Crateva unilocularis Buch. shoots attenuate D-galactose-induced brain injury and cognitive disorders of mice through the PI3K/Akt/Nrf2 pathway.

Crateva unilocularis and C. unilocularis Buch. shoots are traditionally served as a vegetable with many health-promoting benefits. The aim of the current investigation was designed to study the preventive effect of ethanol extract from C. unilocularis shoots (CSE) on D-galactose (D-gal) induced brain and liver injury and cognitive disorders of mice, and to illuminate the potential molecular mechanisms. Results exhibited that CSE, especially at a dose of 600 mg kg-1 b.w., remarkably improved the morphology and number of hippocampal neurons against impairment caused by D-gal (p < 0.05). Biochemical parameters displayed that CSE, especially at a high dose, substantially increased SOD (by about 35.37%, 24.98% and 45.51%), CAT (by about 45.73%, 31.44% and 30.70%), and GSH-Px (by about 36.47%, 17.68% and 52.87%) activities and decreased the level of MDA (by about 22.58%, 23.38% and 27.69%) in the plasma, liver, and brain, respectively (p < 0.05). The AChE content in mice administered a high dose of CSE decreased by about 44.48%. Further mechanism results revealed that the prevention by CSE might be achieved by upregulating the expressions of p-PI3K/PI3K, p-Akt/Akt, Nrf2, HO-1, and NQO1 proteins (p < 0.05). In addition, behavioral experiments showed that CSE at a dose of 600 mg kg-1 could significantly improve the anxiety status and impairment of learning and memory caused by D-gal (p < 0.05). These results substantially proved that C. unilocularis shoots could effectively attenuate D-gal-induced brain and liver injury and cognitive disorders of mice and could be applied to develop new functional foods to encounter aging-related impairments in brain function.

A Rare Dirhamnosyl Flavonoid and Other Radical-Scavenging Metabolites from Cynophalla mollis (Kunth) J.†Presl and Colicodendron scabridum (Kunt) Seem. (Capparaceae) of Ecuador.

The phytochemistry of Cynophalla mollis (Kunth) J. Presl and Colicodendron scabridum (Kunth), both belonging to the family Capparaceae, were investigated in this study for the first time. Lupeol, betulin, lutein, stachydrine and quercetin-3,4'-di-O-rhamnoside were isolated from C. mollis, whereas C. scabridum afforded lupeol, lutein, stachydrine, ß-sitosterol, stigmasterol, betonicine and narcissoside. All these compounds were purified by preparative liquid chromatography, in both open column and instrumental (MPLC) separation systems. Preparative TLC was also applied. They were all identified by 1 H- and 13 C-NMR experiments. The complete structure of the very rare flavonoid quercetin-3,4'-di-O-rhamnoside was fully elucidated through DEPT-135, COSY, HMQC and HMBC experiments, together with UV/VIS and FT-IR spectrophotometry. Complete NMR data for quercetin-3,4'-di-O-rhamnoside in deuterated methanol were presented here for the first time. All the extracts did not exert antioxidant activity at the maximum tested dose of 1 mg/mL. Three out of the nine isolated compounds exerted a good spectrum of antioxidant capacity, being narcissoside the most active against ABTS radicals, with SC50 =12.43 µM. It was followed by lutein and quercetin-3,4'-di-O-rhamnoside, with 40.92 µM and 46.10 µM, respectively.

Crataeva tapia bark lectin (CrataBL) is a chemoattractant for endothelial cells that targets heparan sulfate and promotes in vitro angiogenesis.

Formation of new blood vessels from preexisting ones, a process known as angiogenesis, is one of the limiting steps for success in treatment of ischemic disorders. Therefore, efforts to understanding and characterize new agents capable to stimulate neovascularization are a worldwide need. Crataeva tapia bark lectin (CrataBL) has been shown to have chemoattractant properties for endothelial cells through the stimulation of migration and invasiveness of human umbilical vein endothelial cells (HUVEC) because it is a positively charged protein with high affinity to glycosaminoglycan. In addition, CrataBL increased the production of chondroitin and heparan sulfate in endothelial cells. These findings orchestrated specific adhesion on collagen I and phosphorylation of tyrosine kinase receptors, represented by vascular endothelial growth factor receptor-2 (VEGFR-2) and fibroblast growth factor receptor (FGFR), whose downstream pathways trigger the angiogenic cascade increasing cell viability, cytoskeleton rearrangement, cell motility, and tube formation. Moreover, CrataBL inhibited the activity of matrix metalloproteases type 2 (MMP-2), a protein related to tissue remodeling. Likewise, CrataBL improved wound healing and increased the number of follicular structures in lesioned areas produced in the dorsum-cervical region of C57BL/6 mice. These outcomes altogether indicate that CrataBL is a pro-angiogenic and healing agent.

Arbuscular mycorrhizal fungi improve photosynthetic energy use efficiency and decrease foliar construction cost under recurrent water deficit in woody evergreen species.

