Isolation and Identification of the High-Affinity DNA Aptamer Target to the Brain-Derived Neurotrophic Factor (BDNF).

Aptamers are functional oligonucleotide ligands used for the molecular recognition of various targets. The natural characteristics of aptamers make them an excellent alternative to antibodies in diagnostics, therapeutics, and biosensing. DNA aptamers are mainly single-stranded oligonucleotides (ssDNA) that possess a definite binding to targets. However, the application of aptamers to the fields of brain health and neurodegenerative diseases has been limited to date. Herein, a DNA aptamer against the brain-derived neurotrophic factor (BDNF) protein was obtained by in vitro selection. BDNF is a potential biomarker of brain health and neurodegenerative diseases and has functions in the synaptic plasticity and survival of neurons. We identified eight aptamers that have binding affinity for BDNF from a 50-nucleotide library. Among these aptamers, NV_B12 showed the highest sensitivity and selectivity for detecting BDNF. In an aptamer-linked immobilized sorbent assay (ALISA), the NV_B12 aptamer strongly bound to BDNF protein, in a dose-dependent manner. The dissociation constant (Kd) for NV_B12 was 0.5 nM (95% CI: 0.4-0.6 nM). These findings suggest that BDNF-specific aptamers could be used as an alternative to antibodies in diagnostic and detection assays for BDNF.

Standardized microwave extract of Sappan Lignum exerts anti­inflammatory effects through inhibition of NF­κB activation via regulation of heme oxygenase­1 expression.

The extract of Sappan Lignum, the heartwood of Caesalpinia sappan L., has been used in medicine to improve blood circulation. Recently, the application of microwave extraction methods has been a major focus of research into the extraction of components from natural sources. In this experiment, we compared the anti­inflammatory effects of Sappan Lignum prepared by heat­70% EtOH extraction (CSE­H­70E) and microwave­70% EtOH extraction (CSE­MW­70E). High­performance liquid chromatography analysis was used to identify the compounds in these extracts. The heat­70% EtOH and microwave­70% EtOH extracts of Sappan Lignum had different chromatograms. CSE­MW­70E significantly inhibited the protein expression of iNOS and COX­2, PGE2, TNF­α, and reduced NO and IL­1ß production in macrophages exposed to LPS, whereas, only high concentrations of CSE­H­70E (20 µg/ml) resulted in any effects. Furthermore, CSE­MW­70E upregulated heme oxygenase­1 (HO­1) expression. In addition, the use of tin protoporphyrin, an inhibitor of HO­1, confirmed the inhibitory effects of CSE­MW­70E on pro­inflammatory mediators. These results suggested that the CSE­MW­70E­mediated upregulation of HO­1 played an important role in the anti­inflammatory effects of macrophages. Therefore, these findings showed that microwave extraction can be utilized to improve the extraction efficiency and biological activity of Sappan Lignum.

Oleanane triterpenoids from Akebiae Caulis exhibit inhibitory effects on Aß42 induced fibrillogenesis.

Previous phytochemical investigations of Akebiae Caulis resulted in the isolation of triterpenes, triterpene glycosides, phenylethanoid glycosides and megastigmane glycoside. Amyloid beta (Aß), the main component of the senile plaques detected in Alzheimer's disease, induces cell death. However, only a limited number of studies have addressed the biological and pharmacological effects of Akebiae Caulis. In particular, the inhibitory activity of Akebiae Caulis against Aß42 fibrillogenesis remains unclear. Herein, a new triterpene glycoside, akequintoside F (1), along with nine known compounds pulsatilla saponin A (2), collinsonidin (3), akebonic acid (4), hederagenin (5), 1-(3',4'-dihydroxycinnamoyl) cyclopentane-2,3-diol (6), asperosaponin C (7), leontoside A (8), quinatic acid (9), and quinatoside A (10) were isolated from Akebiae Caulis using repeated column chromatography with silica gel, LiChroprep RP-18, and MCI gel. The chemical structures of compounds 1-10 were illustrated based on 1D and 2D NMR spectroscopy, including 1H-1H COSY, HSQC, HMBC and NOESY spectroscopic analyses. Compound 1 a novel compound and known compounds 6 and 7 were isolated for the first time from this plant. Among these compounds, 1, 3, 4, 5 and 7 displayed significant inhibitory effects on Aß42 induced fibrillogenesis. We present the first report of new compound 1 and the inhibitory effects of components from Akebiae Caulis on Aß42 fibrillogenesis.

