activity, the blocking effect against oxidative stress in muscle cells is not clear. The purpose of this study was to investigate the effect of aqueous extracts of Corni Fructus (CFE) against oxidative stress caused by hydrogen peroxide (H2O2) in murine skeletal C2C12 myoblasts. METHODS AND RESULTS: MTT assay for cell viability, DCF-DA staining for reactive oxygen species (ROS) production, Comet assay for DNA damage, annexin V-FITC and PI double staining for apoptosis, JC-1 staining and caspase assay for monitor mitochondrial integrity, and western blotting for related protein levels were conducted in H2O2 oxidative stressed C2C12 cells. Our results showed that CFE pretreatment significantly ameliorated the loss of cell viability and inhibited apoptosis in H2O2-treated C2C12 cells in a concentration-dependent manner. DNA damage induced by H2O2 was also markedly attenuated in the presence of CFE, which was associated with suppression of ROS generation. In addition, H2O2 reduced mitochondrial membrane potential and caused downregulation of Bcl-2 and upregulation of Bax expression, although these were abrogated by CFE pretreatment. Moreover, CFE blocked H2O2-induced cytosolic release of cytochrome c, activation of caspase-9 and caspase-3, and degradation of poly (ADP-ribose) polymerase. CONCLUSION: Taken together, the present results demonstrate that CFE could protect C2C12 cells from H2O2-induced damage by eliminating ROS generation, thereby blocking mitochondria-mediated apoptosis pathway. These results indicate that CFE has therapeutic potential for the prevention and treatment of oxidative stress-mediated myoblast injury.

Can cornelian cherry mask bitter taste of probiotic chocolate? Human TAS2R receptors and a sensory study with comprehensive characterisation of new functional product.

Cornelian cherry (Cornus mas L.) fruits are a valuable source of bioactive compounds that are responsible for the perception of bitter taste of chocolate products. The aim of the study was to validate the inhibitory effect of Cornus mas on the TAS2R3 and TAS2R13 bitter taste receptors and to assess the effect of masking the bitter taste of dark chocolate with the help of the sensory panel. Dark chocolate was prepared with an addition of 5% of freeze-dried cornelian cherry fruits and 108 CFU/g of Bacillus coagulans probiotic strains. Effect on the TAS2R receptors was evaluated in specially transfected HEK293T cells, and the inhibition ratio was measured using the calcium release test. Moreover, the total polyphenol content, antioxidant activity and simulated intestinal in vitro digestion were determined for the samples. The tested chocolate products were rich in chlorogenic, caffeic and sinapic acids. The addition of cornelian cherry positively affected the antioxidant activity. The phytochemicals of Cornus mas decreased the TAS2R13 activity by 132% after a 2-minute interaction and, % at the same time, inhibited the TAS2R3 activity by 11.5. Meanwhile, chocolate with the addition of fruit was less bitter according to the sensory panel.

Physico-chemical characterization of Cornus kousa Burg. fruit: determining optimal maturity for fresh consumption.

BACKGROUND: Japanese dogwood (Cornus kousa Burg.) is a popular ornamental plant which develops edible compound fruit utilized in traditional Asian medicine. Previous compositional studies have focused on a small fraction of secondary metabolites at a single maturity stage. In order to address the question of optimal ripeness of Japanese dogwood fruit for consumption the study provides a comprehensive insight into its primary and secondary metabolic profile. RESULTS: The aim of the study was to investigate biochemical composition and morphological traits of Japanese dogwood (Cornus kousa Burg.) fruit at four maturity stages: GF, green fruit; BF, fruit with a defined red blush; RF, ripe fruit; OF, over-ripe fruit. Fruit was characterized by a nearly spherical shape, decreased water content in later stages of ripeness and highest a* values at RF and OF stages. Total sugars increased significantly from GF to OF stage and total organic acids and vitamin C decreased with maturation. Japanese dogwood fruit was characterized by four major phenolic groups: anthocyanins (three), flavonols (eight) hydroxycinnamic acids (three) and flavonoids (one) as well as by four lipophilic antioxidants: tocopherols (two), xanthophylls (five), carotenes (two) and chlorophylls (two). The progression of fruit ripening caused faster accumulation of individual phenolic compounds and lipophilic antioxidants which resulted in significantly higher total phenolic content at the RF and OF stages. CONCLUSION: Japanese dogwood fruit is a rich alternative source of ascorbic acid, α-tocopherol, flavonols and anthocyanins and should be consumed fresh at fully developed red colour of compound berries when their composition is optimal. © 2020 Society of Chemical Industry.

