Search

The characterization of pore systems in fine-grained carbonate rocks faces numerous challenges due to the significant complexity of microscopic features, including a variation of micro- and nanoscale pore sizes and the complex pore-throat distribution. In this work, digital core technology was adopted to characterize the pore systems of lacustrine fine-grained carbonate rocks in the Yingxi area of Qaidam Basin. The simulated results indicated that the pore types predominantly contain intercrystalline and dissolution pores. The former exhibit high porosity but extremely low permeability and are primarily developed in bedded dolostones. Conversely, the latter show relatively higher permeability, predominantly developed in bedded calcareous dolostones. The elevated dolomite content provides the material basis for the development of intercrystalline pores, while the extremely small throat radius constrains the fluidity of this pore system. In addition, the dissolution has a great impact on improving the permeable capability of intercrystalline pore system via increasing the radius and specific surface area of pores and throats. [ABSTRACT FROM AUTHOR]

The time-dependent wave packet method has been employed to calculate state-to-state differential cross sections for the title reaction in full dimensions. It is found that the majority of H2O is produced in the first stretching excited states, with a large fraction of available energy for the reaction ending up as product internal motion. The differential cross sections for collision energy up to 0.4 eV are all peaked in the backward direction, but the width of the angular distribution increases considerably as the increase of collision energy. The isotope effect was also examined by comparing the scattering angular distribution for the title reaction with those for the HD + OH and D2 + OH reactions obtained in our previous work. [ABSTRACT FROM AUTHOR]

Full quantum mechanical calculations of vibrational energies of methane and fluoromethane are carried out using a polyspherical description combining Radau and Jacobi coordinates. The Hamiltonian is built in a potential-optimized discrete variable representation, and vibrational energies are solved using an iterative eigensolver. This new approach can be applied to a large variety of molecules. In particular, we show that it is able to accurately and efficiently compute eigenstates for four different molecules : CH4, CHD3, CH2D2, and CH3F. Very good agreement is obtained with the results reported previously in the literature with different approaches and with experimental data. [ABSTRACT FROM AUTHOR]

Pancreatic cancer has a low survival rate globally. Anillin (ANLN) is involved in the pathogenesis of pancreatic cancer (PC). The present study used databases and reverse transcription-quantitative PCR to investigate the association between ANLN expression, clinical variables and the survival rate of patients with pancreatic cancer. Gene expression of ANLN in normal and cancer tissues was analyzed using data from The Cancer Genome Atlas, Oncomine and Gene Expression database of Normal and Tumor tissues 2 and ANOVA, and the association between ANLN mRNA expression and ANLN genovariation was analyzed using cBioPortal. The association between ANLN expression and the survival, clinical, pathological and prognostic characteristics of PC was analyzed using Kaplan-Meier (K-M) survival analysis, Kruskal Wallis and Mann Whitney-U tests, and logistic and Cox regression models. Gene Set Enrichment Analysis (GSEA) revealed the molecular pathways underpinning ANLN function in PC. Overexpression of ANLN was observed in PC cells (normal vs. tumor, P<0.01) and tissues (normal vs. tumor, P=0.008). Enhanced ANLN expression was associated with high tumor grade (grade 1 vs. grade 3, odds ratio: 5.662, P<0.001). However, ANLN expression was not associated with other clinical features (all P>0.05). K-M analysis suggested that increased ANLN expression was associated with poor survival (P=0.002). Univariate and multivariate analysis revealed the ANLN is an independent prognostic factor for PC (P<0.001). GSEA demonstrated the p53, cell cycle, DNA replication, mismatch repair, nucleotide excision repair and PC pathways were associated with low expression of ANLN. Overall, ANLN is more highly expressed in PC compared with in normal tissue, and is associated with poor differentiation. The expression of ANLN may be a novel prognostic marker of poor survival. Finally, ANLN exert its functions in PC through the p53, cell cycle, DNA replication, mismatch repair and nucleotide excision repair and pathways. [ABSTRACT FROM AUTHOR]

