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A new phenylethyl glycoside (1), (S)-1-phenylethyl-6-α-L-arabinopyranosyl-β-D-glucopyranoside), and a new dibenzocyclooctadiene lignan (7), Kasuracin A, together with seven known compounds, were isolated and identified from the leaves of Kadsura coccinea. Their structures were determined by analysis of multinuclear and multidimensional nuclear magnetic resonance (NMR), circular dichroism (CD), mass spectrometry, hydrolysis analysis, or comparison to those reported in the literature. Then, all the isolates 1‒9 were evaluated for their inhibitory effects against nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Among them, compounds 2 and 3 were isolated for the first time from this plant and compounds 3, 7, 8, and 9 significantly inhibited NO production with IC50 values of 36.4, 10.2, 12.3, and 21.7 μM, respectively.

We first confirmed the involvement of MalQ (4-α-glucanotransferase) in Escherichia coli glycogen breakdown by both in vitro and in vivo assays. In vivo tests of the knock-out mutant, ΔmalQ, showed that glycogen slowly decreased after the stationary phase compared to the wild-type strain, indicating the involvement of MalQ in glycogen degradation. In vitro assays incubated glycogen-mimic substrate, branched cyclodextrin (maltotetraosyl-β-CD: G4- β-CD) and glycogen phosphorylase (GlgP)-limit dextrin with a set of variable combinations of E. coli enzymes, including GlgX (debranching enzyme), MalP (maltodextrin phosphorylase), GlgP and MalQ. In the absence of GlgP, the reaction of MalP, GlgX and MalQ on substrates produced glucose-1-P (glc-1-P) 3-fold faster than without MalQ. The results revealed that MalQ led to disproportionate G4 released from GlgP-limit dextrin to another acceptor, G4, which is phosphorylated by MalP. In contrast, in the absence of MalP, the reaction of GlgX, GlgP and MalQ resulted in a 1.6-fold increased production of glc-1-P than without MalQ. The result indicated that the G4-branch chains of GlgP-limit dextrin are released by GlgX hydrolysis, and then MalQ transfers the resultant G4 either to another branch chain or another G4 that can immediately be phosphorylated into glc-1-P by GlgP. Thus, we propose a model of two possible MalQ-involved pathways in glycogen degradation. The operon structure of MalP-defecting enterobacteria strongly supports the involvement of MalQ and GlgP as alternative pathways in glycogen degradation.

Four new compounds N-salicyl-3-hydroxyanthranilic acid methyl ester (1), N-(2′-dehydroxysalicyl)-3-hydroxyanthranilic acid methyl ester (2), methyl-4-β-D-allopyranosyl-ferulate (3), and methyl-4-β-D-gulopyranosyl-cinnamate (4), along with six known compounds (5–10), were isolated from the roots of Aconitum carmichelii Debx. Their structures were elucidated on the basis of spectral data analysis, including 1D, 2D-NMR, and HR-ESI-MS. Compounds 1 and 2 showed the inhibition of nitric oxide (NO) production with IC50 values of 9.13 and 19.94 μM, respectively.

Findings on the decomposition of persistent luminescent (PeL) SrAl2O4:Eu2+,Dy3+ microparticles (MPs) in phosphate glass melt under static condition are reported. PeL phosphate glasses with the composition (50P2O5-10Na2O-40SrO) (in mol%) were prepared by adding the MPs in the glass melt. The decomposition of the MPs occurs during the preparation of the glass and leads to changes in the Eu2+ sites and to the formation of Eu3+ which decreases the PeL properties of the glasses. The decomposition of the MPs depends on the temperature at which the MPs are added in the melt and also on the time before casting the melts.

