Synthesis, biological activity and mechanism of action of novel allosecurinine derivatives as potential antitumor agents.

Cancer with low survival rates is the second main cause of death among all diseases in the world and consequently, effective antineoplastic agents are urgently needed. Allosecurinine is a plant-derived indolicidine securinega alkaloid shown bioactivity. The object of this study is to investigate synthetic allosecurinine derivatives with considerable anticancer capacity against nine human cancer cell lines as well as mechanism of action. We synthesized twenty-three novel allosecurinine derivatives and evaluated their antitumor activity against nine cancer cell lines for 72 h by MTT and CCK8 assays. FCM was applied to analyze the apoptosis, mitochondrial membrane potential, DNA content, ROS production, CD11b expression. Western blot was selected to analyze the protein expression. Structure-activity relationships were established and potential anticancer lead BA-3 which induced differentiation of leukemia cells towards granulocytosis at low concentration and apoptosis at high concentration was identified. Mechanism studies showed that mitochondrial pathway mediated apoptosis within cancer cells with cell cycle blocking was induced by BA-3. In addition, western blot assays revealed that BA-3 induced expression of the proapoptotic factor Bax, p21 and reduced the levels of antiapoptotic protein such as Bcl-2, XIAP, YAP1, PARP, STAT3, p-STAT3, and c-Myc. Collectively, BA-3 was a lead compound for oncotherapy at least in part, through the STAT3 pathway. These results were an important step in further studies on allosecurinine-based antitumor agent development.

Effects of P-Coumarate 3-Hydroxylase Downregulation on the Compositional and Structural Characteristics of Lignin and Hemicelluloses in Poplar Wood (Populus alba × Populus glandulosa).

Elucidating the chemical and structural characteristics of hemicelluloses and lignin in the p-coumarate 3-hydroxylase (C3H) down-regulated poplar wood will be beneficial to the upstream gene validation and downstream biomass conversion of this kind of transgenic poplar. Herein, the representative hemicelluloses and lignin with unaltered structures were prepared from control (CK) and C3H down-regulated 84K poplars. Modern analytical techniques, such as 13C NMR, 2D-HSQC NMR, and gel chromatography (GPC), were performed to better delineate the structural changes of hemicelluloses and lignin caused by transgenesis. Results showed that both the hemicelluloses (H-CK and H-C3H) extracted from control and C3H down-regulated poplar wood have a chain backbone of (1→4)-ß-D-Xylan with 4-O-Me-α-D-GlcpA as side chain, and the branch degree of the H-C3H is higher than that of H-CK. With regarding to the lignin macromolecules, NMR results demonstrated that the syringyl/guaiacyl (S/G) ratio and dominant substructure ß-O-4 linkages in C3H down-regulated poplar were lower than those of control poplar wood. By contrast, native lignin from C3H down-regulated poplar wood exhibited higher contents of p-hydroxybenzoate (PB) and p-hydroxyphenyl (H) units. In short, C3H down-regulation resulted in the chemical and structural changes of the hemicelluloses and lignin in these poplar wood. The identified structures will facilitate the downstream utilization and applications of lignocellulosic materials in the biorefinery strategy. Furthermore, this study could provide some illuminating results for genetic breeding on the improvement of wood properties and efficient utilization of poplar wood.

Short-time deep eutectic solvents pretreatment enhanced production of fermentable sugars and tailored lignin nanoparticles from abaca.

Deep eutectic solvents (DES) pretreatment is a promising approach to decrease "biomass recalcitrance" and boost the cellulose bioconversion as well as lignin valorization. In this study, a short-time DES pretreatment strategy was performed to enhance the production of high-yield fermentable sugars and tailored lignin nanoparticles (LNPs) from abaca. The glucose yield reached 92.4% under the optimal pretreatment condition (110 °C, 30 min), which was dramatically increased in comparison with that (9.5%) of control abaca. Simultaneously, nuclear magnetic resonance (NMR) and gel permeation chromatography (GPC) techniques indicated that the removed and regenerated DES lignin fractions displayed depolymerized structures and have relatively low molecular weight with relatively homogeneous morphology and narrow size distribution. Transmission electron microscope (TEM) analysis indicated that these lignin fractions are LNPs and the size of the optimal LNPs fraction is ranged from 30 nm to 50 nm. Moreover, all the DES lignin exhibited excellent antioxidant activities as compared to the commercial antioxidant butylated hydroxytoluene (BHT), which can be used as a promising natural antioxidant in industry. In short, this study demonstrated that the short-time DES pretreatment will improve the enzymatic digestibility and facilitate the controllable production and valorization of LNPs from abaca biomass, which will further promote the economic and overall benefits of biorefinery.