In agricultural production, the main cause of groundwater pollution has been the leakage of total Nitrogen (TN) caused by excessive water and fertilizer application. In this study, an experiment combining a field and indoors was used to seek optimal coupling scheme and prevent groundwater pollution caused by excessive fertilization. Results showed that in recent years, groundwater quality has been deteriorating, and nitrate levels have increased by approximately three times (2014-2016). The TN accumulated significantly in a different soil layer, along with crop growth. The N accumulated mainly in the bottom of the studied soil layer (80-160 cm) over time. Under different water fertilizer treatments, the crop yield, leakage, and N use efficiency (NUE) were obviously various. The effect of irrigation water quantity was more significant for N leakage than fertilizer (P<0.05), and more water resulted in more N leakage. Additionally, considering the comprehensive benefits for winter wheat, medium fertilizer (N, 225 kg ha-1) is an optimal scheme with medium water (3150 m3 ha-1). For summer maize, medium water (1350 m3 ha-1) and high fertilizer (N, 162 m3 ha-1) were optimal treatment. The above schemes have the best comprehensive benefits for agricultural production in the case area. [ABSTRACT FROM AUTHOR]

Covalent modifications of DNA and histones are important epigenetic events and the genomewide reshaping of epigenetic markers is common in cancer. Epigenetic markers are produced by enzymatic reactions, and some of these reactions require the presence of metabolites, specifically Epigenetic Enzyme Required Metabolites (EERMs), as cofactors. Recent studies found that the abundance of these EERMs correlates with epigenetic enzyme activities. Also, the subcellular compartmentation, especially the nuclear localization of these EERMs, may play a role in regulating the activities of epigenetic enzymes. Moreover, genespecific recruitment of enzymes that produce the EERMs in the proximity of the epigenetic modification events accompanying the regulation of gene expression, were proposed. Therefore, it is important to summarize findings of EERMs in regulating epigenetic modifications at both the DNA and histone levels, and to understand how EERMs contribute to cancer development by addressing their global versus local distribution. [ABSTRACT FROM AUTHOR]

Objective: Ionic remodeling has a close correlation with the occurrence of atrial fibrillation (AF). Atrial tachypacing remodeling is associated with characteristic ionic remodeling. The purpose of this study was to assess the efficacy of cilostazol, an oral phosphodiesterase 3 inhibitor, for preventing atrial ionic remodeling in long-term rapid atrial pacing (RAP) dogs. Methods: We use the methods of patch-clamp and molecular biology to investigate the effect of cilostazol on ion channel and channel gene expression in long-term RAP dogs. Twenty-one dogs were randomly assigned to sham, control paced, and paced+cilostazol (5 mg/kg/d, cilo) groups, with 7 dogs in each group. The sham group was instrumented with a pacemaker but without pacing. RAP at 500 beats/min was maintained for 2 weeks in the paced and cilo groups. During the pacing, cilostazol was given orally in the cilo group. Whole-cell patch-clamp technique was used to record atrial L-type Ca2+ (ICaL) and fast sodium channel (INa) ionic currents. Western blot and RT-PCR were applied to estimate the gene expression of the ICaLα) 1C (Cav1.2) and INav1.5α) Nav1.5α) subunits. StatistICaL analysis was performed using SPSS 13.0. Results: The density of ICaL and INa currents (pA/pF) was significantly reduced in the paced group (ICaL: -6.55±1.42 vs. -4.46±0.59 pA/pF; INa: -48.24±10.54 vs. -30.48±5.20 pA/pF, p<0.01). The paced+cilo group could not increase the density of ICaL currents (ICaL: -4.37±1.25 pA/pF, p>0.05), while the INa currents were recovered (-44.54±12.65 pA/pF, p<0.01) compared with the paced group. The mRNA and protein expression levels of Cav1.2 and Nav1.5α were apparently down-regulated in the paced group (p<0.01), but after cilostazol treatment, both of these subunits were up-regulated significantly (p<0.01). Conclusion: Cilostazol may have protective effects on RAP-induced atrial ionic remodeling. [ABSTRACT FROM AUTHOR]

