Seasonal variation influences flavonoid biosynthesis path and content, and antioxidant activity of metabolites in Tetrastigma hemsleyanum Diels & Gilg.

Environmental conditions contribute to plant growth and metabolism. This study aimed to determine a suitable environment and climate for large-scale artificial cultivation of an endangered plant, Tetrastigma hemsleyanum, by investigating the seasonal variations influencing the flavonoid biosynthetic selectivity and antioxidant activity of its major metabolites. Under conditions of precipitation (2.0~6.6 mm), temperature (17.5~24.1°C), humidity (67.3~80.2%), and sunshine duration (3.4~5.8 h) from April to May, the total flavonoid content in T. hemsleyanum reached higher levels between 281.3 and 392.8 µg/g. In the second half of April, the production selectivity (PS) of isoorientin (IsoO), orientin (Or), rutin (Rut), isoquercitin (IsoQ), kaempferol-3-O-rutinoside (Km3rut), astragalin (Ast), quercetin (Qu), apigenin (Ap), and kaempferol (Km) were 0.30, 0.06, 0.07, 0.07, 0.00, 0.04, 0.38, 0.05, and 0.03, respectively. Naringenin was dehydrogenated or hydroxylated to initiate two parallel reaction pathways for flavonoid biosynthesis in T. hemsleyanum: path I subsequently generated flavone derivatives including apigenin, luteolin, orientin, and isoorientin, and path II subsequently generated flavonol derivatives including Km, Qu, IsoQ, Rut, Ast, and Km3rut. The reaction selectivity of path II (RPSII) from January 1 to September 30 was considerably higher than that of path I (RPSI), except for March 16-31. In addition, either the content or antioxidant activity of three major metabolites in T. hemsleyanum followed the order of phenolic compounds > polysaccharides > sterols, and exhibited dynamic correlations with environmental factors. Naringenin favored hydroxylation and derived six flavonol compounds from January to September, and favored dehydrogenation and derived three flavone compounds from October to December. In most months of a year, Km preferentially favored hydroxylation rather than glucosylation.

Recycling protein selective adsorption on fluorine-modified surface through fluorine-fluorine interaction.

Low surface energy materials with micro-nano structures have been widely developed to prevent non-specific adhesion of biomolecules. Herein we put forward a new approach based on the antifouling and self-assembly properties of fluorine components, to construct a non-specific protein resistance surface with selective protein adsorption property. Briefly, the antifouling surface (SN-F) was obtained by a simple one-step modification on silicon nanowire arrays (SiNWAs) with fluorine coupling agent 1 H,1 H,2 H,2 H-perfluorodecyltrimethoxysilane (FAS). And protein was fluorinated by conjugation with an amphiphilic fluoro-copolymer, produced from 2-methacrylamido glucopyranose (MAG) and trifluoroethyl methacrylate (TFEMA) via RAFT polymerization. The properties of the materials were characterized by 1H nuclear magnetic resonance (1H NMR), infrared spectroscopy (FTIR), water contact angle, and X-ray photoelectron spectroscopy (XPS) etc., and protein adsorption was investigated by protein content measurement, fluorescence detection, and electrophoresis. It was observed that the adsorption for native proteins on SN-F was at an extremely low level, while the adsorption for the fluoro-copolymer conjugated protein (PFG-BSA) was significantly increased. When the percentage of TFEMA in the fluoro-copolymer was as high as 52.0%, the fluorinated protein adsorbed on SN-F was more than 35 times of native proteins on the surface. Moreover, the platform could resist IgG adhesion in serum after the adsorption of fluorinated protein, and it could be recycled three times after 75% ethanol treatment. In conclusion, SN-F showed non-specific protein resistance through low surface energy and specific protein adsorption by fluorine-fluorine self-assembly. The fluorinated nanostructured platform has a great potential in controlling protein adsorption and release.

The essential roles of OsFtsH2 in developing the chloroplast of rice.