Protective effects of combination of Stauntonia hexaphylla and Cornus officinalis on testosterone-induced benign prostatic hyperplasia through inhibition of 5α- reductase type 2 and induced cell apoptosis.

Benign prostatic hyperplasia (BPH) is a progressive pathological condition associated with proliferation of prostatic tissues, prostate enlargement, and lower-urinary tract symptoms. However, the mechanism underlying the pathogenesis of BPH is unclear. The aim of this study was to investigate the protective effects of a combination of Stauntonia hexaphylla and Cornus officinalis (SC extract) on a testosterone propionate (TP)-induced BPH model. The effect of SC extract was examined in a TP-induced human prostate adenocarcinoma cell line. Male Sprague-Dawley rats were randomly divided into 5 groups (n = 6) for in vivo experiments. To induce BPH, all rats, except those in the control group, were administered daily with subcutaneous injections of TP (5 mg/kg) and orally treated with appropriate phosphate buffered saline/drugs (finasteride/saw palmetto/SC extract) for 4 consecutive weeks. SC extract significantly downregulated the androgen receptor (AR), prostate specific antigen (PSA), and 5α-reductase type 2 in TP-induced BPH in vitro. In in vivo experiments, SC extract significantly reduced prostate weight, size, serum testosterone, and dihydrotestosterone (DHT) levels. Histologically, SC extract markedly recovered TP-induced abnormalities and reduced prostatic hyperplasia, thereby improving the histo-architecture of TP-induced BPH rats. SC extract also significantly downregulated AR and PSA expression, as assayed using immunoblotting. Immunostaining revealed that SC extract markedly reduced the 5α-reductase type 2 and significantly downregulated the expression of proliferating cell nuclear antigen. In addition, immunoblotting of B-cell lymphoma 2 (Bcl-2) family proteins indicated that SC extract significantly downregulated anti-apoptotic Bcl-2 and markedly upregulated pro-apoptotic B cell lymphoma-associated X (Bax) expression. Furthermore, SC treatment significantly decreased the Bcl-2/Bax ratio, indicating induced prostate cell apoptosis in TP-induced BPH rats. Thus, our findings demonstrated that SC extract protects against BPH by inhibiting 5α-reductase type 2 and inducing prostate cell apoptosis. Therefore, SC extract might be useful in the clinical treatment of BPH.

Multiomics-based characterization of specialized metabolites biosynthesis in Cornus Officinalis.

Cornus officinalis, an important traditional medicinal plant, is used as major constituents of tonics, analgesics, and diuretics. While several studies have focused on its characteristic bioactive compounds, little is known on their biosynthesis. In this study, we performed LC-QTOF-MS-based metabolome and RNA-seq-based transcriptome profiling for seven tissues of C. officinalis. Untargeted metabolome analysis assigned chemical identities to 1,215 metabolites and showed tissue-specific accumulation for specialized metabolites with medicinal properties. De novo transcriptome assembly established for C. officinalis showed 96% of transcriptome completeness. Co-expression analysis identified candidate genes involved in the biosynthesis of iridoids, triterpenoids, and gallotannins, the major group of bioactive metabolites identified in C. officinalis. Integrative omics analysis identified 45 cytochrome P450s genes correlated with iridoids accumulation in C. officinalis. Network-based integration of genes assigned to iridoids biosynthesis pathways with these candidate CYPs further identified seven promising CYPs associated with iridoids' metabolism. This study provides a valuable resource for further investigation of specialized metabolites' biosynthesis in C. officinalis.

Functional characterization of Terminal Flower1 homolog in Cornus canadensis by genetic transformation.