In the present paper, we describedthe design, synthesis, SAR,and biological profile of a novel spirocyclic sulfone series of γ-secretaseinhibitors (GSIs) related to MRK-560. We utilized an additional spirocyclicring system to stabilize the active chair conformation of the parentγ-secretase inhibitors. The resulting series is devoid of theCYP2C9 inhibition liability of MRK-560. A few representative analogswere assessed in a nontransgenic animal model of Alzheimer’sdisease (AD), demonstrating reduction of amyloid-β (Aβ)in the CNS after acute oral dosing. A spirocyclic phosphonate wasidentified as the optimal ring system for both potency and pharmacokinetics.Compared to GSIs studied in the clinic, representative spirocyclicphosphonate 18a(−)features improved selectivityfor the inhibition of the PS-1 isoform of γ-secretase (33-foldvs PS-2), which may alleviate the adverse effect profile of the clinicalGSIs. [ABSTRACT FROM AUTHOR]

A coupling process of anaerobic methanogenesis and electromethanogenesis was proposed to treat high organic load rate (OLR) wastewater. During the start-up stage, acetate removal efficiency of the electric-biological reactor (R1) reached the maximization about 19 percentage points higher than that of the control anaerobic reactor without electrodes (R2), and CH4 production rate of R1 also increased about 24.9% at the same time, while additional electric input was 1/1.17 of the extra obtained energy from methane. Coulombic efficiency and current recorded showed that anodic oxidation contributed a dominant part in degrading acetate when the metabolism of methanogens was low during the start-up stage. Along with prolonging operating time, aceticlastic methanogenesis gradually replaced anodic oxidation to become the main pathway of degrading acetate. When the methanogens were inhibited under the acidic conditions, anodic oxidation began to become the main pathway of acetate decomposition again, which ensured the reactor to maintain a stable performance. FISH analysis confirmed that the electric field imposed could enrich the H2/H+-utilizing methanogens around the cathode to help for reducing the acidity. This study demonstrated that an anaerobic digester with a pair of electrodes inserted to form a coupling system could enhance methanogenesis and reduce adverse impacts. [ABSTRACT FROM AUTHOR]

Research has shown that the palmitoyl group of α-tubulin mediates the hydrophobic interaction between microtubules and intracellular membranes and that palmitoylated tubulin plays a role in signal transduction. There are 20 cysteine residues per α/β tubulin heterodimer. C376 of α-tubulin was reported to be predominantly palmitoylated and C20, C213 and C305 of α-tubulin were palmitoylated at lower levels. The previous method used for the analysis of the palmitoylation sites on α-tubulin was based on H-labeling, enzymolysis, purification and sequencing. This approach, although efficient, is laborious. Mass spectrometry (MS), especially tandem MS, has been shown to be a successful method for identification of various post-translational modifications of proteins. We report here a convenient MS-based method to comprehensively analyze the palmitoylation sites of the α/β tubulin heterodimer. Acyl-biotinyl exchange chemistry and streptavidin agarose affinity purification were applied to enrich palmitoylated peptides from tubulin. After nano-LC-MS/MS analysis, database searching and manual analysis of the spectra revealed that 11 cysteine residues of the α/β tubulin heterodimer were palmitoylated. [ABSTRACT FROM AUTHOR]