Plants suffer recurrent cycles of water deficit in semiarid regions and have several mechanisms to tolerate low water availability. Thus, arbuscular mycorrhizal fungi (AMF) can alleviate deleterious effects of stress. In this study, Cynophalla flexuosa plants, a woody evergreen species from semiarid, when associated with AMF were exposed to two consecutive cycles of water deficit. Leaf primary metabolism, specific leaf area (SLA), leaf construction cost (CC) and photosynthetic energy use efficiency (PEUE) were measured. The maximum stress occurred on seven days (cycle 1) and ten days (cycle 2) after suspending irrigation (photosynthesis close to zero). The rehydration was performed for three days after each maximum stress. In both cycles, plants submitted to water deficit showed reduced gas exchange and leaf relative water content. However, Drought + AMF plants had significantly larger leaf relative water content in cycle 2. At cycle 1, the SLA was larger in non-inoculated plants, while CC was higher in inoculated plants. At cycle 2, Drought + AMF treatment had lower CC and large SLA compared to control, and high PEUE compared to Drought plants. These responses suggest AMFs increase tolerance of C. flexuosa to recurrent water deficit, mainly in cycle 2, reducing the CC, promoting the improvement of SLA and PEUE, leading to higher photosynthetic area. Thus, our result emphasizes the importance of studies on recurrence of water deficit, a common condition in semiarid environments.

Protective action of Crateva nurvala Buch. Ham extracts against renal ischaemia reperfusion injury in rats via antioxidant and anti-inflammatory activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Crateva nurvala stem bark is commonly used in Ayruveda in treatment of many renal injuries, e.g., urinary lithiasis, diuretic and nephroprotective. However, its protective effect against renal ischaemia/reperfusion, the major cause of acute kidney injury, has never been studied. Moreover, no comprehensive chemical profiling of its extracts was recorded. AIM OF THE STUDY: Assessment of the protective effect of the plant extracts against renal ischaemia/reperfusion and elucidation of the possible mechanism of action. Then, to determine its bioactive constituents using modern UPLC-HRMS technique. MATERIAL AND METHODS: Unilateral ischaemia was induced by clamping the left renal artery for 1h then reperfusion for 24h. Rats were divided in 4 groups: i) sham-operated group, ii) ischaemia/reperfusion, I/R group, iii) I/R protected by previous administration of Crateva leaves extract, CLE group and iv) I/R protected by previous administration of Crateva bark extract, CBE group. At the end of reperfusion, blood samples were analyzed for renal function biomarkers. Kidneys were examined histopathologically and their homogenates were used in determining the intracellular levels of oxidative stress, inflammatory, and apoptosis markers. RESULTS: Leaves and bark extracts attenuated the deleterious effects of I/R apparent in reducing LDH, creatinine and blood urea nitrogen levels. The extracts reduced the oxidative stress by replenishing the glutathione levels and Nrf2 factor levels. Moreover, extracts decreased levels of pro-inflammatory TNF-α, NF-κß and IL-6; which ultimately resulted in reducing the pro-apoptotic caspase-3. Bark and leave extracts have quite similar chemical profile where 42 compounds of various chemical classes were identified. Flavonoids are the major class of the bioactive phytochemicals CONCLUSION: C. nurvala extracts had effectively ameliorated the deleterious effects of renal I/R by mainly counteracting oxidative stress and presumably inflammation. Consequently, it can be used as a complementary treatment with other agents. In this aspect, leaves stand as a sustainable alternative to bark. The presented chemical profiling can be used in future standardization and quality control of the drug.

Differential distribution of amino acids in plants.

Plants are a rich source of amino acids and their individual abundance in plants is of great significance especially in terms of food. Therefore, it is of utmost necessity to create a database of the relative amino acid contents in plants as reported in literature. Since in most of the cases complete analysis of profiles of amino acids in plants was not reported, the units used and the methods applied and the plant parts used were different, amino acid contents were converted into relative units with respect to lysine for statistical analysis. The most abundant amino acids in plants are glutamic acid and aspartic acid. Pearson's correlation analysis among different amino acids showed that there were no negative correlations between the amino acids. Cluster analysis (CA) applied to relative amino acid contents of different families. Alismataceae, Cyperaceae, Capparaceae and Cactaceae families had close proximity with each other on the basis of their relative amino acid contents. First three components of principal component analysis (PCA) explained 79.5% of the total variance. Factor analysis (FA) explained four main underlying factors for amino acid analysis. Factor-1 accounted for 29.4% of the total variance and had maximum loadings on glycine, isoleucine, leucine, threonine and valine. Factor-2 explained 25.8% of the total variance and had maximum loadings on alanine, aspartic acid, serine and tyrosine. 14.2% of the total variance was explained by factor-3 and had maximum loadings on arginine and histidine. Factor-4 accounted 8.3% of the total variance and had maximum loading on the proline amino acid. The relative content of different amino acids presented in this paper is alanine (1.4), arginine (1.8), asparagine (0.7), aspartic acid (2.4), cysteine (0.5), glutamic acid (2.8), glutamine (0.6), glycine (1.0), histidine (0.5), isoleucine (0.9), leucine (1.7), lysine (1.0), methionine (0.4), phenylalanine (0.9), proline (1.1), serine (1.0), threonine (1.0), tryptophan (0.3), tyrosine (0.7) and valine (1.2).