Overexpression of the flap endonuclease FEN1 has been observed in a variety of cancer types and is a marker for poor prognosis. To better understand the cellular consequences of FEN1 overexpression we utilized a model of its Saccharomyces cerevisiae homolog, RAD27. In this system, we discovered that flap endonuclease overexpression impedes replication fork progression and leads to an accumulation of cells in mid-S phase. This was accompanied by increased phosphorylation of the checkpoint kinase Rad53 and histone H2A-S129. RAD27 overexpressing cells were hypersensitive to treatment with DNA damaging agents, and defective in ubiquitinating the replication clamp proliferating cell nuclear antigen (PCNA) at lysine 164. These effects were reversed when the interaction between overexpressed Rad27 and PCNA was ablated, suggesting that the observed phenotypes were linked to problems in DNA replication. RAD27 overexpressing cells also exhibited an unexpected dependence on the SUMO ligases SIZ1 and MMS21 for viability. Importantly, we found that overexpression of FEN1 in human cells also led to phosphorylation of CHK1, CHK2, RPA32 and histone H2AX, all markers of genome instability. Our data indicate that flap endonuclease overexpression is a driver of genome instability in yeast and human cells that impairs DNA replication in a manner dependent on its interaction with PCNA.

In this study, the relationship between heavy metals (HMs) concentrations in soils and several weeds including Cyclosorus parasiticus, Dicranopteris linearis, Pityrogramma calomelanos, and Pteris vittata in three mining sites (Cam Gia (Thai Nguyen city), Tan Long (Dong Hy district), and Ha Thuong (Dai Tu district)) in Thai Nguyen province, Vietnam, have been investigated. The levels of HMs varied among soil origins and showed the contaminations of As, Cu, and Pb in soil samples collected in Dong Hy and Dai Tu districts. In addition, the HM distribution and cocontamination phenomena in different soils significantly affected the HM residues and transportation abilities into different species as well as tissues. Moreover, based on the analysis of bioaccumulation factor (BF) and translocation factor (TF), C. parasiticus and D. linearis were found potentially for phytoextraction by roots, while P. calomelanos and P. vittata were suitable for hyperaccumulation in shoots and leaves. Consequently, the strongest antioxidant property by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion (SRSA) radical scavenging assays were demonstrated in the methanol root extracts of C. parasiticus and P. vittata, respectively. In conclusion, the correlation among HM in soils and tissues with antioxidant property allows us to hypothesize that the presence of these elements can enhance the antioxidant activity of plant extracts, suggesting to apply the weeds in phytoremediation as well as in phytomedicine.

Marine microorganisms are an invaluable source of novel active secondary metabolites possessing various biological activities. In this study, the extraction and isolation of the marine sediment Penicillium species collected in Vietnam yielded ten secondary metabolites, including sporogen AO-1 (1), 3-indolecarbaldehyde (2), 2-[(5-methyl-1,4-dioxan-2-yl)methoxy]ethanol (3), 2-[(2R-hydroxypropanoyl)amino]benzamide (4), 4-hydroxybenzandehyde (5), chrysogine (6), 3-acetyl-4-hydroxycinnoline (7), acid 1H-indole-3-acetic (8), cyclo (Tyr-Trp) (9), and 2’,3’-dihydrosorbicillin (10). Their structures were identified by the analysis of 1D and 2D NMR data. Among the isolated compounds, 2-[(5-methyl-1,4-dioxan-2-yl)methoxy]ethanol (3) showed a strong inhibitory effect against Enterococcus faecalis with a minimum inhibitory concentration value of 32 µg/mL. Both 2-[(2R-hydroxypropanoyl)amino]benzamide (4) and 4-hydroxybenzandehyde (5) selectively inhibited E. coli with minimum inhibitory concentration values of 16 and 8 µg/mL, respectively. 2’,3’-Dihydrosorbicillin (10) potentially inhibited α-glucosidase activity at a concentration of 2.0 mM (66.31%).

Upconversion (UC) was obtained from phosphate glasses which contain only 0.01 at.% of Er3 + and 0.06 at.% of Yb3 +. The glasses were prepared using direct doping method by adding NaYF4:Er3 +, Yb3 + nanoparticles (NPs) into the glass at a doping temperature of 575 °C and 3 minute dwell time. The fraction of NP's survival during glass preparation can be augmented from 5 to 20% when replacing Na2O by NaF in the glass. Our study suggests that the corrosion behavior of the glass melt is an important parameter to consider in order to engineer new NPs-containing glasses with high UC efficiency.