Endothelialization repairs the lining of damaged vasculature and is a key process in preventing thrombosis and restenosis. It has been demonstrated that extracellular calcium ([Ca2+]o) influx is important for subsequent endothelialization. The role of intracellular Ca2+ stores in mechanical denudation induced intracellular calcium ([Ca2+]i) rise and endothelialization remains to be demonstrated. Using monolayer culture of a human endothelial cell line (human umbilical vein endothelial cell, HUVEC), we investigated [Ca2+]i wave propagation and re-endothelialization following mechanical denudation. Consistent with previous reports for other types of cells, mechanical denudation induces calcium influx, which is essential for [Ca2+]i rise and endothelialization. Moreover, we found that intracellular Ca2+ stores are also essential for denudation induced [Ca2+]i wave initiation and propagation, and the subsequent endothelialization. Thapsigargin which depletes intracellular Ca2+ stores completely abolished [Ca2+]i wave generation and endothelialization. Xestospongin C (XeC), which prevents Ca2+ release from intracellular Ca2+ stores by inhibition of inositol 1,4,5-trisphosphate (IP3) receptor, inhibited intercellular Ca2+ wave generation and endothelialization following denudation. Purinergic signaling through a suramin sensitive mechanism and gap junction communication also contribute to in intercellular Ca2+ wave propagation and re-endothelialization. We conclude that intracellular Ca2+ stores, in addition to extracellular Ca2+, are essential for intracellular Ca2+ signaling and subsequent endothelialization following mechanical denudation. J. Cell. Physiol. 214: 595–603, 2008. © 2007 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

A novel methodology for parallel liquid-phase synthesis of carbamates suitable for the preparation of sterically hindered molecules is disclosed. The alcohols are converted to 4-nitrophenylcarbonates, followed by the reaction with amines. Side product 4-nitrophenol and the unreacted excess amines are scavenged by appropriately chosen cleanup resins, selected among Amberlyst A26 (hydroxide form) and macroporous sulfonic acid (MP-TsOH) or polystyrene isocyanate (PS-NCO) and polystyrene benzaldehyde (PS-PhCHO) resins. As a part of a medicinal chemistry program directed toward finding γ-secretase inhibitors as prospective drug candidates for Alzheimer’s disease, a 6 × 24 library of carbamates was prepared. Out of 144 library members, 133 had a purity for the targeted compound of 80% or better. The prepared compounds were assessed in the γ-secretase inhibition assay and demonstrated activity with IC 50values in the range from 1 µM to 5 nM, with the activity of 7 compounds being better than 10 nM. [ABSTRACT FROM AUTHOR]

Determination of partial discharge locations in large oil-filled power transformers is unreliable and difficult using external acoustic sensors, and internal sensors would need to be non-metallic. Thus the use of a fibre-optic coil as an acoustic sensor would allow the detection and location of partial discharges with potentially a much greater sensitivity. This paper describes the design of such a sensor and its signal processing requirements, its construction and its initial testing. The sensor was found to be able to detect 1-Pa acoustic pressure waves under optimum conditions; and to detect partial discharges in small artificial voids in an oil-bath. [Copyright &y& Elsevier]

Determines the reaction mechanism of lactams with nucleophiles. Derivation of nucleophiles with butylaziridinone; Protonation of the lactam; Nucleophilicity of amines.

The conventional college snow and ice teaching multi-source resource intelligent management system has the problem of incomplete software resource analysis function, which leads to high CPU usage of system. A cloud platform based college snow and ice teaching multi-source resource intelligent management system is designed. Hardware part: record the state of the pins when the register is reset, account for the data bit width and storage capacity of DDR2 SDRAM memory, and optimise the data storage module; Part of the software: obtain ice and snow sports course objectives in colleges and universities, rationally organise the course organization form, optimise the intelligent management mode of teaching multi-source resources by using the cloud platform, support online browsing of various text resources, set relevant parameters to construct various random modification operations, and design the software resource analysis function with the knowledge fusion algorithm. Experimental results: The average CPU usage of the multi-source intelligent resource management system for snow and ice teaching in colleges and universities in this paper and the other two systems is 34.257%, 47.458%, 53.578%. Experimental results show that the performance of multi-source intelligent resource management system for snow and ice teaching in colleges and universities has been significantly improved after making full use of cloud platform. [ABSTRACT FROM AUTHOR]