KEY MESSAGE: TFL1homologCorcanTFL1suppresses the initiation of inflorescence development and regulates the inflorescence morphology inCornus canadensis. In flowering plants, there is a wide range of variation of inflorescence morphology. Despite the ecological and evolutionary importance, efforts devoted to the evolutionary study of the genetic basis of inflorescence morphology are far fewer compared to those on flower development. Our previous study on gene expression patterns suggested a CorTFL1-CorAP1 based model for the evolution of determinate umbels, heads, and mini dichasia from elongated inflorescences in Cornus. Here, we tested the function of CorcanTFL1 in regulating inflorescence development in Cornus canadensis through Agrobacterium-mediated transformation. We showed that transgenic plants overexpressing CorcanTFL1 displayed delayed or suppressed inflorescence initiation and development and extended periods of vegetative growth. Transgenic plants within which CorcanTFL1 had been down-regulated displayed earlier emergence of inflorescence and a reduction of bract and inflorescence sizes, conversions of leaves to bracts and axillary leaf buds to small inflorescences at the uppermost node bearing the inflorescence, or phyllotaxy changes of inflorescence branches and leaves from decussate opposite to spirally alternate. These observations support an important role of CorcanTFL1 in determining flowering time and the morphological destinies of leaves and buds at the node bearing the inflorescence. The evidence is in agreement with the predicted function of CorTFL1 from the gene expression model, supporting a key role of CorTFL1 in the evolutionary divergence of inflorescence forms in Cornus.

[Transcriptome analysis reveals genes involved in biosynthesis of secondary metabolism in Cornus officinalis].

In order to explore genetic basis for the biosynthesis of secondary metabolism,the transcriptome of Cornus officinalis was sequenced by the new generation of high-throughput sequencing technology,A total of 96 032 unigenes were assembled with an average length of 590.53 bp. Among them, 35 478 unigenes were annotated in the public databases NR,Swissprot,COG,GO,KOG,Pfam and KEGG. Based on the assignment of KEGG pathway, 84 involved in ridoid biosynthesis and 487 unigenes involved in others secondary metabolites biosynthesis were found. Additionally,53 unigenes and 72 unigenes were predicted to have potential functions of cytochome P450 and UDP- glycosyltransferases based on the annotation result, which may encode responsible for secondary metabolites modification. This study was the first comprehensive transcriptome analysis for C. officinalis, and the candidate genes involved in the biosynthesis of secondary metabolites were obtained. The transcriptome data constitutes a much more abundant genetic resource that can be utilized to benefit further molecular biology studies on C. officinalis.

Variation of loganin content in Cornus officinalis fruits at different extraction conditions and maturation stages.

Efficient preparation of loganin from Cornus officinalis fruits was investigated. First, effect of extraction conditions on loganin yield was measured. The loganin content in C. officinalis extract was greatly affected by ethanol concentration and extraction time whereas extraction temperature exerted relatively little effect. Response surface methodology with Box-Behnken design suggested optimized extraction condition for maximum loganin yield as ethanol concentration, 32.0%; temperature 46.2 °C and extraction time, 46.7 min, which yielded 10.4 µg loganin/mg dried fruit. Next, the effect of maturation stage of C. officinalis fruits on loganin content was investigated. The loganin content in the extract of C. officinalis fruits was decreased as the maturation process. The loganin content in the unripe fruits was 18.0 µg/mg extract whereas reduced to 13.3 µg/mg extract for ripe fruits. Taken together, our present study suggested the importance of extraction condition and maturation stages for efficient preparation of loganin from C. officinalis fruits.

Isotopic Differences between Forage Consumed by a Large Herbivore in Open, Closed, and Coastal Habitats: New Evidence from a Boreal Study System.

Documenting habitat-related patterns in foraging behaviour at the individual level and over large temporal scales remains challenging for large herbivores. Stable isotope analysis could represent a valuable tool to quantify habitat-related foraging behaviour at the scale of individuals and over large temporal scales in forest dwelling large herbivores living in coastal environments, because the carbon (δ13C) or nitrogen (δ15N) isotopic signatures of forage can differ between open and closed habitats or between terrestrial and littoral forage, respectively. Here, we examined if we could detect isotopic differences between the different assemblages of forage taxa consumed by white-tailed deer that can be found in open, closed, supralittoral, and littoral habitats. We showed that δ13C of assemblages of forage taxa were 3.0 ‰ lower in closed than in open habitats, while δ15N were 2.0 ‰ and 7.4 ‰ higher in supralittoral and littoral habitats, respectively, than in terrestrial habitats. Stable isotope analysis may represent an additional technique for ecologists interested in quantifiying the consumption of terrestrial vs. marine autotrophs. Yet, given the relative isotopic proximity and the overlap between forage from open, closed, and supralittoral habitats, the next step would be to determine the potential to estimate their contribution to herbivore diet.

Non-Native Ambrosia Beetles as Opportunistic Exploiters of Living but Weakened Trees.