Retraction Note to: An assessment of transgenomics as a tool for gene discovery in Populus euphratica Oliv.

This article (Zhou et al. 2018) has been retracted by the authors because the sequence BIBAC 002A111F06 was incorrectly assigned to the wrong bacterial species. The BIBAC 002A111F06 sequence (GenBank Accession KC129717) reported in the paper was attributed to Populus euphratica Oliv. The BLAST search of this KC129717 sequence against the nr database at NCBI showed that it has very high similarity to a genomic sequence from the gram-negative bacteria Stenotrophomonas maltophilia. The bacterium associates with Populus euphratica Oliv. and DNA isolated from Populus euphratica Oliv. for the construction of the BIBAC clone library inlcuded DNA from Stenotrophomonas maltophilia. Therefore, the phenotype of the transgenic Arabidopsis line carrying the KC129717 sequence cannot be attributed to genes from Populus euphratica Oliv. The authors apologize for the confusion and misinterpretation of our data resulting from the incorrect sequence assignment. All authors agree to this retraction.

An assessment of transgenomics as a tool for gene discovery in Populus euphratica Oliv.

KEY MESSAGE: Transgenomics for gene discovery in Populus euphratica. Transgenomics, a member of the omics family of methodologies, is characterized as the introduction of DNA from one organism into another on a genome-wide scale followed by the identification of recipients with altered phenotypes. This strategy allows investigators to identify the gene(s) involved in these phenotypic changes. It is particularly promising for woody plants that have a long life cycle and for which molecular tools are limited. In this study, we constructed a large-insert binary bacterial artificial chromosome library of Populus euphratica, a stress-tolerant poplar species, which included 55,296 clones with average insert sizes of about 127 kb. To date, 1077 of the clones have been transformed into Arabidopsis thaliana via Agrobacterium by the floral dip method. Of these, 69 transgenic lines showed phenotypic changes represented by diverse aspects of plant form and development, 22 of which were reproducibly associated with the same phenotypic change. One of the clones conferring transgenic plants with increased salt tolerance, 002A1F06, was further analyzed and the 127,284 bp insert in this clone harbored eight genes that have been previously reported to be involved in stress resistance. This study demonstrates that transgenomics is useful in the study of functional genomics of woody plants and in the identification of novel gene(s) responsible for economically important traits. Thus, transgenomics can also be used for validation of quantitative trait loci mapped by molecular markers.

Effect of [Emim]Ac pretreatment on the structure and enzymatic hydrolysis of sugarcane bagasse cellulose.

Effect of ionic liquid pretreatment on enzymatic hydrolysis of cellulose was investigated in terms of the changes in the chemical and physical structure of the preparation. In this case, original cellulose isolated from sugarcane bagasse was subjected to ionic liquid ([Emim]Ac) dissolution at a mild temperature (90 °C) followed by regeneration in water and subsequently hydrolyzed by commercial cellulases. The original and regenerated cellulose were thoroughly characterized by XRD, FT-IR, CP/MAS (13)C NMR, and SEM. It was found that the original cellulose experienced an increase in glucose content from 80.0-83.3% to 91.6-92.8%, a decrease in the degree of polymerization from 974-1039 to 511-521, a crystal transformation from cellulose I to cellulose II, as well as an increase of surface area during the pretreatment. The results suggested that pretreatment led to effective disruption of cellulose for subsequent enzyme hydrolysis as evidenced by a high glucose conversion yield of 95.2%.

Autohydrolysis of bamboo (Dendrocalamus giganteus Munro) culm for the production of xylo-oligosaccharides.