Conventional martial arts teaching system has the problem that the interactive teaching mode is not three-dimensional enough, which leads to the slow byte sending speed of the system. In this study, a martial arts teaching system based on virtual reality technology is designed. In the hardware part, 5 V and 3.3 V DC regulated power supply is selected and Ethernet interface and USB for network connection are used. For the software part, according to the different degrees of muscle participation in the completion of the action, the technical characteristics of martial arts action are identified, virtual reality technology is used to build interactive teaching mode, the feedback of proprioceptors is used to adjust martial arts action, the software priority processing function is designed, and the priority of martial arts action frame is improved. The average sending byte speed of the martial arts teaching system in this paper and the other two martial arts teaching systems is 81.391 kb/s, 93.465 kb/s, and 94.557 kb/s, respectively, indicating that the performance of the martial arts teaching system has been improved after the combination of virtual reality technology. [ABSTRACT FROM AUTHOR]

A more flexible neural network (NN) method using the fundamental invariants (FIs) as the input vector is proposed in the construction of potential energy surfaces for molecular systems involving identical atoms. Mathematically, FIs finitely generate the permutation invariant polynomial (PIP) ring. In combination with NN, fundamental invariant neural network (FI-NN) can approximate any function to arbitrary accuracy. Because FI-NN minimizes the size of input permutation invariant polynomials, it can efficiently reduce the evaluation time of potential energy, in particular for polyatomic systems. In this work, we provide the FIs for all possible molecular systems up to five atoms. Potential energy surfaces for OH3 and CH4 were constructed with FI-NN, with the accuracy confirmed by full-dimensional quantum dynamic scattering and bound state calculations. [ABSTRACT FROM AUTHOR]

We report a permutationally invariant global potential energy surface (PES) for the H + CH4 system based on ~63 000 data points calculated at a high ab initio level (UCCSD(T)-F12a/AVTZ) using the recently proposed permutation invariant polynomial-neural network method. The small fitting error (5.1 meV) indicates a faithful representation of the ab initio points over a large configuration space. The rate coefficients calculated on the PES using tunneling corrected transition-state theory and quasi-classical trajectory are found to agree well with the available experimental and previous quantum dynamical results. The calculated total reaction probabilities (Jtot = 0) including the abstraction and exchange channels using the new potential by a reduced dimensional quantum dynamic method are essentially the same as those on the Xu-Chen-Zhang PES. [ABSTRACT FROM AUTHOR]

The present study investigated the correlations of the Tspan-1 gene expression with the clinical characteristics and survival prognoses of patients with advanced gastric cancer. A total of 150 patients with advanced gastric cancer were enrolled in the present study, of whom 84 were at stage II and 66 were at stage III according to the tumor node metastasis (TNM) staging; the immunohistochemical staining method and the semi-quantitative PCR method were used to detect the positive expression rates and mRNA relative expression levels of Tspan-1, vascular endothelial growth factor (VEGF), E-cadherin and N-cadherin. The positive expression rates of Tspan-1, VEGF, E-cadherin and N-cadherin were 58.0% (87 patients), 50.0% (75 patients), 28.0% (42 patients) and 53.3% (80 patients), respectively. The positive expressions and mRNA levels of Tspan-1, VEGF, E-cadherin and N-cadherin were not correlated with sex or age (P>0.05), but associated with the cancer state (stage II or stage III) and maximum tumor diameter (P<0.05). With the increase of stage and tumor diameter, the positive rates and mRNA levels of Tspan-1, VEGF and N-cadherin were increased, while those of E-cadherin were decreased. Among patients with stage II/III advanced gastric cancer, those with positive expression of Tspan-1, VEGF and N-cadherin had lower median survival time and survival rates than patients with negative expressions, while patients with positive expression of E-cadherin had higher median survival time and survival rate than those with negative expression (P<0.05). The high expression of Tspan-1 gene is associated with the TNM staging of advanced gastric cancer and the tumor diameter, influences the survival prognosis, and may involve the processes of angiogenesis and epithelial-mesenchymal transition. [ABSTRACT FROM AUTHOR]