Exotic Xylosandrus spp. ambrosia beetles established in non-native habitats have been associated with sudden and extensive attacks on a diverse range of living trees, but factors driving their shift from dying/dead hosts to living and healthy ones are not well understood. We sought to characterize the role of host physiological condition on preference and colonization by two invaders, Xylosandrus germanus and Xylosandrus crassiusculus. When given free-choice under field conditions among flooded and non-flooded deciduous tree species of varying intolerance to flooding, beetles attacked flood-intolerant tree species over more tolerant species within 3 days of initiating flood stress. In particular, flood-intolerant flowering dogwood (Cornus florida) sustained more attacks than flood-tolerant species, including silver maple (Acer saccharinum) and swamp white oak (Quercus bicolor). Ethanol, a key host-derived attractant, was detected at higher concentrations 3 days after initiating flooding within stems of flood intolerant species compared to tolerant and non-flooded species. A positive correlation was also detected between ethanol concentrations in stem tissue and cumulative ambrosia beetle attacks. When adult X. germanus and X. crassiusculus were confined with no-choice to stems of flood-stressed and non-flooded C. florida, more ejected sawdust resulting from tunneling activity was associated with the flood-stressed trees. Furthermore, living foundresses, eggs, larvae, and pupae were only detected within galleries created in stems of flood-stressed trees. Despite a capability to attack diverse tree genera, X. germanus and X. crassiusculus efficiently distinguished among varying host qualities and preferentially targeted trees based on their intolerance of flood stress. Non-flooded trees were not preferred or successfully colonized. This study demonstrates the host-selection strategy exhibited by X. germanus and X. crassiusculus in non-native habitats involves detection of stress-induced ethanol emission and early colonization of living but weakened trees.

Evidence of physiological decoupling from grassland ecosystem drivers by an encroaching woody shrub.

Shrub encroachment of grasslands is a transformative ecological process by which native woody species increase in cover and frequency and replace the herbaceous community. Mechanisms of encroachment are typically assessed using temporal data or experimental manipulations, with few large spatial assessments of shrub physiology. In a mesic grassland in North America, we measured inter- and intra-annual variability in leaf δ(13)C in Cornus drummondii across a grassland landscape with varying fire frequency, presence of large grazers and topographic variability. This assessment of changes in individual shrub physiology is the largest spatial and temporal assessment recorded to date. Despite a doubling of annual rainfall (in 2008 versus 2011), leaf δ(13)C was statistically similar among and within years from 2008-11 (range of -28 to -27‰). A topography*grazing interaction was present, with higher leaf δ(13)C in locations that typically have more bare soil and higher sensible heat in the growing season (upland topographic positions and grazed grasslands). Leaf δ(13)C from slopes varied among grazing contrasts, with upland and slope leaf δ(13)C more similar in ungrazed locations, while slopes and lowlands were more similar in grazed locations. In 2011, canopy greenness (normalized difference vegetation index - NDVI) was assessed at the centroid of individual shrubs using high-resolution hyperspectral imagery. Canopy greenness was highest mid-summer, likely reflecting temporal periods when C assimilation rates were highest. Similar to patterns seen in leaf δ(13)C, NDVI was highest in locations that typically experience lowest sensible heat (lowlands and ungrazed). The ability of Cornus drummondii to decouple leaf physiological responses from climate variability and fire frequency is a likely contributor to the increase in cover and frequency of this shrub species in mesic grassland and may be generalizable to other grasslands undergoing woody encroachment.

Identification of metabolites of crude and processed Fructus Corni in rats by microdialysis sampling coupled with electrospray ionization linear quadrupole ion trap mass spectrometry.

A microdialysis (MD) sampling coupled with electrospray ionization linear quadrupole ion trap mass spectrometry (LTQ-MS(n)) method has been developed for rapid and sensitive analysis of rat microdialysate metabolite profile of Fructus Corni, a well-known Traditional Chinese Medicine (TCM). The purified samples were separated by a reversed-phase HPLC with C18 column under a gradient elution. Parent compounds and metabolites of crude and processed Fructus Corni of Jiu Zheng Pin (JZP, JZP is produced after steaming the crude drug pre-steeped in wine) were detected by the on-line MS(n) detector in negative scan model. The identification of the metabolites and their structural elucidation were performed by comparing the changes in molecular mass and defining sites of biotransformation based on the accurate MS(n) spectral information of diagnostic fragment ions. In this work, we used such strategies for the identification of the parent compounds and metabolites of crude and processed Fructus Corni in rats, and seven parent compounds and three new metabolites of Fructus Corni were found in rats for the first time. This study provides important structural information regarding to the metabolism of crude Fructus Corni and its JZP. Furthermore, this work also demonstrated the possibilities of using microdialysis sampling coupled with LC-MS(n) approach for identification of bioactive compounds from TCM in vivo.

