Identification and validation of genes with expression patterns inverse to multiple metastasis suppressor genes in breast cancer cell lines

Metastasis suppressor genes (MSGs) have contributed to an understanding of regulatory pathways unique to the lethal metastatic process. When re-expressed in experimental models, MSGs block cancer spread to, and colonization of distant sites without affecting primary tumor formation. Genes have been identified with expression patterns inverse to a single MSG, and found to encode functional, druggable signaling pathways. We now hypothesize that common signaling pathways mediate the effects of multiple MSGs. By gene expression profiling of human MCF7 breast carcinoma cells expressing a scrambled siRNA, or siRNAs to each of 19 validated MSGs (NME1, BRMS1, CD82, CDH1, CDH2, CDH11, CASP8, MAP2K4, MAP2K6, MAP2K7, MAPK14, GSN, ARHGDIB, AKAP12, DRG1, CD44, PEBP1, RRM1, KISS1), we identified genes whose expression was significantly opposite to at least five MSGs. Five genes were selected for further analysis: PDE5A, UGT1A, IL11RA, DNM3 and OAS1. After stable downregulation of each candidate gene in the aggressive human breast cancer cell line MDA-MB-231T, in vitro motility was significantly inhibited. Two stable clones downregulating PDE5A (phosphodiesterase 5A), an enzyme involved in the regulation of cGMP-specific signaling, exhibited no difference in cell proliferation, but reduced motility by 47 and 66 % compared to the empty vector-expressing cells (p = 0.01 and p = 0.005). In an experimental metastasis assay, two shPDE5A-MDA-MB-231T clones produced 47-62 % fewer lung metastases than shRNA-scramble expressing cells (p = 0.045 and p = 0.009 respectively). This study demonstrates that previously unrecognized genes are inversely related to the expression of multiple MSGs, contribute to aspects of metastasis, and may stand as novel therapeutic targets

A low-dissipation, pumpless, gravity-induced flow battery

Redox flow batteries have the potential to provide low-cost energy storage to enable renewable energy technologies such as wind and solar to overcome their inherent intermittency and to improve the efficiency of electric grids. Conventional flow batteries are complex electromechanical systems designed to simultaneously control flow of redox active fluids and perform electrochemical functions. With the advent of redox active fluids with high capacity density, i.e., Faradaic capacity significantly exceeding the 1–2 M concentration equivalents typical of aqueous redox flow batteries, new flow battery designs become of interest. Here, we design and demonstrate a proof-of-concept prototype for a “gravity-induced flow cell” (GIFcell), representing one of a family of approaches to simpler, more robust, passively driven, lower-cost flow battery architectures. Such designs are particularly appropriate for semi-solid electrodes comprising suspensions of networked conductors and/or electroactive particles, due to their low energy dissipation during flow. Accordingly, we demonstrate the GIFcell using nonaqueous lithium polysulfide solutions containing a nanoscale carbon network in a half-flow-cell configuration and achieve round trip energy efficiency as high as 91%

Reforestation in southern China: revisiting soil N mineralization and nitrification after 8 years restoration

Nitrogen availability and tree species selection play important roles in reforestation. However, long-term field studies on the effects and mechanisms of tree species composition on N transformation are very limited. Eight years after tree seedlings were planted in a field experiment, we revisited the site and tested how tree species composition affects the dynamics of N mineralization and nitrification. Both tree species composition and season significantly influenced the soil dissolved organic carbon (DOC) and nitrogen (DON). N-fixing Acacia crassicarpa monoculture had the highest DON, and 10-mixed species plantation had the highest DOC. The lowest DOC and DON concentrations were both observed in Eucalyptus urophylla monoculture. The tree species composition also significantly affected net N mineralization rates. The highest rate of net N mineralization was found in A. crassicarpa monoculture, which was over twice than that in Castanopsis hystrix monoculture. The annual net N mineralization rates of 10-mixed and 30-mixed plantations were similar as that of N-fixing monoculture. Since mixed plantations have good performance in increasing soil DOC, DON, N mineralization and plant biodiversity, we recommend that mixed species plantations should be used as a sustainable approach for the restoration of degraded land in southern China

Functional Characterization of Proanthocyanidin Pathway Enzymes from Tea and Their Application for Metabolic Engineering

Tea (Camellia sinensis) is rich in specialized metabolites, especially polyphenolic proanthocyanidins (PAs) and their precursors. To better understand the PA pathway in tea, we generated a complementary DNA library from leaf tissue of the blister blight-resistant tea cultivar TRI2043 and functionally characterized key enzymes responsible for the biosynthesis of PA precursors. Structural genes encoding enzymes involved in the general phenylpropanoid/flavonoid pathway and the PA-specific branch pathway were well represented in the library. Recombinant tea leucoanthocyanidin reductase (CsLAR) expressed in Escherichia coli was active with leucocyanidin as substrate to produce the 2R,3S-trans-flavan-ol (+)-catechin in vitro. Two genes encoding anthocyanidin reductase, CsANR1 and CsANR2, were also expressed in E. coli, and the recombinant proteins exhibited similar kinetic properties. Both converted cyanidin to a mixture of (+)-epicatechin and (−)-catechin, although in different proportions, indicating that both enzymes possess epimerase activity. These epimers were unexpected based on the belief that tea PAs are made from (−)-epicatechin and (+)-catechin. Ectopic expression of CsANR2 or CsLAR led to the accumulation of low levels of PA precursors and their conjugates in Medicago truncatula hairy roots and anthocyanin-overproducing tobacco (Nicotiana tabacum), but levels of oligomeric PAs were very low. Surprisingly, the expression of CsLAR in tobacco overproducing anthocyanin led to the accumulation of higher levels of epicatechin and its glucoside than of catechin, again highlighting the potential importance of epimerization in flavan-3-ol biosynthesis. These data provide a resource for understanding tea PA biosynthesis and tools for the bioengineering of flavanols