Bamboo culm (Dendrocalamus giganteus Munro) was treated with hot water in non-isothermal conditions (140, 160, 180 and 200°C) for various reaction times (10, 30, 60 and 120 min) to generate xylo-oligosaccharides in a batch reactor at a solid to liquor ratio of 1:10 g/g. The influence of reaction conditions on yields, compositions and characteristics of the products were evaluated. The maximum yield of xylo-oligosaccharides of 47.49% was achieved at 180°C for 30 min with a relatively low level of xylose (4.73%) and other products (arabinose 0.20 g/L, galactose 0.19 g/L, HMF 0.15 g/L, furfural 0.36 g/L, acetic acid 0.64 g/L and formic acid 0.26 g/L). Higher temperature and/or longer reaction time reduced the yield of oligosaccharides, and enhanced concentrations of monosaccharides and byproducts. The result demonstrated that the yield of xylo-oligosaccharides obtained by autohydrolysis depends on the reaction temperature and time.

Structural features and antioxidant activity of xylooligosaccharides enzymatically produced from sugarcane bagasse.

Xylooligosaccharides (XOS) were prepared from xylan-rich hemicelluloses isolated by potassium hydroxide from sugarcane bagasse by hydrolysis with crude xylanase secreted by Pichia stipitis. Hydrolysis for 12h produced XOS with a maximum yield of 31.8%, equivalent to 5.29 mg mL(-1) in the hydrolyzate. XOS with degrees of polymerization (DP) from 2 to 4 (xylobiose, xylotriose, and xylotetraose) were the major components in the hydrolysates, whereas the oligosaccharides with higher DP of 5-6 (xylopentaose and xylohexose) showed a constant low level. FT-IR and NMR ((1)H, (13)C, HSQC) demonstrated that XOS contained Araf and 4-O-Me-α-D-GlcpA residues. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay showed that the XOS exhibited concentration-dependent antioxidant activity. The results obtained indicate that the XOS produced from sugarcane bagasse can be employed in food-related applications.

[The effect of different dose of verapamil and propranolol in kalium cardioplegia on the function of immature rat heart].

AIM: To explore the appropriate dose of the verapamil and propranolol in kalium cardiaplegia (KVP) by observation of the effect on the function of ischemic immature rat heart and compared with ST. Thomas II cardiaplegia. METHODS: 48 isolated hearts from Sprague-Dawley rats of 60 to approximately 80 g body weight, 22 +/- 2 days, male or female are perfused by Langendorff method for 20 min, and assigned to 1 of the following 6 groups (n = 8): control (CON), continuously perfused for 150 min. Ischemia/reperfusion (I/R), perfused with Locke's solution without glucose and oxygen equilibration for 3 min then no perfusion 27 min, repeated 3 cycles (ischemia for 90 min), followed by reperfusion for 60 min. Ischemia protected with ST. Thomas II cardioplegia (ST), each 3 min perfusion with ST. Thomas II cardioplegia during ischemia. Ischemia protected with three dose KVP cardioplegia (L, M, and H), perfused with ST. Thomas II cardioplegia containing verapamil and propranolol (x 10(-7) mol L(-1)) respectively 2.0, 0.34 (L), 6.8, 1.1 (M), 20,3.4 (H) during each 3 min perfusion of ischemia. Heart rate (min (-1), tens on(g), contraction force(g), peak systolic velocity (g.s-1), peak diastole velocity (g.s-), coronary flow (ml x min(-1 ), re-beat time (s) were monitored during the ischemia/ reperfusion. RESULTS: Compared to CON group, heart tension was rose when ischemia for 40 min and kept higher and could not rebeat after reperfusion in I/R group, In ST group, heart tension was rose after ischemia for 60 min and could re-beat but the pulse was weaker. Compared with ST group, KVP decreased the ischemic cardiac tension in dose dependently and the re-beat was stronger in L, M, and H groups. While compared with CON group, in L group, heart tension was rose when ischemia for 60 min and the re-beat was weaker. In H group, the heart tension was maintained lower when ischemia for 40 min and the re-beat was delay and weaker. Only in M group, heart tension was maintained stable during ischemia for 90 min and re-beat was stronger after reperfusion. CONCLUSION: Kalium cardiaplegia containing verapamil 6.8 x 10(-7) mol x L(-1) and propranolol 1.1 x 10(-7) mol x L(-1) has the best effect to protect the immature heart from ischemic injury.