Competitive uptake and phytomonitoring of chlorinated contaminant mixtures by Redosier dogwood (Cornus sericea).

Plant uptake is an important process in phytoremediation. The robust uptake of volatile organic compounds (VOCs) by plants offers opportunities to establish quantitative relationships between VOCs in plant tissues and in groundwater for the purpose of phytoscreening or phytomonitoring. Most previous research pertaining to phytoremediation neglected the competitive effects of co-contaminants on the uptake of VOCs by plants, yet recent studies appeared to indicate high competitive effects of co-contamination. This study investigated the competitive uptake of three chlorinated compounds in the presence and absence of other co-contaminants by Redosier dogwood in a greenhouse and examined the implications of this competitive phenomenon for phytomonitoring of contaminant mixtures in groundwater. Concentrations of VOCs in stems decreased along the height in both single and bi-solute systems, in agreement with previous observations in the literature. Examination of the VOCs in single and bi-solute systems showed that concentrations of individual compounds are comparable in single and bi-solute systems, yet the ratios of contaminants along the height in bi-solute systems revealed interesting trends. TCE/PCE ratio increased along height while TCE/1,1,2-TCA ratio was roughly constant. The result indicated that sampling point as well as the physicochemical properties of co-contaminants is highly important in phytomonitoring of contaminant mixtures.

Analysis of the interactions of multicomponents in Cornus officinalis Sieb. et Zucc. with human serum albumin using on-line dialysis coupled with HPLC.

Interactions of three iridoid glycosides extracted from Cornus officinalis Sieb. et Zucc. (CIG) with protein were simultaneously explored by on-line dialysis sampling coupled with high-performance liquid chromatography (DS-HPLC). Three main compounds in CIG were unequivocally identified as loganin, sweroside and cornuside by comparing their t(R), MS data and UV spectra with those of reference compounds. Dialysis recoveries and quantitative characteristics of DS-HPLC for three iridoid glycosides were determined. Recoveries of dialysis sampling ranged from 73.9 to 91.7% with the RSD below 3.0%. Based on the determination of concentrations before and after interaction with human serum albumin (HSA), the binding parameters of loganin, sweroside and cornuside with HSA were obtained and the binding mechanisms were investigated.

Cornel iridoid glycoside improves memory ability and promotes neuronal survival in fimbria-fornix transected rats.

Cornel iridoid glycoside (CIG) is a main component extracted from a traditional Chinese herb Cornus officinalis. Our previous study found that CIG improved neurological function in cerebral ischemic rats. The aim of this study was to investigate the therapeutic benefit of CIG in rats with fimbria-fornix transection (FFT) and explore the underlying molecular mechanisms. CIG (20, 60 and 180 mg/kg) or vehicle was intragastrically administered once daily to rats, starting immediately after the surgery and lasting for 4 weeks. Morris water maze and step-through tests showed that the memory deficits seen in FFT rats were significantly improved by CIG treatment. Immunohistochemical analysis showed that CIG treatment attenuated the loss of neurons in hippocampus. To elucidate the memory-improving mechanism of CIG, the neurotrophic factors, synaptic proteins and Bcl-2 family proteins in hippocampus were measured by Western blot analysis. FFT reduced hippocampal protein levels of nerve growth factor (NGF), tyrosine receptor kinase A (Trk A), brain-derived neurotrophic factor (BDNF), synaptophysin (SYP) and B-cell lymphoma-2 (Bcl-2), but not levels of tyrosine receptor kinase B (Trk B) and growth-associated protein 43 (GAP-43). FFT also elevated cytochorome C (Cyt c) and bcl-2-associated X protein (Bax). Administration of CIG to FFT rats significantly elevated the expression of NGF, TrkA, BDNF, SYP, GAP-43 and Bcl-2, and decreased the expression of Cyt c and Bax. These results indicated that CIG effectively counteracted cognitive impairments caused by fimbria-fornix lesions, and the mechanisms might be related to promoting neuronal survival and providing a beneficial environment for brain repair.