A new indole glycoside from the seeds of Raphanus sativus.

A new indole glycoside, ß-D-glucopyranosyl 2-(methylthio)-1H-indole-3-carboxylate, named raphanuside A (1), as well as eight known compounds, ß-D-fructofuranosyl-(2 â†’ 1)-(6-O-sinapoyl)-α-D-glucopyranoside (2), (3-O-sinapoyl)-ß-D-fructofuranosyl-(2 â†’ 1)-α-D-glucopyranoside (3), (3-O-sinapoyl)-ß-D-fructofuranosyl-(2 â†’ 1)-(6-O-sinapoyl)-α-D-glucopyranoside (4), (3,4-O-disinapoyl)-ß-D-fructofuranosyl-(2 â†’ 1)-(6-O-sinapoyl)-α-D-glucopyranoside (5), isorhamnetin 3,4'-di-O-ß-D-glucoside (6), isorhamnetin 3-O-ß-D-glucoside-7-O-α-L-rhamnoside (7), isorhamnetin 3-O-ß-D-glucoside (8) and 3'-O-methyl-(-)-epicatechin 7-O-ß-D-glucoside (9) were isolated from the seeds of Raphanus sativus. Furthermore, compounds 1-3 and 6-9, were isolated from this plant for the first time. The structures of compounds 1-9 were identified using 1D and 2D NMR, including (1)H-(1)H COSY, HSQC, HMBC and NOESY spectroscopic analyses. The inhibitory activity of these isolated compounds against interleukin-6 (IL-6) production in TNF-α stimulated MG-63 cells was also examined.

Evaluation of Antidiarrheal Activity of Methanolic Extract of Maranta arundinacea Linn. Leaves.

Diarrhea is one of the most common causes for thousands of deaths every year. Therefore, identification of new source of antidiarrheal drugs becomes one of the most prominent focuses in modern research. Our aim was to investigate the antidiarrheal and cytotoxic activities of methanolic extract of Maranta arundinacea linn. (MEMA) leaves in rats and brine shrimp, respectively. Antidiarrheal effect was evaluated by using castor oil-induced diarrhea, enteropooling, and gastrointestinal motility tests at 200 mg/kg and 400 mg/kg body weight in rats where the cytotoxic activity was justified using brine shrimp lethality bioassay at different concentrations of MEMA. The extract showed considerable antidiarrheal effect by inhibiting 42.67% and 57.75% of diarrheal episode at the doses of 200 and 400 mg/kg, respectively. MEMA also significantly (p < 0.01) reduced the castor oil-induced intestinal volume (2.14 ± 0.16 to 1.61 ± 0.12 mL) in enteropooling test as well as intestinal transit (33.00 to 43.36%) in GI motility test, compared to their respective control. These observed effects are comparable to that of standard drug loperamide (5 mg/kg). On the other hand, in brine shrimp lethality test after 24 h, surviving brine shrimp larvae were counted and LD50 was assessed. Result showed that MEMA was potent against brine shrimp with LD50 value of 420 µg/mL. So the highest dose of 400 µg/mL of MEMA was not toxic to mice. So these results indicate that bioactive compounds are present in methanolic extract of Maranta arundinacea leaves including significant antidiarrheal activity and could be accounted for pharmacological effects.

Chemical constituents on the aerial parts of Artemisia selengensis and their IL-6 inhibitory activity.

Ten compounds, 1',3'-propanediol,2'-amino-1'-(1,3-benzodioxol-5-yl) (1), artanomaloide (2), canin (3), eupatilin (4), quercetin-3-O-ß-D-glucoside-7-O-α-L-rhamnoside (5), 1,3-di-O-caffeoylquinic acid (6), isoquercitrin (7), pinoresinol-4-O-ß-D-glucoside (8), scopolin (9), and isofraxidin-7-O-ß-D-glucopyranoside (10) were isolated from the aerial parts of A. selengensis. The structures of compounds (1-10) were identified based on 1D and 2D NMR, including (1)H-(1)H COSY, HSQC, HMBC and NOESY spectroscopic analyses. Among them, compound 1 was isolated from this plant for the first time as a naturally occurring compound. The inhibitory activity of these isolated compounds against interleukin-6 (IL-6) production in TNF-α stimulated MG-63 cells was also examined.