This paper tested the permeability and porosity of oil shale during the pyrolysis process. Combined with the in-situ pyrolysis simulation, this paper analyzed and discussed the evolution of permeability and porosity in the oil shale in-situ pyrolysis zone through electric heating technology. During the pyrolysis of oil shale, the porosity and permeability increased significantly when temperature ranged from 300°C to 500°C. The scope of the in-situ pyrolysis zone expanded with the increase of heating time and pyrolysis saturation was reached essentially after six years of heating. After in-situ pyrolysis, oil shale's permeability and porosity changed greatly within 7.5 meters in the vertical direction and 4.8 meters in the horizontal direction from the central wellbore of the heating well. Oil shale in this area changed from aquifuge to aquitard. [ABSTRACT FROM AUTHOR]

PCBP2 is a member of the poly(C)-binding protein (PCBP) family, which plays an important role in posttranscriptional and translational regulation by interacting with single-stranded poly(C) motifs in target mRNAs. Several PCBP family members have been reported to be involved in human malignancies. Here, we show that PCBP2 is upregulated in human glioma tissues and cell lines. Knockdown of PCBP2 inhibited glioma growth in vitro and in vivo through inhibition of cell-cycle progression and induction of caspase-3-mediated apoptosis. Thirty-five mRNAs were identified as putative PCBP2 targets/interactors using RIP-ChIP protein-RNA interaction arrays in a human glioma cell line, T98G. Four-and-a-half LIM domain 3 (FHL3) mRNA was downregulated in human gliomas and was identified as a PCBP2 target. Knockdown of PCBP2 enhanced the expression of FHL3 by stabilizing its mRNA. Overexpression of FHL3 attenuated cell growth and induced apoptosis. This study establishes a link between PCBP2 and FHL3 proteins and identifies a new pathway for regulating glioma progression. [ABSTRACT FROM AUTHOR]

Background: D-Lactate is a valued chemical which can be produced by some bacteria including Klebsiella pneumoniae. However, only a few studies have focused on K. pneumoniae for D-lactate production with a significant amount of by-products, which complicated the purification process and decreased the yield of D-lactate. Results: Based on the redirection of carbon towards by-product formation, the effects of single-gene and multiplegene deletions in K. pneumoniae on d-lactate production from glucose via acetolactate synthase (budB), acetate kinase (ackA) and alcohol dehydrogenase (adhE) were tested. Klebsiella pneumoniae mutants had different production behaviours. The accumulation of the main by-products was decreased in the mutants. The triple mutant strain had the most powerful ability to produce optically pure D-lactate from glucose, and was tested with xylose and arabinose as carbon sources. Fed-batch fermentation was also carried out under various aeration rates, and the strain accumulated 125.1 g/L D-lactate with a yield of 0.91 g/g glucose at 2.5 vvm. Conclusions: Knocking out by-product synthesis genes had a remarkable influence on the production and yield of D-lactate. This study demonstrated, for the first time, that K. pneumoniae has great potential to convert monosaccharides into D-lactate. The results provide new insights for industrial production of D-lactate by K. pneumoniae. [ABSTRACT FROM AUTHOR]

Direct interspecies electron transfer (DIET) is an alternative to syntrophic metabolism in natural carbon cycle as well as in anaerobic digesters, but its function in anaerobic treatment of sulfate-containing wastewater have not yet to be described. Here, conductive stainless steel was added into anaerobic digesters for treating sulfate-containing wastewater to investigate the potential role of DIET in the response to the sulfate impact. Results showed that adding the conductive stainless steel made the anaerobic digestion less affected by the sulfate reduction than adding insulative plastic material. With adding stainless steel, methane production of the digesters increased by 7.5%-24.6%. Microbial analysis showed that the dissimilatory Fe (III) reducers like Geobacter species were enriched on the surface of the stainless steel. These results implied that the potential DIET of methanogenesis was established associating with stainless steel to outcompete the sulfate reduction. [ABSTRACT FROM AUTHOR]