Eurycoma longifolia is an herbal medicinal plant popularly used in Southeast Asian countries. In the present study, we show that 7-methoxy-(9H-β-carbolin-1-il)-(E)-1-propenoic acid (7-MCPA), a β-carboline alkaloid isolated from E. longifolia, exerted anti-inflammatory effects by activating the nuclear factor-E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. 7-MCPA inhibited lipopolysaccharide (LPS)-induced production of nitric oxide (NO), prostaglandin E2 (PGE2 ), and interleukin-6 (IL-6) in RAW264.7 cells and rescued C57BL/6 mice from LPS-induced lethality in vivo. LPS-induced expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and IL-6 was also significantly suppressed by treatment of 7-MCPA in RAW264.7 cells. 7-MCPA induced nuclear translocation of Nrf2 and increased transcription of its target genes, such as HO-1. Treating RAW264.7 cells with 7-MCPA increased the intracellular level of reactive oxygen species (ROS) and the phosphorylation level of p38 mitogen-activated protein kinase (MAPK); however, co-treatment with the antioxidant N-acetyl-cysteine (NAC) blocked 7-MCPA-induced p38 MAPK phosphorylation. Moreover, NAC or SB203580 (p38 MAPK inhibitor) blocked 7-MCPA-induced nuclear translocation of Nrf2, suggesting that 7-MCPA activated Nrf2 via a ROS-dependent p38 pathway. 7-MCPA induced HO-1 protein and mRNA expression and knockdown of Nrf2 with siRNA or SB203580 blocked 7-MCPA-mediated induction of HO-1 expression. Inhibiting Nrf2 or HO-1 abrogated the anti-inflammatory effects of 7-MCPA in LPS-stimulated RAW264.7 cells. We also demonstrated that 7-MCPA suppressed LPS-induced nuclear factor κB (NF-κB) activation. These results provide the first evidence that 7-MCPA exerts its anti-inflammatory effect by modulating the Nrf2 and NF-κB pathways and may be a potential Nrf2 activator to prevent or treat inflammatory diseases.

Jackfruit seed starch (JFSS) cultivated in Vietnam had a high amylose content (∼44%). Differential scanning calorimetry thermogram of JFSS consisted of two separate endothermic peaks with distinct onset and conclusion temperatures. When JFSS was heated at 70 °C, the first endothermic peak disappeared, whereas the second endotherm remained. The partially-gelatinized JFSS still possessed A-type X-ray diffraction pattern. SEM images showed that the rim of the starch granule at the hollow bottom of the bell shape first melted during the partial gelatinization at 70 °C. Both native and partially-gelatinized JFSS produced soft and highly elastic gels, but their moduli differed slightly. Additionally, the partially-gelatinized JFSS was successfully used to produce genistin–cycloamylose (CA) and amylose–lysolecithin complexes with and without 4-α-glucanotransferase. Water sorption isotherms of partially gelatinized JFSS showed higher water-holding capacity than that of the native starch.

Presence of β-amylase in ramie leaf and its anti-staling effect on starch-based foods were assessed. The ammonium sulfate fractionate (80% saturation) of the ramie leaf extracts showed a β-amylase activity, giving maltose (Glc2) as a major product, exclusively, when incubating with maltopentaose (Glc5) or soluble starch at 45 o C, pH 6.0. The starch-based food product (rice cake) prepared with freeze-dried ramie leaf enzyme revealed that the linear maltooligosaccharides ranging from Glc2 to Glc6 significantly increased and the shorter branch chains (DP 16) decreased in the product. These results demonstrated that maltosyl residue was released from the non-reducing end of the longer branch chains of amylopectin by β-amylase. The ramie leaf-treatment sample significantly reduced the retrogradation rate during 48 h storage at 4 o C. As an alternative plant-origin enzyme, the ramie leaf β-amylase has potential for a novel anti-staling additive.

SLX4, a coordinator of multiple DNA structure-specific endonucleases, is important for several DNA repair pathways. Non-covalent interactions of SLX4 with ubiquitin are required for localizing SLX4 to DNA-interstrand crosslinks (ICLs), yet how SLX4 is targeted to other functional contexts remains unclear. Here, we show that SLX4 binds SUMO-2/3 chains via SUMO-interacting motifs (SIMs). The SIMs of SLX4 are dispensable for ICL repair, but important for processing CPT-induced replication intermediates, suppressing fragile site instability, and localizing SLX4 to ALT telomeres. The localization of SLX4 to laser-induced DNA damage also requires the SIMs, as well as DNA-end resection, UBC9 and MDC1. Furthermore, the SUMO binding of SLX4 enhances its interaction with specific DNA-damage sensors or telomere-binding proteins, including RPA, MRE11-RAD50-NBS1 and TRF2. Thus, the interactions of SLX4 with SUMO and ubiquitin increase its affinity for factors recognizing different DNA lesions or telomeres, helping to direct the SLX4 complex in distinct functional contexts.