BACKGROUND: Filamentation temperature-sensitive H (FtsH) is an ATP-dependent zinc metalloprotease with ATPase activity, proteolysis activity and molecular chaperone-like activity. For now, a total of nine FtsH proteins have been encoded in rice, but their functions have not revealed in detail. In order to investigate the molecular mechanism of OsFtsH2 here, several osftsh2 knockout mutants were successfully generated by the CRISPR/Cas9 gene editing technology. RESULTS: All the mutants exhibited a phenotype of striking albino leaf and could not survive through the stage of three leaves. OsFtsH2 was located in the chloroplast and preferentially expressed in green tissues. In addition, osftsh2 mutants could not form normal chloroplasts and had lost photosynthetic autotrophic capacity. RNA sequencing analysis indicated that many biological processes such as photosynthesis-related pathways and plant hormone signal transduction were significantly affected in osftsh2 mutants. CONCLUSIONS: Overall, the results suggested OsFtsH2 to be essential for chloroplast development in rice.

Theoretical study of twin-column recycling chromatography with a solvent-gradient for preparative binary separations.

Twin-column recycling chromatography with a solvent gradient (TCRC-SG) was investigated with the equilibrium-dispersive chromatography model. The solvent gradient caused by constant addition of a modifier between the two columns created a band compression effect to counterbalance band broadening, so that the target component band neither broadened nor shrunk. Meanwhile, band compression accelerated the separation but prevented excessive separation. Increasing the volume fraction of weak solvent in the modifier and reducing the modifier flowrate enhanced band compression and improved the separation. The effect of column efficiency (number of theoretical plates: 500-1500) on the separation was not significant. According to the separation behavior, a simple operation scheme is proposed to automatically control column switching without needing to determine the adsorption isotherm and designing operating conditions in advance. In comparison with simulated moving bed, TCRC-SG had a higher feed throughput, but consumed more solvent. The results showed that TCRC-SG is favorable for preparative separation.

Interpretation of the mechanism of 3,3'-dichloro-4,4'-diamino diphenylmethane synthesis over HY zeolites.

Catalytic activities of zeolites HY, Hß and HZSM-5 in the heterogeneous synthesis of 3,3'-dichloro-4,4'-diaminodiphenyl methane (MOCA) from o-chloroaniline and formaldehyde were pre-screened in an autoclave, and HY demonstrated better performance than others. Kinetic behaviors of MOCA synthesis over HY(11) were further investigated in a fixed bed continuous flow reactor, and under the conditions of the catalyst bed volume = 20 mL (8.14 g), n(o-chloroaniline) : n(HCHO) = 4 : 1, LHSV = 3.5 h-1, 0.5 MPa and 443 K, HCHO conversion and MOCA selectivity steadily fluctuated at high levels of 90-92% and 75-77% during 16 h, respectively. Catalysts were characterized by BET, NH3-TPD and XRD, products analyzed by HPLC, and reaction intermediates identified by LC/MS and 1H NMR. The mechanism of MOCA synthesis has been interpreted in detail, which also suggested that deposition of basic intermediates on active sites and accumulation of polymeric by-products in pore channels of the catalyst could cause significant decay of HY(11) activity and selectivity under severe conditions. Supplementary tests on catalyst regeneration confirmed that the acidity and surface area of spent HY(11) could be well recovered after burning off the deposited by-products.

Fine optimization of twin-column recycling chromatography with a solvent gradient for the removal of minor impurities.

In the present study, during removal of minor impurities by twin-column recycling chromatography with a solvent gradient, a compressing band effect was generated to offset band spreading and retain the principal component band within one column. For real-time monitoring, a detector was mounted on-line after the upstream column to monitor when the tail of the principal component was eluted from this column. When the conditions fluctuated, the column were switched to ensure successful separation without the need to determine the adsorption equilibrium in advance. Optimization of the operating conditions revealed that increasing the solvent gradient improved pre-impurity separation but impeded post-impurity removal. Changing the feed volume within a certain range hardly affected separation of the impurities, and increasing the number of cycles enhanced separation of the impurities.

Glutathione-Sensitive Silicon Nanowire Arrays for Gene Transfection.

Ingenious surface modification strategies and special topological morphologies endow the biomaterial interface with excellent ability to regulate the cell fate. In this work, a gene delivery platform based on glutathione-sensitive silicon nanowire arrays (SiNWAs) is developed, exhibiting good transfection efficiency of several cell types. Briefly, the surface of SiNWAs is grafted of PEICBA, a branched cationic polymer cross-linked by disulfide bonds (SN-PEICBA). When the cells adhere to the platform surface, silicon nanowires penetrate into the cells and the high concentration of reduced glutathione (GSH) in cytoplasm breaks the disulfide bonds (S-S) in PEICBA. The plasmid DNA preloaded on the cationic polymers is successfully delivered to the nuclei through the nonlysosomal pathway. Cells harvested from the SN-PEICBA show high retention of viability and the platform surface can be reused though S-S replacement for at least three times. In general, our platform is a creative combination of intracellular responsive strategy and surface morphology, which has great potential for auxiliary use in ex vivo cell-based therapies and various biomedical applications.