Ewing sarcoma (EWS) is an aggressive bone malignancy that mainly affects children and young adults. The mechanisms by which EWS (EWSR1) fusion genes drive the disease are not fully understood. CRM1 (XPO1) traffics proteins from the nucleus, including tumor suppressors and growth factors, and is over-expressed in many cancers. A small-molecule inhibitor of CRM1, KPT-330, has shown therapeutic promise, but has yet to be investigated in the context of EWS. In this study, we demonstrate that CRM1 is also highly expressed in EWS. shRNA-mediated or pharmacologic inhibition of CRM1 in EWS cells dramatically decreased cell growth while inducing apoptosis, cell-cycle arrest, and protein expression alterations to several cancer-related factors. Interestingly, silencing of CRM1 markedly reduced EWS- FLI1 fusion protein expression at the posttranscriptional level and upregulated the expression of the well-established EWS-FLI1 target gene, insulin-like growth factor binding protein 3 (IGFBP3), which inhibits IGF-1. Accordingly, KPT-330 treatment attenuated IGF-1-induced activation of the IGF-1R/AKT pathway. Furthermore, knockdown of IGFBP3 increased cell growth and rescued the inhibitory effects on IGF-1 signaling triggered by CRM1 inhibition. Finally, treatment of EWS cells with a combination of KPT-330 and the IGF-1R inhibitor, linsitinib, synergistically decreased cell proliferation both in vitro and in vivo. Taken together, these findings provide a strong rationale for investigating the efficacy of combinatorial inhibition of CRM1 and IGF-1R for the treatment of EWS. Cancer Res; 76(9); 2687-97. [ABSTRACT FROM AUTHOR]

Background: Marine mud is an abundant and largely unexplored source of enzymes with unique properties that may be useful for industrial and biotechnological purposes. However, since most microbes cannot be cultured in the laboratory, a cultivation-independent metagenomic approach would be advantageous for the identification of novel enzymes. Therefore, with the objective of screening novel lipolytic enzymes, a metagenomic library was constructed using the total genomic DNA extracted from marine mud. Results: Based on functional heterologous expression, 34 clones that showed lipolytic activity were isolated. The five clones with the largest halos were identified, and the corresponding genes were successfully overexpressed in Escherichia coli. Molecular analysis revealed that these encoded proteins showed 48-79 % similarity with other proteins in the GenBank database. Multiple sequence alignment and phylogenetic tree analysis classified these five protein sequences as new members of known families of bacterial lipolytic enzymes. Among them, EST4, which has 316 amino acids with a predicted molecular weight of 33.8 kDa, was further studied in detail due to its strong hydrolytic activity. Characterization of EST4 indicated that it is an alkaline esterase that exhibits highest hydrolytic activity towards p-nitrophenyl butyrate (specific activity: 1389 U mg-1) at 45 °C and pH 8.0. The half-life of EST4 is 55 and 46 h at 40 and 45 °C, respectively, indicating a relatively high thermostability. EST4 also showed remarkable stability in organic solvents, retaining 90 % of its initial activity when incubated for 12 h in the presence of hydrophobic alkanes. Furthermore, EST4 was used as an efficient whole-cell biocatalyst for the synthesis of short-chain flavor esters, showing high conversion rate and good tolerance for high substrate concentrations (up to 3.0 M). These results demonstrate a promising potential for industrial scaling-up to produce short-chain flavor esters at high substrate concentrations in non-aqueous media. Conclusions: This manuscript reports unprecedented alcohol tolerance and conversion of an esterase biocatalyst identified from a marine mud metagenomic library. The high organic solvent tolerance and thermostability of EST4 suggest that it has great potential as a biocatalyst. [ABSTRACT FROM AUTHOR]