Heterozygous somatic mutations in spliceosome genes (U2AF1, SF3B1, ZRSR2, or SRSF2) occur in >50% of patients with myelodysplastic syndrome (MDS). These mutations occur early in disease development, suggesting that they contribute to MDS pathogenesis and may represent a unique genetic vulnerability for targeted therapy. Here, we show that RNA splicing perturbation by expression of the U2AF1(S34F) mutant causes accumulation of R loops, a transcription intermediate containing RNA:DNA hybrids and displaced single-stranded DNA, and elicits an ATR response. ATR inhibitors (ATRi) induced DNA damage and cell death in U2AF1(S34F)-expressing cells, and these effects of ATRi were enhanced by splicing modulating compounds. Moreover, ATRi-induced DNA damage was suppressed by overexpression of RNaseH1, an enzyme that specifically removes the RNA in RNA:DNA hybrids, suggesting that the ATRi sensitivity of U2AF1(S34F)-expressing cells arises from R loops. Taken together, our results demonstrate that ATR may represent a novel therapeutic target in patients with MDS carrying the U2AF1(S34F) mutation and potentially other malignancies harboring spliceosome mutations. Significance: This study provides preclinical evidence that patients with MDS or other myeloid malignancies driven by spliceosome mutations may benefit from ATR inhibition to exploit the R loop–associated vulnerability induced by perturbations in splicing. Cancer Res; 78(18); 5363–74. ©2018 AACR.

In this invited proceeding, we report our latest results on the development of Er3+ doped phosphate glass-ceramics. Those new glass-ceramics were processed using different techniques, such as direct doping of particles into the glass melt and glass-ceramics methods. First, we explain the challenges to balance the survival and dispersion of Er3+ doped particles when preparing phosphate glass-ceramics using the direct doping of particles. Then, we discuss the impact of the glass crystallization on the luminescence properties of Er3+ ions.

Thermostable disproportionation enzymes active at around the gelatinization temperature of rice starch (70–80 °C) were used to improve the quality of cooked rice with lump-free properties. Cyclodextrin glucanotransferase from Pyrococcus furiosus (CGTase) and a starch binding domain-attached alpha-glucanotransferase from Thermus aquaticus (αGTase) were prepared from an Escherichia coli transformant carrying a recombinant plasmid. The stickiness determined by a texture analyzer showed that both enzymes had the effect of reducing the lumping properties of cooked rice grains and rice porridge. The enzyme treatment also provided a finer, creamier, and more uniform texture of rice porridge. An HPAEC analysis of the starch structure in cooked rice and rice porridge revealed that the proportion of both short (DP 20) increased in the enzyme-treated samples. Additionally, the amylose content of the cooked rice grain treated with αGTase and CGTase decreased by 36% and 30%, whereas that of porridge was reduced by 36% and 35%, respectively. In particular, the amylose content on the surface of the enzyme-treated cooked rice grains was reduced by 50% compared with that of untreated cooked rice. These results indicated that the glucan segment of amylose was transferred to amylopectin branch chains by a transferring reaction with both enzymes, reducing the amylose content of grains during cooking. Mechanistically, the decrease in the amylose content filmed on the surface is likely to be responsible for reducing the stickiness of cooked rice grains and rice porridge. Furthermore, the retrogradation rate determined by a differential scanning calorimeter was significantly reduced in the enzyme-treated samples after storage for 10 h at 4 °C. The decrease in long-chain amyloses, may lead to the retardation of re-crystallization. These physicochemical properties of the modified cooked rice and rice porridge can be applied to various rice-based products in the food industry.