A WD40 Repeat Protein from Medicago truncatula

WD40 repeat proteins regulate biosynthesis of anthocyanins, proanthocyanidins (PAs), and mucilage in the seed and the development of trichomes and root hairs. We have cloned and characterized a WD40 repeat protein gene from Medicago truncatula (MtWD40-1) via a retrotransposon-tagging approach. Deficiency of MtWD40-1 expression blocks accumulation of mucilage and a range of phenolic compounds, including PAs, epicatechin, other flavonoids, and benzoic acids, in the seed, reduces epicatechin levels without corresponding effects on other flavonoids in flowers, reduces isoflavone levels in roots, but does not impair trichome or root hair development. MtWD40-1 is expressed constitutively, with highest expression in the seed coat, where its transcript profile temporally parallels those of PA biosynthetic genes. Transcript profile analysis revealed that many genes of flavonoid biosynthesis were down-regulated in a tissue-specific manner in M. truncatula lines harboring retrotransposon insertions in the MtWD40-1 gene. MtWD40-1 complemented the anthocyanin, PA, and trichome phenotypes of the Arabidopsis (Arabidopsis thaliana) transparent testa glabrous1 mutant. We discuss the function of MtWD40-1 in natural product formation in M. truncatula and the potential use of the gene for engineering PAs in the forage legume alfalfa (Medicago sativa)

Surface Morphology Evolution Mechanisms of InGaN/GaN Multiple Quantum Wells with Mixture N2/H2-Grown GaN Barrier

This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.Surface morphology evolution mechanisms of InGaN/GaN multiple quantum wells (MQWs) during GaN barrier growth with different hydrogen (H2) percentages have been systematically studied. Ga surface-diffusion rate, stress relaxation, and H2 etching effect are found to be the main affecting factors of the surface evolution. As the percentage of H2 increases from 0 to 6.25%, Ga surface-diffusion rate and the etch effect are gradually enhanced, which is beneficial to obtaining a smooth surface with low pits density. As the H2 proportion further increases, stress relaxation and H2 over- etching effect begin to be the dominant factors, which degrade surface quality. Furthermore, the effects of surface evolution on the interface and optical properties of InGaN/GaN MQWs are also profoundly discussed. The comprehensive study on the surface evolution mechanisms herein provides both technical and theoretical support for the fabrication of high-quality InGaN/GaN heterostructures.Peer reviewe

A pilot study monitoring the thermal comfort of the elderly living in nursing homes in Hefei, China, using wireless sensor networks, site measurements and a survey

Due to the rapid growth of the ageing population and the shrinking of family size in China, there has been a steady increase in the demand for nursing homes. The Chinese government has encouraged the development of nursing homes. However, there is no clear understanding of the thermal environment of nursing homes and the thermal comfort of their residents. Therefore, this paper describes a pilot study that was undertaken to investigate the thermal comfort of senior residents living in nursing homes. A wireless sensor network system was developed to continuously monitor the thermal environment of nursing homes for an entire year. Site measurements and a questionnaire also were conducted. The results revealed significant seasonal variations in the thermal environment of nursing homes and in the thermal comfort, thermal sensation and adaptive behaviours of older adults. The calculation of thermal comfort was modified according to the older adults’ behaviour patterns. The knowledge acquired in this initial study will enrich knowledge of the thermal comfort of older adults living in nursing homes. It also will provide preliminary insights for caregivers, designers and policymakers in their efforts to deliver a more comfortable, healthy and sustainable environment in nursing homes in China

Nm23-h1 Binds to Gelsolin and Inactivates Its Actin-Severing Capacity to Promote Tumor Cell Motility and Metastasis

Nm23-H1 has been identified as a metastasis suppressor gene, but its protein interactions have yet to be understood with any mechanistic clarity. In this study, we evaluated the proteomic spectrum of interactions made by Nm23-H1 in 4T1 murine breast cancer cells derived from tissue culture, primary mammary tumors, and pulmonary metastases. By this approach, we identified the actin-severing protein Gelsolin as binding partner for Nm23-H1, verifying their interaction by coimmunoprecipitation in 4T1 cells as well as in human MCF7, MDA-MB-231T, and MDA-MB-435 breast cancer cells. In Gelsolin-transfected cells, coexpression of Nm23-H1 abrogated the actin-severing activity of Gelsolin. Conversely, actin severing by Gelsolin was abrogated by RNA interference-mediated silencing of endogenous Nm23-H1. Tumor cell motility was negatively affected in parallel with Gelsolin activity, suggesting that Nm23-H1 binding inactivated the actin-depolymerizing function of Gelsolin to inhibit cell motility. Using indirect immunoflourescence to monitor complexes formed by Gelsolin and Nm23-H1 in living cells, we observed their colocalization in a perinuclear cytoplasmic compartment that was associated with the presence of disrupted actin stress fibers. In vivo analyses revealed that Gelsolin overexpression increased the metastasis of orthotopically implanted 4T1 or tail vein-injected MDA-MB-231T cells (P = 0.001 and 0.04, respectively), along with the proportion of mice with diffuse liver metastases, an effect ablated by coexpression of Nm23-H1. We observed no variation in proliferation among lung metastases. Our findings suggest a new actin-based mechanism that can suppress tumor metastasis