Background: Acute exacerbation is a common cause of hospitalization in patients with chronic heart failure, and coronary heart disease is the most common cause. Shenfu injection, a Traditional Chinese Medicine injection, widely used in the adjuvant treatment of patients with acute exacerbation of chronic heart failure, shows some treatment effect in improving the symptoms and the quality of life, but it lacks the rigorous clinical evaluation of research reports. This paper describes the protocol for the clinical assessment of Shenfu injection loading in the treatment of patients with acute exacerbation of chronic heart failure. Methods: This protocol adopts the design of a prospective, randomized, multicenter, blind imitation, placebo-controlled trial to assess the efficacy and safety of Shenfu injection loading in the treatment of patients with acute exacerbation of chronic heart failure due to coronary heart disease. The research will be carried out in 12 hospitals in China and is expected to enroll 160 inpatients with acute exacerbation of chronic heart failure due to coronary heart disease (yang and qi deficiency syndrome). On the basis of the conventional therapy of western medicine, patients will be randomized to either the treatment group (100 ml 5% glucose injection + 50 ml Shenfu injection) or the control group (150 ml 5% glucose injection) for 7 ± 1 days and follow-up for 28 ± 3 days. The primary outcomes are New York Heart Association cardiac function classification and Traditional Chinese Medicine syndromes. The secondary outcomes are left ventricular ejection fraction, brain natriuretic peptide level, Lee's heart failure score, 6-minute walking distance, and the incidence and readmission rate of cardiovascular events (including the emergency rate due to acute exacerbation of chronic heart failure). Discussion: This trial will assess the effect of loading Shenfu injection in the treatment of patients with acute exacerbation of chronic heart failure caused by coronary heart disease (yang-qi deficiency syndrome) on the symptoms and signs of heart failure, exercise tolerance, and other aspects, and observe its influence on the short-term prognosis with follow-up. The results of the study will provide clinical research evidence for application of Shenfu injection in the treatment. Trial registration: This trial was registered on 26 December 2012 at the Chinese Clinical Trials Register (Identifier: ChiCTR-TRC-12002857). [ABSTRACT FROM AUTHOR]

Osteosarcoma is a common malignant bone tumor with a propensityto metastasizetothe lungs. Epigenetic abnormalities have been demonstrated to underlie osteosarcoma development; however, the epigenetic mechanisms that are involved in metastasis are not yet clear. Here, we analyzed 2 syngeneic primary human osteosarcoma cell lines that exhibit disparate metastatic potential for differences in epigenetic modifications and expression. Using methylated DNA immunoprecipitation (MeDIP) and microarray expression analysis to screen for metastasis-associated genes, we identified Iroquois homeobox 1 [IRX1). In both human osteosarcoma cell lines and clinical osteosarcoma tissues, IRX1 overexpression was strongly associated with hypomethylation of its own promoter. Furthermore, experimental modulation of IRX1 in osteosarcoma cell lines profoundly altered metastatic activity, including migration, invasion, and resistance to anoikis in vitro, and influenced lung metastasis in murine models. These prometastatic effects of IRX1 were mediated by upregulation of CXCL14/NF-κB signaling. In serum from osteosarcoma patients, the presence of IRX1 hypomethylation in circulating tumor DNA reduced lung metastasis-free survival. Together, these results identify IRX1 as a prometastatic gene, implicate IRX1 hypomethylation as a potential molecular marker for lung metastasis, and suggest that epigenetic reversion of IRX1 activation may be beneficial for controlling osteosarcoma metastasis. [ABSTRACT FROM AUTHOR]

The mechanisms by which cancer cells migrate to metastasise are not fully understood. Breast cancers are accompanied by electrical depolarisation of tumour epithelial cells. The electrical changes can be detected on the skin and are used to differentiate malignant from benign breast tumours. Could the electrical signals play a role in metastasis by promoting tumour cell migration? We report that electric fields stimulate and direct migration of human breast cancer cells. Importantly, these effects were more significant in highly metastatic tumour cells than in low metastatic tumour cells. Electric-field-enhanced directional migration correlates well with the expression level of EGF receptor (EGFR/ErbB1). To confirm this, we transfected low metastatic clone MTC cells with human ErbB1, which significantly increased the electrotactic response. Inhibition of ErbB1 completely abolished the directional response of MTLn3 cells to an electric field. Transfection of MTLn3 cells and MDA-MB-435 cells with expression vectors for ErbB family members ErbB1, ErbB2 and ErbB3 also significantly enhanced EF-induced migration. These results suggest that electric signals might play a role in metastasis of breast cancers by enhancing cell migration through the ErbB-signalling pathway. [ABSTRACT FROM AUTHOR]