The O-demethylases of anaerobes are corrinoid-dependent, ether-cleaving methyltransferase enzyme systems consisting of four components. The interaction of the O-demethylase components of the acetogenic bacterium Acetobacterium dehalogenans was studied by protein mobility on native PAGE, far-Western blot analysis and yeast two-hybrid screen. Using native PAGE and far-Western blot, the interaction of the activating enzyme (AE) with its substrate, the corrinoid protein (CP), could be observed. The interaction occurred with four different CPs of A. dehalogenans and a CP from Desulfitobacterium hafniense DCB-2, all involved in ether cleavage. In the corrinoid reduction assay, the AE reduced all CPs tested. This result indicates a broad substrate specificity of the AE of A. dehalogenans. In addition, an interaction of the A. dehalogenans CP of the vanillate-O-demethylase with the two methyltransferases of the same enzyme system was observed. The interaction of the ether-cleaving methyltransferase with the CP appeared to be significantly less pronounced than that reported for the homologous methanol and methylamine methyltransferase systems of methanogenic archaea.

SummaryLoss of minichromosome maintenance protein 10 (Mcm10) causes replication stress. We uncovered that S. cerevisiae mcm10-1 mutants rely on the E3 SUMO ligase Mms21 and the SUMO-targeted ubiquitin ligase complex Slx5/8 for survival. Using quantitative mass spectrometry, we identified changes in the SUMO proteome of mcm10-1 mutants and revealed candidates regulated by Slx5/8. Such candidates included subunits of the chromosome passenger complex (CPC), Bir1 and Sli15, known to facilitate spindle assembly checkpoint (SAC) activation. We show here that Slx5 counteracts SAC activation in mcm10-1 mutants under conditions of moderate replication stress. This coincides with the proteasomal degradation of sumoylated Bir1. Importantly, Slx5-dependent mitotic relief was triggered not only by Mcm10 deficiency but also by treatment with low doses of the alkylating drug methyl methanesulfonate. Based on these findings, we propose a model in which Slx5/8 allows for passage through mitosis when replication stress is tolerable.

Caesalpinia sappan Linn. (Leguminosae) has been used in folk medicines for the treatment of many diseases. The heartwood of this plant contains various phenolic components with interesting biological applications; however, the chemical and biological potentials of the seed of this plant have not been fully explored.This study identified the cytotoxic activity of compounds from the seeds of C. sappan.The methanol extract of the seed of C. sappan was suspended in H2O and then partitioned with CH2Cl2, EtOAc, and n-BuOH, successively. Diterpenoid compounds were isolated from the CH2Cl2-soluble fraction by silica gel column chromatography methods using organic solvents. The compound structures were determined by detailed analysis of NMR and MS spectral data. Cytotoxic activity was measured using a modified MTT assay against HL-60, HeLa, MCF-7, and LLC cancer cells. The activation of caspase-3 enzyme and western blotting assay were performed to confirm inhibitory mechanism of active compound.Five cassane-type diterpenoids were isolated and identified as phanginin I (1), phaginin A (2), phanginin D (3), phanginin H (4), and phanginin J (5). Compounds 1-4 showed effective inhibition against HL-60 cells with the IC50 values of 16.4 ± 1.5, 19.2 ± 2.0, 11.7 ± 1.6, and 22.5 ± 5.1 μM. Compounds 1-3 exhibited cytotoxic activity against HeLa cells with the IC50 values of 28.1 ± 3.6, 37.2 ± 3.4, and 22.7 ± 2.8 μM. Treatment of HL-60 cell lines with various concentrations of 3 (0-30 μM) resulted in the growth inhibition and induction of apoptosis.These findings demonstrate that compound 3 (phanginin D) is one of the main active components of the seed of C. sappan activating caspases-3 which contribute to apoptotic cell death.

One new β-carboline alkaloid 7-methoxy-(9H-β-carbolin-1-il)-(E)-1-propenoic acid (1) together with 9-methoxycanthin-6-one (2) and 9-hydroxycanthin-6-one (3) were isolated from the hairy-root cultures of Eurycoma longifolia. The effects of these compounds on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 cells were investigated. Compound 1 strongly inhibited the production of NO while 2 and 3 having weak or inactive effect. Consistently, compound 1 decreased the expression of cyclooxygenase-2 and inducible nitric oxide synthase.