Optimizing Nucleophilic Depolymerization of Proanthocyanidins in Grape Seeds to Dimeric Proanthocyanidin B1 or B2.

Depolymerization of polymeric proanthocyanidins (PPCs) in grape seeds into oligomeric proanthocyanidins (OPCs), especially the dimers, has important academic significance and practical value. Reaction conditions including nucleophilic reagent/PPC mass ratio, HCl concentration, reaction time, and temperature were systematically optimized by central composite design to maximize the yield of the dimeric product B2 or B1. The yield of B2 reached 3.35 mg mL-1 under the conditions of (-)-epicatechin/PPC mass ratio 2.8, HCl concentration 0.06 mol, reaction time 16 min and temperature 36 °C, and that of B1 reached 3.64 mg mL-1 under the conditions of (+)-catechin/PPC mass ratio 2.8, HCl concentration 0.07 mol, reaction time 17 min, and temperature 34 °C. Overall, this study has provided theoretical guidance and a practical approach to improvethe reaction process and economic value of proanthocyanidins in grape seed proanthocyanidin extract.

Fast and accurate separation of the paclitaxel from yew extracum by a pseudo simulated moving bed with solvent gradient.

A pseudo simulated moving bed (SMB) with solvent gradient was used to trap and separate paclitaxel from yew extracum. This SMB process consisted of three steps: feeding, purification and recovery. In comparison with methanol/water as an eluent, acetonitrile/water could give a better separation but had a poor dissolubility of the yew extracum, and therefore methanol/water was used in the feeding followed by acetonitrile/water in the purification. In the first two steps, water was deliberately added into zone III to modulate the eluotropic strength of the liquid entering zone III, so as to make paclitaxel separation from impurities be more efficient. Once most of impurities discarded, the columns were in turn eluted to recover the trapped paclitaxel of 98% yield with a purity of 78% from the yew extracum containing 1.5% paclitaxel. Afterward, an additional operation of crystallization improved the purity further to 97.8% with the yield of 95%.

Response of Plant Secondary Metabolites to Environmental Factors.

Plant secondary metabolites (SMs) are not only a useful array of natural products but also an important part of plant defense system against pathogenic attacks and environmental stresses. With remarkable biological activities, plant SMs are increasingly used as medicine ingredients and food additives for therapeutic, aromatic and culinary purposes. Various genetic, ontogenic, morphogenetic and environmental factors can influence the biosynthesis and accumulation of SMs. According to the literature reports, for example, SMs accumulation is strongly dependent on a variety of environmental factors such as light, temperature, soil water, soil fertility and salinity, and for most plants, a change in an individual factor may alter the content of SMs even if other factors remain constant. Here, we review with emphasis how each of single factors to affect the accumulation of plant secondary metabolites, and conduct a comparative analysis of relevant natural products in the stressed and unstressed plants. Expectantly, this documentary review will outline a general picture of environmental factors responsible for fluctuation in plant SMs, provide a practical way to obtain consistent quality and high quantity of bioactive compounds in vegetation, and present some suggestions for future research and development.

Nup62, associated with spindle microtubule rather than spindle matrix, is involved in chromosome alignment and spindle assembly during mitosis.

An increasing number of the active mitotic functions of nucleoporins in the distinct steps of mitosis have been assigned over the past few years. As one of FG-repeats containing nucleoporins, Nup62 has been found to be involved in nuclear transport, cell migration, virus infection, and cell cycle regulation. However, the role and mechanism of Nup62 in mitotic regulation have not been fully revealed. In this paper, it was revealed that a fraction of Nup62 was associated with mitotic spindle microtubule instead of spindle matrix, and the localization of Nup62 in the mitotic spindle depended on its three coiled-coil domains rather than Crm1, although Nup62 strongly interacted with Crm1 during mitosis. Moreover, depletion of Nup62 by small interference of RNA seriously induced the defects of chromosome alignment and spindle assembly although the bipolar spindle was observed in most of the Nup62 knock-down cells. Notably, congression of polar chromosome defect was observed in more than 30% of Nup62 knock-down cells. These findings revealed that Nup62 was a novel mitotic spindle associated nucleoporin and involved in chromosome alignment and spindle assembly.