Post-transcriptional regulation of photosynthetic genes is a key driver of C4 leaf ontogeny.

C4 photosynthesis allows highly efficient carbon fixation that originates from tightly regulated anatomical and biochemical modifications of leaf architecture. Recent studies showed that leaf transcriptome modifications during leaf ontogeny of closely related C3 (Tarenaya hassleriana) and C4 (Gynandropsis gynandra) species within the Cleomaceae family existed but they did not identify any dedicated transcriptional networks or factors specifically driving C4 leaf ontogeny. RNAseq analysis provides a steady-state quantification of whole-cell mRNAs but does not allow any discrimination between transcriptional and post-transcriptional processes that may occur simultaneously during leaf ontogeny. Here we use exon-intron split analysis (EISA) to determine the extent to which transcriptional and post-transcriptional processes are involved in the regulation of gene expression between young and expanded leaves in both species. C4-specific changes in post-transcriptional regulation were observed for genes involved in the Calvin-Benson cycle and some photosystem components but not for C4 core-cycle genes. Overall, this study provides an unbiased genome-wide insight into the post-transcriptional mechanisms that regulate gene expression through the control of mRNA levels and could be central to the onset of C4 photosynthesis. This mechanism is cytosolic which implies cell-specific modifications of mRNA stability. Understanding this mechanism may be crucial when aiming to transform C3 crops into C4 crops.

Structural studies of the interaction of Crataeva tapia bark protein with heparin and other glycosaminoglycans.

CrataBL, a protein isolated from Crataeva tapia bark, which is both a serine protease inhibitor and a lectin, has been previously shown to exhibit a number of interesting biological properties, including anti-inflammatory, analgesic, antitumor, and insecticidal activities. Using a glycan array, we have now shown that only sulfated carbohydrates are effectively bound by CrataBL. Because this protein was recently shown to delay clot formation by impairing the intrinsic pathway of the coagulation cascade, we considered that its natural ligand might be heparin. Heparin is a glycosaminoglycan (GAG) that interacts with a number of proteins, including thrombin and antithrombin III, which have a critical, essential pharmacological role in regulating blood coagulation. We have thus employed surface plasmon resonance to improve our understanding of the binding interaction between the heparin polysaccharide and CrataBL. Kinetic analysis shows that CrataBL displays strong heparin binding affinity (KD = 49 nM). Competition studies using different size heparin-derived oligosaccharides showed that the binding of CrataBL to heparin is chain length-dependent. Full chain heparin with 40 saccharides or large oligosaccharides, having 16-18 saccharide residues, show strong binding affinity for CrataBL. Heparin-derived disaccharides through tetradecasaccharides show considerably lower binding affinity. Other highly sulfated GAGs, including chondroitin sulfate E and dermatan 4,6-disulfate, showed CrataBL binding affinity comparable to that of heparin. Less highly sulfated GAGs, heparan sulfate, chondroitin sulfate A and C, and dermatan sulfate displayed modest binding affinity as did chondroitin sulfate D. Studies using chemically modified heparin show that N-sulfo and 6-O-sulfo groups on heparin are essential for CrataBL-heparin interaction.

Cellular and subcellular compartmentation of Ni in the Eurasian serpentine plants Alyssum bracteatum, Alyssum murale (Brassicaceae) and Cleome heratensis (Capparaceae).

This study investigated the cellular and subcellular compartmentation of Ni in the Eurasian serpentine species Alyssum murale, Alyssum bracteatum and Cleome heratensis and a non-serpentine population of A. murale (as a control) grown in hydroponic culture. Plant growth responses and Ni uptake clearly revealed the higher Ni tolerance of serpentine plants than the non-serpentine plants. Serpentine A. murale and A. bracteatum grew better at elevated (0.01 mM) Ni in the nutrient solution, supporting the view that the Ni hyperaccumulators have a higher requirement for Ni than normal plants. Low shoot Ni content of C. heratensis in response to the high Ni treatments indicated that this species employs an avoidance strategy for Ni tolerance. Energy-dispersive X-ray microanalysis showed that Ni was highly concentrated in the cell walls and cell lumen, most likely the vacuoles, of leaf epidermis of A. murale and A. bracteatum rather than in the mesophyll cells. EDX spectra from leaves of the non-serpentine A. murale suggested that Ni accumulated in both epidermal and mesophyll cells but not in the epidermal cell walls. Growth reduction and Ni toxicity in plants of the non-serpentine A. murale could be due to accumulation of Ni in the lumen of leaf mesophyll cells. Our data suggest that cellular and subcellular compartmentation are both possible mechanisms for Ni tolerance employed by the serpentine A. murale and A. bracteatum.