R loop, a transcription intermediate containing RNA:DNA hybrids and displaced single-stranded DNA (ssDNA), has emerged as a major source of genomic instability. RNaseH1, which cleaves the RNA in RNA:DNA hybrids, plays an important role in R loop suppression. Here, we show that replication protein A (RPA), a ssDNA-binding protein, interacts with RNaseH1 and colocalizes with both RNaseH1 and R loops in cells. In vitro, purified RPA directly enhances the association of RNaseH1 with RNA:DNA hybrids and stimulates the activity of RNaseH1 on R loops. An RPA binding-defective RNaseH1 mutant is not efficiently stimulated by RPA in vitro, fails to accumulate at R loops in cells, and loses the ability to suppress R loops and associated genomic instability. Thus, in addition to sensing DNA damage and replication stress, RPA is a sensor of R loops and a regulator of RNaseH1, extending the versatile role of RPA in suppression of genomic instability.

We studied the activity of a debranching enzyme (TreX) from Sulfolobus solfataricus on glycogen-mimic substrates, branched maltotetraosyl-β-cyclodextrin (Glc 4 -β-CD), and natural glycogen to better understand substrate transglycosylation and the effect thereof on glycogen debranching in microorganisms. The validation test of Glc 4 -β-CD as a glycogen mimic substrate showed that it followed the breakdown process of the well-known yeast and rat liver extract. TreX catalyzed both hydrolysis of α-1,6-glycosidic linkages and transglycosylation at relatively high (>0.5 mM) substrate concentrations. TreX transferred maltotetraosyl moieties from the donor substrate to acceptor molecules, resulting in the formation of two positional isomers of dimaltotetraosyl-α-1,6-β-cyclodextrin [(Glc 4 ) 2 -β-CD]; these were 6 1 ,6 3 - and 6 1 ,6 4 -dimaltotetraosyl-α-1,6-β-CD. Use of a modified Michaelis-Menten equation to study substrate transglycosylation revealed that the k cat and K m values for transglycosylation were 1.78 × 10 3 s −1 and 3.30 mM, respectively, whereas the values for hydrolysis were 2.57 × 10 3 s −1 and 0.206 mM, respectively. Also, enzyme catalytic efficiency (the k cat / K m ratio) increased as the degree of polymerization of branch chains rose. In the model reaction system of Escherichia coli, glucose-1-phosphate production from glycogen by the glycogen phosphorylase was elevated ∼1.45-fold in the presence of TreX compared to that produced in the absence of TreX. The results suggest that outward shifting of glycogen branch chains via transglycosylation increases the number of exposed chains susceptible to phosphorylase action. We developed a model of the glycogen breakdown process featuring both hydrolysis and transglycosylation catalyzed by the debranching enzyme.

Mutants with deletion mutations in the glg and mal gene clusters of Escherichia coli MC4100 were used to gain insight into glycogen and maltodextrin metabolism. Glycogen content, molecular mass, and branch chain distribution were analyzed in the wild type and in Δ malP (encoding maltodextrin phosphorylase), Δ malQ (encoding amylomaltase), Δ glgA (encoding glycogen synthase), and Δ glgA Δ malP derivatives. The wild type showed increasing amounts of glycogen when grown on glucose, maltose, or maltodextrin. When strains were grown on maltose, the glycogen content was 20 times higher in the Δ malP strain (0.97 mg/mg protein) than in the wild type (0.05 mg/mg protein). When strains were grown on glucose, the Δ malP strain and the wild type had similar glycogen contents (0.04 mg/mg and 0.03 mg/mg protein, respectively). The Δ malQ mutant did not grow on maltose but showed wild-type amounts of glycogen when grown on glucose, demonstrating the exclusive function of GlgA for glycogen synthesis in the absence of maltose metabolism. No glycogen was found in the Δ glgA and Δ glgA Δ malP strains grown on glucose, but substantial amounts (0.18 and 1.0 mg/mg protein, respectively) were found when they were grown on maltodextrin. This demonstrates that the action of MalQ on maltose or maltodextrin can lead to the formation of glycogen and that MalP controls (inhibits) this pathway. In vitro , MalQ in the presence of GlgB (a branching enzyme) was able to form glycogen from maltose or linear maltodextrins. We propose a model of maltodextrin utilization for the formation of glycogen in the absence of glycogen synthase.