Study on a pseudo-simulated moving bed with solvent gradient for ternary separations.

There is a trapping effect on the solute in the simulated moving bed (SMB) with solvent gradient. As the eluotropic strength of the liquid in zone II (between the extract-port and feed-port) is higher than that in zone III (between the feed-port and the raffinate-port), the solute can move forward in zone II but backward in zone III to be trapped in the two zones consequently. On this basis, a pseudo-SMB was proposed to separate the medium retained solute from a ternary mixture by selectively trapping the medium retained solute. Once the columns in zones II and III are saturated with the target solute, the solvent dissolving the feed is introduced at the feed-port to remove the least retained solute from the raffinate-port and the most retained solute from the extract-port. Finally, the target solute is recovered. A model simulation was used to investigate the behavior and performance of a system operating with this technology. The results showed that increasing the columns in zone III could trap more target solutes since the eluotropic strength decreased from zone II to zone III. Partial recovery in combination with incomplete purification could improve the separation performance in terms of the productivity and solvent consumption without losing purity.

A novel pseudo simulated moving bed with solvent gradient for ternary separations.

A novel pseudo simulated moving bed was suggested to separate a ternary mixture. A solvent gradient was created to make the solvent strength decreasing from zone II to zone III. Under suitable conditions, the least retained solute A moved forward and the most retained solute C moved backward in zones II and III whereas the medium retained solute B moved forward in zone II but backward in zone III to be trapped in the two zones consequently. Once the columns in zones II and III were saturated with solute B, the solvent dissolving the feed was introduced at the feed port to remove solute A from the raffinate-port and solute C from the extract-port. Finally, solute B was recovered from the extract port by stopping the liquid flow in zone II. This scheme was validated by the successful separation of dihydrocapsaicin from capsaicinoids.

Protective effects and mechanism of puerarin on learning-memory disorder after global cerebral ischemia-reperfusion injury in rats.

OBJECTIVE: To observe the effect of puerarin on the learning-memory disorder after global cerebral ischemia-reperfusion injury in rats, and to explore its mechanism of action. METHODS: The global cerebral ischemia-reperfusion injury model was established using the modifified Pulsinelli four-vessel occlusion in Sprague-Dawley rats. Rats were intraperitoneally injected with puerarin (100 mg/kg) 1 h before ischemia and once every 6 h afterwards. The learning-memory ability was evaluated by the passive avoidance test. The dynamic changes of the cell counts of apoptosis and positive expression of Bcl-2 in the hippocampus CA1 region were determined by the TUNEL and immunohistochemical methods, respectively. RESULTS: (1) Compared with the reperfusion group, the step through latency (STL) in the passive avoidance test in the puerarin group was prolonged signifificantly (P<0.01). (2) The apoptotic neurons were injured most severely on the 3rd day in the hippocampal CA1 region after global ischemia and reperfusion. In the puerarin group, the number of apoptotic cells decreased at respective time points after ischemia-reperfusion (P<0.01). (3) The level of positive expression of Bcl-2 varied according to the duration of reperfusion and the peak level occurred on day 1 in the hippocampal CA1 region after global cerebral ischemia. Compared with the reperfusion group, the expression of Bcl-2 in the puerarin group was up-regulated at the respective time points after ischemia reperfusion (P<0.01), reaching the peak on day 1. CONCLUSIONS: Puerarin could improve the learning-memory ability after global cerebral ischemia and reperfusion in rats. The protective mechanism might be related to the effect of inhibiting or delaying the cell apoptosis through up-regulating the expression of Bcl-2 after ischemia and reperfusion.

Separation of capsaicin from capsaicinoids by simulated moving bed chromatography.

Capsaicinoids were separated from capsicum oleoresin by solvent extraction and adsorption separation on macropore resin, and used directly as the feed of simulated moving bed. Using a mobile phase of methanol/water (75/25, v/v) and ODS columns, the two key components, capsaicin and dihydrocapsaicin, were separated completely, while part of minor weak impurities were discarded by forcing them to leak into zone I. The amount of discarded impurities increases with decreasing flow rate in zone I so that the purity of capsaicin in raffinate stream could be improved.